BR112017008568B1 - IMMUNORREGULATORY AGENTS - Google Patents

Abstract

IMMUNORREGULATORY AGENTS. The present invention relates to compounds that modulate the oxidoreductase enzyme, indoleamine 2,3-dioxygenase, and compositions containing the compounds are described herein. The use of such compounds and compositions for the treatment and/or prevention of a diverse range of diseases, disorders and conditions, including cancer and immune-related disorders, which are mediated by indoleamine 2,3-dioxygenase is also provided.

Description

CROSS REFERENCE TO RELATED ORDERS

[001] This application claims priority to United States Provisional Applications Serial Nos. 62/075,671, filed on November 5, 2014, and 62/098,022, filed on December 30, 2014, respectively, the full content of which is incorporated herein by reference.

BACKGROUND OF THE INVENTION

[002] Indoleamine 2,3-dioxygenase (IDO; also known as IDO1) is an IFN-Y target gene that plays a role in immunomodulation. IDO is an oxidoreductase and one of two enzymes that catalyze the first and rate-limiting step in the conversion of tryptophan to N-formyl-kynurenine. It exists as a 41kD monomer that is found in diverse cell populations, including immune cells, endothelial cells, and fibroblasts. IDO is relatively well conserved across species, with mouse and human sharing 63% sequence identity at the amino acid level. Data derived from its crystal structure and site-directed mutagenesis show that both substrate binding and the relationship between substrate and iron-bound dioxygenase are necessary for activity. A homologue of IDO (IDO2) has been identified that shares 44% amino acid sequence homology with IDO, but its function is largely distinct from that of IDO. (See, for example, Serafini P, et al., Semin. Cancer Biol., 16(1):53-65 (February 2006) and Ball, H.J. et al., Gene, 396(1):203-213 (July 2007)).

[003] IDO plays an important role in immunoregulation, and its immunosuppressive function manifests itself in several ways. Importantly, IDO regulates immunity at the T cell level, and a nexus exists between IDO and cytokine production. Furthermore, tumors often manipulate function by upregulating IDO. Thus, IDO modulation can have a therapeutic impact on various diseases, disorders and conditions.

[004] A pathophysiological link exists between IDO and cancer. Disruption of immunological homeostasis is intimately involved with tumor growth and progression, and IDO production in the tumor microenvironment appears to aid tumor growth and metastasis. Furthermore, increased levels of IDO activity are associated with a variety of different tumors (Brandacher, G. et al., Clin. Cancer Res., 12(4):1144-1151 (February 15, 2006)).

[005] Cancer treatment commonly involves surgical resection followed by chemotherapy and radiotherapy. Standard treatment regimens show varying degrees of long-term success because of the ability of tumor cells to essentially escape by regeneration of primary tumor growth and, often more importantly, seeding of distant metastasis. Recent advances in the treatment of cancer and cancer-related diseases, disorders and conditions comprise the use of combination therapy incorporating immunotherapy with more traditional chemotherapy and radiotherapy. In most scenarios, immunotherapy is associated with less toxicity than traditional chemotherapy because it uses the patient's own immune system to identify and eliminate tumor cells.

[006] In addition to cancer, IDO has been implicated in, among other conditions, immunosuppression, chronic infections, and autoimmune diseases or disorders (e.g., rheumatoid arthritis). Therefore, the suppression of tryptophan degradation by inhibition of IDO activity has enormous therapeutic value. Additionally, IDO inhibitors can be used to enhance T cell activation when T cells are suppressed by pregnancy, malignancy, or a virus (e.g., HIV). Although their roles are not well defined, IDO inhibitors may also find use in treating patients with neurological or neuropsychiatric diseases or disorders (e.g., depression).

[007] Small molecule IDO inhibitors have been developed to treat or prevent IDO-related diseases. For example, the IDO inhibitors, 1-methyl-DL-tryptophan; p-(3-benzofuranyl)-DL-alanine; p-[3-benzo(b)thienyl]-DL-alanine; and 6-nitro-L-tryptophan have been used to modulate T cell-mediated immunity by altering local extracellular concentrations of tryptophan and tryptophan metabolites (WO 99/29310). Compounds having IDO inhibitory activity are also reported in WO 2004/094409.

[008] In view of the role played by indoleamine 2,3-dioxygenase in a diverse range of diseases, disorders and conditions, and the limitations (e.g., efficacy) of current IDO inhibitors, new IDO modulators, and compositions and methods associated with these, are necessary.

BRIEF SUMMARY OF THE INVENTION

[009] The present invention relates to compounds that modulate the oxidoreductase enzyme, indoleamine 2,3-dioxygenase (IDO), and compositions (for example, pharmaceutical compositions) comprising the compounds. Such compounds, including methods of their synthesis, and compositions are described in detail below.

[0010] The present invention also relates to the use of such compounds and compositions for the treatment and/or prevention of a diverse series of diseases, disorders and conditions mediated, in whole or in part, by IDO. Such diseases, disorders and conditions are described in detail elsewhere herein. Unless otherwise indicated, when uses of the compounds of the present invention are described herein, it is to be understood that such compounds may be in the form of a composition (e.g., a pharmaceutical composition).

[0011] As described below, although the compounds of the present invention are believed to achieve their activity by inhibiting IDO, a precise understanding of the underlying mechanism of action of the compounds is not required to practice the invention. It is considered that the compounds may alternatively achieve their activity through inhibition of tryptophan-2,3-dioxygenase (TDO) activity. It is also considered that the compounds can achieve their activity through inhibition of both IDO and TDO function. Although the compounds of the invention are generally referred to herein as IDO inhibitors, it is to be understood that the term "IDO inhibitors" encompasses compounds that act individually through inhibition of either TDO or IDO, and/or compounds that act through inhibition of both IDO as TDO.

[0012] In one aspect, the present invention provides compounds represented by formula (I): or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein, the subscript n is 1 or 0; the subscript p is 1 or 0; the ring designated as A is phenyl, 5- or 6-membered heteroaryl, or C5-7 cycloalkyl; Z is O; B is N, C(OR5a), or C(R3a); each X is independently NR5a, O, CHR5, C(O), or CH(OR5a); Q is N, C(CN), or CR6; D is a bond, O, C(R5)2, or NR5a; E is an optionally substituted 9- or 8-membered fused bicyclic heteroaryl; R1 and R2 are independently hydrogen, halogen, C1-C4 haloalkyl, C3-C6 cycloalkyl, 3- to 6-membered cycloheteroalkyl, optionally substituted phenyl, optionally substituted heteroaryl, C1-C4- optionally substituted alkyl, C1-C4 alkoxy, CN , SO2NH2, NHSO2CH3, NHSO2CF3, OCF3, SO2CH3, SO2CF3, or CONH2, and when R1 and R2 are at adjacent vertices of a phenyl ring they can be linked together to form a 5- or 6-membered cycloheteroalkyl ring having one or two ring vertices independently selected from O, N and S, wherein said cycloheteroalkyl ring is optionally substituted with one to three members selected from fluorine and C1-C3 alkyl; R3, R3a and R4 are independently hydrogen, C1-C6 optionally substituted alkyl, C2-C6 optionally substituted alkenyl, C2-C6 optionally substituted alkynyl, C1-C6 optionally substituted haloalkyl, C3-C6 cycloalkyl-C1-C4 optionally substituted alkyl, aryl -C1-C6 optionally substituted alkyl, fluorine, OH, CN, CO2H, C(O)NH2, N(R5)2, -O-C1-C6 optionally substituted alkyl, -(CR5R5)m-OH, - (CR5R5) m-CO2H, -(CR5R5)mC(O)NH2, -(CR5R5)mC(O)NHR5, - (CR5R5)mN(R5)2, -NH(CR5R5)mCO2H or -NH(CR5R5)mC(O) NH2; each R5 is independently H, F, OH, or optionally substituted C1-C6 alkyl; each R5a is independently H, or optionally substituted C1-C6 alkyl; R6 is H, OH, F, optionally substituted C1-C6 alkyl, -O-C1-C6 optionally substituted alkyl, or -N(R5a)2; and each m is independently 1, 2, or 3.

[0013] In yet another aspect, the present invention provides compositions in which compounds of formula (I) are combined with one or more pharmaceutically acceptable excipients.

[0014] In some embodiments, the present invention contemplates methods for treating or preventing cancer in an individual (e.g., a human) comprising administering to the individual a therapeutically effective amount of at least one IDO inhibitor described herein. The present invention includes methods of treating or preventing a cancer in a subject by administering to the subject an IDO inhibitor in an amount effective to reverse or halt the progression of IDO-mediated immunosuppression. In some embodiments, IDO-mediated immunosuppression is mediated by an antigen presenting cell (APC).

[0015] Examples of cancers that can be treated using the compounds and compositions described herein include, but are not limited to: cancers of the prostate, colorectal, pancreas, cervix, stomach, endometrium, brain, liver, bladder, ovary, testis, head , neck, skin (including melanoma and basal cell carcinoma), mesothelial lining, white blood cells (including lymphoma and leukemia), esophagus, breast, muscle, connective tissue, lung (including small cell carcinoma and non-small cell carcinoma), adrenal gland, thyroid, kidney, or bone; glioblastoma, mesothelioma, renal cell carcinoma, gastric carcinoma, sarcoma, choriocarcinoma, cutaneous basal cell carcinoma, and testicular seminoma. In some embodiments of the present invention, the cancer is melanoma, colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, leukemia, a brain tumor, lymphoma, sarcoma, ovarian cancer, or Kaposi's sarcoma. Cancers that are candidates for treatment with the compounds and compositions of the present invention are also described hereinafter.

[0016] The present invention contemplates methods of treating an individual receiving a bone marrow transplant or peripheral blood stem cell transplant by administering a therapeutically effective amount of an IDO inhibitor sufficient to increase the delayed-type hypersensitivity reaction to the IDO antigen. tumor, delay time-to-recurrence of malignancy after transplantation, increase recurrence-free survival time, and/or increase long-term survival after transplantation.

[0017] In certain embodiments, the present invention contemplates methods for treating or preventing an infectious disorder (e.g., a viral infection) in an individual (e.g., a human) comprising administering to the individual a therapeutically effective amount of hair at least one IDO inhibitor (e.g., a new inhibitor of the present invention). In some embodiments, the infectious disorder is a viral infection (e.g., a chronic viral infection), a bacterial infection, or a parasitic infection. In certain embodiments, the viral infection is human immunodeficiency virus or cytomegalovirus. In other embodiments, the bacterial infection is an infection by Mycobacteria (e.g., Mycobacterium leprae or Mycobacterium tuberculosis). In still other embodiments, the parasitic infection is Leishmania donovani, Leishmania tropica, Leishmania major, Leishmania aethiopica, Leishmania Mexicana, Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale, or Plasmodium malariae. In other embodiments, the infectious disorder is a fungal infection.

[0018] In still other embodiments, the present invention contemplates methods for treating or preventing a disease, disorder or immune-related condition in an individual (e.g., a human), comprising administering to the individual a therapeutically effective amount of at least one inhibitor of IDO (e.g., preferably a new inhibitor of the present invention). Examples of immune-related diseases, disorders and conditions are described below.

[0019] Other diseases, disorders and conditions that can be treated or prevented, in whole or in part, by modulating IDO activity are candidate indications for the IDO inhibitor compounds that are described here.

[0020] The present invention also contemplates the use of the IDO inhibitors described herein in combination with one or more additional agents. One or more additional agents may have some IDO modulating activity and/or they may function through distinct mechanisms of action. In some embodiments, such agents comprise radiation (e.g., localized radiation therapy or total body radiation therapy) and/or other treatment modalities of a non-pharmacological nature. When combination therapy is used, the IDO inhibitor(s) and the additional agent(s) may be in the form of a single composition or multiple compositions, and the treatment modalities may be administered concurrently, sequentially, or through some another treatment regimen. By way of example, the present invention contemplates a treatment regimen in which a radiation phase is followed by a chemotherapeutic phase. Combination therapy may have an additive or synergistic effect. Other benefits of combination therapy are described here below.

[0021] In some embodiments, the present invention also comprises the use of the IDO inhibitors described herein in combination with bone marrow transplantation, peripheral blood stem cell transplantation, or other types of transplantation therapy.

[0022] In particular embodiments, the present invention contemplates the use of the IDO function inhibitors described here in combination with immunological checkpoint inhibitors. Immune checkpoint blockade, which results in amplification of antigen-specific T cell responses, has been shown to be a promising method in human cancer therapeutics. Examples of immune checkpoints (ligands and receptors), some of which are selectively upregulated on various tumor cell types, that are candidates for blockade include PD1 (programmed cell death protein 1); PDL1 (PD1 ligand); BTLA (B and T lymphocyte attenuator); CTLA4 (cytotoxic T lymphocyte-associated antigen 4); TIM3 (T cell membrane protein 3); LAG3 (lymphocyte activation gene 3); A2aR (A2a adenosine receptor A2aR); and Killer inhibitory receptors. Immune checkpoint inhibitors, and combination therapy therewith, are described in detail elsewhere here.

[0023] In other embodiments, the present invention provides methods for treating cancer in an individual, comprising administering to the individual a therapeutically effective amount of at least one IDO inhibitor and at least one chemotherapeutic agent, such agents including, but not limited to alkylating agents (e.g., nitrogen mustards such as chlorambucil, cyclophosphamide, isofamide, mechlorethamine, melphalan, and uracil mustard; aziridines such as thiotepa; methanesulfonate esters such as busulfan; nucleoside analogs (e.g., gemcitabine); nitrous ureas such as carmustine, lomustine, and streptozocin; topoisomerase 1 inhibitors (e.g., irinotecan); platinum complexes such as cisplatin and carboplatin; bioreductive alkylators such as mitomycin, procarbazine, dacarbazine, and altretamine); DNA strand disrupting agents (e.g., bleomycin); topoisomerase II inhibitors (e.g., ansacrine, dactinomycin, daunorubicin, idarubicin, mitoxantrone, doxorubicin, etoposide, and teniposide); DNA minor groove binding agents (e.g., plicamidine); antimetabolites (e.g., folate antagonists such as methotrexate and trimetrexate; pyrimidine antagonists such as fluorouracil, fluorodeoxyuridine, CB3717, azacitidine, cytarabine, and floxuridine; purine antagonists such as mercaptopurine, 6-thioguanine, fludarabine, pentostatin ; asparginase; and ribonucleotide reductase inhibitors such as hydroxyurea); tubulin interacting agents (e.g., vincristine, estramustine, vinblastine, docetaxol, epothilone derivatives, and paclitaxel); hormonal agents (e.g., estrogens; conjugated estrogens; ethinyl estradiol; diethylstilbesterol; chlortrianisen; idenestrol; progestins such as hydroxyprogesterone caproate, medroxyprogesterone, and megestrol; and androgens such as testosterone, testosterone propionate, fluoxymesterone, and methyltestosterone); adrenal corticosteroids (e.g., prednisone, dexamethasone, methylprednisolone, and prednisolone); luteinizing hormone-releasing agents or gonadotropin-releasing hormone antagonists (e.g., leuprolide acetate and goserelin acetate); and antihormonal antigens (e.g., tamoxifen, antiandrogen agents such as flutamide; and antiadrenal agents such as mitotane and aminoglutethimide). The present invention also contemplates the use of the IDO inhibitors in combination with other agents known in the art (e.g., arsenic trioxide) and other chemotherapeutic agents developed in the future.

[0024] In some embodiments required for cancer treatment methods, administration of a therapeutically effective amount of an IDO inhibitor in combination with at least one chemotherapeutic agent results in a cancer survival rate greater than the rate of cancer survival observed administering either alone. In other embodiments described for cancer treatment methods, administration of a therapeutically effective amount of an IDO inhibitor in combination with at least one chemotherapeutic agent results in a reduction in tumor size or a decrease in tumor growth greater than the reduction in tumor size or tumor growth observed by administration of an agent alone.

[0025] In other embodiments, the present invention contemplates methods for treating or preventing cancer in an individual, comprising administering to the individual a therapeutically effective amount of at least one IDO inhibitor and at least one signal transduction inhibitor (STI). ). In a particular embodiment, at least one STI is selected from the group consisting of bcr/abl kinase inhibitors, epidermal growth factor (EGF) receptor inhibitors, her-2/neu receptor inhibitors, and farnesyl transferases (FTIs). Other candidate STI agents are mentioned elsewhere here.

[0026] The present invention also contemplates methods of increasing the rejection of tumor cells in an individual comprising administering an IDO inhibitor in conjunction with at least one chemotherapeutic agent and/or radiation therapy, wherein the resulting rejection of tumor cells is greater than that obtained by administering the IDO inhibitor, chemotherapeutic agent, or radiation therapy alone.

[0027] In other embodiments, the present invention provides methods for treating cancer in an individual, comprising administering to the individual a therapeutically effective amount of at least one IDO inhibitor and at least one immunomodulator other than an IDO inhibitor. In particular embodiments, at least one immunomodulator is selected from the group consisting of CD40L, B7, B7RP1, anti-CD40, anti-CD38, anti-ICOS, 4-IBB ligand, dendritic cell cancer vaccine, IL2, IL12 , ELC/CCL19, SLC/CCL21, MCP-1, IL-4, IL-18, TNF, IL-15, MDC, IFN-α/-β, M-CSF, IL-3, GM-CSF, IL- 13, and anti-IL-10. Other candidate immunomodulatory agents are mentioned elsewhere herein.

[0028] The present invention contemplates embodiments comprising methods for treating or preventing an infectious disorder (e.g., a viral infection) in an individual (e.g., a human) comprising administering to the individual a therapeutically effective amount of at least one inhibitor of IDO is a therapeutically effective amount of an anti-infective agent(s).

[0029] In some embodiments of the present invention, the additional therapeutic agent is a cytokine, including, for example, granulocyte-macrophage colony-stimulating factor (GM-CSF) or flt3-ligand. The present invention also contemplates methods for treating or preventing a viral infection (e.g., a chronic viral infection) including, but not limited to, hepatitis C virus (HCV), human papilloma virus (HPV), cytomegalovirus (CMV) , Epstein-Barr virus (EBV), varicella zoster virus, coxsackie virus, and human immunodeficiency virus (HIV). The use of the IDO inhibitors described herein to treat (alone or as a component of combination therapy) infection is also described hereinafter.

[0030] In additional embodiments, treatment of an infectious disorder is accomplished through co-administration of a vaccine in combination with administration of a therapeutically effective amount of an IDO inhibitor of the present invention. In some embodiments, the vaccine is an antiviral vaccine, including, for example, an anti-HIV vaccine. In other embodiments, the vaccine is effective against tuberculosis or malaria. In still other embodiments, the vaccine is a tumor vaccine (e.g., an effective melanoma vaccine); The tumor vaccine may comprise genetically modified tumor cells or a genetically modified cell line, including genetically modified tumor cells or a genetically modified cell line that has been transfected to express granulocyte-macrophage stimulating factor (GM-CSF). In particular embodiments, the vaccine includes one or more immunogenic peptides and/or dendritic cells.

[0031] In some embodiments, the present invention contemplates methods of using the IDO inhibitors described herein in combination with one or more antimicrobial agents.

[0032] In certain embodiments required for treating an infection by administering an IDO inhibitor and at least one additional therapeutic agent, an infection symptom observed after administration of both the IDO inhibitor and the additional therapeutic agent is improved compared to the same symptom of infection observed after administration either alone. In some embodiments, the symptom of infection is observed by reduced viral load, increased CD4+ T cell count, decreased opportunistic infections, increased survival time, eradication of chronic infection, or a combination thereof.

BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1A-1P provides a structure and biological activity for the compounds described here. Inhibitory potencies measured for the compounds of the invention in the assay described in Example 251 are provided below in Figure 1A - 1P, wherein the potency levels are given as follows: (IDO potency: IC50: A < 0.1 μM; B < 1 μM; C < 10 μM).

DETAILED DESCRIPTION OF THE INVENTION

[0034] Before the present invention is further described, it should be understood that the invention is not limited to the particular embodiments mentioned herein, and it should also be understood that the terminology used herein is for the purposes of describing the particular embodiments only, and It is not intended to be limiting.

[0035] Where a range of values is provided, it is understood that each intervening value, to the tenth of a unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other value provided or intervening in that established range, is covered within the invention. The upper and lower limits of these minor ranges may independently be included in the minor ranges, and are also encompassed within the invention, subject to any limit specifically excluded in the established range. Where the established range includes one or both limits, ranges including either or both of those included limits are also included in the invention. Unless otherwise defined, technical and scientific terms used herein have the same meaning as understood by one skilled in the art to which this invention belongs.

[0036] It should be noted that as used herein and in the appended claims, the singular forms "a, a(a)", "a, a(an)", and "the, a" include referent plurals, unless the context clearly dictates otherwise. It is also noted that claims may be drafted to exclude any optional elements. As such, this statement is understood to function as an antecedent basis for use of such exclusive terminology such as "exclusively", "only" and the like in connection with the citation of claim elements, or use of a "negative" limitation.

[0037] The publications described here are provided solely for your description prior to the filing date of this application. Furthermore, the publication dates provided may differ from the actual publication dates, which may necessarily be independently confirmed.

General

[0038] Immune dysregulation is closely associated with tumor evasion of the host's immune system, resulting in tumor growth and progression. Traditional treatment methods comprising chemotherapy and radiotherapy are often difficult for the patient to tolerate and become less effective as tumors develop to survive such treatments. Using the patient's own immune system to identify and eliminate tumor cells, immunotherapy has the benefit of reduced toxicity. Since upregulation of the immunoregulatory enzyme, indoleamine 2,3-dioxygenase, comprises a mechanism manipulated by tumors to promote growth, agents (e.g., small molecule compounds) that inhibit enzyme activity present a pathway promising for prophylaxis and/or treatment.

[0039] Furthermore, a large body of experimental data indicates a role for IDO inhibition in immunosuppression, tumor resistance and/or rejection, chronic infections, HIV infection, and diseases and disorders. IDO inhibition may also be an important treatment strategy for patients with neurological or neuropsychiatric diseases or disorders such as depression. The compounds, compositions, and methods herein address the need for new classes of IDO modulators.

Definitions

[0040] Unless otherwise indicated, the following terms are intended to have the meaning set forth below. Other terms are defined elsewhere throughout the specification.

[0041] The term "alkyl", alone or as part of another substituent, means, unless otherwise stated, a straight or branched chain hydrocarbon radical, having the designated number of carbon atoms (i.e. is, C1-8 means one to eight carbons). Examples of alkyl groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, t-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, n-heptyl, n-octyl, and the like. The term "alkenyl" refers to an unsaturated alkyl group having one or more double bonds. Similarly, the term "alkynyl" refers to an unsaturated alkyl group having one or more triple bonds. Examples of such unsaturated alkyl groups include vinyl, 2-propenyl, crotyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl, and higher isomers and homologs.

[0042] The term "cycloalkyl" refers to hydrocarbon rings having the indicated number of ring atoms (e.g., C3-6 cycloalkyl) and being fully saturated or having no more than one double bond between the apexes of ring. "Cycloalkyl" is also meant to refer to bicyclic and polycyclic hydrocarbon rings such as, for example, bicyclo[2,2,1]heptane, bicyclo[2,2.2]octane, etc.

[0043] The term "cycloheteroalkyl" refers to a cycloalkyl ring having the indicated number of ring vertices (or members) and having one to five heteroatoms selected from N, O, and S, which replace one to five of the vertices carbon, and in which the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atom(s) are optionally quaternized. The cycloheteroalkyl may be a monocyclic, bicyclic or polycyclic ring system. Non-limiting examples of cycloheteroalkyl groups include pyrrolidine, imidazolidine, pyrazolidine, butyrolactam, valerolactam, imidazolidinone, hydantoin, dioxolane, phthalimide, piperidine, 1,4-dioxane, morpholine, thiomorpholine, thiomorpholine-S-oxide, thiomorpholine -S,S-oxide, piperazine, pyran, pyridone, 3-pyrroline, thiopyran, pyrone, tetrahydrofuran, tetrahydrothiophene, quinuclidine, and the like. A cycloheteroalkyl group can be linked to the rest of the molecule through a ring carbon or a heteroatom.

[0044] As used herein, a wavy line, "—", which intersects a single, double or triple bond in any chemical structure described herein, represents the point of attachment of the single, double or triple bonds to the rest of the molecule. Additionally, a bond that extends to the center of a ring (e.g., a phenyl group) is intended to indicate bonding at any of the ring vertices. One skilled in the art will understand that multiple substituents shown to be attached to a ring will occupy ring vertices that provide stable compounds and are otherwise sterically compatible. For a divalent component, a representation is intended to include any orientation (forward or reverse). For example, the group "-C(O)NH-" is intended to include a bond in either orientation: -C(O)NH- or -NHC(O)-, and similarly, "-O- CH2CH2-" is intended to include both -O-CH2CH2- and -CH2CH2-O-.

[0045] The terms "alkoxy", "alkylamino" and "alkylthio" (or thioalkoxy) are used in their conventional sense, and refer to those alkyl groups linked to the rest of the molecule through an oxygen atom, an amino group , or a sulfur atom, respectively. Additionally, for dialkylamino groups, the alkyl moieties may be the same or different and may also be combined to form a 3- to 7-membered ring with the nitrogen atom to which each is attached. Accordingly, a group represented as dialkylamino or -NRaRb is intended to include piperidinyl, pyrrolidinyl, morpholinyl, azetidinyl and the like.

[0046] The terms "halo" or "halogen", by themselves or as part of another substituent, mean, unless otherwise stated, an atom of fluorine, chlorine, bromine, or iodine. Additionally, terms such as "haloalkyl" are intended to include monohaloalkyl and polyhaloalkyl. For example, the term "C 1-4 haloalkyl" is intended to include trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like.

[0047] The term "aryl" means, unless otherwise stated, a polyunsaturated, typically aromatic hydrocarbon group that may be a single ring or multiple rings (up to three rings) that are fused together or covalently linked. Non-limiting examples of aryl groups include phenyl, naphthyl and biphenyl.

[0048] The term "heteroaryl" refers to aryl groups (or rings) that contain one to five heteroatoms selected from N, O, and S, in which the nitrogen and sulfur atoms are optionally oxidized, and the nitrogen atoms nitrogen are optionally quaternized. A heteroaryl group can be linked to the rest of the molecule through a heteroatom. Non-limiting examples of heteroaryl groups include pyridyl, pyridazinyl, pyrazinyl, pyrimidinyl, triazinyl, quinolinyl, quinoxalinyl, quinazolinyl, cinnolinyl, phthalazinyl, benzotriazinyl, purinyl, benzimidazolyl, benzopyrazolyl, benzotriazolyl, benzisoxazolyl, isobenzofuryl, isoindolyl, indolizinyl , benzotriazinyl, thienopyridinyl , thienopyrimidinyl, pyrazolopyrimidinyl, imidazopyridines, benzothiaxolyl, benzofuranyl, benzothienyl, indolyl, quinolyl, isoquinolyl, isothiazolyl, pyrazolyl, indazolyl, pteridinyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiadiazolyl, pyrrolyl, thiazolyl, furyl , thienyl and similar . Substituents for a heteroaryl ring may be selected from the group of acceptable substituents described below.

[0049] The terms above (for example, "alkyl", "aryl" and "heteroaryl"), in some embodiments, will be optionally replaced. Selected substituents for each type of radical are given below.

[0050] Optional substituents for alkyl radicals (including those groups often referred to as alkylene, alkenyl, alkynyl and cycloalkyl) may be a variety of groups selected from: halogen, -OR', -NR'R'', -SR', -SiR'R"R"', -OC(O)R', - C(O)R', -CO2R', -CONR'R'', -OC(O)NR'R'', -NR' 'C(O)R', -NR'- C(O)NR''R''', -NR''C(O)2R', -NH-C(NH2)=NH, -NR'C( NH2)=NH, -NH- C(NH2)=NR', -S(O)R', -S(O)2R', -S(O)2NR'R'', -NR'S(O)2R' ', -CN and -NO2 in a number ranging from zero to (2 m'+1), where m' is the total number of carbon atoms in such a radical. R', R'' and R''' each independently refers to hydrogen, C1-8 unsubstituted alkyl, unsubstituted aryl, aryl substituted with 1-3 halogen groups, C1-8 unsubstituted alkyl, C1-8 alkoxy or C1-8 thioalkoxy, or unsubstituted aryl-C1-4 alkyl groups. When R' and R'' are bonded to the same nitrogen atom, they can be combined with the nitrogen atom to form a 3-, 4-, 5-, 6-, or 7-membered ring. For example, -NR'R" is intended to include 1-pyrrolidinyl and 4-morpholinyl.

[0051] Similarly, optional substituents for the aryl and heteroaryl groups are varied and are generally selected from: - halogen, -OR', -OC(O)R', -NR'R", -SR', -R', -CN, -NO2, -CO2R', - CONR'R'', -C(O)R', -OC(O)NR'R'', -NR''C(O)R', -NR' 'C(O)2R', -NR'- C(O)NR''R''', -NH-C(NH2)=NH, -NR'C(NH2)=NH, -NH-C(NH2 )=NR', - S(O)R', -S(O)2R', -S(O)2NR'R'', -NR'S(O)2R'', -N3, perfluoro(C1- C4) alkoxy, and perfluoro(C1-C4)alkyl, in a number ranging from zero to the total number of open valences in the aromatic ring system; and where R', R'' and R''' are independently selected from hydrogen, C1- 8 alkyl, C1-8 haloalkyl, C3-6 cycloalkyl, C2-8 alkenyl and C2-8 alkynyl. Other suitable substituents include each of the above aryl substituents bonded to a ring atom by an alkylene ether of 1 to 4 atoms of carbon.

[0052] Two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be replaced with a substituent of the formula -T-C(O)-(CH2)q-U-, where T and U are independently -NH-, -O -, -CH2- or a single bond, and q is an integer from 0 to 2. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be substituted with a substituent of the formula -A-(CH2)r-B- , where A and B are independently -CH2-, -O-, -NH-, -S-, -S(O)-, -S(O)2-, -S(O)2NR'- or a bond simple, and r is an integer from 1 to 3. One of the single bonds of the new ring thus formed can optionally be replaced with a double bond. Alternatively, two of the substituents on adjacent atoms of the aryl or heteroaryl ring may optionally be substituted with a substituent of the formula -(CH2)s-X- (CH2)t-, where s and t are independently integers from 0 to 3, and X is -O-, -NR'-, -S-, -S(O)-, -S(O)2-, or -S(O)2NR'-. The substituent R' in -NR'- and -S(O)2NR'- is selected from hydrogen or unsubstituted C1-6 alkyl.

[0053] As used herein, the term "heteroatom" is intended to include oxygen (O), nitrogen (N), sulfur (S) and silicon (Si).

[0054] The term "pharmaceutically acceptable salts" is intended to include salts of active compounds that are prepared with relatively non-toxic acids or bases, depending on the particular substituents found in the compounds described here. When the compounds of the present invention contain relatively acidic functionalities, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either pure or in a suitable inert solvent. Examples of salts derived from pharmaceutically acceptable inorganic bases include aluminum, ammonium, calcium, copper, ferric, ferrous, lithium, magnesium, manganic, manganous, potassium, sodium, zinc and the like. Salts derived from pharmaceutically acceptable organic bases include salts of primary, secondary and tertiary amines, including substituted amines, cyclic amines, naturally occurring amines and the like, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2 -diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine , triethylamine, trimethylamine, tripropylamine, tromethamine and the like. When the compounds of the present invention contain relatively basic functionalities, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired acid, either alone or in a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include those derived from inorganic acids such as hydrochloric, hydrobromic, nitric, carbonic, monohydrocarbonic, phosphoric, monohydrogenphosphoric, dihydrogenphosphoric, sulfuric, monohydrogensulfuric, hydrodic, or phosphorous acids and the like, as well as salts derived from relatively non-toxic organic acids such as acetic, propionic, isobutyric, malonic, benzoic, succinic, suberic, fumaric, mandelic, phthalic, benzenesulfonic, p-tolylsulfonic, citric, tartaric, methanesulfonic, and the like . Also included are salts of amino acids such as arginate and the like, and salts of organic acids such as glucuronic or galactunoric acids and the like (see, for example, Berge, S.M., et al., "Pharmaceutical Salts", J. Pharm. Sci. , 66:1-19 (1977)). Certain specific compounds of the present invention contain both basic and acidic functionalities that allow the compounds to be converted into base or acid addition salts.

[0055] The neutral forms of the compounds can be regenerated by contacting the salt with a base or acid and isolating the original compound in the conventional manner. The parent form of the compound differs from the various salt forms in certain physical properties, such as solubility in polar solvents, but otherwise the salts are equivalent to the parent form of the compound for the purposes of the present invention.

[0056] In addition to salt forms, the present invention provides compounds that are in a prodrug form. The prodrugs of the compounds described herein are those compounds that readily undergo chemical changes under physiological conditions to provide the compounds of the present invention. Additionally, prodrugs can be converted into the compounds of the present invention by chemical or biochemical methods in an ex vivo environment. For example, prodrugs can be slowly converted into the compounds of the present invention when placed in a transdermal patch reservoir with a suitable enzyme or chemical reagent.

[0057] Certain compounds of the present invention can exist in unsolvated forms as well as solvated forms, including hydrated forms. In general, solvated forms are equivalent to unsolvated forms and are intended to be encompassed within the scope of the present invention. Certain compounds of the present invention may exist in multiple crystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.

[0058] Certain compounds of the present invention have asymmetric carbon atoms (optical centers) or double bonds; Racemates, diastereomers, geometric isomers, regioisomers and individual isomers (e.g., separate enantiomers) are all intended to be encompassed within the scope of the present invention. When a stereochemical description is shown, it is intended to refer to the compound in which one of the isomers is present and substantially free of the other isomer. "Substantially free from" another isomer indicates at least an 80/20 ratio of the two isomers, more preferably 90/10, or 95/5 or more. In some embodiments, one of the isomers will be present in an amount of at least 99%.

[0059] The compounds of the present invention may also contain unnatural proportions of atomic isotopes of one or more of the atoms that constitute such compounds. Unnatural proportions of an isotope can be defined as ranging from the amount found in nature to an amount ranging from 100% of the atom in question. For example, the compounds may incorporate radioactive isotopes, such as, for example, tritium (3H), iodine-125 (125I), or carbon-14 (14C), or non-radioactive isotopes, such as deuterium (2H) or carbon. -13 (13C). Such isotopic variations may provide additional utilities to those elsewhere in this application. For example, isotopic variants of the compounds of the invention may find additional utility, including, but not limited to, as diagnostic and/or imaging reagents, or as cytotoxic/radiotoxic therapeutic agents. Additionally, isotopic variants of compounds may have pharmacokinetic and pharmacodynamic characteristics that may contribute to safety, tolerability or enhanced efficacy during treatment. All variations of the compounds of the present invention, whether radioactive or not, are intended to be encompassed within the scope of the present invention.

[0060] The terms "patient" or "individual" are used interchangeably to refer to a human or a non-human animal (e.g., a mammal).

[0061] The terms "administration", "administer" and the like, when applying to, for example, an individual, cell, tissue, organ, or biological fluid, refer to the contact of, for example, an IDO inhibitor , a pharmaceutical composition comprising the same, or a diagnostic agent for the individual, cell, tissue, organ, or biological fluid. In the context of a cell, administration includes contact (e.g., in vitro or ex vivo) of a reagent to the cell, as well as contact of a reagent to a fluid, where the fluid is in contact with the cell.

[0062] The terms "treat", "treating", "treatment" and the like, refer to a course of action (such as administering an IDO inhibitor or a pharmaceutical composition comprising the same) initiated following a disease, disorder or condition, or a symptom thereof, has been diagnosed, observed, and the like in order to eliminate, reduce, suppress, mitigate, or improve either temporarily or permanently, at least one of the underlying causes of a disease, disorder, or condition that afflicts an individual, or at least one of the symptoms associated with a disease, disorder, condition that afflicts an individual. Thus, treatment includes inhibition of (e.g., arresting the development or further development of the disease, disorder or condition or clinical symptoms associated therewith) of an active disease.

[0063] The term "in need of treatment" as used herein refers to a judgment made by a physician or other caregiver that an individual requires or will benefit from treatment. This judgment is made based on a variety of factors that are within the domain of the doctor or caregiver's expertise.

[0064] The terms "prevent", "preventing", "prevention" and the like refer to a course of action (such as administration of an IDO inhibitor or a pharmaceutical composition comprising the same) initiated in a manner (e.g. , prior to the onset of a disease, disorder, condition or symptom thereof) in order to prevent, suppress, inhibit or reduce, temporarily or permanently, an individual's risk of developing a disease, disorder, condition or the like (as determined, e.g. , by the absence of clinical symptoms) or delay in its onset, generally in the context of an individual predisposed to having a particular disease, disorder or condition. In certain cases, the terms also refer to slowing the progression of the disease, disorder or condition or inhibiting the progression thereof to a harmful or otherwise undesirable state.

[0065] The term "in need of prevention" as used herein refers to a judgment made by a physician or other caregiver that an individual requires or will benefit from preventative care. This judgment is made based on a variety of factors that are within the expertise of the doctor or caregiver.

[0066] The phrase "therapeutically effective amount" refers to the administration of an agent to an individual, alone or as part of a pharmaceutical composition and either in a single dose or as part of a series of doses, in an amount capable of have any positive, detectable effect on any symptom, aspect, or characteristic of a disease, disorder, or condition when administered to the individual. The therapeutically effective amount can be determined by evaluating the relevant physiological effects, and it can be adjusted in connection with the dosage regimen and diagnostic analysis of the individual's condition, and the like. By way of example, measuring the serum level of an IDO inhibitor (or, for example, a metabolite thereof) at a particular time post-administration may be indicative of whether a therapeutically effective amount was used.

[0067] The phrase "in an amount sufficient to effect a change" means that there is a detectable difference between a level of an indicator measured before (e.g., a reference level) and after administration of a particular therapy. Indicators include any objective parameter (e.g., serum concentration) or subjective parameter (e.g., an individual's sense of well-being).

[0068] The term "small molecules" refers to chemical compounds having a molecular weight that is less than about 10kDa, less than about 2kDa, or less than about 1kDa. Small molecules include, but are not limited to, inorganic molecules, organic molecules containing an inorganic component, molecules comprising a radioactive atom, and synthetic molecules. Therapeutically, a small molecule may be more permeable to cells, less susceptible to degradation, and less likely to provoke an immune response than molecules.

[0069] As used herein, the terms "IDO inhibitor", "IDO blocker" and terms similar thereto refer to agents capable of inhibiting IDO activity, thereby reversing IDO-mediated immunosuppression. An IDO inhibitor can be a competitive, non-competitive, or irreversible IDO inhibitor. "A competitive IDO inhibitor" is a compound that reversibly inhibits IDO enzyme activity at the catalytic site; "a non-competitive IDO inhibitor" is a compound that reversibly inhibits IDO enzyme activity at a non-catalytic site; and "an irreversible IDO inhibitor" is a compound that irreversibly eliminates IDO enzyme activity by forming a covalent bond (or other stable means of inhibiting enzyme function) with the enzyme. Several IDO inhibitors are commercially available (e.g., 5-Br-4-Cl-indoxyl 1,3-diacetate and 1-methyl-DL-tryptophan (1 MT); both available from Sigma-Aldrich, St. Louis, MO ) and can be used as, for example, "tool" or "reference" compounds.

[0070] The term "ligand" refers to, for example, a peptide, a polypeptide, a membrane-associated or membrane-bound molecule, or a complex thereof, which can act as an agonist or antagonist of a receptor. A ligand encompasses natural and synthetic ligands, for example, cytokines, cytokine variants, analogues, muteins, and antibody-derived binding compositions, as well as small molecules. The term also encompasses an agent that is neither an agonist nor an antagonist, but that can bind to a receptor without significantly influencing its biological properties, for example, signaling or adhesion. Furthermore, the term includes a membrane-bound ligand that has been changed, for example, by chemical or recombinant methods, to a soluble version of the membrane-bound ligand. A ligand or receptor may be entirely intracellular, that is, it may reside in the cytosol, nucleus, or some other intracellular compartment. The complex of a ligand and receptor is called a "ligand-receptor complex".

[0071] The terms "inhibitors" and "antagonists", or "activators" and "agonists" refer to inhibitory or activating molecules, respectively, for example, to the activation of, for example, a ligand, receptor , cofactor, gene, cell, tissue, or organ. Inhibitors are molecules that decrease, block, prevent, delay activation, inactivate, desensitize, or down-regulate, for example, a gene, protein, ligand, receptor, or cell. Activators are molecules that enhance, activate, facilitate, enhance activation, sensitize, or down-regulate, for example, a gene, protein, ligand, receptor, or cell. An inhibitor can also be defined as a molecule that reduces, blocks, or inactivates a constitutive activity. An "agonist" is a molecule that interacts with a target to cause or promote an increase in activation of the target. An "antagonist" is a molecule that opposes the action(s) of an agonist. An antagonist prevents, reduces, inhibits, or neutralizes the activity of an agonist, and an antagonist may also prevent, inhibit, or reduce the constitutive activity of a target, for example, a target receptor, even where there is no identified agonist.

[0072] The terms "modulate", "modulation" and the like refer to the ability of a molecule (for example, an activator or an inhibitor) to increase or decrease the function or activity of IDO, directly or indirectly. A modulator may act alone, or it may use a cofactor, for example, a protein, metal ion, or small molecule. Examples of modulators include small molecule compounds and other bio-organic molecules. Numerous libraries of small molecule compounds (e.g., combinatorial libraries) are commercially available and can function as a starting point for identifying a modulator. The skilled artisan is able to develop one or more assays (e.g., biochemical or cell-based assays) in which such compound libraries can be analyzed in order to identify one or more compounds having the desired properties; hereafter, the skilled medicinal chemist is able to optimize one or more compounds, for example, by synthesizing and evaluating analogues and derivatives thereof. Synthetic and/or molecular modeling studies can also be used to identify an activator.

[0073] The "activity" of a molecule can describe or refer to the binding of the molecule to a ligand or a receptor; to catalytic activity, the ability to stimulate gene expression or cell signaling, differentiation, or maturation; antigenic activity; modulating the activities of other molecules; and similar. The term "proliferative activity" encompasses an activity that promotes, is necessary for, or is specifically associated with, for example, normal cell division, as well as cancer, tumors, dysplasia, cellular transformation, metastasis, and angiogenesis.

[0074] As used herein, "comparable", "comparable activity", "comparable activity to", "comparable effect", "comparable effect to", and the like are relative terms that can be considered quantitatively and/or qualitatively. The meaning of terms is often dependent on the context in which they are used. By way of example, two agents that activate a receptor may be considered to have a comparable effect from a qualitative perspective, but the two agents may be considered to have no comparable effect from a quantitative perspective if one agent is only able to obtain 20% of the activity of the other agent as determined in an assay accepted in the art (e.g., a dose response assay) or in an animal model accepted in the art. When comparing a result to another result (for example, a result to a reference standard), "comparable" often (although not always) means that a result deviates from a reference standard by less than 35%, by less than than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 7%, less than 5%, less than 4%, less than 3%, less than 2%, or less than 1%. In particular embodiments, a result is comparable to a reference standard if it deviates by less than 15%, by less than 10%, or by less than 5% from the reference standard. By way of example, but without limitation, activity or effect may refer to efficacy, stability, solubility, or immunogenicity.

[0075] "Substantially pure" indicates that a component makes up more than about 50% of the total content of the composition, and typically more than about 60% of the total polypeptide content. More typically, "substantially pure" refers to compositions in which at least 75%, at least 85%, at least 90% or more of the total composition is the component of interest. In some cases, the polypeptide will make up more than about 90%, or more than about 95% of the total content of the composition.

[0076] The terms "specifically bind" or "selectively ligase", when referring to a ligand/receptor, antibody/antigen, or other binding pair, indicates a binding reaction that is determinative of the presence of the protein in a population heterogeneous array of proteins and other biologicals. Thus, under designated conditions, a specified ligand binds to a particular receptor and does not bind in a significant amount to the other proteins present in the sample. The antibody, or binding composition derived from the antigen-binding site of an antibody, of the contemplated method, binds to its antigen, or a variant or mutein thereof, with an affinity that is at least twice as high, at least ten times greater, at least 20 times greater, or at least 100 times greater than the affinity with any other antibody, or binding composition derived therefrom. In a particular embodiment, the antibody will have an affinity that is greater than about 109 liters/mol, as determined, for example, by Scatchard analysis (Munsen, et al., 1980 Analyt. Biochem. 107:220-239 ).

[0077] The term "response", for example, of a cell, tissue, organ, or organisms, encompasses a change in biochemical or physiological behavior, for example, concentration, density, adhesion, or migration within a biological compartment, rate of gene expression, or state of differentiation, where the change is correlated with activation, stimulation, or treatment, or with internal mechanisms such as genetic programming. In certain contexts, the terms "activation", "stimulation", and the like refer to cellular activation as regulated by internal mechanisms as well as external or environmental factors; while the terms "inhibition", "downregulation" and the like refer to the opposite effects.

[0078] The terms "polypeptide", "peptide", and "protein", used interchangeably herein, refer to a polymeric form of amino acids of any length, which may include genetically encoded and non-genetically encoded amino acids, or biochemically modified amino acids or derivatives, and polypeptides having modified polypeptide skeletons. Terms include fusion proteins, including, but not limited to, fusion proteins with a heterologous amino acid sequence, fusion proteins with heterologous and homologous leader sequences, with or without N-terminal methionine residues; immunologically labeled proteins; and similar.

[0079] As used herein, the terms "variants" and "homologs" are used interchangeably to refer to amino acid or DNA sequences that are similar to reference amino acid or nucleic acid sequences, respectively. The term encompasses naturally occurring variants and non-naturally occurring variants. Naturally occurring variants include homologous (polypeptides and nucleic acids that differ in amino acid or nucleotide sequence, respectively, from one species to another), and allelic variants (polypeptides and nucleic acids that differ in amino acid or nucleotide, respectively, from one individual to another within a species). Thus, variants and homologs encompass naturally occurring DNA sequences and proteins thus encoded and their isoforms, as well as binding variants of a protein or gene. The terms also encompass nucleic acid sequences that vary by one or more bases from a naturally occurring DNA sequence, but are still translated into an amino acid sequence that corresponds to the naturally occurring protein due to degeneration of the genetic code. Non-naturally occurring and homologous variants include polypeptides and nucleic acids that comprise a change in amino acid or nucleotide sequence, respectively, where the change in sequence is artificially introduced (e.g., muteins); for example, change is usually in the laboratory by human intervention ("man's hand"). Therefore, non-naturally occurring variants and homologs may also refer to those that differ from naturally occurring sequences by one or more conservative substitutions and/or tags and/or conjugates.

[0080] The term "muteins" as used herein broadly refers to mutated recombinant proteins. These proteins usually carry single or multiple amino acid substitutions and are often derived from cloned genes that have undergone site-directed or random mutagenesis, or from completely synthetic genes.

[0081] The terms "DNA", "nucleic acid", "nucleic acid molecule", "polynucleotide" and the like are used interchangeably herein to refer to a polymeric form of nucleotides of any length, deoxyribonucleotides or ribonucleotides, or analogues thereof. Non-limiting examples of polynucleotides include linear and circular nucleic acids, messenger RNA (mRNA), complementary DNA (cDNA), recombinant polynucleotides, vectors, probes, primers, and the like.

Indoleamine 2,3-Dioxygenase

[0082] As previously alluded to, IDO is an immune regulatory enzyme that is normally expressed in tumor cells and active immune cells. IDO is one of several immune response checkpoints that are involved in tumor immune escape; In this way, IDO inhibitors disrupt the mechanisms by which tumors evade the body's normal immune system.

[0083] IDO down-regulates the immune response mediated through tryptophan oxidation. This results in the inhibition of T cell activation and induction of T cell apoptosis, creating an environment in which tumor-specific cytotoxic T lymphocytes become functionally inactive or are no longer capable of attacking the individual's cancer cells. Therefore, therapeutic agents that aim to suppress tryptophan degradation by inhibiting IDO are desirable. IDO inhibitors can be used to activate T cells and therefore enhance T cell activation when T cells are suppressed by pregnancy, malignancy, or a virus such as HIV. IDO inhibition may also be an important treatment strategy for patients with neurological or neuropsychiatric diseases or disorders such as depression. The compounds, compositions, and methods herein help meet the current need for IDO modulators.

[0084] IDO expression is modulated by a complex arrangement of signals, thus implying several different mechanisms of action. For example, IDO can be induced by inhibition of DNA methyl transferases or histone deacetylases. The signaling reaction series has also been implicated in IDO function. Inhibition of NF-kB activity blocks IDO expression and produces strong antitumor responses that are both T cell and IDO dependent; alternatively, NF-KB activation (which can be accomplished by several factors such as interferon-YR1/-YR2 signaling and bell-fold receptor activation) induces IDO gene expression.

[0085] Other mechanisms are involved with the modulation of IDO function. By way of example, reactive oxidative species (ROS) inhibitors can carry out IDO stabilization; IDO levels can be modulated by inhibition or activation of reaction series that are both upstream and downstream of IDO; and activation of interferon-Y can trigger an autocrine induction of IDO.

[0086] Studies indicate that the IDO reaction series is active in most cancers, both in tumor cells as a direct defense against T cell attack, and also in antigen presenting cells (APCs) in draining lymph nodes tumor resulting in peripheral tolerance to tumor-associated antigens (TAAs). Cancers can use the IDO reaction series to facilitate the survival, growth, invasion, and metastasis of malignant cells expressing TAAs that might otherwise be recognized and attacked by the immune system.

[0087] As alluded to here, the catabolism of tryptophan in tumor tissue by the rate-limiting enzyme, IDO, provides an opportunity for the use of IDO inhibitors as a therapeutic alternative to, or an additive with, conventional chemotherapy. However, certain cancers are capable of catabolizing tryptophan but are largely IDO negative. Recent studies indicate that the reactive enzymatic reaction series of tryptophan catabolism involving tryptophan-2,3-dioxygenase (TDO) is also relevant in cancer. TDO, which is thought to be responsible for regulating systemic tryptophan levels in the liver, is constitutively expressed in some cancers and is also capable of suppressing antitumor immune responses (See, for example, Platten, M. et al., Cancer Res., 72(21):5435-5440 (1 November 2012)).

[0088] IDO is expressed in a wide variety of human tumors and tumor cell lines, as well as host APCs, which correlates with a worse clinical prognosis. Therefore, IDO inhibition may improve survival in cancer patients with IDO-mediated immunosuppression. In comparison, TDO is expressed in a wide variety of human tumors and tumor cell lines, and TDO expression is evident in advanced human glioblastomas. Identification of tumors expressing high levels of IDO or TDO may allow more selective inhibition of tryptophan-regulated immunosuppressive reaction courses. Alternatively, compounds inhibiting both IDO and TDO could provide the greatest coverage to prevent tumor escape by compensatory expression of the other tryptophan-degrading enzyme. Therefore, the use of dual IDO/TDO inhibitors or combinations of specific IDO and TDO inhibitors may prove to be a superior treatment alternative in cancer immunotherapy to block immunosuppression mediated by tryptophan metabolism.

[0089] Although a precise understanding of the underlying mechanism of action by which the compounds of the present invention carry out their activity is not required to practice the invention, the compounds (or a subgroup thereof) are believed to inhibit IDO function. Alternatively, the compounds (or a subgroup thereof) may inhibit TDO function. The compounds (or a subgroup thereof) may also have inhibitory activity on both IDO and TDO function. Although the compounds of the invention are generally referred to herein as IDO inhibitors, it is to be understood that the term "IDO inhibitors" encompasses compounds that act individually by inhibiting TDO or IDO, and/or compounds that act by inhibiting both IDO and TDO.

Identification of IDO Inhibitors Possessing Desirable Characteristics

[0090] The present invention is described, in part, for the identification of IDO inhibitors with at least one property or characteristic that is of therapeutic relevance. Candidate inhibitors can be identified using, for example, a model or assay accepted in the art, examples of which are described herein.

[0091] After identification, candidate inhibitors can also be evaluated using techniques that provide data regarding the characteristics of the inhibitors (e.g., pharmacokinetic parameters, means of determining solubility or stability). Comparisons of candidate inhibitors to a reference standard (which may be the “best in class” of current inhibitors) are indicative of the potential viability of such candidates.

Compounds of the Invention

[0092] As noted above, the present invention provides compounds represented by formula (I): or a pharmaceutically acceptable salt, hydrate or solvate thereof, wherein, the subscript n is 1 or 0; the subscript p is 1 or 0; the ring designated as A is phenyl, 5- or 6-membered heteroaryl, or C5-7 cycloalkyl; Z is O; B is N, C(OR5a), or C(R3a); each X is independently NR5a, O, CHR5, C(O), or CH(OR5a); Q is N, C(CN) or CR6; D is is a bond, O, C(R5)2, or NR5a; E is an optionally substituted 9- or 8-membered fused bicyclic heteroaryl; R1 and R2 are independently hydrogen, halogen, C1-C4 haloalkyl, C3-C6 cycloalkyl, 3- to 6-membered cycloheteroalkyl, optionally substituted phenyl, optionally substituted heteroaryl, C1-C4- optionally substituted alkyl, C1-C4 alkoxy, CN , SO2NH2, NHSO2CH3, NHSO2CF3, OCF3, SO2CH3, SO2CF3, or CONH2, and when R1 and R2 are at adjacent vertices of a phenyl ring they can be linked together to form a 5- or 6-membered cycloheteroalkyl ring having one or two ring vertices independently selected from O, N and S, wherein said cycloheteroalkyl ring is optionally substituted with one to three members selected from fluorine and C1-C3 alkyl; R3, R3a and R4 are independently hydrogen, C1-C6 optionally substituted alkyl, C2-C6 optionally substituted alkenyl, C2-C6 optionally substituted alkynyl, C1-C6 optionally substituted haloalkyl, C3-C6 cycloalkyl-C1-C4 optionally substituted alkyl, aryl -C1-C6 optionally substituted alkyl, fluorine, OH, CN, CO2H, C(O)NH2, N(R5)2, -O-C1-C6 optionally substituted alkyl, -(CR5R5)m-OH, - (CR5R5) m-CO2H, -(CR5R5)mC(O)NH2, -(CR5R5)mC(O)NHR5, - (CR5R5)mN(R5)2, -NH(CR5R5)mCO2H or -NH(CR5R5)mC(O) NH2; each R5 is independently H, F, OH, or optionally substituted C1-C6 alkyl; each R5a is independently H, or optionally substituted C1-C6 alkyl; R6 is H, OH, F, optionally substituted C1-C6 alkyl, -O-C1-C6 optionally substituted alkyl, or -N(R5a)2; and each m is independently 1, 2, or 3.

[0093] In some embodiments, Q is C(CN) or CR6.

[0094] In some embodiments, compounds are provided having the formula (Ia) or (Ib):

[0095] wherein each of the subscripts, letters, and R1, R2, R3, R4 and R6 have the meanings given with reference to the compounds of formula (I).

[0096] In other embodiments, compounds are provided having the formula (Ic):

[0097] wherein J1 is CH, N or optionally C(R2) when R2 is connected to the ring vertex identified as J1, and each of the subscripts, letters, and R1, R2, R3, R4 and R6 have the given meanings with reference to compounds of formula (I).

[0098] In some selected embodiments, compounds are provided having a formula selected from (Ic1), (Ic2), (Ic3), (Ic4), (Ic5) and (Ic6):

[0099] where J1 is CH, N or optionally C(R2) when R2 is connected to the ring vertex identified as J1, and each of the subscripts, letters, and R1, R2, R3, R3a, R4 and R6 have the meanings given with reference to compounds of formula (I).

[00100] In other embodiments, compounds are provided having the formula (Id):

[00101] where J1 is CH, N or optionally C(R2) when R2 is connected to the ring vertex identified as J1, each of the subscripts, letters, and R1, R2, R3, R3a and R6 have the meanings given with reference to compounds of formula (I).

[00102] In some selected embodiments, compounds are provided having a formula selected from (Id1), (Id2), (Id3) and (Id4):

[00103] wherein for each of (Id1) and (Id2), the compounds are provided substantially free of other isomers at the stereocenters shown. For each of (Id3) and (Id4), R11 and R12 are independently hydrogen, halogen, C1-C4 haloalkyl, C3-C6 cycloalkyl, 3- to 6-membered cycloheteroalkyl, optionally substituted phenyl, optionally substituted heteroaryl, C1-C4- optionally substituted alkyl, C1-C4 alkoxy, CN, SO2NH2, NHSO2CH3, NHSO2CF3, OCF3, SO2CH3, SO2CF3, or CONH2, and when R1 and R2 are at adjacent vertices of a phenyl ring they can be linked together to form a 5- or 6-membered cycloheteroalkyl ring having one or two ring vertices independently selected from O, N and S, wherein said cycloheteroalkyl ring is optionally substituted with one to three members selected from fluorine and C1-C3 alkyl. For each of (Id1), (Id2), (Id3) and (Id4), the remaining letters, and J1, R1, R2, R3, R3a and R6 have the meanings given with reference to the compounds of formula (Id).

[00104] In other embodiments, compounds are provided having the formula (Ie):

[00105] wherein the letters and R1, R2, R3, R3a and R6 have the meanings given with reference to the compounds of formula (I).

[00106] In some selected embodiments, compounds are provided having a formula selected from (Ie1), (Ie2), (Ie3), (Ie4), (Ie5), (Ie6) and (Ie7):

[00107] For each of the formulas (Ie1), (Ie4), (Ie5), (Ie6) and (Ie7), the compounds are substantially free of other isomers at each of the stereocenters shown. In selected embodiments of formula (Ie4), compounds are provided in which R1 is Cl, F, optionally substituted phenyl, or CN. In selected embodiments of formula (Ie6), compounds are provided in which R1 is Cl, F, optionally substituted phenyl, or CN; and R2 is H or F. In selected embodiments of formula (Ie6), compounds are provided in which R1 is Cl. In other selected embodiments of formula (Ie6), compounds are provided in which R1 is Cl; and R3 is CH3. When not specifically defined as noted herein, the remaining letters, and R1, R2, R3, R3a and R6 have the meanings given with reference to the compounds of formula (I).

[00108] In other embodiments, compounds are provided having the formula (If) or (Ig):

[00109] Each of which is substantially free of other isomers at each of the three stereocenters shown, and wherein R2 and R3 have the meanings given with reference to formula (I).

[00110] In a group of selected embodiments, any one composed of figures 1A-1L is provided.

[00111] In another group of selected embodiments, any compound of Figures 1A-1L is provided having an activity level identified as "A" or "B".

[00112] In another group of selected embodiments, any one composed of Figures 1A-1L is provided having an activity level identified as "A".

Synthesis Methods

[00113] The compounds described here can be prepared by a variety of methods. Representative methods are provided in the Examples below.

Modifications to enhance Inhibitor Traits

[00114] It is often beneficial, and sometimes imperative, to improve one or more physical properties of the treatment modalities described here and/or the manner in which they are administered. Improvements in physical properties include, for example, methods of increasing water solubility, bioavailability, serum half-life, and/or therapeutic half-life; and/or modulation of biological activity.

[00115] Known modifications in the art include pegylation, Fc fusion and albumin fusion. Although generally associated with large molecule agents (e.g., polypeptides), such modifications have recently been evaluated with particular small molecules. By way of example, Chiang, M. et al. (J. Am. Chem. Soc., 136(9):3370-3373 (2014)) describe a small molecule agonist of the adenosine 2a receptor conjugated to the immunoglobulin Fc domain. The small molecule-Fc conjugate maintained potent Fc and adenosine 2a receptor interactions and showed superior properties compared to the unconjugated small molecule. Covalent attachment of PEG molecules to small molecule therapeutics has also been described (Li, W. et al., Progress in Polymer Science, 38:421-444 (2013)).

Therapeutic and Prophylactic Uses

[00116] The present invention contemplates the use of the IDO inhibitors described herein in the treatment or prevention of a wide range of diseases, disorders and/or conditions, and/or the symptoms thereof. While particular uses are described in detail hereinafter, it should be understood that the present invention is not so limited. Furthermore, although general categories of diseases, disorders and particular conditions are mentioned hereinafter, some of the diseases, disorders and conditions may be a member of more than one category, and others may not be a member of any one category. of the categories described.

[00117] Oncology-related disorders. In accordance with the present invention, an IDO inhibitor can be used to treat or prevent a proliferative condition or disorder, including a cancer, e.g., cancer of the uterus, cervix, breast, prostate, testis, gastrointestinal tract (e.g., esophagus). , oropharynx, stomach, small or large intestines, colon, rectum), kidney, kidney cell, bladder, bone, bone marrow, skin, head or neck, liver, gallbladder, heart, lung, pancreas, salivary gland, adrenal gland, thyroid, brain (e.g., gliomas), ganglia, central nervous system (CNS), and peripheral nervous system (PNS), and cancers of the hematopoietic system and the immune system (e.g., spleen or thymus). The present invention also provides methods of treating or preventing other cancer-related diseases, disorders or conditions, including, for example, immunogenic tumors, non-immunogenic tumors, latent tumors, virus-induced cancers (e.g., epithelial cell cancers, endothelial cell carcinomas, squamous cell carcinomas and papillomavirus), adenocarcinomas, lymphomas, carcinomas, melanomas, leukemias, myelomas, sarcomas, teratocarcinomas, chemically induced cancers, metastasis, and angiogenesis. The invention contemplates reducing tolerance to a tumor cell or cancer cell antigen, for example, by modulating a regulatory T cell and/or a CD8+ T cell (see, for example, Ramirez-Montagut et al., Oncogene, 22 :3180-3187 (2003); and Sawaya et al., New Engl. J. Med., 349:1501-1509 (2003)). In particular embodiments, the tumor or cancer is colon cancer, ovarian cancer, breast cancer, melanoma, lung cancer, glioblastoma, or leukemia. The use of the term(s) "cancer-related diseases, disorders and conditions" is intended to refer broadly to conditions that are associated, directly or indirectly, with cancer, and include, for example, angiogenesis and precancerous conditions such as such as dysplasia.

[00118] In some embodiments, the present invention provides methods for treating a proliferative condition, cancer, tumor, or precancerous condition with an IDO inhibitor and at least one additional therapeutic or diagnostic agent, examples of which are mentioned in another part here.

[00119] Immune-related and inflammatory disorders. As used herein, terms such as "immune disease", "immune condition", "immune disorder", "inflammatory disease", "inflammatory condition", "inflammatory disorder" and the like are intended to broadly encompass any immune-related or inflammatory condition ( e.g., pathological inflammation and autoimmune diseases). Such conditions are often inextricably intertwined with other diseases, disorders and conditions. By way of example, an "immune condition" may refer to proliferative conditions, such as cancer, tumors, and angiogenesis; including infections (acute and chronic), tumors, and cancers that resist eradication by the immune system.

[00120] A non-limiting list of immune-related and inflammatory diseases, disorders and conditions that can be treated or prevented with the compounds and compositions of the present invention include, arthritis (e.g., rheumatoid arthritis), renal failure, lupus, asthma, psoriasis, colitis, pancreatitis, allergies, fibrosis, surgical complications (for example, where inflammatory cytokines prevent healing), anemia, and fibromyalgia. Other diseases and disorders that may be associated with chronic inflammation include Alzheimer's disease, congestive heart failure, stroke, aortic valve stenosis, arteriosclerosis, osteoporosis, Parkinson's disease, infections, inflammatory bowel disease (e.g. , Crohn's disease and ulcerative colitis), allergic contact dermatitis and other eczemas, systemic sclerosis, transplantation and multiple sclerosis.

[00121] Among other immune-related disorders, it is contemplated that inhibition of IDO function may also play a role in immunological tolerance and prevention of fetal rejection in utero.

[00122] In some embodiments, an IDO inhibitor described here can be combined with an immunosuppressive agent to reduce the number of immune effector cells.

[00123] Some of the previously mentioned diseases, disorders and conditions for which an IDO inhibitor may be particularly effective (due to, for example, limitations of current therapies) are described in greater detail here below.

[00124] Rheumatoid arthritis (RA), which is generally characterized by chronic inflammation in the membrane lining (the synovium) of the joints, affects approximately 1% of the United States population (~2.1 million people). Another understanding of the role of cytokines, including TNF-α and IL-1, in the inflammatory process has allowed the development and introduction of a new class of disease-modifying antirheumatic drugs (DMARDs). Agents (some of which overlap with treatment modalities for RA) include ENBREL® (etanercept), REMICADE® (infliximab), HUMIRA® (adalimumab), and KINERET® (anakinra). Although some of these agents relieve symptoms, inhibit the progression of structural damage, and improve physical function in particular patient populations, there is a need for alternative agents with improved efficacy, complementary mechanisms of action, and fewer/less severe adverse effects. .

[00125] Psoriasis, a constellation of common immune-mediated chronic skin diseases, affects more than 4.5 million people in the United States, of which 1.5 million are considered to have a moderate to severe form of the disease. Additionally, more than 10% of psoriasis patients develop psoriatic arthritis, which damages bone and connective tissue around joints. An improved understanding of the underlying physiology of psoriasis has resulted in the introduction of agents that, for example, target the activity of T lymphocytes and cytokines responsible for the inflammatory nature of the disease. Such agents include TNF-α inhibitors (also used in the treatment of rheumatoid arthritis (RA)), including ENBREL® (etanercept), REMICADE® (infliximab), and HUMIRA® (adalimumab)), and T cell inhibitors such as such as AMEVIVE® (alefacept) and RAPTIVA® (efalizumab). Although several of these agents are effective to some extent in certain patient populations, none have been shown to effectively treat all patients.

[00126] Individuals suffering from multiple sclerosis (MS), a seriously debilitating autoimmune disease comprising multiple areas of inflammation and myelin scarring in the brain and spinal cord, may be particularly helped by the IDO inhibitors described here, as current treatments only alleviate the symptoms or delay the progression of disability.

[00127] Similarly, IDO inhibitors may be particularly advantageous for individuals afflicted with neurodegenerative disorders, such as Alzheimer's disease (AD), a brain disorder that seriously impairs the patient's thinking, memory, and language processes; and Parkinson's disease (PD), a progressive disorder of the CNS characterized, for example, by abnormal movement, rigidity, and tremor. These disorders are progressive and debilitating, and no curative agent is available.

[00128] Virus-related disorders. The present invention contemplates the use of IDO inhibitors in the treatment and/or prevention of any disease, disorder or viral condition for which treatment with an IDO inhibitor may be beneficial. In particular embodiments, the viral disorder is a chronic viral disorder. Examples of viral diseases, disorders and conditions that are contemplated include, but are not limited to, hepatitis B virus (HBV), hepatitis C virus (HCV), human papilloma virus (HPV), HIV, AIDS (including its manifestations such as such as cachexia, dementia, and diarrhea), herpes simplex virus (HSV), Epstein-Barr virus (EBV), varicella zoster virus, coxsackie virus, and cytomegalovirus (CMV).

[00129] Disorders related to bacteria and parasites. Embodiments of the present invention contemplate administering the IDO inhibitors described herein to an individual for the treatment of a bacterial infection, e.g., a Mycobacterial infection (e.g., Mycobacterium leprae or Mycobacterium tuberculosis) or an infection caused by Listeria monocytogenes or Toxplasma gondii. Other modalities contemplate the treatment of a parasitic infection including, but not limited to, Leishmania donovani, Leishmania tropica, Leishmania major, Leishmania aethiopica, Leishmania Mexicana, Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale, or Plasmodium malariae. Often, antiparasitic therapy is administered prophylactically (for example, before an individual travels to an area with a high frequency of parasitic infection).

Pharmaceutical compositions

[00130] The IDO inhibitors of the present invention can be in the form of compositions suitable for administration to an individual. In general, such compositions are "pharmaceutical compositions" comprising an IDO inhibitor(s) and one or more pharmaceutically acceptable or physiologically acceptable diluents, carriers or excipients. In certain embodiments, IDO inhibitors are present in a therapeutically acceptable amount. Pharmaceutical compositions can be used in the methods of the present invention; Thus, for example, the pharmaceutical compositions can be administered ex vivo or in vivo to an individual in order to practice the therapeutic and prophylactic methods and uses described herein.

[00131] The pharmaceutical compositions of the present invention can be formulated to be compatible with the intended method or routine of administration; Exemplary administration routines are mentioned here. Furthermore, the pharmaceutical compositions can be used in combination with other therapeutically active agents or compounds as described herein in order to treat or prevent the diseases, disorders and conditions as contemplated by the present invention.

[00132] Pharmaceutical compositions containing the active ingredient (e.g., an inhibitor of IDO function) may be in a form suitable for oral use, for example, as tablets, capsules, troches, lozenges, aqueous or oily suspensions, dispensing powders. - persables or granules, emulsions, hard or soft capsules, or syrups, solutions, microbeads or elixirs. Pharmaceutical compositions intended for oral use may be prepared according to any method known in the art for manufacturing pharmaceutical compositions, and such compositions may contain one or more agents such as, for example, sweetening agents, flavoring agents, coloring agents and coloring agents. conservation in order to provide pharmaceutically elegant and palatable preparations. Tablets, capsules and the like contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients that are suitable for tablet manufacturing. These excipients may be, for example, diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, for example, corn starch, or alginic acid; binding agents, for example, starch, gelatin or acacia, and lubricating agents, for example, magnesium stearate, stearic acid or talc.

[00133] Tablets, capsules and the like suitable for oral administration may be uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract and thereby provide prolonged action. For example, a time delay material such as glyceryl monostearate or glyceryl distearate can be employed. They may also be coated by techniques known in the art to form osmotic therapeutic tablets for controlled release. Additional agents include biodegradable or biocompatible particles or a polymeric substance such as polyesters, polyamine acids, hydrogel, polyvinyl pyrrolidone, polyanhydrides, polyglycolic acid, ethylene vinyl acetate, methylcellulose, carboxymethylcellulose, protamine sulfate, or lactide/glycolide copolymers, copolymers of polylactide/glycolide, or ethylene vinyl acetate copolymers in order to control the release of an administered composition. For example, the oral agent may be captured in microcapsules prepared by coacervation techniques or by interfacial polymerization, by the use of hydroxymethylcellulose or gelatin microcapsules or poly(methylmethcrolate) microcapsules, respectively, or in a colloidal drug delivery system. . Colloidal dispersion systems include macromolecule complexes, nanocapsules, microspheres, microbeads, and lipid-based systems, including oil-in-water emulsions, micelles, mixed micelles, and liposomes. Methods for preparing the above-mentioned formulations will be apparent to those skilled in the art.

[00134] Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate, kaolin or microcrystalline cellulose, or as capsules of soft gelatin in which the active ingredient is mixed with water or an oily medium, for example, peanut oil, liquid paraffin, or olive oil.

[00135] Aqueous suspensions contain the active materials in mixture with suitable excipients for their manufacture. Such excipients may be suspending agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth and gum acacia; dispersing or wetting agents, e.g., a naturally occurring phosphatide (e.g., lecithin), or condensation products of an alkylene oxide with fatty acids (e.g., polyoxyethylene stearate), or condensation products of ethylene oxide with alcohols long-chain aliphatics (e.g., by heptadecaethylene-oxycetanol), or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol (e.g., polyoxyethylene sorbitol monooleate), or condensation products of ethylene- oxide with partial esters derived from fatty acids and hexitol anhydrides (e.g. polyethylene sorbitan monooleate). Aqueous suspensions may also contain one or more preservatives.

[00136] Oily suspensions can be formulated by suspending the active ingredient in a vegetable oil, for example, peanut oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. Oily suspensions may contain a thickening agent, for example beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those mentioned above, and flavoring agents can be added to provide a palatable oral preparation.

[00137] Dispersible powders and granules for preparing an aqueous suspension by adding water provide the active ingredient in mixture with a dispersing agent or wetting agent, suspending agent and one or more preservatives. Suitable dispersing agents or wetting agents are exemplified here.

[00138] The pharmaceutical compositions of the present invention can also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, for example olive oil or peanut oil, or a mineral oil, for example liquid paraffin, or mixtures thereof. Suitable emulsifying agents may be naturally occurring gums, for example acacia gum or Tragacanth gum; naturally occurring phosphatides, for example, soybeans, lecithin, and esters or partial esters derived from fatty acids; hexitol anhydrides, for example sorbitan monooleate; and condensation products of partial esters with ethylene oxide, for example, polyoxyethylene sorbitan monooleate.

[00139] Formulations may also include vehicles to protect the composition against rapid degradation or elimination from the body, such as a controlled release formulation, including implants, liposomes, hydrogels, prodrugs and microencapsulated release systems. For example, a time delay material such as glyceryl monostearate or glyceryl stearate alone, or in combination with a wax, may be employed.

[00140] Pharmaceutical compositions typically comprise a therapeutically effective amount of an IDO inhibitor contemplated by the present invention and one or more pharmaceutically and physiologically acceptable formulation agents. Suitable pharmaceutically acceptable or physiologically acceptable diluents, carriers or excipients include, but are not limited to, antioxidants (e.g., ascorbic acid and sodium bisulfate), preservatives (e.g., benzyl alcohol, methyl parabens, ethyl or n-propyl , p-hydroxybenzoate), emulsifying agents, suspending agents, dispersing agents, solvents, fillers, bulking agents, detergents, buffers, vehicles, diluents, and/or adjuvants. For example, a suitable vehicle may be physiological saline or citrate-buffered saline, possibly supplemented with other materials common in pharmaceutical compositions for parenteral administration. Neutral buffered saline or mixed saline with serum albumin are also exemplary vehicles. Those skilled in the art will readily recognize a variety of buffers that can be used in the pharmaceutical compositions and dosage forms contemplated herein. Typical buffers include, but are not limited to, pharmaceutically acceptable weak acids, weak bases, or mixtures thereof. As an example, the buffer components may be water-soluble materials such as phosphoric acid, tartaric acid, lactic acid, succinic acid, citric acid, acetic acid, ascorbic acid, aspartic acid, glutamic acid, and salts thereof. Acceptable buffering agents include, for example, a Tris buffer, N-(2-Hydroxyethyl)piperazine-N'-(2-ethanesulfonic acid) (HEPES), 2-(N-morpholino)ethanesulfonic acid (MES), salt of sodium 2-(N-morpholino)ethanesulfonic acid (MES), 3-(N-morpholino)propanesulfonic acid (MOPS), and N-tris[hydroxymethyl]methyl-3-aminopropanesulfonic acid (TAPS).

[00141] After a pharmaceutical composition has been formulated, it can be stored in sterile vials as a solution, suspension, gel, emulsion, solid, or dehydrated or lyophilized powder. Such formulations may be stored in a ready-to-use form, a lyophilized form requiring reconstitution before use, a liquid form requiring dilution before use, or another acceptable form. In some embodiments, the pharmaceutical composition is provided in a single-use container (e.g., a single-use vial, ampoule, syringe, or auto-injector (similar to, e.g., an EPIPEN®)), while a multi-use container (e.g., a multi-use bottle) is provided in other embodiments. Any drug delivery apparatus can be used to release the IDO inhibitor, including implants (e.g., implantable pumps) and catheter systems, pumps and slow injection devices, all of which are well known to the skilled artisan. Depot injections, which are generally administered subcutaneously or intramuscularly, can also be used to release the polypeptides described herein over a defined period of time. Depot injections are generally solid or oil based and generally comprise at least one of the formulation components mentioned herein. One skilled in the art is familiar with formulations and possible uses of depot injections.

[00142] Pharmaceutical compositions can be in the form of a sterile injectable aqueous or oily suspension. This suspension may be formulated in accordance with the known art using those suitable dispersing agents or wetting agents or suspending agents mentioned herein. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parenterally acceptable diluent or solvent, for example, as a solution in 1,3-butane diol. Acceptable diluents, solvents and dispersion media that may be employed include water, Ringer's solution, isotonic sodium chloride solution, CREMOPHOR® EL (BASF, Parsippany, NJ) or phosphate buffered saline (PBS), ethanol, polyol (e.g., glycerol, propylene glycol, and liquid polyethylene glycol), and suitable mixtures thereof. Furthermore, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose, any mild fixed oil may be employed, including synthetic mono- or diglycerides. Furthermore, fatty acids such as oleic acid find use in the preparation of injectables. Prolonged absorption of particular injectable formulations can be achieved by including an agent that delays absorption (e.g., aluminum monostearate or gelatin).

[00143] The present invention contemplates the administration of IDO inhibitors in the form of suppositories for rectal administration. Suppositories can be prepared by mixing the drug with a suitable non-irritating excipient that is solid at ordinary temperatures but liquid at rectal temperature and will therefore melt in the rectum to release the drug. Such materials include, but are not limited to, cocoa butter and polyethylene glycols.

[00144] The IDO inhibitors contemplated by the present invention may be in the form of any other pharmaceutical composition (for example, sprays for nasal use or inhalation) currently known or developed in the future.

[00145] The concentration of a polypeptide or fragment thereof in a formulation can vary widely (e.g., from less than about 0.1%, generally at or at least about 2% to as much as 20% to 50% or more by weight) and will generally be selected primarily based on fluid volumes, viscosities, and individual-based factors according to, for example, the particular mode of administration selected.

Administration Routines

[00146] The present invention contemplates the administration of IDO inhibitors, and compositions thereof, in any suitable manner. Suitable administration routines include oral, parenteral (e.g., intramuscular, intravenous, subcutaneous (e.g., injection or implant), intraperitoneal, intracisternal, intraarticular, intraperitoneal, intracerebral (intraparenchymal), and intracerebroventricular), nasal, vaginal, sublingual, intraocular, rectal, topical (e.g., transdermal), sublingual, and inhalation. Depot injections, which are generally administered subcutaneously or intramuscularly, can also be used to release the IDO inhibitors described here over a defined period of time.

[00147] Particular modalities of the present invention include oral administration.

Combination Therapy

[00148] The present invention contemplates the use of IDO inhibitors in combination with one or more active therapeutic agents (e.g., chemotherapeutic agents) or other therapeutic modalities (e.g., radiation). In such combination therapy, the various active agents often have different complementary mechanisms of action. Such combination therapy may be especially advantageous by allowing a dose reduction of one or more of the agents, thereby reducing or eliminating adverse effects associated with one or more of the agents. Furthermore, such combination therapy may have a synergistic therapeutic or prophylactic effect on the underlying disease, disorder or condition.

[00149] As used herein, "combination" is intended to include therapies that can be administered separately, e.g., separately formulated for separate administration (e.g., as may be provided in a kit), and therapies that can be administered together in a simple formulation (i.e., a "co-formulation").

[00150] In certain embodiments, IDO inhibitors are administered or applied sequentially, for example, where one agent is administered before one or more other agents. In other embodiments, IDO inhibitors are administered simultaneously, for example, where two or more agents are administered at or about the same time; The two or more agents may be present in two or more separate formulations or combined in a single formulation (i.e., a coformulation). Regardless of whether the two or more agents are administered sequentially or simultaneously, they are considered to be administered in combination for the purposes of the present invention.

[00151] The IDO inhibitors of the present invention can be used in combination with at least one other (active) agent in any manner appropriate under the circumstances. In one embodiment, treatment with at least one active agent and at least one IDO inhibitor of the present invention is maintained for a period of time. In another embodiment, treatment with at least one active agent is reduced or discontinued (e.g., when the subject is stable), while treatment with an IDO inhibitor of the present invention is maintained at a constant dosage regimen. In another embodiment, treatment with at least one active agent is reduced or discontinued (e.g., when the subject is stable), while treatment with an IDO inhibitor of the present invention is reduced (e.g., lower dose, less dosage). frequent or shorter treatment regimen). In yet another embodiment, treatment with at least one active agent is reduced or discontinued (e.g., when the subject is stable), and treatment with the IDO inhibitor of the present invention is increased (e.g., higher dose, higher dosage). frequent or longer treatment regimen). In yet another embodiment, treatment with at least one active agent is continued and treatment with the IDO inhibitor of the present invention is reduced or discontinued (e.g., lower dose, less frequent dosing, or shorter treatment regimen). In yet another embodiment, treatment with at least one active agent and treatment with the IDO inhibitor of the present invention are reduced or discontinued (e.g., lower dose, less frequent dosing, or shorter treatment regimen).

[00152] Oncology-related disorders. The present invention provides methods for treating and/or preventing a proliferative condition, cancer, tumor, or precancerous disease, disorder, or condition with an IDO inhibitor and at least one additional therapeutic agent, such as radiation, an immunomodulatory or chemotherapeutic agent, or diagnostic agent. Suitable immunomodulatory agents that can be used in the present invention include CD40L, B7, and B7RP1; activating monoclonal antibodies (mAbs) to stimulatory receptors, such as anti-CD40, anti-CD38, anti-ICOS, and 4-IBB ligand; dendritic cell antigen loading (in vitro or in vivo); anticancer vaccines such as dendritic cell cancer vaccine; cytokines/chemokines, such as IL1, IL2, IL12, IL18, ELC/CCL19, SLC/CCL21, MCP-1, IL-4, IL-18, TNF, IL-15, MDC, IFNa/b, M-CSF , IL-3, GM-CSF, IL-13, and anti-IL-10; bacterial lipopolysaccharides (LPS); and immunostimulatory oligonucleotides.

[00153] Examples of chemotherapeutic agents include, but are not limited to, alkylating agents such as thiotepa and cyclophosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethyleneimines and methylamelamines including altretamine, triethylenemelamine, triethylenephosphoramide, triethylenethiophosphaoramide and trimethylomelamime; nitrogen mustards such as chiorambucil, chlornafazine, colophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembicine, phenesterine, prednimustine, trophosphamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, ranimustine; antibiotics such as aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, calicheamicin, carabicin, caminomycin, carzinophylline, chromomycins, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin, epirubicin, esorubicin , idarubicin, marcellomycin, mitomycins, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, chelamycin, rhodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; antimetabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogues such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogues such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine, 5-FU; androgens such as calusterone, dromostanolone propionate, epithiostanol, mepitiostane, testolactone; antiadrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; ansacrine; bestrabucil; bisantrene; edatraxate; desphophamine; demecolcine; diaziquone; elformitin; elliptinium acetate; etoglucide; gallium nitrate; hydroxyurea; lentinan; lonidamine; mitoguazone; mitoxantrone; mopidamol; nitracrine; pentostatin; fenamet; pirarubicin; podophyllinic acid; 2-ethylhydrazide; procarbazine; razoxane; sizofiran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2''- trichlorotriethylamine; urethane; vindesina; dacarbazine; manomustine; mito-bronitol; mitolactol; pipobroman; gacytosin; arabinoside (Ara-C); cyclophosphamide; thiotepa; taxoids, e.g. paclitaxel and doxetaxel; chlorambucil; gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum and platinum coordination complexes such as cisplatin and carboplatin; vinblastine; etoposide (VP-16); ifosfamide; mitomycin C; mitoxantrone; vincristine; vinorelbine; navelbina; novantrona; teniposide; daunomycin; aminopterin; xeloda; ibandronate; CPT11; topoisomerase inhibitors; difluoromethylornithine (DMFO); retinoic acid; esperamycins; capecitabine; and pharmaceutically acceptable salts, acids and derivatives of any of the above.

[00154] Chemotherapeutic agents also include antihormonal agents that act to regulate or inhibit hormonal action in tumors such as antiestrogens, including, for example, tamoxifen, raloxifene, aromatase inhibiting 4(5)-imidazoles, 4-hydroxytamoxifen, trioxyfen, ceoxifen, onapristone, and toremifene; and antiandrogens such as flutamide, nilutamide, bicalutamide, leuprolide, and goserelin; and pharmaceutically acceptable salts, acids or derivatives of any of the above. In certain embodiments, combination therapy comprises administering a hormone or related hormonal agent.

[00155] Chemotherapeutic agents also include signal transduction inhibitors (STI). The term "signal transduction inhibitor" refers to an agent that selectively inhibits one or more steps in a series of signaling reactions. Signal transduction inhibitors (STIs) of the present invention include: (i) bcr/abl kinase inhibitors (e.g., GLEEVEC); (ii) epidermal growth factor (EGF) receptor inhibitors, including kinase inhibitors and antibodies; (iii) her-2/neu receptor inhibitors (e.g., HERCEPTIN); (iv) inhibitors of Akt family kinases or the Akt reaction series (e.g., rapamycin); (v) cell cycle kinase inhibitors (e.g., flavopiridol); and (vi) phosphatidyl inositol kinase inhibitors.

[00156] Additional treatment modalities that can be used in combination with an IDO inhibitor include a cytokine or cytokine antagonist, such as IL-12, IFN, or anti-epidermal growth factor receptor, radiotherapy, a monoclonal antibody against IDO antigen tumor, a complex of a monoclonal antibody and toxin, a T cell adjuvant, bone marrow transplant, or antigen-presenting cells (e.g., dendritic cell therapy). Vaccines (e.g., as a soluble protein or as nucleic acid encoding the protein) are also provided herein.

[00157] Cardiovascular Diseases. The present invention provides methods for treating and/or preventing certain cardiovascular and/or metabolically related diseases, disorders and conditions, as well as disorders associated therewith, with an IDO inhibitor and at least one therapeutic or diagnostic agent. additional.

[00158] Examples of therapeutic agents useful in combination therapy for the treatment of hypercholesterolemia (and also atherosclerosis) include statins (e.g., CRESTOR, LESCOL, LI-PITOR, MEVACOR, PRAVACOL, and ZOCOR), which inhibit the synthesis cholesterol enzyme; bile acid resins (e.g., COLESTID, LO-CHOLEST, PREVALITE, QUESTRAN, and WELCHOL), which sequester cholesterol and prevent its absorption; ezetimibe (ZETIA), which blocks cholesterol absorption; fibric acid (e.g., TRICOR), which lowers triglycerides and may modestly increase HDL; niacin (e.g., NIACOR), which modestly lowers LDL cholesterol and triglycerides; and/or a combination of the previously mentioned (e.g., VYTORIN (ezetimibe with simvastatin). Alternative cholesterol treatments that may be candidates for use in combination with the IDO inhibitors described herein include various supplements and herbs (e.g., garlic, policosanol , and gug-gul). The present invention encompasses pharmaceutically acceptable salts, acids or derivatives of any of the above.

[00159] Immune-related and Inflammatory Disorders. The present invention provides methods for treating and/or preventing immune-related and/or inflammatory diseases, disorders and conditions, as well as disorders associated therewith, with an IDO inhibitor and at least one additional therapeutic or diagnostic agent.

[00160] Examples of therapeutic agents useful in combination therapy include, but are not limited to, the following: non-steroidal anti-inflammatory drug (NSAID) such as aspirin, ibuprofen, and other propionic acid derivatives (alminoprofen, benoxapro - hay, bucloxic acid, carprofen, fenbufen, fenoprofen, fluprofen, flurbiprofen, indoprofen, ketoprofen, miroprofen, naproxen, oxaprozin, pirprofen, pranoprofen, suprofen, tiaprofenic acid, and tioxaprofen), acetic acid derivatives (indomethacin, acemethacin, alclofenac, clidanac, diclofenac, fenclofenac, fenclozic acid, fentiazac, Fuirofenac, ibufenac, isoxepac, oxpinac, sulindac, tiopinac, tolmetin, zidomethacin, and zomepirac), fenamic acid derivatives (flufenamic acid, meclofenamic acid, mefenamic acid , niflumic acid and tolfenamic acid), biphenylcarboxylic acid derivatives (diflunisal and flufenisal), oxicans (isoxicam, piroxicam, sudoxicam and tenoxicam), salicylates (acetyl salicylic acid, sulfasalazine) and pyrazolones (apazone, bezpiperilon, feprazone, mofebutazone , oxyphenbutazone, phenylbutazone). Other combinations include cyclooxygenase-2 (COX-2) inhibitors.

[00161] Other active agents for combination include steroids such as prednisolone, prednisone, methylprednisolone, betamethasone, dexamethasone, or hydrocortisone. Such a combination can be especially advantageous since one or more adverse effects of the steroid can be reduced or even eliminated by decreasing the dose of steroid required.

[00162] Additional examples of active agents that can be used in combinations to treat, for example, rheumatoid arthritis, include cytokine suppressive anti-inflammatory drug(s) (CSAIDs); antibodies to, or antagonists of, other human cytokines or growth factors, e.g., TNF, LT, IL-1β, IL-2, IL-6, IL-7, IL-8, IL-15, IL-16, IL-18, EMAP-II, GM-CSF, FGF, or PDGF.

[00163] Particular combinations of active agents can interfere at different points in the autoimmune and subsequent inflammatory cascade, and include TNF antagonists such as human or humanized, chimeric TNF antibodies, REMICADE, anti-TNF antibody fragments (e.g., CDP870) , and soluble p55 or p75 TNF receptors, derivatives thereof, p75TNFRIgG (ENBREL.) or p55TNFR1gG (LENERCEPT), soluble IL-13 receptor (sIL-13), and also TNFα-converting enzyme inhibitors (TACE); Similarly, IL-1 inhibitors (e.g., Interleukin-1 converting enzyme inhibitors) may be effective. Other combinations include Interleukin 11, anti-P7s glycoprotein ligand, and p-selectin (PSGL). Other examples of agents useful in combination with the IDO inhibitors described herein include interferon-β1a (AVONEX); interferon- β1b (BETASERON); copaxone; hyperbaric oxygen; intravenous immunoglobulin; clabribine; and antibodies to, or antagonists of, other human cytokines or growth factors (e.g., antibodies to CD40 and CD80 ligand).

[00164] Immune checkpoint inhibitors. The present invention contemplates the use of the IDO function inhibitors described herein in combination with additional immune checkpoint inhibitors.

[00165] The enormous number of genetic and epigenetic changes that are characteristic of all cancers provide a diverse array of antigens that the immune system can use to distinguish tumor cells from their normal counterparts. In the case of T cells, the ultimate breadth (e.g., levels of cytokine production or proliferation) and quality (e.g., the type of immune response generated, such as the pattern of cytokine production) of the response, which is initiated through of antigen recognition by the T cell receptor (TCR), is regulated by a balance between costimulatory and inhibitory signals (immunological checkpoints). Under normal physiological conditions, immunological checkpoints are crucial for the prevention of autoimmunity (i.e., the maintenance of self-tolerance) and also for the protection of tissues from damage when the immune system is responding to pathogenic infection. The expression of immune checkpoint proteins can be dysregulated by tumors as an important immune resistance mechanism.

[00166] T cells have been the main focus of efforts to therapeutically manipulate endogenous antitumor immunity because of i) their ability for selective recognition of protein-derived peptides in all cellular compartments; ii) its ability to directly recognize and exterminate cells expressing antigens (by CD8+ effector T cells; also known as cytotoxic T lymphocytes (CTLs)); and iii) its ability to orchestrate diverse immune responses by CD4+ helper T cells, which integrate adaptive and innate effector mechanisms. In clinical routine, immune checkpoint blockade - which results in amplification of antigen-specific T cell responses - has shown to be a promising method in human cancer therapeutics.

[00167] T cell-mediated immunity includes multiple sequential steps, each of which is regulated by counterbalance and inhibitory signals in order to optimize the response. While almost all inhibitory signals in the immune response ultimately modulate intracellular signaling reaction series, many are initiated through membrane receptors, the ligands of which are membrane-bound or soluble (cytokines). While costimulatory and inhibitory receptors and ligands that regulate T cell activation are often not overexpressed in cancers relative to normal tissues, inhibitory ligands and receptors that regulate T cell effector functions in tissues are commonly overexpressed in tumor cells or in non-transformed cells associated with the tumor microenvironment. The functions of the membrane-bound and soluble receptor - immunological checkpoint ligand can be modulated using agonist antibodies (for co-stimulatory reaction sets) or antagonist antibodies (for inhibitory reaction sets). Thus, unlike most antibodies currently approved for cancer therapy, antibodies that block immune checkpoints do not target tumor cells directly, but rather target lymphocyte receptors or their ligands in order to enhance endogenous antitumor activity. [See Pardoll, (April 2012) Nature Rev. Cancer 12:252-64].

[00168] Examples of immune checkpoints (ligands and receptors), some of which are selectively upregulated on various types of tumor cells, that are candidates for blockade include PD1 (programmed cell death protein 1); PDL1 (PD1 ligand); BTLA (B and T lymphocyte attenuator); CTLA4 (cytotoxic T lymphocyte-associated antigen 4); TIM3 (T cell membrane protein 3); LAG3 (lymphocyte activation gene 3); A2aR (adenosine receptor A2a A2aR); and Killer inhibitory receptors, which can be divided into two classes based on their structural characteristics: i) killer cell immunoglobulin-like receptors (KIRs), and ii) C-type lectin receptors (members of the type II transmembrane receptor family) . Other less well-defined immunological checkpoints have been described in the literature, including both receptors (e.g., the 2B4 receptor (also known as CD244)) and ligands (e.g., certain inhibitory ligands of the B7 family such as B7- H3 (also known as CD276) and B7- H4 (also known as B7-S1, B7x and VCTN1)). [See Pardoll, (April 2012) Nature Rev. Cancer 12:252-64].

[00169] The present invention contemplates the use of the IDO function inhibitors described herein in combination with inhibitors of the previously mentioned immune checkpoint receptors and ligands, as well as immune checkpoint receptors and ligands yet to be described. Certain immune checkpoint modulators are currently available, while others are in late-stage development. To illustrate, when it was approved for the treatment of melanoma in 2011, the fully humanized CTLA4 monoclonal antibody ipilimumab (YERVOY; Bristol-Myers Squibb) became the first immune checkpoint inhibitor to receive regulatory approval in the United States. . Fusion proteins comprising CTLA4 and an antibody (CTLA4-Ig; abatcept (ORENCIA; Bristol-Myers Squibb)) have been used for the treatment of rheumatoid arthritis, and other fusion proteins have been shown to be effective in kidney transplant patients who are sensitive to Epstein Barr Virus. PD1 antibodies are also available for the treatment of cancer, including, for example, nivolumab (Bristol-Myers Squibb) and pembroluzimab (Merck), and anti-PDL1 antibodies are also being evaluated (e.g., MPDL3280A (Roche)). . Nivolumab (Opdivo®) has shown promise in patients with melanoma, lung and kidney cancer, as well as multiple other malignancies.

[00170] In one aspect of the present invention, the claimed IDO inhibitors are combined with an immuno-oncology agent that is (i) an agonist of a stimulatory receptor (including a co-stimulatory) or (ii) an antagonist of an inhibitory signal (including a coinhibitory one) on T cells, both of which result in amplification of antigen-specific T cell responses. Certain stimulatory and inhibitory molecules are members of the super immunoglobulin (IgSF) family. An important of membrane-bound ligands that bind to co-stimulatory or co-stimulatory receptors is the B7 family, which includes B7-1, B7-2, B7-H1 (PD-L1), B7-DC (PD-L2), B7- H2 (ICOS-L), B7-H3, B7-H4, B7-H5 (VISTA), and B7-H6. Another family of membrane-bound ligands that bind to costimulatory or co-inhibitory receptors is the TNF family of molecules that bind to cognate TNF receptor family members, which include CD40 and CD40L, OX-40, OX-40L, CD70, CD27L, CD30, CD30L, 4-1BBL, CD137 (4-1BB), TRAIL/Apo2-L, TRAILR1/DR4, TRAILR2/DR5, TRAILR3, TRAILR4, OPG, RANK, RANKL, TWEAKR/Fn14, TWEAK, BAFFR, EDAR , XE- DAR, TACI, APRIL, BCMA, LTβR, LIGHT, DcR3, HVEM, VEGI/TL1A, TRAMP/DR3, EDAR, EDA1, 1β2, FAS, FASL, RELT, DR6, TROY, NGFR.

[00171] In another aspect, the immuno-oncology agent is a cytokine that inhibits T cell activation (e.g., IL-6, IL-10, TGF-β, VEGF, and other immunosuppressive cytokines) or a cytokine that stimulates T cell activation to stimulate an immune response.

[00172] In one aspect, T cell responses can be stimulated by a combination of the claimed IDO inhibitors and one or more of (i) an antagonist of a protein that inhibits T cell activation (e.g., point inhibitors). immunological check) such as CTLA-4, PD-1, PD-L1, PD-L2, LAG-3, TIM-3, Galectin 9, CEACAM-1, BTLA, CD69, Galectin-1, TIGIT, CD113 , GPR56, VISTA, 2B4, CD48, GARP, PD1H, LAIR1, TIM-1, and TIM-4, and/or (ii) an agonist of a protein that stimulates T cell activation such as B7-1, B7-2, CD28, 4-1BB (CD137), 4-1BBL, ICOS, ICOS-L, OX40, OX40L, GITR, GITRL, CD70, CD27, CD40, DR3 and CD2. Other agents that can be combined with the IDO inhibitors of the present invention for the treatment of cancer include antagonists of inhibitory receptors on NK cells or agonists of activating receptors on NK cells. For example, the compounds herein can be combined with KIR antagonists, such as lirilumab.

[00173] Still other agents for combination therapies include agents that inhibit or deplete macrophages or monocytes, including but not limited to CSF-1R antagonists such as CSF-1R antagonist antibodies including RG7155 (WO11/70024, WO11/107553, WO11/131407, WO13/87699, WO13/119716, WO13/132044) or FPA-008 (WO11/140249; WO13169264; WO14/036357).

[00174] In another aspect, the claimed IDO inhibitors can be used with one or more of agonistic agents that bind positive costimulatory receptors, blocking agents that attenuate signaling through inhibitory receptors, antagonists, and one or more agents that systemically increase the frequency of antitumor T cells, agents that overcome distinct immunosuppressive reaction sets within the tumor microenvironment (e.g., blockade of inhibitory receptor engagement (e.g., PD-L1/PD-1 interactions), deplete or inhibit Tregs (e.g., using an anti-CD25 monoclonal antibody (e.g., daclizumab) or by ex vivo anti-CD25 bead depletion), or reverse/prevent T cell anergy or exhaustion) and agents that cause activation innate immune and/or inflammation at tumor sites.

[00175] In one aspect, the immuno-oncology agent is a CTLA-4 antagonist, such as an antagonistic CTLA-4 antibody. Suitable CTLA-4 antibodies include, for example, YERVOY (ipilimumab) or tremelimumab.

[00176] In another aspect, the immuno-oncology agent is a PD-1 antagonist, such as an antagonistic PD-1 antibody. Suitable PD-1 antibodies include, for example, OPDIVO (nivolumab), KEYTRUDA (pembrolizumab), or MEDI-0680 (AMP-514; WO2012/145493). The immuno-oncology agent may also include pidilizumab (CT-011), although its specificity regarding PD-1 binding has been questioned. Another method to target the PD-1 receptor is the recombinant protein composed of the extracellular domain of PD-L2 (B7-DC) fused to the Fc portion of IgG1, called AMP-224

[00177] In another aspect, the immuno-oncology agent is a PD-L1 antagonist, such as an antagonistic PD-L1 antibody. Suitable PD-L1 antibodies include, for example, MPDL3280A (RG7446; WO2010/077634), durvalumab (MEDI4736), BMS-936559 (WO2007/005874), and MSB0010718C (WO2013/79174).

[00178] In another aspect, the immuno-oncology agent is a LAG-3 antagonist, such as an antagonistic LAG-3 antibody. Suitable LAG3 antibodies include, for example, BMS-986016 (WO10/19570, WO14/08218), or IMP-731 or IMP-321 (WO08/132601, WO09/44273).

[00179] In another aspect, the immuno-oncology agent is a CD137 (4-1BB) agonist, such as an agonistic CD137 antibody. Suitable CD137 antibodies include, for example, urelumab and PF-05082566 (WO12/32433).

[00180] In another aspect, the immuno-oncology agent is a GITR agonist, such as an agonistic GITR antibody. Suitable GITR antibodies include, for example, BMS-986153, BMS-986156, TRX-518 (WO06/105021, WO09/009116) and MK-4166 (WO11/028683).

[00181] In another aspect, the immuno-oncology agent is an OX40 agonist, such as an agonistic OX40 antibody. Suitable OX40 antibodies include, for example, MEDI-6383 or MEDI-6469.

[00182] In another aspect, the immuno-oncology agent is an OX40L antagonist, such as an antagonistic OX40 antibody. Suitable OX40L antagonists include, for example, RG-7888 (WO06/029879).

[00183] In another aspect, the immuno-oncology agent is a CD40 agonist, such as an agonistic CD40 antibody. In yet another embodiment, the immuno-oncology agent is a CD40 antagonist, such as an antagonistic CD40 antibody. Suitable CD40 antibodies include, for example, lucatumumab or dacetuzumab.

[00184] In another aspect, the immuno-oncology agent is a CD27 agonist, such as an agonistic CD27 antibody. Suitable CD27 antibodies include, for example, varlilumab.

[00185] In another aspect, the immuno-oncology agent is MGA271 (a B7H3) (WO11/109400).

[00186] The present invention encompasses pharmaceutically acceptable salts, acids or derivatives of any of the above.

[00187] Viral Diseases. The present invention provides methods for treating and/or preventing viral diseases, disorders and conditions, as well as disorders associated therewith, with an IDO inhibitor and at least one additional therapeutic or diagnostic agent (e.g., one or more other antiviral agents and/or one or more agents not associated with viral therapy).

[00188] Such combination therapy includes antiviral agents targeting various stages of the life cycle and having different mechanisms of action, including, but not limited to, the following: viral uncoating inhibitors (e.g., amantadine and rimantidine); reverse transcriptase inhibitors (e.g., acyclovir, zidovudine, and lamivudine); agents that target integrase; agents that block the binding of transcription factors to viral DNA; agents (e.g., antisense molecules) that impact translation (e.g., fomivirsen); agents that modulate ribozyme translation/function; protease inhibitors; viral assembly modulators (e.g., rifampicin); antiretrovirals such as, for example, nucleoside analogue reverse transcriptase inhibitors (e.g., azidothymidine (AZT), ddl, ddC, 3TC, d4T); non-nucleoside reverse transcriptase inhibitors (e.g., efavirenz, nevirapine); nucleotide analogue reverse transcription inhibitors; and agents that prevent the release of viral particles (for example, zanamivir and oseltamivir). Treatment and/or prevention of certain viral infections (e.g., HIV) often involves a group ("cocktail") of antiviral agents.

[00189] Other antiviral agents contemplated for use in combination with an IDO inhibitor include, but are not limited to, the following: abacavir, adefovir, amantadine, amprenavir, ampligen, arbidol, atazanavir, atripla, boceprevirertet, cidofovir, combivir, darunavir , delavirdine, didanosine, docosanol, edoxudine, emtricitabine, enfuvirtide, entecavir, famciclovir, fosamprenavir, foscarnet, phosphonet, ganciclovir, ibacitabine, immunovir, idoxuridine, imiquimod, indinavir, inosine, various interferons (e.g., peginterferon alfa-2a) , lopinavir, loviride, maraviroc, moroxidine, metisazone, nelfinavir, nexavir, penciclovir, peramivir, pleconaril, podophyllotoxin, raltegravir, ribavirin, ritonavir, pyramidine, saquinavir, stavudine, telaprevir, tenofovir, tipranavir, trifluridine, trizivir , tromantadine, truvada, valacyclovir, valganciclovir, vicriviroc, vidarabine, viraidine, and zalcitabine.

[00190] The present invention encompasses pharmaceutically acceptable salts, acids or derivatives of any of the above.

[00191] Parasitic Disorders. The present invention contemplates the use of the IDO function inhibitors described herein in combination with antiparasitic agents. Such agents include, but are not limited to, thiabendazole, pyrantel pamoate, mebendazole, praziquantel, niclosamide, bitionol, oxaniquine, metrifonate, ivermectin, albendazole, eflornithine, melarsoprol, pentamidine, benznidazole, nifurtimox, and nitroimidazole. The skilled artisan is aware of other agents that may find use in the treatment of parasitic disorders.

[00192] The present invention encompasses pharmaceutically acceptable salts, acids or derivatives of any of the above.

[00193] Bacterial Infections. Embodiments of the present invention contemplate the use of the IDO inhibitors described herein in combination with agents useful in the treatment or prevention of bacterial disorders. Antibacterial agents can be classified in several ways, including based on mechanism of action, based on chemical structure, and based on spectrum of activity. Examples of antibacterial agents include those that target the bacterial cell wall (e.g., cephalosporins and penicillins) or the cell membrane (e.g., polymyxins), or interfere with essential bacterial enzymes (e.g., sulfonamides, rifamycins, and quinolines) . Most antibacterial agents that target protein synthesis (e.g., tetracyclines and macrolides) are bacteriostatic, while agents such as aminoglycosides are bactericidal. Other methods of categorizing antibacterial agents are based on their target specificity; "Narrow-spectrum" agents target different types of bacteria (e.g., gram-bacteria such as streptococcus), while "broad-spectrum" agents have activity against a broad range of bacteria. The skilled artisan is aware of the types of antibacterial agents that are appropriate for use in specific bacterial infections.

[00194] The present invention encompasses pharmaceutically acceptable salts, acids or derivatives of the agents (and members of the classes of agents) mentioned above.

Dosage

[00195] The IDO inhibitors of the present invention can be administered to an individual in an amount that is dependent, for example, on the objective of administration (for example, the degree of resolution desired); the age, weight, sex, and health and physical condition of the individual to whom the formulation is being administered; the administration routine; and the nature of the disease, disorder, condition or symptom thereof. The dosage regimen may also take into account the existence, nature, and extent of any adverse effects associated with the agent(s) being administered. Effective dosage amounts and dosage regimens can readily be determined from, for example, dose escalation and safety experiments, in vivo studies (e.g., animal models), and other methods known to the skilled artisan.

[00196] In general, dosage parameters dictate that the dosage amount be less than an amount that may be irreversibly toxic to the individual (the maximum tolerated dose (MTD)) and not less than an amount required to produce an effect measurable in an individual. Such quantities are determined, for example, by the pharmacokinetic and pharmacodynamic parameters associated with ADME, taking into account the administration routine and other factors.

[00197] An effective dose (ED) is the dose or amount of an agent that produces a therapeutic response or desired effect in some fraction of the individuals ingesting it. The "median effective dose" or ED50 of an agent is the dose or amount of an agent that produces a therapeutic response or desired effect in 50% of the population to which it is administered. Although the ED50 is commonly used as a measure of reasonable expectation of an agent's effect, it is not necessarily the dose that a clinician would consider appropriate, taking into account all relevant factors. Thus, in some situations, the effective amount is greater than the calculated ED50, in other situations, the effective amount is less than the calculated ED50, and in still other situations, the effective amount is equal to the calculated ED50.

[00198] Furthermore, an effective dose of the IDO inhibitors of the present invention can be an amount that, when administered in one or more doses to an individual, produces a desired result with respect to a healthy individual. For example, for an individual experiencing a particular disorder, an effective dose may be one that improves a diagnostic parameter, measure, marker, and the like of that disorder by at least about 5%, at least about 10%, at least about 20%. %, at least about 25%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80% , at least about 90%, or more than 90%, where 100% is defined as the diagnostic parameter, measurement, marker and the like exhibited by a normal individual.

[00199] For administration of an oral agent, the compositions can be provided in the form of tablets, capsules and the like containing from 1.0 to 1000 milligrams of the active ingredient, particularly 1.0, 3.0, 5.0, 10, 0, 15.0, 20.0, 25.0, 50.0, 75.0, 100.0, 150.0, 200.0, 250.0, 300.0, 400.0, 500.0, 600.0, 750.0, 800.0, 900.0, and 1000.0 milligrams of the active ingredient.

[00200] In certain embodiments, the dosage of the desired IDO inhibitor is contained in a "unit dosage form". The phrase "unit dosage form" refers to physically discrete units, each unit containing a predetermined amount of the IDO inhibitor, alone or in combination with one or more additional agents, sufficient to produce the desired effect. It will be appreciated that the parameters of a unit dosage form depend on the particular agent and the effect to be obtained.

Kits

[00201] The present invention also contemplates kits comprising an IDO inhibitor, and pharmaceutical compositions thereof. Kits are generally in the form of a physical structure housing several components, as described below, and can be used, for example, in practicing the methods described above.

[00202] A kit may include one or more of the IDO inhibitors described herein (provided and, for example, a sterile container), which may be in the form of a pharmaceutical composition suitable for administration to an individual. IDO inhibitors may be supplied in a form that is ready for use (e.g., a tablet or capsule) or in a form requiring, for example, reconstitution or dilution (e.g., a powder) prior to administration. When IDO inhibitors are in a form that needs to be reconstituted or diluted by a user, the kit may also include diluents (e.g., sterile water), buffers, pharmaceutically acceptable excipients, and the like, packaged with or separately of IDO inhibitors. When combination therapy is contemplated, the kit may contain the various agents separately or they may easily be combined in the kit. Each component of the kit can be included in an individual container, and all of the various containers can be in a single package. A kit of the present invention can be designed for the conditions necessary to properly maintain the components housed therein (e.g., refrigeration or freezing).

[00203] A kit may contain a label or packaging insert including identifying information regarding the components therein and instructions regarding their use (e.g., dosage parameters, clinical pharmacology of the active ingredient(s), including mechanism of action, pharmacokinetics and pharmacodynamics, adverse effects, contraindications, etc.). Labels or inserts may include manufacturer information such as batch numbers and expiration dates. Packaging labels or inserts can be, for example, integrated into the physical structure housing the components, contained separately in the physical structure, or attached to a component of the kit (for example, an ampoule, whole or vial).

[00204] Labels or inserts may additionally include, or be embedded in, a computer-readable medium, such as a disk (e.g., hard disk, card, memory disk), optical disks such as CD- or DVD-ROM/ RAM, DVD, MP3, magnetic tape, or electronic storage media such as RAM and ROM or hybrids of these such as magnetic/optical storage media, FLASH media or memory cards. In some embodiments, the actual instructions are not present in the kit, but means for obtaining the instructions from a remote source, e.g., via the internet, are provided.

EXPERIMENTAL

[00205] The following examples are presented in order to provide those skilled in the art with a complete exposition and description of how to prepare and use the present invention, and are not intended to limit the scope of what the inventors consider to be their invention, nor if are intended to represent that the experiments below have been performed or that they are all the experiments that can be performed. It should be understood that the exemplary descriptions written in the present tense were not necessarily carried out, but preferably the descriptions can be carried out to generate data and the like of a described nature in this regard. Efforts have been made to ensure accuracy with respect to the numbers used (e.g. quantities, temperature, etc.), but some deviations and experimental errors must be considered.

[00206] Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Celsius (°C), and pressure is at or near atmospheric. Standard abbreviations are used, including the following: wt = wild type; bp = base pair(s); kb = kilobase(s); nt = nucleotides(s); aa = amino acid(s); s or sec = second(s); minutes = minute(s); h or hr = hour(s); ng = nanogram; μg = microgram; mg = milligram; g = grass; kg = kilogram; dl or dL = deciliter; μl or μL = microliter; ml or mL = milliliter; l or L = liter; μM = micromolar; mM = millimolar; M = molar; kDa = kilodalton; i.m. = intramuscular(ly); i.p. = intraperitone- al(ly); SC or SQ = subcutaneously(ly); QD = daily; BID = twice daily; QW = weekly; QM = monthly; HPLC = high performance liquid chromatography; BW = body weight; U = unit; ns = not statistically significant; PBS = phosphate buffered saline; IHC = immunohistochemistry; DMEM = Dulbecco's Modification of Eagle's Medium; EDTA = ethylenediaminetetraacetic acid.

Materials and methods

[00207] The following general materials and methods were used, where indicated, or may be used in the Examples below:

[00208] Standard methods in molecular biology are described in the scientific literature (see, for example, Sambrook et al., Molecular Clo-ning, Third Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (2001); and Ausubel et al., Current Protocols in Molecular Biology, Vols. 1-4, John Wiley and Sons, Inc. New York, NY (2001), which describes cloning in bacterial cells and DNA mutagenesis (Volume 1), cloning in mammalian cells and yeast (Volume 2), protein expression and glycoconjugates (Volume 3), and bioinformatics (Volume 4)).

[00209] The scientific literature describes methods for protein purification, including immunoprecipitation, chromatography, electrophoresis, centrifugation, and crystallization, as well as chemical analysis, chemical modification, post-translational modification, production of fusion proteins, and protein glycosylation (see, for example, Coligan, et al., Current Protocols in Protein Science, Vols. 1-2, John Wiley and Sons, Inc., NY (2000)).

[00210] Software packages and databases for determining, for example, antigenic fragments, leader sequences, protein duplications, functional domains, glycosylation sites, and sequence alignments, are available (see, for example, GCG Wisconsin Package (Accelrys, Inc., San Diego, CA); and DECYPHER® (TimeLogic Corp., Crystal Bay, NV).

[00211] The literature is replete with assays and other experimental techniques that can serve as a basis for evaluating the compounds described here.

[00212] An IDO enzyme assay and cellular production of kynurenine (KYN) is described in Sarkar, S.A. et al., Diabetes, 56:72-79 (2007). Briefly, all therapeutic products can be purchased from Sigma-Aldrich (St. Louis, MO) unless otherwise specified. Groups of 1,000 human islets can be cultured for 24 hours in 1 ml of cytokine medium, recovered by centrifugation for 5 minutes at 800 x g, and sonicated in 150 μL of PBS containing a protease inhibitor cocktail (Set 2; Calbiochem, EMD Biosciences, San Diego, CA). The sonicate can be centrifuged for 10 minutes at 10,000 x g, and the supernatant can be assayed in triplicate by incubating a 40 μl sample with an equal value of 100 mmol/L potassium phosphate buffer, pH 6.5, containing 40 mmol/L. L of ascorbic acid (neutralized to pH 7.0), 100 μmol/L of methylene blue, 200 μg/mL of catalase, and 400 μmol/L of L-Trp for 30 minutes at 37 °C. The assay was terminated by adding 16 μL 30% (w/v) trichloroacetic acid (TCA) and also incubated at 60 °C for 15 minutes to hydrolyze N-formylkynurenine to KYN. The mixture can then be centrifuged at 12,000 rpm for 15 minutes, and KYN can be quantified by mixing equal volume of supernatant with 2% (w/v) Ehrlich reagent in glacial acetic acid in a 96-well microtiter plate and the absorption reading at 480 nm using L-KYN as standard. Protein in islet samples can be quantified by Bio-Rad protein assay at 595 nm. For detection of L-KYN in islet culture supernatants, proteins can be precipitated with 5% (w/v) TCA and centrifuged at 12,000 rpm for 15 minutes, and determination of KYN in the supernatant with Ehrlich reagent can be determined as described above. IL-4 (10 μg/mL; 500-2,000 units/mL) and 1-α-methyl Trp (1-MT; 40 μmol/L) can be added to the incubation media as indicated. This assay can also form the basis of a cell-based assay, and can be quantified using LCMS/MS as an alternative to UV/Vis detection.

[00213] Western Blot Analysis. Groups of 1,000 -1,200 islets incubated for 24 hours in Miami medium in the presence of cytokines can be harvested and sonicated in PBS as above, and 50 μg protein samples can be electrophoresed on SDS-PAGE gels COS7 cells (0, 6 x 10 cells/60 mm Petri dish) transfected with human IDO plasmid (3 μg) or empty vector cells can be used as negative and positive controls, respectively. Proteins can be transferred electrophoretically onto polyvinylidine fluoride membranes by semi-dry method and blocked for 1 hour with 5% (w/v) skimmed milk powder in Tris-buffered saline and 0.1% Tween and in then incubated overnight with mouse anti-human IDO antibody (1:500; Chemicon, Temecula, CA), phospho-STAT1α p91, and STAT1α p91 (1:500; Zymed, San Francisco, CA). Immunoreactive proteins can be visualized with ECL PLUS® Western Blotting Detection Reagent (Amersham BioSciences, Buckinghamshire, UK) after incubation for 1 hour with horseradish peroxidase-conjugated anti-mouse secondary antibody (Jackson Immunolabs , West Grove, PA).

[00214] Immunohistochemical detection of IDO. Islets can be fixed in 4% paraformaldehyde in PBS (Invitrogen) for 1 hour, immobilized in melted 10% porcine skin gelatin blocks (37°C), and embedded in optimal cutting temperature compound. Immunofluorescent staining on islet tissue can be performed on 7 μm sections that have been stained with elevated antibodies against pancreatic duodenal homeobox-1 (PDX1) and IDO. Antigen retrieval can be performed in a water bath for 30 minutes in a buffer containing 10 mmol/l Tris and 1 mmol/l EDTA (pH 9.0) at 97 °C. Sections can be blocked for 1 hour with 5% normal goat serum in PBS. Tissues can then be reacted with mouse monoclonal anti-human IDO antibodies (1:20; Chemicon) and goat polyclonal anti-human PDX1 antibody (1:2,000; which may be requested from Dr. Chris Wright, School of Medicine, Vanderbilt, TN) overnight at room temperature in a humidity chamber. Secondary anti-goat (Cy3-labeled) and anti-mouse (Cy2-labeled) antibodies can be purchased from Jackson Immunolabs and can be used at a concentration of 1:200. Nuclei can be stained with Hoechst 33258 (Molecular Probes, Eugene, OR). Images can be acquired by Intelligent Imaging System software from an Olympus 1X81 inverted motorized microscope equipped with Olympus DSU (spinning disk confocal) and Hamamatsu OR-CA IIER monochrome CCD camera.

[00215] Alternative means for evaluating the IDO inhibitors of the present invention are described in WO 2010/0233166 and are summarized here below.

[00216] Biochemical test. cDNA clones for both human and mouse IDO have been isolated and verified by sequencing and are commercially available. In order to prepare IDO for biochemical studies, C-terminal His-labeled IDO protein can be produced in E. coli using the IPTG-inducible pET5a vector system and isolated on a nickel column. The production of partially purified protein can be verified by gel electrophoresis and the concentration estimated by comparison to protein standards. For assay of IDO enzymatic activity, a 96-well plate spectrophotomeric assay for kynurenine production can be performed following published procedures (see, e.g., Littlejohn, T.K., et al., Prot. Exp. Purif., 19:22-29 (2000)). For analysis for IDO inhibitory activity, compounds can be evaluated at a single concentration of, for example, 200 μM against 50 ng of IDO enzyme in 100 μL reaction volumes with tryptophan added in increasing concentrations at, for example, 0. 2, 20, and 200 μM. Kynurenine production can be measured in 1 hour.

[00217] Cell-based assay. COS-1 cells can be transiently transfected with a CMV promoter-driven plasmid expressing IDO cDNA using Lipofectamine 2000 (Invitrogen) as recommended by the manufacturer. A complementary set of cells can be transiently transfected with plasmid expressing TDO. Forty-eight hours after transfection, cells can be portioned in a 96-well format at 6x104 cells per well. The next day, the wells can be washed and new medium (phenol-free) containing 20 μg/mL tryptophan can be added along with the inhibitor. The reaction can be stopped at 5 hours and the supernatant removed and spectrophotometrically assayed for kynurenine as previously described for the enzyme assay. To obtain initial confirmation of IDO activity, compounds can be evaluated at a single concentration of, for example, 100 μM. More extensive dose escalation profiles can be collected for selected compounds.

[00218] Pharmacodynamic and Pharmacokinetic Assessment. A pharmacodynamic assay can be based on measuring serum levels of both kynurenine and tryptophan, and calculating the kynurenine/tryptophan ratio provides an estimate of IDO activity that is independent of baseline tryptophan levels. Serum tryptophan and kynurenine levels can be determined by HPLC analysis, and serum compound levels can optionally also be determined in the same HPLC cycle.

[00219] Compounds can be initially evaluated by challenging mice with LPS and then subsequently administering a bolus dose of compound at the time when the serum kynurenine level reaches a plateau. When the kynurenine pool is rapidly changed with a serum half-life of less than 10 minutes, pre-existing kynurenine is not expected to unduly mask the impact that an IDO inhibitor has on kynurenine production. Each experiment may include mice not exposed to LPS (to determine baseline kynurenine levels with which to compare the other mice) and a set of LPS-exposed mice dosed with vehicle alone (to provide a positive control for IDO activation). . Each compound can initially be evaluated in mice in a single bolus dose i.p. high in the range of at least 100 mg/kg. Blood can be collected at defined time intervals (e.g., 50 μL sample at 5, 15, 30 min., 1, 2, 4, 6, 8, and 24 hr. after compound administration) for analysis of HPLC of kynurenine and tryptophan levels (pharmacodynamic analysis) as well as compound level (pharmacokinetic analysis). From the pharmacokinetic data, the peak serum concentration of the compound obtained can be determined, as well as the estimated clearance rate. By comparing the compound level in serum to the kynurenine/tryptophan ratio at various points, the effective IC50 for IDO inhibition in vivo can be approximately estimated. Compounds exhibiting efficacy can be evaluated to determine a maximum dose that achieves 100% IDO inhibition at peak concentration.

EXAMPLES

[00220] The invention is now described with reference to the following examples. These examples are provided for the purpose of illustration only and the invention should in no way be construed as being limited to these examples, but should preferably be construed to encompass any and all variations that become apparent as a result of the teaching provided here.

[00221] Abbreviations as used herein are defined as follows: "1 x" for once, "2 x" for twice, "3 x" for three times, "°C" for degrees Celsius, "eq" for equivalents, "g" for grams, "mg" for milligrams, "L" for liter or liters, "mL" for milliliter or milliliters, "μL" for microliter or microliters, "N" for normal, "M" for molar, "mmol" for millimol or millimoles, "min" for minute or minutes, "h" for hour or hours, "rt" for room temperature, "Tr" for retention time, "atm" for atmosphere, "psi" for pounds per square inch, "conc." for concentrated or concentrated, "aq" for "aqueous", "sat" or "sat'd" for saturated, "MW" for molecular weight, "mp" for melting point, "MS" or "Mass Spec" for spectrometry mass spectrometry, "ESI" for electrospray ionization mass spectroscopy, "HR" for high resolution, "HRMS" for high resolution mass spectrometry, "LCMS" for liquid chromatography mass spectrometry, "HPLC " for high pressure liquid chromatography, "RP HPLC" for reversed phase HPLC, "TLC" or "tlc" for thin layer chromatography, "NMR" for nuclear magnetic resonance spectroscopy, "nOe" for nuclear Overhauser effect spectroscopy , "1H" for proton, "δ" for delta, "s" for singlet, "d" for doublet, "t" for triplet, "q" for quartet, "m" for multiplet, "br" for broad, "Hz" for hertz, and "α", "β", "R", "S", "E", and "Z" are stereochemical designations familiar to one skilled in the art.

[00222] The compounds of the present invention can be prepared in various ways known to one skilled in the art of organic synthesis. The compounds of the present invention can be synthesized using the methods described below, together with synthetic methods known in the art of synthetic organic chemistry, or by variations thereon as appreciated by those skilled in the art. Preferred methods include, but are not limited to, those described below. Reactions are carried out in a solvent or solvent mixture appropriate to the reagents and materials used and suitable for the transformations being carried out. It will be understood by those skilled in the technique of organic synthesis that the functionality present in the molecule must be consistent with the proposed transformations. This will sometimes require judgment to modify the order of synthetic steps or to select one particular process scheme over another in order to obtain a desired compound of the invention.

[00223] The new compounds of this invention can be prepared using the reactions and techniques described in this section. Furthermore, in the description of the synthetic methods described below, it should be understood that all proposed reaction conditions, including choice of solvent, reaction atmosphere, reaction temperature, experiment duration, and preparation procedures, are chosen to be conditions patterns for that reaction, which should be easily recognized by someone skilled in the art. Restrictions on substituents that are compatible with the reaction conditions will be readily apparent to those skilled in the art and alternative methods should then be used.

SYNTHESIS PREPARATION METHODS

[00224] The compounds of the present invention can be prepared by methods such as those illustrated in the following schemes using chemical transformations known to those skilled in the art. Solvents, temperatures, pressures, and other reaction conditions can be easily selected by one skilled in the art. Starting materials are commercially available or readily prepared by one skilled in the art. These schemes are illustrative and are not intended to limit the possible techniques that one skilled in the art can use to manufacture the compounds described herein. Different methods may be apparent to those skilled in the art. Additionally, the various steps in the synthesis can be performed in an alternating sequence or order to provide the desired compound(s). Furthermore, the representation of reactions in these Schemes as discrete steps does not prevent them from being carried out in series, or by multiple telescopic steps in the same reaction vessel, or by carrying out multiple steps without purification or characterization of the intermediate(s). Furthermore, most of the compounds prepared by the methods below can also be modified using conventional chemistry well known to those skilled in the art. All documents cited here are incorporated herein by reference in their entirety.

[00225] References to many of these chemical transformations employed here can be found in Smith, M.B. et al., March's Advanced Organic Chemistry Reactions, Mechanisms, and Structure, Fifth Edition, Wiley-Interscience, New York (2001), or others standard texts on the topic of synthetic organic chemistry. Certain transformations may require reactive functional groups to be masked by protecting group(s). A convenient reference that provides conditions for introduction, removal, and relative susceptibility to reaction conditions of these groups is Greene, T.W. et al., Protective Groups in Organic Synthesis, Third Edition, Wiley-Interscience, New York (1999).

[00226] Referring to Scheme I, treatment of cyclohexanone II under standard Horner-Wadsworth Emmons conditions with phosphonate III will provide the corresponding unsaturated ester. Catalytic hydrogenation with, for example, Pd/C and hydrogen gas and subsequent ketal hydrolysis under acidic conditions provides the attached cycloalkanone of general structure IV. Treatment of compound IV with triflic anhydride and an organic base such as 2,6-lutidine will provide a vinyl triflate of general structure V. Coupling of V with arylboronic acids or EB(OR)2 esters, preferably under Suzuki conditions (see Kotha, S. et al., Tetrahedron, 58:9633-9695 (2002)) provides cycloalkenes of general structure VI. Typically, this reaction is carried out by heating the halide and ester or boronic acid to about 90 to about 98°C with the base such as aqueous tribasic sodium or potassium phosphate or potassium carbonate in a solvent such as dioxane, DMF. , THF, or NMP using a catalyst such as tetra-cis(triphenylphosphine)palladium or Cl2Pd(dppf). Many variations on this reaction involving the use of different temperatures, solvents, bases, anhydrous conditions, boronate derivatives, and halide substitutes such as triflates are known to those skilled in the art of organic/medical chemistry. Mild conditions have been reported for the coupling of sensitive boronic acid derivatives. See, Kinzel, T. et al., J. Am. Chem. Soc., 132(40):14073-14075 (2010). Saturation of the olefin in VII can be accomplished by treatment with Pd/C in a hydrogen atmosphere to provide a compound of general structure VII as a mixture of cis and trans isomers around the carbocycle. Further substitution of the ester can be accomplished by treatment with a strong base, such as LDA or LiHMDS, followed by addition of an electrophile R4-X where X is Br or I, to provide compounds of general structure VIII after basic hydrolysis with such a base. like LiOH. Coupling VIII with amines of general structure IX under standard conditions, well known to those skilled in the art, will provide compounds of general structure I. Scheme 1

[00227] As depicted in Scheme 2, olefin VI can be hydroborated by treatment with a borane, such as catechol borane, followed by standard oxidative preparation with hydrogen peroxide to provide a hydroxylated compound of general structure X, most likely as a mixture of isomers. Compound X can then be converted to a compound of general structure I by the methods described in Scheme 1. Scheme 2

[00228] In scheme 3, N-alkylation of a protected piperidinone of general structure XI can be performed by treatment with a haloacetate of general structure III (X=Br, Cl), subsequent acidic hydrolysis of the ketal will provide a keto ester of general structure XII. Formation of vinyl triflate, as previously described, will provide a compound of general structure XIII. Treatment of vinyl triflate with a diborane, such as bis-pinnacolatoborane, in the presence of a Pd(0) source, such as (PPh3)4Pd, will provide a vinyl boronic ester of general structure XIV. Suzuki coupling of aryl halites, EX, where X=Br, I, Cl, OTf, under standard conditions previously described, will provide an unsaturated compound of general structure XV. Compounds of general structure XV can be converted to compounds of general structure I by methods previously described herein. In another embodiment, compounds of general structure XV may first be treated with a borane, such as catechol borane, followed by oxidative preparation with hydrogen peroxide, to provide compounds of general structure XVI, which can be converted into compounds of general structure I by methods already described. Scheme 3

[00229] Scheme 4 describes the formation of amide through an anhydride of general structure XVIII. Treatment of an acid of general structure XVII with a chlorinating reagent, such as oxalyl chloride or thionyl chloride, will provide the desired acyl chloride of general structure XVIII. Compounds of general structure XVIII can be converted to amides of general structure I by treatment with an amine of general structure IX and an organic base, such as diisopropylethylamine. Scheme 4

[00230] Scheme 5 describes how a selective alkylation can be carried out to install R3 in the alpha position of amides that are of general structure I. Under standard conditions, mixed anhydrides of general structure XIX can be treated with chiral auxiliaries derived from oxazolidinone metal plate of general structure XX to provide an imide of general structure XXI. Evan's chiral auxiliaries are well known to those skilled in the art. Selective alkylation of XXI is provided by treatment with a strong base, such as NaHMDS, and an electrophile R3-X to provide a compound of general structure XXII with high diastereoselectivity for incorporation of R3. Hydrolysis of imide XXII can be carried out by standard methods such as a strong base and hydrogen peroxide (see Evans et al., Tetrahedron Lett., 28:6141-6144 (1987)) to provide a compound of general structure XXIII. Acid XXIII can be converted into a compound of general structure I by methods already described here. Scheme 5

[00231] In scheme 6, a ketone of general structure XXIV, which can be prepared by the methods described in schemes 1 and 3, can be treated under strongly reduced conditions with a borohydride such as sodium borohydride, to provide an alcohol of general structure XXV. Alcohol can be treated with a strong base in the presence of halo-substituted heteroaromatics to provide an ether of general structure XXVI. Alternatively, alcohol XXV can be treated under standard Mitsunobu DIAD and triphenylphosphine conditions to provide ethers of general structure XXVI, which can be converted to compounds of general structure I by methods already described herein. Scheme 6

[00232] Scheme 7 demonstrates how a ketone of general structure XXIV can be converted into an amine of general structure XXVII through reductive amination. This can be accomplished first by sequential treatment with an amine followed by a reducing agent such as sodium borohydride. The amine in XXVII can be attached by EX where X=Cl, Br or I through thermal conditions, such as heating in a solvent such as DMF, or through palladium-catalyzed coupling, such as a Buchwald coupling, to provide an amine of general structure XXVIII. The ester of general structure XXVIII can then be converted to a compound of general structure I by methods described herein. Scheme 7

[00233] As shown in scheme 8, a ketone of general structure XXIX can be treated with a haloacetate of general structure III in the presence of activated zinc metal to provide a tertiary alcohol of general structure XXX. The ester of general structure XXX can then be converted into a compound of general structure I using methods already described here. Scheme 8

[00234] As shown in scheme 9, a ketone of general structure XXIV can be treated with a metallated species EM, where M = Li, Na or K, produced by treating an aryl halide with, for example, an alkyl lithium, such as tert-butyllithium, to produce a tertiary alcohol of general structure XXXI. The ester of general structure XXXI can be converted into a compound of general structure I using methods already described here. Scheme 9

[00235] As shown in scheme 10, a monoprotected diamine of general structure XXXII can be converted into a compound of general structure XXXIII by methods described in scheme 3. Treatment of an amine of general structure XXXIII with EX, where X = Cl, Br, I, and a palladium catalyst, such as Pd(Ph3P)4, will provide a compound of general structure XXXIV. Alternatively, an amine of general structure XXXIII can be treated with a compound ECH2X under basic conditions sufficient for N-alkylation to provide a compound of general structure XXXIV. An ester of general structure XXXIV can then be converted into a compound of general structure I by methods already described herein. Scheme 10

HPLC/MS AND PREPARATORY/ANALYTICAL HPLC METHODS USED IN THE CHARACTERIZATION OR PURIFICATION OF EXAMPLES

[00236] Analytical and preparatory HPLC/MS was performed using the following methods:

[00237] Method A: Waters Acquity SDS using the following methods: Linear gradient from 2% to 98% Solvent B for 1.7 min; UV visualization at 220 nm; Column: BEH C18 2.1 mm x 50 mm; 1.7 μm particle (Heated to Temp. 50 °C); Flow rate: 0.8 ml/min; Mobile phase A: 100% water, 0.05% TFA; Mobile phase B: 100% acetonitrile, 0.05% TFA.

[00238] Method B: Column: Waters Acquity UPLC BEH C18, 2.1 x 50 mm, 1.7-μm particles; Mobile phase A: 5:95 acetonitrile:water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile:water with 10 mM ammonium acetate; Temperature: 50°C; Gradient: 0-100% B for 3 minutes, then a 0.75 minute hold at 100% B; Flow: 1.00 mL/min; Detection: UV at 220 nm.

[00239] Method C: Berger SFC MGII, Column: IC 25 x 3 cm ID, 5μm Flow rate: 85.0 mL/min, Mobile phase: 70/30 CO2/MeOH, Detector wavelength: 220 nm

[00240] Method D: Berger SFC analytical SFC, Column: Chiral IC 250 x 4.6 mm ID, 5μm, Flow rate: 2.0 mL/min, Mobile phase: 70/30 CO2/MeOH

[00241] Method E: Berger SFC MGII, Column: Chiral OJ-H 25 x 3 cm ID, 5μm Flow rate: 85.0 mL/min, Mobile phase: 75/25 CO2/MeOH, Detector wavelength: 220nm

[00242] Method F: Aurora analytical SFC, Column: Chiral IC 250 x 4.6 mm ID, 5μm, Flow rate: 2.0 mL/min, Mobile phase: 70/30 CO2/MeOH

[00243] Method G: Berger SFC MGII, Column: Chiral AS 25 x 3 cm ID, 5μm Flow rate: 85.0 mL/min, Mobile phase: 87/13 CO2/MeOH, Detector wavelength: 220 nm

[00244] Method H: Aurora analytical SFC, Column: Chiral AS 250 x 4.6 mm ID, 5μm, Flow rate: 2.0 mL/min, Mobile phase: 85/15 CO2/MeOH

[00245] Method I: Berger SFC MGII, Column: Chiral AS 25 x 3 cm ID, 5μm Flow rate: 85.0 mL/min, Mobile phase: 90/10 CO2/MeOH w/ 0.1% DEA, Detector wavelength: 220 nm

[00246] Method J: Aurora analytical SFC, Column: Chiral AS 250 x 4.6 mm ID, 5μm, Flow rate: 2.0 mL/min, Mobile phase: 90/10 CO2/MeOH w/ 0.1% from DEA

[00247] Method K: Column: Waters Acquity UPLC BEH C18, 2.1 x 50 mm, 1.7 μm particles; Mobile phase A: 5:95 acetonitrile:water with 0.1% trifluoroacetic acid; Mobile phase B: 95:5 acetonitrile:water with 0.1% trifluoroacetic acid; Temperature: 50°C; Gradient: 0-100% B for 3 minutes, then a retention of 0.75 to 100% B; Flow: 1.0 mL/min; Detection: UV at 220 nm.

[00248] Method L: Chiral HPLC: IF-3 column, ID 4.6 mm x 150 mm, Flow rate: 1 mL/min, Mobile phase: 85% Heptanes/15% isopropanol.

[00249] Method M: Chiral HPLC: ID column CHIRALPAK®, Chiral Technologies, West Chester, PA, 5 μm, ID 4.6 mm x 250 mm, Flow rate: 20 mL/minutes, Mobile phase: 0.4% of Et2NH in acetonitrile.

NMR USED IN EXAMPLE CHARACTERIZATION

[00250] 1H NMR spectra (unless otherwise noted) were obtained with JEOL® or Bruker FOURIER® transform spectrometers operating at 400 MHz or 500 MHz.

[00251] Spectral data is reported as chemical shift (multiplicity, number of hydrogens, coupling constants in Hz) and is reported in ppm (δ units) with respect to an internal standard (tetramethyl silane = 0 ppm) for 1H spectra NMR, or are referenced to residual solvent peaks (2.49 ppm for CD3SOCD2H, 3.30 ppm for CD2HOD, 1.94 for CHD2CN, 7.26 ppm for CHCl3, 5.32 ppm for CDHCl2). Abbreviations used in describing NMR peaks: "a" = apparent, "br. s." = broad singlet GENERAL PROCEDURES General Procedure A: Preparation of Aryl Cyclohexenes via Suzuki Cross-Coupling Reaction

[00252] Ethyl 2-(4-(((trifluoromethyl)sulfonyl)oxy)cycloex-3-en-1-yl)acetate* (1.0 equiv.), boronic acid (1.2 equiv), Na2CO3 (2.5 eq.), KBr (1.1 eq.) in 1,4-dioxane/water (10:1 by volume, 0.25 M) Pd(PPh3)4 (5 mol %) was added. . The resulting reaction mixture was heated to 80-90°C for 16 hours, whereupon the crude reaction mixture was concentrated. The resulting solids were diluted with EtOAc and water and the layers were separated. The aqueous layer was extracted with EtOAc three times. The combined organic extracts were dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The crude reaction mixture was purified using silica gel chromatography to provide the desired product.

[00253] * Ethyl 2-(4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)acetate is a known compound that can be prepared from 1,4-dioxaspiro[4,5] commercially available decan-8-one using the procedures outlined in 1) Stocks, PA et al., Angew. Chem. Int. Ed., 46:6278-6283 (2007); 2) Barlind, JG et al., J. Med. Chem., 55:10610-10629 (2012). General Procedure B: Hydrogenation

[00254] Unsaturated starting material was dissolved in a solvent of choice (e.g., methanol, ethyl acetate, or acetic acid) to form a 0.1-0.3 M solution. The resulting solution was purged with nitrogen and 20% by weight of a catalyst (10% by weight of dry activated Pd/C, or 10% by weight of Degussa Pd/C, or 10% by weight of Pd(OH)2/C) was added to the solution to form a heterogeneous mixture. Hydrogen gas was bubbled through the solution until complete disappearance of the starting material determined by TLC, and/or LC-MS, and/or NMR. Upon completion, the reaction mixture was purged with nitrogen, filtered through CELITE®, and concentrated under reduced pressure. The final product was purified by flash chromatography. General Procedure E: Ester Hydrolysis

[00255] To a solution of ester (1.0 eq) in EtOH (1.0 M) was added an equal volume of aqueous LiOH solution (7.25 M). The reaction mixture was stirred vigorously, heated to 50 °C for 1 hour and then diluted with 50 mL of water and also heated to 50 °C for 5 hours. The reaction mixture was cooled with an ice bath and acidified (pH ~ 1) by slow addition of 3M HCl solution. EtOAc was added, the layers were separated, and the aqueous phase was extracted with EtOAc (3x). The combined organic extracts were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to provide the desired carboxylic acid which was used without further purification. General Procedure G: Reaction Between Esters and Anilines

[00256] To a solution of aniline (2.0 eq.) in THF (0.25 M) at 0 °C was added a solution of PrMgCl (2.0 eq., 2 M in THF). The resulting solution was warmed to room temperature and stirred for 5 minutes at which point the ester (1.0 eq.) was added. The resulting reaction mixture was stirred at room temperature for 8 hours and poured into water. Ethyl acetate was added and the layers were separated. The aqueous layer was extracted three times with ethyl acetate. The combined organic extracts were dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure. The crude reaction mixture was purified using silica gel chromatography to provide the desired product. General Procedure K: Preparation of Aryl Cyclohexenes via Suzuki Cross-Coupling Reaction

[00257] Ethyl 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)acetate* (1,1 eq), aryl halide (1.0 eq), and Cs2CO3 (2.2 eq), in 1,4-dioxane/water (10:1 by volume, 0.25 M) catalytic amount of PEPPSI-IPr was added (2% Em mol). The resulting reaction mixture was heated to 100°C for 2-12 hours, whereupon the crude reaction mixture was concentrated and loaded onto silica gel. The crude reaction mixture was purified using silica gel chromatography.

[00258] * Ethyl 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)acetate is a known compound that can be prepared from commercially available 1,4-dioxaspiro[4,5]decan-8-one using the procedures outlined in Barlind, JG et al., J. Med. Chem., 55:10610-10629 (2012). General Procedure L: Coupling Carboxylic Acid with Chiral Auxiliary

[00259] To an oven-dried round base flask (Fial #1) carboxylic acid (1.0 eq) was added as a mixture of diastereomers. The flask was evacuated and recharged with nitrogen and subsequently charged with THF (0.25 M) and triethylamine (2.0 eq). The resulting solution was cooled to -78°C before slow addition of pivaloyl chloride (1.25 eq) over 15 minutes. The reaction mixture was then stirred at 0°C for one hour.

[00260] To a separate oven-dried round base flask (Fial #2) was added chiral (R)- or (S)-4-Benzyl- or 4-phenyl-2-oxazolidinone (1.3 eq) and THF (0.25M). This solution was cooled to -78 °C before careful addition of n-BuLi (2.5 M in hexanes, 1.3 eq). This reaction mixture was stirred at -78°C for 15 minutes before being removed from the cold bath.

[00261] Vial #1 was then cooled again to -78 °C and the contents of Vial #2 were added to Vial #1 via cannula over the course of 15 minutes. After complete addition, the ice bath was removed and the reaction was stirred for 3 hours at room temperature. The reaction was quenched by addition of saturated ammonium chloride solution (100 mL) and subsequent extraction with ethyl acetate (100 mL x 3). The combined organics were washed with brine, dried over sodium sulfate, filtered and concentrated. The crude residue was purified using silica gel chromatography. General Procedure M: Alkylation of imides derived from Oxazalidinone

[00262] NaHMDS (2 M in THF, 1.2 eq) was added dropwise to a 0.2 M solution of the imide (1.0 eq) in anhydrous tetrahydrofuran at -50 ° C. The solution was stirred for 10 minutes at -50°C and then liquid alkyl halide was added dropwise. The reaction mixture was stirred for an additional 2 to 48 hours at -50 to -20 ° C and then quenched by adding saturated ammonium chloride solution while still cold. The reaction mixture was allowed to warm to room temperature and was extracted 3 times with ethyl acetate. The combined organic extracts were dried with MgSO4, filtered, concentrated under reduced pressure, and subjected to flash chromatography on silica gel. General Procedure N: Chiral Helper Cleavage

[00263] An imide derived from oxazalidinone (0.418 mmol, 1.0 eq), THF (0.25 M) and distilled water (1M) were added to a round-base flask. This solution was cooled to 0 °C before the slow addition of H2O2 (35 wt% in water, 4 eq) followed by the addition of LiOH (2.7 M in water, 1.6 eq). The reaction solution was allowed to warm to room temperature. Progress was followed by LC/MS and the reaction solution was carefully quenched at 0 °C by addition of saturated Na2SO3 as the starting material was consumed. The pH was adjusted to ~5-6 with 1N HCl and then the mixture was extracted with EtOAc and methylene chloride. The combined organics were dried over sodium sulfate, filtered, and concentrated. The crude product was purified using silica gel chromatography. General Procedure O: Coupling of Carboxylic Acids and Anilines

[00264] Propylphosphonic anhydride (1.5 eq, 50% by weight of solution in ethyl acetate) was added to the solution of carboxylic acid (1 eq) and pyridine (3 eq) in ethyl acetate (0.1 M) at room temperature. The reaction mixture was stirred for 5 minutes, and then aniline (1.5 eq) was added. The reaction was stirred at room temperature until complete acid completion, which was determined by TLC and/or LC-MS. The reaction mixture was poured into water, 1M NaOH (10 eq) was added, and the aqueous layer was extracted with ethyl acetate three times. The combined organic extracts were dried with MgSO4 and concentrated in vacuo. The crude residue was purified by silica gel column chromatography. Examples 1 and 2 N-(4-chlorophenyl)-2-(trans -4-(quinolin-4-yl)cyclohexyl)acetamide N-(4-chlorophenyl)-2-(4-(cis-quinolin-4-yl)cyclohexyl)acetamide Examples 1 and 2: N -(4-chlorophenyl)-2-( trans -4-(quinolin-4-yl)cyclohexyl)acetamide and N-(4-chlorophenyl)-2-( cis -4-(quinolin-4 - il)cyclohexyl)acetamide

[00265] Examples 1 and 2 were prepared using General Procedures A, B, and G. General Procedure A employed 7.91 g (25 mmol) of 2-(4-(((trifluoromethyl)sulfonyl)oxy) cycloex- ethyl 3-en-1-yl)acetate and 4.56 g (26 mmol) of quinoline-4-boronic acid. General Procedure B employed 20 wt% dry Pd/C (10 wt%) and methanol as a solvent. General Procedure G employed 100 mg of ethyl 2-(4-(quinolin-4-yl)cyclohexyl)acetate (mixture of diastereomers) and 87 mg of 4-chloroaniline. Purified using silica gel chromatography (0% to 60% ethyl acetate in toluene) to provide Example 1 (trans-diastereomer) as the first eluting isomer. 1H trans-isomer NMR (400 MHz; CDCl3): δ 8.85 (d, J = 4.6 Hz, 1H), 8.04-8.14 (m, 2H), 7.67-7.72 (m, 1H), 7.53-7.59 (m, 1H), 7.47-7.53 (m, 2H), 7.27-7.31 (m, 3H), 3.27-3 .37 (m, 1H), 2.34 (d, J = 6.6 Hz, 2H), 2.03-2.11 (m, 5H), 1.61-1.73 (m, 2H), 1.31-1.44 (m, 2H) ppm, m/z 379.2 (M+H+).

[00266] Another elution from the column provided Example 2 (cis-diastereomer) as the second elution isomer. 1H cis-isomer NMR (400 MHz; CDCl3): δ 8.84 (d, J = 4.6 Hz, 1H), 8.04-8.14 (m, 2H), 7.67-7.73 (m, 1H), 7.48-7.60 (m, 3H), 7.25-7.31 (m, 3H), 3.36-3.46 (m, 1H), 2.51-2 .60 (m, 3H), 1.68-1.96 (m, 8H) ppm, m/z 379.2 (M+H+). Examples 3 and 4 N-(4-cyanophenyl)-2-( trans -4-(quinolin-4-yl)cyclohexyl)acetamide hydrochloride N-(4-cyanophenyl)-2-( cis -4-(quinolin-4-yl)cyclohexyl)acetamide hydrochloride Examples 3 and 4: N-(4-cyanophenyl)-2-( trans -4-(quinolin-4-yl)cyclohexyl)acetamide hydrochloride and N-(4-cyanophenyl)-2-( cis -4- (quinolin-4-yl)cyclohexyl)acetamide

[00267] Examples 3 and 4 were prepared using General Procedures A, B, and G. General Procedure A employed 7.91 g (25 mmol) of 2-(4-(((trifluoromethyl)sulfonyl)oxy)cyclohex ethyl -3-en-1-yl)acetate and 4.56 g (26 mmol) of quinoline-4-boronic acid. General Procedure B employed 20 wt% dry Pd/C (10 wt%) and methanol as a solvent. General Procedure G employed 100 mg of ethyl 2-(4-(quinolin-4-yl)cyclohexyl)acetate (mixture of diastereomers) and 80 mg of 4-cyanoaniline. Purified using silica gel chromatography (0% to 60% ethyl acetate in toluene) to provide Example 3 (trans-diastereomer) as the first eluting isomer. Free base was converted to hydrochloride by mixing with excess 2M HCl in diethyl ether, removing volatiles, and drying under high vacuum. 1H trans-isomer NMR (400 MHz; CDCl3): δ 10.53 (s, 1H), 9.2 (d, J = 5.7 Hz, 1H), 8.60 (d, J = 8.4 Hz, 1H), 8.33 (d, J = 8.6 Hz, 1H), 8.09-8.15 (m, 1H), 7.91-8.00 (m, 2H), 7.79 -7.84 (m, 2H), 7.72-7.77 (m, 2H), 3.60-3.70 (m, 1H), 2.37 (d, J = 6.7 Hz, 2H ), 1.87-1.99 (m, 5H), 1.65-1.77 (m, 2H), 1.34-1.47 (m, 2H) ppm. m/z 370.2 (M+H+).

[00268] Another elution from the column provided Example 4 (cis-diastereomer) as the second elution isomer. Free base was converted to hydrochloride by mixing with excess 2M HCl in diethyl ether, removing volatiles, and drying without high vacuum. 1H cis-isomer NMR (400 MHz; DMSO-d6): δ 10.79 (s, 1H), 9.26 (d, J = 5.7 Hz, 1H), 8.59 (d, J = 8 .4 Hz, 1H), 8.36 (d, J = 8.2 Hz, 1H), 8.09-8.17 (m, 2H), 7.91-7.97 (m, 1H), 7 .83-7.87 (m, 2H), 7.72-7.77 (m, 2H), 3,653.75 (m, 1H), 2.66 (d, J= 7.8 Hz, 2H), 2.37-2.46 (m, 1H), 1.81-1.99 (m, 4H), 1.65-1.77 (m, 4H) ppm. m/z 370.2 (M+H+). Examples 5 and 6 N-(4-fluorophenyl)-2-( trans -4-(quinolin-4-yl)cyclohexyl)acetamide N-(4-fluorophenyl)-2-( cis -4-(quinolin-4-yl)cyclohexyl)acetamide Examples 5 and 6: N-(4-fluorophenyl)-2-( trans -4-(quinolin-4-yl)cyclohexyl)acetamide and N-(4-fluorophenyl)-2-( cis -4-(quinolin-4 - il)cyclohexyl)acetamide

[00269] Prepared using General Procedures A, B, and G. General Procedure A employed 7.91 g (25 mmol) of 2-(4- (((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1 -yl)ethyl acetate and 4.56 g (26 mmol) of quinoline-4-boronic acid. General Procedure B employed 20 wt% dry Pd/C (10 wt%) and methanol as a solvent. General Procedure G employed 100 mg of ethyl 2-(4-(quinolin-4-yl)cyclohexyl)acetate (mixture of diastereomers) and 76 mg of 4-fluoroaniline. Purified using silica gel chromatography (0% to 60% ethyl acetate in toluene) to provide Example 5 (trans-diastereomer) as the first eluting isomer. 1H trans-isomer NMR (400 MHz; CDCl3): δ 8.85 (d, J = 4.5 Hz, 1H), 8.05-8.15 (m, 2H), 7.67-7.73 (m, 1H), 7.48-7.58 (m, 4H), 7.27 (d, J=4.7Hz, 1H), 6.98-7.05 (m, 2H), 3.25 -3.36 (m, 1H), 2.34 (d, J = 6.6 Hz, 2H), 2.01-2.10 (m, 5H), 1.58-1.72 (m, 2H ), 1.29-1.42 (m, 2H) ppm. m/z 363.2 (M+H+).

[00270] Another elution from the column provided Example 6 (cis-diastereomer) as the second elution isomer. 1H NMR of cis-isomer (400 MHz; CDCl3): δ 8.84 (d, J = 4.6 Hz, 1H), 8.05-8.15 (m, 2H), 7.67-7.73 (m, 1H), 7.54-7.62 (m, 2H), 7.48-7.54 (m, 2H), 7.27 (d, J = 4.6 Hz, 1H), 6, 97-7.05 (m, 2H), 3.36-3.46 (m, 1H), 2.51-2.60 (m, 3H), 1.68-1.98 (m, 8H) ppm . m/z 379.2 (M+H+). Examples 7 and 8 N-(4-fluorophenyl)-2-( cis -4-(quinolin-3-yl)cyclohexyl)acetamide N-(4-fluorophenyl)-2-( trans -4-(quinolin-3-yl)cyclohexyl)acetamide hydrochloride Examples 7 and 8: N-(4-fluorophenyl)-2-( cis -4-(quinolin-3-yl)cyclohexyl)acetamide and N-(4-fluorophenyl)-2-( trans -4-(quinolin) hydrochloride -3-yl)cyclohexyl)acetamide

[00271] Prepared using General Procedures A, B, and G. General Procedure A employed 10.0 g (32 mmol) of 2-(4- (((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1 -yl)ethyl acetate and 6.02 g (35 mmol) of quinoline-3-boronic acid. General Procedure B employed 20 wt% dry Pd/C (10 wt%) and methanol as a solvent. General Procedure G employed 95 mg of ethyl 2-(4-(quinolin-3-yl)cyclohexyl)acetate (mixture of diastereomers) and 71 mg of 4-fluoroaniline. Purified using silica gel chromatography (0% to 60% ethyl acetate in hexane) to provide Example 7 (cis-diastereomer) as the first eluting isomer. 1H cis-isomer NMR (400 MHz; DMSO-d6): δ 8.87-8.89 (d, J = 2.2 Hz, 1H), 8.18 (d, J = 1.8 Hz, 1H ), 7.91-8.00 (m, 2H), 7.65-7.72 (m, 1H), 7.55-7.64 (m, 3H), 7.09-7.15 (m , 3H) 2.77-2.87 (m, 1H), 2.47 (m, J = 8.0 Hz, 2H), 2.26-2.36 (m, 1H), 1.79-1 .91 (m, 2H), 1.57-1.78 (m, 6H) ppm. m/z 379.2 (M+H+).

[00272] Another elution from the column provided Example 8 (trans-diastereomer) as the second elution isomer. Free base was converted to hydrochloride by mixing with excess 2M HCl in diethyl ether, removing volatiles, and drying without high vacuum. 1H trans-isomer NMR (400 MHz; CDCl3): δ 10.05 (s, 1H), 9.21 (d, J = 1.8 Hz, 1H), 8.89 (s, 1H), 8, 18-8.26 (m, 2H), 7.96-8.03 (m, 1H), 7.81-7.87 (m, 1H), 7.60-7.65 (m, 2H), 7.08-7.16 (m, 2H), 2.83-2.93 (m, 1H), 2.27 (d, J = 6.6 Hz, 2H), 1.84-2.02 ( m, 5H), 1.59-1.71 (m, 2H), 1.15-1.28 (m, 2H) ppm. m/z 379.2 (M+H+). Examples 9 and 10 N-(4-chlorophenyl)-2-( cis -4-(quinolin-3-yl)cyclohexyl)acetamide

[00273] N-(4-chlorophenyl)-2-(trans-4-(quinolin-3-yl)cyclohexyl)acetamide Examples 9 and 10: N-(4-chlorophenyl)-2-( cis -4-(quinolin-3-yl)cyclohexyl)acetamide and N-(4-chlorophenyl)-2-( trans -4-(quinolin-3 - il)cyclohexyl)acetamide

[00274] Prepared using General Procedures A, B, and G. General Procedure A employed 10.0 g (32 mmol) of 2-(4- (((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1 -yl)ethyl acetate and 6.02 g (35 mmol) of quinoline-3-boronic acid. General Procedure B employed 20 wt% dry Pd/C (10 wt%) and methanol as a solvent. General Procedure G employed 95 mg of ethyl 2-(4-(quinolin-3-yl)cyclohexyl)acetate (mixture of diastereomers) and 82 mg of 4-chloroaniline. Purified using silica gel chromatography (0% to 60% ethyl acetate in hexane) to provide Example 9 (cis-diastereomer) as the first eluting isomer. 1H cis-isomer NMR (400 MHz; CDCl3): δ 8.80 (d, J = 2.2 Hz, 1H), 8.06-8.11 (m, 1H), 7.74-7.82 (m, 3H), 7.66-7.72 (m, 1H), 7.51-7.59 (m, 3H), 7.267.30 (m, 2H), 2.79-2.89 (m , 1H), 2.43-2.47 (m, 3H), 1.60-1.90 (m, 8H) ppm. m/z 379.1 (M+H+).

[00275] Another elution from the column provided Example 10 (trans-diastereomer) as the second elution isomer. 1H trans-isomer NMR (400 MHz; CDCl3): δ 8.81 (d, J = 2.2 Hz, 1H), 8.04-8.09 (m, 1H), 7.90-7.93 (m, 1H), 7.76-7.80 (m, 1H), 7.63-7.68 (m, 1H), 7.47-7.55 (m, 3H), 7.26-7 .32 (m, 2H), 7.24 (bs, 1H), 2.66-2.76 (m, 1H), 2.32 (d, J = 6.7 Hz, 2H), 1.99- 2.09 (m, 5H), 1.57-1.72 (m, 2H), 1.21-1.34 (m, 2H) ppm. m/z 379.1 (M+H+). Examples 11 and 12 N-(4-cyanophenyl)-2-( cis -4-(quinolin-3-yl)cyclohexyl)acetamide N-(4-cyanophenyl)-2-(trans -4-(quinolin-3-yl)cyclohexyl)acetamide Examples 11 and 12: N-(4-cyanophenyl)-2-( cis -4-(quinolin-3-yl)cyclohexyl)acetamide and N-(4-cyanophenyl)-2-( trans -4-(quinolin-3 - il)cyclohexyl)acetamide

[00276] Prepared using General Procedures A, B, and G. General Procedure A employed 10.0 g (32 mmol) of triflate and 6.02 g (35 mmol) of quinoline-3-boronic acid. General Procedure B employed 20 wt% dry Pd/C (10 wt%) and methanol as a solvent. General Procedure G employed 65 mg of ethyl 2-(4-(quinolin-3-yl)cyclohexyl)acetate (mixture of diastereomers) and 52 mg of 4-cyanoaniline. Purified using silica gel chromatography (0% to 60% ethyl acetate in hexane) to provide Example 11 (cis-diastereomer) as the first eluting isomer. 1H cis-isomer NMR (400 MHz; CDCl3): δ 8.76 (d, J = 2.3 Hz, 1H), 8.58 (bs, 1H), 8.07-8.11 (m, 1H ), 7.69-7.79 (m, 4H), 7.64-7.67 (m, 1H), 7.57-7.63 (m, 3H), 2.75-2.85 (m , 1H), 2.45-2.50 (m, 3H), 1.45-1.85 (m, 8H) ppm. m/z 370.2 (M+H+).

[00277] Another elution from the column provided Example 12 (trans-diastereomer) as the second elution isomer. 1H trans-isomer NMR (400 MHz; CDCl3): δ 8.80 (d, J = 2.3 Hz, 1H), 8.05-8.09 (m, 1H), 7.90-7.93 (m, 1H), 7.76-7.80 (m, 1H), 7.60-7.73 (m, 5H), 7.50-7.56 (m, 1H), 7.45 (bs , 1H), 7.67-7.77 (m, 1H), 2.36 (d, J = 6.7 Hz, 2H), 1.99-2.10 (m, 5H), 1.59- 1.72 (m, 2H), 1.22-1.35 (m, 2H) ppm. m/z 370.2 (M+H+). Examples 13 (a) and (b) 2-(4-(cis-1H-Indazol-4-yl)cyclohexyl)-N-(4-cyanophenyl)acetamide (first elution isomer, relative stereochemistry not determined and arbitrarily specified) 2-(4-(trans-1H-Indazol-4-yl)cyclohexyl)-N-(4-cyanophenyl)acetamide (second elution isomer, relative stereochemistry not determined and arbitrarily specified) Example 13: 2-(4 -(1H-Indazol-4-yl)cyclohexyl)-N-(4-cyanophenyl)acetamide

[00278] Prepared by General Procedures A, B and G. General Procedure A used indazol-4-boronic acid and 2-(4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl )ethyl acetate with dimethoxyethane as solvent, Pd(dppf)Cl2 as catalyst, K2CO3 as base, and omitted KBr as an additive. General Procedure B used 10 wt% Pd(OH)2/C (10 wt%) as catalyst and methanol as solvent. General Procedure G used 4-cyanoaniline (2 eq.) and ethyl 2-(4-(1H-indazol-4-yl)cyclohexyl)acetate (mixture of diastereomers) (1 eq). Purification of the reaction mixture by silica gel chromatography produced both isomers of Example 13. Example 13(a) (First elution isomer, relative stereochemistry not determined and arbitrarily specified): 1H NMR (400 MHz, CDCl3) δ 8.15 (s, 1H), 7.55 - 7.40 (m, 2H), 7.40 - 7.20 (m, 4H), 7.14 (s, 1H), 7.08 - 7.01 (m , 1H), 3.21 - 3.00 (m, 1H), 2.59 - 2.44 (m, 3H), 1.98 - 1.72 (m, 8H) ppm.

[00279] Example 13(b) (Second elution isomer, relative stereochemistry not determined and arbitrarily specified): 1H NMR (400 MHz, CDCl3) δ 8.13 (s, 1H), 7.69 (d, J = 8.8 Hz, 2H), 7.63 (d, J = 8.8 Hz, 2H), 7.37 - 7.20 (m, 3H), 6.99 (t, J = 4.0 Hz , 1H), 3.06 - 2.88 (m, 14H), 2.37 (d, J = 6.7 Hz, 2H), 2.13 - 1.97 (m, 5H), 1.83 - 1.65 (m, 2H), 1.39 - 1.27 (m, 2H) ppm. Example 14 2-(4-(1H-Indazol-4-yl)cyclohexyl)-N-(4-chlorophenyl)acetamide (mixture of diastereomers) Example 14: 2-(4-(1H-Indazol-4-yl)cyclohexyl)-N-(4-chlorophenyl)acetamide

[00280] Prepared by General Procedures A, B and G. General Procedure A used indazol-4-boronic acid and 2-(4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl )ethyl acetate with dimethoxyethane as solvent, Pd(dppf)Cl2 as catalyst, K2CO3 as base, and omitted KBr as an additive. General Procedure B used 10 wt% Pd(OH)2/C (10 wt%) as catalyst and methanol as solvent. General Procedure G used 4-chloroaniline (2 eq) and ethyl 2-(4-(1H-indazol-4-yl)cyclohexyl)acetate (1 eq). Purification of the reaction mixture by silica gel chromatography yielded Example 14 as a mixture of cis- and trans-diastereomers. 1H NMR (400 MHz, CDCl3) δ 8.13 (s, 1H), 7.55 - 7.45 (m, 2H), 7.38 - 7.21 (m, 4H), 7.13 (s, 1H), 6.99 (t, J = 3.9 Hz, 1H), 3.06 - 2.90 (m, 1H), 2.33 (d, J = 6.7 Hz, 2H), 2, 13 - 1.96 (m, 5H), 1.82 - 1.64 (m, 2H), 1.38 - 1.28 (m, 2H) ppm. m/z 368.2 (M+H+). Example 16 N-(4-fluorophenyl)-2-(cis-4-(quinolin-4-yl)cyclohexyl)propanamide (simple diastereomer, relative stereochemistry not determined and arbitrarily specified) Intermediate 16A: Ethyl 2-(4-(quinolin-4-yl)cyclohexyl)acetate

[00281] Intermediate 16A was prepared with General Procedures A and B. General Procedure A used ethyl 2-(4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)acetate and acid quinoline-4-boronic. General Procedure B employed 20% by weight of dry Pd/C (10% by weight) and methanol as a solvent to provide the desired intermediate product 16A. Intermediate 16B: Ethyl 2-(4-(quinolin-4-yl)cyclohexyl)propanoate

[00282] To a solution of Intermediate 16A (630 mg, 2.15 mmol) in THF (10 mL) at 0 °C was added NaHMDS solution (4.3 mL, 4.3 mmol, 1M in THF). The resulting yellow solution was stirred at 0°C for 5 minutes and Mel (608 mg, 4.3 mmol) was added. The reaction mixture was stirred at 0 °C for 1 hour, under which acetic acid (200 μL) was added along with Et2O (10 mL). The reaction mixture was filtered through a silica plug eluting with additional Et2O (50 mL). The filtrate was concentrated and purified using silica gel chromatography (15% to 30% EtOAc in hexanes) to provide Intermediate 16B as an oil and 2:1 mixture of cis:trans diastereomers. Example 16: N-(4-fluorophenyl)-2-( cis -4-(quinolin-4-yl)cyclohexyl)propanamide

[00283] Prepared with General Procedure G using Intermediate 16B (78 mg, 0.25 mmol) and 4-fluoroaniline (56 mg, 0.5 mmol). Purified using silica gel chromatography (30% to 45% ethyl acetate in hexanes) to provide Example 16 as a white solid and single diastereomer (first eluting isomer, racemic, relative stereochemistry was not determined). 1H NMR (400 MHz; CDCl3): δ 8.83 (d, J = 4.6 Hz, 1H), 8.11 (dd, J = 8.4, 0.8 Hz, 1H), 8.05 ( dd, J = 8.4, 0.5 Hz, 1H), 7.69 (ddd, J = 8.4, 6.9, 1.4 Hz, 1H), 7.57-7.52 (m, 4H), 7.26 (s, 1H), 7.05-6.99 (m, 2H), 3.31-3.25 (m, 1H), 2.22-2.15 (m, 1H) , 2.11-1.98 (m, 4H), 1.90 (s, 1H), 1.82-1.73 (m, 1H), 1.60 (qd, J = 11.8, 2, 6 Hz, 2H), 1.44-1.27 (m, 5H) ppm. m/z 377.3 (M+H+). Examples 17 and 18 N-(4-chlorophenyl)-2-( trans -4-(quinolin-4-yl)cyclohexyl)propanamide N-(4-chlorophenyl)-2-( cis -4-(quinolin-4-yl)cyclohexyl)propanamide Examples 17 and 18: N-(4-chlorophenyl)-2-(trans-4-(quinolin-4-yl)cyclohexyl) propanamide and N-(4-chlorophenyl)-2-(cis-4-(quinolin-4 -yl)cyclohexyl)propanamide

[00284] Prepared with General Procedure G using Intermediate 16B (44 mg, 0.14 mmol) and 4-chloroaniline (36 mg, 0.28 mmol). Purified using silica gel chromatography (30% to 45% EtOAc in hexanes) to provide Example 17 (trans-diastereomer, racemic) as a white solid and the first eluting isomer. 1H NMR (400 MHz; CDCl3): δ 8.84 (d, J = 4.6 Hz, 1H), 8.11 (dd, J = 8.4, 1.0 Hz, 1H), 8.07- 8.05 (m, 1H), 7.69 (ddd, J = 8.4, 6.9, 1.4 Hz, 1H), 7.58-7.50 (m, 3H), 7.31- 7.25 (m, 4H), 3.34-3.26 (m, 1H), 2.22-2.15 (m, 1H), 2.141.98 (m, 5H), 1.83-1, 74 (m, 1H), 1.65-1.57 (m, 2H), 1.44-1.32 (m, 2H), 1.30 (d, J = 6.9 Hz, 3H) ppm. m/z 393.2 (M+H+).

[00285] Another elution from the column in the previous example provided Example 18 (cis-diastereomer, racemic) as a white solid and the second eluting isomer. 1H NMR (400 MHz; CDCl3): δ 8.79 (d, J = 4.6 Hz, 1H), 8.11 (dd, J = 8.4, 1.2 Hz, 1H), 8.07 ( d, J = 8.0 Hz, 1H), 8.03 (s, 1H), 7.71 (ddd, J = 8.3, 6.9, 1.3 Hz, 1H), 7.58 (ddd , J = 8.4, 6.9, 1.4 Hz, 1H), 7.55-7.52 (m, 2H), 7.28-7.23 (m, 3H), 3.50-3 .42 (m, 1H), 2.68-2.60 (m, 1H), 2.18-2.12 (m, 1H), 1.95-1.67 (m, 8H), 1.29 (d, J = 6.8 Hz, 3H) ppm. m/z 393.2 (M+H+). Example 19 (R)-N-(4-chlorophenyl)-2-( cis -4-(6-fluoroquinolin-4-yl)cyclohexyl)propanamide Example 19: (R)-N-(4-chlorophenyl)-2-( cis -4-(6-fluoroquinolin-4-yl)cyclohexyl)propanamide

[00286] Prepared using General Procedures K, B, E, L, M, N, and O. General Procedure L employed 2-(4-(6-fluoroquinolin-4-yl)-cyclohexyl)acetic acid (mixture of diastereomers), and (R)-2-phenyl-oxazolidinone. General Procedure M employed the product cis and io-domethane. The auxiliary was removed after General Procedure N and the desired product formed using General Procedure O with 4-chloroaniline. Purified using silica gel chromatography (0% to 100% ethyl acetate in hexanes) to give Example 19. cis-isomer 1H NMR (400 MHz; CDCl3): δ 9.14 (s, 1H), 8. 70 (d, J = 4.6 Hz, 1H), 8.06 (dd, J = 9.2 Hz, J = 5.6 Hz, 1H), 7.58-7.64 (m, 3H), 7.45 (ddd, J = 9.3 Hz, J = 7.8 Hz, J = 2.7 Hz, 1H), 7.19-7.24 (m, 2H), 7.15 (d, J = 4.6Hz, 1H), 3.16-3.26 (m, 1H), 2.59-2.69 (m, 1H), 2.08-2.16 (m, 1H), 1.66 -1.86 (m, 7H), 1.31-1.42 (m, 1H), 1.21 (d, J = 6.8Hz, 3H) ppm. m/z 411.2 (M+H)+. Example 20 (S)-N-(4-chlorophenyl)-2-( trans -4-(quinolin-3-yl)cyclohexyl)propanamide Example 20: (S)-N-(4-chlorophenyl)-2-(trans -4-(quinolin-3-yl)cyclohexyl)propanamide

[00287] Prepared using General Procedures K, B, E, L, M, N, and O. General Procedure L employed 2-(4-(quinolin-3-yl)-cyclohexyl)acetic acid (mixture of diastereomers) , and (S)-2-phenyloxazolidinone. General Procedure M employed a mixture of diastereomers and iodomethane. The auxiliary was removed after General Procedure N and the desired product formed using General Procedure O with 4-chloroaniline. Purified using silica gel chromatography (0% to 10% isopropanol in hexanes) to provide Example 20 (trans-isomer) as a second eluting isomer. 1H trans-isomer NMR (400 MHz; MeOH): δ 9.21 (d, J = 1.9 Hz, 1H), 9.10 (d, J = 1.9Hz, 1H), 8.30-8 .34 (m, 1H), 8.19-8.23 (m, 1H), 8.10-8.16 (m, 1H), 7.94-8.00 (m, 1H), 7.57 -7.63 (m, 2H), 7.29-7.34 (m, 2H), 3.00 (tt, J = 12.0Hz, J = 3.1Hz, 1H), 2.28-2 .38 (m, 1H), 2.07-2.22 (m, 3H), 1.9302.01 (m, 1H), 1.65-1.82 (3H), 1.22-1.46 (m, 5H) ppm. m/z 393.2 (M+H)+. Example 21 (R)-N-(4-cyanophenyl)-2-( cis -4-(7-fluoroquinolin-4-yl)cyclohexyl)butanamide Example 21: (R)-N-(4-cyanophenyl)-2-( cis -4-(7-fluoroquinolin-4-yl)cyclohexyl) butanamide

[00288] Prepared using General Procedures K, B, E, L, M, N, and O. General Procedure L employed 2-(4-(7-fluoroquinolin-4-yl)-cyclohexyl)acetic acid (mixture of diastereomers), and (R)-2-phenyl-oxazolidinone. General Procedure M employed the product cis- and io-doethane. The auxiliary was removed after General Procedure N and the desired product formed using General Procedure O with 4-cyanoaniline. Purified using silica gel chromatography (10% to 25% EtOAc in CH2Cl2) to give Example 21 (cis-isomer). 1H NMR (400 MHz, CDCl3) δ 8.78 (d, J = 4.6 Hz, 1H), 8.55 (s, 1H), 8.07 (dd, J = 9.4, 5.9 Hz , 1H), 7.80 - 7.67 (m, 3H), 7.60 - 7.53 (m, 2H), 7.37 (ddd, J = 9.4, 7.9, 2.7 Hz , 1H), 7.26 (d, J = 5.2 Hz, 1H), 3.44 (s, 1H), 2.51 (td, J = 10.4, 3.7 Hz, 1H), 2 .23 - 2.11 (m, 1H), 2.09 - 1.35 (m, 10H), 1.01 (t, J = 7.4 Hz, 3H) ppm. m/z 416.2 (M+H)+. Example 22 (R)-N-(4-chlorophenyl)-2-( cis -4-(7-fluoroquinolin-4-yl)cyclohexyl)butanamide Example 22: (R)-N-(4-chlorophenyl)-2-( cis -4-(7-fluoroquinolin-4-yl)cyclohexyl) butanamide

[00289] Prepared using General Procedures K, B, E, L, M, N, and O. General Procedure L employed 2-(4-(7-fluoroquinolin-4-yl)-cyclohexyl)acetic acid (mixture of diastereomers), and (R)-2-phenyl-oxazolidinone. General Procedure M employed the cis product and iodoethane. The auxiliary was removed after General Procedure N and the desired product formed using General Procedure O with 4-chloroaniline. Purified using silica gel chromatography (20% to 50% EtOAc in hexanes). The residue was also purified using silica gel chromatography (25% EtOAc in CH2Cl2) to provide the desired product. 1H NMR (400 MHz, CDCl3) δ 8.83 (d, J = 4.6 Hz, 1H), 8.07 (dd, J = 9.4, 5.9 Hz, 1H), 7.74 (dd , J = 10.0, 2.7 Hz, 1H), 7.60 - 7.44 (m, 3H), 7.41 - 7.24 (m, 4H), 3.54 - 3.41 (m , 1H), 2.42 (td, J = 10.7, 3.7 Hz, 1H), 2.19 (d, J = 16.6 Hz, 1H), 2.17 - 1.37 (m, 10H), 1.02 (t, J = 7.4 Hz, 3H) ppm. m/z 425.2 (M+H)+. Example 23 (R)-N-(4-chlorophenyl)-2-( cis -4-(quinolin-4-yl)cyclohexyl)propamide Example 23: (R)-N-(4-chlorophenyl)-2-( cis -4-(quinolin-4-yl)cyclohexyl)propamide

[00290] Prepared using General Procedures K, B, E, L, M, N, and O. General Procedure L employed 2-(4-(quinolin-4-yl)-cyclohexyl)acetic acid (mixture of diastereomers) , and (R)-2-phenyloxazolidinone. General Procedure M employed the product cis and io-domethane. The auxiliary was removed following General Procedure N and the desired product formed employing General Procedure O with 4-chloroaniline to give Example 23. The undesired minor isomer was removed via Method L: retention time of minor isomer, unwanted = 7.1 min, larger enantiomer retention time, desired = 8.0 min. 1H NMR of desired enantiomer (400 MHz; CDCl3): δ 8.84 (d, J = 4.6 Hz, 1H), 8.29 (s, 1H), 8.14 - 8.02 (m, 2H) , 7.70 (ddd, J = 8.3, 6.8, 1.3 Hz, 1H), 7.62 - 7.54 (m, 2H), 7.32 - 7.20 (m, 2H) , 3.60 - 3.26 (m, 1H), 2.71 - 2.48 (m, 1H), 2.16 - 2.08 (m, 1H), 1.96 - 1.66 (m, 9H), 1.63 - 1.45 (m, 1H), 1.24 (d, J = 6.8 Hz, 3H) ppm. m/z 393.2 (M+H)+. Example 24 (R)-N-(4-cyanophenyl)-2-( cis -4-(quinolin-4-yl)cyclohexyl)propamide Example 24: (R)-N-(4-cyanophenyl)-2-( cis -4-(quinolin-4-yl)cyclohexyl)propamide

[00291] Prepared using General Procedures K, B, E, L, M, N, and O. General Procedure L employed 2-(4-(quinolin-4-yl)-cyclohexyl)acetic acid (mixture of diastereomers) , and (R)-2-phenyloxazolidinone. General Procedure M employed the product cis and io-domethane. The auxiliary was removed after General Procedure N and General Procedure O with 4-cyanoaniline provided Example 24. 1H NMR (400 MHz; CDCl3): δ 8.89 (d, J = 4.6 Hz, 1H), 8.41 (s, 1H), 8.11 (d, J = 7.6 Hz, 1H), 8.06 (d, J = 8.2 Hz, 1H), 7.78 (d, J = 8 .8 Hz, 2H), 7.74 - 7.67 (m, 1H), 7.64 - 7.54 (m, 3H), 7.33 (d, J = 4.6 Hz, 1H), 3 .54 - 3.37 (m, 1H), 2.67 - 2.53 (m, 1H), 2.16 - 2.09 (m, 1H), 2.00 - 1.72 (m, 7H) , 1.57 - 1.45 (m, 1H), 1.26 (d, J = 6.8 Hz, 3H) ppm. m/z 384.2 (M+H)+. Example 25 (R) - N-(4-chlorophenyl)-2-( cis -4-(quinolin-4-yl)cyclohexyl)butanamide Example 25: (R) - N-(4-chlorophenyl)-2-( cis -4-(quinolin-4-yl)cyclohexyl)butanamide

[00292] Prepared using General Procedures K, B, E, L, M, N, and O. General Procedure L employed 2-(4-(quinolin-4-yl)-cyclohexyl)acetic acid (mixture of diastereomers) and (R)-4-phenyl-2-oxazolidinone. General Procedure M used the cis product of procedure L and iodoethane. The auxiliary was removed after General Procedure N and the desired product formed using General Procedure O with 4-chloroaniline. The compound was purified using silica gel chromatography (10% to 100% EtOAc in methylene chloride) to provide Example 25 as a white solid. 1H NMR (400 MHz; CDCl3): δ 8.83 (d, J = 4.6 Hz, 1H), 8.15 - 7.95 (m, 3H), 7.69 (dd, J = 8.3 , 7.0 Hz, 1H), 7.63 - 7.49 (m, 3H), 7.31 - 7.22 (m, 3H), 3.47 (s, 1H), 2.43 (td, J = 10.4, 3.7 Hz, 1H), 2.16 - 2.13 (m, 1H), 1.98 - 1.52 (m, 11H), 1.01 (t, J = 7, 3Hz, 3H) ppm. m/z 406.2 (M+H+). Example 26 (S)-N-(4-chlorophenyl)-2-(trans-4-(quinolin-4-yl)cylohexyl)butanamide Example 26: (S)-N—(4-ç^rophenylj-2-(trans-4-(quinoin-4zyl)cyclohexyl)butanamide

[00293] Prepared using General Procedures K, B, E, L, M, N, and O. General Procedure L employed 2-(4-(quinolin-4-yl)-celiclohexyl)acetic acid (mixture of diastereomers) , and (S)-2-phenyloxazolidinone, General Procedure M employed iodoethane and the trans product from Procedure L. The auxiliary was removed after General Procedure N and the desired product formed using General Procedure O with 4-chloroaniline. The compound was purified using silica gel chromatography (10% to 100% EtOAc in methylene chloride) to provide Example 26 as a white solid. 1H NMR (400 MHz; CDCl3): δ 8.84 (d, J = 4.6 Hz, 1H), 8.11 (dd, J = 8.5, 1.1 Hz, 1H), 8.05 ( dd, J = 8.4, 0.6 Hz, 1H), 7.69 (ddd, J = 8.4, 6.8, 1.4 Hz, 1H), 7.59 - 7.46 (m, 3H), 7.34 - 7.29 (m, 2H), 7.26 - 7.25 (m, 2H), 3.30 (t, J = 11.7 Hz, 1H), 2.25 - 1 .88 (m, 4H), 1.85 - 1.69 (m, 3H), 1.70 - 1.50 (m, 3H), 1.50 - 1.27 (m, 2H), 0.99 (t, J = 7.4 Hz, 3H) ppm. m/z 406.2 (M+H+). Example 27 N-(4-chlorophenyl)-2-(cis-4-(quinolin-4-yloxy)cyclohexyl)propanamide (enantiomerically pure, absolute stereochemistry not determined) Intermediate 27A: 2-(1,4-dioxaspiro[4,5]decan-8-ylidene)propanoate

[00294] A suspension of 60% NaH (12.8 g, 385 mmol) in THF (500 mL) was cooled to 0 °C and triethyl-2-phosphono propionate (91.6 g, 385 mmol) was added for 30 minutes. The reaction mixture was allowed to warm to room temperature and the cloudy suspension gradually became a yellow solution. After warming to room temperature, the reaction mixture was cooled again to 0 °C. A solution of 1,4-cyclohexanedione monoethylene acetal (50 g, 320 mmol) was added dropwise via an addition funnel. After the addition was complete, the reaction mixture was allowed to warm to room temperature and stirred for 1 hour. The mixture was concentrated under reduced pressure and the yellow residue was diluted with saturated aqueous NaHCO3 (500 mL) and EtOAc (1000 mL). The organic layer was separated and the aqueous layer was extracted with EtOAc (500 mL x 2). The combined organic extracts were dried over anhydrous Na2SO4, filtered and concentrated under reduced pressure to provide Intermediate 27A (93.8 g), which was used in the next step without further purification. Intermediate 27B: Racemic ethyl 2-(1,4-dioxaspiro[4,5]decan-8-yl)propanoate

[00295] Intermediate 27A (93.8 g, 390.3 mmol) was dissolved in EtOH (1000 mL) and 10% palladium on carbon (9.4 g) was added. An H2(g) flask with a needle was sprayed through the black suspension, after which the flask was refilled and the vessel kept under an H2(g) atmosphere overnight. The mixture was filtered through CELITE®545 and the filter cake was rinsed with EtOH. The filtrate was concentrated under reduced pressure to provide Intermediate 27B (92.5 g), which was used in the next step without further purification. Intermediate 27C: Racemic ethyl 2-(4-oxocyclohexyl)propanoate

[00296] A racemic mixture of Intermediate 27B (17.1 g, 70.6 mmol) was dissolved in acetone (250 mL). Then, 1N aqueous HCl (62 mL) was added and the mixture was stirred at room temperature for 24 hours. The mixture was concentrated under reduced pressure and extracted with EtOAc (200 mL x 3). The combined organic extracts were washed with 1N aqueous NaOH (50 mL), dried over anhydrous MgSO4, filtered and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (0 - 20% EtOAc in petroleum ether) to provide Intermediate 27C (12.8 g, 91.5% yield). Intermediate 27D: ethyl-2-(trans-4-hydroxycyclohexyl)propanoate

[00297] To a solution of racemic Intermediate 27C (5.94 g, 30 mmol) in MeOH (100 mL) at 0 °C was added NaBH4 (1.67 g, 45 mmol) in portions. The mixture was warmed to room temperature and monitored by TLC. When the starting material was consumed, the mixture was quenched with 1 M HCl and extracted with CH2Cl2 (2 x 100 mL). The combined organic extracts were dried over MgSO4, filtered and concentrated under reduced pressure. The resulting pale yellow oil was purified by silica gel chromatography (25% to 40% EtOAc in hexanes) to provide the trans-isomer, Intermediate 27D as the second eluting isomer. Intermediate 27E. Ethyl-2-( cis -4-(quinolin-4-yloxy)cyclohexyl)propanoate:

[00298] To a solution of racemic Intermediate 27D (690 mg, 3.45 mmol), quinolin-4-ol (600 mg, 4.13 mmol) and triphenylphosphine (2.69 g, 10.3 mmol) in THF ( 11.5 mL) at 0 °C DEAD (0.921 mL, 5.87 mmol) was added dropwise. The mixture was warmed to room temperature and stirred at room temperature for 18 hours. The mixture was diluted with 1M NaOH and extracted with EtOAc (3x). The combined organic layers were washed with brine, dried over MgSO4, filtered and concentrated under reduced pressure. The residue was purified using silica gel chromatography (10% to 100% EtOAc in hexanes) to provide Intermediate 27E. Example 27: N-(4-chlorophenyl)-2-( cis -4-(quinolin-4-yloxy)cyclohexyl)propanamide

[00299] Example 27 was prepared with General Procedure G using Intermediate 27E and 4-chloroaniline. The racemic mixture was resolved using Method M to provide Example 27 as a single enantiomer (absolute stereochemistry not determined, second elution enantiomer). 1H NMR (400 MHz, CDCl3) δ 8.61 (d, J = 5.3 Hz, 1H), 8.27 (s, 1H), 8.14 (d, J = 8.4 Hz, 1H), 7.93 (d, J = 8.4 Hz, 1H), 7.61 (t, J = 7.6 Hz, 1H), 7.49 (d, J = 8.7 Hz, 2H), 7, 42 (t, J = 7.6 Hz, 1H), 7.31 - 7.13 (m, 2H), 6.62 (d, J = 5.3 Hz, 1H), 4.44 - 4.20 (m, 1H), 2.27 - 2.01 (m, 3H), 1.99 - 1.78 (m, 2H), 1.72 - 1.58 (m, 1H), 1.51 (dd , J = 24.1, 12.8 Hz, 2H), 1.24 - 0.96 (m, 5H). m/z 409.2 (M+H)+. Example 30 N-(4-Chlorophenyl)-2-(cis-1-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetamide (simple diastereomer, relative stereochemistry not determined and arbitrarily specified) Intermediate 30A: 4-(1,4-dioxaspiro[4,5]dec-7-en-8-yl)quinoline

[00300] To a 20 mL reaction vial containing 4,4,5,5-tetramethyl-2-(1,4-dioxaspiro[4,5]dec-7-en-8-yl)-1,3, 2-dioxaborolane (See General Procedure K for preparation reference) commercially available, CAS# [680596-79-6]) (1.0 g, 3.8 mmol, 1.0 equiv.), 4-bromoquinoline (0 .86 g, 4.1 mmol, 1.1 equiv.), and Pd(dppf)Cl2 (0.154 g, 0.188 mmol, 0.05 equiv.) were added dioxane (12 mL), water (1.2 mL) , and NEt3 (1.0 mL, 7.5 mmol, 2.0 equiv.). The flask was flooded with argon and placed in a preheated block at 100 °C for 15 hours. The crude residue was diluted with EtOAc (100 mL), filtered through a pad of CELITE®, and concentrated. The crude residue was purified by silica gel chromatography (0% to 50% EtOAc in hexanes) to provide Intermediate 30A (1.0 g, 99% yield). Intermediate 30B: 4-(quinolin-4-yl)cyclohexan-1-one

[00301] Intermediate 30A (3.25 g, 12.3 mmol, 1.0 equiv.) was dissolved in MeOH (100 mL) and sprayed with argon for 1 hour before adding Pd/C (2.54 g , 2.4 mmol, 0.2 equiv.). The reaction solution was sprayed with H2 (g) and placed under positive pressure with an H2 balloon. The reaction was stirred under H2 for 14 hours and filtered through CELITE®, rinsed with 100 mL of MeOH. The solution was concentrated to provide a light yellow solid oil mixture. The crude mixture was directly diluted with 3 M HCl (100 mL) and acetone (100 mL). The solution was stirred at room temperature for 2 hours and monitored by TLC. The reaction was basified with 1N Na-OH and extracted with EtOAc (2 x 100 mL), washed with brine, dried (MgSO4), filtered, and concentrated. The crude residue was purified by silica gel chromatography (0% to 50% EtOAc in hexanes) to provide Intermediate 30B (1.25 g, 45% production over 2 steps). Intermediate 30Ca: 2-(cis -1-hydroxy-4-(quinolin-4-yl)cyclohexyl)ethyl acetate Intermediate 30Cb: 2-(trans-1-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetate ethyl

[00302] To a 20 mL reaction vial containing Intermediate 30B (0.99 g, 4.4 mmol, 1.0 equiv.) and Zn0 (350 mg, 5.3 mol, 1.2 equiv.) under argon anhydrous THF (8 mL) was added. A solution of ethyl bromoacetate (0.54 mL, 4.8 mol, 1.1 equiv.) in THF (4.0 mL) was added via syringe followed by an iodine crystal. The reaction mixture was placed in a preheated block at 80 °C for 5 hours, after which time an additional 0.5 equiv. of ethyl bromoacetate (0.24 mL, 2.2 mmol) and Zn0 (150 mg, 2.2 mmol) were added. The reaction continued stirring at 80°C for an additional 2 hours. The mixture was diluted with EtOAc and filtered through a CELITE® pad. The solution was concentrated and purified by silica gel chromatography (0% to 100% EtOAc in hexanes) to provide Intermediate 30Ca as the trans and cis isomers of Intermediate 30Cb as yellow solids. Example 30: N-(4-Chlorophenyl)-2-( cis -1-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetamide

[00303] To a reaction vial containing 4-chloroaniline (56 mg, 0.44 mmol, 2.0 equiv.) iPrMgCl (2.0 M in THF, 0.22 mL, 2.0 equiv.) was added. The reaction was stirred at room temperature for 15 min. In a separate vial, Intermediate 30C (70 mg, 0.22 mmol, 1.0 equiv.) was dissolved in 0.5 mL of THF before adding 0.11 mL of iPrMgCl (2.0 M in THF, 0 .11 mmol, 1.0 equiv.). This solution was stirred for 5 minutes before applying it via syringe to the anilide reaction vial. The mixture was stirred at room temperature for 14 hours and diluted with EtOAc (30 mL), washed with 3 M HCl (2 x 15 mL) and brine, dried (MgSO4), filtered and concentrated. The crude residue was purified by silica gel chromatography (0% to 100% EtOAc in hexanes) to provide Example 30 (single diastereomer, relative stereochemistry undetermined and arbitrarily specified). 1H NMR (400 MHz; CD3OD): δ 8.89 (d, J = 5.5 Hz, 1H) 8.31 (d, J = 8.6 Hz, 1H) 8.14 (d, J = 8, 4 Hz, 1H) 7.94 (s, 1H) 7.77-7.85 (m, 2H) 7.50 (d, J = 8.8 Hz, 2H) 7.19-7.28 (m, 2H) 3,503.64 (m, 1H) 2.75 (s, 2H) 1.93-2.10 (m, 4H) 1.73-1.89 (m, 4H). m/z 395.2 (M+H)+. Examples 31 and 32 Example 31: N-(4-fluorophenyl)-2-(cis-4-(isoquinolin-4-yloxy)cyclohexyl)acetamide trifluoroacetate Example 32: N-(4-fluorophenyl)-2-(trans -4-(isoquinolin-4-yloxy)cyclohexyl)acetamide trifluoroacetate Intermediate 31A. Ethyl 2-(4-hydroxycyclohexyl)acetate:

[00304] A solution of ethyl (2-(4-oxocyclohexyl)acetate (See References in General Procedure A for preparation) (1.0 g, 5.4 mmol) in methanol (40 mL) at room temperature was NaBH4 (0.60 g, 16 mmol) was added.The resulting solution was stirred open to air for 13 hours, after which it was diluted with CH2Cl2 (60 mL), washed with 1 M HCl (3 x 30 mL), dried over anhydrous MgSO4, filtered, and concentrated under reduced pressure to provide Intermediate 31A as a 1:3 mixture of cis and trans isomers as a clear colorless oil (1.0 g, 99% yield).Intermediate 31B. N-(4-Fluorophenyl)-2-(4-hydroxycyclohexyl)acetamide:

[00305] Prepared using General Procedure G employing intermediate 31A (1.0 g, 5.4 mmol) and 4-fluoroaniline (1.01 mL, 11 mmol). Purified using silica gel chromatography (50% to 100% EtOAc in hexanes) to provide Intermediate 31B as a 1:2 mixture of diastereomers (white solid, 772 mg, 57%). Examples 31 and 32: N-(4-fluorophenyl)-2-(cis-4-(isoquinolin-4-yloxy)cyclohexyl)acetamide trifluoroacetate and N-(4-fluorophenyl)-2-(trans -4- (isoquinolin-4-yloxy)cyclohexyl)acetamide

[00306] To a vial containing Intermediate 31B (200 mg, 0.80 mmol, 1.0 equiv.) and 4-isoquinolinol (170 mg, 1.2 mmol, 1.5 equiv.) was added toluene followed by cyanomethylenetributylphosphorane ( 0.31 mL, 1.2 mmol, 1.5 equiv.) dropwise. The reaction was stirred at 60 °C for 16 h before concentrating. The crude residue was purified by silica gel chromatography (0% to 100% EtOAc in hexanes) to provide a mixture containing Examples 31 and 32, which was also purified by preparative reverse phase HPLC (PHENOMENEX® Gemini-NX, 10 μ, C18, 110A, 250 x 30 mm, 20 mL/min, eluting with 0% to 100% acetonitrile in water with 0.1% TFA gradient during the first 30 minutes of the 40 minute cycle) to provide o Example 31 (cis-diastereomer, first elution) as the TFA salt. 1H NMR (400 MHz; CDCl3): δ 8.87 (s, 1H) 8.22 (d, J = 8.0 Hz, 1H) 8.07 (s, 1H) 7.94 (d, J = 8 .0 Hz, 1H) 7.56-7.73 (m, 2H) 7.42-7.52 (m, 2H) 7.39 (br. s., 1H) 6.95-7.06 (m , 2H) 4.85 (br. s., 1H) 2.33 (d, J = 7.0 Hz, 2H) 2.22 (d, J = 13.1 Hz, 2H) 2.11 (s, 1H) 1.66-1.79 (m, 4H) 1.51-1.66 (m, 2H). m/z 379.2 (M+H)+.

[00307] Another elution from the previous preparative reversed phase HPLC provided Example 32 (trans-diastereomer, second elution) as the TFA salt. 1H NMR (400 MHz; CDCl3): δ 8.86 (s, 1H) 8.19-8.25 (m, 1H) 8.09 (s, 1H) 7.93 (d, J = 8.0 Hz , 1H) 7.69 (ddd, J = 8.3, 7.0, 1.3 Hz, 1H) 7.57-7.63 (m, 1H) 7.45-7.54 (m, 2H) 6.94-7.07 (m, 2H) 4.37-4.51 (m, 1H) 2.24-2.40 (m, 3H) 1.96-2.09 (m, 4H) 1, 57-1.72 (m, 2H) 1,171.31 (m, 2H). m/z 379.2 (M+H)+. Example 33 N-(4-Cyanophenyl)-2-( cis -4-(quinolin-4-yloxy)cyclohexyl)acetamide Example 33: N-(4-Cyanophenyl)-2-( cis -4-(quinolin-4-yloxy)cyclohexyl)acetamide

[00308] To the mixture of N-(4-cyanophenyl)-2-(4-hydroxycyclohexyl)acetamide (prepared using General Procedure G of Intermediate 31A and 4-cyanoaniline) (200 mg, 0.775 mmol, 1.0 equiv.), 4-quinolinol (168 mg, 1.16 mmol, 1.5 equiv.), and PPh3 (609 mg, 2.33 mmol 3.0 equiv.) cooled to 0 °C was added DEAD (0.182 mL, 1.16 mmol, 1.5 equiv.) dropwise. The ice bath was removed, and the reaction mixture was allowed to warm to room temperature and stir for 16 hours. The mixture was then diluted with EtOAc, filtered through a pad of CELITE®, and concentrated under reduced pressure. The crude residue was purified using silica gel chromatography (0% to 100% EtOAc in hexanes) to provide Example 33 (cis-diastereomer), which was also purified by preparative reversed-phase HPLC (PHENOMENEX® Gemini-NX, 10 μ, C18, 110A, 250 x 30 mm, 20 mL/min, eluting with 0% to 100% acetonitrile in water with a 0.1% TFA gradient during the first 30 minutes of the 40 minute cycle) to provide the Example 33 as the TFA salt. 1H NMR (400 MHz; CD3OD): δ 8.96 (d, J = 6.8 Hz, 1H) 8.48-8.56 (m, 1H) 8,068.17 (m, 2H) 7.90 (s , 1H) 7.76-7.83 (m, 2H) 7.62-7.69 (m, 2H) 7.52 (d, J = 6.8 Hz, 1H) 5.33 (br. s. , 1H) 2.65 (d, J = 0.6 Hz, 1H) 2.43 (d, J = 7.2 Hz, 2H) 2.25 (br. s., 2H) 2.05-2, 19 (m, 1H) 1.93 (br. s., 2H) 1.79 (d, J = 3.3 Hz, 2H) 1.64 (br. s., 2H). m/z 386.2 (M+H)+. Example 34 N-(4-Chlorophenyl)-2-( trans -1-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetamide (simple diastereomer, relative stereochemistry not determined and arbitrarily specified) Example 34: N-(4-Chlorophenyl)-2-(trans -1-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetamide (simple diastereomer, relative stereochemistry not determined and arbitrarily specified)

[00309] To a reaction vial containing 4-chloroaniline (33 mg, 0.25 mmol, 2.0 equiv.) was added iPrMgCl (2.0 M in THF, 0.13 mL, 0.25 mmol, 2, 0 equiv.). The reaction was stirred at room temperature for 15 min. In a separate vial, a single diastereomer obtained from Intermediate 30Ca (40 mg, 0.13 mmol, 1.0 equiv.) was dissolved in 0.5 mL of THF before adding 0.064 mL of iPrMgCl (2.0 M in THF, 0.13 mmol, 1.0 equiv.). This solution was stirred for 5 minutes before applying it with a syringe to the anilide reaction vial. The mixture was stirred at room temperature for 14 hours and diluted with EtOAc (30 mL), washed with 3 M HCl (2 x 15 mL) and brine, dried (MgSO4), filtered and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography (0% to 100% EtOAc in hexanes) to provide Example 34 (single diastereomer, relative stereochemistry undetermined and arbitrarily specified). 1H NMR (400 MHz; CD3OD): δ 9.09 (d, J = 5.9 Hz, 1H) 8.62 (d, J = 8.6 Hz, 1H) 8.20-8.29 (m, 1H) 8.15 (ddd, J = 8.4, 7.1, 1.1 Hz, 1H) 7.94-8.06 (m, 2H) 7.54-7.62 (m, 2H) 7 .23-7.33 (m, 2H) 3.74 (t, J = 12.1 Hz, 1H) 3.30 (dt, J = 3.3, 1.6 Hz, 1H) 2.61 (s , 2H) 2.09-2.26 (m, 2H) 1.98-2.09 (m, 2H) 1.81-1.96 (m, 4H). m/z 395.2 (M+H)+. Example 35 N-(4-Fluorophenyl)-2-( trans -1-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetamide (simple diastereomer, relative stereochemistry not determined and arbitrarily specified) Example 35: N-(4-Fluorophenyl)-2-(trans -1-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetamide (simple diastereomer, relative stereochemistry not determined and arbitrarily specified)

[00310] To a reaction vial containing 4-fluoroaniline (28 mg, 0.25 mmol, 2.0 equiv.) was added iPrMgCl (2.0 M in THF, 0.13 mL, 0.26 mmol, 2, 0 equiv.). The reaction was stirred at room temperature for 15 min. In a separate vial, a single diastereomer obtained from Intermediate 30Ca (40 mg, 0.13 mmol, 1.0 equiv.) was dissolved in 0.5 mL of THF before adding 0.064 mL of iPrMgCl (2.0 M in THF, 0.13 mmol, 1.0 equiv.). This solution was stirred for 5 minutes before applying it with a syringe to the anilide reaction bottle. The mixture was stirred at room temperature for 14 hours and diluted with EtOAc (30 mL), washed with 3 M HCl (2 x 15 mL) and brine, dried (MgSO4), filtered, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography (0% to 100% EtOAc in hexanes) to provide Example 35 (single diastereomer, relative stereochemistry undetermined and arbitrarily specified). 1H NMR (400 MHz; CDCl3): δ 8.81 (d, J = 4.7 Hz, 1H) 8.63-8.70 (m, 1H) 8.12 (dd, J = 8.4, 1 .0 Hz, 1H) 8.05 (d, J = 8.2 Hz, 1H) 7.63-7.73 (m, 1H) 7.42-7.60 (m, 3H) 7.34 (d , J = 4.7 Hz, 1H) 6.94-7.05 (m, 2H) 3.28 (t, J = 12.0 Hz, 1H) 2.58 (s, 2H) 1,972.16 (m , 4H) 1.84 (d, J = 10.9 Hz, 2H) 1.57-1.70 (m, 2H). m/z 379.2 (M+H)+. Example 36 N-(4-Chlorophenyl)-2-(trans-1-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetamide (simple diastereomer, relative stereochemistry not determined and arbitrarily specified) Example 36: N-(4-Chlorophenyl)-2-( trans -1-hydroxy-4-(quinolin-4-yl) cyclohexyl) acetamide (simple diastereomer, relative stereochemistry not determined and arbitrarily specified)

[00311] To a reaction vial containing 4-chloroaniline (51 mg, 0.40 mmol, 2.0 equiv.) was added iPrMgCl (2.0 M in THF, 0.20 mL, 0.40 mmol, 2, 0 equiv.). The reaction was stirred at room temperature for 15 min. In a separate vial, the cis-diastereomer obtained from Intermediate 30C (63 mg, 0.20 mmol, 1.0 equiv.) was dissolved in 0.5 mL of THF before adding 0.10 mL of iPrMgCl (2, 0 M in THF, 0.10 mmol, 1.0 equiv.). This solution was stirred for 5 minutes before applying it with a syringe to the anilide reaction bottle. The mixture was stirred at room temperature for 14 hours and diluted with EtOAc (30 mL), washed with 3 M HCl (2 x 15 mL) and brine, dried (MgSO4), filtered and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography (0% to 100% EtOAc in hexanes) to provide Example 36 (single diastereomer, relative stereochemistry undetermined and arbitrarily specified). 1H NMR (400 MHz; CDCl3): δ 9.73 (s, 1H) 8.75 (d, J = 4.5 Hz, 1H) 8.10 (dd, J = 8.4, 1.0 Hz, 1H) 8.03 (d, J = 7.8 Hz, 1H) 7.70 (ddd, J = 8.3, 7.0, 1.3 Hz, 1H) 7.50-7.62 (m, 3H) 7,217.29 (m, 2H) 7.14 (d, J = 4.7 Hz, 1H) 3.30-3.45 (m, 1H) 2.75 (s, 2H) 2.03-2 .12 (m, 2H) 1.93-2.02 (m, 2H) 1.82 (td, J = 13.3, 3.3 Hz, 2H) 1.47-1.63 (m, 2H) . m/z 395.2 (M+H)+. Example 37 N-(4-Fluorophenyl)-2-( trans -1-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetamide (simple diastereomer, relative stereochemistry not determined and arbitrarily specified) Example 37: N-(4-Fluorophenyl)-2-(trans -1-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetamide (simple diastereomer, relative stereochemistry not determined and arbitrarily specified)

[00312] To a reaction vial containing 4-fluoroaniline (44 mg, 0.40 mmol, 2.0 equiv.) was added iPrMgCl (2.0 M in THF, 0.20 mL, 0.40 mmol, 2, 0 equiv.). The reaction was stirred at room temperature for 15 min. In a separate vial, the cis-diastereomer obtained from Intermediate 30Cb (63 mg, 0.20 mmol, 1.0 equiv.) was dissolved in 0.5 mL of THF before adding 0.064 mL of iPrMgCl (2.0 M in THF, 0.10 mL, 0.20 mmol, 1.0 equiv.). This solution was stirred for 5 minutes before applying it with a syringe to the anilide reaction vial. The mixture was stirred at room temperature for 14 hours and diluted with EtOAc (30 mL), washed with 3 M HCl (2 x 15 mL) and brine, dried (MgSO4), filtered, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography (0% to 100% EtOAc in hexanes) to provide Example 37 (single diastereomer, relative stereochemistry undetermined and arbitrarily specified). 1H NMR (400 MHz; CDCl3): δ 9.60 (s, 1H) 8.76 (d, J = 4.7 Hz, 1H) 8.10 (dd, J = 8.4, 1.0 Hz, 1H) 8.04 (d, J = 8.0 Hz, 1H) 7.71 (ddd, J = 8.3, 6.9, 1.4 Hz, 1H) 7.52-7.65 (m, 2H) 7.15 (d, J = 4.7 Hz, 1H) 6.93-7.07 (m, 2H) 3.30-3.45 (m, 1H) 2.75 (s, 2H) 2 .07 (d, J = 13.5 Hz, 2H) 1.92-2.02 (m, 2H) 1.83 (td, J = 13.3, 3.4 Hz, 2H) 1.45-1 .65 (m, 2H). m/z 379.2 (M+H)+. Examples 38 and 39 (R)-N-(4-Cyanophenyl)-2-( cis -4-(quinolin-4-yloxy)cyclohexyl)propanamide (single enantiomer, absolute stereochemistry not determined and arbitrarily specified) (S)-N-(4-Cyanophenyl)-2-( cis -4-(quinolin-4-yloxy)cyclohexyl)propanamide (single enantiomer, absolute stereochemistry not determined and arbitrarily specified) Intermediate 38A: Ethyl 2-(4-((tert-butyldimethylsilyl)oxy)cyclohexyl)acetate

[00313] To a solution of ethyl 2-(4-hydroxycyclohexyl)acetate (17.4 g, 93.4 mmol, 1.0 equiv.) in DMF (100 mL) under argon was added imidazole (9.54 g , 140 mmol, 1.5 equiv.) and tert-butyldimethylsilyl chloride (15.5 g, 103 mmol, 1.1 equiv.). The reaction became cloudy and was stirred overnight. The reaction mixture was diluted with water (100 mL) and extracted with EtOAc (150 mL and 2 x 60 mL). The combined organic layers were dried (MgSO4) and concentrated to provide Intermediate 38A as a clear colorless oil. Intermediate 38B: Ethyl 2-(4-((tert-butyldimethylsilyl)oxy)cyclohexyl)propanoate

[00314] Intermediate 38A (5.0 g, 17 mmol, 1.0 equiv.) was diluted with Et2O and cooled to -78 °C. Sodium hexamethyldisilazide (2.0 M in THF, 9.2 mL, 18 mmol, 1.1 equiv.) was added via syringe, and the reaction turned light yellow and was stirred for 5 min. Methyl iodide (5.1 mL, 83 mmol, 5.0 equiv.) was added, and the reaction was allowed to warm to room temperature overnight. The reaction was diluted with EtOAc (100 mL) and brine (100 mL) and aq. Sat. (100ml). The layers were separated, and the aqueous layer was extracted with EtOAc (2 x 60 mL). The combined organic layers were dried (MgSO4), filtered, concentrated under reduced pressure, and purified by silica gel chromatography (0% to 10% EtOAc in hexanes) to provide Intermediate 38B as a faint yellow oil (4.56 g , 86%). Intermediate 38C: Ethyl 2-(4-hydroxycyclohexyl)propanoate

[00315] Intermediate 38B (4.54 g, 14.5 mmol, 1.0 equiv.) was diluted in THF (48 mL), and the solution was treated with tetrabutylammonium fluoride solution (1.0 M in THF, 22 mL, 21.7 mmol, 1.5 equiv.). The reaction solution was allowed to stir overnight. The reaction solution was concentrated and then diluted with EtOAc (200 mL), washed with water and brine, dried (MgSO4), filtered, and concentrated under reduced pressure. The crude residue was purified by silica gel chromatography (0% to 30% EtOAc in hexanes) to provide Intermediate 38C as a trans to cis ratio of 7:3 (2.78 g, 96% production). Intermediate 38D: ethyl 2-(cis-4-(quinolin-4-yloxy)cyclohexyl)propanoate

[00316] To a solution of Intermediate 38C (1.07 g, 5.35 mmol, 1.0 equiv.), 4-quinolinol (0.931 g, 6.42 mmol, 1.2 equiv.), and PPh3 (4 .22 g, 16.1 mmol, 3.0 equiv.) in THF (18 mL, 0.3 M) at 0 °C was added DEAD (1.27 mL, 8.03 mmol, 1.5 equiv.) . The reaction was allowed to warm to room temperature overnight. The reaction was concentrated and then diluted with EtOAc (100 mL). The solution was washed with 1 M NaOH and brine, dried (MgSO4), filtered, concentrated under reduced pressure, and purified by silica gel chromatography (0% to 100% EtOAc in hexanes) to provide Intermediate 38D. Examples 38 and 39: (R)-N-(4-Cyanophenyl)-2-( cis -4-(quinolin-4-yloxy)cyclohexyl)propanamide and (S)-N-(4-Cyanophenyl)-2-( cis-4-(quinolin-4-yloxy)cyclohexyl)propanamide (both as single enantiomers, absolute stereochemistry not determined and arbitrarily specified)

[00317] To a solution of 4-chloroaniline (340 mg, 2.9 mmol, 2.0 equiv.) in THF (2.5 mL) was added iPrMgCl (2.0 M in THF, 1.45 mL, 2 .9 mmol, 2.0 equiv.). The solution turned brown/orange and was stirred for 15 min. A solution of intermediate 38D (502 mg, 1.45 mmol, 1.0 equiv.) in THF (0.5 mL) was added, and the reaction was allowed to stir overnight. The reaction mixture was diluted with EtOAc (30 mL) and washed with 3 M HCl (2 x 15 mL) and brine. The combined organics were dried (MgSO4), filtered, concentrated under reduced pressure, and purified by silica gel chromatography (0% to 100% EtOAc) to provide Examples 38 and 39 as a mixture of enantiomers. Chiral semipreparative normal phase HPLC (Diacel CHIRALPAK® ID, 5 μ, 250 x 20 mm, 15 mL/min, eluting with 95% 1:1 hexane:CH2Cl2 and 5% acetonitrile with 0.4% isocratic diethylamine for 40 min) to provide Example 38 as the first eluting isomer. 1H NMR (400 MHz; CDCl3): δ 8.71 (d, J = 5.3 Hz, 1H) 8.19 (dd, J = 8.4, 1.4 Hz, 1H) 8.02 (d, J = 8.4 Hz, 1H) 7.78 (s, 1H) 7.64-7.73 (m, 2H) 7.54-7.61 (m, 2H) 7.44 (ddd, J = 8 .2, 7.0, 1.2 Hz, 1H) 6.70 (d, J = 5.5 Hz, 1H) 4.86 (br. s., 1H) 2.14-2.32 (m, 2H) 1.47-1.86 (m, 7H), 1.27 (d, J = 7.0 Hz, 2H). m/z 400.2 (M+H)+.

[00318] Another elution from the previous semipreparative chiral HPLC column provided Example 39 as the second elution isomer. 1H NMR (400 MHz; CDCl3): δ 8.71 (d, J = 5.3 Hz, 1H) 8.19 (dd, J = 8.4, 1.4 Hz, 1H) 8.02 (d, J = 8.4 Hz, 1H) 7.78 (s, 1H) 7.64-7.73 (m, 2H) 7.54-7.61 (m, 2H) 7.44 (ddd, J = 8 .2, 7.0, 1.2 Hz, 1H) 6.70 (d, J = 5.5 Hz, 1H) 4.86 (br. s., 1H) 2.14-2.32 (m, 2H) 1.47-1.86 (m, 7H), 1.27 (d, J = 7.0 Hz, 2H). m/z 400.2 (M+H)+. Examples 40 and 41 (R)-N-(4-Fluorophenyl)-2-( cis -4-(quinolin-4-yloxy)cyclohexyl)propanamide (single enantiomer, absolute stereochemistry not determined and arbitrarily specified) (S)-N-(4-Fluorophenyl)-2-( cis -4-(quinolin-4-yloxy)cyclohexyl)propanamide (single enantiomer, absolute stereochemistry not determined and arbitrarily specified)

[00319] Examples 40 and 41: (R)-N-(4-Fluorophenyl)-2-(cis-4-(quinolin-4-yloxy)cyclohexyl) propanamide and (S)-N-(4-Fluorophenyl)- 2-(cis-4-(quinolin-4-yloxy)cyclohexyl) propanamide (both obtained as a single enantiomer, absolute stereochemistry not determined and arbitrarily specified)

[00320] To a solution of 4-fluoroaniline (0.275 mL, 2.9 mmol, 2.0 equiv.) in THF (2.5 mL) was added iPrMgCl (2.0 M in THF, 1.45 mL, 2 .9 mmol, 2.0 equiv.). The solution turned brown/orange and was stirred for 15 min. A solution of intermediate 38D (502 mg, 1.45 mmol, 1.0 equiv.) in THF (0.5 mL) was added, and the reaction was allowed to stir overnight. The reaction mixture was diluted with EtOAc (30 mL) and washed with 3 M HCl (2 x 15 mL) and brine. The solution was dried (MgSO4), concentrated, and purified by silica gel chromatography (0% to 100% EtOAc) to provide Examples 40 and 41 as a mixture of enantiomers. Chiral semipreparative normal phase HPLC (Diacel CHIRALPAK® ID, 5 μ, 250 x 20 mm, 15 mL/min, eluting with 95% 1:1 hexane:CH2Cl2 and 5% acetonitrile with 0.4% isocratic diethylamine for 40 min) to provide Example 40 as the first eluting isomer. 1H NMR (400 MHz; CDCl3): δ 8.72 (d, J = 5.3 Hz, 1H) 8.22 (dd, J = 8.3, 1.1 Hz, 1H) 8.03 (d, J = 8.6 Hz, 1H) 7.69 (ddd, J = 8.4, 6.9, 1.5 Hz, 1H) 7.42-7.53 (m, 2H) 7.21 (s, 1H) 6.94-7.05 (m, 2H) 6.71 (d, J = 5.5 Hz, 1H) 4.87 (br. s., 1H) 2,062.31 (m, 4H) 1, 47-1.88 (m, 5H) 1.22-1.29 (m, 4H). m/z 393.2 (M+H)+.

[00321] Another elution from the previous semipreparative chiral HPLC column provided Example 41 as the second elution isomer. 1H NMR (400 MHz; CDCl3): δ 8.72 (d, J = 5.3 Hz, 1H) 8.22 (dd, J = 8.3, 1.1 Hz, 1H) 8.03 (d, J = 8.6 Hz, 1H) 7.69 (ddd, J = 8.4, 6.9, 1.5 Hz, 1H) 7.42-7.53 (m, 2H) 7.21 (s, 1H) 6.94-7.05 (m, 2H) 6.71 (d, J = 5.5 Hz, 1H) 4.87 (br. s., 1H) 2.06-2.31 (m, 4H) 1.47-1.88 (m, 5H) 1.22-1.29 (m, 4H). m/z 393.2 (M+H)+. Example 42 (±)-N-(4-chlorophenyl)-2-( trans -4-(quinolin-4-yl)cyclohexyl)pent-4-enamide Intermediate 42A: ethyl-2-4-(quinolin-4-yl)cyclohexyl)pent-4-enoate

[00322] To a solution of ethyl 2-(4-(quinolin-4-yl)cyclohexyl)acetate (which can be prepared using General Procedures A and B) in THF (0.2 M) at 0° NaHMDS solution (2 equivalents) was added. The resulting yellow solution was stirred at 0°C for 5 minutes and allyl bromide (2 equivalents) was added. The reaction mixture was warmed to room temperature and stirred for 1 hour, under which AcOH was added along with Et2O. The reaction mixture was filtered through a silica plug eluting with additional Et2O. The filtrate was concentrated and purified using silica gel chromatography to provide Intermediate 42A as a mixture of cis/trans diastereomers. Example 42: N-(4-chlorophenyl)-2-(trans-4-(quinolin-4-yl)cyclohexyl)pent-4-enamide

[00323] Intermediate 42A was subjected to General Procedure G using 4-chloroaniline and Example 42 was isolated as a white solid. 1H NMR (400 MHz, CDCl3): δ 9.53 (s, 1H), 8.58 (d, J = 4.6 Hz, 1H), 8.10 (dd, J = 11.5, 8.5 Hz, 2H), 7.70 (t, J = 7.2 Hz, 1H), 7.60 (t, J = 7.2 Hz, 1H), 7.54 (d, J = 8.8 Hz, 2H), 7.25 (ddd, J = 8.9, 5.8, 2.3 Hz, 1H), 7.20 - 7.15 (m, 2H), 6.00 - 5.87 (m, 1H), 5.21 - 5.09 (m, 2H), 3.46 (s, 1H), 2.71 (td, J = 10.1, 4.9 Hz, 1H), 2.45 (q , J = 9.7 Hz, 2H), 2.28 - 1.62 (m, 9H). Example 43 2-(4-(1H-pyrazolo[3,4-b]pyridin-4-yl)cyclohexyl)-N-(4-chlorophenyl)acetamide (mixture of cis/trans diastereomers) Example 43: 2-(4-(1H-pyrazolo[3,4-b]pyridin-4-yl)cyclohexyl)-N-(4-chlorophenyl)acetamide (mixture of cis/trans diastereomers)

[00324] Prepared using General Procedures K, B, and G. In General Procedure K, 4-bromo-1H-pyrazolo[3,4-b]pyridine was used as the coupling partner and dimethoxyethane was used as the solvent. In General Procedure G, 4-chloroaniline was used. Example 43 was isolated as a white solid and ~1:1 mixture of diastereomers. 1H NMR (400 MHz, CDCl3): δ 8.52 - 8.45 (m, 1H), 8.20 - 8.11 (m, 1H), 7.52 - 7.46 (m, 2H), 7 .34 - 7.27 (m, 2H), 7.08 - 6.96 (m, 1H), 3.20 - 2.90 (m, 1H), 2.50 (s, 1H), 2.33 (d, J = 6.7 Hz, 1H), 2.14 - 1.64 (m, 8H), 1.39 - 1.25 (m, 2H). LC/MS, m/z 369 (M+H)+. Examples 44 and 45 N-(4-fluorophenyl)-2-( cis -4-(quinazolin-4-yl)cyclohexyl)acetamide N-(4-Fluorophenyl)-2-(4-(quinazlin-4-yl)cyclohexyl)acetamide (mixture of diastereomers) Examples 44 and 45: N-(4-fluorophenyl)-2-( cis -4-(quinazolin-4-yl)cyclohexyl)acetamide and N-(4-fluorophenyl)-2-(4-(quinazlin-4-yl )cyclohexyl)acetamide (mixture of diastereomers)

[00325] Prepared using General Procedures K, B, and G. 4-Chloroquinazoline and Pd(PPh3)4 were used in Procedure K. 4-Fluoroaniline was used in Procedure G. Purified using silica gel chromatography (50% EtOAc in hexanes) to provide Example 44 (cis diastereomer) as the first eluting isomer. 1H NMR (400 MHz, CD3OD) δ 9.15 (s, 1H), 8.37 (d, J = 8.5 Hz, 1H), 8.08 - 7.93 (m, 2H), 7.75 (ddd, J = 8.4, 6.0, 2.2 Hz, 1H), 7.56 (ddd, J = 7.0, 5.3, 2.8 Hz, 2H), 7.08 - 6 .97 (m, 2H), 3.82 (t, J = 10.0 Hz, 1H), 2.56 (d, J = 7.9 Hz, 2H), 2.43 (s, 1H), 2 .20 - 2.02 (m, 2H), 2.00 (s, 1H), 2.00 - 1.74 (m, 6H) ppm. m/z 364.2 (M+H)+.

[00326] Further elution from the column provided Example 45 as a 1:1 mixture of cis:trans isomers. m/z 364.2 (M+H)+. Examples 46 and 47 Example 46: N-(4-chlorophenyl)-2-( cis -4-(quinazolin-4-yl)cyclohexyl)acetamide Example 47: N-(4-chlorophenyl)-2-(trans -4-(quinazlin-4-yl)cyclohexyl)acetamide Examples 46 and 47: N-(4-chlorophenyl)-2-( cis -4-(quinazolin-4-yl)cyclohexyl)acetamide and N-(4-chlorophenyl)-2-( trans -4-(quinazlin-4 - il)cyclohexyl)acetamide

[00327] Prepared using General Procedures K, B, and G. 4-Chloroquinazoline and Pd(PPh3)4 were used in Procedure K. 4-Chloroaniline was used in Procedure G. Purified using silica gel chromatography (50% EtOAc in hexanes) to provide Example 46 (cis-diastereomer) as the first eluting isomer. 1H NMR (400 MHz, CDCl3) δ 9.28 (s, 1H), 8.19 (d, J = 8.2 Hz, 1H), 8.06 (d, J = 8.3 Hz, 1H), 7.90 (ddd, J = 8.4, 6.9, 1.3 Hz, 1H), 7.67 (ddd, J = 8.3, 6.9, 1.2 Hz, 2H), 7, 53 (d, J = 8.8 Hz, 2H), 7.32 - 7.24 (m,2H), 3.69 (dd, J = 12.0, 8.6 Hz, 1H), 2.57 (s, 3H), 2.05 (dt, J = 19.0, 8.4 Hz, 2H), 1.86 - 1.72 (m, J = 30.7, 27.2 Hz, 6H) ppm . m/z 380 (M+H)+.

[00328] Another elution from the column provided Example 47 (trans diastereomer) as the second elution isomer. 1H NMR (400 MHz, CDCl3) δ 9.25 (s, 1H), 8.17 (d, J = 8.6 Hz, 1H), 8.05 (d, J = 8.3 Hz, 1H), 7.89 (t, J = 7.7 Hz, 1H), 7.65 (t, J = 7.7 Hz, 1H), 7.51 (d, J = 8.7 Hz, 2H), 7, 35 - 7.15 (m, 3H), 3.54 (t, J = 11.5 Hz, 1H), 2.36 (d, J = 6.5 Hz, 2H), 2.15 - 1.81 (m, 5H), 1.48 - 1.30 (m, 2H), 0.87 (d, J = 11.4 Hz, 2H) ppm. m/z 380 (M+H)+. Example 48 N-(4-chlorophenyl)-2-(4-(1,5-naphtyridin-4-yl)cyclohexyl)acetamide (mixture of diastereomers) Example 48: N-(4-chlorophenyl)-2-(4-(1,5-naphthyridin-4-yl)cyclohexyl)acetamide (mixture of diastereomers)

[00329] Prepared using General Procedures K, B, and G. 4-Chloro-1,5-naphthyridine was used in Procedure K and 4-chloroaniline was used in Procedure G. Purified using silica gel chromatography (25-75% of EtOAc in hexanes) to provide a residue. The residue was also purified using silica gel chromatography (25-75% EtOAc in toluene) to provide the desired product as a 2:1 mixture of diastereomers. m/z 380.2 (M+H)+. Example 49 N-(4-chlorophenyl)-2-( cis -4-(quinolin-3-yl)cyclohexyl)propanamide Example 49: N-(4-chlorophenyl)-2-( cis -4-(quinolin-3-yl)cyclohexyl)propanamide

[00330] Prepared with General Procedure G using ethyl 2-(4-(quinolin-3-yl)cyclohexyl)propanoate and 4-chloroaniline. Purified using silica gel chromatography (0% to 10% 2-propanol in hexanes) to provide Example 49 as a white solid and the first eluting isomer. 1H NMR (400 MHz, CDCl3): δ 8.74 (d, J=2.2Hz. 1H), 8.64 (s, 1H), 8.05-8.11 (m, 1H), 7.68 -7.75 (m, 2H), 7.55-7.64 (m, 3H), 7.23-7.28 (m, 3H), 2.74-2.84 (m, 1H), 2 .44-2.54 (m, 1H), 1.99-2.09 (m, 1H), 1.36-1.80 (m, 8H), 1.26 (d, J=6.9Hz, 3H) ppm. m/z 393.3 (M+H)+. Example 50 N-(4-fluorophenyl)-2-( trans -4-(quinolin-3-yl)cyclohexyl)propanamide Example 50: N-(4-fluorophenyl)-2-(trans -4-(quinolin-3-yl)cyclohexyl)propanamide

[00331] Prepared with General Procedure G using ethyl 2-(4-(quinolin-3-yl)cyclohexyl)propanoate and 4-fluoroaniline. Purified using silica gel chromatography (0% to 10% 2-propanol in hexanes) to provide Example 50 as a white solid and the second eluting isomer. 1H NMR (400 MHz, CDCl3): δ 8.80 (d, J=2.2Hz, 1H), 8.04-8.09 (m, 1H), 7.89-7.91 (m, 1H) , 7.75-7.80 (m, 1H), 7.52-7.69 (m, 1H), 7.48-7.56 (m, 3H), 7.27 (s, 1H), 6 .98-7.06 (2H), 2.69 (tt, 1H, J=3.1Hz, J=12.6Hz), 1.95-2.20 (m, 5H), 1.54-1, 90 (m, 5H), 1.28 (d, J=6.9Hz, 3H) ppm. m/z 377.3 (M+H)+. Examples 51 and 52 N-(4-Chlorophenyl)-2-((trans)-4-(isoquinolin-1-yl)cyclohexyl)acetamide N-(4-Chlorophenyl)-2-((cis)-4-(isoquinolin-1-yl)cyclohexyl)acetamide Examples 51 and 52: N-(4-Chlorophenyl)-2-((trans)-4-(isoquinolin-1-yl)cyclohexyl)acetamide and N-(4-Chlorophenyl)-2-((cis)-4- (isoquinolin-1-yl)cyclohexyl)acetamide

[00332] Prepared using General Procedures K, B, and G. For General Procedure K, 1-chloroisoquinoline was employed. For General Procedure B, ethanol was used as the solvent, and the reaction was carried out at 50 °C. For General Procedure G, 4-chloroaniline was employed, and trimethylaluminum was used in place of iPrMgCl. The desired compound was purified using preparative HPLC (Varian ProStar using Hamilton C18 PRP-1 column (15 x 250 mm) with a flow rate of 20 mL/min, Mobile phase A: 0.5% formic acid in water; Mobile phase B: 0.5% formic acid in acetonitrile; 0% to 100% gradient elution B over 30 minutes.) to provide examples 51 and 52 as a mixture of diastereomers. The residue was also purified by preparative TLC (33% EtOAc in hexanes) to provide Example 51 (trans diastereomer, higher retention factor). 1H NMR (400 MHz; CDCl3): δ 8.49-8.43 (m, 1H), 8.24-8.20 (m, 1H), 7.85-7.82 (m, 2H), 7 .65-7.50 (m, 4H), 7.28-7.25 (m, 2H), 3.70-3.62 (m, 1H), 2.62-2.56 (m, 2H) , 2.41-2.36 (m, 2H), 2.09-2.02 (m, 2H), 1.86-1.82 (m, 3H), 1.28-1.24 (m, 2H) ppm. m/z 379.2 (M+H)+.

[00333] Preparative TLC also provided Example 52 (cis diastereomer, lower retention factor). 1H NMR (400 MHz; CDCl3): δ 8.50-8.46 (m, 1H), 8.24-8.20 (m, 1H), 7.86-7.82 (m, 1H), 7 .72-7.50 (m, 4H), 7.30-7.26 (m, 2H), 3.59-3.54 (m, 1H), 2.38-2.33 (m, 2H) , 2.20-1.95 (m, 6H), 1.40-1.22 (m, 3H) ppm. m/z 379.1 (M+H)+. Examples 53 and 54 N-(4-Chlorophenyl)-2-((cis)-4-(isoquinolin-8-yl)cyclohexyl)acetamide N-(4-Chlorophenyl)-2-((trans)-4-(isoquinolin-8-yl)cyclohexyl)acetamide Examples 53 and 54: N-(4-Chlorophenyl)-2-(( cis )-4-(isoquinolin-8-yl)cyclohexyl)acetamide and N-(4-Chlorophenyl)-2-(( trans )-4- (isoquinolin-8-yl)cyclohexyl)acetamide

[00334] Prepared using General Procedures K, B, and G. For General Procedure K, 8-bromoisoquinoline was employed. For General Procedure B, ethanol was used as the solvent and the reaction was carried out at 50 °C. For General Procedure G, 4-chloroaniline was employed and trimethylaluminum was used in place of iPrMgCl. The residue was purified using preparative HPLC (Varian ProStar using a Hamilton C18 PRP-1 column (15 x 250 mm) with a flow rate of 20 mL/min, Mobile phase A: 0.5% formic acid in water ; Mobile phase B: 0.5% formic acid in acetonitrile; 0% to 100% gradient elution B over 30 minutes) to provide Example 53 (cis diastereomer) as the first eluting isomer. 1H NMR (400 MHz; CDCl3): δ 9.25 (s, 1H), 8.56-8.53 (m, 1H), 7.85-7.82 (m, 2H), 7.62-7 .47 (m, 4H), 7.31-7.26 (m, 2H), 3.38-3.27 (m, 1H), 2.572.53 (m, 3H), 2.04-1.25 (m, 8H) ppm. m/z 379 (M+H)+.

[00335] Another elution provided Example 54 (trans diastereomer) as the second elution isomer. 1H NMR (400 MHz; CDCl3): δ 9.24 (s, 1H), 8.56-8.52 (m, 1H), 7.86-7.81 (m, 2H), 7.69-7 .49 (m, 4H), 7.32-7.26 (m, 2H), 3.26-3.18 (m, 1H), 2.35-2.23 (m, 3H), 1.67 -1.59 (m, 4H), 1.40-1.27 (m, 4H) ppm. m/z 379 (M+H)+. Example 55 N-(4-Chlorophenyl)-2-((trans)-4-(isoquinolin-5-yl)cyclohexyl)acetamide (simple diastereomer, relative stereochemistry not determined and arbitrarily specified) Example 55: N-(4-Chlorophenyl)-2-((trans)-4-(isoquinolin-5-yl)cyclohexyl)acetamide

[00336] Prepared using General Procedures K, B, and G. For General Procedure K, 5-bromoisoquinoline was employed. For General Procedure B, ethanol was used as the solvent, and the reaction was carried out at 50 °C. For General Procedure G, 4-chloroaniline was employed, and trimethylaluminum was used in place of iPrMgCl. The residue was purified using preparative HPLC (Varian ProStar using a Hamilton C18 PRP-1 column (15 x 250 mm) with a flow rate of 20 mL/min. Mobile phase A: 0.5% formic acid in water; Phase mobile B: 0.5% formic acid in acetonitrile; 0% to 100% gradient elution B over 30 minutes) to provide Example 55 (simple diastereomer, relative stereochemistry undetermined and arbitrarily specified). 1H NMR (400 MHz; CDCl3): δ 9.24 (s, 1H), 8.55-8.52 (m, 1H), 7.85-7.81 (m, 2H), 7.60-7 .48 (m, 4H), 7.32-7.26 (m, 2H), 3.27-3.20 (m, 1H), 2.38-2.32 (m, 1H), 2.09 -2.02 (m, 3H), 1.70-1.57 (m, 3H), 1.41-1.24 (m, 4H). m/z 379 (M+H)+. Example 56 N-(4-fluorophenyl)-2-( cis -4-(quinolin-3-yl)cyclohexyl)propanamide Example 56: N-(4-fluorophenyl)-2-( cis -4-(quinolin-3-yl)cyclohexyl)propanamide

[00337] Prepared with General Procedure G using ethyl 2-(4-(quinolin-3-yl)cyclohexyl)propanoate and 4-fluoroaniline. Purified using silica gel chromatography (0% to 10% 2-propanol in hexanes) to provide Example 56 (cis diastereomer) as a white solid and the first eluting isomer. 1H NMR (400 MHz, CDCl3): δ 8.76-8.79 (m, 1H), 8.19 (bs, 1H), 8.06-8.11 (m, 1H), 7.65-7 .77 (m, 3H), 7.53-7.62 (m, 3H), 6.96-7.04 (m, 2H), 2.79-2.89 (m, 1H), 2.43 -2.53 (m, 1H), 1.99-2.08 (m, 1H), 1.50-1.85 (m, 8H), 1.27 (d, J=6.8Hz, 3H) ppm. m/z 377.3 (M+H)+. Example 57 (±)-N-(4-Cyanophenyl)-2-((cis)-4-(quinolin-4-yl)cyclohexyl)pent-4-enamide Intermediate 57A: Ethyl 2-(cis-4-(quinolin-4-yl)cyclohexyl)pent-4-enoate

[00338] To a solution of ethyl 2-(4-(quinolin-4-yl)cyclohexyl)acetate (which can be prepared using General Procedures A and B) (740 mg, 2.5 mmol) in THF (5 mL) at -78 °C NaHMDS (2 M in THF, 3.0 mL, 6.0 mmol) was added. The resulting reaction mixture was stirred at -78°C for 5 min, and allyl bromide (333 mg, 2.75 mmol) was added. The reaction was warmed to 0 °C and stirred for 10 min. The reaction was diluted with Et2O and filtered through a 2x2 cm pad of silica gel, washing with another 50 mL of Et2O. The filtrate was concentrated and used directly in the next step. Example 57: (±)-N-(4-Cyanophenyl)-2-((cis)-4-(quinolin-4-yl)cyclohexyl)pent-4-enamide

[00339] Intermediate 57A was hydrolyzed to the carboxylic acid using General Procedure E. The acid product was coupled with 4-cyanoaniline using General Procedure O. Example 57 was obtained as a white solid after column chromatography (10% at 30 % 2-propanol in hexanes). 1H NMR (400 MHz; CDCl3): δ 9.16 (s, 1H), 8.67 (d, J = 4.6 Hz, 1H), 8.13-8.08 (m, 2H), 7, 72 (dtd, J = 8.7, 3.4, 1.7 Hz, 3H), 7.61 (ddd, J = 8.4, 6.9, 1.4 Hz, 1H), 7.55- 7.51 (m, 2H), 7.14 (d, J = 4.6 Hz, 1H), 5.95-5.84 (m, 1H), 5.18 (dt, J = 16.3, 1.1Hz, 1H), 5.13-5.10 (m, 1H), 3.50-3.49 (m, 1H), 2.69-2.68 (m, 1H), 2.48 -2.41 (m, 2H), 2.23 (dt, J = 10.4, 0.3 Hz, 1H), 2.01-1.65 (m, 8H). Example 58 (R )-2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)-N-phenylpropanamide Intermediate 58A: (R)-2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)propanoic acid

[00340] Intermediate 58A can be prepared from General Procedures K, B, E, L, M, and N. General Procedure L employed 2-(4-(6-fluoroquinolin-4-yl)-cyclohexyl)acetic acid (mixture of diastereomers) and (R)-2-phenyl-oxazolidinone. General Procedure M employed the cis product and iodomethane. The auxiliary was removed after General Procedure No. LC-MS Anal: m/z [M+H]+ 302.2. 1H-NMR (400 MHz; DMSO, d6): δ 12.10 (s, 1H), 8.70 (d, J = 4.5 Hz, 1H), 8.07 (dd, J = 9.2, 5.9 Hz, 1H), 7.97-7.94 (m, 1H), 7.67-7.62 (m, 1H), 7.49 (d, J = 4.5 Hz, 1H), 3,413.36 (m, 1H), 2.73-2.65 (m, 1H), 1.83-1.61 (m, 9H), 1.08 (d, J = 6.8 Hz, 3H) . Example 58: (R )-2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)-N-phenylpropanamide

[00341] Intermediate 58A, (R)-2-((1s,4S)-4-(6-fluoroquinolin-4-yl)cyclohexyl)propanoic acid (10 mg, 0.033 mmol) was added to a vial and seized in DMF (350 μl). Aniline (4.64 mg, 0.050 mmol) and HATU (18.93 mg, 0.050 mmol) were added, followed by DIPEA (17.39 μl, 0.100 mmol). The reaction was stirred at room temperature for 3 hours. The reaction mixture was diluted with DMF to reach a total volume of 2 mL and filtered. The crude sample was purified by preparative HPLC/MS to provide Example 58 (9.7 mg, 0.026 mmol, 78%). LC-MS Anal. Calculated for C24H25FN2O 376.20, found [M+H] 377.0 Tr = 1.969 minutes (Method B). 1H NMR (500 MHz, DMSO-d6) δ: 9.96 (s, 1H), 8.82 (d, J=4.5 Hz, 1H), 8.05 (dd, J=9.2, 5 .8 Hz, 1H), 7.93 (dd, J=10.9, 2.5 Hz, 1H), 7.62 (td, J=8.7, 2.6 Hz, 1H), 7.57 (d, J=7.9 Hz, 2H), 7.53 (d, J=4.5 Hz, 1H), 7.25 (t, J=7.8 Hz, 2H), 6.99 (t , J=7.4 Hz, 1H), 2.83 (dd, J=10.8, 6.7 Hz, 1H), 1.64-1.97 (m, 7H), 1.51-1, 64 (m, 3H), 1.08 (d, J=6.6 Hz, 3H) Examples 59 to 81

[00342] Examples 59-81 were prepared from Intermediate 58A following the procedure for example 58 using the corresponding anilines or aryl amines. Table 1 Example 82 (R)-N-(3-Cyano-4-(trifluoromethyl)phenyl)-2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)propanamide

[00343] Intermediate 58A (15 mg, 0.050 mmol) was taken up in tetrahydrofuran (249 μl) and triethylamine (13.88 μl, 0.100 mmol) was added. The solution was cooled to 0 °C. Isobutylcarbonochloridate (10.20 mg, 0.075 mmol) was added and the reaction stirred at 0 °C for 10 minutes before adding 5-amino-2-(trifluoromethyl)benzonitrile (12.97 mg, 0.070 mmol). After addition of aniline, the reaction was allowed to warm to room temperature and stir at room temperature overnight. After 16 hours, 5-amino-2-(trifluoromethyl)benzonitrile (12.97 mg, 0.070 mmol) was added and the reaction was heated to 50 °C for 24 hours. The reaction was then concentrated in vacuo, diluted with DMF (~2 mL), filtered, and purified by preparative HPLC to provide Example 82 (5.8 mg, 0.012 mmol, 25%). LC-MS Anal. Calculated for C26H23F4N3O 469.18, found [M+H] 470.1 Tr = 2.219 minutes (Method B). 1H NMR (500 MHz, DMSO-d6) δ: 10.69 (br. s., 1H), 8.79 (br. s., 1H), 8.38 (br. s., 1H), 8, 05 (d, J=8.2 Hz, 2H), 7.95 (d, J=8.0 Hz, 2H), 7.60-7.71 (m, 1H), 7.44 (br. s ., 1H), 2.39 (br. s., 1H), 1.83-2.00 (m, 3H), 1.60-1.83 (m, 3H), 1.27-1.60 (m, 4H), 1.15 (d, J=6.2 Hz, 3H) Example 83 (±)-N-(4-Chlorophenyl)-2-(( trans )-4-((7-(trifluoromethyl )quinolin-4-yl)oxy)cyclohexyl)butanamide Intermediate 83A: Ethyl 2-(1,4-dioxaspiro[4,5]decan-8-ylidene)acetate

[00344] Triethyl phosphonoacetate (21.79 ml, 109 mmol) was added to a suspension of sodium hydride (3.84 g, 96 mmol) in THF (64.0 ml) at 0 °C. The reaction was stirred at room temperature for 30 minutes. After 30 minutes, the reaction was cooled again to 0 °C and a solution of 1,4-dioxaspiro[4,5]decan-8-one (10 g, 64.0 mmol) in 5 mL of THF was added. The reaction was then stirred at room temperature for 30 minutes before quenching with water. The mixture was extracted with DCM three times. Combined organic extracts were dried with sodium sulfate, filtered, and concentrated in vacuo. The crude residue was purified by silica gel chromatography to provide Intermediate 83A (13.88 g, 61.3 mmol, 96% yield). TLC: product spots as purple dots in anisaldehyde (Rf = 0.75 in 1:1 Hex/EtOAc). 1H NMR (400 MHz, chloroform-d) δ: 5.65 (s, 1H), 4.13 (q, J=7.2 Hz, 2H), 3.92-3.99 (m, 4H ), 2.94-3.02 (m, 2H), 2.31-2.40 (m, 2H), 1.71-1.79 (m, 4H), 1.26 (t, J=7 .2 Hz, 3H) Intermediate 83B: ethyl 2-(1,4-dioxaspiro[4,5]decan-8-yl)acetate

[00345] Intermediate 83A (13.88 g, 61.3 mmol) was taken up in EtOAc (61.3 ml) and was added to a Parr hydrogenation flask containing 10% palladium on wet carbon (1.306 g, 12.27 mmol) (54% w/w water) under a nitrogen atmosphere. The reaction flask was purged and recharged with nitrogen three times, and then purged and recharged three times with hydrogen. After charging the flask with hydrogen to 3.51 kg/cm2 (50 psi), the flask was placed on a Parr shaker and shaken. After 4 hours, the reaction was filtered over pressed CELITE® and concentrated in vacuo to provide Intermediate 83B (13.78 g, 60.4 mmol, 98% yield). LC-MS Anal. Calculated for C12H20O4 228.14, found [M+H] 299.1 Tr = 0.83 minutes (Method A). 1H NMR (400 MHz, chloroform-d) δ: 4.11 (q, J=7.2 Hz, 2H), 3.88-3.95 (m, 4H), 2.21 (d, J =7.0 Hz, 2H), 1.83 (dqd, J=11.0, 7.5, 3.5 Hz, 1H), 1.68-1.78 (m, 4H), 1,501.61 ( m, 2H), 1.27-1.35 (m, 2H), 1.24 (t, J=7.2 Hz, 3H). Intermediate 83C: Ethyl 2-(1,4-dioxaspiro[4,5]decan-8-yl)butanoate

[00346] Diisopropylamine (2.347 ml, 16.63 mmol) taken up in dry THF (15.99 ml) (under N2 atmosphere) and cooled to -78 °C. n - BuLi (6.14 ml, 15.35 mmol) (2.5 M in hexanes) was added over ~5 minutes at -78 °C. After stirring for 45 minutes, the reaction was warmed to room temperature for 10 minutes and returned to -78 °C. Then, 1,3-dimethyltetrahydropyrimidin-2(1H)-one (1.541 ml, 12.79 mmol) was added, followed by a solution of intermediate 83B (2.92 g, 12.79 mmol) in THF (15. 99 ml) (drop by drop over ~5 minutes). After 1 hour, iodoethane (1.125 ml, 14.07 mmol) (neat) was added dropwise over ~5 minutes. The reaction was stirred for a further 2 hours at -78°C before slowly warming to room temperature. The reaction was then stirred overnight at room temperature. The reaction was quenched by pouring into 1:1 water/saline and extracting with EtOAc. The combined organics were washed with brine, dried with sodium sulfate, filtered and concentrated in vacuo. The crude residue was purified by silica gel column chromatography to provide Intermediate 83C (2.27 g, 8.86 mmol, 69% yield). TLC: product spots as purple dots in anisaldehyde (Rf = 0.80 in 1:1 Hex/EtOAc). 1H NMR (400 MHz, chloroform-d) δ: 4.14 (q, J=7.5 Hz, 2H), 3.88-3.95 (m, 4H), 2.09 (td, J =8.4, 5.6 Hz, 1H), 1.69-1.83 (m, 4H), 1.45-1.64 (m, 6H), 1.33-1.42 (m, 1H ), 1.25 (t, J=7.1 Hz, 3H), 0.86 (t, J=7.5 Hz, 3H) Intermediate 83D: ethyl 2-(4-oxocyclohexyl)butanoate

[00347] Intermediate 83C (2.00 g, 7.80 mmol) was taken up in THF (39.0 ml) and 1 M hydrochloric acid (39.0 ml) was added. The reaction is stirred at room temperature for 2 hours. The reaction was concentrated in vacuo, diluted with water and extracted with EtOAc. The combined organic extracts were dried with sodium sulfate, filtered and concentrated in vacuo. The crude material was purified over silica gel column chromatography to provide Intermediate 83D (1.47 g, 6.92 mmol, 89% yield). TLC: spots of faint pink product in anisaldehyde (Rf = 0.65 in 1:1 Hex/EtOAc). 1H NMR (400 MHz, chloroform-d) δ: 4.15 (q, J=7.1 Hz, 2H), 2.25-2.42 (m, 4H), 2.18 (ddd, J=9 .3, 7.8, 5.2 Hz, 1H), 2.10 (ddt, J=13.1, 6.2, 3.3 Hz, 1H), 1,902.03 (m, 2H), 1, 56-1.70 (m, 2H), 1.38-1.56 (m, 2H), 1.25 (t, J=7.2 Hz, 3H), 0.89 (t, J=7, 4 Hz, 3H) Intermediate 83E: Ethyl-2-((trans)-4-hydroxycyclohexyl)butanoate

[00348] Intermediate 83D (1.47 g, 6.92 mmol) was dissolved in EtOH (13.85 ml) and cooled to 0 °C. NaBH4 (0.314 g, 8.31 mmol) was added and the reaction was then allowed to stir at 0°C for 1 hour. The reaction was quenched with saturated aqueous NH4Cl and extracted with EtOAc. The combined organic extracts were dried with sodium sulfate, filtered, and concentrated in vacuo. The raw material was purified via silica gel column chromatography to provide Intermediate 83E (1.22 g, 5.69 mmol, 82% yield) along with (138 mg, 0.644 mmol, 9.30% yield) of the cis isomer. 1H NMR (400 MHz, chloroform-d) δ: 4.14 (q, J=7.1 Hz, 2H), 3.53 (t, J=10.5 Hz, 1H), 1.92-2, 08 (m, 2H), 1.80-1.89 (m, 1H), 1.63-1.70 (m, 1H), 1.52-1.62 (m, 4H), 1.37- 1.52 (m, 2H), 1.26 (t, J=7.2 Hz, 3H), 0.95-1.17 (m, 2H), 0.87 (t, J=7.4 Hz , 3H). Intermediate 83F:2-((trans)-4-((7-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)ethyl butanoate

[00349] Intermediate 83E (100 mg, 0.467 mmol) was taken up in DMSO (933 μl) and NaH (22.40 mg, 0.933 mmol) was added slowly, in portions at room temperature. After 1 hour, 4-chloro-7-(trifluoromethyl)quinoline (130 mg, 0.560 mmol) was added and the reaction was heated to 80 °C. After 16 hours, the reaction was quenched with ammonium chloride and extracted with EtOAc. The combined organic extracts were dried with sodium sulfate, filtered, and concentrated in vacuo. The crude residue was purified by silica gel column chromatography to provide Intermediate 83F (91 mg, 0.222 mmol, 47.6% yield). LC-MS Anal. Calculated for C22H26F3NO3 409.19, found [M+H] 410.2 Tr = 0.91 minutes (Method A). Intermediate 83G: 2-((trans)-4-((7-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)butanoic acid

[00350] Intermediate 83F (91 mg, 0.222 mmol) was captured in THF (889 μl), water (889 μl), and MeOH (445 μl). Lithium hydroxide (53.2 mg, 2.223 mmol) added and reaction stirred at 60°C for 40 hours. The reaction was concentrated in vacuo, diluted with water, acetic acid added (precipitate forms), the aqueous was extracted with EtO-Ac. The organics were combined, dried with sodium sulfate, filtered, and concentrated in vacuo to provide Intermediate 83G (85 mg, 0.223 mmol, 100% yield). LC-MS Anal. Calculated for C20H22F3NO3 381.16, found [M+H] 382.2 Tr = 0.78 minutes (Method A). Example 83: N-(4-Chlorophenyl)-2-((trans)-4-((7-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)butanamide

[00351] Intermediate 83G (41 mg, 0.108 mmol) was placed under a nitrogen atmosphere and trapped in SOCl2 (78 μl, 1.075 mmol). 1 drop of anhydrous DMF was added and the mixture was stirred for 1 hour at room temperature. The mixture was then concentrated in vacuo and co-evaporation with toluene in vacuo was used to remove the remaining thionyl chloride. Crude acyl chloride was dissolved in DCM (1075 μl) under a nitrogen atmosphere and TEA (74.9 μl, 0.538 mmol) was added followed by 4-chloroaniline (20.57 mg, 0.161 mmol). The mixture was stirred at room temperature for 30 minutes. The reaction was concentrated in vacuo, taken up in DMF, filtered, and purified by preparative HPLC to provide Example 83 (14.4 mg, 0.029 mmol, 27% yield). LC-MS Anal. Calculated for C26H26ClF3N2O2 490.16, found [M+H] 491.2 Tr = 0.94 minutes (Method A). 1H NMR (500 MHz, DMSO-d6) δ: 10.07 (s, 1H), 8.82 (d, J=5.1 Hz, 1H), 8.32 (d, J=8.6 Hz, 1H), 8.25 (s, 1H), 7.80 (d, J=8.7 Hz, 1H), 7.67 (d, J=8.5 Hz, 2H), 7.35 (d, J=8.5 Hz, 2H), 7.29 (d, J=5.0 Hz, 1H), 4.66 (t, J=10.1 Hz, 1H), 2.10-2.27 ( m, 3H), 1.97 (d, J=11.4 Hz, 1H), 1.73 (d, J=13.2 Hz, 1H), 1.41-1.65 (m, 5H), 1,281.41 (m, 1H), 1.21 (d, J=10.4 Hz, 1H), 0.85 (t, J=7.1 Hz, 3H). Enantiomer 1 and Enantiomer 2 Enantiomer 1: Example 83 (a) N-(4-Chlorophenyl)-2-((trans)-4-((7-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)- 2-butanamide (absolute stereochemistry not determined homochiral) Enantiomer 2: Example 83 (b) N-(4-Chlorophenyl)-2-((trans)-4-((7-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)-2-butanamide (stereochemistry absolute undetermined homochiral)

[00352] Example 83(a), Enantiomer 1, and Example 83(b), Enantiomer 2: Chiral separation of the racemic sample (Method C) gave Enantiomer 1 Tr = 3.611 minutes (Method D) and Enantiomer 2 Tr = 5.106 minutes (Method D). Absolute stereochemistry was not determined.

[00353] Example 83(a), Enantiomer 1: MS(ES): m/z = 491.1 [M+H]+. Tr = 2.529 minutes (Method B). 1H NMR (500 MHz, DMSO-d6) δ: 10.14 (s, 1H), 8.78 (d, J=5.0 Hz, 1H), 8.32 (d, J=8.6 Hz, 1H), 8.23 (s, 1H), 7.79 (d, J=8.8 Hz, 1H), 7.61 (d, J=8.8 Hz, 2H), 7.33 (d, J=8.7 Hz, 2H), 7.21 (d, J=5.3 Hz, 1H), 4.61 (br. s., 1H), 2.06-2.23 (m, 3H) , 1.94 (d, J=13.0 Hz, 1H), 1.69 (d, J=12.0 Hz, 1H), 1.36-1.62 (m, 5H), 1.30 ( d, J=13.2 Hz, 1H), 1.11-1.26 (m, 1H), 0.81 (t, J=7.1 Hz, 3H)

[00354] Example 83(b), Enantiomer 2: MS(ES): m/z = 491.1 [M+H]+. Tr = 2.545 minutes (Method B). 1H NMR (500 MHz, DMSO-d6) δ: 10.13 (s, 1H), 8.78 (d, J=5.1 Hz, 1H), 8.32 (d, J=8.7 Hz, 1H), 8.23 (s, 1H), 7.79 (d, J=8.7 Hz, 1H), 7.62 (d, J=8.8 Hz, 2H), 7.33 (d, J=8.8 Hz, 2H), 7.22 (d, J=5.2 Hz, 1H), 4.62 (br. s., 1H), 2.08-2.25 (m, 3H) , 1.95 (d, J=13.4 Hz, 1H), 1.70 (d, J=11.6 Hz, 1H), 1.39-1.62 (m, 5H), 1.30 ( d, J=12.3 Hz, 1H), 1.18 (d, J=11.4 Hz, 1H), 0.81 (t, J=7.1 Hz, 3H) Example 84 (±)-N -(4-Chlorophenyl)-2-((trans)-4-((2-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)butanamide Example 84: (±)-N-(4-Chlorophenyl)-2-((trans)-4-((2-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)butanamide

[00355] Example 84 was prepared from Intermediate 83E and the analogous procedures outlined to prepare 83F, 83G, and Example 83 except that 4-chloro-2-(trifluoromethyl)quinoline was used in part F. LC-MS Anal. Calculated for C26H26ClF3N2O2 490.16, found [M+H] 491.2 Tr = 1.20 minutes (Method A). 1H NMR (500 MHz, DMSO-d6) δ: 10.07 (s, 1H), 8.20 (d, J=8.2 Hz, 1H), 8.05 (d, J=8.7 Hz, 1H), 7.84-7.91 (m, 1H), 7.63-7.74 (m, 3H), 7.48 (s, 1H), 7.35 (d, J=8.7 Hz , 2H), 4,794.90 (m, 1H), 2.10-2.27 (m, 3H), 1.96 (d, J=13.1 Hz, 1H), 1.72 (d, J= 12.0 Hz, 1H), 1.44-1.63 (m, J=7.6 Hz, 5H), 1.32-1.42 (m, 1H), 1,201.32 (m, 1H), 0.85 (t, J=7.1 Hz, 3H). Example 85 (±)-N-(4-Chlorophenyl)-2-((trans)-4-(quinolin-4-yloxy)cyclohexyl)butanamide Example 85: (±)-N-(4-Chlorophenyl)-2-((trans)-4-(quinolin-4-yloxy)cyclohexyl)butanamide

[00356] Example 85 was prepared from Intermediate 83E and the analogous procedures outlined to prepare 83F, 83G, and Example 83 except that 4-bromo-quinoline was used in part F. LC-MS Anal. Calculated for C25H27ClN2O2 422.18, found [M+H] 423.2 Tr = 0.87 minutes (Method A). 1H NMR (500 MHz, DMSO-d6) δ: 10.14 (s, 1H), 8.65 (d, J=5.2 Hz, 1H), 8.11 (d, J=8.2 Hz, 1H), 7.91 (d, J=8.4 Hz, 1H), 7.72 (t, J=7.6 Hz, 1H), 7.63 (d, J=8.7 Hz, 2H) , 7.54 (t, J=7.5 Hz, 1H), 7.34 (d, J=8.6 Hz, 2H), 7.06 (d, J=5.2 Hz, 1H), 4 .58 (t, J=10.1 Hz, 1H), 2.08-2.26 (m, 3H), 1.95 (d, J=12.6 Hz, 1H), 1.70 (d, J=12.9 Hz, 1H), 1.39-1.66 (m, 5H), 1.31 (q, J=11.9 Hz, 1H), 1.10-1.25 (m, J =12.4 Hz, 1H), 0.82 (t, J=7.2 Hz, 3H). Example 86 N-(4-Chlorophenyl)-2-((trans)-4-((8-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)butanamide

[00357] Example 86 was prepared from Intermediate 83E and the analogous procedures outlined to prepare 83F, 83G, and Example 83 except that 4-chloro-8-(trifluoromethyl)quinoline was used in part F. LC-MS Anal. Calculated for C26H26ClF3N2O2 490.16, found [M+H] 491.2 Tr = 1.03 minutes (Method A). 1H NMR (500 MHz, DMSO-d6) δ: 10.13 (s, 1H), 8.77 (d, J=5.2 Hz, 1H), 8.39 (d, J=8.4 Hz, 1H), 8.12 (d, J=7.2 Hz, 1H), 7.58-7.67 (m, 3H), 7.33 (d, J=8.7 Hz, 2H), 7, 19 (d, J=5.4 Hz, 1H), 4.61 (t, J=10.4 Hz, 1H), 2.08-2.24 (m, 3H), 1.94 (d, J =12.0 Hz, 1H), 1.69 (d, J=13.8 Hz, 1H), 1.37-1.63 (m, 5H), 1.24-1.37 (m, J= 12.5 Hz, 1H), 1.11-1.24 (m, J=11.1 Hz, 1H), 0.81 (t, J=7.2 Hz, 3H) Example 87 Acid 2-(4 -(( R )-3-((4-chlorophenyl)amino)-2-(( cis )-4-(6-fluoroquinolin-4-yl)cyclohexyl)-3-oxopropyl)phenyl)propanoic acid Example 87: 2-(4-(( R )-3-((4-chlorophenyl)amino)-2-(( cis )-4-(6-fluoroquinolin-4-yl)cyclohexyl)-3-oxopropyl) acid phenyl)propanoic

[00358] Racemic example 87 can be prepared using General Procedures K with 4-chloroquiline, B, E, and L followed by alkylation with Intermediate 137A using the procedure for preparing 137B followed by the analogous procedures for intermediates 137C, D, and Example 137. Enantiomer 1 and Enantiomer 2 Enantiomer 1: Example 87(a) 2-(4-((R)-3-((4-chlorophenyl)amino)- 2-(( cis )-4-(6) acid -fluoroquinolin-4-yl)cyclohexyl)-3-oxopropyl)phenyl)-2-propanoic acid (stereochemistry not determined homochiral) Enantiomer 2: Example 87(b) 2-(4-((R)-3-((4-chlorophenyl)amino)- 2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl acid) -3-oxopropyl)phenyl)-2-propanoic (homochiral undetermined stereochemistry)

[00359] Example 87 (a) Enantiomer 1 and Example 87(b) Enantiomer 2: Chiral separation of the racemic sample (Method E) provided Enantiomer 1 Tr = 10.161 minutes (Method F) and Enantiomer 2 Tr = 13.160 minutes (Method F). Absolute stereochemistry was not determined.

[00360] Example 87 (a) Enantiomer 1: MS(ES): m/z = 541.3 [M+H]+. Tr = 0.84 minutes (Method A). 1H NMR (400 MHz, chloroform- d) δ: 8.78 (d, J=4.6 Hz, 1H), 8.09 (m, 2H), 7.69 (ddd, J=8.4, 7 .0, 1.2 Hz, 1H), 7.58 (ddd, J=8.4, 7.0, 1.1 Hz, 1H), 7.30 (d, J=4.5 Hz, 1H) , 7.11-7.20 (m, 8H), 6.91 (s, 1H), 3.72-3.78 (m, 1H), 3.02 (dd, J=13.2, 3, 4 Hz, 1H), 2.79-2.89 (m, J=13.0 Hz, 1H), 2.62 (td, J=10.8, 3.5 Hz, 1H), 2.28- 2.37 (m, 1H), 1.71-2.15 (m, 9H), 1.40 (d, J=7.1 Hz, 3H)

[00361] Example 87(b) Enantiomer 2: MS(ES): m/z = 541.3 [M+H]+. Tr = 0.84 minutes (Method A). 1H NMR (400 MHz, chloroform- d) δ: 8.82 (d, J=4.6 Hz, 1H), 8.05-8.14 (m, 2H), 7.70 (ddd, J=8 .3, 7.0, 1.2 Hz, 1H), 7.58 (ddd, J=8.3, 6.9, 1.2 Hz, 1H), 7.33 (d, J=4.6 Hz, 1H), 7.09-7.20 (m, 8H), 6.78 (s, 1H), 3.72-3.77 (m, 1H), 3.02 (dd, J=13, 1, 3.5 Hz, 1H), 2.83 (t, J=12.2 Hz, 1H), 2.61 (td, J=10.9, 3.5 Hz, 1H), 2.29- 2.37 (m, 1H), 1.72-2.16 (m, 9H), 1.40 (d, J=7.2 Hz, 3H) Example 88 (±)-N-(4-Chlorophenyl) -2-((cis)-4-((2-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)butanamide Intermediate 88A:2-((cis)-4-((2-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)ethyl butanoate

[00362] Intermediate 83E (300 mg, 1,400 mmol) was dissolved in THF (5600 μl) and 2-(trifluoromethyl)quinolin-4-ol (656 mg, 3.08 mmol) and triphenylphosphine (808 mg, 3.08 mmol ) were added. The solution was cooled to 0 °C in an ice bath. Diisopropyl azodicarboxylate (599 μl, 3.08 mmol) was added and the reaction was allowed to stir at room temperature once the addition was complete. Stirred at room temperature overnight. Next, the reaction was concentrated in vacuum and purified via silica gel column chromatography to provide Intermediate 88A (383 mg, 0.935 mmol, 66.8% yield). LC-MS Anal. Calculated for C22H26F3NO3 409.19, found [M+H] 410.2 Tr = 1.22 minutes (Method A). Intermediate 88B: 2-((cis)-4-((2-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)butanoic acid

[00363] Intermediate 88A (383 mg, 0.935 mmol) was captured in THF (748 μl), water (748 μl), and MeOH (374 μl). Lithium hydroxide (224 mg, 9.35 mmol) added and the reaction stirred at 60 °C overnight. After 16 hours, more lithium hydroxide (224 mg, 9.35 mmol) was added and the reaction heated for another 24 hours. The reaction was concentrated in vacuo, diluted with water, acidified with AcOH and extracted with EtOAc. The aqueous layer was extracted again with 7:3 chloroform:propanol. The combined organic extracts were dried with sodium sulfate, filtered, and concentrated in vacuum to provide Intermediate 88B (348 mg, 0.912 mmol, 98% production). LC-MS Anal. Calculated for C20H22F3NO3 381.16, found [M+H] 382.3 Tr = 1.04 minutes (Method A). Example 88: N-(4-Chlorophenyl)-2-((cis)-4-((2-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)butanamide

[00364] Intermediate 88B (50 mg, 0.131 mmol) was placed under a nitrogen atmosphere and trapped in SOCl2 (96 μl, 1.311 mmol). 1 drop of anhydrous DMF was added and the mixture was stirred for 1 hour at room temperature. The mixture was then concentrated in vacuo and co-evaporation with toluene in vacuo was used to remove the remaining thionyl chloride. Crude acyl chloride was dissolved in DCM (1311 μl) under a nitrogen atmosphere and TEA (91 μl, 0.655 mmol) was added followed by 4-chloroaniline (25.09 mg, 0.197 mmol). The mixture was stirred at room temperature for 30 minutes. The reaction was concentrated in vacuo, taken up in DMF, filtered, and purified by preparative HPLC to provide Example 88 (38.0 mg, 0.077 mmol, 59%). LC-MS Anal. Calculated for C26H26ClF3N2O2 490.16, found [M+H] 491.2 Tr = 1.18 minutes (Method A). 1H NMR (500 MHz, DMSO-d6) δ: 10.09 (s, 1H), 8.21 (d, J=8.2 Hz, 1H), 8.05 (d, J=8.5 Hz, 1H), 7.87 (t, J=7.5 Hz, 1H), 7.66 (t, J=7.6 Hz, 1H), 7.61 (d, J=8.8 Hz, 2H) , 7.36 (s, 1H), 7.32 (d, J=8.7 Hz, 2H), 5.13 (br. s., 1H), 2.17 (br. s., 1H), 2.03 (br. s., 2H), 1,601.78 (m, 4H), 1.46-1.60 (m, 4H), 1.34-1.46 (m, 1H), 0.81 (t, J=7.2 Hz, 3H) Enantiomer 1 and Enantiomer 2 Enantiomer 1: Example 88 (a) N-(4-Chlorophenyl)-2-(( cis )-4-((2- (trifluoromethyl)quinolin -4-yl)oxy)cyclohexyl)butanamide (absolute stereochemistry not determined homochiral) Enantiomer 2: Example 88(b) N-(4-Chlorophenyl)-2-(( cis )-4-((2- (trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)butanamide (ab-solute stereochemistry not determined homochiral)

[00365] Example 88(a) Enantiomer 1 and Example 88(b) Enantiomer 2: Chiral separation of the racemic sample (Method G) provided Enantiomer 1 Tr = 3.911 minutes (Method H) and Enantiomer 2 Tr = 4.551 minutes (Method H). Absolute stereochemistry was not determined.

[00366] Example 88(a), Enantiomer 1: MS(ES): m/z = 491.3 [M+H]+. Tr = 2.549 minutes (Method B). 1H NMR (500 MHz, DMSO-d6) δ: 10.05 (s, 1H), 8.22 (d, J=8.2 Hz, 1H), 8.06 (d, J=8.4 Hz, 1H), 7.88 (t, J=7.5 Hz, 1H), 7.68 (t, J=7.5 Hz, 1H), 7.63 (d, J=8.7 Hz, 2H) , 7.38 (s, 1H), 7.33 (d, J=8.7 Hz, 2H), 5.15 (br. s., 1H), 2.19 (br. s., 1H), 2.04 (d, J=7.1 Hz, 2H), 1.35-1.78 (m, 9H), 0.83 (t, J=7.2 Hz, 3H)

[00367] Example 88(b), Enantiomer 2: MS(ES): m/z = 491.3 [M+H]+. Tr = 2.541 minutes (Method B). 1H NMR (500 MHz, DMSO-d6) δ: 10.05 (s, 1H), 8.21 (d, J=8.2 Hz, 1H), 8.06 (d, J=8.5 Hz, 1H), 7.87 (t, J=7.5 Hz, 1H), 7.67 (t, J=7.6 Hz, 1H), 7.63 (d, J=8.7 Hz, 2H) , 7.37 (s, 1H), 7.33 (d, J=8.7 Hz, 2H), 5.15 (br. s., 1H), 2.18 (br. s., 1H), 1,982.10 (m, J=5.0 Hz, 2H), 1.36-1.80 (m, 9H), 0.83 (t, J=7.2 Hz, 3H) Example 89 (±)- N-(4-Chlorophenyl)-2-((cis)-4-((6-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)butanamide Example 89: (±)-N-(4-Chlorophenyl)-2-((cis)-4-((6-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)butanamide

[00368] Example 89 was prepared from Intermediate 83E and the analogous procedures outlined to prepare 88F, 88G, and Example 88 except that 4-hydroxy-6-(trifluoromethyl)quinoline was used in part F. LC-MS Anal. Calculated for C26H26ClF3N2O2 490.16, found [M+H] 491.3 Tr = 0.90 minutes (Method A). 1H NMR (500 MHz, DMSO-d6) δ: 10.06 (s, 1H), 8.83 (d, J=5.2 Hz, 1H), 8.43 (s, 1H), 8.14 ( d, J=8.8 Hz, 1H), 8.00 (d, J=8.2 Hz, 1H), 7.62 (d, J=8.7 Hz, 2H), 7.30 (d, J=8.7 Hz, 2H), 7.17 (d, J=5.3 Hz, 1H), 4.98 (br. s., 1H), 2.22 (br. s., 1H), 2.05 (br. s., 2H), 1.72 (d, J=13.1 Hz, 4H), 1.46-1.62 (m, 4H), 1.41 (d, J=11 .5 Hz, 1H), 0.83 (t, J=7.2 Hz, 3H) Enantiomer 1 and Enantiomer 2 Enantiomer 1: Example 89(a) N-(4-Chlorophenyl)-2-(( cis )- 4-((6-(trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)butanamide (absolute stereochemistry not determined homochiral) Enantiomer 2: Example 89(b) N-(4-Chlorophenyl)-2-(( cis )-4-((6- (trifluoromethyl)quinolin-4-yl)oxy)cyclohexyl)butanamide (ab-solute stereochemistry not determined homochiral)

[00369] Example 89(a) Enantiomer 1 and Example 89(b) Enantiomer 2: Chiral separation of the racemic sample (Method I) provided Enantiomer 1 Tr = 6.320 minutes (Method J) and Enantiomer 2 Tr = 7.500 minutes (Method J). Absolute stereochemistry was not determined.

[00370] Example 89(a), Enantiomer 1: MS(ES): m/z = 491.3 [M+H]+. Tr = 2.418 minutes (Method B). 1H NMR (500 MHz, DMSO-d6) δ: 10.05 (s, 1H), 8.82 (d, J=5.3 Hz, 1H), 8.43 (s, 1H), 8.13 ( d, J=8.8 Hz, 1H), 7.99 (d, J=8.8 Hz, 1H), 7.61 (d, J=8.8 Hz, 2H), 7.29 (d, J=8.8 Hz, 2H), 7.16 (d, J=5.4 Hz, 1H), 4.97 (br. s., 1H), 2.22 (br. s., 1H), 2.04 (br. s., 2H), 1.34-1.79 (m, 9H), 0.83 (t, J=7.2 Hz, 3H)

[00371] Example 89(b), Enantiomer 2: MS(ES): m/z = 491.3 [M+H]+. Tr = 2.418 minutes (Method B). 1H NMR (500 MHz, DMSO-d6) δ: 10.06 (s, 1H), 8.84 (d, J=5.2 Hz, 1H), 8.44 (s, 1H), 8.14 ( d, J=8.8 Hz, 1H), 7.99 (d, J=7.6 Hz, 1H), 7.63 (d, J=8.8 Hz, 2H), 7.30 (d, J=8.8 Hz, 2H), 7.17 (d, J=5.3 Hz, 1H), 4.98 (br. s., 1H), 2.23 (br. s., 1H), 2,012.11 (m, J=5.4 Hz, 2H), 1.37-1.79 (m, 9H), 0.84 (t, J=7.2 Hz, 3H). Example 90 N-(4-Chlorophenyl)-2-(4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)acetamide Intermediate 90A. tert-butyl 4-((4-chloroquinolin-6-yl)oxy)piperidine-1-carboxylate, and Intermediate 90B. tert-butyl 4-((6-fluoroquinolin-4-yl)oxy)piperidine-1-carboxylate

[00372] A solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (0.242 g, 1.200 mmol) in 1 mL of dioxane was treated with potassium hexamethyldisilazide (1.200 mL, 1.200 mmol) in THF. The resulting cloudy solution was stirred 5 min. at room temperature, then treated with 4-chloro-6-fluoroquinoline (0.182 g, 1 mmol) in 1 mL of dioxane. The reaction was brought to 60°C and stirred for 30 min, then overnight at room temperature. The reaction was quenched with 0.1 mL of glacial HOAc and subjected to silica gel chromatography (3:1 dichloromethane-EtOAc, then EtOAc, then 95:5 EtOAc-EtOH). Concentration of the appropriate fractions (high Rf) gave Intermediate 90A: tert-butyl 4-((4-chloroquinolin-6-yl)oxy)piperidine-1-carboxylate (0.11 g, 30% yield) as a colorless oil . MS(ES): m/z = 363 [M+H]+. tR = 0.93 minutes (Method A). Concentration of the low Rf fractions provided Intermediate 90B: tert-butyl 4-((6-fluoroquinolin-4-yl)oxy)piperidine-1-carboxylate (0.09 g, 26% yield) as a light yellow oil. MS(ES): m/z = 347 [M+H]+. tR = 0.77 minutes (Method A). Intermediate 90C. 4-Chloro-6-(piperidin-4-yloxy)quinoline

[00373] The mixture of Intermediate 90A (0.1 g, 0.276 mmol) in HCl/dioxane (1.033 ml, 4.13 mmol) was stirred at RT. The starting material lubricated on the glass after adding HCl. It still did not dissolve after sonication, so ~0.5 mL of dichloromethane was added, and this mixture was stirred for 3 hours. During this time, dissolution of the oil appears to have occurred, and a white precipitate formed. The reaction was pumped to dryness to provide 0.08 g (97%) of Intermediate 90C, HCl as an off-white powder. MS(ES): m/z = 263 [M+H]+. tR = 0.50 minutes (Method A). Intermediate 90D. Ethyl 2-(4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)acetate

[00374] A suspension of 4-chloro-6-(piperidin-4-yloxy)quinoline, HCl (0.06 g, 0.201 mmol) and potassium carbonate (0.097 g, 0.702 mmol) in DMF (0.5 mL) was treated with ethyl bromoacetate (0.045 mL, 0.401 mmol), and the resulting mixture was stirred 4 hours at room temperature. This mixture was neutralized with glacial HOAc, and the residue was purified by flash chromatography (1:1 EtOAc-CH2Cl2 to EtOAc). Concentration of appropriate fractions of Intermediate 90D as a colorless oil. MS(ES): m/z = 349 [M+H]+. tR = 0.56 minutes (Method A). Intermediate 90E. 2-(4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)acetic acid, 2 HCl

[00375] A solution of 0.04 g of Intermediate 90D in THF (2 mL) was treated with lithium hydroxide (0.04 g, 1.670 mmol) in water (1 mL). Methanol, ~1 mL, was added to provide a single phase, and the reaction was stirred 2 hours at room temperature. The solvent was removed under a stream of nitrogen, and the resulting suspension was treated with ~3 mL of water to dissolve. Aqueous HCl was added slowly, eventually bringing the solution pH to ~4, but the product did not precipitate at any point. The pH was brought back to ~7.5 with aqueous sodium bicarbonate. Brine has been added but has not yet precipitated. This solution was ext. six times with 3:1 chloroform-IPA, and the combined organic extracts were stripped to provide an oil. This was treated with 5 eq. of HCl/dioxane, and the drop of water was added to dissolve. This solution was then lyophilized to provide Intermediate 90E (0.04 g, 92%) as a greenish-brown solid. MS(ES): m/z = 321 [M+H]+. tR = 0.48 minutes (Method A). Example 90. N-(4-Chlorophenyl)-2-(4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)acetamide

[00376] A solution of Intermediate 90E (0.04 g, 0.102 mmol) and 4-chloroaniline (0.014 g, 0.112 mmol) and triethylamine (0.057 mL, 0.406 mmol) in DMF (1 mL) was treated with BOP (0.067 g , 0.152 mmol). This solution was stirred 2 hours at RT, then purified by preparative HPLC. Concentration of the appropriate fraction provided material with a purity of ~90%. This material was also purified by flash chromatography. Concentration of the appropriate fractions, then lyophilization provided Example 90 (0.016 g, 36% yield) as a white powder. MS(ES): m/z = 430 [M+H]+. tR = 0.73 minutes (Method A). 1H NMR (400 MHz, DMSO-d6) δ 9.88 (s, 1H), 8.69 (d, 1H, J = 4.7 Hz), 8.04 (d, 1H, J = 9.2 Hz ), 7.69-7.74 (m, 3H), 7.57 (dd, 1H, J = 9.2, 2.8 Hz), 7.50 (d, 1H, J = 2.7 Hz) , 7.36-7.39 (m, 2H), 4.69-4.76 (m, 1H), 3.19 (s, 2H), 2.78-2.86 (m, 2H), 2 .06-2.13 (m, 2H), 1.81-1.89 (m, 2H). Note: a signal at ~2.55 ppm is largely obscured by solvent. Example 91 (±)-N-(4-Chlorophenyl)-2-(4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)propanamide Intermediate 91A.2-(±)-ethyl4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)propanoate

[00377] (±)-ethyl 2-(4-((4-Chloroquinolin-6-yl)oxy)piperidin-1-yl)propanoate was prepared as a light yellow oil at 97% production at 90C and 2- ethyl bromopropionate using the procedure for the conversion of 90C to 90D except that the reaction was carried out at 60 °C and no chromatography was performed. MS(ES): m/z = 363 [M+H]+. tR = 0.58 minutes (Method A). Intermediate 91B. (±)-2-(4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)propanoic acid

[00378] (±)-2-(4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)propanoic acid was prepared as an off-white foam at 93% production of 91A using the procedure for the conversion of 90D to 90E except that the isolated material was not converted to the HCl salt. MS(ES): m/z = 335 [M+H]+. tR = 0.49 minutes (Method A). Example 91. (±)-N-(4-Chlorophenyl)-2-(4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)propanamide

[00379] (±)-N-(4-Chlorophenyl)-2-(4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)propanamide was prepared as the trifluoroacetate salt in 57% production after preparative HPLC purification of 91B using the procedure for converting 90E to Example 90. MS(ES): m/z = 444 [M+H]+. tR = 0.73 minutes (Method A). 1H NMR (400 MHz, DMSO-d6) δ 10.83 (s, 1H), 8.69 (d, 1H, J = 4.4 Hz), 8.07 (d, 1H, J = 9.1 Hz ), 7.73 (d, 1H, J = 4.2 Hz), 7.59-7.65 (m, 3H), 7.56 (s, 1H), 7.49 (d, 2H, J = 8.1 Hz), 4.63-4.67 (m, 1H), 3.10-3.70 (m, 5H), 2.01-2.39 (m, 4H), 1.66 (d , 3H, J = 6.7 Hz). Example 92 (±)-2-(4-((4-Chloroquinolin-6-yl)oxy)piperidin-1-yl)-N-(p-tolyl)propanamide Example 92. (±)-2-(4-((4-Chloroquinolin-6-yl)oxy)piperidin-1-yl)-N-(p-tolyl)propanamide

[00380] (±)-2-(4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)-N-(p-tolyl)propanamide was prepared as the HCl salt in 77% production after purification by preparative HPLC and salt switching of 91B and p-toluidine using the procedure for converting 90E to Example 90. MS(ES): m/z = 424 [M+H]+. tR = 0.71 minutes (Method A). 1H NMR (400MHz, DMSO-d6) δ 11.02 (s, not integrated), 10.91 (s, not integrated), 10.72 (br. s, not integrated), 10.57 (br. s, unintegrated), 8.78 (s, 1H), 8.13 (t, 1H, J = 9.8 Hz), 7.83 (s, 1H), 7.52-7.70 (m, 4H) , 7.19 (d, 2H, J = 7.2 Hz), 4.27-4.35 (m, 1H), 3.25-3.77 (m, 5H), 1,962.41 (m, 7H ), 1.62 (d, 3H, J = 5.1 Hz). Enantiomer 1 and Enantiomer 2 of Racemic Example 91 Enantiomer 1: Example 93 N-(4-Chlorophenyl)-2-(4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)propanamide (Homochiral absolute stereochemistry was not determined) Enantiomer 2: Example 94 N-(4-Chlorophenyl)-2-(4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)propanamide (Absolute homochiral stereochemistry was not determined) Examples 93 and 94. N-(4-Chlorophenyl)-2-(4-((4-chloroquinolin-6-yl)oxy)piperidin-1-yl)propanamide (both enantiomers, absolute stereochemistry not determined)

[00381] The material from racemic Example 91 (0.038 g) was purified by chiral SFC (27% MeOH in CO2, 0.1% (v/v) each of diethylamine and ammonium formate, CHIRALPAK® AD column, 85 ml /min). Concentration of the appropriate fraction (previous elution) provided:

[00382] Example 93 (enantiomer 1, first elution, absolute stereochemistry was not determined) (0.007 g, 23%). MS(ESI): m/z = 444 [M+H]+. tR = 1.28 minutes (SCP TFA). 1H NMR (400MHz, DMSO-d6) δ 8.65 (d, 1H, J = 4.5 Hz), 8.02 (d, 1H, J = 9.2 Hz), 7.69 (d, 1H, J = 4.5 Hz), 7.64 (d, 2H, J = 8.6 Hz), 7.54 (br. d, 1H, J = 9.3 Hz), 7.47 (br. s, 1H), 7.37 (d, 2H, J = 8.5 Hz), 4.68-4.75 (m, 1H), 3.56-3.0 (m, 5H), 2.06-2 .15 (m, 2H), 1.80-1.90 (m, 2H), 1.24-1.32 (m, 3H).

[00383] Example 94 (enantiomer 2, second elution, absolute stereochemistry was not determined) (0.027 g). MS(ESI): m/z = 444 [M+H]+. tR = 1.27 minutes (Method K). 1H NMR (400MHz, DMSO-d6): essentially the same as racemate NMR (some ammonium diethylformate is present). Example 95 N-(4-Chlorophenyl)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)acetamide Intermediate 95A. 6-Fluoro-4-(piperidin-4-yloxy)quinoline, 2 HCl

[00384] 6-Fluoro-4-(piperidin-4-yloxy)quinoline, 2 HCl was prepared from 90B in quantitative production as a brown solid using the conditions for the conversion of 90A to 90C. MS(ES): m/z = 247 [M+H]+. tR = 0.42 minutes (Method A). Intermediate 95B. Ethyl 2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)acetate

[00385] Ethyl 2-(4-((6-Fluoroquinolin-4-yl)oxy)piperidin-1-yl)acetate was prepared at 63% production as a 95A light yellow oil using the conditions for the conversion of 90C to 90D. MS(ES): m/z = 333 [M+H]+. tR = 0.48 minutes (Method A). Intermediate 95C. 2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)acetic acid

[00386] 2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)acetic acid was prepared in 75% production as a yellow glass of 95B using the conditions for the conversion of 90D to 90E except that the isolated material was not converted to the HCl salt. MS(ES): m/z = 305 [M+H]+. tR = 0.42 minutes (Method A). Example 95. N-(4-Chlorophenyl)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)acetamide

[00387] N-(4-chlorophenyl)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)acetamide was prepared in 38% production at 95C using the conditions for the conversion from 90E to Example 90. MS(ES): m/z = 414 [M+H]+. tR = 0.66 minutes (Method A). 1H NMR (400 MHz, DMSO-d6) δ 8.68 (d, 1H, J = 5.1 Hz), 8.04 (dd, 1H, J = 9.0, 5.2 Hz), 7.81 (d, 1H, J = 9.3 Hz), 7.63-7.68 (m, 3H), 7.36 (d, 2H, J = 8.4 Hz), 7.13 (d, 1H, J = 5.0 Hz), 4.81-4.87 (m, 1H), 3.69 (s, 2H), 2.80-2.88 (m, 2H), 2.58-2.67 (m, 2H), 2.07-2.14 (m, 2H), 1.89-1.98 (m, 2H). Example 96 (±)-N-(4-Chlorophenyl)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)propanamide Intermediate 96A.2-(±)-ethyl4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)propanoate

[00388] (±)-ethyl 2-(4-((6-Fluoroquinolin-4-yl)oxy)piperidin-1-yl)propanoate was prepared as a colorless oil in 83% production of 95A and 2-bromopropionate of ethyl using the procedure for converting 90C to 90D. MS(ES): m/z = 347 [M+H]+. tR = 0.51 min. (Method A). Intermediate 96B. (±)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)propanoic acid

[00389] (±)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)propanoic acid was prepared as a glass in 68% production of 96A using the procedure for conversion from 90D to 90E except that the isolated material was not converted to the HCl salt. MS(ES): m/z = 319 [M+H]+. tR = 0.42 min. (Method A). Example 96.(±)-N-(4-Chlorophenyl)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)propanamide

[00390] (±)-N-(4-chlorophenyl)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)propanamide was prepared at 58% yield after purification by HPLC preparative of 96B using the procedure for converting 90E to Example 90. MS(ES): m/z = 428 [M+H]+. tR = 0.72 minutes (Method A). 1H NMR (400 MHz, DMSO-d6) δ 9.97 (s, 1H), 8.69 (d, 1H, J = 5.4 Hz), 8.02 (dd, 1H, J = 9.8, 5.4 Hz), 7.77 (dd, 1H, J = 9.7, 2.8 Hz), 7.71 (d, 2H, J = 8.1 Hz), 7.64-7.68 ( m, 1H), 7.37 (d, 2H, J = 8.8 Hz), 7.15 (d, 1H, J = 5.2 Hz), 4.78-4.85 (m, 1H), 2.77-2.87 (m, 2H), 2.57-2.63 (m, 1H), 2.06-2.14 (m, 2H), 1.85-1.94 (m, 2H ), 1.22 (d, 3H, J = 6.8 Hz). Note: Signal is largely obscured by solvent. Examples 97 to 101 Intermediate 97A: (±)-ethyl 2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)butanoate

[00391] (±)-ethyl 2-(4-((6-Fluoroquinolin-4-yl)oxy)piperidin-1-yl)butanoate was prepared as a colorless oil in 71% production of 95A and 2-bromobutyrate of ethyl using the procedure for the conversion of 90C to 90D except that the reaction was carried out at 50 °C. MS(ES): m/z = 361 [M+H]+. tR = 0.54 min. (Method A). Intermediate 97B: (±)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)butanoic acid

[00392] (±)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)butanoic acid was prepared as a straw-colored foam at 74% 97A production using the procedure for the conversion of 90D to 90E except that the isolated material was not converted to the HCl salt. MS(ES): m/z = 333 [M+H]+. tR = 0.44 min. (Method A). Examples 97 to 101:

[00393] Bop coupling (Scheme 1, below) of carboxylic acids x (Intermediates 96B and 97B prepared in the previous examples) with the appropriate anilines under the conditions described for the conversion of 90E to Example 90 provides compounds of invention 1 shown in table 2 below. (All entries are racemic) Scheme 1 Table 2 Example 102 (±)-N-(4-Chlorophenyl)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)-3-methylbutanamide 102A. (±)-ethyl 2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)-3-methylbutanoate

[00394] (±)-ethyl 2-(4-((6-Fluoroquinolin-4-yl)oxy)piperidin-1-yl)-3-methylbutanoate was prepared as a light yellow oil at 59% production of 95A and ethyl 2-bromo-3-methylbutyrate using the procedure for converting 90C to 90D except that the reaction was carried out at 90 °C. MS(ES): m/z = 375 [M+H]+. tR = 0.60 min. (Method A). 102B. (±)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)-3-methylbutanoic acid

[00395] (±)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)-3-methylbutanoic acid was prepared as an off-white solid at 77% production of 102A using the procedure for converting 90D to 90E except that the reaction was carried out for several days at 75 ° C and the isolated material was not converted to the HCl salt. MS(ES): m/z = 347 [M+H]+. tR = 0.47 min. (Method A). Example 102.(±)-N-(4-Chlorophenyl)-2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)-3-methylbutanamide

[00396] A solution of 2-(4-((6-fluoroquinolin-4-yl)oxy)piperidin-1-yl)-3-methylbutanoic acid (0.02 g, 0.058 mmol) and N-methylmorpholine (0.013 mL , 0.115 mmol) in THF (0.2 mL) was cooled to 0 °C and treated with isobutyl chloroformate (9.10 μL, 0.069 mmol). This mixture was stirred 15 min, then treated with 4-chloroaniline (8.84 mg, 0.069 mmol) and warmed to room temperature and stirred 1 h. The reaction was quenched by adding 1 drop of water, diluted with DMF, and purified by preparative HPLC. Concentration of the appropriate fraction provided Example 102, 2 TFA (0.005 g, 13% yield) as a white powder. MS(ES): m/z = 456 [M+H]+. tR = 0.82 minutes (Method A). 1H NMR (400 MHz, DMSO-d6) δ 10.94 (br. s, 1H), 9.96 (br. s, 1H), 8.02-8.19 (m, 3H), 7.82- 7.91 (m, 1H), 7.67 (d, 2H, J = 8.7 Hz), 7.44 (d, 2H, J = 8.2 Hz), 2.01-2.37 (m , 5H), 1.10 (d, 6H, J = 5.3 Hz). Note: Several obscure signs due to a large water peak. Examples 103 to 112

[00397] Chiral SFC (Scheme 2, below) of racemic materials (prepared in the previous examples) with the indicated columns under the conditions (MeOH-CO2) described for the resolution of Example 91 into its component enantiomers of Example 93 and Example 94 provides homochiral compounds of invention 1 shown in table 2 below. (All inputs are homochiral undetermined absolute stereochemistry) Scheme 2 Table 3 Example 113 N-(4-Chlorophenyl)-2-(1-(quinolin-4-ylmethyl)piperidin-4-yl)acetamide Intermediate 113A. Methyl 2-(1-(quinolin-4-ylmethyl)piperidin-4-yl)acetate

[00398] A solution of methyl 2-(piperidin-4-yl)acetate, HCl (0.465 g, 2.400 mmol) and triethylamine (0.892 ml, 6.40 mmol) in 1 ml of DMF was treated with 4-(chloromethyl )quinoline, HCl (0.428 g, 2 mmol). The resulting solution was stirred at room temperature for 3 hours, then diluted with EtOAc. The organic extracts were then washed with water, dried, filtered, and stripped to provide methyl 2-(1-(quinolin-4-ylmethyl)piperidin-4-yl)acetate (0.54 g, 86% yield). like an amber oil. MS(ES): m/z = 299 [M+H]+. tR = 0.48 minutes (Method A). Intermediate 113B. 2-(1-(quinolin-4-ylmethyl)piperidin-4-yl)acetic acid

[00399] A solution of Intermediate 113A (0.1 g, 0.335 mmol) in THF (1 mL) was treated with aq. lithium hydroxide. (0.268 mL, 0.670 mmol) followed by 0.2 mL of water. Methanol, ∼0.5 mL, was added to provide a single phase, and the reaction was stirred 18 h at RT. Most of the organic solvent was removed under a stream of nitrogen, and the residue was quenched with 0.08 mL of glacial HOAc. The pH was brought to ~7 with saturated aqueous sodium bicarbonate, and the resulting solution was ext. three times with 3:1 chloroform-isopropanol. The combined organic extracts were dried, filtered, and stripped to provide 2-(1-(quinolin-4-ylmethyl)piperidin-4-yl)acetic acid (0.095 g, 95% yield) as a brown solid. MS(ES): m/z = 285 [M+H]+. tR = 0.43 minutes (Method A). Example 113. N-(4-Chlorophenyl)-2-(1-(quinolin-4-ylmethyl)piperidin-4-yl)acetamide

[00400] N-(4-Chlorophenyl)-2-(1-(quinolin-4-ylmethyl)piperidin-4-yl)acetamide was prepared in 51% production of 113B using the conditions for the conversion of 90E to Example 90 MS(ES): m/z = 394 [M+H]+. tR = 1.07 minutes (Method K). 1H NMR (400 MHz, DMSO-d6) δ 10.08 (s, 1H), 8.82 (d, 1H, J = 3.9 Hz), 8.27 (d, 1H, J = 8.3 Hz ), 8.02 (d, 1H, J = 8.3 Hz), 7.75 (t, 1H, J = 7.5 Hz), 7.55-7.64 (m, 3H), 7.49 (d, 1H, J = 3.4 Hz), 7.33 (d, 2H, J = 8.4 Hz), 3.76 (s, 2H), 2,812.88 (m, 2H), 2.22 (d, 2H, J = 6.7 Hz), 2.03-2.13 (m, 2H), 1.73-1.82 (m, 1H), 1.59-1.65 (m, 2H ), 1.19-1.28 (m, 2H). Example 114 N-(4-Fluorophenyl)-2-(1-(quinolin-4-ylmethyl)piperidin-4-yl)acetamide

[00401] N-(4-Fluorophenyl)-2-(1-(quinolin-4-ylmethyl)piperidin-4-yl)acetamide was prepared in 76% production of 113B and 4-fluoroaniline using the conditions for the conversion of 90E for Example 90. MS(ES): m/z = 378 [M+H]+. tR = 0.88 minutes (Method K). 1H NMR (400 MHz, DMSO-d6) δ 10.03 (s, 1H), 8.79 (d, 1H, J = 4.0 Hz), 8.25 (d, 1H, J = 8.3 Hz ), 8.01 (d, 1H, J = 8.4 Hz), 7.75 (t, 1H, J = 7.5 Hz), 7.61 (t, 1H, J = 7.5 Hz), 7.47-7.55 (m, 3H), 7.09 (t, 2H, J = 8.7 Hz), 3.90 (s, 2H), 2.80-2.86 (m, 2H) , 2.20 (d, 2H, J = 6.9 Hz), 2.03-2.11 (m, 2H), 1.71-1.79 (m, 1H), 1.58-1.64 (m, 2H), 1.18-1.27 (m, 2H). Example 115 N-(4-Fluorophenyl)-2-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)acetamide Intermediate 115A. Methyl 2-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)acetate

[00402] To the mixture of 4-chloro-6-fluoroquinoline (200.0 mg, 1.1 mmol) in anhydrous NMP (4 mL), in a sealable vial, was added 2-(piperidin-4-yl)acetate. methyl (260.0 mg, 1.7 mmol) followed by DIPEA (0.8 mL, 4.6 mmol). The vial was capped and the mixture was stirred at room temperature for two hours, then at 120°C for 66 hours. The reaction mixture was cooled to room temperature before being partitioned between water and Et2O. The layers were separated and the aqueous layer was extracted twice more with Et2O. These organic extracts were combined with the original organic layer and washed with brine, dried (Na2SO4), filtered and concentrated in vacuo to provide the crude product. Purification by Isco chromatography provided methyl 2-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)acetate as a golden oil (304.3 mg; 91% yield). MS(ES): m/z = 303 [M+H]+. tR = 0.64 minutes (Method A). 1H NMR (400 MHz, DMSO-d6) δ 8.67 (d, J=5.0 Hz, 1H), 8.04 - 7.97 (m, 1H), 7.67 - 7.53 (m, 2H), 7.02 (d, J=5.0 Hz, 1H), 3.63 (s, 3H), 3.51 - 3.43 (m, 2H), 2.87 - 2.76 (m , 2H), 2.39 (d, J=7.0 Hz, 2H), 1.94 - 1.89 (m, 1H), 1.87 - 1.81 (m, 2H), 1.62 - 1.49 (m, 2H). Intermediate 115B. 2-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)acetic acid

[00403] To a homogeneous mixture of Intermediate 115A (304.3 mg, 1.0 mmol) in EtOH (10 mL), at room temperature under a nitrogen atmosphere, 2 M NaOH (aq) was added dropwise (1 mL, 2.0 mmol). The mixture was stirred at room temperature for 22 hours before being quenched with 4M HCl in dioxane (0.5 mL, 2.0 mmol). After stirring for 5 minutes, the mixture was concentrated in vacuo to provide a pale gold solid, which was used in the next step without further purification. MS(ES): m/z = 289 [M+H]+. tR = 0.55 minutes (Method A). Example 115. N-(4-Fluorophenyl)-2-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)acetamide

[00404] To the mixture of Intermediate 115B (20.0 mg, 0.07 mmol) and 4-fluoroaniline (9.3 mg, 0.08 mmol) in anhydrous DMF (1 mL), at room temperature under nitrogen, DIPEA (0.04 mL, 0.23 mmol) was added followed by PyBOP (36.1 mg, 0.07 mmol). The resulting mixture was stirred at room temperature for 4 hours before being concentrated in vacuo to remove volatiles, diluted with DMSO, filtered through a syringe filter, then purified by preparative HPLC/MS to provide the title compound ( 14.4 mg; 54% production). MS(ES): m/z = 382 [M+H]+. tR = 1.25 minutes (Method K). 1H NMR (500 MHz, DMSO-d6) δ 10.04 (s, 1H), 8.66 (d, J=4.9 Hz, 1H), 8.03 - 7.99 (m, 1H), 7 .64 - 7.58 (m, 4H), 7.16 - 7.11 (m, 2H), 7.03 (d, J=4.9 Hz, 1H), 3.59 - 3.15 (m , 2H), 2.86 - 2.78 (m, 2H), 2.36 (d, J=7.1 Hz, 2H), 2.07 - 1.98 (m, 1H), 1.89 - 1.84 (m, 2H), 1.62 - 1.53 (m, 2H). Examples 116 to 119

[00405] Reaction of Intermediate 115B with an appropriate amine, under the conditions described by example 115 (Scheme 3, below), provides compounds of the invention shown in table 3 below. Scheme 3 Table 4 Example 120 N-(4-Fluorophenyl)-2-(1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)acetamide Intermediate 120A. Methyl 2-(4-methylpiperidin-4-yl)acetate, HCl

[00406] To a flask charged with MeOH (7.5 mL), at 0 °C under a nitrogen atmosphere, acetyl chloride (1.1 mL, 15.2 mmol) was slowly added. After the addition was complete, the mixture was stirred at 0°C for 5 minutes before a homogeneous mixture of 2-(4-methylpiperidin-4-yl)acetic acid, HCl (675.0 mg, 3.5 mmol) in MeOH (1.5 mL) will be added slowly dropwise. The resulting homogeneous mixture was stirred at 0°C for 5 minutes, then at 60°C for 8 hours, before being concentrated in vacuo to provide the HCl salt of Intermediate 120A as a white solid (718.0 mg; 99 % production) that was used without further purification. 1H NMR (400 MHz, DMSO-d6) δ 9.41 - 9.12 (m, 1H), 3.60 (s, 3H), 3.25 - 3.15 (m, 2H), 2.93 - 2.82 (m, 2H), 2.39 - 2.30 (m, 2H), 1.74 - 1.64 (m, 4H), 1.02 (s, 3H). Intermediate 120B. Methyl 2-(1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)acetate

[00407] To a homogeneous mixture of 4-chloro-6-fluoroquinoline (350.0 mg, 1.9 mmol) in anhydrous NMP (5 mL), in a sealable vial, was added the HCl salt of 2- Methyl (4-methylpiperidin-4-yl)acetate (120A, 480.0 mg, 2.3 mmol) followed by DIPEA (1.6 mL, 9.2 mmol). The vial was sealed and the mixture was stirred at 120°C. After 26 hours, the reaction mixture was cooled to room temperature, then partitioned between water and EtOAc. The layers were separated and the aqueous layer was extracted once again with EtOAc. The organic layers were combined, washed with brine, then concentrated in vacuo to provide the crude product. Purification by Isco chromatography provided Intermediate 120B as an oil (565.8 mg; 93% yield). MS(ES): m/z = 317 [M+H]+. tR = 0.66 minutes (Method A). 1H NMR (400 MHz, DMSO-d6) δ 8.38 (d, J=5.4 Hz, 1H), 7.96 (dd, J=11.7, 2.8 Hz, 1H), 7.89 - 7.84 (m, 1H), 7.55 - 7.49 (m, 1H), 6.54 (d, J=5.5 Hz, 1H), 3.82 - 3.63 (m, 2H ), 3.59 (s, 3H), 3.54 - 3.34 (m, 2H), 2.45 - 2.38 (m, 2H), 1.87 - 1.72 (m, 4H), 1.05 (s, 3H). Intermediate 120C. 2-(1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)acetic acid

[00408] To a homogeneous mixture of methyl 2-(1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)acetate (321.0 mg, 1.0 mmol) in MeOH (5 mL ), under a nitrogen atmosphere, 2 M NaOH aqueous solution (1 mL, 2.0 mmol) was added dropwise. The reaction was then stirred at room temperature for 20 hours before being treated with 1 N HCl (aq) to pH 6 on pH test strips. The mixture was then partitioned between water and EtOAc, the layers were separated and the aqueous layer was extracted twice with EtOAc. The aqueous extraction layer was lyophilized to provide crude Example 120C as an off-white solid (302.1 mg, 98% yield) which was used without further purification. MS(ES): m/z = 303 [M+H]+. tR = 0.59 minutes (Method A). 1H NMR (400 MHz, DMSO-d6) δ 12.12 (br.s, 1H), 8.41 (d, J=6.1 Hz, 1H), 8.12 (dd, J=11.5, 2.7 Hz, 1H), 8.00 (dd, J=9.3, 5.7 Hz, 1H), 7.75 - 7.64 (m, 1H), 6.64 (d, J=6 .2 Hz, 1H), 3.98 - 3.87 (m, 1H), 3.87 - 3.78 (m, 1H), 3.69 - 3.49 (m, 2H), 2.38 - 2.29 (m, 2H), 1.92 - 1.70 (m, 4H), 1.06 (s, 3H). Example 120.N-(4-Fluorophenyl)-2-(1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)acetamide

[00409] To the mixture of 2-(1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)acetic acid (25.6 mg, 0.09 mmol) and 4-fluoroaniline (11.3 mg, 0.1 mmol) in anhydrous DMF (1 mL), under nitrogen atmosphere, DIPEA (0.05 mL, 0.3 mmol) was added followed by PyBOP (44.1 mg, 0.09 mmol). The resulting mixture was stirred at room temperature for 156 hours, before being diluted with DMF, filtered through a syringe filter, then purified by preparative HPLC/MS to provide the title compound (17.2 mg; 51% of production). MS(ES): m/z = 396 [M+H]+. tR = 1.32 minutes (Method K). 1HKNMR (500 MHz, DMSO-d6) δ 10.05 (s, 1H), 8.33 (d, J=5.5 Hz, 1H), 7.99 - 7.93 (m, 1H), 7, 89 - 7.83 (m, 1H), 7.57 - 7.50 (m, 3H), 7.13 - 7.08 (m, 2H), 6.54 (d, J=5.5 Hz, 1H), 3.85 - 3.61 (m, 2H), 3.59 - 3.36 (m, 2H), 2.41 - 2.31 (m, 2H), 1.86 - 1.75 ( m, 4H), 1.07 (s, 3H). Examples 121 to 125

[00410] Reaction of 2-(1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)acetic acid with an appropriate amine, under the conditions described for example 120 (Scheme 4, below), provides compounds of the invention shown in table 4 below. Scheme 4 Table 5 Example 126 (±)-N-(4-Chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)propanamide Intermediate 126A. tert-butyl 4-(6-fluoroquinolin-4-yl)piperazine-1-carboxylate

[00411] To the mixture of 4-chloro-6-fluoroquinoline (200.0 mg, 1.1 mmol) in anhydrous NMP (4 mL), in a sealable vial, was added 1-Boc-piperizine (308.0 mg, 1.7 mmol) followed by DIPEA (0.8 mL, 4.6 mmol). The vial was capped and the mixture was stirred at room temperature for two hours, then at 120°C for 66 hours. The reaction mixture was cooled to room temperature before being partitioned between water and Et2O. The layers were separated and the aqueous layer was extracted twice more with Et2O. These organic extracts were combined with the original organic layer and washed with brine, dried (Na2SO4), filtered and concentrated in vacuo to provide the crude product. Purification by Isco chromatography provided tert-butyl 4-(6-fluoroquinolin-4-yl)piperazine-1-carboxylate as an oil (362.8 mg; 98% yield). MS(ES): m/z = 332 [M+H]+. tR = 0.70 minutes (Method A). 1H NMR (400 MHz, DMSO-d6) δ 8.70 (d, J=4.9 Hz, 1H), 8.09 - 8.00 (m, 1H), 7.77 - 7.67 (m, 1H), 7.67 - 7.62 (m, 1H), 7.07 (d, J=4.9 Hz, 1H), 3.67 - 3.57 (m, 4H), 3.15 - 3 .06 (m, 4H), 1.44 (s, 9H). Intermediate 126B. 6-Fluoro-4-(piperazin-1-yl)quinoline

[00412] To a flask loaded with tert-butyl 4-(6-fluoroquinolin-4-yl)piperazine-1-carboxylate (362.8 mg, 1.1 mmol), under a nitrogen atmosphere, HCl was added at 4 M in dioxane (10 mL, 40.0 mmol). The resulting mixture was stirred at room temperature for 5.5 hours, during which time a precipitate formed. The heterogeneous mixture was concentrated to approximately 1/2 of its original volume. Vacuum filtration provided the HCl salt of the title compound as an off-white solid (259.0 mg; 88% yield) which was used without further purification. MS(ES): m/z = 232 [M+H]+. tR = 0.34 minutes (Method A). Intermediate 126C. Ethyl 2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)propanoate

[00413] A heterogeneous mixture of 6-fluoro-4-(piperazin-1-yl)quinoline HCl salt (126B, 80.0 mg, 0.3 mmol) in anhydrous NMP (2 mL), in a vial sealable, K2CO3 (60.0 mg, 0.4 mmol) was added followed by ethyl 2-bromopropanoate (65.0 mg, 0.4 mmol). The vial was then sealed and the mixture was stirred at 60°C. After 67.5 hours, the reaction was cooled to room temperature, then partitioned between water and DCM. The layers were separated and the aqueous layer was extracted once again with DCM. The organic layers were combined and concentrated in vacuo to provide the product as an oil (92.7 mg, 94%), which was used without further purification. MS(ES): m/z = 332 [M+H]+. tR = 0.51 minutes (Method A). Intermediate 126D. 2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)propanoic acid

[00414] To a homogeneous mixture of ethyl 2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)propanoate (92.7 mg, 0.3 mmol) in EtOH (3 mL), under nitrogen atmosphere, 2 M NaOH (aq) (0.3 mL, 0.6 mmol) was added dropwise. The resulting mixture was stirred at room temperature for 21 hours before being quenched with 4M HCl in dioxane (0.15 mL, 0.6 mmol). After stirring at room temperature for 30 minutes, the mixture was concentrated in vacuo to provide the product as a light brown solid, which was used without further purification, on a 100% basis, in the next step. MS(ES): m/z = 304 [M+H]+. tR = 0.39 minutes (Method A). Example 126. (±)-N-(4-Chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)propanamide

[00415] To the mixture of 2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)propanoic acid (30 mg, 0.1 mmol) and 4-chloroaniline (15.1 mg, 0.1 mmol) in anhydrous DMF (1.5 mL), under nitrogen atmosphere, DIPEA (0.06 mL, 0.3 mmol) was added followed by PyBOP (51.5 mg, 0.1 mmol). The resulting mixture was stirred at room temperature for 17.5 hours, before being diluted with DMF, filtered through a syringe filter, then purified by preparative HPLC/MS to provide the title compound as a racemate (11, 0 mg; 27% production). MS(ES): m/z = 413 [M+H]+. tR = 1.01 minutes (Method K). 1H NMR (500 MHz, DMSO-d6) δ 10.05 (s, 1H), 8.71 - 8.62 (m, 1H), 8.06 - 7.96 (m, 1H), 7.66 ( d, J=8.7 Hz, 2H), 7.64 - 7.57 (m, 2H), 7.35 (d, J=8.7 Hz, 2H), 7.04 (d, J=4 .9 Hz, 1H), 3.36 (q, J=6.8 Hz, 1H), 3.27 - 3.13 (m, 4H), 2.90 - 2.73 (m, 4H), 1 .25 (d, J=6.8 Hz, 3H). Example 127 (±)-N-(4-Bromophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)propanamide

[00416] Example 127 (17.1 mg; 37% production) was prepared following a procedure analogous to that for the synthesis of Example 126, except that 4-bromoaniline (20.4 mg, 0.12 mmol) was used instead of 4-chloroaniline. MS(ES): m/z = 457 [M+H]+. Tr = 1.04 minutes (Method K). 1H NMR (500 MHz, DMSO-d6) δ 10.58 (br. s., 1H), 8.65 (d, J=6.0 Hz, 1H), 8.09 - 7.98 (m, 1H ), 7.85 - 7.76 (m, 2H), 7.61 - 7.48 (m, 4H), 7.20 (d, J=6.1 Hz, 1H), 3.76 - 3, 66 (m, 2H), 3.39 - 3.27 (m, 2H), 3.27 - 3.16 (m, 2H), 2.95 - 2.82 (m, 1H), 2.58 - 2.54 (m, 2H), 1.46 (d, J=6.6 Hz, 3H). Example 128 (±)-2-(4-(6-Fluoroquinolin-4-yl)piperazin-1-yl)-N-phenylpropanamide

[00417] Example 128 (10.9 mg; 29% yield) was prepared following a procedure analogous to that for the synthesis of Example 126, except that aniline (11.1 mg, 0.12 mmol) was used instead of 4 - chloroaniline. MS(ES): m/z = 379 [M+H]+. Tr = 0.81 minutes (Method K). 1H NMR (500 MHz, DMSO-d6) δ 9.92 (s, 1H), 8.65 (d, J=4.9 Hz, 1H), 8.08 - 7.96 (m, 1H), 7 .67 - 7.54 (m, 4H), 7.30 (t, J=7.7 Hz, 2H), 7.12 - 7.00 (m, 2H), 3.36 (q, J=6 .7 Hz, 1H), 3.28 - 3.11 (m, 4H), 2.93 - 2.74 (m, 4H), 1.25 (d, J=6.8 Hz, 3H). Enantiomer 1 and Enantiomer 2 of racemic Example 126 Enantiomer 1: Example 129 N-(4-Chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)propanamide (absolute stereochemistry not determined homochiral) Enantiomer 2: Example 130 N-(4-Chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)propanamide (absolute stereochemistry not determined homochiral)

[00418] Racemic example 126 (11.0 mg) was purified by chiral SFC (70/30 CO2/MeOH mobile phase, Chiral column AD 25 x 3 cm, 5 μm, 85 ml/min, wavelength detector = 220 nm). Concentration of the appropriate fractions provided:

[00419] Example 129 (Enantiomer 1, first elution) (4.2 mg) MS(ES): m/z = 413 [M+H]+. Tr =1.04 minutes (Method K). 1H NMR (500 MHz, DMSO-d6): superimposed over racemate NMR.

[00420] Example 130 (Enantiomer 2, second elution) (4.1 mg) MS(ES): m/z = 413 [M+H]+. Tr =1.04 minutes (Method K). 1H NMR (500 MHz, DMSO-d6): superimposed over racemate NMR. Example 131 (±)-N-(4-Chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)butanamide

[00421] Example 131 (5.6 mg; 14% production) was prepared following a procedure analogous to that for the synthesis of Example 126, except that ethyl 2-bromobutanoate was used instead of ethyl 2-bromopropanoate. MS(ES): m/z = 427 [M+H]+. Tr = 1.09 minutes (Method K). 1H NMR (500 MHz, DMSO-d6) δ 10.17 (s, 1H), 8.65 (d, J=5.0 Hz, 1H), 8.01 (dd, J=9.0, 5, 6 Hz, 1H), 7.68 (d, J=8.8 Hz, 2H), 7.64 - 7.57 (m, 2H), 7.37 (d, J=8.8 Hz, 2H) , 7.03 (d, J=5.0 Hz, 1H), 3.58 - 3.54 (m, 1H), 3.25 - 3.11 (m, 4H), 2.95 - 2.76 (m, 4H), 1.85 - 1.60 (m, 2H), 0.89 (t, J=7.3 Hz, 3H). Example 132 (±)-2-(4-(6-Fluoroquinolin-4-yl)piperazin-1-yl)-N-phenylbutanamide

[00422] Example 132 (6.6 mg; 17% production) was prepared following a procedure analogous to that for the synthesis of Example 131, except that aniline was used instead of 4-chloroaniline. MS(ES): m/z = 393 [M+H]+. Tr = 0.91 minutes (Method K). 1H NMR (500 MHz, DMSO-d6) δ 10.08 (s, 1H), 8.61 (d, J=4.0 Hz, 1H), 7.98 (dd, J=8.8, 5, 6 Hz, 1H), 7.65 - 7.52 (m, 4H), 7.31 (t, J=7.8 Hz, 2H), 7.13 - 6.98 (m, 2H), 3, 24 - 3.09 (m, 5H), 2.96 - 2.76 (m, 4H), 1.83 - 1.60 (m, 2H), 0.87 (t, J=7.3 Hz, 3H). Example 133 (±)-N-(4-Chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)-3-methoxypropanamide

[00423] Example 133 (12.0 mg; 29% yield) was prepared following a procedure analogous to that for the synthesis of Example 126, except that methyl 2-bromo-3-methoxypropanoate was used instead of methyl 2-bromopropanoate. ethyl. MS(ES): m/z = 443 [M+H]+. Tr = 1.14 minutes (Method K). 1H NMR (500 MHz, DMSO-d6) δ 10.15 (s, 1H), 8.68 (d, J=4.4 Hz, 1H), 8.06 - 7.96 (m, 1H), 7 .71 (d, J=8.3 Hz, 2H), 7.65 - 7.56 (m, 2H), 7.38 (d, J=8.3 Hz, 2H), 7.04 (d, J=4.3 Hz, 1H), 3.83 - 3.64 (m, 2H), 3.62 - 3.36 (m, 1H), 3.18 (s, 3H), 3.01 - 2 .93 (m, 2H), 2.89 - 2.81 (m, 2H), 2.57 - 2.53 (m, 4H). Example 134 (±)-N-(4-Fluorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)-3-methoxypropanamide

[00424] Example 134 (11.3 mg; 35% production) was prepared following a procedure analogous to that for the synthesis of Example 133, except that 4-fluoroaniline was used instead of 4-chloroaniline. MS(ES): m/z = 427 [M+H]+. Tr = 1.05 minutes (Method K). 1H NMR (500 MHz, DMSO-d6) δ 10.07 (s, 1H), 8.67 (d, J=4.5 Hz, 1H), 8.09 - 7.98 (m, 1H), 7 .73 - 7.65 (m, 2H), 7.63 - 7.58 (m, 2H), 7.20 - 7.11 (m, 2H), 7.04 (d, J=4.5 Hz , 1H), 3.88 - 3.62 (m, 2H), 3.60 - 3.34 (m, 1H), 3.19 (s, 3H), 3.01 - 2.94 (m, 2H ), 2.89 - 2.82 (m, 2H), 2.56 - 2.52 (m, 4H). Examples 135 and 136 Enantiomer 1: N-(4-Chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)-3-methoxypropanamide (absolute stereochemistry not determined homo-chiral) Enantiomer 2: N-(4-Chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)-3-methoxypropanamide (absolute stereochemistry not determined homo-chiral)

[00425] Racemic example 133 (9.8 mg) was purified by chiral SFC (70/30 CO2/MeOH mobile phase, Chiral Lux-4 column 25 x 3 cm, 5 μm, 85 ml/min, length detector wavelength = 220 nm). Concentration of the appropriate fractions (previous elution) provided:

[00426] Example 135 (first elution) (3.2 mg) assigned as N-(4-chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)-3-methoxypropanamide ( Enantiomer 1). MS(ES): m/z = 443 [M+H]+. Tr =1.19 minutes (Method K). 1H NMR (500 MHz, DMSO-d6): superimposed over racemate NMR.

[00427] Example 136 (second elution) (3.3 mg) assigned as N-(4-chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)-3-methoxypropanamide ( Enantiomer 2). MS(ES): m/z = 443 [M+H]+. Tr =1.19 minutes (Method K). 1H NMR (500 MHz, DMSO-d6): superimposed over racemate NMR. Example 137 2-(4-((R)-3-((4-chlorophenyl)amino)-2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)-3-oxopropyl)phenyl acid )propanoic (mixture of two diastereomers) Intermediate 137A. tert-butyl 2-(4-(bromomethyl)phenyl)propanoate

[00428] To a solution of 2-(4-(bromomethyl)phenyl)propanoic acid (3 g, 12.34 mmol) in CH2Cl2 (100 mL) at RT was added oxalyl chloride (1.400 mL, 16.04 mmol) and 1 drop of DMF. The reaction was stirred at room temperature for 1 h. Then the mixture was concentrated to dryness. To the above reaction, CH2Cl2 (2 mL) was added followed by t-BuOH (100 mL). The mixture was stirred at room temperature for 16 h. The mixture was diluted with CH2Cl2 and washed with saturated NaHCO3, brine, dried over MgSO4, filtered and concentrated to provide Intermediate 137A (1.8 g, 6.02 mmol, 48.7% yield) as light yellow liquid. 1H NMR (400 MHz, chloroform-d) δ 7.36 (d, J=8.3 Hz, 2H), 7.33 - 7.23 (m, 2H), 4.60 (s, 1H), 4 .51 (s, 1H), 3.64 (dd, J=7.2, 2.8 Hz, 1H), 1.47 (dd, J=7.2, 1.2 Hz, 3H), 1, 42 (s, 9H) Intermediate 137B. 2-(4-((R)2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)-3-oxo-3-((R)-2-oxo-4-phenyloxazolidin-3 tert-butyl -yl)propyl)phenyl)propanoate (mixture of diastereomers)

[00429] To a solution of (R)-3-(2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)acetyl)-4-phenyloxazolidin-2-one (prepared from general procedures K , B, E, and L) (200 mg, 0.462 mmol) in THF (4 mL) at -40 °C was added NaHMDS (1 M in THF) (0.555 mL, 0.555 mmol) dropwise. The mixture was stirred at -40°C to -30°C for 20 min, then tert-butyl 2-(4-(bromomethyl)phenyl)propanoate (304 mg, 1.017 mmol) in THF (0.5 mL ) was added. The reaction was stirred at -20 °C for 16 h. The reaction was quenched with saturated NH4Cl and EtOAc at -20 °C. The organic was separated and washed with brine, dried over MgSO4, filtered and concentrated to dryness. This crude material was dissolved in DMF and purified with preparative HPLC (PHENOMENEX® Luna 5 μ 30 x 100 mm), flow rate of 40 mL/minutes with a gradient of 20% B-100% B for 10 minutes. Maintained at 100% B for 5 min. (A: 0.1% TFA in water/MeOH (90:10), B: 0.1% TFA in water/MeOH (10:90) monitoring at 254 nm. Combined fractions (tr=11.06 min ) containing the product. After concentration, 134 mg of Intermediate 137B (0.204 mmol, 44.1%) was obtained as a mixture of diastereomers. 1H NMR (400 MHz, chloroform-d) δ 9.15 (d, J=5, 1 Hz, 1H), 8.65 (dd, J=9.2, 4.9 Hz, 1H), 8.02 (d, J=3.5 Hz, 1H), 7.94 (dd, J= 9.3, 2.2 Hz, 1H), 7.89 - 7.73 (m, 1H), 7.36 - 7.24 (m, 4H), 7.13 (d, J=8.1 Hz , 1H), 7.17 (d, J=8.1 Hz, 1H), 6.99 (d, J=7.9 Hz, 2H), 6.84 - 6.64 (m, 2H), 5 .44 - 5.35 (m, 1H), 4.98 (br. s., 1H), 4.63 - 4.46 (m, 1H), 4.10 (ddd, J=8.9, 6 .5, 4.3 Hz, 1H), 3.69 - 3.60 (m, 1H), 3.52 (d, J=11.7 Hz, 1H), 3.03 (dt, J=13, 6, 4.2 Hz, 1H), 2.74 (ddd, J=13.6, 10.6, 6.8 Hz, 1H), 2.37 - 2.23 (m, 1H), 2.16 (d, J=12.8 Hz, 1H), 2.10 - 2.00 (m, 1H), 2.00 - 1.74 (m, 7H), 1.50 (dd, J=8.6 , 7.2 Hz, 3H), 1.45 - 1.29 (m, 9H) MS: Anal. Calculated for C40H43FN2O5 650.316, found [M+H] 651.3 LC: tr = 1.03 minutes (Method B ) Intermediate 137C. (2R)-3-(4-(1-tert-butoxy)-1-oxopropan-2-yl)phenyl)-2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)propanoic acid (mixture of diastereomers)

[00430] To a solution of Intermediate 137B (0.206 mmol, 134 mg) in THF (1.3 mL) at 0 °C, 30% H2O2 (0.093 mL, 0.824 mmol) was added dropwise, followed by the addition of LiOH at 2.7 M in H2O (0.122 mL, 0.329 mmol). The reaction was allowed to warm to room temperature and stirred at room temperature for 16 h. The reaction was carefully quenched at 0 °C by addition of saturated Na2SO3. The pH was adjusted to 5~6 with 1N HCl and the mixture was extracted with EtOAc. The combined organics were dried over MgSO4, filtered and concentrated. The crude material was purified with a 12 g, 30 mL/min ISCO column. 0-100% EtOAc/hexane in 35 min. The desired product was eluted with 22% EtOAc/hexane. After concentration, 35 mg (0.069 mmol, 33%) of 137C was obtained as a white solid. 1H NMR (400 MHz, chloroform-d) δ 8.82 (d, J=4.6 Hz, 1H), 8.15 (dd, J=9.2, 5.7 Hz, 1H), 7.68 (dd, J=10.5, 2.7 Hz, 1H), 7.49 (ddd, J=9.2, 7.9, 2.7 Hz, 1H), 7.41 (d, J=4 .5 Hz, 1H), 7.27 - 7.06 (m, 4H), 3.60 (d, J=7.0 Hz, 1H), 3.37 (br. s., 1H), 3, 04 - 2.96 (m, 2H), 2.95 - 2.81 (m, 1H), 2.12 (d, J=19.2 Hz, 1H), 2.00 - 1.76 (m, 8H), 1.45 (d, J=7.2 Hz, 3H), 1.42 - 1.34 (m, 9H) MS: Anal. Calculated for C31H36FNO4 505.263, found [M+H] 506.1 LC: tr = 0.92 minutes (Method B). Intermediate 137D. 2-(4-((R)-3-((4-chlorophenyl)amino)-2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)-3-oxopropyl)phenyl)propanoate tert-butyl (mixture of diastereomers)

[00431] To a solution of 137C (35 mg, 0.069 mmol) in CH2Cl2 (3 mL) at RT was added oxalyl chloride (8.90 μl, 0.104 mmol) dropwise followed by 1 drop of DMF. The reaction was stirred at room temperature for 2 h. Then the mixture was concentrated to dryness. To this mixture in THF (1 mL) at RT was added 4-chloroaniline (8.76 mg, 0.069 mmol), followed by Hunig's base (0.018 mL, 0.103 mmol). The reaction was stirred at room temperature for 3 h. The mixture was diluted with MeOH and purified with pre-parative HPLC (PHENOMENEX® Luna 5 μ 30 x 100 mm), flow rate of 40 mL/minutes with a gradient of 20% B-100% B for 10 minutes. Maintained at 100% B for 5min. (A: 0.1% TFA in water/MeOH (90:10), B: 0.1% TFA in water/MeOH (10:90) monitoring at 254 nm. Intermediate 137D (10 mg, 0.016 mmol, 46.4% production) was obtained as white solid. 1H NMR (400 MHz, chloroform-d) δ 9.06 (br. s., 1H), 8.57 (br. s., 1H), 7.95 - 7.84 (m, 2H), 7.84 - 7.63 (m, 1H), 7.26 - 7.10 (m, 8H), 3.66 - 3.49 (m, 4H) ), 3.06 (dd, J=13.6, 3.4 Hz, 2H), 2.87 (t, J=12.3 Hz, 1H), 2.70 (br. s., 1H), 2.40 (br. s., 1H), 2.15 (br. s., 1H), 2.10 - 1.80 (m, 7H), 1.41 (d, J=7.1 Hz, 3H), 1.37 - 1.30 (m, 9H) MS: Anal Calculated for C37H40ClFN2O3 614.271, found [M+H] 615.3 LC: tr = 1.06 minutes (Method B) Example 137. Acid 2 -(4-((R)-3-((4-chlorophenyl)amino)-2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)-3-oxopropyl)phenyl)propanoic acid

[00432] To Intermediate 137D (10 mg, 0.016 mmol) in a 2 dram vial was added 50% TFA/CH2Cl2 (0.3 mL, 0.016 mmol). The reaction was stirred at room temperature for 2 h. The reaction was concentrated to dryness and freeze-dried for 2 days. Example 137 (9.5 mg, 0.014 mmol, 83% yield) was obtained as a white solid. 1H NMR (400 MHz, chloroform-d) δ 8.74 - 8.52 (m, 1H), 8.34 (br. s., 1H), 7.90 - 7.78 (m, 2H), 7 .72 (t, J=7.9 Hz, 1H), 7.32 - 7.19 (m, 6H), 7.15 (d, J=8.7 Hz, 2H), 6.92 (d, J=8.6 Hz, 1H), 6.95 (d, J=8.3 Hz, 1H), 3.67 (d, J=6.7 Hz, 1H), 3.42 (br. s. , 1H), 3.13 - 2.83 (m, 3H), 2.30 (br. s., 1H), 2.15 (br. s., 1H), 2.02 (br. s., 1H), 1.97 - 1.87 (m, 3H), 1.84 (br. s., 3H), 1.41 (t, J=6.1 Hz, 3H) MS: Anal. Calculated for C33H32ClFN2O3 558.209, found [M+H] 559.3 LC: tr = 0.87 minutes (Method B) Enantiomer 1 and 2 of Racemic Example 137 Enantiomer 1: Example 138 acid 2-(4-((R)- 3-((4-chlorophenyl)amino)-2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)-3-oxopropyl)phenyl)propanoic (homochiral undetermined stereochemistry) Enantiomer 2: Example 139 2-(4-((R)-3-((4-chlorophenyl)amino)-2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)-3- acid oxopropyl)phenyl)propanoic (homochiral undetermined stereochemistry)

[00433] The diastereomers of Example 137 were purified by preparative SFC with the following conditions: Column: Chiral WHELK-O® KROMASIL® 25 x 3 cm ID, 5 μm particles; Mobile phase A: 70/30 CO2/MeOH; Detector wavelength: 220 nm; Flow: 85 mL/min. The appropriate fractions (Enantiomer 1 "Peak-1" tr = 15.2 minutes (Example 138) and Enantiomer 2 "Peak-2" tr = 17.2 minutes (Example 139) to give:

[00434] Example 138 (Enantiomer 1, first elution): 1H NMR (400 MHz, chloroform-d) δ 8.67 (br. s., 1H), 8.18 - 7.97 (m, 1H), 7 .67 (d, J=10.5 Hz, 1H), 7.47 (t, J=7.2 Hz, 1H), 7.33 - 7.23 (m, 6H), 7.17 (d, J=7.8 Hz, 2H), 7.08 (s, 4H), 7.02 (br. s., 1H), 3.76 (d, J=6.7 Hz, 1H), 3.36 (br. s., 1H), 3.03 (d, J=10.1 Hz, 1H), 2.87 (t, J=12.2 Hz, 1H), 2.63 (t, J=10 .0 Hz, 2H), 2.32 (br. s., 2H), 2.12 (br. s., 2H), 2.02 - 1.79 (m, 2H), 1.73 (d, J=10.0 Hz, 2H), 1.53 (d, J=7.1 Hz, 3H) MS: Anal. Calculated for C33H32ClFN2O3 558.209, found [M+H] 559.3 LC: tr = 0.86 minutes (Method B)

[00435] Example 139 (Enantiomer 2, first elution): 1H NMR (400 MHz, chloroform-d) δ 8.64 (br. s., 1H), 8.16 - 8.03 (m, 1H), 7 .66 (d, J=8.8 Hz, 1H), 7.45 (t, J=7.3 Hz, 1H), 7.30 (br. s., 7H), 7.20 - 7.01 (m, 7H), 3.74 (br. s., 1H), 3.35 (br. s., 1H), 3.00 (d, J=11.1 Hz, 1H), 2.96 - 2.80 (m, 1H), 2.71 - 2.55 (m, 1H), 2.31 (d, J=8.7 Hz, 1H), 2.25 - 2.13 (m, 1H) , 2.09 (br. s., 3H), 2.00 - 1.86 (m, 2H), 1.83 (br. s., 2H), 1.58 - 1.33 (m, 2H) , 1.30 - 1.25 (m, 2H), 1.00 - 0.71 (m, 1H) MS: Anal. Calculated for C33H32ClFN2O3 558.209, found [M+H] 559.3 LC: tr = 0.86 minutes (Method B) Example 140 2-(4-(( R )-3-((4-cyanophenyl)amino)- acid 2-((cis)-4-(6-fluoroquinolin-4-yl)cyclohexyl)-3-oxopropyl)phenyl)propanoic acid

[00436] Example 140 was obtained following the procedures in Example 137 using the corresponding 4-cyanoaniline. 1H NMR (400 MHz, chloroform-d) δ 8.99 (br. s., 1H), 8.42 (br. s., 1H), 8.06 (br. s., 1H), 7.95 (d, J=9.3 Hz, 1H), 7.83 (t, J=8.2 Hz, 1H), 7.50 - 7.35 (m, 3H), 7.25 - 7.10 ( m, 4H), 3.78 - 3.54 (m, 1H), 3.53 (s, 1H), 3.18 - 2.92 (m, 2H), 2.92 - 2.65 (m, 1H), 2.38 (br. s., 1H), 2.27 - 2.16 (m, 1H), 2.03 (s, 2H), 2.00 - 1.93 (m, 3H), 1.91 (br. s., 3H), 1.80 (br. s., 1H), 1.52 - 1.36 (m, 3H), 0.94 - 0.69 (m, 1H) MS : Anal. Calculated for C34H32FN3O3 549.243, found [M+H] 550.3 LC: tr = 0.83 minutes (Method B). Examples 141 and 142

[00437] These compounds were obtained following the procedures in Example 137 using the corresponding carboxylic acids (which can be prepared using General Procedures K, B, E or 58A) and cyclohexylamine. Table 6 Example 143 (±)-N-(4-chlorophenyl)-2-( cis- and trans-4-(pyrazolo[1,5-a]pyrimidin-7-yl)cyclohexyl)propanamide (mixture of four isomers) Intermediate 143A. Ethyl 2-(1,4-dioxaspiro[4,5]decan-8-ylidene)propanoate

[00438] To a suspension of NaH (0.307 g, 7.68 mmol) in THF (8 mL) cooled to 0 °C was added ethyl 2-(diethoxyphosphoryl)propanoate (1.830 g, 7.68 mmol) slowly. After 30 min, 1,4-dioxaspiro[4.5]decan-8-one (1 g, 6.40 mmol) was added. The resulting mixture was stirred at 0°C for 2 hours, then warmed to room temperature overnight. The mixture was quenched with water, THF was removed under reduced pressure. The residue was dissolved in EtOAc, washed with water, brine, dried over Na2SO4 and concentrated. The crude was purified by ISCO (EtOAc/Hex 0-30%). The product-containing fractions were concentrated to yield Intermediate 143A (1.2 g, 78% production) as a light yellow oil. 1H NMR (400 MHz, CHLOROFORM-d) β 4.19 (q, J=7.1 Hz, 2H), 4.03 - 3.89 (m, 4H), 2.68 - 2.53 (m, 2H), 2.46 - 2.28 (m, 2H), 1.89 (s, 3H), 1.78 - 1.66 (m, 4H), 1.30 (t, J=7.1 Hz , 3H) Intermediate 143B. Ethyl 2-(1,4-dioxaspiro[4,5]decan-8-yl)propanoate

[00439] A suspension of Intermediate 143A (500 mg, 2.081 mmol)(1A) and 10% palladium on carbon (25 mg, 0.024 mmol) in EtOAc (5 mL) was hydrogenated on a Parr shaker at 3.16 kg/ cm2 (45 psi) for 6 h. The catalyst was filtered, and the filtrate was concentrated to yield Intermediate 143B (450 mg, 89% yield) as a clear oil. 1H NMR (400 MHz, CHLOROFORM-d) β 4.12 (dtt, J=10.7, 7.1, 3.6 Hz, 2H), 3.98 - 3.81 (m, 4H), 2, 35 - 2.17 (m, 1H), 1.83 - 1.68 (m, 3H), 1.66 - 1.45 (m, 4H), 1.43 - 1.28 (m, 2H), 1.27 - 1.22 (m, 3H), 1.14 - 1.07 (m, 3H) Intermediate 143C. Ethyl 2-(4-oxocyclohexyl)propanoate

[00440] To a solution of ethyl 2-(1,4-dioxaspiro[4,5]decan-8-yl)propanoate (450 mg, 1.857 mmol)(1B) in THF (5 mL) was added hydrogen chloride at 1 M (aqueous) (0.929 mL, 3.71 mmol). The mixture was heated to 50°C for 6 h. The reaction mixture was concentrated. The residue was dissolved in EtOAc, washed with water (2X), brine, dried over Na2SO4 and concentrated. The crude was purified with ISCO (EtOAc/Hex 0-30%). The product-containing fractions were concentrated to yield Intermediate 143C (290 mg, 79% production) as a clear oil. 1H NMR (400 MHz, CHLORO-FORMIUM-d) β 4.22 - 4.06 (m, 2H), 2.46 - 2.30 (m, 5H), 2.13 - 1.91 (m, 3H ), 1.56 - 1.42 (m, 2H), 1.31 - 1.24 (m, 3H), 1.18 (d, J=7.1 Hz, 3H) Intermediate 143D. Ethyl 2-(4-(((trifluoromethyl)sulfonyl)oxy)cyclohex-3-en-1-yl)propanoate

[00441] Intermediate 143C (200 mg, 1.01 mmol)(1C) and 2,6-di-tert-butyl-4-methylpyridine (238 mg, 1.16 mmol) were dissolved in dry DCM (10 ml). To the reaction mixture, trifluoromethanesulfonic anhydride (0.186 mL, 1.11 mmol) was added dropwise and stirred for 2 h. The suspension was filtered and the filtrate was diluted with DCM, washed with 1N HCl (2X), saturated sodium bicarbonate solution, water, brine and dried over Na2SO4 and concentrated to yield Intermediate 143D (320 mg, 96% yield ) as a brown oil. 1H NMR (400 MHz, CHLOROFORM-d) β 5.73 (t, J=6.1 Hz, 1H), 4.28 - 4.05 (m, 2H), 2.52 - 2.17 (m, 4H), 2.08 - 1.79 (m, 3H), 1.49 (dt, J=11.1, 6.6 Hz, 1H), 1.31 - 1.20 (m, 3H), 1 .19 - 1.04 (m, 3H) Intermediate 143E. Ethyl 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)propanoate

[00442] To a solution of Intermediate 143D (300 mg, 0.908 mmol)(1D) in DMSO (5 mL) was added 4,4,4',4',5,5,5',5'-octamethyl-2 ,2'-bi(1,3,2-dioxaborolane) (230 mg, 0.908 mmol) and potassium acetate (267 mg, 2.72 mmol). After the mixture was degassed with N2 for 10 min, PdCl2(dppf) (19.9 mg, 0.027 mmol) was added. The mixture was heated to 80°C overnight. The mixture was partitioned between EtOAc and water. The organic phase was concentrated and purified by ISCO. The product-containing fractions were concentrated to produce Intermediate 143E (168 mg, 60% production) as a brown oil. 1H NMR (400 MHz, CHLOROFORM-d) β 6.66 - 6.40 (m, 1H), 4.31 - 4.00 (m, 2H), 2.34 - 2.26 (m, 1H), 2.25 - 2.19 (m, 1H), 2.19 - 2.04 (m, 2H), 1.95 - 1.75 (m, 3H), 1.73 - 1.60 (m, 1H ), 1.29 - 1.24 (m, 15H), 1.13 (dd, J=11.6, 7.0 Hz, 3H) Intermediate 143F. Ethyl 2-(4-(pyrazolo[1,5-a]pyrimidin-7-yl)cyclohex-3-en-1-yl)propanoate

[00443] A mixture of 7-chloropyrazolo[1,5-a]pyrimidine (0.193 g, 1.260 mmol), Intermediate 143E (0.400 g, 1.298 mmol), Na2CO3 (0.534 g, 5.04 mmol), and Pd(Ph3P )4 (0.073 g, 0.063 mmol) in dioxane (11.67 ml) and water (3.89 ml) was heated at 100 °C overnight. The reaction was quenched with water and diluted with EtOAc. The layers were separated. The aqueous phase was extracted with EtOAc (3X). The organics were combined, dried over Na2SO4, filtered, and concentrated to provide a brown residue. Purification of the crude material by silica gel chromatography using an ISCO machine (40 g column, 40 mL/min, 0-70% EtOAc in hexanes for 16 min, tr = 10.5 min) provided 143F (0.224 g, 0.748 mmol, 59.4% production) as a yellow residue. ESI MS (M+H)+ = 300.2. HPLC peak tr = 0.95 minutes. HPLC conditions: method A.Intermediate 143G.2-(4-(pyrazolo[1,5-a]pyrimidin-7-yl)cyclohexyl)ethyl propanoate

[00444] To a solution of 143F (0.224 g, 0.748 mmol) in MeOH (3.74 ml) was added ammonium formate (0.236 g, 3.74 mmol) followed by Pd/C (0.021 g, 0.202 mmol). The reaction was heated at 70 °C for 1 h. The reaction was filtered through CELITE® and the filter mass washed with CH2Cl2. The filtrate was concentrated. The raw material was taken up in EtOAc and washed with a saturated aqueous solution of NaHCO3 (1X). The organic phase was dried over Na2SO4, filtered, and concentrated to provide 143G (220 mg, 98%) as a yellow residue. ESI MS (M+H)+ = 302.2. HPLC peak tr = 0.94 minutes. HPLC conditions: method A.Example 143. (±)-N-(4-chlorophenyl)-2-( cis- and trans -4-(pyrazolo[1,5- a]pyrimidin-7-yl)cyclohexyl)propanamide (mixture of four isomers)

[00445] To a solution of 4-chloroaniline (92 mg, 0.720 mmol) in THF (0.9 mL) at 0 °C was added a solution of isopropymagnesium chloride (360 μl, 0.720 mmol). The resulting solution was warmed to room temperature and stirred for 5 min, then 143G (108.5 mg, 0.360 mmol) in THF (0.9 mL) was added dropwise. The reaction was heated to 70 °C for 2.5 h, then allowed to cool to room temperature. More 4-chloroaniline (42 mg) and isopropylmagnesium chloride (360 μl, 0.720 mmol) were added. The reaction was quenched with a saturated aqueous solution of NH4Cl and diluted with EtOAc. The layers were separated. The aqueous phase was extracted with EtOAc (3X). The combined organic phases were dried over Na2SO4, filtered, and concentrated to provide a residue. The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 40-80% B for 20 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to provide the title compound as a mixture of 4 isomers (43.4 mg, 31%). ESI MS (M+H)+ = 383.1. HPLC peak tr = 0.96 minutes. Purity = 99%. HPLC conditions: method A.Examples 144 (a), (b), (c), and (d) (S )-N-(4-chlorophenyl)-2-(( cis )-4-(pyrazolo[1 ,5-a]pyrimidin-7-yl)cyclohexyl)propanamide and (R)-N-(4-chlorophenyl)-2-((cis)-4-(pyrazolo[1,5-a]pyrimidin-7-yl )cyclohexyl)propanamide and (S)-N-(4-chlorophenyl)-2-((trans)-4-(pyrazolo[1,5-a]pyrimidin-7-yl)cyclohexyl)propanamide and (R)-N -(4-chlorophenyl)-2-((trans)-4-(pyrazolo[1,5-a]pyrimidin-7-yl)cyclohexyl)propanamide (homochiral relative and absolute stereochemistry was undetermined and arbitrarily specified)

[00446] Approximately 43.4 mg of diastereomeric and racemic Example 143 were resolved. The isomeric mixture was purified by preparative SFC with the following conditions: Column: Chiral IE, 25 x 3 cm ID, 5 μm particles; Mobile phase A: 80/20 CO2/MeOH; Detector wavelength: 220 nm; Flow: 85 mL/min. The fractions (144(a) "Peak-1" tr = 13.272, 144(b) "Peak-2" tr = 14.097, 144(c) "Peak-3" tr = 19.986, 144(d) "Peak-4 " tr = 27.958; analytical conditions: Column: Chiral IE, 250 x 4.6 mm ID, 5 μm particles; Mobile phase A: 80/20 CO2/MeOH; Flow: 2.0 mL/min) were collected in MeOH. The stereoisomeric purity of peak 2 and peak 3 was estimated to be greater than 99% based on preparative SFC chromatograms. Peak 1 (23.8 mg) was purified again by preparative SFC with the following conditions: Column: Chiral OJ, 25 x 3 cm ID, 5 μm particles; Mobile phase A: 80/20 CO2/MeOH; Detector wavelength: 220 nm; Flow: 85 mL/min. The fractions ("Peak-1" tr = 4.558 and "Peak-2" tr = 5.622; analytical conditions: Column: Chiral OJ, 250 x 4.6 mm ID, 5 μm particles; Mobile phase A: 80/20 of CO2/MeOH; Flow: 2.0 mL/min) were collected in MeOH. The stereoisomeric purity of the fractions was estimated to be greater than 99% based on the preparative SFC chromatogram. Each diastereomer or enantiomer was also purified using preparative LC/MS:

[00447] Example 144(a) First elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: Waters XBridge c-18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 40-100% B for 20 minutes, then a 10-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 1 (11.3 mg, 8.2%). ESI MS (M+H)+ = 383.2. HPLC peak tr = 1.833 minutes. Purity = 100%. HPLC conditions: B. Absolute stereochemistry not determined.

[00448] Example 144(b) Second elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: Waters XBridge c-18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 40-100% B for 20 minutes, then a 10-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 2 (11.1 mg, 8.1%). ESI MS (M+H)+ = 382.9. HPLC peak tr = 1.829 minutes. Purity = 100%. HPLC conditions: B. Absolute stereochemistry not determined.

[00449] Example 144(c) Third elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: Waters XBridge c-18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 40-100% B for 20 minutes, then a 10-minute hold at 100% B; Flow: 20 mL/min. Fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 3 (7.2 mg, 5.0%). ESI MS (M+H)+ = 383.2. HPLC peak tr = 1.874 minutes. Purity = 96%. HPLC conditions: B. Absolute stereochemistry not determined.

[00450] Example 144(d) Fourth elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: Waters XBridge c-18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 40-100% B for 20 minutes, then a 10-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 4 (7.6 mg, 5.1%). ESI MS (M+H)+ = 383.3. HPLC peak tr = 1.874 minutes. Purity = 93%. HPLC conditions: B. Absolute stereochemistry not determined. Example 147 (±)-2-( Cis-and trans -4-(1,8-naphthyridin-4-yl)cyclohexyl)-N-(4-chlorophenyl)propanamide (mixture of four isomers) Intermediate 147A. Ethyl 2-(4-(1,8-naphthyridin-4-yl)cyclohex-3-en-1-yl)propanoate

[00451] A mixture of 4-bromo-1,8-naphthyridine (0.070 g, 0.335 mmol), Intermediate 143E (0.106 g, 0.345 mmol), Na2CO3 (0.142 g, 1.339 mmol), and Pd(Ph3P)4 (0.019 g, 0.017 mmol) in dioxane (3.10 ml) and water (1.034 ml) was heated at 100 °C overnight. The reaction was quenched with water and diluted with EtOAc. The layers were separated. The aqueous phase was extracted with EtOAc (2X). The organics were combined, dried over Na2SO4, filtered, and concentrated to provide a brown residue. Purification of the crude material by silica gel chromatography using an ISCO machine (24 g column, 35 mL/min, 0-20% MeOH in CH2Cl2 for 25 minutes, tr = 17 min) gave 147A (92.7 mg, 0.299 mmol, 89% yield) as a yellow residue. ESI MS (M+H)+ = 311.2. HPLC peak tr = 0.72 minutes. HPLC conditions: method A.Intermediate 147B. Ethyl 2-(4-(1,8-naphthyridin-4-yl)cyclohexyl)propanoate

[00452] To a solution of 147A (0.0927 g, 0.299 mmol) in MeOH (1.493 ml) was added ammonium formate (0.094 g, 1.493 mmol) followed by 10% Pd/C (8.58 mg, 0.081 mmol). The reaction was heated at 70 °C for 1 h. The reaction was filtered through CELITE® and the filter mass washed with CH2Cl2. The filtrate was concentrated. The raw material was taken up in EtOAc and washed with a saturated aqueous solution of NaHCO3 (1X). The organic phase was dried over Na2SO4, filtered, and concentrated to provide 147B (72.5 mg, 78%) as a brown residue. ESI MS (M+H)+ = 313.3. HPLC peak tr = 0.70 minutes. HPLC conditions: method A.Example 147. (±)-2-(Cis- and trans-4-(1,8-naphthyridin-4-yl)cyclohexyl)-N-(4-chlorophenyl)propanamide (mixture of four isomers)

[00453] To a solution of 4-chloroaniline (0.059 g, 0.464 mmol) in THF (0.4 mL) at 0 °C was added a solution of isopropylmagnesium chloride (0.232 mL, 0.464 mmol). The resulting solution was warmed to room temperature and stirred for 5 min, then 147B (0.0725 g, 0.232 mmol) in THF (0.76 mL) was added dropwise. The reaction was heated at 70 °C for 3 h, then allowed to cool to room temperature. The reaction was quenched with a saturated aqueous solution of NH4Cl and diluted with EtOAc. The layers were separated. The aqueous phase was extracted with EtOAc (3X). The combined organic phases were dried over Na2SO4, filtered, and concentrated to provide a residue. The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 30 to 70% B for 19 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to provide the title compound as a mixture of 4 isomers (33.4 mg, 36%). ESI MS (M+H)+ = 394.2. HPLC peak tr = 1.743 minutes. Purity = 98%. HPLC conditions: method B. Example 148 (a), (b), (c), and (d) (S )-2-((cis)-4-(1,8-Naphthyridin-4-yl)cyclohexyl )-N-(4-chlorophenyl)propanamide and (R )-2-(( cis )-4-(1,8-naphthyridin-4-yl)cyclohexyl)-N-(4-chlorophenyl)propanamide and (S ) -2-(( trans )-4-(1,8-naphthyridin-4-yl)cyclohexyl)-N-(4-chlorophenyl)propanamide and (R )-2-(( trans )-4-(1,8 -naphthyridin-4-yl)cyclohexyl)-N-(4-chlorophenyl)propanamide (homochiral relative and absolute stereochemistry not determined and arbitrarily specified)

[00454] Approximately 34.3 mg of diastereomeric and racemic Example 147 were resolved. The isomeric mixture was purified by preparative SFC with the following conditions: Column: Chiral AD, 25 x 3 cm ID, 5 μm particles; Mobile phase A: 70/30 CO2/MeOH; Detector wavelength: 220 nm; Flow: 85 mL/min. The fractions ("Peak-1" tr = 7.377, "Peak-2" tr = 8.774, "Peak-3" tr = 10.106, "Peak-4" tr = 14.282; analytical conditions: Column: Chiral AD, 250 x 4 .6 mm ID, 5 μm particles; Mobile phase A: 70/30 CO2/MeOH; Flow: 2.0 mL/min) were collected in MeOH. The stereoisomeric purity of each fraction was estimated to be greater than 99% based on preparative SFC chromatograms. Each diastereomer or enantiomer was also purified using preparative LC/MS:

[00455] Example 148 (a) First elution isomer: The crude material was purified by means of preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 30 to 70% B for 19 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 1 (5.3 mg, 5.7%). ESI MS (M+H)+ = 394.1. HPLC peak tr = 1.757 minutes. Purity = 99%. HPLC conditions: method B. Absolute stereochemistry not determined.

[00456] Example 148 (b) Second elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 30 to 70% B for 19 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 2 (6.0 mg, 6.4%). ESI MS (M+H)+ = 394.1. HPLC peak tr = 1.719 minutes. Purity = 98%. HPLC conditions: method B. Absolute stereochemistry not determined.

[00457] Example 148 (c) Third elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 30 to 70% B for 19 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 3 (6.1 mg, 6.3%). ESI MS (M+H)+ = 394.2. HPLC peak tr = 1.694 minutes. Purity = 95%. HPLC conditions: method B. Absolute stereochemistry not determined.

[00458] Example 148 (d) Fourth elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 30 to 70% B for 19 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 4 (3.5 mg, 3.8%). ESI MS (M+H)+ = 394.3. HPLC peak tr = 1.743 minutes. Purity = 99%. HPLC conditions: method B. Absolute stereochemistry not determined. Example 149 (±)-2-( Cis- and trans -4-(1H-pyrazolo[4,3-b]pyridin-7-yl)cyclohexyl)-N-(4-chlorophenyl)propanamide (mixture of four isomers) Intermediate 149A. Ethyl 2-(4-( 1 H-pyrazolo[4,3-b]pyridin-7-yl)cyclohex-3-en-1-yl)propanoate

[00459] A mixture of 7-chloro-1H-pyrazolo[4,3-b]pyridine (0.048 g, 0.315 mmol), Intermediate 143E (0.100 g, 0.324 mmol), Na2CO3 (0.134 g, 1.260 mmol), and 10 % Pd(Ph3P)4 (0.018 g, 0.016 mmol) in dioxane (2.92 ml) and water (0.972 ml) was heated at 100 °C overnight. The reaction was quenched with water and diluted with EtOAc. The layers were separated. The aqueous phase was extracted with EtOAc (3X). The organics were combined, dried over Na2SO4, filtered, and concentrated to provide a brown residue. Purification of the crude material by silica gel chromatography using an ISCO machine (40 g column, 40 mL/min, 0-100% EtOAc in hexanes for 23 min, tr = 18 min) provided 149A (0.039 g, 0.124 mmol , 39.3% of production) as a yellow residue. ESI MS (M+H)+ = 300.2. HPLC peak tr = 0.69 minutes. HPLC conditions: method A. Intermediate 149B.2-(4-( 1 H-pyrazolo[4,3-b]pyridin-7-yl)cyclohexyl)ethyl propanoate

[00460] To a solution of 149A (0.0395 g, 0.132 mmol) in MeOH (0.660 ml) was added ammonium formate (0.042 g, 0.660 mmol) followed by Pd/C (3.79 mg, 0.036 mmol). The reaction was heated at 70 °C for 1 h. More ammonium formate (0.042 g, 0.660 mmol) was added and the reaction was heated for 1 hour at 70 °C, then allowed to cool to room temperature. The reaction was filtered through CELITE® and the filter mass washed with CH2Cl2. The filtrate was concentrated. The raw material was taken up in EtOAc and washed with a saturated aqueous solution of NaHCO3 (1X). The organic phase was dried over Na2SO4, filtered, and concentrated to provide 149B (34.3 mg, 86%) as a yellow residue. It will go ahead raw. ESI MS (M+H)+ = 302.4. HPLC peak tr = 0.66 minutes. HPLC conditions: method A.Example 149. (±)-2-( Cis- and trans -4-(1H-pyrazolo[4,3-b]pyridin-7-yl)cyclohexyl)-N-(4-chlorophenyl )propanamide (mixture of four isomers)

[00461] To a solution of 4-chloroaniline (58.1 mg, 0.455 mmol) in THF (0.3 mL) at 0 °C was added a solution of isopropymagnesium chloride (228 μl, 0.455 mmol). The resulting solution was warmed to room temperature and stirred for 5 min, then 149B (34.3 mg, 0.114 mmol) in THF (0.3 mL) was added dropwise. The reaction was heated to 70 °C for 2 h, then allowed to cool to room temperature. The reaction was quenched with a saturated aqueous solution of NH4Cl and diluted with EtOAc. The layers were separated. The aqueous phase was extracted with EtOAc (3X). The combined organic phases were dried over Na2SO4, filtered, and concentrated to provide a residue. The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 20-60% B for 19 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to provide the title compound as a mixture of 4 isomers (25.4 mg, 58%). ESI MS (M+H)+ = 383.3. HPLC peak tr = 1.662 minutes. Purity = 99%. HPLC conditions: method B. Example 150 (a), (b), (c), and (d) (S )-2-((cis)-4-(1H-Pyrazolo[4,3-b]pyridine -7-yl)cyclohexyl)-N-(4-chlorophenyl)propanamide and (R )-2-(( cis )-4-(1H-pyrazolo[4,3-b]pyridin-7-yl)cyclohexyl) -N-(4-chlorophenyl)propanamide and (S)-2-((trans)-4-(1H-pyrazolo[4,3-b]pyridin-7-yl)cyclohexyl)-N-(4-chlorophenyl )propanamide and (R )-2-(( trans )-4-(1 H-pyrazolo[4,3-b]pyridin-7-yl)cyclohexyl)-N-(4-chlorophenyl)propanamide (relative and absolute stereochemistry undetermined homochiral and arbitrarily specified ar)

[00462] Approximately 29.3 mg of diastereomeric and racemic Example 149 were resolved. The isomeric mixture was purified using preparative SFC with the following conditions: Column: Chiral WHELK-O®, 25 x 3 cm ID, 5 μm particles; Mobile phase A: 70/30 CO2/MeOH; Detector wavelength: 220 nm; Flow: 85 mL/min. The fractions (150(a) "Peak-1" tr = 9.587, 150(b) "Peak-2" tr = 10.407, 150(c) "Peak-3" tr = 11.794, 15-(d) "Peak- 4" tr = 12.855; analytical conditions: Column: Chiral WHELK-O®, 250 x 4.6 mm ID, 5 μm particles; Mobile phase A: 80/20 CO2/MeOH; Flow: 2.0 mL/min ) were collected in MeOH. The stereoisomeric purity of peaks 1, 3, and 4 was estimated to be greater than 95% based on preparative SFC chromatograms. Peak 2 was purified again by preparative SFC with the following conditions: Column: Chiral AS, 25 x 3 cm ID, 5 μm particles; Mobile phase A: 80/20 CO2/MeOH; Detector wavelength: 220 nm; Flow: 85 mL/min. The fractions ("Peak-1" tr = 3.391 and "Peak-2" tr = 4.071; analytical conditions: Column: Chiral AS, 250 x 4.6 mm ID, 5 μm particles; Mobile phase A: 80/20 of CO2/MeOH; Flow: 2.0 mL/min) were collected in MeOH. The stereoisomeric purity of the fractions was estimated to be greater than 99% based on the preparative SFC chromatogram. Each diastereomer or enantiomer was also purified using preparative LC/MS:

[00463] Example 150(a) First elution isomer: The crude material was purified by means of preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 20-60% B for 19 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 1 (4.6 mg, 11%). ESI MS (M+H)+ = 383.1. HPLC peak tr = 1.611 minutes. Purity = 100%. HPLC conditions: method B. Absolute stereochemistry not determined.

[00464] Example 150(b) Second elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 20-60% B for 19 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 2 (4.6 mg, 11%). ESI MS (M+H)+ = 383.2. HPLC peak tr = 1,630 minutes. Purity = 100%. HPLC conditions: method B. Absolute stereochemistry not determined.

[00465] Example 150(c) Third elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 20-60% B for 19 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 3 (9.1 mg, 21%). ESI MS (M+H)+ = 383.2. HPLC peak tr = 1.659 minutes. Purity = 100%. HPLC conditions: method B. Absolute stereochemistry not determined.

[00466] Example 150(d) Fourth elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 20-60% B for 19 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 4 (4.7 mg, 10%). ESI MS (M+H)+ = 383.3. HPLC peak tr = 1.704 minutes. Purity = 93%. HPLC conditions: method B. Absolute stereochemistry not determined.Example 151 (±)-N-(Cis-e trans -4-chlorophenyl)-2-(4-(6-iodoquinolin-4-yl)cyclohexyl)propanamide Intermediate 151A.2-(4-(6-nitroquinolin-4-yl)cycloex-3-en-1-yl)ethyl propanoate

[00467] A 350 mL sealed tube was charged with a mixture of 4-chloro-6-nitroquinoline (2 g, 9.59 mmol), Intermediate 143E (3.04 g, 9.88 mmol), Na2CO3 (4. 06 g, 38.4 mmol), and Pd(Ph3P)4 (0.554 g, 0.479 mmol) in dioxane (89 mL) and water (29.6 mL). The reaction was heated to 100 °C overnight. The reaction was quenched with water and diluted with EtOAc. The layers were separated. The aqueous phase was extracted with EtOAc (3X). The organics were combined, dried over Na2SO4, filtered, and concentrated to provide a brown residue. Purification of the crude material by silica gel chromatography using an ISCO machine (80 g column, 60 mL/min, 0-45% EtOAc in hexanes for 19 min, tr = 14 min) provided 151A (2.955 g, 8.34 mmol, 87% production) as a yellow residue. ESI MS (M+H)+ = 355.2. HPLC peak tr = 0.98 minutes. HPLC conditions: method A.Intermediate 151B. Ethyl 2-(4-(6-aminoquinolin-4-yl)cyclohexyl)propanoate

[00468] To a solution of 151A (0.455 g, 1.284 mmol) in MeOH (6.42 ml) was added ammonium formate (0.405 g, 6.42 mmol) followed by 10% Pd/C (0.037 g, 0.347 mmol). The reaction was heated at 70 °C for 1 h. The reaction was filtered through CELITE® and the filter mass washed with CH2Cl2. The filtrate was concentrated. The raw material was taken up in EtOAc and washed with a saturated aqueous solution of NaHCO3 (2X). The organic phase was dried over Na2SO4, filtered, and concentrated to provide 151B (379 mg, 90%) as a brown residue. NMR showed pure desired product at 1.8:1 dr. ESI MS (M+H)+ = 327.3. HPLC peak tr = 0.71 minutes. HPLC conditions: method A.Intermediate 151C. Ethyl 2-(4-(6-iodoquinolin-4-yl)cyclohexyl)propanoate

[00469] A solution of 151B (0.379 g, 1.161 mmol) and aq. HCl. (0.59 mL) in water (2.1 mL) was cooled to 0 °C, then a solution of sodium nitrite (0.096 g, 1.393 mmol) in water (2.1 mL) was added. A solution of potassium iodide (0.289 g, 1.742 mmol) in water (2.1 mL) was added dropwise to the above solution after the solid dissolved completely. After addition, the mixture was stirred for 30 minutes at rt, then heated at 70°C for 1 h. After cooling, the solution was neutralized by slowly adding a Na2S2O3 solution (1.81 mL), then extracted with CH2Cl2 (2X). The organic phase was washed with water, dried over Na2SO4, filtered, and concentrated to provide a brown residue. The raw material was dissolved in a minimum amount of CH2Cl2 and subjected to chromatography. Purification of the crude material by silica gel chromatography using an ISCO machine (40 g column, 40 mL/min, 0-55% EtOAc in hexanes for 15 minutes, tr = 10.5 min) gave 151C (92.7 mg, 0.212 mmol, 18.26% yield) as a yellow residue. ESI MS (M+H)+ = 438.1. HPLC peak tr = 0.89 minutes. HPLC conditions: method A. Example 151. (±)-N-(Cis- and trans-4-chlorophenyl)-2-(4-(6-iodoquinolin-4-yl)cyclohexyl)propanamide

[00470] To a solution of 4-chloroaniline (0.464 g, 3.64 mmol) in THF (2.8 mL) at 0 °C was added a solution of isopropylmagnesium chloride (1.820 mL, 3.64 mmol) . The resulting solution was warmed to room temperature and stirred for 5 min, then 151C (0.796 g, 1.820 mmol) in THF (4.8 mL) was added dropwise. The reaction was heated to 70 °C for 2 h, then allowed to cool to room temperature. More isopropylmagnesium chloride (1.820 mL, 3.64 mmol) was added. The reaction was heated for another 2 h. The reaction was quenched with a saturated aqueous solution of NH4Cl and diluted with EtOAc. The layers were separated. The aqueous phase was extracted with EtOAc (3X). The combined organic phases were dried over Na2SO4, filtered, and concentrated to provide a residue. Purification of the crude material by silica gel chromatography using an ISCO machine (80 g column, 60 mL/min, 0-65% EtOAc in hexanes for 35 min, tr = 27 min) provided (±)-trans-Example 151 (455 mg, 0.702 mmol, 39% production) and (±)-cis-Example 151 (111 mg, 12%). The trans diastereomer elutes first followed by the cis diastereomer. ESI MS (M+H)+ = 519.1. Example 152 (a), (b), (c), and (d) (S )-N-(4-Chlorophenyl)-2-((cis)-4-(6-iodoquinolin-4-yl)cyclohexyl) propanamide and (R)-N-(4-chlorophenyl)-2-((cis)-4-(6-iodoquinolin-4-yl)cyclohexyl)propanamide and (S)-N-(4-chlorophenyl)-2- ((trans)-4-(6-iodoquinolin-4-yl)cyclohexyl)propanamide and (R)-N-(4-chlorophenyl)-2-((trans)-4-(6-iodoquinolin-4-yl) cyclohexyl)propanamide (homochiral relative and absolute stereochemistry not determined and arbitrarily specified)

[00471] Approximately 65.1 mg of diastereomeric and racemic Example 9 were resolved. The isomeric mixture was purified by preparative SFC with the following conditions: Column: OJ-H, 25 x 3 cm ID, 5 μm particles; Mobile phase A: 80/20 CO2/MeOH; Detector wavelength: 220 nm; Flow: 150 mL/min. The fractions (152(a) "Peak-1" tr = 4.64 min, 152(b) "Peak-2" tr = 5.35 min, 152(c) and 152(d) "Peak-3" tr = 6.43 min) were collected in MeOH. The stereoisomeric purity of peaks 1 and 2 was estimated to be greater than 95% based on preparative SFC chromatograms. Peak 3 was again purified by preparative SFC with the following conditions to provide Isomers 3 and 4: Column: Lux-Cellulose, 25 x 3 cm ID, 5 μm particles; Mobile phase A: 75/25 CO2/MeOH; Detector wavelength: 220 nm; Flow: 180 mL/min. The fractions (152(a) "Peak-1" tr = 7.63 minutes and 152(b) "Peak-2" tr = 8.6 min) were collected in MeOH. The stereoisomeric purity of the fractions was estimated to be greater than 95% based on preparative SFC chromatograms. Each diastereomer or enantiomer was also purified using preparative LC/MS:

[00472] Example 152(a) First elution isomer: The crude material was purified by means of preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 50-100% B for 20 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 1 (14.5 mg, 12%). ESI MS (M+H)+ = 519.2. HPLC peak tr = 2.530 minutes. Purity = 92%. HPLC conditions: method B. Absolute stereochemistry not determined.

[00473] Example 152(b) Second elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 50-100% B for 20 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 2 (8.1 mg, 7.3%). ESI MS (M+H)+ = 519.1. HPLC peak tr = 2.470 minutes. Purity = 100%. HPLC conditions: method B. Absolute stereochemistry not determined.

[00474] Example 152(c) Third elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 50-100% B for 20 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 3 (13.7 mg, 12%). ESI MS (M+H)+ = 519.1. HPLC peak tr = 2.481 minutes. Purity = 97%. HPLC conditions: method B. Absolute stereochemistry not determined.

[00475] Example 152(d) Fourth elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 50-100% B for 20 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 4 (7.5 mg, 6.7%). ESI MS (M+H)+ = 518.9. HPLC peak tr = 2.361 minutes. Purity = 99%. HPLC conditions: method B. Absolute stereochemistry not determined.Example 153 (±)-2-((trans)-4-((1,8-Naphthyridin-4-yl)oxy)cyclohexyl)-N-(4-chlorophenyl )propanamide Intermediates 153A and 153B. Ethyl 2-(( cis )-4-hydroxycyclohexyl)propanoate (minor) and ethyl 2-(( trans )-4-hydroxycyclohexyl)propanoate (major)

[00476] To a solution of Intermediate 143C (0.241 g, 1.216 mmol) in MeOH (6.08 ml) was added sodium borohydride (0.047 g, 1.240 mmol). The reaction was allowed to stir at RT overnight. The reaction was quenched with water and extracted with EtOAc. The layers were separated. The aqueous phase was extracted with EtOAc (2X). The combined organic phases were dried over Na2SO4, filtered, and concentrated to provide a residue. Purification of raw material by silica gel chromatography using an ISCO machine (40 g column, 40 mL/min, 0-100% EtOAc in hexanes for 19 min, tr = 10.5 minutes = cis, 12 minutes = trans ) provided 153A (173 mg, 71%) and 153B (37 mg, 15%). The biggest product is trans. The smaller product is cis. cis elutes first. Trans elutes second. ESI MS (M+H)+ = 201.1.Intermediate 153C.2-((trans)-4-((1,8-naphthyridin-4-yl)oxy)cyclohexyl)ethyl propanoate

[00477] To a solution of 153B (59.5 mg, 0.297 mmol) in DMF (495 μl) NaH (19.80 mg, 0.495 mmol) was added. After 30 min, 4-bromo-1,8-naphthyridine (51.8 mg, 0.248 mmol) was added. The reaction was heated to 80 °C overnight. The reaction is quenched with a saturated aqueous solution of NH4Cl and diluted with EtOAc. The layers were separated. The aqueous phase was extracted with EtOAc (2X). The combined organic phases were washed with water, dried over Na2SO4, filtered, and concentrated to provide a brown residue. Purification of the crude material by silica gel chromatography using an ISCO machine (40 g column, 40 mL/min, 0-20% MeOH in CH2Cl2 for 9 min, tr = 7.5 min) gave 153C (59.8 mg, 0.182 mmol, 73.5% yield) as a colorless residue. ESI MS (M+H)+ = 329.2. HPLC peak tr = 0.72 minutes. HPLC conditions: method A. Example 153. (±)-2-((trans)-4-((1,8-naphthyridin-4-yl)oxy)cyclohexyl)-N-(4-chlorophenyl)propanamide

[00478] To a solution of 4-chloroaniline (0.080 g, 0.628 mmol) in THF (0.1 mL) at 0 °C was added a solution of isopropylmagnesium chloride (0.314 mL, 0.628 mmol). The resulting solution was warmed to room temperature and stirred for 5 min, then Intermediate 153C (0.0516 g, 0.157 mmol) in THF (0.38 mL) was added dropwise. The reaction was heated to 70 °C for 2 h, then allowed to cool to room temperature. The reaction was quenched with a saturated aqueous solution of NH4Cl and diluted with EtOAc. The layers were separated. The aqueous phase was extracted with EtOAc (3X). The combined organic phases were dried over Na2SO4, filtered, and concentrated to provide a residue. The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 20-60% B for 20 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to provide Example 153 as a racemate (6.7 mg, 10%). ESI MS (M+H)+ = 410.2. HPLC peak tr = 1.803 minutes. Purity = 99%. HPLC conditions: method B. Example 154 N-(4-Chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)butanamide (mixture of four isomers) Intermediate 154A. Ethyl 2-(1,4-dioxaspiro[4,5]decan-8-ylidene)acetate

[00479] To a flask containing sodium hydride (46.1 g, 1153 mmol) THF (1200 mL) was added at 0 °C under nitrogen. Then, triethyl phosphonoacetate (258 g, 1153 mmol) was added dropwise. The reaction mixture was stirred at 0°C for 30 minutes. Then, 1,4-dioxaspiro[4,5]decan-8-one (150 g, 960 mmol) was added and stirred at 0 °C for 2 h. The reaction mixture was warmed to room temperature and stirred for 16 hours. The reaction mixture was quenched with water (500 ml) and the mixture was concentrated in vacuo. The residue was extracted with ethyl acetate (3 x 1000 mL). The combined organic layer was washed with water (500 ml) and brine (500 ml) successively. The filtrate was dried over sodium sulfate, filtered, and concentrated in vacuo. The crude material was purified by flash column chromatography, eluting with 0-30% ethyl acetate in petroleum ether to give Intermediate 154A (light yellow oil, 135 g, 597 mmol, 62.1% yield). ). 1H NMR (400 MHz, chloroform-d) δ: 5.66 (s, 1H), 4.14 (q, J=7.2 Hz, 2H), 4.02 - 3.82 (m, 4H ), 3.24 - 2.86 (m, 2H), 2.63 - 2.27 (m, 2H), 1.98 - 1.68 (m, 4H), 1.27 (t, J=7 .2Hz, 3H).Intermediate 154B. Ethyl 2-(1,4-dioxaspiro[4,5]decan-8-yl)acetate

[00480] Intermediate 154A (13.88 g, 61.3 mmol) was taken up in EtOAc (61.3 ml) and added to a Parr hydrogenation flask containing palladium on carbon (1.306 g, 12.27 mmol) (54 % wt/wt water), 10% Pd/C under a nitrogen atmosphere. A reaction flask was purged with nitrogen, then with hydrogen. After charging the flask with hydrogen to 3.51 kg/cm2 (50 psi), the flask was placed on a Parr shaker and shaken. After 4 hours, the reaction mixture was filtered over pressed CELITE® and concentrated in vacuo to provide Intermediate 154B, ethyl 2-(1,4-dioxaspiro[4,5]decan-8-yl)acetate (colorless oil, 13. 78 g, 60.4 mmol, 98% production). LC-MS Anal. Calculated for C12H20O4 228.14, found [M+H]+ 229.1. Tr = 0.83 minutes (Method A). 1H NMR (400 MHz, chloroform-d) δ: 4.31 - 4.08 (m, 2H), 4.00 - 3.86 (m, 4H), 2.22 (d, J=7.0 Hz , 2H), 1.91 - 1.79 (m, 1H), 1.78 - 1.70 (m, 4H), 1.63 - 1.50 (m, 2H), 1.37 - 1.14 (m, 5H).Intermediate 154C. Ethyl 2-(4-oxocyclohexyl)acetate

[00481] In a 10 liter reactor, Intermediate 154B (67.5 g, 296 mmol) was taken in acetone (5000 mL). To the reaction mixture was added 1 M HCl solution (1183 mL, 1183 mmol) and the resulting mixture was heated under reflux for 2 h. The reaction mixture was concentrated to remove acetone. The residue was extracted with ethyl acetate (3 x 1000 mL). The combined organic layer was washed with water and brine. The organic layer was dried over sodium sulfate and concentrated in vacuo. The crude material was purified by flash column chromatography, eluting with 0-20% ethyl acetate in petroleum ether to give Intermediate 154C (pale yellow liquid, 40 g, 217 mmol, 73.4% yield). . GC-MS Anal. Calculated for C10H16O3, 184.11 found [M+H]+ 184. Tr = 10.03 minutes (Method C).Intermediate 154D. Ethyl 2-(4-(trifluoromethylsulfonyloxy)cyclohex-3-enyl)acetate

[00482] In a 2-liter 4-neck flask, 2,6-di-tert-butyl-4-methylpyridine (84 g, 407 mmol) was taken in dichloromethane (500 mL) under nitrogen. Triflic anhydride (55.0 mL, 326 mmol) was added dropwise. Then, a solution of Intermediate 154C (50 g, 271 mmol) in dichloromethane (500 mL) was added slowly. After completion of the addition, the reaction mixture was stirred at room temperature overnight. The reaction mixture was diluted with 1000 mL of dichloromethane and washed with water and sodium carbonate solution and then water. The organic layer was dried over sodium sulfate and concentrated in vacuo. The crude material was purified by flash column chromatography, eluting with 0-10% ethyl acetate in petroleum ether to provide Intermediate 154D (light yellow oil, 65 g, 206 mmol, 76% yield). GC-MS Anal. Calculated for C11H15F3O5S, 316.06 found [M+H]+ 317. Tr = 10.16 minutes (Method C).Intermediate 154E. Ethyl 2-(4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-enyl)acetate

[00483] In a 2-liter four-neck flask, Intermediate 154D (120 g, 379 mmol), bis(pinacolato)diboron (106 g, 417 mmol), and potassium acetate (112 g, 1138 mmol) were taken in 1,4-dioxane (1200 mL) under nitrogen. Nitrogen was purged into the reaction mixture for 10 minutes. Then, 1,1'-bis(diphenylphosphine)ferrocene-palladium dichloride dichloromethane complex (15.49 g, 18.97 mmol) was added. The reaction mixture was heated at 80 °C for 16 h. The reaction mixture was concentrated. The residue was divided between ethyl acetate and water, filtered through a bed of CELITE®. The organic layer was separated and the aqueous layer was extracted with ethyl acetate (3X). The combined organic layer was washed with water, brine, and dried over sodium sulfate and concentrated in vacuo. The crude material was purified by flash column chromatography, eluting with 0-10% ethyl acetate in petroleum ether to provide Intermediate 154E (light yellow oil, 56 g, 190 mmol, 50.2% yield). GC-MS Anal. Calculated for C16H27BO4, 294.20 found [M+H]+ 295.3. Tr = 1.10 minutes (Method A). 1H NMR (400 MHz, chloroform-d) δ: 6.52 (dd, J=4.1, 1.9 Hz, 1H), 4.14 (q, J=7.1 Hz, 2H), 2, 62 - 1.97 (m, 6H), 1.94 - 1.68 (m, 2H), 1.33 - 1.21 (m, 16H) Intermediate 154F. Ethyl 2-(4-(6-fluoroquinolin-4-yl)cyclohex-3-en-1-yl)acetate

[00484] Ethyl 2-(4-(4,4,5,5-Tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3-en-1-yl)acetate (Intermediate 154E) (5 g, 17.00 mmol) was taken up in dioxane (28.3 ml) and water (7.08 ml). 4-Chloro-6-fluoroquinoline (2.57 g, 14.15 mmol) was added followed by K2CO3 (5.87 g, 42.5 mmol). The mixture was bubbled with nitrogen for 5 minutes before adding Pd(Ph3P)4 (0.327 g, 0.283 mmol). After addition, the reaction was unoccupied and recharged with N2 three times and then sealed (parafilmed sealed vial) and heated to 100 °C for 16 hours. The reaction was concentrated in vacuum and purified directly via silica gel flash column chromatography to provide Intermediate 154F (4.22 g, 13.47 mmol, 95% yield). LC-MS Anal. Calculated for C19H20FNO2 313.15, found [M+H]+ 314.1 Tr = 0.75 minutes (Method A). Intermediate 154G. Ethyl 2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)acetate

[00485] Intermediate 154F (4.22 g, 13.47 mmol) was dissolved in MeOH (67.3 ml) and ammonium formate (4.25 g, 67.3 mmol) was added. The vessel was equipped with a reflux condenser and unoccupied and flooded with nitrogen gas 3 times. Then, palladium on carbon (0.143 g, 1.347 mmol) (Degussa type, wet) was added and the reaction was heated at reflux for 1 hour. The reaction was cooled, concentrated in vacuum, and diluted with DCM. The solids were filtered and the filtrate was concentrated to provide crude Intermediate 154G (4.20 g, 13.32 mmol, 99% yield) as a mixture of cis and trans diastereomers. LC-MS Anal. Calculated for C19H22FNO2 315.16, found [M+H] 316.2 Tr = 0.76 minutes (Method A). Intermediate 154H. Ethyl 2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)butanoate

[00486] To the vial containing THF (6 mL) lithium diisopropylamide (2.0 M solution in THF) (3.17 mL, 6.34 mmol) was added at -78 °C, followed by addition of 1 ,3-dimethyltetrahydropyrimidin-2(1H)-one (0.573 mL, 4.76 mmol) and a solution of intermediate 154G (1.0 g, 3.17 mmol) in THF (10 mL) dropwise at -78° W. The resulting mixture turned brown solution and stirred at -78 °C for 1 h, then iodoethane (0.51 mL, 6.34 mmol) was added slowly. The reaction mixture was then stirred at ice bath temperature for 1 h, warmed to room temperature overnight. The reaction was quenched by pouring into water and extracting with EtOAc. The combined organic layer was washed with brine, dried with MgSO4, filtered and concentrated in vacuo. The residue was dissolved in DCM and purified by silica gel flash chromatography, eluting with 020% ethyl acetate in hexane to provide Intermediate 154H (oil, 0.81 g, 2.359 mmol, 74.4% yield). LC-MS Anal. Calculated for C21H26FNO2, 343.19 found [M+H] 344.3. Tr = 0.87-0.88 minutes (Method A). 1H NMR (400 MHz, chloroform-d) δ: 8.88 - 8.77 (m, 1H), 8.18 - 8.06 (m, 1H), 7.66 (dd, J=10.6, 2.6 Hz, 1H), 7.47 (ddd, J=9.2, 8.0, 2.9 Hz, 1H), 7.36 (d, J=4.6 Hz, 1H), 4, 25 - 4.15 (m, 2H), 3.34 - 3.09 (m, 1H), 2.70 - 2.16 (m, 1H), 2.13 - 1.49 (m, 13H) , 1.36 -1.24 (m, 3H), 1.00 - 0.90 (m, 3H) Intermediate 154I. 2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)butanoic acid

[00487] To a solution of ethyl 2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)butanoate (0.81 g, 2.359 mmol) in THF (4 mL) and MeOH (7 mL) was added solution of 2.0 M LiOH (7.1 mL, 14.2 mmol) slowly. The reaction mixture was stirred at rt overnight. The next day, more LiOH solution (7.1 mL, 14.2 mmol) was added to the reaction mixture and the resulting reaction mixture was heated at 70 °C for 28 h. The reaction mixture was cooled and ethyl acetate was added to the mixture. The aqueous layer was separated and to the aqueous layer was added 1N HCl solution to adjust the pH to 5-6. The resulting mixture was diluted with water and CHCl3:2-propanol (2:1). The organic layer was separated and dried over MgSO4. The filtrate was concentrated in vacuum to provide Intermediate 154I as a mixture of cis and trans isomer (3:2) (0.64 g, 2.029 mmol, 86% yield). LC-MS Anal. Calculated for C19H22FNO2 315.16 found [M+H] 316.3. Tr = 0.72 minutes (Method A). 1H NMR (400 MHz, chloroform-d) δ: 8.83 (d, J=4.4 Hz, 1H), 8.30 - 8.03 (m, 1H), 7.67 (dd, J =10.6, 2.4 Hz, 1H), 7.48 (ddd, J=9.2, 7.9, 2.6 Hz, 1H), 7.38 (d, J=4.6 Hz, 1H), 7.32 - 7.27 (m, 1H), 3.37 - 3.07 (m, 1H), 2.77 - 2.21 (m, 1H), 2.11 - 1.30 ( m, 11H), 1.07 - 1.00 (m, 3H). Example 154: N-(4-Chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)cyclohexyl)butanamide

[00488] To a solution of Intermediate 154I (60 mg, 0.190 mmol) thionyl chloride (0.21 mL, 2.85 mmol) and 1 drop of DMF were added. The reaction mixture was stirred at room temperature for 2 h, and the mixture was diluted with 5 mL of toluene and concentrated in vacuo. The residue was dried under high vacuum for 1 h. To the residue were added acetonitrile (3 mL), 4-chloroaniline (36.4 mg, 0.285 mmol) and 4-methylmorpholine (0.13 mL, 1.14 mmol). The reaction mixture was stirred at room temperature for 0.6 h. The reaction mixture was heated at 70 °C for 1 h. To the reaction mixture was added additional 4-methylmorpholine (0.13 mL, 1.14 mmol) and the resulting reaction mixture was heated at 70 °C overnight. The reaction mixture was diluted with ethyl acetate and brine. The organic layer was separated and concentrated in vacuo. The residue was dissolved in MeOH, filtered, and purified by preparative HPLC to provide Example 154 as a mixture of 4 isomers (7.1 mg, 0.017 mmol, 8.8% yield). LC-MS Anal. Calculated for C25H26ClFN2O, 424.17, found [M+H] 425.3. Tr = 1.72 minutes (Method K). 1H NMR (500 MHz, DMSO-d6) δ: 10.31 - 9.93 (m, 1H), 9.00 - 8.68 (m, 1H), 8.09 (dd, J=8.9, 5.9 Hz, 1H), 7.97 (d, J=9.3 Hz, 1H), 7.66 (d, J=8.7 Hz, 3H), 7.61 - 7.42 (m, 1H), 7.34 (d, J=8.7 Hz, 2H), 3.40 - 3.31 (m, 1H), 2.84 - 2.64 (m, 1H), 2.05 - 1 .15 (m, 11H), 0.96 - 0.70 (m, 3H).Example 155 (R)-N-(4-chlorophenyl)-4-hydroxy-2-((cis)-4-(quinolin -4-yl)cyclohexyl)butanamide Intermediate 155A: (R)-N-(4-chlorophenyl)-4-oxo-2-((cis)-4-(quinolin-4-yl)cyclohexyl)butanamide

[00489] To a solution of Example 224 (0.63 g, 1.5 mmol) in 15 mL of a 3:1 mixture of dioxane and water, NaIO4 (1.28 g, 6 mmol) and 2. 6-lutidine (0.32 g, 3 mmol) to provide a white suspension. OsO4 (5 vol%, 0.15 mL) was then added. After 2 hours, the reaction was completed by TLC, quenched with water, and extracted three times with EtOAc. The combined organics were washed once with brine and then dried over MgSO4. Filtration and concentration gave crude Preparation 155A as a brown oil which was carried away without further purification. Example 155: (R)-N-(4-chlorophenyl)-4-hydroxy-2-((cis)-4-(quinolin-4-yl)cyclohexyl)butanamide

[00490] To a solution of Intermediate 155A (0.16 g, 0.38 mmol) in MeOH (3.8 mL) was added sodium borohydride (0.043 g, 1.14 momol). After 1 hour, the reaction was quenched with saturated aqueous NH4Cl and extracted with a solution of 10% MeOH in EtOAc. Flash chromatography on silica gel provided 2.2 mg of Example 155 as a white solid. MS(ES): m/z = 443.3 [M+H]+. Tr = 2.72 minutes (Method L). Examples 156(a) and 156(b) N-(4-chlorophenyl)-2-((cis)-4-cyano-4-(quinolin-4-yl)cyclohexyl )acetamide and N-(4-chlorophenyl)-2-((trans)-4-cyano-4-(quinolin-4-yl)cyclohexyl)acetamide (relative stereochemistry not determined and arbitrarily specified) Intermediate 156A: 4-oxo-1-(quinolin-4-yl)cyclohexane-1-carbonitrile

[00491] To a solution of 2-(quinolin-4-yl)acetonitrile (2.0 g, 11.4 mmol) in THF (30 mL) was added ethyl acrylate (2.38 g, 23.8 mmol) followed by potassium tert-butoxide (1.6 g, 14.3 mmol) and the mixture was stirred at room temperature. After 1 hour, water (200 mL) was added followed by heating to 85 °C for 18 hours. The reaction was then cooled to room temperature and extracted three times with EtOAc. The combined organics were washed once with brine and dried over MgSO4. Filtration and subsequent concentration provided the crude product as a brown oil. Purification by flash chromatography on silica gel (0100% EtOAc in hexanes) provided Intermediate 156A (800 mg) as a white solid. Intermediate 156B: 2-(4-cyano-4-(quinolin-4-yl)cyclohexylidene )ethyl acetate

[00492] To a solution of ethyl 2-(diethoxyphosphoryl)acetate (0.75 g, 3.36 mmol) in THF (7 mL) was added sodium tert-butoxide (0.32 g, 3.36 mmol) at 0°C. After 10 minutes, a solution of intermediate 156A (0.80 g, 3.2 mmol) in THF (3 mL) was added to the reaction. After another 2 hours, the reaction was quenched with water, extracted three times with EtOAc. The combined organics were washed once with brine, dried over MgSO4 and filtered to provide the crude product. Purification by flash chromatography on silica gel (85% EtO-Ac/hexanes) provided Intermediate 156B (1.0 g) as a white solid. Intermediate 156C: 2-(4-cyano-4-(quinolin-4-yl )cyclohexyl)ethyl acetate

[00493] To a solution of Preparation 156 B (1.00 g, 3.13 mmol) in MeOH (31 mL) was added Pd/C (Degussa type, 0.10 g, 10% Pd). Hydrogen was introduced via a balloon. After 16 hours at RT, the reaction was purged with argon and then filtered through CELITE® rinsing with DCM. Concentration provided raw Preparation 156 C as a mixture of cis and trans isomers which was used without further purification.

[00494] Examples 156(a) and 156(b): N-(4-chlorophenyl)-2-((cis)-4-cyano-4-(quinolin-4-yl)cyclohexyl)acetamide and N-(4 -chlorophenyl)-2-((trans)-4-cyano-4-(quinolin-4-yl)cyclohexyl)acetamide (relative stereochemistry not determined and arbitrarily specified)

[00495] Examples 156A and 156B were prepared by the method described in General Procedure G to provide a mixture of cis and trans isomers. These were separated by flash chromatography on silica gel to give Example 156A and Example 156B.

[00496] Example 156A MS(ES): m/z = 393.2 [M+H]+. Tr = 0.84 minutes (Method M).

[00497] Example 156B MS(ES): m/z = 393.2 [M+H]+. Tr = 2.77 minutes (Method L). Example 157 (R)-N-(4-chlorophenyl)-5-hydroxy-2-((cis)-4-(quinolin-4-yl)cyclohexyl)pentanamide

[00498] To a solution of Example 224 (0.140 g, 0.33 mmol) in THF (3.3 mL) at -78 °C, BH 3-DMS (0.189 mL, 0.37 mmol, solution at 2 .0 M) followed by heating to room temperature. After 2 hours at RT, the solution was cooled to -78 °C and 5 mL of 1 N NaOH and 5 mL of 30% hydrogen peroxide were added followed by warming to room temperature. After 5 hours, quenched with saturated aqueous NH4Cl and extracted three times with EtOAc. The combined organics were washed once with brine and dried over MgSO4. Filtration and concentration provided the raw product. Purification by flash chromatography on silica gel provided Example 157 as a white solid. MS(ES): m/z = 394.2 [M+H]+. Tr = 0.81 minutes (Method M).Example 158 N-(4-chlorophenyl)-2-(1-methoxy-4-(quinolin-4-yl)cyclohexyl)acetamide (simple diastereomer, relative stereochemistry not determined ) Intermediate 158A: ethyl 2-(1-methoxy-4-(quinolin-4-yl)cyclohexyl)acetate

[00499] To a solution of ethyl 2-(1-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetate (0.22 g, 0.69 mmol), prepared by the methods in Example 30, in DME ( 3 mL) at -78°C KHMDS (1.5 mL, 0.5 M in toluene, 0.756 mmol) was added followed by 18-crown-6 (0.20 g, 0.756 mmol). After 30 minutes, MeI (0.057 mL, 0.756 mmol) was added. After a further 30 minutes, the reaction was quenched with saturated aqueous NH4Cl and water, extracted three times with EtOAc, the combined organics washed once with brine, dried over MgSO4, filtered and concentrated to give crude Intermediate 158 A as a single isomer, stereochemistry not confirmed. Example 158 N-(4-chlorophenyl)-2-(1-methoxy-4-(quinolin-4-yl)cyclohexyl)acetamide

[00500] Example 158 was prepared from Intermediate 158A by the methods shown in Example 30. Isolated as a simple isomer, relative stereochemistry not determined. MS(ES): m/z = 379.2 [M+H]+. Tr = 2.29 minutes (Method L).Example 159 N-(4-fluorophenyl)-2-(4-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetamide (simple diastereomer, relative stereochemistry not determined ) Intermediate 159A: ethyl 2-(4-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetate

[00501] To a solution of 4-bromoquinoline (0.36 g, 1.75 mmol) in THF (8 mL) at -78 °C was added tert-butyl lithium (1.7 M solution, 2.1 mL , 3.51 mmol). After 5 minutes, a solution of ethyl 2-(4-oxocyclohexyl)acetate (0.294 g, 1.60 mmol) in THF (2 mL) was added. After 1 hour, 1N NaOH was added followed by warming to room temperature. The mixture was then extracted three times with EtOAc and the combined organics washed once with brine. The organics were then dried over MgSO4, filtered and concentrated to provide the crude product. Purified by flash chromatography on silica gel eluting with 0 - 100% EtOAc in hexanes to give Intermediate 159A (214 mg) as a yellow oil. Example 159: N-(4-fluorophenyl)-2-(4-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetamide (simple diastereomer, relative stereochemistry not determined)

[00502] Example 159 was prepared from Intermediate 159A and 4-fluoroaniline by the methods described in General Method G. Example 159 was isolated as a single isomer under crystallization after concentration, stereochemistry not confirmed. MS(ES): m/z = 393.2[M+H]+. Tr = 2.77 minutes (Method L). Example 160 N-(4-chlorophenyl)-2-(4-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetamide (simple diastereomer, relative stereochemistry not determined) Example 160: N-(4-chlorophenyl)-2-(4-hydroxy-4-(quinolin-4-yl)cyclohexyl)acetamide

[00503] Example 160 was prepared from Preparation 159A and 4-chloroaniline by the methods described in General Method G. Example 160 was isolated as a simple isomer under crystallization after concentration, stereochemistry not confirmed. MS(ES): m/z = 431.3 [M+H]+. Tr = 0.85 minutes (Method M). Table 7. Examples 161-218 prepared by the methods described previously Table 8. Examples 220-228 prepared by the methods described previously Examples 229 and 230 Example 229: N-(4-chlorophenyl)-2-( trans -4-(6-fluoroquinolin-4-yl)-1-hydroxycyclohexyl)propanamide Example 230: N-(4-chlorophenyl) -2-( trans -4-(6-fluoroquinolin-4-yl)-1-hydroxycyclohexyl)propanamide (absolute stereochemistry unknown) 229A: Methyl 2-( trans -4-(6-fluoroquinolin-4-yl)-1-hydroxycyclohexyl)propanoate 229B: Methyl 2-(cis-4-(6-fluoroquinolin-4-yl)-1-hydroxycyclohexyl)propanoate

[00504] A solution of 4-(6-fluoroquinolin-4-yl)cyclohexanone (200 mg, 0.822 mmol), methyl 2-bromopropanoate (275 mg, 1.644 mmol) in CH3CN (6 mL) at 0 °C was Tris(triphenylphosphine)rhodium(I) chloride (45.6 mg, 0.049 mmol) was added. The reaction was stirred at 0 °C for 30 min, then diethylzinc (1.0 M solution in heptane) (1.726 mL, 1.726 mmol) was added. The reaction was stirred at room temperature for 16 h. The reaction was diluted with EtOAc and saturated NH4Cl. The organic was separated and washed with brine, dried over MgSO4, filtered and concentrated to provide a crude material. This crude material was purified with ISCO 24 g, 40 mL/min. 0100% EtOAc/hexane in 50 min. Product 229A (87 mg, 0.26 mmol, 32%) was eluted with 50% EtOAc/hexane. Product 229B (122 mg, 0.364 mmol, 44%) was eluted with 60% EtOAc/hexane.

[00505] 229A: 1H NMR (400 MHz, CHLOROFORM-d) β 8.83 (d,J=4.6 Hz, 1H), 8.14 (dd, J=9.2, 5.7 Hz, 1H ), 7.67 (dd, J=10.5, 2.8 Hz, 1H), 7.50 (ddd, J=9.2, 8.0, 2.8 Hz, 1H), 7.33 ( d, J=4.6 Hz, 1H), 3.78 (s, 3H), 3.65 (s, 1H), 3.36 - 3.23 (m, 1H), 3.08 (q, J =7.1 Hz, 1H), 2.18 - 2.08 (m, 1H), 2.06 - 1.94 (m, 2H), 1.94 - 1.81 (m, 2H), 1, 79 - 1.68 (m, 3H), 1.68 - 1.51 (m, 1H), 1.36 - 1.23 (m, 3H) LC-MS: M+H=332.2 (tr= 0.59 min) (Method A)

[00506] 229B: 1H NMR (400 MHz, CHLOROFORM-d) β 8.84 (d, J=4.6 Hz, 1H), 8.14 (dd, J=9.2, 5.7 Hz, 1H ), 7.68 (dd, J=10.6, 2.8 Hz, 1H), 7.49 (ddd, J=9.1, 8.0, 2.8 Hz, 1H), 7.39 ( d, J=4.6 Hz, 1H), 3.78 (s, 3H), 3.21 - 3.04 (m, 2H), 2.56 (q, J=7.2 Hz, 1H), 2.18 - 1.97 (m, 3H), 1.93 - 1.67 (m, 5H), 1.65 (s, 2H), 1.55 - 1.41 (m, 1H), 1, 36 - 1.23 (m,3H) Examples 229 and 230

[00507] To a solution of 4-chloroaniline (63.5 mg, 0.498 mmol) in THF (1 mL) at RT was added iPrMgCl (2.0 M in THF) (0.415 mL, 0.830 mmol) dropwise. Bubbles have evolved. The reaction was stirred at room temperature for 5 min, then 229A (55 mg, 0.166 mmol) in THF (0.3 mL) was added. The reaction was stirred at 70 °C for 1 h. The reaction was diluted with saturated NH4Cl and EtOAc. The organic was separated and washed with brine, dried over MgSO4, filtered and concentrated to dryness. The crude material was purified with a 24 g, 35 mL/min ISCO column. 0-100% EtOAc/hexane in 30 min. The desired product was eluted with 80% EtOAc/hexane to provide N-(4-chlorophenyl)-2-(trans-4-(6-fluoroquinolin-4-yl)-1-hydroxycyclohexyl)propanamide (58 mg, 0.135 mmol , 81% production) as white solid.

[00508] The racemate was purified through preparative SFC with the following conditions: Column: Chiral OD-H 25 x 3 cm ID, 5 μm particles; Mobile phase A: 70/30 CO2/MeOH; Detector wavelength: 220 nm; Flow: 100 mL/min. The fractions ("Peak-1" tr = 3.98 minutes (Example 229) and "Peak-2" tr = 4.99 minutes (Example 230));

[00509] Example 229: 1H NMR (400 MHz, CHLOROFORM-d) β 8.83 (d, J=4.6 Hz, 1H), 8.14 (dd, J=9.2, 5.7 Hz, 1H), 7.67 (dd, J=10.5, 2.8 Hz, 1H), 7.50 (ddd, J=9.2, 8.0, 2.8 Hz, 1H), 7.33 (d, J=4.6 Hz, 1H), 3.78 (s, 3H), 3.65 (s, 1H), 3.36 - 3.23 (m, 1H), 3.08 (q, J=7.1 Hz, 1H), 2.18 - 2.08 (m, 1H), 2.06 - 1.94 (m, 2H), 1.94 - 1.81 (m, 2H), 1 .79 - 1.68 (m, 3H), 1.68 - 1.51 (m, 1H), 1.36 - 1.23 (m, 3H) LC-MS: M+H=332.2 (tr =0.59 min)

[00510] Example 230: 1H NMR (400 MHz, CHLOROFORM-d) β 8.83 (d, J=4.6 Hz, 1H), 8.14 (dd, J=9.2, 5.7 Hz, 1H), 7.67 (dd, J=10.5, 2.8 Hz, 1H), 7.50 (ddd, J=9.2, 8.0, 2.8 Hz, 1H), 7.33 (d, J=4.6 Hz, 1H),3.78 (s, 3H), 3.65 (s, 1H), 3.36 - 3.23 (m, 1H), 3.08 (q, J=7.1 Hz, 1H), 2.18 - 2.08 (m, 1H), 2.06 - 1.94 (m, 2H), 1.94 - 1.81 (m, 2H), 1 .79 - 1.68 (m, 3H), 1.68 - 1.51 (m, 1H), 1.36 - 1.23 (m, 3H) LC-MS: M+H=332.2 (tr =0.59 min) (Method A) Example 231 and 232 Example 231: N-(4-chlorophenyl)-2-( cis -4-(6-fluoroquinolin-4-yl)-1-hydroxycyclohexyl)propanamide Example 232: N-(4-chlorophenyl)-2-( cis -4-(6-fluoroquinolin-4-yl)-1-hydroxycyclohexyl)propanamide (absolute stereochemistry unknown)

[00511] To a solution of 4-chloroaniline (61.2 mg, 0.480 mmol) in THF (2 mL) at RT was added iPrMgCl (2.0 M in THF) (0.400 mL, 0.800 mmol) dropwise. Bubbles have evolved. The mixture was stirred at room temperature for 5 min, then Preparation 229B (53 mg, 0.160 mmol) was added. The reaction was heated to 70 °C for 1 h, cooled to room temperature. The mixture was diluted with saturated NH4Cl and EtOAc. The organic was separated and washed with brine, dried over MgSO4, filtered and concentrated to dryness. The crude material was purified with a 24 g, 35 mL/min ISCO column. 0-100% EtOAc/hexane in 45 min. The desired product was eluted with 75% EtOAc/hexane to provide N-(4-chlorophenyl)-2-((1s,4s)-4-(6-fluoroquinolin-4-yl)-1-hydroxycyclohexyl)propanamide (55 mg, 0.128 mmol, 80% production) as white solid. The racemate was purified by preparative SFC with the following conditions: Column: Chiral OD-H 25 x 3 cm ID, 5 μm particles; Mobile phase A: 70/30 CO2/MeOH; Detector wavelength: 220 nm; Flow: 100 mL/min. The fractions ("Peak-1" tr = 4.58 minutes (Example 231) and "Peak-2" tr = 5.33 minutes (Example 232));

[00512] Example 231: 1H NMR (400 MHz, CHLOROFORM-d) β 8.83 (d, J=4.5 Hz, 1H), 8.14 (dd, J=9.2, 5.7 Hz, 1H), 7.94 (s, 1H), 7.66 (dd, J=10.6, 2.8 Hz, 1H), 7.55 - 7.45 (m, 3H), 7.38 (d , J=4.6 Hz, 1H), 7.35 - 7.31 (m, 2H), 3.60 (br. s., 1H), 3.24 - 3.02 (m, 1H), 2 .38 (q, J=7.1 Hz, 1H), 2.21 - 1.95 (m, 3H), 1.95 - 1.83 (m, 2H), 1.80 - 1.64 (m , 2H), 1.49 (td, J=13.3, 4.1 Hz, 1H), 1.42 (d, J=7.1 Hz, 3H) LC-MS: M+H=332.2 (tr=0.78 min) (Method A)

[00513] Example 232: 1H NMR (400 MHz, CHLOROFORM-d) β 8.83 (d, J=4.5 Hz, 1H), 8.14 (dd, J=9.2, 5.7 Hz, 1H), 7.94 (s, 1H), 7.66 (dd, J=10.6, 2.8 Hz, 1H), 7.55 - 7.45 (m, 3H), 7.38 (d , J=4.6 Hz, 1H), 7.35 - 7.31 (m, 2H), 3.60 (br. s., 1H), 3.24 - 3.02 (m, 1H), 2 .38 (q, J=7.1 Hz, 1H), 2.21 - 1.95 (m, 3H), 1.95 - 1.83 (m, 2H), 1.80 - 1.64 (m , 2H), 1.49 (td, J=13.3, 4.1 Hz, 1H), 1.42 (d, J=7.1 Hz, 3H) LC-MS: M+H=332.2 (tr=0.78 min) (Method A) Example 233 (+/-)-N-(4-chlorophenyl)-2-(trans -1-fluoro-4-(6-fluoroquinolin-4-yl)cyclohexyl) propanamide

[00514] To a solution of 229B (20 mg, 0.047 mmol) in CH2Cl2 (1 mL) at RT was added diethylaminosulfur trifluoride (0.019 mL, 0.141 mmol). The reaction was stirred at room temperature for 3 h. The reaction was diluted with water and EtOAc. The organic was separated and washed with brine, dried over MgSO4, filtered and concentrated to dryness. The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 0.1% trifluoroacetic acid; Mobile phase B: 95:5 acetonitrile: water with 0.1% trifluoroacetic acid; Gradient: 25-50% B for 25 minutes, then a 2-minute hold at 50% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was also purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 50-75% B for 25 minutes, then a 2-minute hold at 75% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The yield of Example 233 was 0.5 mg (1.17 mmol, 2.5%). 1H NMR (500 MHz, DMSO-d6) δ 10.13 (s, 1H), 8.84 (d, J=4.4 Hz, 1H), 8.15 - 8.00 (m, 2H), 7 .72 - 7.55 (m, 4H), 7.36 (d, J=8.7 Hz, 2H), 3.21 - 3.12 (m, 1H), 2.15 (br. s., 1H), 2.06 (br. s., 1H), 1.94 (d, J=9.0 Hz, 3H), 1.88 (br. s., 2H), 1.66 (d, J =11.4 Hz, 1H), 1.24 (d, J=6.9 Hz, 3H) LC-MS: M+H = 429.0 (tr=0.82 min) (Method A) Examples 234, 235, 236, 237 Example 234: N-(4-chlorophenyl)-2-(trans -4-(6-fluoroquinolin-4-yl)-4-hydroxycyclohexyl)propanamide (unknown absolute stereochemistry) Example 235: N-(4 -chlorophenyl)-2-(trans -4-(6-fluoroquinolin-4-yl)-4-hydroxycyclohexyl)propanamide (absolute stereochemistry unknown) Example 236: N-(4-chlorophenyl)-2-( cis -4-(6-fluoroquinolin-4-yl)-4-hydroxycyclohexyl)propanamide (unknown absolute stereochemistry) Example 237: N-(4-chlorophenyl)-2 -( cis -4-(6-fluoroquinolin-4-yl)-4-hydroxycyclohexyl)propanamide (absolute stereochemistry unknown) 234A: Ethyl 2-(4-(6-fluoroquinolin-4-yl)-4-hydroxycyclohexyl)propionate

[00515] To a solution of 4-bromo-6-fluoroquinoline (163 mg, 0.721 mmol) in THF (5 mL) at -78 °C was added t-BuLi (1.7M-3.2M in heptane) (0.849 mL, 1.443 mmol) dropwise. It went from light brown to dark brown. The reaction was stirred at -78 °C for 3 min, then ethyl 2-(4-oxocyclohexyl)propanoate (130 mg, 0.656 mmol) in THF (1 mL) was added dropwise. The reaction was stirred at -78 °C for 1 h. The reaction was diluted with saturated NH 4Cl and EtO-Ac. The organic was separated and washed with brine, dried over MgSO4, filtered and concentrated to provide a crude material. This material was purified with 40 g ISCO, 40 mL/min. 0-100% EtO-Ac/hexane in 35 min. The desired product was eluted with 55% EtOAc/hexane to provide 234A (100 mg, 0.290 mmol, 44%) as a mixture of the diastereomers. 234B:N-(4-chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)-4-hydroxycyclohexyl)propanamide

[00516] To a solution of 4-chloroaniline (111 mg, 0.869 mmol) in THF (2 mL) at RT was added iPrMgBr (2.0 M in THF) (0.724 mL, 1.448 mmol) dropwise. Bubbles have evolved. The reaction was stirred at room temperature for 5 min, then 234A (100 mg, 0.29 mmol) in THF (1 mL) was added. The reaction was stirred at 70 °C for 1 h, then cooled to RT and diluted with saturated NH4Cl and EtOAc. The organic was separated and washed with brine, dried over MgSO4, filtered and concentrated to dryness. The crude material was purified with a 24 g, 35 mL/min ISCO column. 0-100% EtOAc/hexane in 30 min. The desired product was eluted with 80% EtOAc/hexane to provide 234B (110 mg, 0.258 mmol, 89%) as a mixture of diastereomers.

[00517] A mixture of 234B diastereomers was purified through preparative SFC with the following conditions: Column: Whelk- O R, R Kromasil 25 x 3 cm ID, 5 μm particles; Mobile phase A: 75/25 CO2/MeOH; Detector wavelength: 220 nm; Flow: 100 mL/min. The fractions ("Peak-1" tr = 9.94 minutes (Example 234) and "Peak-2" tr = 11.49 minutes (Example 236)); "Peak-3" tr = 13.23 minutes (Example 235) and "Peak-4" tr = 14.63 minutes (Example 237);

[00518] Examples 234 and 235: 5 mg each (0.011 mmol, 4.44%) 1H NMR (400 MHz, CHLOROFORM-d) β 8.86 - 8.74 (m, 1H), 8.53 (dd, J=11.7, 2.8 Hz, 1H), 8.13 (dd, J=9.2, 5.9 Hz, 1H), 7.57 - 7.39 (m, 4H), 7.35 - 7.26 (m, 2H), 7.22 (s, 1H), 2.59 (d, J=7.3 Hz, 2H), 2.27 - 2.16 (m, 1H), 2, 09 - 1.86 (m, 5H), 1.38 - 1.18 (m, 6H); LC-MS: M+H=427.1 (tr=0.78 min) (Method A)

[00519] Examples 236 and 237: 40 mg each (0.093 mmol, 36.2%) 1H NMR (400 MHz, CHLOROFORM-d) β 8.83 (d, J=4.6 Hz, 1H), 8.52 (dd, J=11.7, 2.8 Hz, 1H), 8.14 (dd, J=9.3, 6.0 Hz, 1H), 7.58 - 7.53 (m, J=8 .8 Hz, 2H), 7.49 (ddd, J=9.2, 7.6, 2.8 Hz, 1H), 7.42 (d, J=4.5 Hz, 1H), 7.39 - 7.30 (m, 2H), 7.20 (s, 1H), 2.38 - 2.16 (m, 3H), 2.09 - 1.91 (m, 3H), 1.88 - 1 .71 (m, 5H), 1.40 - 1.30 (m, 3H); LC-MS: M+H=427.1 (tr=0.78 min) (Method A)

[00520] Examples 238 - 241 were obtained following the procedures in Example 58 using the corresponding acids and anilines. Example 242 2-(1-(6-Fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)-N-(1-methylcyclohexyl)acetamide 242A. Methyl 2-(4-methylpiperidin-4-yl)acetate

[00521] To a flask charged with MeOH (7.5 mL), at 0 °C under a nitrogen atmosphere, acetyl chloride (1.1 mL, 15.2 mmol) was slowly added. After the addition was complete, the mixture was stirred at 0°C for 5 minutes before a homogeneous mixture of 2-(4-methylpiperidin-4-yl)acetic acid, HCl (675.0 mg, 3.5 mmol) in MeOH (1.5 mL) will be added slowly dropwise. The resulting homogeneous mixture was stirred at 0°C for 5 minutes, then at 60°C for 8 hours, before being concentrated in vacuo to provide the HCl salt of the title compound as a white solid (718.0 mg; 99% production) that was used without further purification. 1H NMR (400 MHz, DMSO-d6) β 9.41 - 9.12 (m, 1H), 3.60 (s, 3H), 3.25 - 3.15 (m, 2H), 2.93 - 2.82 (m, 2H), 2.39 - 2.30 (m, 2H), 1.74 - 1.64 (m, 4H), 1.02 (s, 3H).242B. Methyl 2-(1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)acetate

[00522] To a homogeneous mixture of 4-chloro-6-fluoroquinoline (350.0 mg, 1.9 mmol) in anhydrous NMP (5 mL), in a sealable vial, was added the HCl salt of 2- Methyl (4-methylpiperidin-4-yl)acetate (242A, 480.0 mg, 2.3 mmol) followed by DIPEA (1.6 mL, 9.2 mmol). The vial was sealed and the mixture was stirred at 120°C. After 26 hours, the reaction mixture was cooled to room temperature, then partitioned between water and EtOAc. The layers were separated and the aqueous layer was extracted once again with EtOAc. The organic layers were combined, washed with brine, then concentrated in vacuo to provide the crude product. Purification by Isco chromatography provided methyl 2-(1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)acetate as an oil (565.8 mg; 93% yield). MS (ES): m/z = 317 [M+H]+. tR = 0.66 minutes (Method A). 1H NMR (400 MHz, DMSO-d6) β 8.38 (d, J=5.4 Hz, 1H), 7.96 (dd, J=11.7, 2.8 Hz, 1H), 7.89 - 7.84 (m, 1H), 7.55 - 7.49 (m, 1H), 6.54 (d, J=5.5 Hz, 1H), 3.82 - 3.63 (m, 2H ), 3.59 (s, 3H), 3.54 - 3.34 (m, 2H), 2.45 - 2.38 (m, 2H), 1.87 - 1.72 (m, 4H), 1.05 (s, 3H). 242C. 2-(1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)acetic acid

[00523] To a homogeneous mixture of methyl 2-(1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)acetate (321.0 mg, 1.0 mmol) in MeOH (5 mL ), under a nitrogen atmosphere, 2 M NaOH aqueous solution (1 mL, 2.0 mmol) was added dropwise. The reaction was then stirred at room temperature for 20 hours before being treated with 1 N HCl (aq) to pH 6 on pH test strips. The mixture was then partitioned between water and EtOAc, the layers were separated and the aqueous layer was extracted twice with EtOAc. The aqueous extraction layer was freeze-dried to provide the crude product as an off-white solid (302.1 mg, 98% yield) that was used without further purification. MS (ES): m/z = 303 [M+H]+. tR = 0.58 minutes (Method A). 1H NMR (400 MHz, DMSO-d6) β 12.12 (br.s, 1H), 8.41 (d, J=6.1 Hz, 1H), 8.14 - 8.08 (m, 1H) , 8.00 (dd, J=9.3, 5.7 Hz, 1H), 7.75 - 7.64 (m, 1H), 6.64 (d, J=6.2 Hz, 1H), 3.98 - 3.87 (m, 1H), 3.87 - 3.78 (m, 1H), 3.69 - 3.49 (m, 2H), 2.38 - 2.29 (m, 2H) ), 1.92 - 1.70 (m, 4H), 1.06 (s, 3H). Example 242: 2-(1-(6-Fluoroquinolin-4-yl)-4-methylpiperidin-4-yl) -N-(1-methylcyclohexyl)acetamide

[00524] To the mixture of 2-(1-(6-fluoroquinolin-4-yl)-4-methylpiperidin-4-yl)acetic acid (26.5 mg, 0.09 mmol) in anhydrous DMF (1 mL), In a sealable vial, PyBOP (45.6 mg, 0.09 mmol) was added followed by DIPEA (0.06 mL, 0.34 mmol). The mixture was stirred for 15 minutes before 1-methylcyclohexanamine, HCl (15.7 mg, 0.11 mmol) was added and the vial was sealed. The reaction was stirred at room temperature for 21 hours, before being diluted with DMF, passed through a syringe filter, then purified by preparative HPLC/MS to provide the title compound (17.2 mg; 38% production). MS (ES): m/z = 398 [M+H]+. tR = 1.61 minutes (Method B). 1H NMR (500 MHz, DMSO-d6) δ 8.38 (d, J=5.6 Hz, 1H), 8.02 (d, J=9.7 Hz, 1H), 7.90 - 7.84 (m, 1H), 7.60 (t, J=7.6 Hz, 1H), 7.19 (s, 1H), 6.57 (d, J=5.8 Hz, 1H), 3.94 - 3.64 (m, 2H), 2.16 (t, J=7.8 Hz, 2H), 2.02 - 1.92 (m, 2H), 1.88 - 1.73 (m, 2H ), 1.67 (t, J=7.3 Hz, 2H), 1.47 - 1.31 (m, 5H), 1.30 - 1.10 (m, 8H), 1.04 (s, 3H). Example 243 (±)-N-(4-Chlorophenyl)-2-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)butanamide 243A. tert-butyl 4-(1-Ethoxy-1-oxobutan-2-ylidene)piperidine-1-carboxylate

[00525] To a suspension of NaH (0.29 g, 7.18 mmol) in anhydrous THF (10 mL), under a nitrogen atmosphere, triethyl 2-phosphonobutyrate (1.81 g, 7.18 mmol) was added. mmol) for 5 minutes. The resulting mixture was stirred at room temperature for 20 minutes, during which time it became homogeneous. To this solution was added dropwise a homogeneous mixture of 1-Boc-4-piperidone (1.10 g, 5.52 mmol) in anhydrous THF (2.5 mL). After stirring for 1.5 hours, the reaction was quenched with saturated NH4Cl solution (aq) before being fully extracted with EtOAc. The organic fractions were combined, washed with brine, dried (MgSO4), filtered and concentrated in vacuo to give tert-butyl 4-(1-ethoxy-1-oxobutan-2-ylidene)piperidine-1-carboxylate as clear oil ( 1.64 g; 100% production) which was used without further purification. 1H NMR (400 MHz, DMSO-d6) δ 4.13 (q, J=7.1 Hz, 2H), 3.39 - 3.35 (m, 2H), 3.35 - 3.31 (m, 2H), 2.45 - 2.39 (m, 2H), 2.31 - 2.22 (m, 4H), 1.40 (s, 9H), 1.21 (t, J=7.2 Hz , 3H), 0.93 (t, J=7.5 Hz, 3H). 243B. tert-butyl 4-(1-Ethoxy-1-oxobutan-2-yl)piperidine-1-carboxylate

[00526] A flask loaded with tert-butyl 4-(1-ethoxy-1-oxobutan-2-ylidene)piperidine-1-carboxylate (1.64 g, 5.52 mmol) and under a nitrogen atmosphere was Platinum(IV) oxide (0.07 g, 0.31 mmol) was added followed by careful addition of EtOH (5 mL). The nitrogen line was then replaced with a hydrogen balloon and the mixture was stirred at room temperature. After 15 hours, the reaction mixture was purged with nitrogen before being filtered through a CELITE® pad. The pad was thoroughly rinsed with EtOAc before the combined filtrates were concentrated in vacuo to provide tert-butyl 4-(1-ethoxy-1-oxobutan-2-yl)piperidine-1-carboxylate as a clear oil (1.65 g ; 100% production) that was used without further purification. 1H NMR (400 MHz, DMSO-d6) δ 4.08 (q, J=7.1 Hz, 2H), 3.98 - 3.83 (m, 2H), 2.78 - 2.52 (m, 2H), 2.13 - 2.03 (m, 1H), 1.69 - 1.62 (m, 1H), 1.52 - 1.43 (m, 2H), 1.38 (s, 9H) , 1.28 - 1.13 (m, 5H), 1.10 - 0.98 (m, 2H), 0.80 (t, J=7.3 Hz, 3H).243C. Ethyl 2-(Piperidin-4-yl)butanoate

[00527] To a homogeneous mixture of tert-butyl 4-(1-ethoxy-1-oxobutan-2-yl)piperidine-1-carboxylate (1.65 g, 5.52 mmol) in anhydrous dioxane (5 mL) , under a nitrogen atmosphere, HCl (4 N in dioxane, 10 mL, 40.0 mmol) was added. The mixture was stirred at room temperature for two hours before being concentrated in vacuo to remove volatiles. The resulting oil was treated with saturated aqueous NaHCO3 solution to pH5 and then 1N NaOH (aq) to pH8, before the mixture was fully extracted with EtOAc. The layers were separated and the aqueous layer was lyophilized to provide the title compound as a light yellow solid (1.20 g; 92% yield) which was used in the next step without further purification. MS (ES): m/z = 200 [M+H]+. tR = 0.57 minutes (Method A). 1H NMR (500 MHz, DMSO-d6) δ 4.07 (q, J=7.2 Hz, 2H), 3.01 - 2.78 (m, 2H), 2.48 - 2.29 (m, 3H), 2.07 - 1.98 (m, 1H), 1.60 - 1.35 (m, 5H), 1.18 (t, J=7.1 Hz, 3H), 1.11 - 0 .88 (m, 2H), 0.80 (t, J=7.4 Hz, 3H).243D. Ethyl 2-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)butanoate

[00528] To a homogeneous mixture of 4-chloro-6-fluoroquinoline (365.0 mg, 2.01 mmol) in anhydrous NMP (5 mL), in a sealable vial, was added 2-(piperidin-4-yl) ethyl butanoate (243C, 544.0 mg, 2.31 mmol) followed by DIPEA (1.6 mL, 9.16 mmol). The vial was sealed and the mixture was stirred at 120°C for three hours before being allowed to cool to room temperature. After stirring for 7 days, the mixture was heated to 120°C for three days, then allowed to cool to room temperature, before being partitioned between water and EtOAc. The layers were separated and the aqueous layer was extracted once again with EtOAc. This organic extract was combined with the original organic layer and washed with water, then concentrated in vacuum to provide a dark brown residue. Purification by Isco chromatography provided ethyl 2-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)butanoate as a golden oil (76.1 mg; 11% yield). MS (ES): m/z = 345 [M+H]+. tR = 0.77 minutes (Method A). 1H NMR (400 MHz, CHLOROFORM-d) δ 8.68 (d, J=5.0 Hz, 1H), 8.06 - 8.03 (m, 1H), 7.59 - 7.55 (m, 1H), 7.43 - 7.38 (m, 1H), 6.84 (d, J=4.9 Hz, 1H), 4.21 (q, J=7.1 Hz, 2H), 3, 62 - 3.52 (m, 2H), 2.84 - 2.71 (m, 2H), 2.29 - 2.20 (m, 1H), 2.01 - 1.93 (m, 1H), 1.82 - 1.60 (m, 6H), 1.31 (t, J=7.2 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H).243E. 2-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)butanoic acid

[00529] To a homogeneous mixture of ethyl 2-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)butanoate (76.1 mg, 0.22 mmol) in EtOH (4 mL), under nitrogen atmosphere, NaOH (2 M aqueous solution, 0.2 mL, 0.40 mmol) was added. The mixture was stirred at room temperature for 21 hours before NaOH (2M aqueous solution, 0.2 mL, 0.40 mmol) was added and stirring continued. After 22 hours, NaOH (2 M aqueous solution, 0.2 mL, 0.40 mmol) was added and the reaction was heated to 40 °C. The reaction was stirred for 4 days before NaOH (2M aqueous solution, 0.2 mL, 0.40 mmol) was added and stirring was continued for 21 hours. After cooling to room temperature, the reaction was quenched with 4N HCl in dioxane (to pH 5-6 on pH test strips), stirred for 5 minutes at room temperature, then concentrated in vacuo to provide the crude product as a light yellow solid, which was used in the next step without further purification. MS (ES): m/z = 317 [M+H]+. tR = 0.63 minutes (Method A). Example 243: (±)-N-(4-Chlorophenyl)-2-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)-butanamide

[00530] To the mixture of 2-(1-(6-fluoroquinolin-4-yl)piperidin-4-yl)butanoic acid (35.0 mg, 0.11 mmol) and 4-chloroaniline (17.0 mg, 0 .13 mmol) in anhydrous DMF under nitrogen atmosphere, DIPEA (0.1 mL, 0.57 mmol) was added followed by PyBOP (57.6 mg, 0.11 mmol). The resulting mixture was stirred at room temperature for 98 hours before being diluted with DMF, passed through a syringe filter, then purified by preparative HPLC/MS to provide the title compound (2.5 mg; 3% production). MS (ES): m/z = 426 [M+H]+. tR = 2.22 minutes (Method B). 1H NMR (500 MHz, DMSO-d6) δ 10.15 (s, 1H), 8.64 (d, J=4.9 Hz, 1H), 8.00 (dd, J=9.0, 5, 7 Hz, 1H), 7.67 (d, J=8.7 Hz, 2H), 7.64 - 7.51 (m, 2H), 7.35 (d, J=8.7 Hz, 2H) , 7.07 - 6.96 (m, J=4.9 Hz, 1H), 3.52 - 3.43 (m, 1H), 2.84 - 2.66 (m, 2H), 2.55 - 2.53 (m, 1H), 2.27 - 2.18 (m, 1H), 2.02 - 1.92 (m, J=11.9 Hz, 1H), 1.78 - 1.43 (m, 6H), 0.86 (t, J=7.2 Hz, 3H).Example 244 (±)-N-(4-Chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl) piperazin-1-yl)pentanamide 244A. tert-butyl 4-(6-Fluoroquinolin-4-yl)piperazine-1-carboxylate

[00531] To the mixture of 4-chloro-6-fluoroquinoline (500.0 mg, 2.8 mmol) in anhydrous NMP (5 mL), in a sealable vial, was added 1-Boc-piperazine (750.0 mg, 4.0 mmol) followed by DIPEA (2.0 mL, 11.5 mmol). The vial was capped and the mixture was stirred at 120°C for 15.5 hours. The reaction mixture was cooled to room temperature before being partitioned between water and Et2O. The layers were separated and the aqueous layer was extracted twice more with Et2O. These organic extracts were combined with the original organic layer and washed with brine, dried (Na2SO4), filtered and concentrated in vacuo to provide the crude product. Purification by Isco chromatography provided tert-butyl 4-(6-fluoroquinolin-4-yl)piperazine-1-carboxylate as an oil (719.3 mg; 77% yield). MS (ES): m/z = 332 [M+H]+. tR = 0.70 minutes (Method A). 1H NMR (400 MHz, DMSO-d6) β 8.70 (d, J=4.9 Hz, 1H), 8.09 - 8.00 (m, 1H), 7.77 - 7.67 (m, 1H), 7.67 - 7.62 (m, 1H), 7.07 (d, J=4.9 Hz, 1H), 3.67 - 3.57 (m, 4H), 3.15 - 3 .06 (m, 4H), 1.44 (s, 9H).244B. 6-Fluoro-4-(piperazin-1-yl)quinoline

[00532] A homogeneous mixture of tert-butyl 4-(6-fluoroquinolin-4-yl)piperazine-1-carboxylate (600.0 mg, 1.8 mmol) in dioxane (4 mL), under a nitrogen atmosphere , 4 M HCl in dioxane (10 mL, 40.0 mmol) was added. The resulting mixture was stirred at room temperature for 2.5 hours, during which time a precipitate formed. The heterogeneous mixture was concentrated to approximately 1/2 of its original volume. Vacuum filtration provided the HCl salt of the title compound as an off-white solid (490.0 mg; 100% production) which was used without further purification. MS (ES): m/z = 232 [M+H]+. tR = 0.38 minutes (Method A).244C. (±)-ethyl 2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)pentanoate

[00533] To a heterogeneous mixture of 6-fluoro-4-(piperazin-1-yl)quinoline HCl salt (244B, 200.0 mg, 0.75 mmol) in anhydrous DMF (5 mL), in To a sealable vial, K2CO3 (288.0 mg, 2.08 mmol) was added followed by ethyl 2-bromovalerate (234.0 mg, 1.12 mmol). The vial was then sealed and the mixture was stirred at 60°C. After 16 hours, the reaction was cooled to room temperature, then partitioned between water and EtOAc. The layers were separated and the aqueous layer was extracted once again with EtOAc. The organic layers were combined, washed with brine, dried (anhydrous Na2SO4), filtered and concentrated in vacuo to give the product as a yellow oil which was used in the next step without further purification. MS (ES): m/z = 360 [M+H]+. tR = 0.62 minutes (Method A).244D. (±)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)pentanoic acid

[00534] A homogeneous mixture of ethyl 2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)propanoate (244C, 0.75 mmol) in MeOH (4 mL), under a nitrogen atmosphere , 2M NaOH (aq) (0.8 mL, 1.6 mmol) was added dropwise. The resulting mixture was stirred at room temperature for 64 hours before 2M NaOH (aq) (0.8 mL, 1.6 mmol) was added. After 6 hours of stirring, 2M NaOH (aq) (0.8 mL, 1.6 mmol) was added and stirring was continued. After 115 hours, the reaction was treated with HCl (4 N in dioxane) to pH 5 on pH test strips. The mixture was then partitioned between water and EtOAc. The layers were separated and the aqueous layer was lyophilized to provide a light yellow solid. HPLC purification of preparative RP (YMC-ODS 5 μ column 250 x 30. Conditions: flow rate 30 mL/minute; 40 minute gradient of 30-100% B (Solvent A = 95:5 H2O/MeCN with 0.05% TFA. Solvent B = 5:95 H2O/MeCN with 0.05% TFA) provided the TFA salt of 2-(4-(6-fluoroquinolin-4-yl)piperazin-1 acid -yl)pentanoic acid as a light yellow solid (160.0 mg; 58% production). MS (ES): m/z = 332 [M+H]+. tR = 0.46 minutes (Method A). 1H NMR (400 MHz, DMSO-d6) δ 8.77 (d,J=6.4 Hz, 1H), 8.10 (dd, J=10.1, 5.2 Hz, 1H), 7.93 - 7.86 (m, 2H), 7.28 (d, J=6.5Hz, 1H), 3.84 - 3.73 (m, 4H), 3.72 - 3.51 (m, 1H) , 3.38 - 2.99 (m, 4H), 1.86 - 1.68 (m, 2H), 1.46 - 1.33 (m, 2H), 0.94 (t, J=7, 3 Hz, 3H) Example 244: (±)-N-(4-Chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)pentanamide

[00535] Mixing the salt of (±)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)pentanoic acid (244D, 32.0 mg, 0.10 mmol) in anni - droso (1.5 mL), in a sealable bottle, PyBOP (50.3 mg, 0.10 mmol) was added followed by DIPEA (0.06 mL, 0.34 mmol). The resulting mixture was stirred at room temperature for 10 minutes before 4-chloroaniline (14.8 mg, 0.12 mmol) was added. The vial was sealed and the reaction was stirred at room temperature for 63 hours, before being diluted with DMF, passed through a syringe filter, then purified by preparative HPLC/MS to provide the title compound as a racemate (15.9 mg; 37% production). MS (ES): m/z = 441 [M+H]+. tR = 2.24 minutes (Method B). 1H NMR (500 MHz, DMSO-d6) δ 10.15 (s, 1H), 8.66 (d, J=4.8 Hz, 1H), 8.08 - 7.95 (m, 1H), 7 .68 (d, J=8.7 Hz, 2H), 7.64 - 7.53 (m, 2H), 7.37 (d, J=8.7 Hz, 2H), 7.02 (d, J=4.8 Hz, 1H), 3.57 - 3.43 (m, 1H), 3.35 - 3.08 (m, 4H), 2.94 - 2.78 (m, 4H), 1 .81 - 1.58 (m, 2H), 1.40 - 1.24 (m, 2H), 0.92 (t, J=7.3 Hz, 3H).Example 245 N-(4-Chlorophenyl) -2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)pentanamide (Enantiomer 1, absolute stereochemistry not determined) and Example 246 N-(4-Chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)pentanamide (Enantiomer 2, absolute stereochemistry not determined)

[00536] Racemic example 244 (15.9 mg) was purified by chiral SFC (82/18 CO2/MeOH with 0.1% DEA mobile phase, Chiral OJ column 25 X 3 cm, 5 μm, 85 ml/min, detector wavelength = 220 nm). Concentration of the appropriate fractions (previous elution) provided Example 245 (6.7 mg) assigned as N-(4-chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)pentanamide ( Enantiomer 1). MS (ES): m/z = 441 [M+H]+. Tr = 2.21 minutes (Method B). 1H NMR (500 MHz, DMSO-d6): superimposed over racemate NMR. Concentration of the last elution fractions provided Example 246 (6.8 mg) assigned as N-(4-chlorophenyl)-2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)pentanamide (Enantiomer 2 ). MS (ES): m/z = 441 [M+H]+. Tr = 2.21 minutes (Method B). 1H NMR (500 MHz, DMSO-d6): superimposed over racemate NMR.Example 247 (±)-2-(4-(6-Fluoroquinolin-4-yl)piperazin-1-yl)-N-(1-methylcyclohexyl )pentanamide

[00537] Example 247 (17.5 mg; 42% production) was prepared following a procedure analogous to that for the synthesis of Example 244, except that 1-methylcyclohexanamine, HCl (17.3 mg, 0.12 mmol) was used instead of 4-chloroaniline. MS (ES): m/z = 427 [M+H]+. Tr = 2.23 minutes (Method B). 1H NMR (500 MHz, DMSO-d6) δ 8.64 (d, J=4.8 Hz, 1H), 8.01 (dd, J=9.0, 5.7 Hz, 1H), 7.68 - 7.52 (m, 2H), 7.35 - 7.23 (m, 1H), 7.03 (d, J=4.9 Hz, 1H), 3.19 - 3.09 (m, 4H ), 2.90 - 2.77 (m, 3H), 2.56 - 2.51 (m, 2H), 2.11 - 2.00 (m, 2H), 1.70 - 1.58 (m , 1H), 1.53 - 1.35 (m, 6H), 1.30 - 1.19 (m, 8H), 0.88 (t, J=7.2 Hz, 3H). Example 248 2-(4-(6-Fluoroquinolin-4-yl)piperazin-1-yl)-N-(1-methylcyclohexyl)pentanamide (Enantiomer 1, absolute stereochemistry not determined) and Example 249 2-(4-(6-Fluoroquinolin-4-yl)piperazin-1-yl)-N-(1-methylcyclohexyl)pentanamide (Enantiomer 2, absolute stereochemistry not determined)

[00538] Racemic example 247 (16.7 mg) was purified by chiral SFC (80/20 CO2/MeOH with 0.1% DEA mobile phase, Chiral AD column 25 X 3 cm, 5 μm, 85 ml/min, detector wavelength = 220 nm). Concentration of the appropriate fractions (previous elution) provided Example 248 (7.4 mg) assigned as 2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)-N-(1-methylcyclohexyl)pentanamide ( Enantiomer 1). MS (ES): m/z = 427 [M+H]+. Tr = 2.28 minutes (Method B). 1H NMR (500 MHz, DMSO-d6): superimposed over racemate NMR. Concentration of the last elution fractions provided Example 249 (6.5 mg) assigned as 2-(4-(6-fluoroquinolin-4-yl)piperazin-1-yl)-N-(1-methylcyclohexyl)pentanamide (Enantiomer 2 ). MS (ES): m/z = 427 [M+H]+. Tr = 2.28 minutes (Method B). 1H NMR (500 MHz, DMSO-d6): superimposed over racemate NMR.Example 250 (+/-)-Cys and trans-N-(4-chlorophenyl)-2-(4-(3-methyl-3H-imidazo [4,5-b]pyridin-7-yl)cyclohexyl)propanamide Preparation 250A. 7-chloro-3-methyl-3H-imidazo[4,5-b]pyridine

[00539] To a solution of 7-chloro-3H-imidazo[4,5-b]pyridine, formic acid salt (0.2 g, 0.820 mmol) in DMSO (4.10 ml) was added cesium carbonate ( 0.534 g, 1.639 mmol) and iodomethane (0.054 ml, 0.860 mmol). The reaction is stirred at room temperature for 28 h, then quenched with H2O and extracted with EtOAc (5X). The organics combined, dried over Na2SO4, filtered, and concentrated to provide an orange liquid, which was also dried under high vacuum overnight. TLC and LC-MS showed ~1:1 product/SM. The raw material was dissolved in a minimum amount of CH2Cl2 and subjected to chromatography. Purification of the crude material by silica gel chromatography using an ISCO machine (40 g column, 40 mL/min, 0-20% MeOH in CH2Cl2 for 24 min, tr = 17 min) provided the title compound (0.0729 g, 0.344 mmol, 42.0% yield) as a white solid and 2.8:1 mixture of isomers. ESI MS (M+H)+ = 212.0. HPLC peak tr = 0.55 minutes. HPLC Conditions: Method A.Preparation 250B. Ethyl 2-(4-(3-methyl-3H-imidazo[4,5-b]pyridin-7-yl)cyclohex-3-en-1-yl)propanoate

[00540] Preparation mixture 250A (0.0729 g, 0.435 mmol), 2- (4-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)cyclohex-3- ethyl en-1-yl)propanoate (0.138 g, 0.448 mmol), Na2CO3 (0.184 g, 1.740 mmol), and Pd(Ph3P)4 (0.025 g, 0.022 mmol) in dioxane (3.5 mL) and water ( 0.5 mL) was heated at 100 °C overnight. The reaction was quenched with water and diluted with EtOAc. The layers were separated. The aqueous phase was extracted with EtOAc (3X). The organics were combined, dried over Na2SO4, filtered, and concentrated to provide a brown residue. Purification of the crude material by silica gel chromatography using an ISCO machine (40g column, 40 mL/min, 0-20% MeOH in CH2Cl2 for 25 minutes, tr = 12, 16 min) provided the title compound (79 mg , 0.253 mmol, 58% production) as a colorless residue and its regioisomer (21 mg, 0.067 mmol, 15.41% production) as a colorless residue. ESI MS (M+H)+ = 314.3. HPLC peak tr = 0.75 minutes. HPLC Conditions: Method A.Preparation 250C. Ethyl 2-(4-(3-methyl-3H-imidazo[4,5-b]pyridin-7-yl)cyclohexyl)propanoate

[00541] To a solution of Preparation 250B (0.0792 g, 0.253 mmol) in MeOH (1.264 ml) was added ammonium formate (0.080 g, 1.264 mmol) followed by Pd/C (7.26 mg, 0.068 mmol) . The reaction was heated at 70 °C for 1 h. The reaction was filtered through CELITE® and the filter mass washed with CH2Cl2. The filtrate was concentrated. The raw material was taken up in EtOAc and washed with a saturated aqueous solution of NaHCO3 (2X). The organic phase was dried over Na2SO4, filtered, and concentrated to provide the title compound (59.7 mg, 0.180 mmol, 71% yield) as a green residue. ESI MS (M+H)+ = 316.2. HPLC peak tr = 0.72 minutes. HPLC conditions: Method A.

[00542] Example 250: (+/-)-Cis and trans-N-(4-chlorophenyl)-2-(4-(3-methyl- 3H-imidazo[4,5-b]pyridin-7-yl) cyclohexyl)propanamide

[00543] To a solution of 4-chloroaniline (0.097 g, 0.757 mmol) in THF (0.4 mL) at 0 °C was added a solution of isopropylmagnesium chloride (0.379 mL, 0.757 mmol). The resulting solution was warmed to room temperature and stirred for 5 minutes, then Preparation 250C (0.0597 g, 0.189 mmol) in THF (0.6 mL) was added dropwise. The reaction was heated at 70 °C for 2.5 hours, then allowed to cool to room temperature. The reaction was quenched with a saturated aqueous solution of NH4Cl and diluted with EtO-Ac. The layers were separated. The aqueous phase was extracted with EtOAc (3X). The combined organic phases were dried over Na2SO4, filtered, and concentrated to provide a residue. The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 30 to 70% B for 20 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to provide the title compound as a mixture of 4 isomers (26.6 mg, 35%). ESI MS (M+H)+ = 397.3. HPLC peak tr = 1.775 minutes and 1.793 minutes. Purity = 99%. HPLC Conditions: Method B.Example 251 N-(4-chlorophenyl)-2-(4-(3-methyl-3H-imidazo[4,5-b]pyridin-7-yl)cyclohexyl)propanamide, absolute stereochemistry and relative not confirmed

[00544] Approximately 25.4 mg of diastereomeric and racemic Example 250 were resolved. The isomeric mixture was purified by preparative SFC with the following conditions: Column: Chiral OZ-H, 25 x 3 cm ID, 5 μm particles; Mobile phase A: 82/18 CO2/MeOH with 0.1% DEA; Detector wavelength: 220 nm; Flow: 100 mL/min. The fractions ("Peak-1" tr = 11.910 min, "Peak-2" tr = 15.648 min, "Peak-3" tr = 16.927 min, "Peak-4" tr = 19.403; analytical conditions: Column: Chiral OZ- H, 250 x 4.6 mm ID, 5 μm particles; Mobile phase A: 80/20 CO2/MeOH with 0.1% DEA; Flow: 2.0 mL/min) were collected in MeOH with 0.1 % DEA. The stereoisomeric purity of each fraction was estimated to be greater than 99% based on prep-SFC chromatograms. Each diastereomer or enantiomer was also purified using preparative LC/MS:

[00545] Example 250a, first elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 30 to 70% B for 20 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation. The material was also purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 35-60% B for 25 minutes, then a 2-minute hold at 60% B; Flow: 20 mL/min. Fractions containing the desired product were combined and dried by centrifugal evaporation to give isomer 1 (3.1 mg, 4%). ESI MS (M+H)+ = 397.3. HPLC peak tr = 1,840 minutes. Purity = 95%. HPLC conditions: Method B. Absolute stereochemistry not determined.

[00546] Example 250b, second elution isomer: The raw material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 30 to 70% B for 20 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing the desired product were combined and dried by centrifugal evaporation to provide Isomer 2 (3.0 mg, 4%). ESI MS (M+H)+ = 397.3. HPLC peak tr = 1.804 minutes. Purity = 93%. HPLC conditions: Method B. Absolute stereochemistry not determined.

[00547] Example 250c, third elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 30 to 70% B for 20 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. Fractions containing the desired product were combined and dried by centrifugal evaporation to provide Isomer 3 (3.2 mg, 4%). ESI MS (M+H)+ = 397.3. HPLC peak tr = 1.806 minutes. Purity = 96%. HPLC conditions: Method B. Absolute stereochemistry not determined.

[00548] Example 250d, fourth elution isomer: The crude material was purified using preparative LC/MS with the following conditions: Column: XBridge C18, 19 x 200 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile: water with 10 mM ammonium acetate; Gradient: 30 to 70% B for 20 minutes, then a 5-minute hold at 100% B; Flow: 20 mL/min. The fractions containing the desired product were combined and dried by centrifugal evaporation to provide Isomer 4 (3.0 mg, 4%). ESI MS (M+H)+ = 397.2. HPLC peak tr = 1.841 minutes. Purity = 94%. HPLC conditions: Method B. Absolute stereochemistry not determined. BIOLOGICAL EXAMPLES Example 251

[00549] Assessment of inhibitory activity in indoleamine 2,3-dioxygenase (IDO) assay based on HeLa cells

[00550] HeLa cells (ATCC® CCL-2) were obtained from ATCC and cultured in Dulbecco's Modified Eagle Medium supplemented with 4.5 g/L glucose, 4.5 g/L L-glutamine and 4.5 g /L sodium pyruvate (#10-013-CV, Corning), 2mM L-alanyl-L-glutamine dipeptide (#35050-061, Gibco), 100U/mL penicillin, 100μg/mL streptomi- cin (#SV30010, Hyclone) and 10% fetal serum (#SH30071.03 Hyclone). Cells were maintained in a humidified incubator at 37 °C in 5% CO2.

[00551] IDO activity was measured as a function of kynurenine production as follows: HeLa cells were seeded in a 96-well culture plate at a density of 5,000 cells/wells and allowed to equilibrate overnight. After 24 hours, media were aspirated and replaced with IFNY media (#285-IF/CF, R&D Systems) at a final concentration of 25 ng/mL. A serial dilution of each test compound was added to the cells in a total volume of 200 μL of culture medium. After another 48 incubation, 170 μL of supernatants were transferred to each well of a new 96-well plate. 12.1 μL of 6.1N trichloroacetic acid (#T0699, Sigma-Aldrich) was added to each well and mixed, followed by incubation at 65°C for 20 minutes to hydrolyze N-formylkynurenine, the product of indoleamine 2,3 - dioxygenase, into kynurenine. The reaction mixture was then centrifuged for 10 minutes at 500xg to settle the precipitate. 100 μL of the supernatant was transferred to each well to a new 96-well plate 96. 100 μL of 2% (w/v) p-dimethylaminobenzaldehyde (#15647-7, Sigma-Aldrich) in acetic acid (#A6283, Sigma - Aldrich) were added to each well, mixed and incubated at room temperature for 20 minutes. Kynurenine concentrations were determined by measuring absorbance at 480nm and calibrating against an L-kynurenine standard curve (#K8625, Sigma-Aldrich) using a SPECTRAMAX® M2e plate reader (Molecular Devices). The percent activity at each inhibitor concentration was determined and IC50 values measured using nonlinear regression.

[00552] Activity for the compounds described here is provided in Figures 1A-1P, where potency levels are provided as follows: (Potency: IDO IC50: A < 0.1 μM; B < 1 μM; C < 10 μM) Example 252

[00553] 1HEK293 cells were transfected with a pCDNA-based mammalian expression vector harboring human IDO1 cDNA (NM 002164.2) by electroporation. They were cultured in medium (DMEM with 10% FBS) containing 1 mg/ml G418 for two weeks. HEK293 cell clones that stably expressed human IDO1 protein were selected and expanded for IDO inhibition assay.

[00554] Human IDO1/HEK293 cells were seeded at 10,000 cells per 50μL per well with RPMI/phenol red free media containing 10% FBS in a 384-well clear base, black wall tissue culture plate (Matrix Technologies LLC) 100 nL of a certain concentration of compound was then added to each well using ECHO liquid handling systems. The cells were incubated for 20 hours in an incubator at 37 °C with 5% CO2.

[00555] Compound treatments were stopped by adding Trichloroacetic Acid (Sigma-Aldrich) to a final concentration of 0.2%. The cell plate was also incubated at 50°C for 30 minutes. Equal volume of supernatant (20μL) and 0.2% (w/v) Ehrlich reagent (4-dimethylaminobenzaldehyde, Sigma-Aldrich) in glacial acetic acid were mixed in a new clear-base 384-well plate. This plate was then incubated at room temperature for 30 minutes. Absorbance at 490 nm was measured on an Envision plate reader.

[00556] Compound IC50 values were calculated using counts of 500 nM of a reference standard treatment as one hundred percent inhibition, and counts of no compound treatment but DMSO as zero percent inhibition.

[00557] Compounds with an IC50 greater than 250 nM are shown with (C), compounds with an IC50 less than 250 nM are shown with (B) and those with an IC50 less than 50 nM are shown with (A) in Table X, below. Table X Biological activity for examples tested in the biological assay described in Example 252.

[00558] Particular embodiments of this invention are described here, including the best way known to the inventors to carry out the invention. Upon reading the foregoing description, variations of the described embodiments may become apparent to individuals working in the art, and it is expected that those skilled in the art may employ such variations when appropriate. Consequently, it is intended that the invention be practiced otherwise than as specifically described herein, and that the invention includes all modifications and equivalents of the subject matter reported in the claims attached to this document as permitted by applicable law. Furthermore, any combination of the above-described elements, in all possible variations thereof, is encompassed by the invention, unless otherwise indicated herein or otherwise clearly contradicted by the context.

[00559] All publications, patent applications, accession numbers, and other references cited in this specification are incorporated herein by reference, as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference.

Claims (13)

1. Compound, characterized by the fact that it is (R)-N-(4-chlorophenyl)-2-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)propanamide or a pharmaceutically acceptable salt thereof. 2. Compound according to claim 1, characterized in that it is (R)-N-(4-chlorophenyl)-2-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl) propanamide 3. Compound according to claim 1, characterized by the fact that it is the pharmaceutically acceptable salt of the same (R)-N-(4-chlorophenyl)-2-(cis-4-(6-fluoroquinolin-4-yl) cyclohexyl)propanamide. 4. Pharmaceutical composition, characterized by the fact that it comprises (R)-N-(4-chlorophenyl)-2-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)propanamide or a pharmaceutically acceptable salt thereof as defined in claim 1 and a pharmaceutically acceptable excipient. 5. Pharmaceutical composition according to claim 4, characterized by the fact that it comprises (R)-N-(4-chlorophenyl)-2-(cis-4-(6-fluoroquinolin-4-yl)cyclohexyl)propanamide 6. Pharmaceutical composition according to claim 4, characterized by the fact that it comprises a pharmaceutically acceptable salt of (R)-N-(4-chlorophenyl)-2-(cis-4-(6-fluoroquinolin-4-yl) cyclohexyl)propanamide. 7. Use of a compound as defined in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that it is in the preparation of a medicament for the treatment of cancer. 8. Use according to claim 7, characterized in that said cancer is cancer of the prostate, colon, rectum, pancreas, cervix, stomach, endometrium, brain, liver, bladder, ovary, testicle, head, neck, skin (including melanoma and basal cell carcinoma), mesothelial lining, white blood cells (including lymphoma and leukemia), esophagus, breast, muscle, connective tissue, lung (including small cell carcinoma and non-small cell carcinoma), adrenal gland, thyroid, kidney, or bone; or is glioblastoma, mesothelioma, renal cell carcinoma, gastric carcinoma, sarcoma (including Kaposi's sarcoma), choriocarcinoma, cutaneous basal cell carcinoma, or testicular seminoma; or optionally wherein said cancer is selected from the group consisting of melanoma, colon cancer, pancreatic cancer, breast cancer, prostate cancer, lung cancer, cervical cancer, leukemia, sarcoma, kidney cancer, a brain tumor, lymphoma, ovarian cancer, and Kaposi's sarcoma. 9. Combination, characterized by the fact that it comprises a compound as defined in claim 1, and at least one additional therapeutic agent. 10. Combination according to claim 9, characterized by the fact that the at least one additional therapeutic agent is a chemotherapeutic agent, an immunomodulating and/or inflammation agent, an antihypercholesterolemia agent, or an anti-infectious agent . 11. Combination according to claim 9, characterized by the fact that the at least one additional therapeutic agent is an immunological checkpoint inhibitor. 12. Use of a compound as defined in claim 1, or a pharmaceutically acceptable salt thereof, characterized in that it is in the preparation of a medicament for the treatment of cancer in combination with an immunological checkpoint inhibitor. 13. Use according to claim 12, characterized by the fact that said immune checkpoint inhibitor is selected from the group consisting of ipilimumab, nivolumab and pembroluzimab.