Inhibit human integrin A4B7

Abstract

Small molecule antagonists of human α4β7 integrin and methods for treating various diseases and symptoms using said small molecule antagonists are disclosed.

Description

[0001] Cross-referencing related applications

[0002] This application claims the priority of PCT International Application No. PCT / US20 / 55986, filed on October 16, 2020. Technical Field

[0003] Novel compounds and related methods for inhibiting α4β7 integrin are disclosed herein. The compounds and methods disclosed herein are applicable to the development of drugs for the treatment of α4β7 integrin-mediated conditions such as inflammatory bowel disease (IBD), ulcerative colitis (UC), and Crohn's disease (CD). Background Technology

[0004] Integrins are non-covalently associated α / β heterodimeric cell surface receptors involved in many cellular processes. Different expression of integrins can modulate cell adhesion properties, allowing for the recruitment of different leukocyte populations to specific organs in response to different inflammatory signals. α4-integrin, containing α4β7, plays a role in lymphocyte migration throughout the gastrointestinal tract. This integrin is expressed on most leukocytes, including B and T lymphocytes, where it mediates cell adhesion through selective binding to its primary ligand, mucosal addressing cell adhesion molecule 1 (MAdCAM). Memory T lymphocytes expressing α4β7 integrin preferentially migrate into the gastrointestinal tract via strong adhesion to mucosal vascular addressing cell adhesion molecule 1 (MAdCAM-1).

[0005] Inhibitors of specific integrin-ligand interactions have been used to treat a variety of diseases. For example, monoclonal antibodies exhibiting high binding affinity for α4β7 have shown therapeutic benefits for gastrointestinal autoinflammatory / autoimmune diseases such as Crohn's disease and ulcerative colitis. However, these therapies also have certain undesirable characteristics for patients. Monoclonal antibody α4β7 integrin inhibitors are administered via parenteral administration, have long half-lives, cannot be rapidly modified for exposure, and their activity is reduced due to the formation of anti-drug antibodies. Monoclonal antibody therapies can be challenging to manufacture compared to small molecule therapies. Furthermore, some therapies that inhibit α4β7 also interfere with α4β1 integrin-ligand interactions, resulting in dangerous side effects for patients. In immunosuppressed patients, activity at the α4β1 integrin level is involved in the development of progressive multifocal leukoencephalopathy (PML), a life-threatening and progressive brain infection.

[0006] There remains a medical need for effective and safe oral α4β7 integrin inhibitors with improved pharmacological properties, as an important complement to therapeutic medical devices for α4β7 integrin-mediated conditions such as inflammatory bowel disease (IBD), ulcerative colitis (UC), and Crohn's disease (CD). Summary of the Invention

[0007] This invention relates to compounds that antagonize α4β7 integrin and methods for preparing and isolating said compounds. Furthermore, the small molecule compounds disclosed herein exhibit favorable pharmaceutical properties, including oral bioavailability, ADME (absorption, distribution, metabolism, and excretion), pharmacokinetics, CYP inhibition, and / or other safety characteristics, which contribute to achieving therapeutic efficacy while minimizing undesirable properties. Attached Figure Description

[0008] Figure 1 This is a table summarizing the in vitro inhibition of α4β7 integrin by exemplary compounds (i.e., data obtained from fluorescence polarization assays in Example 4 and ligand binding assays in Example 5). Detailed Implementation

[0009] In some embodiments, the present invention relates to compounds that antagonize α4β7 integrin. These compounds could be used to treat diseases that can be treated by inhibiting α4β7 integrin (e.g., Crohn's disease (CD) and ulcerative colitis (UC)).

[0010] definition

[0011] For convenience, certain terms used in the specification, examples, and appended claims are collected herein before further description of the invention. These definitions should be understood in accordance with the remainder of this disclosure and as will be understood by those skilled in the art. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0012] To make the invention easier to understand, certain terms and phrases are defined below and throughout the specification.

[0013] The article “a and an” as used in this article refers to one or more (i.e., at least one) grammatical objects of the article. For example, “element” means one or more elements.

[0014] As used herein in the specification and claims, the phrase “and / or” should be understood to mean “any one or both” of the elements so combined, that is, elements that coexist in some cases and exist separately in others. Multiple elements listed with “and / or” should be understood in the same way, that is, “one or more” of the elements so combined. Other elements may optionally be present, whether related to or unrelated to those specifically identified by the “and / or” clause. Thus, as a non-limiting example, in one embodiment, when used in conjunction with open-ended language such as “comprising,” a reference to “A and / or B” may refer only to A (optionally including elements other than B); in another embodiment, only to B (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); and so on.

[0015] As used herein in this specification and claims, “or” should be understood to have the same meaning as “and / or” as defined above. For example, when items in a list are separated, “or” or “and / or” should be interpreted as inclusive, i.e., including a plurality of elements or at least one element in a list of elements, but also including more than one element and optionally other items not listed. Terms that clearly indicate the opposite, such as “only one of…” or “exact one of…” or, when used in claims, “consisting of…” will refer to including a plurality of elements or exactly one element in a list of elements. Generally, when preceded by an exclusive term such as “any one,” “one of…,” “only one of…,” or “exact one of…,” the term “or” as used herein should be interpreted only as indicating an exclusive alternative (i.e., “one or another, not two”). When used in claims, “consisting substantially of…” should have the ordinary meaning as used in the field of patent law.

[0016] As used herein in the specification and claims, the phrase "at least one" relating to a list having one or more elements should be understood to mean at least one element selected from any one or more elements in the element list, but not necessarily including at least one of every element specifically listed in the element list, and does not exclude any combination of elements in the element list. This definition also allows for the optional presence of elements other than those specifically identified in the element list referred to by the phrase "at least one," whether or not they are related to those specifically identified elements. Thus, as a non-limiting example, in one embodiment, "at least one of A and B" (or equivalently, "at least one of A or B," or equivalently, "at least one of A and / or B") may mean at least one element that optionally includes more than one A, has no B (and optionally includes elements other than B); in another embodiment, it may mean at least one element that optionally includes more than one B, has no A (and optionally includes elements other than A); in yet another embodiment, it may mean at least one element that optionally includes more than one A, and at least one element that optionally includes more than one B (and optionally includes other elements); and so on.

[0017] It should also be understood that, unless explicitly stated otherwise, in any method claimed herein that comprises more than one step or operation, the order of the steps or operations of the method is not necessarily limited to the order of the steps or operations of the method described herein.

[0018] In the claims and the foregoing description, all transitional phrases such as “comprising,” “including,” “carrying,” “having,” “containing,” “involving,” “accommodating,” “constituting,” “made of,” etc., should be understood as open-ended, meaning that they include but are not limited to. As described in Section 2111.03 of the Patent Examination Manual of the United States Patent and Trademark Office, only the transitional phrases “composed of” and “substantially composed of” should be closed or semi-closed transitional phrases, respectively.

[0019] Certain compounds contained in the compositions of the present invention may exist in specific geometric or stereoisomeric forms. Additionally, the polymers of the present invention may also be optically active. The present invention considers all such compounds, including cis and trans isomers, R- and S enantiomers, diastereomers, (D)-isomers, (L)-isomers, racemic mixtures thereof, and other mixtures thereof, to fall within the scope of the present invention. Additional asymmetric carbon atoms may be present in substituents (such as alkyl groups). All such isomers and mixtures thereof are intended to be included in the present invention.

[0020] For example, if a specific enantiomer of the compound of the present invention is desired, it can be prepared by asymmetric synthesis or by derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is isolated and the auxiliary groups are cleaved to provide the pure desired enantiomer. Alternatively, in the case where the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with a suitable optically active acid or base, and the resulting diastereomeric isomer is then reduced by stepwise crystallization or chromatographic methods well known in the art, and the pure enantiomer is subsequently recovered.

[0021] The structures described herein also refer to compounds that differ only in the presence of one or more isotopically enriched atoms. For example, by substituting hydrogen or deuterium with deuterium or tritium. 13 C- or 14 Compounds produced by C-enrichment of carbon to replace carbon are within the scope of this invention.

[0022] As used in this article, the terms “α4β7”, “a4B7”, “a4b7”, “α-4β-7”, and “α4β7” all refer to α4β7.

[0023] As used herein, the phrase "pharmaceutically acceptable excipient" or "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or medium, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulation material, that carries or transports the subject chemical from one organ or part of the body to another organ or part of the body. Each carrier must be "acceptable," harmless to the patient, and substantially pyrogen-free in the sense of compatibility with the other components of the formulation. Examples of materials that can serve as pharmaceutically acceptable carriers include: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch and potato starch; (3) cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose, and cellulose acetate; (4) powdered tragacanth gum; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter; (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; (10) diols such as propylene glycol; (11) polyols such as glycerol, sorbitol, mannitol, and polyethylene glycol; (12) esters such as ethyl oleate and ethyl laurate; (13) agar; (14) buffers such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; and (18) Ringer's solution. (19) ethanol; (20) phosphate buffer solution; and (21) other non-toxic and compatible substances used in the pharmaceutical formulation. In some embodiments, the pharmaceutical compositions of the present invention are pyrogen-free when administered to a patient, i.e., do not induce a significant increase in temperature.

[0024] The term "pharmaceutically acceptable salt" refers to a relatively non-toxic inorganic and organic acid addition salt of one or more compounds. These salts can be prepared in situ during the final isolation and purification of one or more compounds, or by reacting one or more purified compounds, in their free base form, alone with a suitable organic or inorganic acid and isolating the resulting salt. Representative salts include hydrobromide, hydrochloride, sulfate, hydrogen sulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, lysinate, benzoate, lactate, phosphate, toluenesulfonate, citrate, maleate, fumarate, succinate, tartrate, naphthate, methanesulfonate, glucono-p-ethyl, lacturonate, and lauryl sulfonate, etc. (See, for example, Batelvi et al., (1977), "Pharmaceutical Salts", J. Pharm. Sci. 66:1-19).

[0025] In other cases, compounds that can be used in the methods of the present invention may contain one or more acidic functional groups and are therefore capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable bases. In these cases, the term "pharmaceutically acceptable salt" refers to a relatively non-toxic inorganic and organic base addition salt of one or more compounds. These salts can also be prepared in situ during the final separation and purification of one or more compounds, or by reacting one or more purified compounds in their free acid form with a suitable base (such as a hydroxide, carbonate, or bicarbonate of a pharmaceutically acceptable metal cation), with ammonia, or alone with a pharmaceutically acceptable primary, secondary, or tertiary organic amine. Representative bases or alkaline earth metal salts include lithium, sodium, potassium, calcium, magnesium, and aluminum salts, etc. Representative organic amines that can be used to form base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine, etc. (see, for example, Berge et al., ibid.).

[0026] The "therapeutic effective amount" (or "effective amount") of a compound used in treatment refers to the amount of the compound in a formulation that, when administered as part of a desired dosing regimen (to a mammal, preferably a human), relieves symptoms, improves condition, or slows the onset of disease symptoms for the purpose of treating the condition or symptom or for cosmetic purposes, according to clinically acceptable criteria (e.g., a reasonable benefit / risk ratio applicable to any medical treatment).

[0027] The term "preventive or therapeutic" treatment is recognized in the art and includes the administration of one or more of the subject compositions to a host. A treatment is preventive (i.e., it protects the host from developing the undesirable condition) if it is administered before the clinical manifestation of an undesirable symptom (e.g., disease or other undesirable condition in the host animal), and therapeutic (i.e., it aims to alleviate, improve, or stabilize the existing undesirable symptom or its side effects) if it is administered after the manifestation of the undesirable symptom.

[0028] The term "patient" refers to a mammal that requires specific treatment. In some embodiments, the patient is a primate, dog, cat, or horse. In some embodiments, the patient is a human.

[0029] As used herein, the term "prodrug" encompasses compounds that are converted into therapeutically active agents under physiological conditions. A common method for preparing prodrugs involves hydrolyzing selected portions of the molecule under physiological conditions to reveal the desired molecule. In other embodiments, prodrugs are converted via enzymatic activity in a host animal.

[0030] For the purposes of this invention, chemical elements are identified according to the Periodic Table of the Elements, CAS edition, Handbook of Chemistry and Physics, 67th edition, 1986-87, inner pages.

[0031] Exemplary compounds

[0032] In some embodiments, the present invention relates to compounds selected from the group consisting of:

[0033]

[0034]

[0035]

[0036] In some embodiments, the present invention relates to any of the compounds mentioned above, wherein the compounds are in the form of pharmaceutically acceptable salts.

[0037] In some embodiments, the present invention relates to compounds selected from the group consisting of:

[0038]

[0039]

[0040]

[0041]

[0042]

[0043] Pharmaceutical Composition

[0044] The compounds disclosed herein can be formulated into a variety of pharmaceutical compositions. The compounds disclosed herein, and their pharmaceutically acceptable salts, can be active pharmaceutical ingredients (APIs) that are combined with one or more other components to form a pharmaceutical substance (DS) pharmaceutical composition. A pharmaceutical substance (DS) pharmaceutical composition may include an API (i.e., the compound disclosed herein or its pharmaceutically acceptable salt) and one or more pharmaceutically acceptable carriers, diluents, and / or excipients. The carrier, diluent, or excipient may be selected to be compatible with other components of the formulation and to be appropriately safe and effective for the intended therapy. The desired weight concentration of the compound as an active pharmaceutical ingredient (API) may be combined with other inactive ingredients to form a pharmaceutical substance (DS) in a formulation batch. Pharmaceutically acceptable compositions can be formulated for administration via an appropriate route, such as oral delivery in unit dosage forms (including capsules or tablets). Such compositions can be prepared by associating an active pharmaceutical ingredient (API) comprising a compound of formula (I) with a carrier or excipient.

[0045] In some embodiments, the present invention provides a pharmaceutical composition formulated for 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 oral therapeutic administration of the α4β7 integrin inhibitor compound.

[0046] In some embodiments, the present invention provides a pharmaceutical composition comprising the compound or a pharmaceutically acceptable salt thereof as an active pharmaceutical ingredient (API).

[0047] In some embodiments, the present invention relates to a pharmaceutical composition comprising a compound selected from the group consisting of, or a pharmaceutically acceptable salt thereof, as an active pharmaceutical ingredient (API):

[0048]

[0049]

[0050]

[0051]

[0052] Pharmaceutically acceptable compositions including the compounds of the present invention can be prepared by various procedures. For example, the compounds can be formulated with suitable excipients, diluents or carriers and formed into tablets or capsules and other suitable dosage forms.

[0053] Pharmaceutical compositions may be provided in unit doses containing a predetermined amount of an API, including the compounds / unit doses of the present invention. 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. Therefore, such unit doses may be administered at desired dosing intervals. The concentration of the active compound in the pharmaceutical composition will depend on various applicable parameters and considerations, such as the absorption rate, inactivation rate, and excretion rate of the drug, as well as other factors known to those skilled in the art. It should be noted that the dosage values ​​will also vary with the severity of the condition to be alleviated. It should be further understood that, for any particular subject, the specific dosing 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 the concentration ranges set forth herein are merely exemplary and not intended to limit the scope and practice of the claimed compositions. The active ingredient may be administered once or divided into many smaller doses administered at different time intervals.

[0054] In some embodiments, the active compound is administered orally. Oral compositions will typically contain an inert diluent or an edible carrier. They may be encapsulated in gelatin capsules or compressed into tablets. For oral therapeutic administration, the active compound may be incorporated together with excipients and used in tablet, lozenge, or capsule form. Pharmaceutically compatible binder and / or adjuvant materials may be included as part of the composition. Pharmaceutical compositions comprising compounds of the present invention formulated for oral delivery may be prepared in unit dosage forms, such as capsules at the desired dose strength of the compound. For oral administration in liquid form, the oral pharmaceutical component may be combined with any orally administered, non-toxic, pharmaceutically acceptable inert carrier, such as ethanol, glycerin, water, etc. For oral administration in tablet or capsule form, the compound may be combined with an orally administered, non-toxic, pharmaceutically acceptable inert carrier. Other examples of excipients, diluents, and carriers suitable for such formulations include: fillers and extenders such as starch and sugar; and binders such as cellulose derivatives. In addition, suitable binders, lubricants, disintegrants, and colorants may be incorporated into the mixture when desired or required. Suitable binders include starch, natural sugars, natural and synthetic gums, etc. Lubricants and / or flow aids may be used in these dosage forms.

[0055] Tablets, pills, capsules, lozenges, etc., may contain any of the following ingredients or compounds with similar properties: binders such as microcrystalline cellulose, tragacanth gum, or gelatin; excipients such as starch or lactose; disintegrants such as alginic acid, Primogel, or corn starch; lubricants such as magnesium stearate or sterotes; glidants such as colloidal silica; sweeteners such as sucrose or saccharin; or flavorings such as peppermint, methyl salicylate, or orange flavorings. When the dosage unit is in capsule form, in addition to the above types of materials, it may also contain a liquid carrier such as fatty oil. Furthermore, the dosage unit form may contain various other materials that alter the physical form of the dosage unit, such as coatings of sugar or other enteric solvents.

[0056] The compound can be applied as a component of elixirs, suspensions, syrups, flakes, etc. In addition to one or more active compounds, syrups may also contain sucrose or sweeteners as sweeteners, as well as certain preservatives, dyes and colorants, and flavorings.

[0057] The compounds can be formulated into solutions suitable for parenteral administration, such as via intramuscular, subcutaneous, or intravenous routes. For example, the compounds of the present invention can be dissolved in a suitable buffer solution. Pharmaceutical compositions comprising the compounds at desired concentrations can be formulated into injectable drug solutions (usefully, for example, in preclinical animal studies).

[0058] Exemplary methods

[0059] Compounds that inhibit α4β7 could be used to develop drugs for treating patients with ulcerative colitis and Crohn's disease. Patients with ulcerative colitis (UC) and Crohn's disease (CD) suffer from autoimmune inflammation of the digestive tract, and for many of these patients, CD4... + Memory T cells drive disease progression and onset through their ability to secrete pro-inflammatory and effector cytokines within the gut, thereby influencing peripheral immune cells and tissues. The progression and onset of these disease symptoms are believed to involve T cell extravasation, allowing blood to enter intestinal tissues and leading to inflammatory symptoms identified in UC and CD via integrin-associated mechanisms. Inhibition of α4β7 can disrupt this mechanism, preventing T cells from localizing to tissues and effectively treating and preventing diseases such as UC and CD. Gut-associated T cells require surface expression of integrin α4β7 and the chemokine receptor CCR9. While CCR9 is used by cells for gradient migration against CCL25 expressed in the small intestine, α4β7 is a binding ligand for the mucosal addressin cell adhesion molecule 1 (MAdCAM-1). Integrin α4β7 binds to MAdCAM-1 with high affinity, thereby promoting cell rolling and firm adhesion, followed by extravasation into tissues.

[0060] The pharmaceutical composition may include a compound that inhibits α4β7 integrin on inflammatory cells, the compound enabling these cells to adhere to mucosal addressin cell adhesion molecule-1 (MAdCAM-1) and inhibiting or preventing these cells from entering the lamina propria and gut-associated lymphoid tissue.

[0061] The compounds of the present invention were evaluated using fluorescence polarization (FP) assays, as described in Example 4. FP assays were used to evaluate the potency of the compounds for purified proteins. FP assays consist of measuring the extracellular domain or headpiece of the purified integrin αβ-dihexadimer bound to substituted or truncated ligands. Results of FP assays for exemplary compounds are provided herein.

[0062] The compounds of the present invention were further evaluated using a ligand binding assay (LBA) as described in Example 5 to examine the potency of the free ligands in binding to receptors expressed on cells. The MAdCAM ligand binding assay measured the binding of fluorescently labeled MAdCAM-1-Fc to RPMI 8866 cells in the presence of Mn++ using flow cytometry. This assay assesses the binding of the compounds to the native full-length receptors on the cell surface. One advantage of the MAdCAM ligand binding assay is its ability to quantify and distinguish the activity of potent compounds exceeding the functional sensitivity limit of the FP assay [Mn ~ 10 nM]. The ligand binding assay (LBA) was used to examine the potency of the compounds and the selectivity of the free ligands in binding to receptors expressed on cells.

[0063] In some embodiments, the present invention relates to any of the methods mentioned above, wherein the subject is a mammal. In some embodiments, the present invention relates to any of the methods mentioned above, wherein the subject is a human.

[0064] Example

[0065] The invention described herein in general will be more readily understood by referring to the following examples, which are included only for the purpose of illustrating certain aspects and embodiments of the invention and are not intended to limit the invention.

[0066] Examples 1 to 3 describe the synthesis of the compounds of the present invention. These compounds can be prepared as mixtures of diastereomers having a (3S) configuration (i.e., with the carboxylic acid moiety at the stereocenter β) and mixtures of diastereomers covalently bonded to the nitrogen atom of the pyridone ring of the compound at the chiral center.

[0067] The examples show compounds with significant activity in the fluorescence polarization (FP) assay of Example 4, along with specific diastereomers. Example 4 describes the fluorescence polarization (FP) assay. Example 5 describes the ligand binding (LB) assay. Example 6 describes the cell adhesion (CA) assay.

[0068] Example 1. A general scheme for synthesizing α4β7 inhibitors

[0069] β-amino acid synthesis

[0070] The synthesis of β-amino acids can be achieved using well-known procedures described in the literature, such as, but not limited to, “Enantioselective Synthesis of β-Amino Acids,” 2nd ed., edited by Eusebio Juaristi and Vadim A. Soloshonok, first published January 27, 2005, John Wiley & Sons, Inc.; Ellman et al., *Acc. Chem. Res.*, 2002, 35, 984-995; Franklin A. Davis and Bang-Chi Chen, *Chem. Soc. Rev.*, 1998, 27, 13-18; Jacobsen, MF; Skrydstrup, T., *J. Org. Chem.*, 2003, 68, 7122; Tang, TP; Ellman, JA, *J. Org. Chem.*, 2002, 67, 7819; and Tang, TP; Ellman, JA, *J. Org. Chem.*, 1999, 64, 12.

[0071] Reductive amination

[0072]

[0073] Procedure A: A mixture of amine (1 equivalent) and aldehyde (1.2 equivalent) in DCM (1–2 mL / mmol amine) was stirred at room temperature for 30 minutes. Then, NaBH(OAc)3 (1.5 equivalent) was added in portions, 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 provide the desired amine.

[0074] Procedure B: A mixture of aldehyde (1 equivalent), amine (1.05–2 equivalents) in DCE (3–4 mL / mmol aldehyde) is stirred at room temperature for 10–30 minutes. Then, NaBH(OAc)₃ (3–4 equivalents) is added in portions, and the mixture is stirred at room temperature for 1–16 minutes until the reaction is complete by LC / MS. The solvent is concentrated under vacuum, and the residue is purified by silica gel chromatography to provide the desired amine.

[0075] Procedure C: Aldehyde (1 equivalent), AcOH (1.2 equivalent), and amine (1.05–2 equivalents) were added to a mixture of DCM (2–3 mL / mmol aldehyde) and MeOH (0.5 mL / mmol aldehyde) and stirred at room temperature for 15–30 minutes. Then, NaBH(OAc)₃ (2 equivalents) was added in portions, and the mixture was stirred at room temperature for 1–16 minutes until the reaction was complete by LC / MS. The solvent was concentrated under vacuum, and the residue was purified by silica gel chromatography to provide the desired amine.

[0076] Alkylation

[0077]

[0078] Procedure A: Add methanesulfonate (1.5 equivalents) and K₂CO₃ (3 equivalents) to a solution of amine (1 equivalent) in MeCN (3-4 mL / mmol amine). Stir the reaction mixture at 80 °C for 16 hours. Concentrate the reaction mixture under vacuum and purify the residue by reversed-phase HPLC to obtain the desired alkylated product.

[0079]

[0080] Procedure B: Add alkyl bromide (2 equivalents) and K₂CO₃ (2 equivalents) to a solution of amine (1 equivalent) in MeCN (3-4 mL / mmol amine). Stir the reaction mixture at 80 °C for 16 hours. Concentrate the reaction mixture under vacuum and purify the residue by reversed-phase HPLC to obtain the desired alkylated product.

[0081] Phenol deprotection

[0082]

[0083] The mixture of methoxypyridine (1 equivalent) in 44% HBr / AcOH (10 mL / mmol substrate) was heated at 55–75 °C for 5–16 hours until complete by LC-MS. The reaction was concentrated under vacuum, and the residue was purified by reversed-phase HPLC to give the desired phenolated product.

[0084] Wittig Reaction

[0085]

[0086] Procedure A: A mixture of (methoxymethyl)triphenylphosphine chloride (1.5 equivalents), t-BuOK (2.5 equivalents), and dioxane (2 mL / mmol phosphate) was stirred at room temperature for 15 minutes. Then, THF (1 mL / mmol aldehyde) containing an aldehyde (1 equivalent) was added. The mixture was stirred at room temperature for 2–16 hours. The reaction mixture was treated (diluted with water and extracted with EtOAc; the extracts were combined, dried over Na₂SO₄, filtered, and concentrated), and purified by silica gel chromatography to obtain the enol ether product.

[0087] Procedure B: A mixture of (methoxymethyl)triphenylphosphine chloride (1.1 equivalents), t-BuOK (2.5 equivalents), and THF (4 mL / mmol phosphate) was stirred at 0 °C for 1 hour. Then, THF containing an aldehyde (1 equivalent) (2 mL / mmol aldehyde) was added. The mixture was stirred at room temperature for 16 hours. The reaction mixture was treated (diluted with water and extracted with EtOAc; the extracts were combined, dried over Na₂SO₄, filtered, and concentrated), and purified by silica gel chromatography to obtain the enol ether product.

[0088] Enol ethers to aldehydes

[0089]

[0090] Procedure A: Treat the enol ether (1 equivalent) with TFA (2 mL / mmol) at room temperature for 4 hours. Remove the solvent under vacuum to provide the desired aldehyde.

[0091] Procedure B: Treat the enol ether (1 equivalent) with HCOOH (2 mL / mmol) at 70 °C for 2 hours. Remove the solvent under vacuum to provide the desired aldehyde.

[0092] Procedure C: Add TFA (2 mL / mmol) and water (0.25 mL / mmol) to a solution of enol ether (1 equivalent) in DCM (15 mL / mmol enol ether). Stir the reaction at 45 °C for 18 hours. Treat the reaction (quench with NaHCO3, extract with DCM; combine the extracts, dry with Na2SO4, filter and concentrate) to produce the desired aldehyde.

[0093] Stille reaction

[0094]

[0095] Pd(PPh3)4 (0.1 equivalent) was added to a solution of aryl bromide (1 equivalent) and allyl bromide (1.2 equivalent) in DMF (3 mL / mmol aryl bromide) under N2 conditions. The reaction was stirred at 100 °C for 16 h. The reaction was concentrated under vacuum, diluted with EtOAc, poured into a 20% KF aqueous solution, stirred for 1 h, and extracted. The combined organic layers were subjected to Na2SO4. 4干燥， The product is filtered and concentrated, and purified by silica gel chromatography to provide the desired product.

[0096] Olefin to Aldehyde

[0097]

[0098] K₂O₄⁻ 2H₂O (0.01 equivalents) was added to a solution of olefin (1 equivalent) in THF / H₂O (1:1) (10 mL / mmol olefin) at 0 °C. The mixture was stirred at 0 °C for 5 min, and then H₂O (1 mL / mmol olefin) containing NaIO₄ (3 equivalents) was added dropwise. The mixture was stirred at 0 °C for 1 h, then warmed to room temperature and stirred until complete by LC-MS. The reaction was then treated (diluted with water and extracted with EtOAc; the combined organic layers were dried over Na₂SO₄, filtered, and concentrated) to obtain the desired aldehyde.

[0099] Ester to acid

[0100]

[0101] The ester (1 equivalent) was treated with MeOH (1-3 mL / mmol ester) containing LiOH-H₂O (3-5 equivalents) and water (1-3 mL / mmol ester) at room temperature for 1-5 hours. The reaction was acidified to pH 3 with 1N HCl and concentrated. The residue was purified by preparative HPLC to give the desired carboxylic acid product.

[0102] Amine protection

[0103]

[0104] A mixture of amine (1 equivalent), DIEA (3 equivalents), and Boc₂O (2 equivalents) was stirred in DCM (5 mL / mmol) at room temperature for 16 hours until complete as measured by LC-MS. The reaction was then treated (washed with 0.5 N HCl, saturated NaHCO₃, and brine; extracted with DCM; the combined organic layers were dried over Na₂SO₄, filtered, and concentrated), and purified by silica gel chromatography.

[0105] Preparation of arylboranes

[0106]

[0107] A mixture of aryl bromide (1 equivalent), B2pin2 (1.2 equivalent), Pd(dppf)Cl2 (0.05 equivalent), and KOAc (3 equivalent) in dioxane (10 mL / mmol aryl bromide) was stirred at 110 °C for 2–5 hours under N2 until complete by LCMS. The reaction was filtered, concentrated under vacuum, and purified by silica gel chromatography to provide the desired arylborane.

[0108] Suzuki Couplets

[0109] "Palladium-catalyzed cross-coupling reactions of organoboron compounds" A. Suzuki, Chem. Rev., 1995, 957, 2457-2483.

[0110]

[0111] Procedure A: An aryl bromide (1.2 equivalents), Pd(dppf)Cl2 (0.1 equivalents), K2CO3 (2 equivalents), and water (2 mL / mmol) were added to a solution of arylborane (1 equivalent) in dioxane (10 mL / mmol arylborane). The reaction was stirred at 110 °C for 3 hours under N2. The reaction was then treated (washed with brine and extracted with EtOAc; the extracts were combined, dried over Na2SO4, filtered, and concentrated) and purified by silica gel chromatography to yield the desired diaryl product.

[0112]

[0113] Procedure B: A solution of aryl bromide (1 equivalent) and arylborane (1.1 equivalent) in dioxane (10 mL / mmol aryl bromide) was added to water (2 mL / mmol) containing K₂CO₃ (2 equivalents) and Pd(dppf)Cl₂ (0.1 equivalent). The reaction was stirred at 110 °C for 2 hours under N₂. The reaction was then treated (washed with brine and extracted with EtOAc; the extracts were combined, dried over Na₂SO₄, filtered, and concentrated) and purified by silica gel chromatography to yield the desired diaryl product.

[0114]

[0115] Procedure C: A mixture of aryl bromide (1 equivalent), arylborane (2.0 equivalent), K₂CO₃ (3 equivalent), and Pd(dppf)Cl₂ (0.05 equivalent) in dioxane (10 mL / mmol aryl bromide) and water (1 mL / mmol) was stirred at 110 °C for 2 hours under N₂ until complete by LC-MS. The reaction was treated (washed with brine and extracted with EtOAc; the extracts were combined, dried over Na₂SO₄, filtered, and concentrated) and purified by silica gel chromatography to yield the desired diaryl product.

[0116] Boc deprotection

[0117]

[0118] Add 4M HCl-dioxane (12 equivalents) to DCM (4 mL / mmol amine) containing a Boc-protected amine (1 equivalent). Stir the reaction for 1–2 hours until complete by LC-MS. Concentrate the reaction under vacuum to obtain the desired amine.

[0119] tert-butylsulfinyl deprotection

[0120]

[0121] Add 4M HCl-dioxane (1.7 equivalents) to a solution of tert-butylsulfinylamine (1 equivalent) in DCM (0.5 mL / mmol amine). Stir the reaction for 0.5–1 h until complete by LC-MS. Concentrate the reaction mixture and purify it by preparative HPLC to obtain the desired amine.

[0122] amide bond formation

[0123] "Peptide Coupling Reagents, More Than a Letter Soup" A. El-Faham, F. Albericio, *Chemical Review*, 2011, 111, 11, 6557-6602; "Amide Bond Formation and Peptide Coupling" CAGN Montalbetti, V. Falque, *Tetrahedron*, 2005, 61, 10827-10852.

[0124]

[0125] 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 complete by LCMS. The reaction was concentrated under vacuum and purified by silica gel chromatography to obtain the desired amide product.

[0126] Ester hydrolysis

[0127]

[0128] The ester (1 equivalent) was treated with MeOH (1-3 mL / mmol ester) containing LiOH-H₂O (3-5 equivalents) and water (1-3 mL / mmol ester) at room temperature for 1-5 hours. The reaction 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.

[0129] Analytical methods

[0130] LCMS analysis method

[0131] The final compounds were analyzed using LC / MS conditions with UV detectors at 214 nm and 254 nm, and mass spectrometry scans of 110–800 amu in ESI+ ionization mode.

[0132] LC / MS A: Column: XBridge C18, 4.6 x 50 mm, 3.5 μm; Mobile phase: Water (10 mM ammonium bicarbonate), BCH3CN; Gradient: 5% - 95% B, 1.4 min, then hold for 1.6 min; Flow rate: 1.8 mL / min; Oven temperature: 50 °C.

[0133] LC / MS B: Column: SunFire C18, 4.6 x 50 mm, 3.5 μm; Mobile phase: Water (0.01% TFA), B CH3CN; Gradient: 5%-95% B, 1.5 min, then hold for 1.5 min; Flow rate: 2.0 mL / min; Oven temperature: 50 °C.

[0134] LC / MS C: Column: XBridge C18, 4.6 x 50 mm, 3.5 μm; Mobile phase: Water (10 mM ammonium bicarbonate), BCH3CN; Gradient: 5% - 95% B, 1.5 min, then hold for 1.5 min; Flow rate: 1.8 mL / min; Oven temperature: 50 °C.

[0135] LC / MS D: Column: Poroshell 120EC-C138, 4.6 x 30 mm, 2.7 μm; Mobile phase: Water (0.01% TFA), B CH3CN (0.01% TFA); Gradient: 5%-95% B, 1.2 min, then hold for 1.8 min; Flow rate: 2.2 mL / min; Oven temperature: 50 °C.

[0136] Example 2A. Preparation of intermediates

[0137] Preparation of ethyl (3S)-3-amino-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate

[0138] Step 1: 2,6-Dibromo-4-fluoro-3-methylaniline

[0139]

[0140] Bromine (52 mL, 1.0 mol) was added dropwise over 1.5 hours to a mixture of 4-fluoro-3-methylaniline (50.0 g, 400 mmol) in MeOH (120 mL) and DCM (120 mL) at room temperature, and the mixture was stirred for 4 hours at room temperature. 1N Na₂S₂O₃ aqueous solution (300 mL) and ethyl acetate (500 mL) were added, and the mixture was stirred for 10 minutes, followed by alkalization 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 brine (200 mL), dried over Na₂SO₄, filtered, and concentrated under vacuum to provide 2,6-dibromo-4-fluoro-3-methylaniline (80 g) as a white solid. Yield 70.7% (ESI 284.0 [M+H]). + ).

[0141] Step 2: 4-Fluoro-2,3,6-Trimethylaniline

[0142]

[0143] 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) was mixed with methylboronic acid (49.0 g, 819 mmol), K₂CO₃ (111.0 g, 819 mmol), and a 1,1'-bis(diphenylphosphino)ferrocene-dichloropalladium(II)dichloromethane complex (10.0 g, 13.65 mmol). The mixture was stirred overnight at 110 °C. The reaction mixture was poured into water (500 mL) and extracted with ethyl acetate (500 mL). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (petroleum ether: EtOAc 1:5) to yield 4-fluoro-2,3,6-trimethylaniline (20.0 g) as a colorless oil. Yield 47.6% (ESI 154.3(M+H)) + )

[0144] Step 3: 2-Bromo-5-fluoro-1,3,4-trimethylbenzene

[0145]

[0146] t-BuONO (3.8 g, 37.2 mmol) was added to a mixture of 4-fluoro-2,3,6-trimethylaniline (3.8 g, 24.8 mmol) and MeCN (30 mL) 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 chromatography (petroleum ether) to provide 2-bromo-5-fluoro-1,3,4-trimethylbenzene (1.3 g) as a colorless oil. Yield: 33.9%.

[0147] 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)propionate

[0148]

[0149] 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-Phos Pd G2 (285 mg, 0.33 mmol) in dioxane (10 mL) and H2O (2 mL) was stirred at 110 °C for 2 hours under a nitrogen atmosphere. The mixture was poured into water (100 mL) and extracted with EtOAc (100 mL x 3). The combined organic layers were dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography (petroleum ether:EtOAc 1:1) to yield ethyl (3S)-3-(((R)-tert-butylsulfinyl)amino)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (380 mg), a yellow oil. Yield 21.3% (ESI 538.0 (M+H)). + )

[0150] Step 5: Ethyl (3S)-3-amino-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate

[0151]

[0152] Ethyl (3S)-3-(((R)-tert-butylsulfinyl)amino)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (380 mg, 0.70 mmol) in DCM (5 mL) was added to HCl-dioxane (4 M, 5 mL) and 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)propionate (340 mg) as a yellow oil, which was used directly in the next step without further purification. Yield 100% (ESI 434.2 [M+H) + ).

[0153] Preparation of ethyl (S)-3-amino-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethyl-[1,1'-biphenyl]-3-yl)propionate

[0154] Step 1: (S)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)-3-((R)-1,1-dimethylethylsulfinamide)propionate

[0155]

[0156] X-Phos Pd G2 (79 mg, 0.1 mmol) was added to a mixture of (S)-3-(3-bromo-5-cyclopropyl-2,6-difluorophenyl)-3-((R)-1,1-dimethylethylsulfinamide)propionate (600 mg, 1.3 mmol), trimethylphenylboronic 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 to 110 °C under a nitrogen atmosphere for 2 hours. Water (20 mL) was added, and the solution was extracted with EtOAc (20 mL x 3). The combined organic phases were concentrated under vacuum, and the residue was purified by silica gel column chromatography (petroleum ether: EtOAc 2:1) to give ethyl (S)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)-3-((R)-1,1-dimethylethylsulfinamido)propionate (400 mg) as a dark solid. Yield 80% (ESI 492.1 [M-100+H] + ).

[0157] Step 2: Ethyl (S)-3-amino-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethyl-[1,1'-biphenyl]-3-yl)propionate

[0158]

[0159] Ethyl (S)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)-3-((R)-1,1-dimethylethylsulfinamido)propionate (550 mg, 1.12 mmol) in a mixture of DCM (1 mL) and EtOH (2 mL) was added with 4 M HCl-dioxane (2 mL, 5.0 mmol), and the mixture was stirred at room temperature for 0.5 h. The mixture was then concentrated under vacuum to produce ethyl (S)-3-amino-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethyl-[1,1'-biphenyl]-3-yl)propionate (360 mg, crude product) as a yellow oil, which was used directly in the next reaction. (ESI 388.1(M+H) + ).

[0160] Preparation of ethyl (3S)-3-amino-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propionate

[0161] Step 1: Ethyl (S)-3-(((R)-tert-butylsulfinyl)amino)-3-(3-cyclopropyl-2,6-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxacyclopentaborane-2-yl)phenyl)propionate

[0162]

[0163] A mixture of (S)-3-(3-bromo-5-cyclopropyl-2,6-difluorophenyl)-3-(((R)-tert-butylsulfinyl)amino)propionate (800 mg, 1.8 mmol), KOAc (529 mg, 5.4 mmol), Xphos-PdG2 (142 mg, 0.18 mmol), and 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bis(1,3,2-dioxacyclopentaborane) (686 mg, 2.7 mmol) in dioxane (10 mL) was stirred at 110 °C for 2 hours under nitrogen atmosphere. The reaction mixture was poured into 15 mL of water and extracted with EtOAc (15 x 3 mL x 3). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (petroleum ether:EtOAc 3:1) to yield ethyl (S)-3-(((R)-tert-butylsulfinyl)amino)-3-(3-cyclopropyl-2,6-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaneborane-2-yl)phenyl)propionate (500 mg) as a colorless oil. Yield 56% (ESI 500.2(M+H)). + )

[0164] Step 2: Ethyl (3S)-3-(((R)-tert-butylsulfinyl)amino)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propionate

[0165]

[0166] Ethyl (S)-3-(((R)-tert-butylsulfinyl)amino)-3-(3-cyclopropyl-2,6-difluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)phenyl)propionate (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) were stirred at 110 °C for 2 hours under a nitrogen atmosphere. The reaction mixture was poured into 15 mL of water and extracted with EtOAc (15 x 3 mL x 3). The combined organic layers were washed with brine (5 mL), dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under vacuum. The residue was purified by silica gel column chromatography (petroleum ether:EtOAc 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)propionate (200 mg) as a colorless oil. Yield 40% (ESI 496.2 (M+H)) + )

[0167] Step 3: Preparation of ethyl (3S)-3-amino-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propionate

[0168]

[0169] To 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)propionate (200 mg, 0.4 mmol) in EtOH (2 mL), HCl-dioxane (4 M, 2 mL, 8.0 mmol) was added. The mixture was stirred at room temperature for 1 hour. The mixture was concentrated under vacuum, and the residue was purified by reverse HPLC on a C18 / 40 g column (A: water / 0.01% TFA, B: MeOH, 0–100%) to provide ethyl (3S)-3-amino-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propionate oil (120 mg). Yield 76% (ESI 392.2 [M+H]) + ).

[0170] Example 2B. Preparation of intermediates

[0171] Preparation of 2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoic acid

[0172] Step 1: (E)-2-methoxy-5-(2-methoxyvinyl)-4-(trifluoromethyl)pyridine

[0173]

[0174] Potassium tert-butoxide (376 mg, 3.35 mmol) was added to a solution of (methoxymethyl)triphenylphosphine chloride (1.0 g, 2.95 mmol) in THF (13.406 mL) at 0 °C. After stirring at 0 °C for 1 hour, 6-methoxy-4-(trifluoromethyl)nicotinaldehyde (550 mg, 2.68 mmol) was added to a solution of THF (6.5 mL). The reaction was stirred overnight at room temperature and quenched with NH4Cl solution. The mixture was extracted (EtOAc x 3), concentrated, and purified by silica gel chromatography (0-100 ethyl acetate:hexane) to yield (E)-2-methoxy-5-(2-methoxyethylene)-4-(trifluoromethyl)pyridine (450 mg). Yield 72% (ESI 234.2 (M+H)). + ).

[0175] Step 2: 2-(6-methoxy-4-(trifluoromethyl)pyridin-3-yl)acetaldehyde

[0176]

[0177] TFA (0.595 mL, 7.72 mmol) and water (0.591 mL, 32.8 mmol) were added to a solution of (E)-2-methoxy-5-(2-methoxyvinyl)-4-(trifluoromethyl)pyridine (450 mg, 1.930 mmol) in DCM (29.689 mL). The reaction was stirred at 45 °C for 18 h. The reaction was diluted with DCM and quenched with NaHCO3. The mixture was washed with water, dried over Na2SO4, filtered, and concentrated to provide 2-(6-methoxy-4-(trifluoromethyl)pyridin-3-yl)acetaldehyde (343 mg), which was used without further purification. Yield 81% (ESI 220.18(M+H)). + ).

[0178] Step 3: 5-(2-(azacyclobutane-1-yl)ethyl)-2-methoxy-4-(trifluoromethyl)pyridine

[0179]

[0180] Azacyclobutane hydrochloride (3.4 g, 36.2 mmol) was added to a solution of 2-(6-methoxy-4-(trifluoromethyl)pyridin-3-yl)acetaldehyde (4 g, 18.1 mmol) in DCE (50 mL). The reaction mixture was stirred at room temperature for 20 min. NaBH(OAc)3 (7.7 g, 36.2 mmol) was added, and the mixture was stirred at room temperature for 16 h. The reaction mixture was quenched by adding MeOH (20 mL) and filtered. The filtrate was concentrated under vacuum, and the residue was purified by reverse HPLC on a C18 / 80 g column (A: water 10 mM NH4HCO3, B: MeOH, 0–100%) to provide 5-(2-(azacyclobutane-1-yl)ethyl)-2-methoxy-4-(trifluoromethyl)pyridine (3 g) as a yellow oil. Yield 63% (ESI 261.2 (M+H)). + ).

[0181] Step 4: 5-(2-(azacyclobutane-1-yl)ethyl)-4-(trifluoromethyl)pyridin-2(1H)-one

[0182]

[0183] A mixture of 5-(2-(azacyclobutan-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 HPLC on a C18 / 80 g column (A: 10 mM water NH4HCO3, B: MeOH, 0–100%) to provide 710 mg of 5-(2-(azacyclobutan-1-yl)ethyl)-4-(trifluoromethyl)pyridine-2(1H)-one as a yellow oil. Yield 25% (ESI 247.1 (M+H)). + ).

[0184] Step 5: Ethyl 2-(5-(2-(azacyclobutane-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoate

[0185]

[0186] Ethyl 4-methyl-2-(methanesulfonyloxy)valerate (1.1 g, 4.4 mmol) and K₂CO₃ (1.2 g, 8.7 mmol) were added to a solution of 5-(2-(azacyclobutan-1-yl)ethyl)-4-(trifluoromethyl)pyridin-1(2H)-one (710 mg, 2.9 mmol) in MeCN (10 mL). The reaction mixture was stirred at 80 °C for 16 h. The reaction mixture was concentrated under vacuum, and the residue was purified by reverse HPLC on a C18 / 40 g column (A: 10 mM water NH₄HCO₃, B: MeOH, 0–100%) to provide ethyl 2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylvalerate (500 mg) as a yellow oil. Yield 44% (ESI 389.2(M+H)) + ).

[0187] Step 6: 2-(5-(2-(azacyclobutane-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoic acid

[0188]

[0189] Ethyl 2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoate (500 mg, 1.3 mmol) was treated with EtOH (5 mL) containing LiOH-H2O (270 mg, 6.5 mmol) 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 HPLC on a C18 / 40 g column (A: 10 mM water NH4HCO3, B: MeOH, 0–100%) to provide 2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoic acid (410 mg) as a yellow oil. Yield 88% (ESI 361.2(M+H)) + ).

[0190] Preparation of 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoic acid

[0191] Step 1: 2-(6-methoxy-4-(trifluoromethyl)pyridin-3-yl)-N,N-dimethylethane-1-amine

[0192]

[0193] Dimethylamine (3.9 mL, 7.8 mmol) and acetic acid (0.05 mL, 0.78 mmol) were added to a solution of 2-(6-methoxy-4-(trifluoromethyl)pyridin-3-yl)acetaldehyde (0.34 g, 1.6 mmol) in DCE (7.8 mL), and the mixture was stirred for 1 hour. Sodium triacetoxyborohydride (0.6 g, 3.1 mmol) was added to the solution. The reaction was stirred for 12 hours, then concentrated and purified by silica gel chromatography (0-35% DCM (1% TEA): MeOH 0-30%) to produce 2-(6-methoxy-4-(trifluoromethyl)pyridin-3-yl)-N,N-dimethylethane-1-amine (305 mg). Yield 79% (ESI 249.27 (M+H)). + ).

[0194] Step 2: 5-(2-(dimethylamino)ethyl)-4-(trifluoromethyl)pyridin-2(1H)-one

[0195]

[0196] HBr (33% in acetic acid) (4.04 mL, 24.57 mmol) was added to 2-(6-methoxy-4-(trifluoromethyl)pyridin-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 provide 5-(2-(dimethylamino)ethyl)-4-(trifluoromethyl)pyridin-2(1H)-one (219 mg). Yield 76% (ESI 235.15(M+H)). + ). 1 H NMR(400MHz,MeOD)δ7.56(s,1H),6.85(s,1H),2.76(m,2H),2.61(m,1H),2.37(m,6H)

[0197] Step 3: Ethyl 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoate

[0198]

[0199] A mixture of 5-(2-(dimethylamino)ethyl)-4-(trifluoromethyl)pyridin-2(1H)-one (685 mg, 2.92 mmol), K₂CO₃ (1.60 g, 11.55 mmol), and ethyl 4-methyl-2-(methanesulfonyloxy)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 chromatography (DCM:MeOH 2:1) to give ethyl 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoate (390 mg) as a brown oil. Yield 35% (ESI 377.2 (M+H)). + ). 1 HNMR(500MHz,MeOD)δ7.84(s,1H),6.68(s,1H),5.51(dd,J=11.0,5.0Hz,1H),4.23(q,J=7.0Hz,2H),2.77(t,J=8.0Hz,2H),2.53(t ,J=8.0Hz,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=7.0Hz,3H),0.97(t,J=7.0Hz,6H).

[0200] Step 4: 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoic acid

[0201]

[0202] Ethyl 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoate (390 mg, 1.0 mmol) was treated with EtOH (10 mL) containing LiOH monohydrate (435 mg, 10.36 mmol) 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 chromatography (MeOH:EtOAc 1:2) to give 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoic acid (358 mg) in oil. Yield 99% (ESI 349.1(M+H)). + ).

[0203] Preparation of 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoic acid

[0204] Step 1: Ethyl 2-(5-(3-(dimethylamino)prop-1-yn-1-yl)2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoate

[0205]

[0206] Ethyl 2-(5-bromo-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoate (68.0 g, 177 mmol, 1.00 equivalent) in THF (408 mL) was supplemented with N,N-dimethylprop-2-yn-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). The reaction mixture was stirred at 25 °C for 12 hours. The reaction mixture was diluted with ethyl acetate, washed twice with saturated ammonium chloride solution, and once with brine. The combined organic phases were dried over 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)prop-1-yn-1-yl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoate (20.0 g, 51.8 mmol, 29.2% yield), which was a yellow oil.

[0207] Step 2: Ethyl 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoate

[0208]

[0209] Pd / C (6.00 g, 2.59 mmol, 5% purity, 0.05 equivalent) was added to a solution of ethyl 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoate (20.0 g, 51.8 mmol, 1 equivalent) in EtOH (200 mL). The mixture was stirred at 15 °C for 24 h under H2 (50 psi). The reaction mixture was filtered and the filtrate was concentrated to give ethyl 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoate (19.7 g, 44.4 mmol, 85.7% yield) as a yellow oil.

[0210] Step 3: 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoic acid

[0211]

[0212] LiOH·H₂O (4.23 g, 101 mmol, 2.00 equivalents) was added to a solution of ethyl 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoate (19.7 g, 50.4 mmol, 1 equivalent) in THF (98.0 mL) and H₂O (20.0 mL) 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 1 N HCl aqueous solution and concentrated under vacuum to give the crude product. The crude product was purified by preparative HPLC (column: Phenomenex luna C18 (250*70mm, 10um); mobile phase: [water (0.05% HCl)-ACN]; B%: 15%-45%, 20 min) to provide 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoic acid (5.40 g, 13.5 mmol, 26.8% yield, HCl) as a white solid. 1 H NMR:400MHz D2Oδ:7.71(s,1H),7.00(s,1H),5.40-5.45(d,J=20Hz,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

[0213] Example 3: Synthesis of exemplary compounds of the present invention

[0214] Preparative HPLC methods

[0215] The crude sample was dissolved in MeOH and purified by preparative HPLC using a Gilson 215 instrument at a detection wavelength of 214 nm.

[0216] Preparative HPLC A: Column: Ultimate C18, 21.2*250mm, 10μm; Mobile phase: Water (10mM ammonium bicarbonate), BCH3CN; Gradient elution as described in the text; Flow rate: 30 mL / min.

[0217] 3-52. Preparation of (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propionic acid (compounds HH-P1 and HH-P2)

[0218] Step 1: Ethyl (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propionate

[0219]

[0220] Ethyl (3S)-3-amino-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propionate (120 mg, 0.3 mmol), 2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-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) were stirred overnight at room temperature. The reaction was concentrated under vacuum, and the residue was analyzed by reverse HPLC on a C18 / 40 g column (A: 10 mM water NH4HCO3). 3，Purification was performed on (B: MeOH, 20–95%) to provide ethyl (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propionate (140 mg) as a pale yellow solid. Yield 62% (ESI 734.2 [M+H) + ).

[0221] Step 2: (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propionic acid

[0222]

[0223] Ethyl (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propionate (140 mg, 0.19 mmol) was treated with EtOH (2 mL) containing LiOH-H2O (24 mg, 0.57 mmol) 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 was concentrated under vacuum, and the residue was purified by preparative HPLC A (20–85% MeCN) to give diastereomers HH-P1 (35 mg) and HH-P2 (58 mg) as white solids.

[0224] HH-P1 ESI 706.2(M+H) +1 H NMR(400MHz,MeOD)δ7.80(s,1H),7.12–7.03(m,1H),7.00–6.91(m,1H),6.87 (s,1H),6.62(t,J=8.0Hz,1H),5.80–5.58(m,2H),4.00(t,J=7.7Hz,4H),3.3 0–3.19(m,2H),2.97–2.78(m,3H),2.71–2.62(m,1H),2.49–2.34(m,2H),2.1 3–1.76(m,9H),1.46–1.33(m,1H),1.00–0.86(m,8H),0.66(d,J=4.8Hz,2H).

[0225] HH-P2 ESI 706.2(M+H) +1 H NMR(400MHz,MeOD)δ7.70(s,1H),7.14–7.04(m,1H),7.02–6.87(m,2H),6.67(t,J=8.1H z,1H),5.98–5.86(m,1H),5.62(t,J=7.7Hz,1H),4.11(t,J=8.1Hz,4H),3.45–3.33(m,2 H),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).

[0226] 3-57. Preparation of (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propionic acid (compounds HM-P1 and HM-P2)

[0227] Step 1: Ethyl (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propionate

[0228]

[0229] The mixture of (S)-3-amino-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propionate (120 mg, 0.31 mmol), 2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-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 chromatography (DCM:MeOH 97:3) to provide ethyl (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propionate (120 mg) as a colorless oil. Yield 53% (ESI 730.3(M+H) + ).

[0230] Step 2: (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propionic acid

[0231]

[0232] Ethyl (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethylbiphenyl-3-yl)propionate (120 mg, 0.16 mmol) was treated with EtOH (3 mL) and H2O (1 mL) containing LiOH-H2O (35 mg, 0.9 mmol) at room temperature for 2 hours. The reaction mixture was acidified to pH 4–5 with 1N HCl. The reaction was concentrated under vacuum, and the residue was purified by preparative HPLC A (30–60% CH3CN) to give diastereomers HM-P1 (30.2 mg) and HM-P2 (31.8 mg) as white solids.

[0233] HM-P1 ESI 702.3(M+H) +. 1H NMR(400MHz,MeOD)δ7.80(s,1H),6.97–6.79(m,3H),6.58(t,J=8.1Hz,1H),5.84–5.61(m,2H),4.00(t,J=8.1Hz,4H),3.30–3.21(m,2H),2.88-2.85 (m,3H),2.67(dd,J=14.8,4.8Hz,1H),2.48–2.37(m,2H),2.28(d,J=6.4H z,3H),2.13–1.96(m,3H),1.90(d,J=21.3Hz,6H),1.46–1.31(m,1H),0.94 -0.90(m,8H),0.64-0.62(m,2H).

[0234] HM-P2 ESI 702.3(M+H) + . 1H NMR(400MHz,MeOD)δ7.70(s,1H),7.00–6.86(m,3H),6.63(t,J=8.2Hz,1H),5.93 (dd,J=11.4,3.5Hz,1H),5.62(t,J=7.7Hz,1H),4.11(t,J=8.0Hz,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=11.4,6.6Hz,6H),0.67-0.65(m,2H).

[0235] 3-62. Preparation of (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid (compounds HR-P1 and HR-P2)

[0236] Step 1: (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)ethyl propionate

[0237]

[0238] Ethyl (3S)-3-amino-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (90 mg, 0.21 mmol), 2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-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) were stirred at room temperature for 2 hours. The solvent was concentrated under vacuum, and the residue was purified by reversed-phase HPLC on a C18 / 40g column (A: 10mM water NH4HCO3, B: CH3CN, 0–100%) to provide ethyl (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (80 mg) as a white solid. Yield 50% (ESI 776.2 [M+H) + ).

[0239] Step 2: (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid

[0240]

[0241] Ethyl (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (80 mg, 0.1 mmol) was treated with MeOH (2.0 mL) and H2O (0.5 mL) containing LiOH-H2O (13 mg, 0.3 mmol) at room temperature for 2 hours. The reaction mixture was acidified to pH 5–6 with 1 N HCl. The reaction was concentrated in a vacuum, and the residue was purified by preparative HPLC A (50-80% CH3CN) to give diastereomer products HR-P1 (22.0 mg) and HR-P2 (25.0 mg) as white solids.

[0242] HR-P1 ESI 748.2 (M+H) + . 1 H NMR (400MHz, MeOD) δ7.79(s,1H),7.40(t,J=7.5Hz,1H),6.99(d,J=7.9Hz,1H),6.85(d,J=3.0Hz,1H),5.78–5.60(m,2H),4.04(t,J=8.0Hz,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=1.6Hz,3H),2.09–1.75(m,8H),1.37(s,1H),0.93(t,J=6.4Hz,6H).

[0243] HR-P2 ESI 748.2(M+H) + . 1 H NMR (400MHz, MeOD) δ7.72(s,1H),7.46(t,J=7.5Hz,1H),7.03(d,J=7.6Hz,1H),6.91( s,1H),5.87–5.82(m,1H),5.63(t,J=7.7Hz,1H),4.12(t,J=7.8Hz,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=1.5Hz,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).

[0244] 3-65. Preparation of (3S)-3-(4'-cyclopropyl-2,4-difluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-(3-(5-(2-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)propionic acid (compounds HU-P1 and HU-P2)

[0245] 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)pyridin-1(2H)-yl)-4-methylpentamido)propionate

[0246]

[0247] Ethyl (S)-3-amino-3-(4'-cyclopropyl-2,4-difluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (140.0 mg, 0.32 mmol), 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-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) were stirred overnight at room temperature. The reaction was concentrated under vacuum, and the residue was purified by reverse HPLC on a C18 / 40g column (A: 10mM water NH4HCO3, B: MeOH, 0–100%) to provide 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)pyridin-1(2H)-yl)-4-methylpentamido)propionate (150.0 mg) as a pale yellow solid. Yield 60% (ESI 786.3 [M+H)). + ).

[0248] 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)pyridin-1(2H)-yl)-4-methylpentamido)propionic acid

[0249]

[0250] 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)pyridin-1(2H)-yl)-4-methylpentamido)propionate (150.0 mg, 0.19 mmol) was treated with EtOH (2 mL) containing LiOH-H2O (23.9 mg, 0.57 mmol) 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 was concentrated under vacuum, and the residue was purified by preparative HPLC A (30–80% MeCN) to give diastereomers HU-P1 (54.5 mg) and HU-P2 (50.5 mg) as white solids.

[0251] HU-P1 ESI 758.3(M+H) +1 H NMR(400MHz,MeOD)δ7.76(s,1H),7.34(t,J=7.6Hz,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).

[0252] HU-P2 ESI 758.3(M+H) +1 H NMR(400MHz,MeOD)δ7.79(s,1H),7.39(t,J=7.6Hz,1H),6.85(s,3H),5.85 –5.79(m,1H),5.62(t,J=7.6Hz,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).

[0253] 3-67. Preparation of (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid (compounds HW-P1 and HW-P2)

[0254] Step 1: Ethyl (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate

[0255]

[0256] A mixture of 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-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)propionate (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 was concentrated under vacuum, and the residue was purified by silica gel column chromatography (DCM:MeOH 4:1) to provide ethyl (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (150.0 mg) as a brown solid. Yield 71% (ESI 764.7(M+H)) + ).

[0257] Step 2: (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid

[0258]

[0259] Ethyl (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (150 mg, 0.20 mmol) was treated with MeOH (4 mL) and H2O (1 mL) containing LiOH-H2O (33.6 mg, 0.80 mmol) at room temperature for 2 hours. The reaction mixture was acidified to pH 4–5 with 1N HCl. The reaction was concentrated under vacuum, and the residue was purified by preparative HPLC A (30–60% MeCN) to give diastereomers HW-P1 (60.1 mg) and HW-P2 (50.0 mg) as white solids.

[0260] HW-P1 ESI 736.6(M+H) + . 1H NMR(400MHz,MeOD)δ7.76(s,1H),7.39(t,J=7.5Hz,1H),6.88(d,J=9.3Hz,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=6.6Hz,6H).

[0261] HW-P2 ESI 736.6(M+H) + . 1H NMR(400MHz,MeOD)δ7.79(s,1H),7.45(t,J=7.6Hz,1H),6.96–6.80(m,3H),5.86–5.75(m,1H),5.62(t,J=7.6Hz,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).

[0262] 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)pyridin-1(2H)-yl)-4-methylpentamido)propionic acid (compounds HZ-P1 and HZ-P2)

[0263] Step 1: Ethyl (3S)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)propionate

[0264]

[0265] Ethyl (S)-3-amino-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propionate (208 mg, 0.50 mmol), 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylvaleric acid (180 mg, 0.50 mmol), TCFH (280 mg, 1.0 mmol), and NMI (123 mg, 1.5 mmol) in MeCN (5 mL) were stirred at room temperature for 3 hours. The solvent was removed under vacuum, and the residue was purified by silica gel column chromatography (DCM:MeOH 97:3) to provide ethyl (3S)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)propionate (200 mg) as a colorless oil. Yield 53% (ESI 760.3(M+H) + ).

[0266] 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)pyridin-1(2H)-yl)-4-methylpentamido)propionic acid

[0267]

[0268] Ethyl (3S)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)propionate (200 mg, 0.26 mmol) was treated with EtOH (3 mL) and H2O (1 mL) containing LiOH-H2O (55 mg, 1.3 mmol) at room temperature for 2 hours. The reaction mixture was acidified to pH 4–5 with 1N HCl. The reaction was concentrated under vacuum, and the residue was purified by preparative HPLC A (30–60% CH3CN) to give diastereomers HZ-P1 (7.0 mg) and HZ-P2 (62.0 mg) as white solids.

[0269] HZ-P1 ESI 732.3(M+H) + . 1H NMR(400MHz,MeOD)δ7.76(s,1H),7.35(t,J=7.6Hz,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=6.5Hz,6H).

[0270] HZ-P2 ESI 732.3(M+H) + . 1H NMR(400MHz,MeOD)δ7.79(s,1H),7.40(t,J=7.6Hz,1H),6.97(s,2H),6.85(s,1H),5.82-5 .80(m,1H),5.62(t,J=7.6Hz,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).

[0271] 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)pyridin-1(2H)-yl)-4-methylpentamido)propionic acid (compounds IF-P1 and IF-P2).

[0272] Step 1: (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)pyridin-1(2H)-yl)-4-methylpentamido)propionate

[0273]

[0274] The mixture of (S)-3-amino-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (210 mg, 0.5 mmol), 2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-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 was concentrated and purified by reversed-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)pyridin-1(2H)-yl)-4-methylpentamido)propionate (330 mg) as a yellow solid. Yield 88.5% (ESI 746.3 [M+H)). + ).

[0275] 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)pyridin-1(2H)-yl)-4-methylpentamido)propionic acid

[0276]

[0277] 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)pyridin-1(2H)-yl)-4-methylpentamido)propionate (330 mg, 0.44 mmol) was treated with MeOH (4 mL) and H2O (1 mL) containing LiOH-H2O (56 mg, 1.34 mmol) at room temperature for 30 min. The reaction mixture was acidified to pH 5-6 with 1N HCl. The reaction was concentrated under vacuum, and the residue was purified by preparative HPLC A (30-58% CH3CN) to give diastereomers IF-P1 (96 mg) and IF-P2 (94 mg) as white solids.

[0278] IF-P1 ESI 718.3(M+H) + . 1 H NMR(400MHz,MeOD)δ7.86(s,1H),7.34(t,J=7.6Hz,1H),6.94(d,J=3.7Hz,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=7.0Hz,6H).

[0279] IF-P2 ESI 718.3(M+H) + . 1H NMR(400MHz,MeOD)δ7.84(s,1H),7.41(t,J=7.6Hz,1H),6.97(s,2H),6.89(s,1H), 5.85–5.76(m,1H),5.66(t,J=7.8Hz,1H),3.25–3.09(m,2H),2.96(t,J=7.2Hz,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=3.1Hz ,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).

[0280] 3-77. Preparation of (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-biphenyl-3-yl)propionic acid (compounds IG-P1 and IG-P2)

[0281] Step 1: (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propionate ethyl ester

[0282]

[0283] A mixture of 2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentanoic acid (150 mg, 0.41 mmol), ethyl (S)-3-amino-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propionate (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 preparative HPLC A (30-90% CH3CN) to provide ethyl (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)biphenyl-3-yl)propionate (150 mg) as a white solid. Yield 54% (ESI 758.2 [M+H)). + ).

[0284] Step 2: (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-biphenyl-3-yl)propionic acid

[0285]

[0286] Ethyl (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-biphenyl-3-yl)propionate (150 mg, 0.19 mmol) was treated with MeOH (4 mL) and H2O (0.4 mL) containing LiOH-H2O (42 mg, 1.00 mmol) at room temperature for 2 hours. The reaction mixture was acidified to pH 4–5 with 1N HCl. The reaction was concentrated under vacuum, and the residue was purified by preparative HPLC A (30–70% CH3CN) to give diastereomers IG-P1 (33.0 mg) and IG-P2 (51.0 mg) as white solids.

[0287] IG-P1 ESI 730.2(M+H) + . 1 H NMR(400MHz,MeOD)δ7.79(s,1H),7.37(t,J=7.5Hz,1H),6.95(s,2H),6.85(s,1H),5.79–5.65(m,2H),4.03(t,J=8.1Hz,4H),3.2 7–3.20(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).

[0288] IG-P2 ESI 730.2(M+H) + . 1 H NMR(400MHz,MeOD)δ7.73(s,1H),7.43(t,J=7.6Hz,1H),6.97(s,2H),6.91(s, 1H),6.00–5.83(m,1H),5.63(t,J=7.7Hz,1H),4.11(t,J=8.0Hz,4H),3.50–3.3 3(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).

[0289] 3-86. Preparation of (3S)-3-(2-(5-(3-(azacyclobutan-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid (compounds IP-P1 and IP-P2).

[0290] Step 1: Ethyl (3S)-3-(2-(5-(3-(azacyclobutan-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate

[0291]

[0292] Ethyl (S)-3-amino-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (112.0 mg, 0.27 mmol), 2-(5-(3-(azacyclobutan-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridin-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) were stirred at room temperature for 1 hour. The reaction was concentrated under vacuum, and the residue was purified by silica gel column chromatography (DCM:MeOH 4:1) to provide ethyl (3S)-3-(2-(5-(3-(azacyclobutane-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (130.0 mg) as a brown solid. Yield 63% (ESI 772.6(M+H)) + ).

[0293] Step 2: (3S)-3-(2-(5-(3-(azacyclobutan-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid

[0294]

[0295] Ethyl (3S)-3-(2-(5-(3-(azacyclobutan-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (130 mg, 0.17 mmol) was treated with MeOH (4 mL) and H2O (1 mL) containing LiOH-H2O (28.6 mg, 0.68 mmol) at room temperature for 2 hours. The reaction mixture was acidified to pH 4–5 with 1N HCl. The reaction was concentrated under vacuum, and the residue was purified by preparative HPLC A (30–60% MeCN) to give diastereomers IP-P1 (38.0 mg) and IP-P2 (42.0 mg) as white solids.

[0296] IP-P1 ESI 744.7(M+H) + . 1H NMR(400MHz,MeOD)δ7.76(s,1H),7.34(d,J=7.4Hz,1H),6.94(s,1H),6.79(s,2H),5.74(d,J=4.1Hz,2H),4.08(t,J=7.7Hz,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 = 43.4Hz, 5H), 1.34 (s, 1H), 0.93 (d, J = 6.5Hz, 6H).

[0297] IP-P2 ESI 744.7(M+H) + . 1H NMR(400MHz,MeOD)δ7.78(s,1H),7.40(t,J=7.7Hz,1H),6.90(d,J=55.2Hz,3 H),5.87–5.78(m,1H),5.61(t,J=7.6Hz,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).

[0298] 3-93. Preparation of (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propionic acid (compounds IW-P1 and IW-P2)

[0299] Step 1: Ethyl (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propionate

[0300]

[0301] Ethyl (3S)-3-amino-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propionate (170 mg, 0.45 mmol), 2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-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) were stirred overnight at room temperature. The reaction was concentrated under vacuum, and the residue was purified by reverse HPLC on a C18 / 40g column (A: 10mM water NH4HCO3, B: MeOH, 20–95%) to provide ethyl (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propionate (210.0 mg) as a pale yellow solid. Yield 65% (ESI 724.2 [M+H) + ).

[0302] Step 2: (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propionic acid

[0303]

[0304] Ethyl (3S)-3-(2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propionate (210.0 mg, 0.29 mmol) was treated with EtOH (2 mL) containing LiOH-H2O (36.5 mg, 0.87 mmol) 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 was concentrated under vacuum, and the residue was purified by preparative HPLC A (20–85% MeCN) to give diastereomers IW-P1 (68 mg) and IW-P2 (52 mg) as white solids.

[0305] IW-P1 ESI 696.2 (M+H) +1 H NMR(400MHz,MeOD)δ7.79(s,1H),6.98–6.80(m,3H),5.83–5.69(m,2H),3.09(t,J=7.9Hz,2H),3.00–2.90(m,1H),2.81 (d,J=0.8Hz,6H),2.73–2.58(m,3H),2.26(d,J=1.3Hz,6H),2.15–1.78(m,10H),1.43–1.30(m,1H),1.00–0.90(m,6H).

[0306] IW-P2 ESI 696.2(M+H) +1 H NMR(400MHz,MeOD)δ7.79(s,1H),7.02–6.79(m,3H),5.89–5.79(m,1H),5.57(t,J=7.6Hz,1H),3.09–2.95(m,2H),2.91–2.7 1(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).

[0307] 3-142. Preparation of (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid (compounds KT-P1 and KT-P2)

[0308] Step 1: (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)ethyl propionate

[0309]

[0310] Ethyl (3S)-3-amino-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (140 mg, 0.32 mmol), 2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-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) were stirred at room temperature for 2 hours. The solvent was removed under vacuum, and the residue was purified by reversed-phase HPLC on a C18 / 40g column (A: 10mM water NH4HCO3, B: CH3OH, 0–85%) to provide ethyl (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (140 mg) as a yellow solid. Yield 56.4% (ESI 776.3 [M+H)). + ).

[0311] Step 2: (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid

[0312]

[0313] Ethyl (3S)-3-(2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionate (140 mg, 0.18 mmol) was treated with EtOH (4 mL) and H2O (1 mL) containing LiOH-H2O (22 mg, 0.54 mmol) 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 preparative HPLC A (30–65% CH3CN) to provide diastereomers KT-P1 (24 mg) and KT-P2 (30 mg) as white solids.

[0314] KT-P1 ESI 748.2(M+H) + . 1 H NMR(400MHz,MeOD)δ7.81(s,1H),7.39(m,J=7.5Hz,1H),6.99–6.80(m,2H),5.71(m,J=9.2,5.9Hz,2H),4.12(m,J=8.1Hz,4H),3.42–3.3 0(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=6.9Hz,6H).

[0315] KT-P2 ESI 748.2(M+H) + . 1 H NMR(400MHz,MeOD)δ7.72(s,1H),7.44(m,J=7.6Hz,1H),7.00–6.77(m,2H),5.90(d,J=8.1Hz,1H) ,5.63(m,J=7.7Hz,1H),4.12(m,J=8.1Hz,4H),3.39(m,J=19.1,13.7Hz,2H),2.86(m,J=14.4,13. 3Hz,3H),2.60(m,J=15.6,4.2Hz,1H),2.45(m,J=8.1Hz,2H),2.20(s,3H),1.94(m,J=21.4,9.3,4 .8Hz, 7H), 1.72 (m, J = 14.4, 7.3Hz, 1H), 1.33 (m, J = 13.4, 6.7Hz, 1H), 0.88 (m, J = 10.6, 6.6Hz, 6H).

[0316] Brief introduction to the in vitro assays described in Examples 4-6

[0317] Three in vitro assays were used to examine the mechanistic processes by which cells utilize α4β7: 1) ligand:receptor affinity; 2) the affinity of these interactions at the cell surface; and 3) how these interactions behave under applied forces. In Example 4, a fluorescence polarization (FP) assay was used to measure compound activity by competing with the binding of a fluorescein-labeled peptide. In Example 5, the potency of a compound for α4β7 was measured in a cell-based ligand binding assay (LBA) using RPMI 8866 cells incubated with a compound sample competing with a soluble MAdCAM-1 ligand. In Example 6, the activity of the compound was evaluated in a cell adhesion assay, which mechanically tests what happens in vivo when transport cells utilize α4β7 to adhere to MAdCAM-1-expressing HEVs in the gut during extravasation. In the cell adhesion assay of Example 6, MAdCAM1-(Fc) was coated onto a plastic, and α4β7-expressing cells (RPMI-8866) were allowed to adhere to the coated surface in the presence of the test compound. Next, a force of washing with buffer is applied to the cells to test the strength of the adhesion. Unattached cells are removed, and the remaining adherent cells are quantified.

[0318] Example 4: Fluorescence polarization determination of the α4β7 binding of a compound

[0319] Fluorescence polarization (FP) assays were used to measure compound activity by competition with the binding of the fluorescein-labeled peptide CRSDTLCGE{Lys(FITC)}. In the assay, 6.5 nM integrin α4β7 was incubated with the test compound in a solution containing 2 mM manganese chloride, 0.1 mM calcium chloride, 20 mM HEPES buffer (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 384-well plates. Before adding the fluorescein-labeled peptide, the integrin and test compound were pre-incubated at 22°C for 15 min. After adding the fluorescein-labeled peptide, the assay was incubated at 22°C for 1 h, and the fluorescence polarization was measured. The IC50 was determined by nonlinear regression and four-parameter curve fitting. 50 value.

[0320] Using the FP measurement in Example 4 to include Figure 1 The inhibitory efficacy of certain compounds against α4β7 was measured. Figure 1 The IC obtained using the FP measurement in Example 4 is provided. 50The values ​​are given as a range (A: <5nM; B: 5nM to 500nM; C: >500nM). Figure 1 middle).

[0321] The α4β7 inhibitory efficacy of the compounds in Tables 1 and 2 (comparative) were also measured using the FP assay of Example 4, and the results are provided as IC50 values. 50 The numerical range of values ​​(A: ≤10nM; B: >10nM to 500nM; C: >500nM, in Tables 1 and 2).

[0322] Example 5: Ligand Binding Assay

[0323] To measure the potency of the compound for α4β7 in a cell-based ligand binding assay (LBA), RPMI 8866 cells were incubated with 10 μl of the compound sample at room temperature for 15 min in a buffer containing 50 mM HEPES pH 7.3, 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 immobilizable active dye. 5 μl of 33 nM MAdCAM-1-Fc, fluorescently labeled with Dylight 650, was added to the cells in a solution of 50 mM HEPES pH 7.3, 150 mM sodium chloride, and 1% bovine serum albumin. The sample was incubated at room temperature for 45 min, fixed with 0.8% formaldehyde at room temperature for 30 min, and washed with 50 mM Tris pH 7.5, 150 mM NaCl, 1 mM EDTA, and 1% bovine serum albumin. Fluorescence intensity of each cell was measured by flow cytometry. Dead cells were excluded from further analysis based on staining with a 780-type fixable reactive dye. The median fluorescence intensity of Dylight 650 for each sample was determined, and concentration-response curves with IC50 values ​​were analyzed using a 4-parameter nonlinear regression analysis.

[0324] The ligand binding assay was performed using Example 5. Figure 1 The α4β7 ligand binding assays of the compounds listed were performed. Figure 1 In the example, IC was obtained using the LB measurement of Example 5. 50 The values ​​are provided by a numerical range (A: <5nM; B: 5nM to 500nM; C: >500nM). Figure 1 middle).

[0325] The α4β7 ligand binding assays of the compounds in Table 1 and (Comparative) Table 2 were also performed using the ligand binding assays of Example 5, and the resulting IC50 values ​​were... 50 Values ​​are provided as numerical ranges (A: ≤10nM; B: >10nM to 500nM; C: >500nM, in Tables 1 and 2).

[0326] Example 6: Cell Adhesion Assay

[0327] Example 6 describes a cell adhesion assay. α4β7 cell adhesion measurements from the assays in Example 6 were obtained from the compounds in Table 1 and the comparative compounds in Table 2, and the results are presented as the obtained IC50 values. 50 The numerical range of values ​​(A: <5nM; B: 5nM to <10nM; C: 10-50nM; D: >50nM; E: >100nM and F: >500nM, in Tables 1 and 2).

[0328] Add 100 μL of PBS containing 100 μg of recombinant human MAdCAM to each well of a 96-well plate and incubate overnight at 4°C. After incubation, remove MAdCAM by aspiration and add 200 μL of PBS + 1% BSA to block the plate at 37°C and 5% CO2 for 2 hours. During this incubation, prepare a dilution profile of the compound in 100% DMSO in a 96-well V plate. Then transfer 1.75 μL of the diluted compound to a new 96-well U plate containing 20 μL of assay medium (phosphate-free DMEM + 25 mM HEPES + 1% BSA). Add an additional 155 μL of assay medium to this plate by pipetting. Allow this mixture to incubate at 37°C and 5% CO2 for 15 minutes. After incubation, add 175 μL of assay medium containing 2e6 / mL RPMI8866 cells to the well containing the compound without mixing, and allow the plate to incubate an additional 15 minutes at 37°C and 5% CO2. During this incubation, the MAdCAM-coated plate was removed from the incubator and washed twice with 200 μL of PBS + 0.1% BSA. After incubating the cells with the compound for 15 minutes, the mixture was mixed by pipetting, and 100 μL of the mixture was transferred in triplicate to the washed MAdCAM-coated plate. This plate was then incubated at 37°C and 5% CO2 for 1 hour. After incubation, the plate was washed twice with 200 μL and once with 50 μL of phenol-free RPMI + 1% BSA. After the final wash, the final 50 μL of phenol-free RPMI + 1% BSA was added to the wells. Next, 50 μL of Promega cell titer luminescence was added to the wells. The plate was incubated on a shaker at 200 RPM for 2 minutes, then incubated on a shaker for an additional 8 minutes, after which the luminescence was read on a Biotek Citation 5 microplate reader. The raw data were converted to % inhibition compared to the bottom of the curve and analyzed using a 4-parameter nonlinear curve in Prism to determine the IC50. 50 and IC 90 .

[0329] Table 1. Selected exemplary compounds.

[0330]

[0331]

[0332]

[0333] Table 2. Comparative Compounds (See PCT International Application Publication No. WO 2019 / 200202; the international application publication is incorporated by reference)

[0334]

[0335]

[0336]

[0337]

[0338]

[0339]

[0340]

[0341] By incorporating via reference

[0342] All U.S. patents and U.S. and PCT patent application publications cited herein are incorporated herein by reference.

[0343] Equivalent solution

[0344] Those skilled in the art will recognize that many equivalent forms of the specific embodiments of the invention described herein can be determined using only conventional experiments. Such equivalent forms are intended to be covered by the following claims.

Claims

1. A pharmaceutical composition comprising a compound selected from the group consisting of: ； ； ； ； ； ； ； ； ； ;as well as 。 2. The pharmaceutical composition according to claim 1, wherein the compound is Or its pharmaceutically acceptable salt.

3. The pharmaceutical composition according to claim 1, wherein the compound is Or its pharmaceutically acceptable salt.

4. The pharmaceutical composition according to claim 1, wherein the compound is Or its pharmaceutically acceptable salt.

5. The pharmaceutical composition according to claim 1, wherein the compound is Or its pharmaceutically acceptable salt.

6. The pharmaceutical composition according to claim 1, wherein the compound is Or its pharmaceutically acceptable salt.

7. The pharmaceutical composition according to claim 1, wherein the compound is Or its pharmaceutically acceptable salt.

8. The pharmaceutical composition according to claim 1, wherein the compound is Or its pharmaceutically acceptable salt.

9. The pharmaceutical composition according to claim 1, wherein the compound is Or its pharmaceutically acceptable salt.

10. The pharmaceutical composition according to claim 1, wherein the compound is Or its pharmaceutically acceptable salt.

11. The pharmaceutical composition according to claim 1, wherein the compound is Or its pharmaceutically acceptable salt.

12. The pharmaceutical composition according to claim 1, wherein the compound is Or its pharmaceutically acceptable salt.

13. A compound selected from the group consisting of: (S)-3-((S)-2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethyl-[1,1'-biphenyl]-3-yl)propionic acid; (3S)-3-((S)-2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propionic acid; (S)-3-((S)-2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid; (3S)-3-((S)-2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propionic acid; (3S)-3-((S)-2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid; (S)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-((S)-2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)propionic acid; (S)-3-((S)-2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid; (S)-3-((S)-2-(5-(3-(azacyclobutan-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid; (S)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-((S)-2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)propionic acid; (S)-3-(4'-cyclopropyl-2,4-difluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-((S)-2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)propionic acid; and (3S)-3-((S)-2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid, Or its pharmaceutically acceptable salt.

14. The compound according to claim 13, wherein the compound is (S)-3-((S)-2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,4-difluoro-2',4',6'-trimethyl-[1,1'-biphenyl]-3-yl)propionic acid, Or its pharmaceutically acceptable salt.

15. The compound according to claim 13, wherein the compound is (3S)-3-((S)-2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(5-cyclopropyl-2,3',4-trifluoro-2',6'-dimethyl-[1,1'-biphenyl]-3-yl)propionic acid, Or its pharmaceutically acceptable salt.

16. The compound according to claim 13, wherein the compound is (S)-3-((S)-2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid, Or its pharmaceutically acceptable salt.

17. The compound according to claim 13, wherein the compound is (3S)-3-((S)-2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',5,6'-tetramethyl-[1,1'-biphenyl]-3-yl)propionic acid, Or its pharmaceutically acceptable salt.

18. The compound according to claim 13, wherein the compound is (3S)-3-((S)-2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4,4'-trifluoro-2',3',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid, Or its pharmaceutically acceptable salt.

19. The compound according to claim 13, wherein the compound is (S)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-((S)-2-(5-(2-(dimethylamino)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)propionic acid, Or its pharmaceutically acceptable salt.

20. The compound according to claim 13, wherein the compound is (S)-3-((S)-2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid, Or its pharmaceutically acceptable salt.

21. The compound according to claim 13, wherein the compound is (S)-3-((S)-2-(5-(3-(azacyclobutan-1-yl)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid, Or its pharmaceutically acceptable salt.

22. The compound according to claim 13, wherein the compound is (S)-3-(2,4-difluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-((S)-2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)propionic acid, Or its pharmaceutically acceptable salt.

23. The compound according to claim 13, wherein the compound is (S)-3-(4'-cyclopropyl-2,4-difluoro-2',6'-dimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)-3-((S)-2-(5-(3-(dimethylamino)propyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)propionic acid, Or its pharmaceutically acceptable salt.

24. The compound according to claim 13, wherein the compound is (3S)-3-((S)-2-(5-(2-(azacyclobutan-1-yl)ethyl)-2-oxo-4-(trifluoromethyl)pyridin-1(2H)-yl)-4-methylpentamido)-3-(2,3',4-trifluoro-2',4',6'-trimethyl-5-(trifluoromethyl)-[1,1'-biphenyl]-3-yl)propionic acid, Or its pharmaceutically acceptable salt.

25. A pharmaceutical composition comprising, as an active pharmaceutical ingredient, a compound or a pharmaceutically acceptable salt thereof according to any one of claims 13 to 24; and a pharmaceutically acceptable excipient.

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