Inhibition of human integrin α4β7

Abstract

To provide a pharmaceutical composition suitable for the inhibition of α4β7 integrin.SOLUTION: A pharmaceutical composition includes a specific compound with a partial structure of 2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl.SELECTED DRAWING: Figure 1-1

Description

Technical Field

[0001] Related Applications This application claims the benefit of priority to PCT International Application No. PCT / US20 / 55986, filed October 16, 2020.

[0002] Disclosed are novel compounds and related methods useful for the inhibition of α4β7 integrin. The compounds and methods disclosed herein are applicable to the development of medicaments for the treatment of α4β7 integrin-mediated conditions such as inflammatory bowel disease (IBD), ulcerative colitis (UC) and Crohn's disease (CD).

Background Art

[0003] Integrins are non-covalently associated α / β heterodimeric cell surface receptors that are involved in a number of cellular processes. Differential expression of integrins can regulate the adhesive properties of cells, allowing different leukocyte populations to be mobilized to specific organs in response to different inflammatory signals. α4 integrins, including α4β7, play a role in lymphocyte trafficking throughout the gastrointestinal tract. They are expressed on most leukocytes, including B and T lymphocytes, where they mediate cell adhesion through selective binding to their primary ligand, mucosal addressin cell adhesion molecule (MAdCAM). Memory T lymphocytes expressing α4β7 integrin preferentially migrate into the gastrointestinal tract through strong adhesion to mucosal vascular addressin cell adhesion molecule 1 (MAdCAM-1).

[0004] Inhibitors of specific integrin-ligand interactions have been used for the treatment of various diseases. For example, monoclonal antibodies that exhibit high binding affinity for α4β7 have shown therapeutic benefits for gastrointestinal autoinflammatory / autoimmune diseases, such as Crohn's disease and ulcerative colitis. However, these treatments also have certain undesirable characteristics for patients. Monoclonal antibody α4β7 integrin inhibitors are administered by parenteral administration, have a long half-life that cannot rapidly modify exposure, and reduced activity due to anti-drug antibody formation. Monoclonal antibody treatments can be difficult to manufacture compared to small molecule treatments. In addition, some treatments that inhibit α4β7 also interfere with the α4β1 integrin-ligand interaction, thereby causing dangerous side effects to patients. Activity at α4β1 integrin is associated with the emergence of progressive multifocal leukoencephalopathy (PML), a life-threatening and progressive brain infection in immunosuppressed patients.

[0005] As an important addition to therapeutic medical supplies for α4β7 integrin-mediated conditions, such as inflammatory bowel disease (IBD), ulcerative colitis (UC) and Crohn's disease (CD), there remains a medical need for effective and safe oral α4β7 integrin inhibitors with improved pharmaceutical properties.

Summary of the Invention

Means for Solving the Problems

[0006] The present invention relates to compounds that antagonize α4β7 integrin, and methods for preparing and isolating the compounds. In addition, the small molecule compounds disclosed herein exhibit good pharmaceutical properties, including oral bioavailability, ADME (absorption, distribution, metabolism and excretion), pharmacokinetics, CYP inhibition, and / or other safety profiles useful for obtaining therapeutic efficacy while minimizing undesired characteristics.

Brief Description of the Drawings

[0007] [Figure 1-1]This table summarizes the in vitro inhibition of α4β7 integrin by illustrative compounds (i.e., data obtained from the fluorescence polarization assay in Example 4 and the ligand binding assay in Example 5). [Figure 1-2] This table summarizes the in vitro inhibition of α4β7 integrin by illustrative compounds (i.e., data obtained from the fluorescence polarization assay in Example 4 and the ligand binding assay in Example 5). [Figure 1-3] This table summarizes the in vitro inhibition of α4β7 integrin by illustrative compounds (i.e., data obtained from the fluorescence polarization assay in Example 4 and the ligand binding assay in Example 5). [Figure 1-4] This table summarizes the in vitro inhibition of α4β7 integrin by illustrative compounds (i.e., data obtained from the fluorescence polarization assay in Example 4 and the ligand binding assay in Example 5). [Modes for carrying out the invention]

[0008] In certain embodiments, the present invention relates to compounds that antagonize α4β7 integrin. These compounds are useful in treating diseases that can be treated by inhibiting α4β7 integrin (e.g., Crohn's disease (CD) and ulcerative colitis (UC)).

[0009] definition For convenience, prior to any further description of the invention, certain terms used in this specification, the examples, and the appended claims are set forth herein. These definitions should be read in light of the remainder of this disclosure and understood as those skilled in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art.

[0010] To make the present invention easier to understand, certain terms and phrases are defined below and throughout this specification.

[0011] The articles "a" and "an" are used herein to refer to one or more than one (i.e., at least one) grammatical object of the article. For example, "an element" means one or more elements.

[0012] The phrase “and / or” in this specification and claims should be understood, as used herein, to mean “either or both” of elements that are thus coordinately connected, i.e., elements that are sometimes seen as connective and sometimes as disjunctive. Multiple elements listed with “and / or” should be interpreted in the same manner, i.e., “one or more” of elements that are thus coordinately connected. Other elements may exist, whether related or unrelated to the elements specifically identified by the “and / or” clause, as they may be. Thus, as a non-restrictive example, a reference to “A and / or B,” when used in conjunction with open-ended language, for example, “comprising,” may refer to, in one embodiment, only A (including, if applicable, elements other than B), in another embodiment, only B (including, if applicable, elements other than A), and in yet another embodiment, both A and B (including, if applicable, other elements), and so on.

[0013] Where used herein and in the claims, “or” should be understood to have the same meaning as “and / or” as defined above. For example, when separating items in a list, “or” or “and / or” should be interpreted as inclusive, that is, including more than one, not just one, of the multiple or elements of the list, and, if applicable, additional unlisted items. Only terms clearly indicated to the contrary, such as “exactly one of” or “exactly one of,” or, where used in a claim, “consisting of,” refer to the inclusion of exactly one of the multiple or elements of the list. In general, the term “or,” where used herein, should be interpreted as indicating only exclusive substitutes (i.e., “one or the other, but not both”) when preceded by terms of exclusivity, such as “either,” “one of,” “exactly one of,” or “exactly one of.” “Must be from” has its usual meaning where it is used in the field of patent law.

[0014] As used herein and in the claims, the phrase “at least one” should be understood to mean, in reference to a list of one or more elements, at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed in the list of elements, and not excluding any combination of elements in the list of elements. This definition also allows for elements to exist other than those specifically identified in the list of elements to which the phrase “at least one” refers, whether related or unrelated to the elements that are specifically identified. Therefore, as a non-restrictive example, "at least one of A and B" (or equally "at least one of A or B", or equally "at least one of A and / or B") may refer to, in one embodiment, at least one A, including more than one, in which B is absent (and optionally including elements other than B); in another embodiment, at least one B, including more than one, in which A is absent (and optionally including elements other than A); and in yet another embodiment, at least one A, including more than one, and at least one B, including more than one (and optionally including other elements).

[0015] Furthermore, unless otherwise clearly indicated, in any method claimed herein that includes more than one step or act, the order of the steps or acts of the method should be understood not to be limited to the order in which the steps or acts of the method are enumerated.

[0016] In the claims, and similarly in this specification, all transitional phrases, such as “comprising,” “including,” “possessing,” “having,” “containing,” “accompanying,” “holding,” and “composed of,” should be understood to be open-ended, that is, to mean including but not limiting. Only the transitional phrases “consisting of” and “essentially from” are closed or semi-closed transitional phrases, as described in the United States Patent and Trademark Office Prosecution Procedure Manual 2111.03.

[0017] Certain compounds contained in the compositions of the present invention may exist in special geometric or stereoisomeric forms. In addition, the polymers of the present invention may also be optically active. The present invention intends that all such compounds, including cis and trans isomers, R and S enantiomers, diastereomers, (D) isomers, (L) isomers, racemic mixtures thereof, and other mixtures, fall within the scope of the invention. Additional chiral carbon atoms may be present in substituents, for example, alkyl groups. All such isomers, as well as mixtures thereof, are intended to be included in this invention.

[0018] For example, if a specific enantiomer of the compound of the present invention is desired, it can be prepared by asymmetric synthesis or by induction using a chiral auxiliary agent, where the resulting diastereomer mixture is separated and the auxiliary groups are cleaved to provide the pure desired enantiomer. Alternatively, if the molecule contains a basic functional group, e.g., amino, or an acidic functional group, e.g., carboxyl, a diastereomer salt is formed with a suitable optically active acid or base, followed by the separation of the thus formed diastereomer by fractional crystallization or chromatographic means known in the art, and subsequent recovery of the pure enantiomer.

[0019] The structures illustrated herein are to be interpreted as containing different compounds only in the presence of one or more isotopically enriched atoms. For example, hydrogen substitution with deuterium or tritium, or 13 C or 14 Compounds produced by carbon substitution with carbon-enriched carbon fall within the scope of this invention.

[0020] When used herein, terms such as "α4β7," "a4B7," "a4b7," "alpha-4beta-7," and "alpha-4beta7" all refer to α4β7.

[0021] The phrases “pharmaceutically acceptable excipient” or “pharmaceutically acceptable carrier,” as used herein, mean a pharmaceutically acceptable material, composition or vehicle involved in holding or transporting the chemical substance of interest from one organ or part of the body to another, such as a liquid or solid filler, diluent, excipient, solvent or encapsulating material. Each carrier must be “acceptable” in the sense that it is compatible with the other components of the formulation, is not harmful to the patient, and is substantially nonpyrogenic. Some examples of materials that can act as pharmaceutically acceptable carriers include: (1) sugars, e.g., lactose, glucose and sucrose; (2) starches, e.g., corn starch and potato starch; (3) cellulose and its derivatives, e.g., sodium carboxymethylcellulose, ethylcellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, e.g., cocoa butter; (9) oils, e.g., peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and (10) Soybean oil, (11) Glycol, e.g., propylene glycol, (12) Polyol, e.g., glycerin, sorbitol, mannitol, and polyethylene glycol, (13) Ester, e.g., ethyl oleate and ethyl laurate, (14) Agar, (15) Buffering agent, e.g., magnesium hydroxide and aluminum hydroxide, (16) Alginic acid, (17) Pyrogen-free water, (18) Isotonic saline, (19) Ringer's solution, (16) Ethyl alcohol, (20) Phosphate buffer solution, and (21) Other non-toxic, suitable substances used in pharmaceutical formulations. In certain embodiments, the pharmaceutical compositions of the present invention are non-pyrogenic, i.e., they do not induce a significant rise in temperature when administered to a patient.

[0022] The term "pharmaceutically acceptable salts" refers to relatively non-toxic inorganic and organic acid addition salts of a compound. These salts can be prepared in situ during the final isolation and purification of the compound, or by reacting the purified compound separately with a suitable organic or inorganic acid in its free base form, and then isolating the salt thus formed. Representative salts include hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, naphthylate, mesylate, glucoheptonate, lactobionate, and laurylsulfonate. (See, for example, Berge et al. (1977) "Pharmaceutical Salts," J. Pharm. Sci. 66:1-19.)

[0023] In other cases, compounds useful in the methods of the present invention may contain one or more acidic functional groups and thus be able to form pharmaceutically acceptable salts with pharmaceutically acceptable bases. In these examples, the term "pharmaceutically acceptable salt" refers to a relatively non-toxic inorganic or organic base addition salt of the compound. These salts can also be prepared in situ during the final isolation and purification of the compound, or by reacting the purified compound in its free acid form separately with a suitable base, such as a hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary, or tertiary amine. Typical alkali salts or alkaline earth salts include lithium salts, sodium salts, potassium salts, calcium salts, magnesium salts, and aluminum salts. Typical organic amines useful for forming base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, and piperazine (see, e.g., Berge et al., above).

[0024] The “therapeutic effective dose” (or “effective dose”) of a compound for use in treatment refers to the amount of the compound in a preparation that, when administered (to mammals, preferably humans) as part of a desired dosage regime, reduces symptoms, alleviates a condition, or delays the onset of a disease state, for any disorder or condition to be treated or for cosmetic purposes, in accordance with clinically acceptable standards, at a reasonable benefit / risk ratio applicable to any medical treatment.

[0025] The terms “preventive or therapeutic” treatments are technically recognized and include the administration of one or more of the composition in question to a host. If it is administered before the clinical manifestation of an undesirable condition (e.g., disease or other undesirable condition in the host animal), the treatment is preventive (i.e., it protects the host from developing the undesirable condition); on the other hand, if it is administered after the manifestation of the undesirable condition, the treatment is therapeutic (i.e., it is intended to alleviate, relieve, or stabilize the existing undesirable condition or its side effects).

[0026] The term "patient" refers to a mammal requiring special treatment. In certain embodiments, the patient is a primate, dog, cat, or horse. In certain embodiments, the patient is a human.

[0027] As used herein, the term "prodrug" encompasses compounds that are converted into therapeutically active agents under physiological conditions. A common method for producing prodrugs involves including a selective moiety that reveals a desired molecule upon hydrolysis under physiological conditions. In other embodiments, prodrugs are converted by the enzymatic activity of a host animal.

[0028] For the purposes of this invention, the chemical elements are identified according to the Periodic Table of Elements, CAS version, Handbook of Chemistry and Physics, 67th edition, 1986-87, inside front cover.

[0029] Exemplary compounds In a particular embodiment, the present invention relates to a compound selected from the group consisting of:

[0030] [ka] JPEG0007869261000002.jpg215149JPEG0007869261000003.jpg140157

[0031] In certain embodiments, the present invention relates to one of the compounds described above, wherein the compound is in the form of a pharmaceutically acceptable salt.

[0032] In a particular embodiment, the present invention relates to a compound selected from the group consisting of:

[0033] [Table 1] JPEG0007869261000005.jpg210150JPEG0007869261000006.jpg200150JPEG0007869261000007.jpg147151

[0034] Exemplary pharmaceutical compositions The compounds disclosed herein can be formulated in a variety of pharmaceutical compositions. The compounds disclosed herein, as well as their pharmaceutically acceptable salts, may be pharmaceutically active ingredients (APIs) that are combined with one or more other components to form a drug substance pharmaceutical composition. A drug substance (DS) pharmaceutical composition may include an API (i.e., the compounds disclosed herein or their pharmaceutically acceptable salts) and one or more pharmaceutically acceptable carriers, diluents, and / or excipients. The carriers, diluents, or excipients may be selected to be compatible with the other components of the formulation and to be appropriately safe and effective for the intended treatment. A desired weight concentration of a compound as a pharmaceutically active ingredient (API) can be combined with other inactive components to form a drug substance (DS) in a formulation batch. Pharmaceutically acceptable compositions can be formulated for administration by an appropriate route, for example, by oral delivery in a unit dosage form (including as capsules or tablets). Such compositions can be prepared by associating a pharmaceutically active ingredient (API) containing the compound of formula (I) with a carrier or excipient.

[0035] In certain embodiments, the present invention provides a pharmaceutical composition formulated for the oral delivery of an α4β7 integrin inhibitor, the composition comprising an α4β7 integrin inhibitor compound as an API and a pharmaceutically acceptable carrier formulated for the oral therapeutic administration of the α4β7 integrin inhibitor compound.

[0036] In certain embodiments, the present invention provides a pharmaceutical composition comprising a compound or a pharmaceutically acceptable salt thereof as a pharmaceutically active ingredient (API).

[0037] In a particular embodiment, the present invention relates to a pharmaceutical composition comprising a compound selected from the group consisting of the following as an active pharmaceutical ingredient (API) or a pharmaceutically acceptable salt thereof:

[0038] [Table 2] JPEG0007869261000009.jpg210151JPEG0007869261000010.jpg201151JPEG0007869261000011.jpg148150

[0039] A pharmaceutically acceptable composition containing the compound of the present invention can be prepared by various procedures. For example, the compound of the present invention can be formulated with a suitable excipient, diluent, or carrier to form tablets or capsules and other suitable dosage forms.

[0040] Pharmaceutical compositions can be provided in unit dose form containing a predetermined amount of API comprising the compound of the present invention per unit dose. Such units may contain a desired amount of the compound or a pharmaceutically acceptable salt thereof, depending on the condition being treated, the route of administration, and the patient's age, weight, and condition. Such unit doses can therefore be administered at desired dose intervals. The concentration of the active compound in a drug composition depends on various applicable parameters and considerations, such as the absorption, inactivation, and excretion rates of the drug, as well as other factors known to those skilled in the art. It should be noted that dose values ​​also vary with the severity of the condition to be alleviated. For any specific target, a particular dose regimen should be adjusted over time according to individual needs and the professional judgment of the person administering or supervising the administration of the composition, and it should be further understood that the concentration ranges described herein are illustrative only and are not intended to limit the scope or practice of the claimed compositions. The active ingredient may be administered at once or divided into a number of smaller doses at varying time intervals.

[0041] In certain embodiments, the mode of administration of the active compound is oral. Oral compositions generally include an inert diluent or food carrier. They can be encapsulated in gelatin capsules or compressed into tablets. For the purpose of oral therapeutic administration, the active compound can be incorporated with excipients and used in the form of tablets, lozenges, or capsules. Pharmaceutically compatible binders and / or adjuvant materials can be included as part of the composition. Pharmaceutical compositions containing the compounds of the present invention formulated for oral delivery can be prepared in unit dosage forms, for example, in capsules, at the desired dose strength of the compound. For oral administration in liquid form, the oral drug components can be combined with any oral, non-toxic, pharmaceutically acceptable inert carrier, such as ethanol, glycerol, or water. For oral administration in tablet or capsule form, the compounds of the present invention can be combined with an oral, non-toxic, pharmaceutically acceptable inert carrier. Other examples of excipients, diluents, and carriers suitable for these formulations include: fillers and bulking agents, e.g., starch and sugar; and binders, e.g., cellulose derivatives. Furthermore, suitable binders, lubricants, disintegrants, and colorants may be incorporated into the mixture as desired or required. Suitable binders include starch, natural sugars, natural or synthetic gums, etc. Lubricants and / or flow enhancers may be used in these formulations.

[0042] Tablets, pills, capsules, lozenges, etc., may contain any of the following components or compounds of similar properties: binders, e.g., microcrystalline cellulose, tragacanth gum, or gelatin; excipients, e.g., starch, or lactose; disintegrants, e.g., alginic acid, Primogel, or corn starch; lubricants, e.g., magnesium stearate or Sterotes; flow enhancers, e.g., colloidal silicon dioxide; sweeteners, e.g., sucrose, or saccharin; or flavoring agents, e.g., peppermint, methyl salicylate, or orange flavoring. If the dosage unit form is a capsule, it may contain a liquid carrier, e.g., fatty oil, in addition to the above-mentioned materials. In addition, the dosage unit form may contain various other materials that modify the physical form of the dosage unit, e.g., sugar coatings, or other enteric coatings.

[0043] The compounds can be administered as components of elixirs, suspensions, syrups, wafers, etc. In addition to the active compound(s), the syrup may contain sucrose or other sweeteners as a sweetener, as well as certain preservatives, dyes and colorants, and flavorings.

[0044] The compounds can be formulated as solutions suitable for parenteral administration, for example, via intramuscular, subcutaneous, or intravenous routes. For example, the compounds of the present invention can be dissolved in a suitable buffer solution. Pharmaceutical compositions containing a desired concentration of the compounds of the present invention can be formulated as injectable drug solutions (e.g., useful in preclinical animal studies).

[0045] Illustrative methods Compounds that inhibit α4β7 are useful in the development of medicines to treat patients with ulcerative colitis and Crohn's disease. Patients with ulcerative colitis (UC) and Crohn's disease (CD) suffer from autoimmune inflammation in the gastrointestinal tract, and for many of these patients, CD4 +Memory T cells drive disease progression and relapses in the gut through their ability to secrete pro-inflammatory effector cytokines, affecting surrounding immune cells and tissues. The progression and relapse of these disease states are thought to involve extravasation of T cells, which leave the bloodstream and invade tissues within the gut, leading to the inflammatory state found in UC and CD via integrin-related mechanisms. Inhibition of α4β7 disrupts this mechanism, thereby preventing T cell localization to tissues and effectively treating and preventing diseases such as UC and CD. T cell homing to the gut requires the surface expression of integrin α4β7 and the chemokine receptor CCR9. CCR9 is utilized by cells to migrate against the gradient of CCL25 expressed in the small intestine, while α4β7 is a ligand-binding anchoring molecule, mucosal addressin cell adhesion molecule 1 (MAdCAM-1). Integrin α4β7 binds to MAdCAM-1 with high affinity, facilitating cell rolling and firm adhesion, followed by extravasation into tissue.

[0046] The pharmaceutical composition may include a compound that inhibits α4β7 integrin on inflammatory cells, thereby inhibiting or preventing the adhesion of these cells to mucosal adresin cell adhesion molecule-1 (MAdCAM-1), and preventing these cells from invading the lamina propria and intestinal lymphoid tissues.

[0047] As described in Example 4, the compounds of the present invention were evaluated using a fluorescence polarization (FP) assay. The FP assay is used to evaluate the potency of the compound on purified protein. The FP assay consists of measuring a headpiece that binds to the purified integrin αβ heterodimer ectodomain or a substitute or truncated ligand. The results of the FP assay for illustrative compounds of the present invention are provided herein.

[0048] The compounds of the present invention were further evaluated using the ligand-binding assay (LBA) described in Example 5 for examining the compound potency of free ligands that bind to receptors expressed on cells. The MAdCAM ligand-binding assay measures the binding of fluorescently labeled MAdCAM-1-Fc to RPMI 8866 cells in the presence of Mn++ using flow cytometry. This assay determines the binding of the compound to the undenatured full-length receptor on the cell surface. One advantage of the MAdCAM ligand-binding assay is its ability to quantify and identify the potent activity of compounds beyond the functional sensitivity limit of the FP assay [approximately 10 nM in Mn]. Ligand-binding assays (LBAs) are used to examine compound potency and the selectivity of free ligands that bind to receptors expressed on cells.

[0049] In certain embodiments, the present invention relates to any one of the previously described methods, wherein the subject is a mammal. In certain embodiments, the present invention relates to any one of the previously described methods, wherein the subject is a human. [Examples]

[0050] The present invention is now generally described and will be more readily understood by referring to the following examples, which are listed solely for the purpose of illustrating certain aspects and embodiments of the invention and are not intended to limit the invention.

[0051] Examples 1-3 describe the synthesis of the compounds of the present invention. The compounds can be prepared as a mixture of diastereomer compounds having a (3S) configuration (i.e., with a stereocenter that is beta relative to the carboxylic acid moiety), and as a mixture of diastereomers with a chiral center covalently bonded to the pyridone ring nitrogen atom of the compound.

[0052] The compound exhibiting greater activity in the fluorescence polarization (FP) assay of Example 4 is shown in a specific diastereomer isomer. Example 4 describes the fluorescence polarization (FP) assay. Example 5 describes the ligand binding (LB) assay. Example 6 describes the cell adhesion (CA) assay.

[0053] [Example 1] General scheme for the synthesis of α4β7 inhibitors β-amino acid synthesis The synthesis of β-amino acids can be achieved using well-known procedures described in the literature, for example, "Enantioselective Synthesis of β-Amino Acids," 2nd edition, editors: Eusebio Juaristi, Vadim A. Soloshonok, first published January 27, 2005, John Wiley & Sons, Inc., Ellman et al., Acc. Chem. Res. 2002, 35, pp. 984-995; Franklin A. Davis and Bang-Chi Chen Chem., Soc. Rev., 1998, 27, pp. 13-18; Jacobsen, MF, Skrydstrup, TJ Org. Chem. 2003, 68, pp. 7122; Tang, TP, Ellman, JAJ Org. Chem. 2002, 67, pp. 7819; and Tang, TP, Ellman, JAJ Org. This is the referenced source, but is not limited to, Chem. 1999, pages 64 and 12.

[0054] Reductive amination

[0055] [ka] Procedure A: A mixture of amine (1 equivalent) and aldehyde (1.2 equivalents) in DCM (1-2 mL / mmol amine) was stirred at room temperature for 30 minutes. Then, NaBH(OAc)3 (1.5 equivalents) was added in small amounts, and the mixture was stirred overnight at room temperature. The solvent was concentrated under vacuum, and the residue was purified by silica gel chromatography to obtain the desired amine. Procedure B: A mixture of aldehyde (1 equivalent) and amine (1.05-2 equivalents) in DCE (3-4 mL / mmol aldehyde) was stirred at room temperature for 10-30 minutes. Then, NaBH(OAc)3 (3-4 equivalents) was added in small amounts, and the mixture was stirred at room temperature for 1-16 hours until LC / MS analysis was completed. The solvent was concentrated under vacuum, and the residue was purified by silica gel chromatography to obtain the desired amine. Procedure C: A mixture of aldehyde (1 equivalent), AcOH (1.2 equivalents), and amine (1.05-2 equivalents) in DCM (2-3 mL / mmol aldehyde) and MeOH (0.5 mL / mmol aldehyde) was stirred at room temperature for 15-30 minutes. Then, NaBH(OAc)3 (2 equivalents) was added in small amounts, and the mixture was stirred at room temperature for 1-16 hours until LC / MS analysis was completed. The solvent was concentrated under vacuum, and the residue was purified by silica gel chromatography to obtain the desired amine.

[0056] Alkylation

[0057] [ka] Procedure A: To a solution of amine (1 equivalent) in MeCN (3-4 mL / mmol amine), mesylate (1.5 equivalents) and K2CO3 (3 equivalents) were added. The reaction mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under vacuum, and the residue was purified by reverse-phase HPLC to obtain the alkylation product.

[0058] [ka] Procedure B: Alkyl bromide (2 equivalents) and K2CO3 (2 equivalents) were added to a solution of amine (1 equivalent) in MeCN (3-4 mL / mmol amine). The reaction mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under vacuum, and the residue was purified by reverse-phase HPLC to obtain the alkylation product.

[0059] Phenol deprotection

[0060] [ka] A mixture of methoxypyridine (1 equivalent) in 44% HBr / AcOH (10 mL / mmol substrate) was heated at 55–75°C for 5–16 hours until LC-MS analysis was complete. The reaction product was concentrated under vacuum, and the residue was purified by reverse-phase HPLC to obtain the phenol product.

[0061] Wittig reaction

[0062] [ka] Procedure A: A mixture of (methoxymethyl)triphenylphosphonium chloride (1.5 equivalents) and t-BuOK (2.5 equivalents) in dioxane (2 mL / mmol phosphonium salt) was stirred at room temperature for 15 minutes. Then, an aldehyde (1 equivalent) in THF (1 mL / mmol aldehyde) was added. The mixture was stirred at room temperature for 2 to 16 hours. The reaction mixture was post-treated (diluted with water, extracted with siRNA, the extracts were combined, dehydrated with Na2SO4, filtered and concentrated), and purified by silica gel chromatography to obtain the enol ether product. Procedure B: A mixture of (methoxymethyl)triphenylphosphonium chloride (1.1 equivalents) and t-BuOK (2.5 equivalents) in THF (4 mL / mmol phosphonium salt) was stirred at 0°C for 1 hour. Then, an aldehyde (1 equivalent) in THF (2 mL / mmol aldehyde) was added. The mixture was stirred at room temperature for 16 hours. The reaction mixture was post-treated (diluted with water, extracted with ELISA, the extracts were combined, dehydrated with Na2SO4, filtered, and concentrated), and purified by silica gel chromatography to obtain the enol ether product.

[0063] From enol ether to aldehyde

[0064] [ka] Procedure A: Enol ether (1 equivalent) was treated with TFA (2 mL / mmol) at room temperature for 4 hours. The solvent was removed under vacuum to obtain the desired aldehyde. Procedure B: Enol ether (1 equivalent) was treated with HCOOH (2 mL / mmol) at 70°C for 2 hours. The solvent was removed under vacuum to obtain the desired aldehyde. Procedure C: To a solution of enol ether (1 equivalent) in DCM (15 mL / mmol enol ether), TFA (2 mL / mmol) and water (0.25 mL / mmol enol ether) were added. The reaction mixture was stirred at 45°C for 18 hours. The reaction mixture was work-treated (reaction stopped with NaHCO3, extracted with DCM, extracts combined, dehydrated with Na2SO4, filtered, and concentrated) to obtain the desired aldehyde.

[0065] Stille reaction

[0066] [ka] To a solution of aryl bromide (1 equivalent) and allyl stanane (1.2 equivalents) in DMF (3 mL / mmol aryl bromide), Pd(PPh3)4 (0.1 equivalent) was added under N2. The reaction mixture was stirred at 100°C for 16 hours. The reaction mixture was concentrated under vacuum, then diluted with SiO2, injected into 20% aqueous KF, and extracted by stirring for 1 hour. The combined organic layer was dehydrated with Na2SO4, filtered and concentrated, and purified by silica gel chromatography to obtain the desired product.

[0067] Alkenes to aldehydes

[0068] [ka] To a solution of alkene (1 equivalent) in THF / H2O (1:1) (10 mL / mmol alkene) at 0°C, K2OsO4-2H2O (0.01 equivalent) was added. The mixture was stirred at 0°C for 5 minutes, then NaIO4 (3 equivalents) was added dropwise to H2O (1 mL / mmol alkene), and the mixture was stirred at 0°C for 1 hour. The mixture was then warmed to room temperature and stirred until completion by LC-MS. The reaction product was work-treated (diluted with water and extracted with siRNA, the combined organic layer was dehydrated with Na2SO4, filtered, and concentrated) to obtain the desired aldehyde.

[0069] From esters to acids

[0070] [ka] One equivalent of the ester was treated with LiOH-H2O (3-5 equivalents) in MeOH (1-3 mL / mmol ester) and water (1-3 mL / mmol ester) at room temperature for 1-5 hours. The reaction mixture was acidified to pH=3 with 1N HCl and concentrated. The residue was purified by preparative HPLC to obtain the desired carboxylic acid product.

[0071] Amine protection

[0072] [ka] A mixture of amine (1 equivalent), DIEA (3 equivalents), and Boc2O (2 equivalents) was stirred in DCM (5 mL / mmol amine) at room temperature for 16 hours until LC-MS analysis was completed. The reaction mixture was work-treated (washed with 0.5 N HCl, saturated NaHCO3, and brine, extracted with DCM, the combined organic layer was dehydrated with Na2SO4, filtered, concentrated), and purified by silica gel chromatography.

[0073] Preparation of arylboranes

[0074] [ka] A mixture of aryl bromide (1 equivalent), B2pin2 (1.2 equivalents), Pd(dppf)Cl2 (0.05 equivalent), and KOAc (3 equivalents) in dioxane (10 mL / mmol aryl bromide) was stirred under N2 at 110°C for 2-5 hours until LC-MS analysis was completed. The reaction product was filtered, concentrated under vacuum, and purified by silica gel chromatography to obtain the desired arylborane.

[0075] Suzuki Coupling "Palladium-Catalyzed Cross-Coupling Reactions of Organoboron Compounds", N. Miyaura, A. Suzuki Chem. Rev.1995, 957, pp. 2457-2483.

[0076] [ka] Procedure A: To a solution of arylborane (1 equivalent) in dioxane (10 mL / mmol arylborane), aryl bromide (1.2 equivalents), Pd(dppf)Cl2 (0.1 equivalent), K2CO3 (2 equivalents), and water (2 mL / mmol) were added. The reaction mixture was stirred under N2 at 110°C for 3 hours. The reaction mixture was work-treated (washed with saline solution, extracted with SiO2, the extracts were combined, dehydrated with Na2SO4, filtered, and concentrated), and purified by silica gel chromatography to obtain the desired biaryl product.

[0077] [ka] Procedure B: To a solution of aryl bromide (1 equivalent) and arylborane (1.1 equivalents) in dioxane (10 mL / mmol aryl bromide), water (2 mL / mmol) and K2CO3 (2 equivalents) in Pd(dppf)Cl2 (0.1 equivalent) were added. The reaction mixture was stirred under N2 at 110°C for 2 hours. The reaction mixture was work-treated (washed with saline solution, extracted with SiO2, the extracts were combined, dehydrated with Na2SO4, filtered, and concentrated), and purified by silica gel chromatography to obtain the desired biaryl product.

[0078] [ka] Procedure C: A mixture of aryl bromide (1 equivalent), arylborane (2.0 equivalents), K2CO3 (3 equivalents), and Pd(dppf)Cl2 (0.05 equivalents) in dioxane (10 mL / mmol aryl bromide) and water (1 mL / mmol) was stirred at 110°C for 2 hours under N2 until LC-MS analysis was completed. The reaction product was work-treated (washed with brine, extracted with SiO2, the extracts were combined, dehydrated with Na2SO4, filtered, and concentrated), and purified by silica gel chromatography to obtain the desired biaryl product.

[0079] Boc deprotection

[0080] [ka] 4 M HCl-dioxane (12 equivalents) was added to DCM (4 mL / mmol amine) containing a Boc-protected amine (1 equivalent). The reaction was stirred for 1-2 hours until completion by LC-MS. The reaction was concentrated under vacuum to obtain the desired amine. t-butylsulfinyl deprotection

[0081] [ka] To a solution of t-butylsulfinylamine (1 equivalent) in DCM (0.5 mL / mmol amine), 4M HCl-dioxane (1.7 equivalents) was added. The reaction was stirred for 0.5 to 1 hour until completion by LC-MS. The reaction was concentrated and purified by preparative HPLC to obtain the desired amine.

[0082] Amide bond formation "Peptide Coupling Reagents, More than a Letter Soup", A. El-Faham, F. Albericio Chem. Rev. 2011, 111, 11, pp. 6557-6602; "Amide bond formation and peptide coupling", CAGN Montalbetti, V. Falque Tetrahedron 2005, 61, pp. 10827-10852.

[0083] [ka] A mixture of amine (1 equivalent), carboxylic acid (1 equivalent), TCFH (2 equivalents), and NMI (4 equivalents) in CH3CN (10 mL / mmol amine) was stirred at room temperature for 1-2 hours until LC-MS analysis was completed. The reaction mixture was concentrated under vacuum and purified by silica gel chromatography to obtain the desired amide product.

[0084] Ester hydrolysis

[0085] [ka] One equivalent of the ester was treated with LiOH-H2O (3-5 equivalents) in MeOH (1-3 mL / mmol ester) and water (1-3 mL / mmol ester) at room temperature for 1-5 hours. The reaction mixture was acidified to pH 4-5 with 1N HCl and concentrated. The residue was purified by preparative HPLC to obtain the desired carboxylic acid product.

[0086] Analysis method LCMS analysis method The final compound was analyzed using LC / MS conditions with a UV detector monitoring at 214 nm and 254 nm, and mass spectrometry scanning at 110–800 amu in ESI+ ionization mode. LC / MS A: Column: XBridge C18, 4.6 × 50 mm, 3.5 μm; Mobile phase: A Water (10 mM ammonium bicarbonate), B CH3CN; Gradient: 5% to 95% B over 1.4 min, then maintained for 1.6 min; Flow rate: 1.8 mL / min; Oven temperature: 50°C. LC / MS B: Column: SunFire C18, 4.6×50mm, 3.5μm; Mobile phase: A Water (0.01% TFA), B CH3CN; Gradient: 5% to 95% B over 1.5 min, then maintained for 1.5 min; Flow rate: 2.0 mL / min; Oven temperature: 50°C. LC / MS C: Column: XBridge C18, 4.6×50mm, 3.5μm, Mobile phase: A Water (10mM ammonium bicarbonate), B CH3CN, Gradient: 5%~95%B over 1.5 min, then maintained for 1.5 min, Flow rate: 1.8 mL / min, Oven temperature 50°C. LC / MS D: Column: Poroshell 120 EC-C138, 4.6 × 30 mm, 2.7 μm; Mobile phase: A Water (0.01% TFA), B CH3CN (0.01% TFA); Gradient: 5% to 95% B over 1.2 min, then maintained for 1.8 min; Flow rate: 2.2 mL / min; Oven temperature: 50°C.

[0087] [Example 2A] Preparation of intermediates Preparation of ethyl(3S)-3-amino-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate Step 1: 2,6-dibromo-4-fluoro-3-methylaniline

[0088] [ka] To a mixture of 4-fluoro-3-methylaniline (50.0 g, 400 mmol) in MeOH (120 mL) and DCM (120 mL), bromine (52 mL, 1.0 mol) was added dropwise at room temperature over 1.5 hours, and the mixture was stirred at room temperature for 4 hours. 1N Na2S2O3 aqueous solution (300 mL) and ethyl acetate (500 mL) were added, and the mixture was stirred for 10 minutes. Then, the mixture was carefully basicized with 1N Na2CO3 aqueous solution (300 mL). The organic layer was separated, and the aqueous layer was extracted with ethyl acetate (2 × 100 mL). The combined organic layers were washed with 1N Na2S2O3 aqueous solution (300 mL) and saline solution (200 mL), dehydrated with Na2SO4, filtered, and concentrated under vacuum to provide 2,6-dibromo-4-fluoro-3-methylaniline as a white solid (80 g). Yield 70.7% (ESI 284.0 [M+H]) + ).

[0089] Step 2: 4-Fluoro-2,3,6-trimethylaniline

[0090] [ka] To a solution of ethyl 2,6-dibromo-4-fluoro-3-methylaniline (50.0 g, 273 mmol) in dioxane (500.0 mL) and water (50 mL), methylboronic acid (49.0 g, 819 mmol), K2CO3 (111.0 g, 819 mmol), and 1,1'-bis(diphenylphosphino)ferrocene-palladium(II) dichloride dichloromethane complex (10.0 g, 13.65 mmol) were added. The mixture was stirred overnight at 110°C. The reaction product was poured into water (500 mL) and extracted with ethyl acetate (500 mL). The combined organic layer was dehydrated with anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel column (petroleum ether: HCl 1:5) to provide 4-fluoro-2,3,6-trimethylaniline as a colorless oil (20.0 g). Yield 47.6% (ESI 154.3 (M+H)) + )

[0091] Step 3: 2-Bromo-5-fluoro-1,3,4-trimethylbenzene

[0092] [ka] To a mixture of 4-fluoro-2,3,6-trimethylaniline (3.8 g, 24.8 mmol) in MeCN (30 mL), t-BuONO (3.8 g, 37.2 mmol) was added at 0°C, followed by the addition of CuBr (4.3 g, 29.7 mmol). The mixture was stirred at 60°C for 2 hours. The reaction mixture was concentrated under vacuum, and the residue was purified by silica gel column (petroleum ether) to provide 2-bromo-5-fluoro-1,3,4-trimethylbenzene as a colorless oil (1.3 g). Yield 33.9%.

[0093] Step 4: Ethyl(3S)-3-(((R)-tert-butylsulfinyl)amino)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate

[0094] [ka] A mixture of (3-((S)-1-(((R)-tert-butylsulfinyl)amino)-3-ethoxy-3-oxopropyl)-2,4-difluoro-5-(trifluoromethyl)phenyl)boronic acid (1.5 g, 3.3 mmol), 2-bromo-5-fluoro-1,3,4-trimethylbenzene (950 mg, 4.3 mmol), K3PO4 (2.1 g, 9.9 mmol), and X-PhosPdG2 (285 mg, 0.33 mmol) in dioxane (10 mL) and H2O (2 mL) was stirred at 110°C for 2 hours under an N2 atmosphere. The mixture was poured into water (100 mL) and extracted with ELISA (100 mL x 3). The combined organic layer was dehydrated with anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (petroleum ether: ethyl 1:1) to provide ethyl(3S)-3-(((R)-tert-butylsulfinyl)amino)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate (380 mg) as a yellow oil. Yield 21.3% (ESI 538.0(M+H)). + ).

[0095] Step 5: Ethyl(3S)-3-amino-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate

[0096] [ka] To a solution of ethyl(3S)-3-(((R)-tert-butylsulfinyl)amino)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate (380 mg, 0.70 mmol) in DCM (5 mL), HCl-dioxane (4 M, 5 mL) was added and the mixture was stirred at room temperature for 1 hour. The mixture was concentrated under vacuum to provide ethyl(3S)-3-amino-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate as a yellow oil (340 mg), which was used directly in the next step without further purification. Yield 100% (ESI 434.2 [M+H]) + ).

[0097] Preparation of ethyl(S)-3-amino-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethyl-[1,1'-biphenyl]-3-yl)propanoate Step 1: (S)-Ethyl 3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)-3-((R)-1,1-dimethylethylsulfinamide)propanoate

[0098] [ka] X-PhosPdG2 (79 mg, 0.1 mmol) was added to a mixture of (S)-ethyl 3-(3-bromo-5-cyclopropyl-2,6-difluorophenyl)-3-((R)-1,1-dimethylethylsulfinamide)propanoate (600 mg, 1.3 mmol), mesitylboronic acid (246 mg, 1.5 mmol), and K3PO4 (848 mg, 43.0 mmol) in dioxane (10 mL) and H2O (1 mL). The mixture was heated at 110 °C under a nitrogen atmosphere for 2 hours. Water (20 mL) was added, and the solution was extracted with ELISA (20 mL x 3). The combined organic phases were concentrated under vacuum, and the residue was purified by silica gel column chromatography (petroleum ether: ELISA 2:1) to provide (S)-ethyl 3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)-3-((R)-1,1-dimethylethylsulfinamide)propanoate as a dark solid (400 mg). Yield 80% (ESI 492.1 [M-100+H]). + )

[0099] Step 2: Ethyl(S)-3-amino-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethyl-[1,1'-biphenyl]-3-yl)propanoate

[0100] [ka] To a mixture of (S)-ethyl 3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)-3-((R)-1,1-dimethylethylsulfinamide)propanoate (550 mg, 1.12 mmol) in DCM (1 mL) and EtOH (2 mL), 4 M HCl-dioxane (2 mL, 5.0 mmol) was added, and the mixture was stirred at room temperature for 0.5 hours. The mixture was then concentrated under vacuum to provide ethyl (S)-3-amino-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethyl-[1,1'-biphenyl]-3-yl)propanoate as a yellow oil (360 mg, crude), which was used directly in the following reaction. (ESI 388.1(M+H))+ )。

[0101] Preparation of Ethyl (3S)-3-amino-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate Step 1: Ethyl (S)-3-(((R)-tert-butylsulfinyl)amino)-3-(3-cyclopropyl-2,6-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoate

[0102]

Chem.

[0103] Step 2: Ethyl(3S)-3-(((R)-tert-butylsulfinyl)amino)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate

[0104] [ka] A mixture of ethyl(S)-3-(((R)-tert-butylsulfinyl)amino)-3-(3-cyclopropyl-2,6-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)phenyl)propanoate (500 mg, 1.0 mmol), K3PO4 (636 mg, 3.0 mmol), Xphos-PdG2 (78.7 mg, 0.1 mmol), and 2-bromo-4-fluoro-1,3-dimethylbenzene (263 mg, 1.3 mmol) in dioxane (10 mL) and H2O (1 mL) was stirred at 110 °C under a nitrogen atmosphere for 2 hours. The reaction mixture was poured into 15 mL of water and extracted with ethyl(15 × 3 mL × 3). The combined organic layer was washed with saline solution (5 mL), dehydrated with anhydrous Na2SO4, filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (petroleum ether: ELISA 3:1) to provide ethyl(3S)-3-(((R)-tert-butylsulfinyl)amino)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate as a colorless oil (200 mg). Yield 40% (ESI 496.2 (M+H)). + )

[0105] Step 3: Ethyl(3S)-3-amino-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate

[0106] [ka] A mixture of ethyl(3S)-3-(((R)-tert-butylsulfinyl)amino)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate (200 mg, 0.4 mmol) in EtOH (2 mL) was mixed with HCl-dioxane (4 M, 2 mL, 8.0 mmol). The mixture was stirred at room temperature for 1 hour. The mixture was concentrated under vacuum, and the residue was filtered through a C18 / 40 g column (A: water / 0.01% TFA). 、 Ethyl (3S)-3-amino-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate was purified by reverse-phase HPLC over B:MeOH (0-100%) to provide 120 mg of oil. Yield 76% (ESI 392.2 [M+H]). + ).

[0107] [Example 2B] Preparation of intermediates Preparation of 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid

[0108] Step 1: (E)-2-methoxy-5-(2-methoxyvinyl)-4-(trifluoromethyl)pyridine

[0109] [ka] Potassium tert-butoxide (376 mg, 3.35 mmol) was added to a solution of (methoxymethyl)triphenylphosphonium chloride (1.0 g, 2.95 mmol) in THF (13.406 mL) at 0°C. After stirring at 0°C for 1 hour, a solution of 6-methoxy-4-(trifluoromethyl)nicotinaldehyde (550 mg, 2.68 mmol) in THF (6.5 mL) was added. The reaction mixture was stirred overnight at room temperature and the reaction was stopped with NH4Cl solution. The mixture was extracted with (Â×3), concentrated, and purified by silica gel chromatography (0-100 ethyl acetate:hexane) to obtain (E)-2-methoxy-5-(2-methoxyvinyl)-4-(trifluoromethyl)pyridine (450 mgs). Yield 72% (ESI 234.2(M+H) + ).

[0110] Step 2: 2-(6-methoxy-4-(trifluoromethyl)pyridine-3-yl)acetaldehyde

[0111] [ka] To a solution of (E)-2-methoxy-5-(2-methoxyvinyl)-4-(trifluoromethyl)pyridine (450 mg, 1.930 mmol) in DCM (29.689 mL), TFA (0.595 mL, 7.72 mmol) and water (0.591 mL, 32.8 mmol) were added. The reaction mixture was stirred at 45°C for 18 hours. The reaction mixture was diluted with DCM and stopped with NaHCO3. The mixture was washed with water, dried over Na2SO4, filtered, and concentrated to obtain 2-(6-methoxy-4-(trifluoromethyl)pyridine-3-yl)acetaldehyde, which was used without further purification (343 mg). Yield 81% (ESI 220.18(M+H)). + ).

[0112] Step 3: 5-(2-(azetidine-1-yl)ethyl)-2-methoxy-4-(trifluoromethyl)pyridine

[0113] [ka] Azetidine hydrochloride (3.4 g, 36.2 mmol) was added to a solution of 2-(6-methoxy-4-(trifluoromethyl)pyridine-3-yl)acetaldehyde (4 g, 18.1 mmol) in DCE (50 mL). The reaction mixture was stirred at room temperature for 20 minutes. NaBH(OAc)3 (7.7 g, 36.2 mmol) was added, and the mixture was stirred at room temperature for 16 hours. The reaction mixture was stopped by adding MeOH (20 mL), and the mixture was filtered. The filtrate was concentrated under vacuum, and the residue was purified by reverse-phase HPLC (A: water, 10 mM NH4HCO3, B: MeOH, 0-100%) on a C18 / 80 g column to obtain 5-(2-(azetidine-1-yl)ethyl)-2-methoxy-4-(trifluoromethyl)pyridine as a yellow oil (3 g). Yield 63% (ESI 261.2(M+H)). + ).

[0114] Step 4: 5-(2-(azetidine-1-yl)ethyl)-4-(trifluoromethyl)pyridine-2(1H)-one

[0115] [ka] A mixture of 5-(2-(azetidine-1-yl)ethyl)-2-methoxy-4-(trifluoromethyl)pyridine (2.95 g, 11.3 mmol) in HBr / AcOH (20 mL) was stirred at 50°C for 5 hours. The reaction mixture was concentrated under vacuum, and the residue was purified by reverse-phase HPLC (A: water, 10 mM NH4HCO3, B: MeOH, 0-100%) on a C18 / 80 g column to obtain 5-(2-(azetidine-1-yl)ethyl)-4-(trifluoromethyl)pyridine-2(1H)-one as a yellow oil (710 mg). Yield 25% (ESI 247.1(M+H)). + ).

[0116] Step 5: Ethyl 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate

[0117] [ka] To a solution of 5-(2-(azetidine-1-yl)ethyl)-4-(trifluoromethyl)pyridine-2(1H)-one (710 mg, 2.9 mmol) in MeCN (10 mL), ethyl 4-methyl-2-(methylsulfonyloxy)pentanoate (1.1 g, 4.4 mmol) and K2CO3 (1.2 g, 8.7 mmol) were added. The reaction mixture was stirred at 80°C for 16 hours. The reaction mixture was concentrated under vacuum, and the residue was purified by reverse-phase HPLC (A: water, 10 mM NH4HCO3, B: MeOH, 0-100%) on a C18 / 40 g column to obtain ethyl 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate as a yellow oil (500 mg). Yield 44% (ESI 389.2(M+H)) + ).

[0118] Step 6: 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid

[0119] [ka] Ethyl 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate (500 mg, 1.3 mmol) was treated with LiOH-H2O (270 mg, 6.5 mmol) in EtOH (5 mL) and water (1 mL) at room temperature for 2 hours. The reaction mixture was neutralized with 2N HCl and concentrated under vacuum. The residue was purified by reverse-phase HPLC on a C18 / 40 g column (A: water, 10 mM NH4HCO3, B: MeOH, 0-100%) to obtain 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid as a yellow oil (410 mg). Yield 88% (ESI 361.2(M+H))+ ).

[0120] Preparation of 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid

[0121] Step 1: 2-(6-methoxy-4-(trifluoromethyl)pyridine-3-yl)-N,N-dimethylethane-1-amine

[0122] [ka] To a solution of 2-(6-methoxy-4-(trifluoromethyl)pyridine-3-yl)acetaldehyde (0.34 g, 1.6 mmol) in DCE (7.8 mL), dimethylamine (3.9 mL, 7.8 mmol) and acetic acid (0.05 mL, 0.78 mmol) were added, and the mixture was stirred for 1 hour. Sodium triacetoxyborohydride (0.6 g, 3.1 mmol) was added to the solution. The reaction mixture was stirred for 12 hours, then concentrated, and purified by silica gel chromatography (0-35% DCM (1% TEA): MeOH 0-30%) to obtain 2-(6-methoxy-4-(trifluoromethyl)pyridine-3-yl)-N,N-dimethylethane-1-amine (305 mg). Yield 79% (ESI 249.27 (M+H)). + ).

[0123] Step 2: 5-(2-(dimethylamino)ethyl)-4-(trifluoromethyl)pyridine-2(1H)-one

[0124] [ka] HBr (33% in acetic acid) (4.04 mL, 24.57 mmol) was added to 2-(6-methoxy-4-(trifluoromethyl)pyridine-3-yl)-N,N-dimethylethane-1-amine (0.305 g, 1.229 mmol) and heated to 75°C in a pressure vessel. After 4 hours, the solvent was removed, and the residue was purified by silica gel chromatography (0-25% DCM:MeOH, with 1% TEA as a modifier) ​​to obtain 5-(2-(dimethylamino)ethyl)-4-(trifluoromethyl)pyridine-2(1H)-one (219 mg). Yield 76% (ESI 235.15(M+H)). + ). 1 H NMR (400 MHz, MeOD) δ 7.56 (s, 1H), 6.85 (s, 1H), 2.76 (m, 2H), 2.61 (m, 1H), 2.37 (m, 6H)

[0125] Step 3: Ethyl 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate

[0126] [ka] A mixture of 5-(2-(dimethylamino)ethyl)-4-(trifluoromethyl)pyridine-2(1H)-one (685 mg, 2.92 mmol), K2CO3 (1.60 g, 11.55 mmol), and ethyl 4-methyl-2-(methylsulfonyloxy)pentanoate (1.60 g, 6.70 mmol) in CH3CN (60 mL) was stirred overnight at 85°C. The solvent was concentrated under vacuum, and the residue was purified by silica gel column (DCM:MeOH 2:1) to obtain ethyl 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate as a brown oil (390 mg). Yield 35% (ESI 377.2 (M+H)). + ). 1H NMR (500 MHz, MeOD) δ 7.84 (s, 1H), 6.68 (s, 1H), 5.51 (dd, J = 11.0, 5.0 Hz, 1H), 4.23 (q, J = 7.0 Hz, 2H), 2.77 (t, J = 8.0 Hz, 2H), 2.53 (t, J = 8.0 Hz, 2H), 2.33 (s, 6H), 2.18 - 2.12 (m, 1H), 2.08 - 2.02 (m, 1H), 1.46 - 1.38 (m, 1H), 1.27 (t, J = 7.0 Hz, 3H), 0.97 (t, J = 7.0 Hz, 6H).

[0127] Step 4: 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid

[0128] [ka] Ethyl 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate (390 mg, 1.0 mmol) was treated with LiOH monohydrate (435 mg, 10.36 mmol) in EtOH (10 mL) and H2O (1 mL) at room temperature for 1 hour. The mixture was acidified to pH 4-5 with 1N HCl aqueous solution. The mixture was concentrated under vacuum and purified by silica gel column (MeOH:siRNA 1:2) to obtain 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid as oil (358 mg). Yield 99% (ESI 349.1(M+H)) + ).

[0129] Preparation of 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid Step 1: Ethyl 2-(5-(3-(dimethylamino)propa-1-in-1-yl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate

[0130] [ka] To a solution of ethyl 2-(5-bromo-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate (68.0 g, 177 mmol, 1.00 equivalent) in THF (408 mL), N,N-dimethylpropa-2-in-1-amine (19.1 g, 230 mmol, 24.4 mL, 1.3 equivalent), CuI (3.37 g, 17.7 mmol, 0.10 equivalent), Pd(PPh3)2Cl2 (6.21 g, 8.85 mmol, 0.05 equivalent), and TEA (358 g, 3.54 mol, 493 mL, 20 equivalent) were added. The reaction mixture was stirred at 25°C for 12 hours. The reaction mixture was diluted with ethyl acetate and washed twice with saturated ammonium chloride solution and once with saline solution. The combined organic phases were dehydrated with sodium sulfate, filtered, and concentrated under reduced pressure. The crude product was purified by silica gel chromatography (petroleum ether:ethyl acetate = 3:1) to obtain ethyl 2-(5-(3-(dimethylamino)propa-1-in-1-yl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate (20.0 g, 51.8 mmol, 29.2% yield) as a yellow oil.

[0131] Step 2: Ethyl 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate

[0132] [ka] To a solution of ethyl 2-(5-(3-(dimethylamino)propane-1-in-1-yl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate (20.0 g, 51.8 mmol, 1 equivalent) in EtOH (200 mL), Pd / C (6.00 g, 2.59 mmol, 5% purity, 0.05 equivalent) was added. The mixture was stirred at 15°C for 24 hours under H2 (50 psi). The reaction mixture was filtered, and the filtrate was concentrated to obtain ethyl 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate (19.7 g, 44.4 mmol, 85.7% yield) as a yellow oil.

[0133] Step 3: 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid

[0134] [ka] To a solution of ethyl 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate (19.7 g, 50.4 mmol, 1 equivalent) in THF (98.0 mL) and H2O (20.0 mL), LiOH.H2O (4.23 g, 101 mmol, 2.00 equivalent) was added at 0°C. The reaction mixture was heated to 20°C and stirred at 20°C for 12 hours. The reaction mixture was adjusted to pH=7 with 1N HCl aqueous solution and concentrated under vacuum to obtain the crude product. The crude product was subjected to prep-HPLC (column: Phenomenex luna C18 (250)). * The mixture was purified by (70 mm, 10 μm); mobile phase: [water (0.05% HCl)-ACN]; B%: 15%~45%, 20 min) to provide 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid (5.40 g, 13.5 mmol, 26.8% yield, HCl) as a white solid. 1H NMR: 400 MHz D2O δ: 7.71 (s, 1H), 7.00 (s,1H), 5.40-5.45 (d, J = 20 Hz, 1H), 3.13-3.17 (m, 2H), 2.84 (s, 6H), 2.65-2.67 (m, 2H), 1.95-1.99 (m, 4H), 1.25-1.30 (m, 1H), 0.86 (s, 9H)

[0135] [Example 3] Synthesis of Exemplary Compounds of the Present Invention Preparative HPLC method Crude samples were dissolved in MeOH and purified by preparative HPLC using a Gilson 215 instrument. Detection wavelength: 214 nm. Preparative HPLC A: Column: Xtimate C18, 21.2 × 250 mm, 10 μm; Mobile phase: A Water (10 mM ammonium bicarbonate), B CH3CN; Gradient elution as described in the text; Flow rate: 30 mL / min. Preparative HPLC B: Column: Xtimate C18, 21.2 × 250 mm, 10 μm, Mobile phase: A Water (0.1% formic acid), B CH3CN, Gradient elution as described in the text, Flow rate: 30 mL / min.

[0136] 3-52. Preparation of (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoic acid (compounds HH-P1 and HH-P2) Step 1: Ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate

[0137] [ka] A mixture of ethyl(3S)-3-amino-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate (120 mg, 0.3 mmol), 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid (108 mg, 0.3 mmol), TCFH (126 mg, 0.45 mmol), and NMI (123 mg, 1.5 mmol) in CH3CN (3 mL) was stirred overnight at room temperature. The reaction mixture was concentrated under vacuum, and the residue was analyzed on a C18 / 40 g column (A: 10 mM water NH4HCO3). 3、 Ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate was provided as a pale yellow solid (140 mg). Yield 62% (ESI 734.2 [M+H]). + ).

[0138] Step 2: (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoic acid

[0139] [ka] Ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate (140 mg, 0.19 mmol) was treated with LiOH-H2O (24 mg, 0.57 mmol) in EtOH (2 mL) and water (0.5 mL) at room temperature for 1 hour. The reaction mixture was acidified to pH 4-5 with 2N HCl. The reaction product was concentrated under vacuum, and the residue was purified by prep-HPLC A (20-85% MeCN) to obtain diastereomer products HH-P1 (35 mg) and HH-P2 (58 mg) as white solids. HH-P1 ESI 706.2 (M+H) + 1 H NMR (400 MHz, MeOD) δ 7.80 (s, 1H), 7.12 - 7.03 (m, 1H), 7.00 - 6.91 (m, 1H), 6.87 (s, 1H), 6.62 (t, J = 8.0 Hz, 1H), 5.80 - 5.58 (m, 2H), 4.00 (t, J = 7.7 Hz, 4H), 3.30 - 3.19 (m, 2H), 2.97 - 2.78 (m, 3H), 2.71 - 2.62 (m, 1H), 2.49 - 2.34 (m, 2H), 2.13 - 1.76 (m, 9H), 1.46 - 1.33 (m, 1H), 1.00 - 0.86 (m, 8H), 0.66 (d, J = 4.8 Hz, 2H). HH-P2 ESI 706.2 (M+H) + 1H NMR (400 MHz, MeOD) δ 7.70 (s, 1H), 7.14 - 7.04 (m, 1H), 7.02 - 6.87 (m, 2H), 6.67 (t, J = 8.1 Hz, 1H), 5.98 - 5.86 (m, 1H), 5.62 (t, J = 7.7 Hz, 1H), 4.11 (t, J = 8.1 Hz, 4H), 3.45 - 3.33 (m, 2H), 2.99 - 2.71 (m, 3H), 2.60 - 2.36 (m, 3H), 2.17 - 2.03 (m, 1H), 2.00 - 1.86 (m, 7H), 1.80 - 1.67 (m, 1H), 1.41 - 1.29 (m, 1H), 1.06 - 0.94 (m, 2H), 0.93 - 0.82 (m, 6H), 0.72 - 0.62 (m, 2H).

[0140] 3-57. Preparation of (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propanoic acid (compounds HM-P1 and HM-P2) Step 1: (3S)-Ethyl 3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propanoate

[0141] [ka] A mixture of (S)-ethyl 3-amino-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propanoate (120 mg, 0.31 mmol), 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid (112 mg, 0.31 mmol), TCFH (174 mg, 0.62 mmol), and NMI (82 mg, 1.0 mmol) in MeCN (5 mL) was stirred at room temperature for 1 hour. The solvent was removed under vacuum, and the residue was purified by silica gel column (DCM:MeOH 97:3) to provide (3S)-ethyl 3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propanoate as a colorless oil (120 mg). Yield 53% (ESI 730.3 (M+H)). + ).

[0142] Step 2: (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propanoic acid

[0143] [ka] (3S)-ethyl 3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propanoate (120 mg, 0.16 mmol) was treated with EtOH (3 mL) and LiOH-H2O (35 mg, 0.9 mmol) in H2O (1 mL) at room temperature for 2 hours. The reaction mixture was acidified to pH 4-5 with 1N HCl. The reaction mixture was concentrated under vacuum, and the residue was purified by prep-HPLC A (30-60% CH3CN) to obtain diastereomer products HM-P1 (30.2 mg) and HM-P2 (31.8 mg) as white solids. HM-P1 ESI 702.3 (M+H) + .1H NMR (400 MHz, MeOD) δ 7.80 (s, 1H), 6.97 - 6.79 (m, 3H), 6.58 (t, J = 8.1 Hz, 1H), 5.84 - 5.61 (m, 2H), 4.00 (t, J = 8.1 Hz, 4H), 3.30 - 3.21 (m, 2H), 2.88-2.85 (m, 3H), 2.67 (dd, J = 14.8, 4.8 Hz, 1H), 2.48 - 2.37 (m, 2H), 2.28 (d, J = 6.4 Hz, 3H), 2.13 - 1.96 (m, 3H), 1.90 (d, J = 21.3 Hz, 6H), 1.46 - 1.31 (m, 1H), 0.94 -0.90(m, 8H), 0.64-0.62 (m, 2H). HM-P2 ESI 702.3 (M+H) +.1H NMR (400 MHz, MeOD) δ 7.70 (s, 1H), 7.00 - 6.86 (m, 3H), 6.63 (t, J = 8.2 Hz, 1H), 5.93 (dd, J = 11.4, 3.5 Hz, 1H), 5.62 (t, J = 7.7 Hz, 1H), 4.11 (t, J = 8.0 Hz, 4H), 3.50 - 3.33 -3.30(m, 2H), 2.97 - 2.76 (m, 3H), 2.47-2.45 (m, 3H), 2.29 (s, 3H), 2.08-2.05 (m, 1H), 2.00 - 1.89 (m, 7H), 1.74-1.70 (m, 1H), 1.37 -1.35(m, 1H), 1.04 - 0.95 (m, 2H), 0.88 (dd, J = 11.4, 6.6 Hz, 6H), 0.67-0.65 (m, 2H).

[0144] 3-62. Preparation of (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoic acid (compounds HR-P1 and HR-P2) Step 1: Ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate

[0145] [ka] A mixture of ethyl(3S)-3-amino-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate (90 mg, 0.21 mmol), 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid (81 mg, 0.21 mmol), NMI (0.2 mL), and TCFH (88 mg, 0.32 mmol) in CH3CN (3 mL) was stirred at room temperature for 2 hours. The solvent was concentrated under vacuum, and the residue was purified by reverse-phase HPLC on a C18 / 40g column (A: 10mM water NH4HCO3, B: CH3CN, 0-100%) to provide ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate as a white solid (80 mg). Yield 50% (ESI 776.2 [M+H]). + ).

[0146] Step 2: (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoic acid

[0147] [ka] Ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate (80 mg, 0.1 mmol) was treated with LiOH-H2O (13 mg, 0.3 mmol) in MeOH (2.0 mL) and H2O (0.5 mL) at room temperature for 2 hours. The reaction mixture was acidified to pH 5-6 with 1N HCl. The reaction products were concentrated under vacuum, and the residue was purified by prep-HPLC A (50-80% CH3CN) to obtain diastereomer products HR-P1 (22.0 mg) and HR-P2 (25.0 mg) as white solids. HR-P1 ESI 748.2 (M+H) + . 1 H NMR (400 MHz, MeOD) δ 7.79 (s, 1H), 7.40 (t, J = 7.5 Hz, 1H), 6.99 (d, J = 7.9 Hz, 1H), 6.85 (d, J = 3.0 Hz, 1H), 5.78 - 5.60 (m, 2H), 4.04 (t, J = 8.0 Hz, 4H), 3.29 - 3.20 (m, 2H), 2.95 - 2.80 (m, 3H), 2.77 - 2.68 (m, 1H), 2.51 - 2.37 (m, 2H), 2.25 (d, J = 1.6 Hz, 3H), 2.09 - 1.75 (m, 8H), 1.37 (s, 1H), 0.93 (t, J = 6.4 Hz, 6H). HR-P2 ESI 748.2 (M+H) + . 1H NMR (400 MHz, MeOD) δ 7.72 (s, 1H), 7.46 (t, J = 7.5 Hz, 1H), 7.03 (d, J = 7.6 Hz, 1H), 6.91 (s, 1H), 5.87 - 5.82 (m, 1H), 5.63 (t, J = 7.7 Hz, 1H), 4.12 (t, J = 7.8 Hz, 4H), 3.45 - 3.34 (m, 2H), 2.95 - 2.74 (m, 3H), ), 2.64 - 2.56 (m, 1H), 2.50 - 2.36 (m, 2H), 2.27 (d, J = 1.5 Hz, 3H), 2.02 - 1.83 (m, 7H), ), 1.78 - 1.65 (m, 1H), ), 1.40 - 1.26 (m, 1H), ), 0.93 - 0.84 (m, 6H).

[0148] 3-65. Preparation of (3S)-3-(4'-cyclopropyl-2,4-difluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoic acid (compounds HU-P1 and HU-P2) Step 1: Ethyl(3S)-3-(4'-cyclopropyl-2,4-difluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-(2(5(3(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoate

[0149] [ka] A mixture of ethyl(S)-3-amino-3-(4'-cyclopropyl-2,4-difluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate (140.0 mg, 0.32 mmol), 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid (116.0 mg, 0.32 mmol), TCFH (106.6 mg, 0.38 mmol), and NMI (131.4 mg, 1.60 mmol) in CH3CN (5 mL) was stirred overnight at room temperature. The reaction mixture was concentrated under vacuum, and the residue was analyzed on a C18 / 40 g column (A: 10 mM water NH4HCO3). 3、 Ethyl (3S)-3-(4'-cyclopropyl-2,4-difluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoate was purified by reverse-phase HPLC over B:MeOH (0-100%) to provide 150.0 mg as a pale yellow solid. Yield 60% (ESI 786.3 [M+H]). + ).

[0150] Step 2: (3S)-3-(4'-Cyclopropyl-2,4-Difluoro-2',6'-Dimethyl-5-(Trifluoromethyl)-[1,1'-Biphenyl]-3-yl)-3-(2(5(3(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-Methylpentanamide)Propanoic Acid

[0151] [ka] Ethyl(3S)-3-(4'-cyclopropyl-2,4-difluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoate (150.0 mg, 0.19 mmol) was treated with LiOH-H2O (23.9 mg, 0.57 mmol) in EtOH (2 mL) and water (0.5 mL) at room temperature for 1 hour. The reaction mixture was acidified to pH 4-5 with 2N HCl. The reaction products were concentrated under vacuum, and the residue was purified by prep-HPLC A (30-80% MeCN) to obtain the diastereomer products HU-P1 (54.5 mg) and HU-P2 (50.5 mg) as white solids. HU-P1 ESI 758.3 (M+H) + 1 H NMR (400 MHz, MeOD) δ 7.76 (s, 1H), 7.34 (t, J = 7.6 Hz, 1H), 6.87 - 6.76 (m, 3H), 5.81 - 5.67 (m, 2H), 3.12 - 3.04 (m, 2H), 2.98 - 2.90 (m, 1H), 2.79 (s, 6H), 2.76 - 2.57 (m, 3H), 2.11 - 1.79 (m, 11H), 1.41 - 1.27 (m, 1H), 0.99 - 0.89 (m, 8H), 0.71 - 0.65 (m, 2H). HU-P2 ESI 758.3 (M+H) + 1H NMR (400 MHz, MeOD) δ 7.79 (s, 1H), 7.39 (t, J = 7.6 Hz, 1H), 6.85 (s, 3H), 5.85 - 5.79 (m, 1H), 5.62 (t, J = 7.6 Hz, 1H), 3.13 - 2.94 (m, 2H), 2.93 - 2.84 (m, 1H), 2.79 (s, 6H), 2.70 - 2.57 (m, 3H), 2.09 - 1.81 (m, 10H), 1.70 - 1.58 (m, 1H), 1.36 - 1.26 (m, 1H), 0.99 - 0.92 (m, 2H), 0.89 - 0.83 (m, 6H), 0.72 - 0.65 (m, 2H).

[0152] 3-67. Preparation of (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoic acid (compounds HW-P1 and HW-P2) Step 1: Ethyl(3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate

[0153] [ka] A mixture of 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid (100.0 mg, 0.28 mmol), ethyl(S)-3-amino-3-(2,4,4'-trifluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate (117.3 mg, 0.28 mmol), TCFH (156.8 mg, 0.56 mmol), and NMI (91.8 mg, 1.12 mmol) in CH3CN (10 mL) was stirred at room temperature for 1 hour. The reaction mixture was concentrated under vacuum, and the residue was purified by silica gel column (DCM:MeOH 4:1) to provide ethyl(3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate as a brown solid (150.0 mg). Yield 71% (ESI 764.7 (M+H)). + ).

[0154] Step 2: (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoic acid

[0155] [ka] Ethyl(3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate (150 mg, 0.20 mmol)) was treated with LiOH-H2O (33.6 mg, 0.80 mmol) in MeOH (4 mL) and H2O (1 mL) at room temperature for 2 hours. The reaction mixture was acidified to pH 4-5 with 1N HCl. The reaction product was concentrated under vacuum, and the residue was purified by prep-HPLC A (30-60% MeCN) to obtain diastereomer products HW-P1 (60.1 mg) and HW-P2 (50.0 mg) as white solids. HW-P1 ESI 736.6 (M+H) + .1H NMR (400 MHz, MeOD) δ 7.76 (s, 1H), 7.39 (t, J = 7.5 Hz, 1H), 6.88 (d, J = 9.3 Hz, 2H), 6.80 (s, 1H), 5.78 - 5.67 (m, 2H), 3.14 - 3.02 (m, 2H), 3.01 - 2.88 (m, 1H), 2.80 (s, 6H), 2.75 - 2.55 (m, 3H), 2.12 - 1.84 (m, 10H), 1.34 (s, 1H), 0.94 (d, J = 6.6 Hz, 6H). HW-P2 ESI 736.6 (M+H) +.1H NMR (400 MHz, MeOD) δ 7.79 (s, 1H), 7.45 (t, J = 7.6 Hz, 1H), 6.96 - 6.80 (m, 3H), 5.86 - 5.75 (m, 1H), 5.62 (t, J = 7.6 Hz, 1H), 3.14 - 2.97 (m, 2H), 2.97 - 2.85 (m, 1H), 2.79 (s, 6H), 2.71 - 2.50 (m, 3H), 2.07 - 1.87 (m, 8H), 1.67 - 1.52 (m, 1H), 1.35 - 1.25 (m, 1H), 0.93 - 0.77 (m, 6H).

[0156] 3-70. Preparation of (3S)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoic acid (compounds HZ-P1 and HZ-P2) Step 1: (3S)-Ethyl 3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoate

[0157] [ka] A mixture of (S)-ethyl 3-amino-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propanoate (208 mg, 0.50 mmol), 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid (180 mg, 0.50 mmol), TCFH (280 mg, 1.0 mmol), and NMI (123 mg, 1.5 mmol) in MeCN (5 mL) was stirred at room temperature for 3 hours. The solvent was removed under vacuum, and the residue was purified by silica gel column (DCM:MeOH 97:3) to provide (3S)-ethyl 3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoate as a colorless oil (200 mg). Yield 53% (ESI 760.3 (M+H)). + ).

[0158] Step 2: (3S)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoic acid

[0159] [ka] (3S)-ethyl 3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoate (200 mg, 0.26 mmol) was treated with LiOH-H2O (55 mg, 1.3 mmol) in EtOH (3 mL) and H2O (1 mL) at room temperature for 2 hours. The reaction mixture was acidified to pH 4-5 with 1N HCl. The reaction product was concentrated under vacuum, and the residue was purified by prep-HPLC A (30-60% CH3CN) to obtain diastereomer products HZ-P1 (7.0 mg) and HZ-P2 (62.0 mg) as white solids. HZ-P1 ESI 732.3 (M+H) + .1H NMR (400 MHz, MeOD) δ 7.76 (s, 1H), 7.35 (t, J = 7.6 Hz, 1H), 6.94 (s, 2H), 6.80 (s, 1H), 5.79 - 5.70 (m, 2H), 3.14 - 3.07 (m, 2H), 2.95-2.90 (m, 1H), 2.81 (s, 6H), 2.72 - 2.58 (m, 3H), 2.30 (s, 3H), 2.11 - 1.92 (m, 7H), 1.87 (s, 3H), 1.34 (s, 1H), 0.94 (d, J = 6.5 Hz, 6H). HZ-P2 ESI 732.3 (M+H) +.1H NMR (400 MHz, MeOD) δ 7.79 (s, 1H), 7.40 (t, J = 7.6 Hz, 1H), 6.97 (s, 2H), 6.85 (s, 1H), 5.82-5.80 (m, 1H), 5.62 (t, J = 7.6 Hz, 1H), 3.15 - 2.95 (m, 2H), 2.89-2.85 (m, 1H), 2.79 (s, 6H), 2.63 -2.60(m, 3H), 2.31 (s, 3H), 2.08 - 1.83 (m, 9H), 1.72 - 1.58 (m, 1H), 1.31-1.25 (m, 1H), 0.86-0.82 (m, 6H).

[0160] 3-76. Preparation of (3S)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-(2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoic acid (compounds IF-P1 and IF-P2) Step 1: Ethyl(3S)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-(2(5(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoate

[0161] [ka] A mixture of ethyl(S)-3-amino-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate (210 mg, 0.5 mmol), 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid (200 mg, 0.57 mmol), TCFH (364 mg, 1.3 mmol), and NMI (246 mg, 3.0 mmol) in CH3CN (4 mL) was stirred at 50°C for 1 hour. The reaction product was concentrated and purified by reverse-phase HPLC on a C18 / 40g column (A: 10mM water NH4HCO3, B: MeOH, 0-90%) to provide ethyl(3S)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-(2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoate as a yellow solid (330 mg). Yield 88.5% (ESI 746.3 [M+H]). + ).

[0162] Step 2: (3S)-3-(2,4-difluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)-3-(2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoic acid

[0163] [ka] Ethyl(3S)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-(2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)propanoate (330 mg, 0.44 mmol) was treated with LiOH-H2O (56 mg, 1.34 mmol) in MeOH (4 mL) and H2O (1 mL) at room temperature for 30 minutes. The reaction mixture was acidified to pH 5-6 with 1N HCl. The reaction mixture was concentrated under vacuum, and the residue was purified by prep-HPLC A (30-58% CH3CN) to obtain diastereomer products IF-P1 (96 mg) and IF-P2 (94 mg) as white solids. IF-P1 ESI 718.3 (M+H) + . 1 H NMR (400 MHz, MeOD) δ 7.86 (s, 1H), 7.34 (t, J = 7.6 Hz, 1H), 6.94 (d, J = 3.7 Hz, 2H), 6.82 (s, 1H), 5.76 - 5.65 (m, 2H), 3.12 - 3.04 (m, 2H), 2.99 - 2.88 (m, 3H), 2.80 - 2.70 (m, 7H), 2.29 (s, 3H), 2.06 - 1.92 (m, 5H), 1.86 (s, 3H), 1.44 - 1.33 (m, 1H), 0.93 (t, J = 7.0 Hz, 6H). IF-P2 ESI 718.3 (M+H) +.1H NMR (400 MHz, MeOD) δ 7.84 (s, 1H), 7.41 (t, J = 7.6 Hz, 1H), 6.97 (s, 2H), 6.89 (s, 1H), 5.85 - 5.76 (m, 1H), 5.66 (t, J = 7.8 Hz, 1H), 3.25 - 3.09 (m, 2H), 2.96 (t, J = 7.2 Hz, 2H), 2.90 - 2.81 (m, 1H), 2.79 (s, 6H), 2.71 - 2.62 (m, 1H), 2.31 (s, 3H), 1.98 (d, J = 3.1 Hz, 6H), 1.93 - 1.81 (m, 1H), 1.76 - 1.67 (m, 1H), 1.36 - 1.27 (m, 1H), 0.92 - 0.82 (m, 6H).

[0164] 3-77. Preparation of (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propanoic acid (compounds IG-P1 and IG-P2) Step 1: (3S)-ethyl 3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propanoate

[0165] [ka] A mixture of 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid (150 mg, 0.41 mmol), (S)-ethyl 3-amino-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propanoate (173 mg, 0.41 mmol), NMI (0.5 mL), and TCFH (364 mg, 1.30 mmol) in CH3CN (5 mL) was stirred at room temperature for 1 hour. The solvent was concentrated under vacuum, and the residue was purified by prep-HPLC A (30-90% CH3CN) to provide (3S)-ethyl 3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propanoate as a white solid (150 mg). Yield 54% (ESI 758.2 [M+H]). + ).

[0166] Step 2: (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propanoic acid [ka] (3S)-ethyl 3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propanoate (150 mg, 0.19 mmol) was treated with LiOH-H2O (42 mg, 1.00 mmol) in MeOH (4 mL) and H2O (0.4 mL) at room temperature for 2 hours. The reaction mixture was acidified to pH 4-5 with 1N HCl. The reaction mixture was concentrated under vacuum, and the residue was purified by prep-HPLC A (30-70% CH3CN) to obtain diastereomer products IG-P1 (33.0 mg) and IG-P2 (51.0 mg) as white solids. IG-P1 ESI 730.2 (M+H) + . 1 H NMR (400 MHz, MeOD) δ 7.79 (s, 1H), 7.37 (t, J = 7.5 Hz, 1H), 6.95 (s, 2H), 6.85 (s, 1H), 5.79 - 5.65 (m, 2H), 4.03 (t, J = 8.1 Hz, 4H), 3.27 - 3.20 (m, 2H), 3.00 - 2.69 (m, 4H), 2.50 - 2.38 (m, 2H), 2.30 (s, 3H), 2.05 - 1.83 (m, 8H), 1.52 - 1.28 (m, 1H), 0.93 (t, J = 6.4Hz, 6H). IG-P2 ESI 730.2 (M+H) + . 11H NMR (400 MHz, MeOD) δ 7.73 (s, 1H), 7.43 (t, J = 7.6 Hz, 1H), 6.97 (s, 2H), 6.91 (s, 1H), 6.00 - 5.83 (m, 1H), 5.63 (t, J = 7.7 Hz, 1H), 4.11 (t, J = 8.0 Hz, 4H), 3.50 - 3.33 (m, 2H), 2.99 - 2.75 (m, 3H), 2.69 - 2.57 (m, 1H), 2.49 - 2.37 (m, 2H), 2.31 (s, 3H), 2.02 - 1.84 (m, 7H), 1.77 - 1.64 (m, 1H), 1.46 - 1.19 (m, 1H), 1.03 - 0.80 (m, 6H).

[0167] 3-86. Preparation of (3S)-3-(2-(5-(3-(azetidin-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoic acid (Compound IP-P1 and IP-P2) Step 1: Ethyl (3S)-3-(2-(5-(3-(azetidin-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate [Chemical Structure Diagram] A mixture of ethyl(S)-3-amino-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate (112.0 mg, 0.27 mmol), 2-(5-(3-(azetidine-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid (100.0 mg, 0.27 mmol), TCFH (151.2 mg, 0.54 mmol), and NMI (88.6 mg, 1.08 mmol) in CH3CN (8 mL) was stirred at room temperature for 1 hour. The reaction mixture was concentrated under vacuum, and the residue was purified by silica gel column (DCM:MeOH4:1) to provide ethyl(3S)-3-(2-(5-(3-(azetidine-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate as a brown solid (130.0 mg). Yield 63% (ESI 772.6 (M+H)). + ).

[0168] Step 2: (3S)-3-(2-(5-(3-(azetidine-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoic acid [ka] Ethyl(3S)-3-(2-(5-(3-(azetidine-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate (130 mg, 0.17 mmol)) was treated with LiOH-H2O (28.6 mg, 0.68 mmol) in MeOH (4 mL) and H2O (1 mL) at room temperature for 2 hours. The reaction mixture was acidified to pH 4-5 with 1N HCl. The reaction products were concentrated under vacuum, and the residue was purified by prep-HPLC A (30-60% MeCN) to obtain diastereomer products IP-P1 (38.0 mg) and IP-P2 (42.0 mg) as white solids. IP-P1 ESI 744.7 (M+H) + .1H NMR (400 MHz, MeOD) δ 7.76 (s, 1H), 7.34 (d, J = 7.4 Hz, 1H), 6.94 (s, 1H), 6.79 (s, 2H), 5.74 (d, J = 4.1 Hz, 2H), 4.08 (t, J = 7.7 Hz, 4H), 3.21 - 3.11 (m, 2H), 2.97 - 2.89 (m, 1H), 2.71 - 2.58 (m, 3H), 2.45 (s, 2H), 2.30 (s, 3H), 2.06 - 1.91 (m, 5H), 1.83 (d, J = 43.4 Hz, 5H), 1.34 (s, 1H), 0.93 (d, J = 6.5 Hz, 6H). IP-P2 ESI 744.7 (M+H) +.1H NMR (400 MHz, MeOD) δ 7.78 (s, 1H), 7.40 (t, J = 7.7 Hz, 1H), 6.90 (d, J = 55.2 Hz, 3H), 5.87 - 5.78 (m, 1H), 5.61 (t, J = 7.6 Hz, 1H), 4.15 - 3.99 (m, 4H), 3.19 - 2.97 (m, 2H), 2.94 - 2.82 (m, 1H), 2.69 - 2.54 (m, 3H), 2.50 - 2.37 (m, 2H), 2.31 (s, 3H), 2.03 - 1.71 (m, 9H), 1.73 - 1.53 (m, 1H), 1.39 - 1.20 (m, 1H), 0.89 - 0.72 (m, 6H).

[0169] 3-93. Preparation of (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propanoic acid (compounds IW-P1 and IW-P2) Step 1: Ethyl(3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propanoate

[0170] [ka] A mixture of ethyl(3S)-3-amino-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propanoate (170 mg, 0.45 mmol), 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid (151 mg, 0.54 mmol), TCFH (213.2 mg, 0.76 mmol), and NMI (177 mg, 2.16 mmol) in CH3CN (3 mL) was stirred overnight at room temperature. The reaction mixture was concentrated under vacuum, and the residue was analyzed on a C18 / 40 g column (A: 10 mM water NH4HCO3). 3、 Ethyl(3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propanoate was provided as a pale yellow solid (210.0 mg). Yield 65% (ESI 724.2 [M+H]). + ).

[0171] Step 2: (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propanoic acid

[0172] [ka] Ethyl(3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propanoate (210.0 mg, 0.29 mmol) was treated with LiOH-H2O (36.5 mg, 0.87 mmol) in EtOH (2 mL) and water (0.5 mL) at room temperature for 1 hour. The reaction mixture was acidified to pH 4-5 with 2N HCl. The reaction mixture was concentrated under vacuum, and the residue was purified by prep-HPLC A (20-85% MeCN) to obtain diastereomer products IW-P1 (68 mg) and IW-P2 (52 mg) as white solids. IW-P1 ESI 696.2 (M+H) + 1 H NMR (400 MHz, MeOD) δ 7.79 (s, 1H), 6.98 - 6.80 (m, 3H), 5.83 - 5.69 (m, 2H), 3.09 (t, J = 7.9 Hz, 2H), 3.00 - 2.90 (m, 1H), 2.81 (d, J = 0.8 Hz, 6H), 2.73 - 2.58 (m, 3H), 2.26 (d, J = 1.3 Hz, 6H), 2.15 - 1.78 (m, 10H), 1.43 - 1.30 (m, 1H), 1.00 - 0.90 (m, 6H). IW-P2 ESI 696.2 (M+H) + 1 H NMR (400 MHz, MeOD) δ 7.79 (s, 1H), 7.02 - 6.79 (m, 3H), 5.89 - 5.79 (m, 1H), 5.57 (t, J = 7.6 Hz, 1H), 3.09 - 2.95 (m, 2H), 2.91 - 2.71 (m, 7H), 2.70 - 2.47 (m, 3H), 2.32 - 2.19 (m, 6H), 2.07 - 1.83 (m, 9H), 1.67 - 1.55 (m, 1H), 1.37 - 1.27 (m, 1H), 0.90 - 0.80 (m, 6H).

[0173] 3-142. Preparation of (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoic acid (compounds KT-P1 and KT-P2) Step 1: Ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate

[0174] [ka] A mixture of ethyl(3S)-3-amino-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate (140 mg, 0.32 mmol), 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid (116 mg, 0.32 mmol), TCFH (179 mg, 0.64 mmol), and NMI (131 mg, 1.6 mmol) in CH3CN (5 mL) was stirred at room temperature for 2 hours. The solvent was removed under vacuum, and the residue was purified by reverse-phase HPLC on a C18 / 40g column (A: 10mM water NH4HCO3, B: CH3OH, 0-85%) to provide ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate as a yellow solid (140 mg). Yield 56.4% (ESI 776.3 [M+H]). + ).

[0175] Step 2: (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoic acid

[0176] [ka] Ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate (140 mg, 0.18 mmol) was treated with EtOH (4 mL) and LiOH-H2O (22 mg, 0.54 mmol) in H2O (1 mL) at room temperature for 2 hours. The reaction mixture was acidified to pH 5-6 with 1N HCl. The solvent was removed under vacuum, and the residue was purified by prep-HPLC A (30-65% CH3CN) to provide the diastereomer products KT-P1 (24 mg) and KT-P2 (30 mg) as white solids. KT-P1 ESI 748.2 (M+H) + . 1 H NMR (400 MHz, MeOD) δ 7.81 (s, 1H), 7.39 (m, J = 7.5 Hz, 1H), 6.99 - 6.80 (m, 2H), 5.71 (m, J = 9.2, 5.9 Hz, 2H), 4.12 (m, J = 8.1 Hz, 4H), 3.42 - 3.30 (m, 2H), 2.99 - 2.71 (m, 4H), 2.52 - 2.35 (m, 2H), 2.18 (s, 3H), 2.09 - 1.93 (m, 5H), 1.86 (s, 3H), 1.45 - 1.32 (m, 1H), 0.94 (m, J = 6.9 Hz, 6H). KT-P2 ESI 748.2 (M+H) + . 1 H NMR (400 MHz, MeOD) δ 7.72 (s, 1H), 7.44 (m, J = 7.6 Hz, 1H), 7.00 - 6.77 (m, 2H), 5.90 (d, J = 8.1 Hz, 1H), 5.63 (m, J = 7.7 Hz, 1H), 4.12 (m, J = 8.1 Hz, 4H), 3.39 (m, J = 19.1, 13.7 Hz, 2H), 2.86 (m, J = 14.4, 13.3 Hz, 3H), 2.60 (m, J = 15.6, 4.2 Hz, 1H), 2.45 (m, J = 8.1 Hz, 2H), 2.20 (s, 3H), 1.94 (m, J = 21.4, 9.3, 4.8 Hz, 7H), 1.72 (m, J = 14.4, 7.3 Hz, 1H), 1.33 (m, J = 13.4, 6.7 Hz, 1H), 0.88 (m, J = 10.6, 6.6 Hz, 6H).

[0177] Introduction to the in vitro assay described in Examples 4-6 Three in vitro assays were used to examine the α4β7 mechanism processes utilized by cells: 1) ligand: receptor affinity, 2) binding activity of these interactions on the cell surface, and 3) how these interactions are carried out under the applied force. In Example 4, a fluorescence polarization (FP) assay was used to measure compound activity through binding competition with a fluorescein-labeled peptide. In Example 5, the intensity of the compound's action on α4β7 was measured in a cell-based ligand binding assay (LBA) using RPMI 8866 cells incubated with a compound sample in competition with a soluble MAdCAM-1 ligand. In Example 6, the activity of the compound was evaluated in a cell adhesion assay that examines as a mechanism what happens in vivo when cell transport adheres α4β7 to MAdCAM-1 expressing HEV in the intestine during the spillover process. In the cell adhesion assay of Example 6, MAdCAM1-(Fc) was coated onto plastic, and α4β7-expressing cells (RPMI-8866) were adhered to the coated surface in the presence of the test compound. Next, a washing force in buffer was applied to the cells to thereby test the force of their adhesion. Unbound cells were removed, and the remaining adherent cells were quantified.

[0178] [Example 4] Fluorescence Polarization Assay of Compound for α4β7 Binding Compound activity was measured through binding competition with a fluorescein-labeled peptide CRSDTLCGE{Lys(FITC)} using a fluorescence polarization (FP) assay. In this assay, 6.5 nM of integrin α4β7 was incubated with a test compound in 2 mM manganese chloride, 0.1 mM calcium chloride, 20 mM HEPES buffer at pH 7.3, 150 mM sodium chloride, 0.01% Triton X-100, 2% DMSO, and 3 nM of the fluorescein-labeled peptide. The assay was run in a 384-well plate, and the integrin protein was pre-incubated with the test compound at 22 °C for 15 minutes, after which the fluorescein-labeled peptide was added. After adding the fluorescein-labeled peptide, the assay was incubated at 22 °C for 1 hour, and fluorescence polarization was measured. IC50 The values ​​were determined by nonlinear regression and four-parameter curve fitting.

[0179] The FP assay of Example 4 was used to measure the α4β7 inhibitory effect strength of compounds, including a specific compound shown in Figure 1. In Figure 1, the IC257 obtained using the FP assay of Example 4 was measured. 50 The values ​​are provided as numerical ranges (in Figure 1, A: <5nM; B: 5~500nM; C: >500nM).

[0180] Furthermore, the α4β7 inhibitory intensity was measured for the compounds in Table 1 and (comparative) Table 2 below using the FP assay of Example 4, and the result was the IC obtained. 50 The values ​​are provided as numerical ranges (in Tables 1 and 2, A: ≤ 10nM; B: > 10 to 500nM; C: > 500nM).

[0181] [Example 5] Ligand Binding Assay In cell-based ligand binding assays (LBAs), to measure the efficacy of compounds against α4β7, RPMI8866 cells were incubated for 15 minutes at room temperature with a 10 μl volume of compound sample in a buffer containing 50 mM HEPES, 150 mM sodium chloride, 1% bovine serum albumin, 3 mM manganese chloride, 0.15 mM calcium chloride, 15 mM glucose, 1.5% dimethyl sulfoxide, and 0.025% e780 Fixable Viability Dye at pH 7.3. 5 μl of 33 nM MAdCAM-1-Fc, fluorescently labeled with Dylight 650, was added to the cells in 50 mM HEPES, 150 mM sodium chloride, and 1% bovine serum albumin at pH 7.3. Samples were incubated at room temperature for 45 minutes, fixed with 0.8% formaldehyde at room temperature for 30 minutes, and washed with 50 mM Tris, 150 mM NaCl, 1 mM EDTA, and 1% bovine serum albumin at pH 7.5. Fluorescence intensity for each cell was measured via flow cytometry. Dead cells were excluded from further analysis based on staining with 780 Fixable Viability Dye. The median fluorescence intensity for Dylight 650 was determined for each sample, and the concentration-response curves were analyzed using 4-parameter nonlinear regression analysis (IC). 50 The values ​​were analyzed.

[0182] The compounds listed in Figure 1 were subjected to α4β7 ligand binding assay measurements using the ligand binding assay of Example 5. In Figure 1, the IC2016 50 The values ​​are provided by numerical ranges (in Figure 1, A: <5nM; B: 5~500nM; C: >500nM).

[0183] Furthermore, the compounds in Table 1 and (Comparison) Table 2 below were subjected to α4β7 ligand binding assay measurements using the ligand binding assay of Example 5, and the resulting IC50 was obtained. 50 The values ​​are provided as numerical ranges (in Tables 1 and 2, A: ≤ 10nM; B: > 10 to 500nM; C: > 500nM).

[0184] [Example 6] Cell adhesion assay Example 6 describes a cell adhesion assay. α4β7 cell adhesion measurements from the assay in Example 6 were obtained using the compounds listed in Table 1 and similarly using the comparative compounds in Table 2. The results were obtained using the resulting IC. 50 The values ​​are displayed as numerical ranges (for Tables 1 and 2, A: <5nM; B: 5 to <10nM; C: 10 to 50nM; D: >50nM; E: >100nM and F: >500nM).

[0185] Add 100 μg of recombinant human MAdCAM in 100 μl of PBS to each well of a 96-well plate and incubate overnight at 4°C. After incubation, remove the MAdCAM by aspirate, add 200 μl of PBS + 1% BSA, and block the plate at 37°C and 5% CO2 for 2 hours. During this incubation, prepare a dilution curve of the compound in 100% DMSO in a 96-well V-bottom plate. Then, transfer 1.75 μl of the diluted compound to a new 96-well U-bottom plate containing 20 μl of assay culture (phosphate-free DMEM + 25 mM HEPES + 1% BSA). Add an additional 155 μl of assay culture by mixing with a pipette. Incubate this mixture at 37°C and 5% CO2 for 15 minutes. After incubation, add 175 μl of assay culture medium containing 2e6 / mL RPMI8866 cells to the compound-containing wells without mixing, and incubate the plate at 37°C and 5% CO2 for a further 15 minutes. During this incubation, remove the MAdCAM-coated plate from the incubator and wash it twice with 200 μl PBS + 0.1% BSA. After incubating the cells with the compound for 15 minutes, pipette them out and mix, then transfer 100 μl of the mixture to the three-washed MAdCAM-coated plate. Next, incubate this plate at 37°C and 5% CO2 for 1 hour. After incubation, wash the plate twice with 200 μl phenol-free RPMI + 1% BSA and once with 50 μl. Add the final 50 μl of phenol-free RPMI + 1% BSA to the wells after the final wash. Next, 50 μl of Promega CellTiter-Glo is added to the wells. The plate is incubated in a shaker at 200 RPM for 2 minutes, followed by off-shaking for another 8 minutes, and then fluorescence is read using a Biotek Cytation 5 Plate Reader. The raw data is converted to inhibition % compared to the bottom of the curve and analyzed using a 4-parameter nonlinear curve in a prism to obtain IC. 50 and IC 90 To decide.

[0186] [Table 3] JPEG0007869261000076.jpg242163JPEG0007869261000077.jpg63162

[0187] [Table 4] JPEG0007869261000079.jpg232154JPEG0007869261000080.jpg228154JPEG00078692610 00081.jpg231155JPEG0007869261000082.jpg241154JPEG0007869261000083.jpg123154

[0188] Built-in by reference All U.S. patents and U.S. and PCT patent application publications referenced herein are incorporated herein by reference.

[0189] Equivalents Those skilled in the art will recognize or confirm, using conventional experimental methods, numerous equivalents to the specific embodiments of the present invention described herein. Such equivalents are intended to be encompassed within the following claims. The present invention encompasses the following aspects and embodiments. (Embodiment 1) A pharmaceutical composition comprising a compound selected from the group consisting of the following, or a pharmaceutically acceptable salt thereof. [ka] JPEG0007869261000085.jpg209129JPEG0007869261000086.jpg209129JPEG0007869261000087.jpg213129 (Embodiment 2) The compound, [ka] The pharmaceutical composition according to Embodiment 1, or a pharmaceutically acceptable salt thereof. (Embodiment 3) The compound,

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Claims

1. A method for producing 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid, comprising the following steps: a. Add tert-butoxide to a solution containing (methoxymethyl)triphenylphosphonium chloride, and then add 6-methoxy-4-(trifluoromethyl)nicotinaldehyde to provide (E)-2-methoxy-5-(2-methoxyvinyl)-4-(trifluoromethyl)pyridine. b. Adding TFA and water to (E)-2-methoxy-5-(2-methoxyvinyl)-4-(trifluoromethyl)pyridine and heating to provide 2-(6-methoxy-4-(trifluoromethyl)pyridine-3-yl)acetaldehyde, c. Azetidine hydrochloride is added to 2-(6-methoxy-4-(trifluoromethyl)pyridine-3-yl)acetaldehyde, and further NaBH(OAc) 3 The step of adding 5-(2-(azetidine-1-yl)ethyl)-2-methoxy-4-(trifluoromethyl)pyridine, d. A step of mixing 5-(2-(azetidine-1-yl)ethyl)-2-methoxy-4-(trifluoromethyl)pyridine with HBr / AcOH to provide 5-(2-(azetidine-1-yl)ethyl)-4-(trifluoromethyl)pyridine-2(1H)-one, e. Ethyl 4-methyl-2-(methylsulfonyloxy)pentanoate and K 2 CO 3 The steps include adding to 5-(2-(azetidine-1-yl)ethyl)-4-(trifluoromethyl)pyridine-2(1H)-one and heating to provide ethyl 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate, and f. Ethyl 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate is dissolved in ethanol and water with LiOH-H 2 The reaction mixture is treated with O in a molar ratio of approximately 1:5 to neutralize it and obtain 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid. The method, including the method described above.

2. The following steps: a. A step of mixing 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid with ethyl(3S)-3-amino-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate, TCFH, and NMI to provide ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate, or b. A step of mixing 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid with (S)-ethyl3-amino-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propanoate, TCFH, and NMI to provide (3S)-ethyl3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propanoate, or c. Mix 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid with ethyl(3S)-3-amino-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate, TCFH, and NMI. The step of providing ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate, or d. A step of mixing 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid with (S)-ethyl3-amino-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propanoate, TCFH, and NMI to provide (3S)-ethyl3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propanoate, or e. The step of mixing 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid with ethyl(3S)-3-amino-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate, TCFH, and NMI to provide ethyl(3S)-3-(2,(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate A manufacturing method that includes this.

3. The following steps: a. 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid is mixed with ethyl(3S)-3-amino-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate, TCFH, and NMI in acetonitrile. Mix in a molar ratio of approximately 0.3:0.3:0.45:1.5 to obtain ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propane The steps are: providing a noate, and treating ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoate with lithium hydroxide in ethanol and water to obtain (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propanoic acid, or b. 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid is mixed with (S)-ethyl 3-amino-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propanoate, TCFH, and NMI in acetonitrile at approximately 0.31:0.31: The steps of mixing in a molar ratio of 0.62:1 to provide (3S)-ethyl3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propanoate, and (3S)-ethyl3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4 The step of treating (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propanoic acid, or c. 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid is mixed with ethyl(3S)-3-amino-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate, TCFH, and NMI in acetonitrile at approximately 0.21:0.21: The steps include mixing in a molar ratio of 0.32:2.5 to provide ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate, and ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide) The step of treating (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoic acid with lithium hydroxide in methanol and water, or d. Mix 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid with (S)-ethyl3-amino-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propanoate, TCFH, and NMI in acetonitrile in a molar ratio of approximately 0.41:0.41:1.3:6.3 to obtain (3S)-ethyl3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propanoate The steps are: providing a noate, and treating (3S)-ethyl3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propanoate with lithium hydroxide in methanol and water to obtain (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propanoic acid, or e. 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid in acetonitrile with ethyl(3S)-3-amino-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate, TCFH, and NMI. Mix in a molar ratio of approximately 0.32:0.32:0.64:1.6 and ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl) The steps of providing propanoate and ethyl(3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoate, The step of treating with lithium hydroxide in water and ethanol to obtain (3S)-3-(2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanamide)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propanoic acid. The method according to claim 2, including the method described in claim 2.

4. The following steps: a. Add tert-butoxide to a solution of (methoxymethyl)triphenylphosphonium chloride, and then add 6-methoxy-4-(trifluoromethyl)nicotinaldehyde to provide (E)-2-methoxy-5-(2-methoxyvinyl)-4-(trifluoromethyl)pyridine; b. Add TFA and water to (E)-2-methoxy-5-(2-methoxyvinyl)-4-(trifluoromethyl)pyridine and heat to provide 2-(6-methoxy-4-(trifluoromethyl)pyridine-3-yl)acetaldehyde. c. Azetidine hydrochloride is added to 2-(6-methoxy-4-(trifluoromethyl)pyridine-3-yl)acetaldehyde, and further NaBH(OAc) 3 The step of adding 5-(2-(azetidine-1-yl)ethyl)-2-methoxy-4-(trifluoromethyl)pyridine, d. A step in which 5-(2-(azetidine-1-yl)ethyl)-2-methoxy-4-(trifluoromethyl)pyridine is mixed with HBr / AcOH and heated to provide 5-(2-(azetidine-1-yl)ethyl)-4-(trifluoromethyl)pyridine-2(1H)-one. e. Ethyl 4-methyl-2-(methylsulfonyloxy)pentanoate and K 2 CO 3 The steps include adding to 5-(2-(azetidine-1-yl)ethyl)-4-(trifluoromethyl)pyridine-2(1H)-one and heating to provide ethyl 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate, and f. Ethyl 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoate is LiOH-H 2 The method according to claim 2 or 3, comprising the step of treating the reaction mixture with O, ethanol, and water in a molar ratio of about 1:5 to neutralize the reaction mixture and obtain 2-(5-(2-(azetidine-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridine-1(2H)-yl)-4-methylpentanoic acid.

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