Compounds and their use as MIF inhibitors

Abstract

The present invention provides a compound of formula I that can be used as an inhibitor of macrophage migration inhibitory factor (MIF); a method for preparing the compound of the present invention; a pharmaceutical composition comprising the compound of the present invention; and the use and method of treating MIF-mediated diseases by administering the compound of the present invention.

Description

Background of the Invention Technical Field

[0002] This invention relates to novel compounds; methods for preparing the compounds of the invention; pharmaceutical compositions comprising the compounds of the invention; and uses and methods for treating diseases mediated by macrophage migration inhibitory factor (MIF) by administering the compounds of the invention. Specifically, the compounds of the invention can be used as MIF inhibitors. Background Technology

[0004] Macrophage migration inhibitory factor (MIF) was initially identified as a cytokine released from active T cells to inhibit the random movement of macrophages. It is secreted by epithelial cells, endothelial cells, lymphocytes, monocytes, and macrophages, suggesting a role in both innate and adaptive immunity. In humans, the MIF gene is located on chromosome 22q11.2 and encodes an evolutionarily conserved protein of 115 amino acids. The monomeric form of the MIF protein has a molecular weight of 12.5 kDa. When activated, MIF forms a trimer composed of three identical subunits, each monomer containing two antiparallel α-helices stacked on a four-stranded β-sheet. MIF exhibits chemokine-like function and has been identified as a ligand for CD74, forming complexes with CXCR2 and CXCR4. Binding of MIF to these receptors enhances monocyte recruitment and leukocyte chemotaxis. The inflammatory cascade depends on the activation of CXCR2 and CD74, indicating that MIF functions through a functional CXCR2 / CD74 complex.

[0005] MIFs have multiple biological functions, most importantly related to inflammation and immunity. MIFs counterregulate the effects of glucocorticoids, naturally occurring steroid hormones produced by the adrenal glands during cellular stress and possessing anti-inflammatory properties. MIFs may stimulate the expression of other cytokines associated with inflammation. Inflammation is essential for survival, but when misregulated, it can lead to tumorigenesis. MIFs play a role in both innate and adaptive immunity and are constitutively expressed by monocytes, macrophages, dendritic cells, B cells, neutrophils, eosinophils, mast cells, and basophils. They promote T cell response to stimuli from exogenous factors and act as a regulator of infection responses by increasing TLR4 expression. Activated T cells release MIFs to suppress glucocorticoid-mediated production of interleukin-2 and interferon-γ. Because circulating glucocorticoid levels increase during infection and inflammation, MIFs exert their immunosuppressive effects, thereby inducing a primary immune response and reducing the need for steroid therapy. MIFs have also been reported to possess enzymatic activity, converting D-dopachrome into 5,6-dihydroxy-2-indolecarboxylic acid (DHICA). Although identifying DHICA as a true biological substrate for MIFs elucidates this mechanism of action, the role of MIF enzymatic activity remains not fully understood.

[0006] MIF is a pluripotent and pleiotropic cytokine expressed in various human malignancies such as glioblastoma, lung cancer, breast cancer, gastric cancer, bladder cancer, and melanoma. In numerous studies, MIF has been shown to contribute to many different forms of cancer. First, MIF is a regulator of the p53 signaling pathway and can physically interact with p53. MIF inhibits p53 activity, leading to dysregulation of the normal cell cycle. Because MIF functionally inactivates p53, cell cycle arrest and apoptosis do not occur. Second, MIF participates in the phosphoinositol-3-kinase (PI3K) / Akt pathway, which plays a crucial role in tumor development. Activation of this pathway prevents apoptosis in key cells. Previous studies have shown that MIF and CD74 initiate Akt activation, and when MIF is overexpressed, it causes key cells to advance through the cell cycle via the PI3K / Akt pathway. Third, MIF plays a role in angiogenesis. When MIF is highly expressed, vascular endothelial growth factor (VEGF), hypoxia-inducible factor 1 (HIF-1), and other angiogenic factors are responsible for new blood vessel formation. Fourth, MIF leads to tumor cell metastasis by decreasing E-cadherin expression and increasing N-cadherin expression. Decreased E-cadherin expression also promotes epithelial-mesenchymal transition (EMT) and may lead to the formation of secondary tumors. EMT is a process that regulates the acquisition of mesenchymal cell characteristics by epithelial cells, thereby leading to invasion and metastasis.

[0007] Beyond the field of oncology, MIF is a pleiotropic inflammatory cytokine with upstream regulatory roles in both innate and adaptive immunity, and is involved in the pathogenesis of autoimmune diseases including multiple sclerosis (MS), rheumatoid arthritis (RA), and systemic lupus erythematosus (SLE). MIF is significantly upregulated in the serum of MS, RA, and SLE patients and animal models. MIF inhibition has been shown to improve the in vitro and in vivo characteristics of autoimmune diseases.

[0008] The significant role of MIFs in cancer development and autoimmune diseases suggests that targeted inhibition of MIFs is a potential therapeutic approach for cancer, autoimmune diseases, and other inflammatory conditions. One of the most important strategies for targeting MIFs is the development of small molecule inhibitors, which have demonstrated promising preclinical efficacy. Summary of the Invention

[0009] This invention provides novel compounds that can be used as MIF inhibitors.

[0010] In one aspect, the present invention provides compounds of formula I:

[0011]

[0012] in

[0013] A1 is a 5- or 6-membered carbon ring or heterocycle.

[0014] A1 is optionally selected by 0, 1, 2 or 3 independently from OH, halogen or C. 1-6 Alkyl substituents;

[0015] A2 is selected from

[0016] n1 is 0 or 1.

[0017] When n1 is 0, R 1 and R 2 It does not exist when n1 is 1, R 1 and R 2 Each of them is independently selected from H, C 1-6 Alkyl, or R 1 and R 2 Together with the carbon atoms it is bonded to, they form C 3-6 cycloalkyl;

[0018] m1 is 0 or 1, m2 is 0, 1 or 2.

[0019] When m1 is 0 and m2 is 1, Y is C; when m1 is 0 and m2 is 2, Y is S; when m1 is 1, Y does not exist and m2 is 0.

[0020] R 3 It is H, methylene or C 1-6 alkyl;

[0021] n2 is 0 or 1;

[0022] A3 is a 5- or 6-membered carbon ring or heterocyclic ring.

[0023] A3 is optionally substituted by 0, 1, 2 or 3 independent substituents selected from the following: OH, halogen, C 1-6 Alkyl, C 1-6 Alkoxy,

[0024] in C 1-6 The alkoxy group may optionally be substituted by 0, 1, or 2 independent substituents selected from the following: halogen, OH, CH3, OCH3, COOH, NH2, NH(C 1-6 alkyl), N(C) 1-6 Alkyl)2,

[0025] Where n3 is 0, 1, 2, 3 or 4.

[0026] Where X is CH2, NH or O;

[0027] When n2 is 0, R 3 It is a methylene group, and the A3 is optionally C 1-6 When alkyl is substituted, the R 3 and the aforementioned C 1-6 Alkyl groups, together with the atoms they are attached to, form 6- to 8-membered heterocycles;

[0028] Or a pharmaceutically acceptable salt, ester, stereoisomer, tautomer, solvate, prodrug, or combination thereof.

[0029] In some embodiments, A3 is optionally substituted by 0, 1, or 2 substituents independently selected from the following:

[0030] OH, methylene, F, Cl, Br, -CH3, -OMe, -OEt, OBn, -OCF3

[0031] In some implementations, A2 is

[0032] In one implementation, n1 is 1;

[0033] A1 is a phenyl group or independently selected from... A1 is optionally substituted by 0 or 1 substituent selected from the following: OH, F, Cl or methyl.

[0034] In a preferred embodiment, m1 is 0, m2 is 1, and Y is C.

[0035] In a more preferred embodiment, n2 is 0;

[0036] A3 is selected from phenyl, pyridyl, cyclohexyl, pyrrolidinyl, or piperidinyl.

[0037] In a preferred embodiment, A3 is selected from pyridinyl, cyclohexyl, pyrrolidinyl, or piperidinyl;

[0038] A3 is optionally substituted by 0, 1, or 2 independent substituents selected from the following: OH, halogen, C. 1-6 Alkyl, C 1-6 alkoxy or In this case, C 1-6 Alkyl groups can be -CH3, and C 1-6 The alkoxy group can be -OMe.

[0039] In an alternative preferred embodiment, A3 is phenyl, wherein A3 is optionally substituted by 0, 1, or 2 substituents independently selected from the group consisting of:

[0040] OH, methylene, F, Cl, Br, -CH3, -OMe, -OEt, OBn, -OCF3

[0041]

[0042] In another preferred embodiment, m1 is 0, m2 is 2, and Y is S;

[0043] n2 is 0;

[0044] A3 is selected from phenyl, pyridyl, cyclohexyl, pyrrolidinyl, or piperidinyl.

[0045] In a more preferred embodiment, A3 is selected from pyridinyl, cyclohexyl, pyrrolidinyl, or piperidinyl;

[0046] A3 is optionally substituted by 0, 1, or 2 independent substituents selected from the following: OH, halogen, C. 1-6 Alkyl, C 1-6 alkoxy or In this case, C 1-6 Alkyl groups can be -CH3, and C 1-6 The alkoxy group can be -OMe.

[0047] In another implementation, n1 is 0;

[0048] A1 is a phenyl group or independently selected from... A1 is optionally substituted by 0 or 1 substituent selected from the following: OH, F, Cl or methyl.

[0049] In a preferred embodiment, m1 is 0, m2 is 1, and Y is C.

[0050] In a more preferred embodiment, n2 is 0;

[0051] A3 is selected from phenyl, pyridyl, cyclohexyl, pyrrolidinyl, or piperidinyl.

[0052] In a preferred embodiment, A3 is selected from pyridinyl, cyclohexyl, pyrrolidinyl, or piperidinyl;

[0053] A3 is optionally substituted by 0, 1, or 2 independent substituents selected from the following: OH, halogen, C. 1-6 Alkyl, C 1-6 alkoxy or In this case, C 1-6 Alkyl groups can be -CH3, and C 1-6 The alkoxy group can be -OMe.

[0054] In an alternative preferred embodiment, m1 is 0, m2 is 2, and Y is S.

[0055] n2 is 0;

[0056] A3 is selected from phenyl, pyridyl, cyclohexyl, pyrrolidinyl, or piperidinyl.

[0057] In a more preferred embodiment, A3 is selected from pyridinyl, cyclohexyl, pyrrolidinyl, or piperidinyl;

[0058] A3 is optionally substituted by 0, 1, or 2 independent substituents selected from the following: OH, halogen, C. 1-6 Alkyl, C 1-6 alkoxy or In this case, C 1-6 Alkyl groups can be -CH3, and C 1-6 The alkoxy group can be -OMe.

[0059] In yet another implementation, n1 is 0 or 1; m1 is 1, Y does not exist, and m2 is 0.

[0060] In a preferred embodiment, n2 is 0.

[0061] A1 is a phenyl group or independently selected from... A1 is optionally substituted by 0 or 1 substituent selected from the following: OH, F, Cl or methyl.

[0062] In a more preferred embodiment,

[0063] A3 is selected from phenyl, pyridyl, cyclohexyl, pyrrolidinyl, or piperidinyl.

[0064] In yet another implementation scheme

[0065] A1 is a phenyl group or independently selected from... A1 is optionally substituted by 0 or 1 substituent selected from the following: OH, F, Cl or methyl;

[0066] n1 is 0 or 1;

[0067] m1 is 0, m2 is 1, and Y is C;

[0068] n2 is 0;

[0069] R 3 It is methylene; A3 is C. 1-6 Alkyl-substituted phenyl; the methylene group and the C 1-6 Alkyl groups, together with the atoms they are attached to, form 6- to 8-membered heterocycles.

[0070] In a preferred embodiment, wherein

[0071] n1 is 1; R 3 It is methylene; A3 is a methyl-substituted phenyl group; and

[0072] yes It may optionally be substituted by 0, 1, or 2 independent substituents selected from the following: halogens,

[0073] In yet another embodiment, the compound is selected from...

[0074]

[0075]

[0076]

[0077]

[0078]

[0079]

[0080]

[0081]

[0082]

[0083] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula I above, or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient.

[0084] In another aspect, the present invention provides the use of the compound or pharmaceutical composition of Formula I above for treating diseases mediated by MIF.

[0085] Preferably, the disease mediated by MIF is

[0086] Tumors selected from glioblastoma, lung cancer, breast cancer, gastric cancer, bladder cancer, and melanoma;

[0087] Inflammatory diseases such as chronic obstructive pulmonary disease (COPD) and pneumonia;

[0088] It is selected from autoimmune diseases such as rheumatoid arthritis (RA), multiple sclerosis (MS), and systemic lupus erythematosus (SLE).

[0089] In another aspect, the present invention provides the use of the compound or pharmaceutical composition of Formula I above for the preparation of a medicament for treating diseases mediated by MIF.

[0090] Preferably, the disease mediated by MIF is

[0091] Tumors selected from glioblastoma, lung cancer, breast cancer, gastric cancer, bladder cancer, and melanoma;

[0092] Inflammatory diseases such as chronic obstructive pulmonary disease (COPD) and pneumonia;

[0093] It is selected from autoimmune diseases such as rheumatoid arthritis (RA), multiple sclerosis (MS), and systemic lupus erythematosus (SLE).

[0094] In another aspect, the present invention provides a method for treating a disease mediated by MIF in a subject in need, comprising administering the subject a therapeutically effective amount of the compound of formula I above, or a pharmaceutically acceptable salt thereof.

[0095] Preferably, the disease mediated by MIF is

[0096] Tumors selected from glioblastoma, lung cancer, breast cancer, gastric cancer, bladder cancer, and melanoma;

[0097] Inflammatory diseases such as chronic obstructive pulmonary disease (COPD) and pneumonia;

[0098] It is selected from autoimmune diseases such as rheumatoid arthritis (RA), multiple sclerosis (MS), and systemic lupus erythematosus (SLE).

[0099] Preferably, the subject is a human.

[0100] In another aspect, the present invention provides a method for inhibiting the expression, production and / or secretion of MIF in a subject in need, the method comprising administering the subject a pharmaceutically effective amount of the compound of formula I above, or a pharmaceutically acceptable salt thereof.

[0101] Preferably, the subject is a human.

[0102] In another aspect, the present invention provides a method for inhibiting the catalytic activity of MIF tautomerase in a subject in need, the method comprising administering the subject a pharmaceutically effective amount of the compound of formula I above, or a pharmaceutically acceptable salt thereof.

[0103] Preferably, the subject is a human.

[0104] In another aspect, the present invention provides a method for inhibiting the expression, production and / or secretion of MIFs in cells, comprising contacting the cells with a pharmaceutically effective amount of a compound of formula I above, or a pharmaceutically acceptable salt thereof.

[0105] In another aspect, the present invention provides a method for inhibiting the catalytic activity of MIF tautomerases in cells, comprising contacting the cells with a pharmaceutically effective amount of the compound of formula I above, or a pharmaceutically acceptable salt thereof. Detailed Implementation

[0106] definition

[0107] In this invention, the following definitions apply:

[0108] The term "MIF inhibitor" includes any and all possible isomers, stereoisomers, enantiomers, diastereomers, tautomers, pharmaceutically acceptable salts, hydrates, solvates, and prodrugs of the MIF inhibitors described in this invention.

[0109] In this invention, the article “a / an” is used to refer to one or more (i.e., at least one) grammatical object of the article. For example, “an element” refers to one element or more elements.

[0110] Unless otherwise stated, the term “and / or” is used in this invention to mean “and” or “or”.

[0111] As used in this invention, the term "carbocyclic ring" refers to a 3-10 membered aromatic or non-aromatic cyclic carbon chain. Examples of non-aromatic carbocyclic rings include cyclopropyl, cyclobutyl, and cyclopentyl. Examples of aromatic carbocyclic rings include benzene, naphthalene, anthracene, and phenanthrene.

[0112] As used in this invention, the term "heterocyclic" refers to a cyclic hydrocarbon containing 3 to 10 atoms, wherein at least one atom is O, N, or S, and a monocyclic heterocyclic may contain up to two double bonds. Examples of heterocyclic compounds include, but are not limited to, aziridine, ethylene oxide, cyclothioethane, azacyclic butane, oxacyclic butane, thiocyclic butane, pyrrolidine, tetrahydrofuran, tetrahydrothiophene, piperidine, tetrahydropyran, thiazolidine, imidazolide, oxazolidine, thiazolide, dioxopentane, dithiocyclic pentane, piperazine, oxazine, dithiazolidine, and dioxane.

[0113] Term "C" 1-6 "Alkyl" refers to a straight-chain or branched alkyl group, alone or in combination with other groups, having 1 to 6 carbon atoms. C 1-6 Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, sec-butyl, tert-butyl, isopentyl, and neopentyl.

[0114] Term "C" 3-6 "Cycloalkyl" refers to a cyclic hydrocarbon containing 3 to 6 carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. It should be understood that any substituted hydrogen atom on a cycloalkyl group can be replaced by a halogen, a C1-C3 alkyl group, a hydroxyl group, an alkoxy group, or a cyano group.

[0115] Term "C" 1-6 "Alkoxy" refers to the group R'-O-, alone or in combination with other groups, where R' is a carbon atom. 1-6 Alkyl group. C 1-6 Examples of alkoxy groups include, but are not limited to, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, tert-butoxy, pentoxy, isopentoxy, neopentoxy, tert-pentoxy, and hexoxy.

[0116] The term "halogen" or "halogen group" refers to fluorine, chlorine, bromine, or iodine. Preferred "halogen" groups are fluorine, chlorine, or bromine.

[0117] "Optional" or "optionally" means that the event or situation described below may or may not occur, and the description includes both the possibility that the event or situation will occur and the possibility that it will not occur.

[0118] The “substituted” group includes embodiments in which a monovalent substituent is bonded to a single atom of the substituted group (e.g., forming a branch), and also embodiments in which the substituent can be a bivalent bridging group bonded to two adjacent atoms of the substituted group, thereby forming a fused ring on the substituted group.

[0119] In this invention, where a given group (partially) is described as being attached to a second group and the attachment site is not specified, the given group can be attached to any available site of the second group at any available site of the given group. For example, "C 1-6 "Alkyl-substituted phenyl", when the linkage site is unclear, can allow C 1-6 Any available site of an alkyl group can be linked to any available site of a phenyl group. In this context, a "available site" is a group site where the hydrogen atoms of the group can be replaced by substituents.

[0120] "Pharmaceutically acceptable salts" are those that retain the biological potency and properties of a free base and are obtained by reaction with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid, or organic acids such as sulfonic acid, carboxylic acid, organophosphoric acid, methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, citric acid, fumaric acid, maleic acid, succinic acid, benzoic acid, salicylic acid, lactic acid, tartaric acid (e.g., (+) or (-)-tartaric acid or mixtures thereof), amino acids (e.g., (+) or (-)-amino acids or mixtures thereof). These salts can be prepared by methods known to those skilled in the art. "Pharmaceutically acceptable salts" are those that, within reasonable medical judgment, are suitable for contact with tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SMBerge et al. described pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, which is incorporated herein by reference.

[0121] The term "carrier" as used in this invention includes carriers, excipients, and diluents, and refers to materials, compositions, or excipients, such as liquid or solid fillers, diluents, excipients, solvents, or encapsulating materials, that participate in the transport or delivery of a pharmaceutical agent from one organ or part of the body to another organ or part of the body.

[0122] The term "effective amount" refers to the amount of the compound of the present invention that is sufficient to achieve the desired effect or result when it is given or used. As appropriate, the term effective amount may include pharmaceutically effective amount or therapeutically effective amount or synonyms thereof. Effective amount can be determined by methods known to those skilled in the art.

[0123] Compounds of the given formula (e.g., compounds of formula I) are intended to include pharmaceutically acceptable salts, pharmaceutically acceptable esters, isomers, tautomers, solvates, isotopes, hydrates, polymorphs, and prodrugs of the compounds of the present invention. Furthermore, compounds of the present invention may have one or more asymmetric centers and may be produced as racemic mixtures or as individual enantiomers or diastereomers. The number of stereoisomers present in any given compound of the given formula depends on the number of asymmetric centers present (possibly 2). n There are n stereoisomers, where n is the number of asymmetric centers. Individual stereoisomers can be obtained by resolving racemic or non-racemic mixtures of intermediates at appropriate stages of synthesis or by resolving the compound using conventional methods.

[0124] Individual stereoisomers (including individual enantiomers and diastereomers) as well as racemic and non-racemic mixtures of stereoisomers are all included within the scope of this invention, and all of these are intended to be described by the structure of this specification unless otherwise specifically stated.

[0125] "Isomers" are different compounds that have the same molecular formula. Isomers include stereoisomers, enantiomers, and diastereomers.

[0126] "Stereoisomers" are isomers that differ only in the spatial arrangement of atoms.

[0127] An enantiomer is a pair of stereoisomers that are non-overlapping mirror images of each other. A 1:1 mixture of a pair of enantiomers is a racemic mixture. Where appropriate, the term "(±)" is used to denote a racemic mixture.

[0128] A “diastereomer” is a stereoisomer that has at least two asymmetric atoms that are not mirror images of each other.

[0129] Some compounds exist as tautomers. These tautomers exist in equilibrium with each other. For example, amide-containing compounds can exist in equilibrium with their imine tautomers. Regardless of the type of tautomer exhibited or the nature of the equilibrium between the tautomers, those skilled in the art will understand that these compounds include both amide and imine tautomers. Therefore, amide-containing compounds should be understood to include their imine tautomers. Similarly, imine-containing compounds should be understood to include their amide tautomers.

[0130] The term "polymorph" refers to different crystal structures of a crystalline compound. Different polymorphs may be due to differences in crystal packing (packing polymorphism) or differences in packing between different conformational isomers of the same molecule (conformational polymorphism).

[0131] The term "solvent" refers to a complex formed by combining the compounds of the present invention with a solvent.

[0132] The term "hydrate" refers to a complex formed by combining the compounds of the present invention with water.

[0133] The term "prodrug" refers to the compounds of the present invention, pharmaceutically acceptable salts thereof, or biologically active metabolites thereof that include chemical groups that can be converted in vivo from the remainder of the molecule and / or isolated from the remainder of the molecule to provide an active medicament.

[0134] "Subject" is a mammal, such as a human, mouse, rat, guinea pig, dog, cat, horse, cow, pig, or a non-human primate, such as a monkey, chimpanzee, baboon, or rhesus monkey, and the terms "subject" and "patient" are used interchangeably in this invention.

[0135] The term "inhibition" refers to a significant reduction in the baseline activity of a biological activity or process.

[0136] Unless otherwise stated, the term "disease" is used in this invention to refer to the terms symptom, condition, or ailment, and may be used interchangeably with these terms.

[0137] The term "tumor" as used in this invention refers to the abnormal growth of tissue. Tumors can be benign or malignant. Generally, malignant tumors are referred to as cancer. Cancer differs from benign tumors in the ability of malignant cells to invade other tissues, either by growing directly into adjacent tissues or by metastasizing (i.e., by transporting via the blood or lymphatic system) to distant sites.

[0138] As used in this invention, the term "autoimmune disease" refers to a disease caused by an inappropriate immune response of the body to substances and tissues normally present in the patient's body (autoimmunity). Symptoms of autoimmune diseases range from fatigue and mild skin rashes to rare but severe warning signs, such as seizures. Autoimmune diseases are preferably selected from rheumatoid arthritis (RA), multiple sclerosis (MS), sarcoidosis, psoriasis, Crohn's disease, systemic lupus erythematosus (SLE), and type 1 diabetes, and more preferably from rheumatoid arthritis (RA), multiple sclerosis (MS), and systemic lupus erythematosus (SLE).

[0139] As used in this invention, the term "inflammatory disease" refers to a disease caused by, resulting from, or leading to inflammation. The term "inflammatory disease" can also refer to a dysregulated inflammatory response that causes an excessive response of macrophages, granulocytes, and / or T lymphocytes, resulting in abnormal tissue damage and / or cell death. Inflammatory diseases can be acute or chronic inflammatory conditions and can be caused by infectious or non-infectious factors. Chronic obstructive pulmonary disease (COPD) and pneumonia are preferred examples of inflammatory diseases.

[0140] The term "treatment," in the context of a subject, refers to the improvement of at least one symptom of the subject's disease. Treatment can be curative, improvement, or at least partial relief of the disease.

[0141] The term "administer / administering / administration" as used in this invention refers to the direct administration of a compound or a pharmaceutically acceptable salt or composition of a compound to a subject, or the administration of a prodrug derivative or analog of a compound or a pharmaceutically acceptable salt or composition of a compound to a subject, which can form an equivalent amount of the active compound in the subject's body.

[0142] Pharmaceutical Composition and Administration

[0143] The present invention also includes pharmaceutical compositions for treating diseases mediated by MIFs, or for inhibiting MIF expression, production and / or secretion, or for inhibiting MIF tautomerase catalytic activity, or more than one of these activities. The compositions may be suitable for internal use and comprise an effective amount of the compound of the present invention as a MIF inhibitor and a pharmaceutically acceptable carrier. MIF inhibitors are particularly useful because they exhibit very low or no systemic toxicity.

[0144] MIF inhibitors may each be administered in an amount sufficient to treat or prevent, but not limited to, cardiovascular and cerebrovascular diseases, autoimmune diseases and inflammatory conditions, fibrotic diseases, metabolic diseases and tumors, or to prevent their development, in subjects.

[0145] MIF inhibitors can be administered via any of the therapeutic routes. These routes include systemic or local administration, such as oral, nasal, parenteral (intravenous), intramuscular, intrathecal, intravitreal, transdermal, subcutaneous, vaginal, oral, rectal, local administration, or as a drug-eluting stent.

[0146] Depending on the intended route of administration, the compositions can be in solid, semi-solid, or liquid dosage forms, such as injections, tablets, suppositories, pills, sustained-release capsules, elixirs, tinctures, emulsions, syrups, powders, liquids, suspensions, etc., sometimes in unit doses and in accordance with conventional pharmaceutical practice. Similarly, they can be administered intravenously (bottle and infusion), intraperitoneally, intrathecally, intravitreally, subcutaneously, or intramuscularly, all using forms familiar to those skilled in the art of pharmaceutical work.

[0147] The illustrative pharmaceutical compositions are tablets and gelatin capsules containing a MIF inhibitor and a pharmaceutically acceptable carrier, such as: a) diluents, such as purified water, triglyceride oils such as hydrogenated or partially hydrogenated vegetable oils or mixtures thereof, corn oil, olive oil, sunflower oil, safflower oil, fish oils such as EPA or DHA or their esters or triglycerides or mixtures thereof, omega-3 fatty acids or their derivatives, lactose, dextran, sucrose, mannitol, sorbitol, cellulose, sodium, saccharin, glucose and / or glycine; b) lubricants, such as silica, talc, stearic acid, its magnesium or calcium salts, sodium oleate, sodium stearate, magnesium stearate. Sodium benzoate, sodium acetate, sodium chloride, and / or polyethylene glycol; for tablets, it also contains c) binders, such as magnesium aluminum silicate, starch paste, gelatin, tragacanth gum, methylcellulose, sodium carboxymethyl cellulose, magnesium carbonate, natural sugars such as glucose or β-lactose, corn sweeteners, natural and synthetic gums such as gum arabic, tragacanth, or sodium alginate, waxes, and / or polyvinylpyrrolidone (if applicable); d) disintegrants, such as starch, agar, methylcellulose, bentonite, xanthan gum, alginate or its sodium salt, or effervescent mixtures; e) absorbents, colorants, flavoring agents, and sweeteners; f) emulsifiers or dispersants, such as Tween 80. Labrasol, HPMC, DOSS, caproyl 909, labrafac, labrafil, peceol, transcutol, capmul MCM, capmul PG-12, captex 355, gelucire, vitamin E TGPS or other acceptable emulsifiers; and / or g) agents that enhance the absorption of the compound, such as cyclodextrin, hydroxypropyl-cyclodextrin, PEG400, PEG200.

[0148] Liquids, particularly injectable compositions, can be prepared, for example, by dissolution, dispersion, etc. For instance, MIF inhibitors can be dissolved in or mixed with pharmaceutically acceptable solvents such as water, saline, dextran aqueous solution, glycerol, ethanol, etc., to form injectable isotonic solutions or suspensions. Proteins such as albumin, chylomicrons, or serum proteins can be used to dissolve MIF inhibitors.

[0149] MIF inhibitors can also be formulated into suppositories, which can be prepared from fat emulsions or suspensions; using polyalkylene glycols such as propylene glycol as carriers.

[0150] In other embodiments, the pharmaceutical formulations described in this invention include, but are not limited to, aqueous liquid dispersions, self-emulsifying dispersions, solid solutions, liposome dispersions, aerosols, solid dosage forms, powders, immediate-release formulations, controlled-release formulations, fast-dissolving formulations, delayed-release formulations, extended-release formulations, pulsatile-release formulations, multi-particle formulations, and mixed immediate-release and controlled-release formulations.

[0151] MIF inhibitors can also be administered in the form of liposome delivery systems, such as small unilamellar vesicles, large unilamellar vesicles, and multilayer vesicles. Liposomes can be formed from a variety of phospholipids, including cholesterol, stearamine, or phosphatidylcholine. In some embodiments, such as those described in U.S. Patent No. 5,262,564, the lipid component membrane is hydrated with an aqueous drug solution to form a lipid layer encapsulating the drug.

[0152] MIF inhibitors can also be delivered using monoclonal antibodies as separate carriers conjugated to the MIF inhibitors. MIF inhibitors can also be conjugated to soluble polymers that serve as targeted drug carriers. Such polymers can include polyvinylpyrrolidone, pyran copolymers, polyhydroxypropyl methacrylamide-phenol, polyhydroxyethyl asparagine-phenol, or poly(ethylene oxide) polylysine substituted with palmitoyl residues. Furthermore, MIF inhibitors can be conjugated to a class of biodegradable polymers that can be used to achieve controlled drug release, such as crosslinked or amphiphilic block copolymers of polylactic acid, poly(ε-caprolactone), polyhydroxybutyric acid, polyorthoesters, polyacetals, polydihydropyran, polycyanoacrylates, and hydrogels. In one embodiment, the MIF inhibitor is not covalently bound to a polymer, such as a polycarboxylic acid polymer or a polyacrylate.

[0153] Parenteral administration is typically performed via subcutaneous, intramuscular, or intravenous injection and infusion. Injectable preparations can be prepared in conventional forms, such as liquid solutions or suspensions, or in solid forms suitable for dissolving in a liquid prior to injection.

[0154] The composition can be prepared according to conventional mixing, granulation or coating methods, and the pharmaceutical composition may contain about 0.1% to about 80%, about 5% to about 60%, or about 1% to about 20% of MIF inhibitor by weight or volume.

[0155] The dosing regimen for MIF inhibitors is chosen based on a variety of factors, including patient type, species, age, weight, sex, ethnicity, diet, and medical condition; the severity of the condition to be treated; route of administration; the patient's renal or hepatic function; and the specific MIF inhibitor used. Physicians or veterinarians with ordinary skills in the art can readily determine and prescribe the effective amount of medication needed to prevent, counteract, or halt the progression of the condition.

[0156] The effective dosage of the present invention, when used for the specified effect, can be from about 0.1 mg to about 5000 mg of the active ingredient per unit dose. In one embodiment, the composition is in the form of a scoreable tablet. The appropriate dosage of the MIF inhibitor can be determined as described in Goodman, LS; Gilman, A. The Pharmacological Basis of Therapeutics, 5th ed.; MacMillan: New York, 1975, pp. 201-226.

[0157] MIF inhibitors can be administered as a single daily dose, or the total daily dose can be administered in divided doses two, three, or four times daily. Alternatively, MIF inhibitors can be administered intranasally using a suitable intranasal medium, or via a transdermal route using transdermal skin patches of the forms known to those skilled in the art. For administration via a transdermal delivery system, dosing can be continuous rather than intermittent throughout the dosing regimen. Other illustrative topical formulations include creams, ointments, lotions, aerosol sprays, and gels, wherein the concentration range of the MIF inhibitor is from about 0.1% to about 15% w / w or w / v.

[0158] Uses of compounds and their compositions

[0159] The compounds and compositions described above as MIF inhibitors can be used to treat or prevent MIF-related diseases. These diseases include, but are not limited to, autoimmune diseases, tumors, or chronic or acute inflammatory diseases. Examples of such diseases or conditions include:

[0160] ο Rheumatic diseases (including but not limited to rheumatoid arthritis, osteoarthritis, psoriatic arthritis), spondyloarthritis (including but not limited to ankylosing spondylitis, reactive arthritis, Reiter's syndrome), crystal arthropathy (including but not limited to gout, pseudogout, calcium pyrophosphate deposition disease), Lyme disease, polymyalgia rheumatica;

[0161] Connective tissue diseases (including but not limited to systemic lupus erythematosus, systemic sclerosis, polymyositis, dermatomyositis, Sjögren's syndrome) syndrome);

[0162] ο Vasculitis (including but not limited to polyarteritis nodosa, Wegener's granulomatosis, and Churg-Strauss syndrome);

[0163] Inflammatory diseases, including chronic obstructive pulmonary disease (COPD), pneumonia, and the consequences of trauma or ischemia;

[0164] Sarcoidosis;

[0165] Vascular diseases, including atherosclerotic vascular diseases and infarctions, atherosclerotic and occlusive vascular diseases (including but not limited to atherosclerosis, ischemic heart disease, myocardial infarction, stroke, peripheral vascular disease) and vascular stent restenosis;

[0166] Autoimmune diseases (including but not limited to diabetes, thyroiditis, myasthenia gravis, sclerosing cholangitis, primary biliary cirrhosis, rheumatoid arthritis (RA), multiple sclerosis (MS), and systemic lupus erythematosus (SLE));

[0167] Lung diseases (including but not limited to diffuse interstitial lung disease, pneumoconiosis, fibrotic alveolitis, asthma, bronchitis, bronchiectasis, chronic obstructive pulmonary disease, and adult respiratory distress syndrome);

[0168] Primary or metastatic tumors (including but not limited to glioblastoma, prostate cancer, colon cancer, lymphoma, lung cancer, gastric cancer, bladder cancer, melanoma, multiple myeloma, breast cancer, gastric cancer, leukemia, cervical cancer and metastatic cancer);

[0169] Nephropathy, including glomerulonephritis and interstitial nephritis;

[0170] Hypothalamic-pituitary-adrenal axis disorders;

[0171] Neurological disorders, including multiple sclerosis and Alzheimer's disease;

[0172] Diseases characterized by altered angiogenesis (e.g., diabetic retinopathy, rheumatoid arthritis, cancer) and endometrial function (menstruation, implantation, endometriosis);

[0173] Complications of infectious diseases, including endotoxin (septic) shock, exotoxin (septic) shock, septic (true septic) shock, malaria complications, other infectious complications, and pelvic inflammatory disease;

[0174] Transplant rejection, graft-versus-host disease;

[0175] Allergic diseases, including allergies, atopic diseases, and allergic rhinitis;

[0176] ο Bone diseases (such as osteoporosis, Paget's disease);

[0177] Skin diseases, including psoriasis, atopic dermatitis, and UV(B)-induced dermal cell activation (e.g., sunburn, skin cancer);

[0178] Pain, testicular dysfunction, and wound healing;

[0179] Gastrointestinal diseases, including inflammatory bowel disease (including but not limited to ulcerative colitis and Crohn's disease), peptic ulcers, gastritis, esophagitis, and liver diseases (including but not limited to cirrhosis and hepatitis).

[0180] Combination therapy

[0181] The compounds of this invention can be administered in combination with the following non-limiting examples of therapeutic agents and methods, for the treatment and prevention of these MIF-related diseases, including but not limited to any combination of the following: glucocorticoids, nonsteroidal anti-inflammatory drugs (NSAIDs), cyclooxygenase (COX)-2 inhibitors, licofelone (ML3000), disease-modifying antirheumatic drugs (DMARDs), methotrexate, chloroquine, hydroxychloroquine, cyclophosphamide (Cytoxan), inosine monophosphate dehydrogenase (IMPDH) inhibitors, sirolimus, everolimus (rapamycin), purine nucleoside phosphorylase inhibitors, purine de novo synthesis inhibitors, and dihydroorotate dehydrogenase inhibitors (malononitrile amide). Prostaglandin PGE2 inhibitors, P2X7 receptor inhibitors, protease-activated receptor 2 (PAR-2) inhibitors, complement activation inhibitors, complement C3 / C5 convertase inhibitors, active convertase inhibitors, complement C5aR antagonists, EP4 agonists, prostaglandin I2 analogs, sulfasalazine (SASP), 5-aminosalicylic acid (5-ASA), immunomodulatory drugs, calcineurin inhibitors, interleukin-10 (AG011), placental-derived cells (PDA001), mucosal addressing cell adhesion molecule (MAdCAM) inhibitors (PF-00547659), GLP-2 agonists, anti-CD3, CCR9 inhibitors, lenalidomide Relimid, recombinant human interleukin-11, CXCR2 antagonists, glucagon-like peptide-2 (GLP-2) analogs (teduglutide), insulin-like growth factor-1 (IGF-1) (Increlex), synthetic amitrazine sermamod (CPSI-2364), intracellular adhesion molecule-1 (ICAM-1) inhibitors (alicaforsen), stem cell therapy, activated protein C (aPC), vitamin D analogs, visual pigments, phototherapy, methotrexate, cyclosporine, acitretin, CCR6 inhibitors, CCL20 inhibitors, deoxyguanidin, alkylated deoxyribonucleic acid (DNA) agents, Tumor necrosis factor (TNF)-α inhibitors, TNF-α converting enzyme inhibitors, Janus kinase (JAK) 1, 2 and / or 3 inhibitors, spleen tyrosine kinase (SYK) inhibitors, caspase inhibitors, chemokine receptor antagonists, protein kinase C (PKC) inhibitors, p38 mitogen-activated protein kinase (MAPK) inhibitors, caspase inhibitors, NF-κB modulators, B cell inhibitors, hydroxychloroquine, B lymphocyte stimulator (BLyS) inhibitors, membrane-bound and soluble B cell activating factor inhibitors, inhibitors that antagonize BLyS and APRIL (proliferation-inducing ligand) cytokines to prevent B cell maturation and autoantibody production, anti-CD19, CD20 inhibitors, CD22 inhibitors, T cell inhibitors, interferon inhibitors.Toll-like receptor inhibitors, praserotonin, estrogen receptor antagonists (fulvestrant), cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4)-Ig, V-set domain-containing T-cell activation inhibitor 1 (VTCN1; B7-H4) agonist (AMP-110), interleukin-1 receptor antagonists (AMG) 108, anakinetin [KINERET], interleukin-1β antagonists, soluble IL-1 receptors, interleukin-2R antagonists, interleukin-6 receptor antagonists, calcipotriol / betamethasone (Taclonex), fumarate (Panaclar / BG-12), interleukin-15 inhibitors, interleukin-17 inhibitors (AIN457), DHODH inhibitors (Vidofludimus), interleukin-18 inhibitors, T helper (Th)17 cell inhibitors, interleukin-12 / interleukin-23 inhibitors, interleukin-22 inhibitors, interleukin-23 inhibitors, interleukin-12 inhibitors, alpha interferon β-interferon [interferon β-1a (Avonex, Rebif), interferon β-1b (Betaseron / Betaferon), glatiramer acetate (Copaxone), selective adhesion molecule inhibitors, integrin antagonists (natezumab [Tysabri], vedolizumab), sphingosine 1-phosphate receptor (S1P-R) agonists, fumarate derivative immunomodulators, lacquimod, anti-LFA-1, MBP-8298, cladribine, Novantrone, isoxanol dihydroorotate dehydrogenase (DHODH) and tyrosine kinase inhibitors, Revimmune (cyclophosphamide), fampridine SR (4-aminopyridine), Panaclar (dimethyl fumarate), MBP8298 (dirucotide, a synthetic peptide version of some human myelin basic protein), Camppath (alemumab), anti-CD52, purine analogs, fingolimod (sphingosine 1-phosphate receptor agonist), teriflunomide, pyrimidine de novo synthesis inhibitors, leflunomide's active metabolite, verteporfen photodynamic therapy [PDT], anti-angiogenic factors, CCR3 inhibitors, anti-CD48, β2-agonists, leukotriene modulators Phosphodiesterase (PDE) inhibitors, selective phosphodiesterase-4 (PDE-4) inhibitors, IgE-targeting inhibitors (omalizumab), Th2 cytokine inhibitors, macrolides, ketolides, adenosine A2B antagonists, κB kinase 2 inhibitors, prostaglandin-like and F2-isoprostaglandin antagonists, nitric oxide donors, inducible nitric oxide synthase inhibitors, Toll-like receptor modulators, lorcaserin, phentermine, topiramate, bupropion, naltrexone, anti-thymocyte globulin, serine protease inhibitors, tyrosine kinase inhibitors.Platelet-derived growth factor receptor (PDGFR) inhibitors, insulin, type 1 diabetes antigen-specific tolerance inducers, cannabinoid receptor 1 (CB1) antagonists, long-acting glucagon-like peptide-1 (GLP1) analogs, dipeptidyl peptidase-4 (DPP4) inhibitors, vasoactive intestinal peptide-pituitary adenylate cyclase-activated peptide receptor 2 (VPAC2) agonists, glucokinase activators, glucagon receptor antagonists, cytosolic phosphoenolpyruvate carboxykinase (PEPCK) inhibitors, sodium-glucose cotransporter 2 (SGLT2) inhibitors, salsalate, IκB kinase-β (IKKβ) inhibitors, nuclear factor κB inhibitors, interleukin-1 (IL-1) receptor antagonists, IL-1β specific antibodies, sirtuin SIRT1 activators, selective peroxisome proliferator-activated receptor (PPAR) modulators (SPPARM), 11β-hydroxysteroid dehydrogenase type 1 (11βHSD1) inhibitors, PPARγ ligands, thiazolidinediones, glitazones, warfarin, coumarin, pradaxa, antithrombotic drugs, statins, HMG-CoA reductase inhibitors, ezetimibe, fenofibrate, niacin, amlodipine, vascular cell adhesion molecule (VCAM) antagonists, thromboxane A2 antagonists, prostaglandin D2 receptor 1 antagonists, G protein-coupled receptor (GPCR) modulators, cannabinoid receptor 1 (also known as CNR1) CB1 receptor antagonists (rimonaban), cholesterol ester transfer protein (CETP) inhibitors (JTT-705), chemokine (CC motif) receptor 2 (CCR2) antagonists, phospholipase A2 inhibitors, peroxisomes Proliferator-activated receptor (PPAR) agonists, RNA polymerase inhibitors, leukotriene synthesis inhibitors, α7 nicotinic receptor (α7nAChR) agonists, donepezil, galantamine, rivastigmine, memantine, α-secretase cleavage stimulants, γ-secretase activity inhibitors, antioxidant therapy, estrogen, NO synthase inhibitors, anti-β-amyloid (Aβ) (bapinotumab), abiraterone, ActRIIA signaling inhibitor (ACE-011), doxorubicin, interleukin [Proleukin], alemtuzumab, retinoic acid, alkylating agents and microtubule inhibitors, allopurinol, allosteric Akt inhibitor (Akti) [MK-2206], hexamethylmelamine, amifostine, anastrozole (Arimidex), triple angiokinase inhibitors (BIBF) inhibiting VEGF receptors (VEGFR) 1, 2, and 3, fibroblast growth factor receptor, and platelet-derived growth factor receptor. 1120), angiopoietin 1 / 2 neutralizing peptide (AMG 386), anthracycline (amrubicin), antigen-specific cancer immunotherapy drugs (ASCI), antimetabolites (raltitrexed), and Apaziquone (EOquin).Aprepitant, aromatase inhibitors, arsenic trioxide, asparaginase, anaplastic lymphoma kinase (ALK) inhibitors (crizotinib, AP26113), azacytidine (Vidaza), BCG Live, Bcl-2 family inhibitors, Bcr-Abl inhibitors, bendamustine, bexarotine capsules, bexarotine gel, bleomycin, BRAF signaling inhibitors, busulfan intravenous injection, busulfan oral administration, cabazitaxel (Jevtana), cascarone, capecitabine (Xeloda), carboplatin, carmustine, carmustine with Polifeprosan 20 implants, caspase inhibitors, anti-CD23, anti-CD30, anti-CD32, anti-CD33, anti-CD40, chlorambucil, cisplatin, c-Met receptor tyrosine kinase inhibitor (ARQ197), clolarabine (Clolar), CS1 inhibitors, cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4) inhibitors, cyclophosphamide, cytarabine, cytarabine liposomes, dacarbazine, cytosine, actinomycin D, alpha-dabepoetine (Ar anesp), daunorubicin liposomes, daunorubicin, daunorubicin, histone deacetylase (HDAC) inhibitors, decitabine (Dacogen), delta-like 4-ligand (DLL4) inhibitors (OMP-21M18), denileukin, diftitox, dexrazoxan, docetaxel (Taxotere), doxorubicin, doxorubicin liposomes, drotalodone propionate, DR5 agonist (LBY135), Elliott's Solution B, Epidermal Growth Factor Receptor (EGFR) Inhibitor, Epidermal Growth Factor Receptor Tyrosine Kinase (EGFR-TK) Inhibitor, EGFR Inhibitor-Protein-Tyrosine Kinase Inhibitor, Dual EGFR / HER2 Receptor Tyrosine Kinase Inhibitor, Exalacine, Endothelin-B Receptor Agonist, Epirubicin, Epoetin α and β, Etoposide Phosphate, Etoposide (VP-16), Exemestane, Farnesyltransferase Inhibitor (FTI), Fentanyl, Fluorouracil (Intra-arterial), Fludarabine, Fluorouracil (5-FU), Fulvestrant (Faslodex), G2 Checkpoint Inhibitor (CBP501), GA101, Gemcitabine (Ge mzar), gemutuzumab ozomicin, gonadotropin-releasing hormone (GnRH) agonists, goserelin acetate, granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor (GM-CSF) (Sargramostim), heat shock protein 90 (Hsp90) inhibitors, hedgehog pathway inhibitors (RG3616), pan-HER inhibitors (PF-00299804), Herceptin, HPV vaccine, human death receptor 5 agonists, hydroxyurea, teimiramab (Zevalin), idarubicin, ifosfamide, imatinib mesylate (Gleevec / Glivec), immunomodulatory drugs (IMiD).Insulin-1 receptor (IGF-1R) inhibitors, dual kinase inhibitors of insulin-1 receptor (IGF-1R) and insulin receptor (IR) (OSI-906), interleukin-2, ipilimumab, irinotecan, Istodax (romidesin), lapatinib (Tykerb), leptinib, levofloxacin, levamisole, levofloxacin, LOddC, lomustine (CCNU), leuprolide, luteinizing hormone-releasing hormone (LHRH) agonists and antagonists, thiamethoxam, MAGE-A3 inhibitors, MAPK / ERK kinase 1 / 2 inhibitors (AZD6244), nitrogen mustard (meclorethamine / nitrogen) Mustard, megestrol acetate, melphalan (L-PAM), mercaptopurine (6-MP), mesna, MET inhibitors (XL184), methotrexate, methoxsalen, midotulin (PKC412), mifamotide (Mepact), mitomycin C, mitotane, mitoxantrone, mammalian target of rapamycin (mTOR) inhibitors, MEK inhibitors, microtubule inhibitors, microtubule stabilizers (pertopyron [EP0906]), multi-kinase inhibitors, multi-target receptor tyrosine kinase inhibitors (TKIs) (TKI258), nandrolone phenylpropionate, necitumumab, Neulasta, NK-1 receptor inhibitor, nofetumomab, noscapine (CB3304), ondansetron, oprelvekin, oxaliplatin, PI3K inhibitor, dual PI3K / mTOR inhibitor (BEZ235), paclitaxel, pamidronate Salt, platelet-derived growth factor receptor α (PDGFR-α) inhibitor (IMC-3G3), pegaspargase, pegfilgrastim, pentostatin, pertuzumab, piperobromidine, procainox, Paulo-like kinase 1 (Plk-1) inhibitor, photomycin, poly(ADP-ribose) polymerase-1 (PARP1) inhibitor, porphyrin sodium, integrin avβ3 and avβ5 inhibitors (silengiptide [EMD121974]), pemetrexed (Alimta), pralatrexate injection (Folotyn), praxavir, dual pro-apoptotic receptor (PARA) DR4 and DR5 agonists, procarbazine, protein-tyrosine kinase inhibitors, proteasome inhibitors, quinacrine, raf and VEGFR inhibitors, receptor activators of nuclear factor-κB ligand (RANKL) inhibitors, raburicase, multi-target receptor tyrosine kinase (RTK) inhibitors, romidixin (Istodax), ciocallifloxacin (CB1089), polyethylene glycol-SN38 conjugate (EZN-2208), saplatin,Dual Src and Bcr-Abl kinase inhibitors, streptozotocin, tabuviridine (LDT), talc, tamoxifen (Nolvadex), T-DM1, temozolomide, teniposide (VM-26), testosterone, therapeutic vaccines (BiovaxID), IRX-2, Rindopepimut (CDX-110), sipuleucel-T [Provenge], TVA immunotherapy, Stimuvax [BLP25 liposomal vaccine], somatostatin analogs, taxanes (Ortataxel) Tassolan, Thalidomide, Thioguanine (6-TG), Thiotepa, Topoisomerase I and II inhibitors, Topoisomerase I inhibitors (Gematocan [LBQ707], Irinotecan), Topotecan (Hycamtin), Toremifene, Trabettetin (Yondelis), Trastuzumab, Retinoic acid (ATRA), Tosimozab (Bexxar), TRPM8 agonist (D-3263), Uracil mustard, Recombinant uricase (Elitek), Penrubicin, Vatopalbin (monoval) LDC), antagonists of vascular endothelial growth factor receptors 1, 2, and 3 ("VEGFR1-3") / platelet-derived growth factor receptor ("PDGFR") / stem cell factor receptor ("c-kit") (modexenine), tyrosine kinase inhibitors of vascular endothelial growth factor receptor (VEGFR) / epidermal growth factor receptor (EGFR) / rearrangement (RET) during transfection (vandetanib), vascular endothelial growth factor (VEGF) inhibitors (cidinibub, ramucirumab), VEGFR / EGFR / HER-2 inhibitors (AEE788), vincristine, vinorelbine, Wnt signaling inhibitors (OMP-18R5), zoledronic acid, zoledronic acid and combinations thereof, etc.

[0182] Universal synthesis

[0183] Typical embodiments of the compounds according to the invention can be synthesized using the general reaction scheme described below. As will be apparent from the description herein, the general scheme can be modified by substituting the starting materials with other substances having similar structures to produce correspondingly different products. The description of the synthesis subsequently provides numerous examples of how the starting materials can be varied to yield the corresponding products. Starting materials are typically obtained from commercial sources or synthesized using publicly available methods. The compounds of the invention can be prepared from commercial starting materials using general synthetic techniques and procedures known to those skilled in the art. Reaction schemes suitable for preparing such compounds are outlined below. Further examples can be found in the detailed embodiments described below.

[0184] The compounds disclosed in this invention may have a chiral center, such as a chiral carbon atom. Therefore, such compounds comprise racemic mixtures of all stereoisomers, including enantiomers, diastereomers, and transisomers. Additionally, the compounds disclosed in this invention include optical isomers enriched or resolved at any or all asymmetric chiral atoms. In other words, the chiral center, as apparent from the description, is provided as a chiral isomer or a racemic mixture. Racemic and diastereomer mixtures, as well as isolated or synthesized individual optical isomers substantially free of their enantiomers or diastereomer conjugates, are all within the scope of this invention. Racemic mixtures can be isolated into their respective, substantially optically pure isomers using well-known techniques, such as isolating diastereomers formed with optically active auxiliaries (e.g., acids or bases) and then converting them back to the optically active substance. The desired optical isomers can also be synthesized via stereospecific reactions, starting from suitable stereoisomers of the desired starting material.

[0185] Option 1

[0186]

[0187] In scheme 1, amino-oxadiazole I-2 Prepared via a single-step procedure. In some embodiments, A1 is a phenyl or heterocyclic compound. Amines and carboxylic acids I-3 condensation I-4 In some embodiments, the amide is converted to N-methylamide using a suitable base and alkyl halide under appropriate conditions (such as time and temperature). A1 and R 3 Same as described in this invention, and R 5 And / or Z can be the same as described in this invention regarding the substituents on A3 (i.e., the phenyl in this case).

[0188] Option 2

[0189]

[0190] In scheme 2, methylamino-oxadiazole II-4 It is prepared by a two-step process. This involves making the intermediate... II-3 Intramolecular cyclization followed by removal of the tert-butyloxycarbonyl protecting group yields methylamine. II-4 Amines and carboxylic acids II-5 condensation II-6 In some implementations, under appropriate conditions (such as time and temperature), a suitable base (such as NaH) and an alkyl halide (such as R) are used. 3 I)will II-6 Transform into II-7 A1, R 1 R 2 and R 3The same as described in this invention, and Z can be the same as described in this invention regarding the substituents on A3 (i.e., phenyl in this case).

[0191] Option 3

[0192]

[0193] In scheme 3, Z is a substituent on A3 (i.e., the phenyl group in this case) and can be introduced later using this method. In some embodiments, Z is linked to a nitrogen atom. In this case, the amine Z-NH2 is... III-1a The product is obtained through a palladium-catalyzed reaction. In some embodiments, Z is bonded to a carbon atom, and boric acid ZB(OH)₂ reacts with... III-1a The palladium-catalyzed reaction yields an olefin. The olefin is then reduced with hydrogen to provide the product. Alternatively, the same transformation can be achieved using an organozinc reagent as a coupling complex to provide the product. In some embodiments, Z is bonded to an oxygen atom. III-1b The product is obtained by alkylation with an alkyl halide (such as a Z-halo). In some embodiments, Z is linked to a linking group such as a CH2 unit. The linking group is formed by an amine (such as piperidine, W=CH2) and... III-1c The product was prepared by reductive amination. A1, n1, R 1 and R 2 The same as described in this invention, and Z can be the same as described in this invention regarding the substituents on A3 (i.e., phenyl in this case).

[0194] Option 4

[0195]

[0196] In some implementation schemes using pyridazine, IV-1 With acyl chloride IV-2 Condensation, and then obtained IV-3 The product was obtained by palladium-catalyzed reaction. IV-4 A1 is the same as described in this invention, and Z may be the same as described in this invention regarding the substituents on A3 (i.e., phenyl in this case).

[0197] Option 5

[0198]

[0199] In some implementations using cycloamides, the corresponding chloromethyloxadiazole V-3 Through two steps, V-1 and V-2 R is prepared by starting with a molecular cyclization process. 5 Initially in V-4 superior.V-4 and V-3 Coupling yields products V- 5 A1 is the same as described in this invention, and R 5 This can be the same as the description of the substituents on A3 (i.e., the phenyl in this case) in this invention.

[0200] Example

[0201] The present invention can be better understood from the following embodiments. However, those skilled in the art will readily understand that the descriptions in the embodiments are for illustrative purposes only and are not intended to limit the scope of the invention as detailed in the claims.

[0202] Unless otherwise stated, the compounds of this invention can be prepared from commercial starting materials using common synthetic techniques and procedures known to those skilled in the art. Chromatographic consumables and equipment can be purchased from companies such as: AnaLogix, Inc., Burlington, WI; Analytical Sales and Services, Inc., Pompton Plains, NJ; Teledyne Isco, Lincoln, NE; VWR International, Bridgeport, NJ; and Rainin Instrument Company, Woburn, MA. Chemicals and reagents can be purchased from companies such as: Aldrich, Argonaut Technologies, VWR and Lancaster, Invitrogen, Sigma, Promega, Solarbio, Cisbio, Signalchem, MCE. Consumables can be purchased from companies such as: Corning, Labcyte, Greiner, Nunc. Instruments can be purchased from companies such as: Labcyte, PerkinElmer, Eppendorf, ThermoFisher.

[0203] Example 1

[0204]

[0205] 2-Chloro-N-(5-(furan-2-yl)-1,3,4-oxadiazol-2-yl)-4-methoxybenzamide

[0206] 2-Chloro-4-methoxybenzoyl chloride (369 mg, 1.8 mmol) was added to a solution of 5-(2-furanyl)-1,3,4-oxadiazol-2-amine (90 mg, 0.6 mmol) and DMAP (7 mg, 0.06 mmol) in pyridine (5.0 mL) at 10 °C. The reaction mixture was stirred overnight at 30 °C. The mixture was poured into ice water (6 mL) and then extracted with DCM (10 mL × 2). The combined organic layers were washed with brine (6 mL), dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM / EtOAc = 10 to 1:1) to give the title compound (11 mg, yield: 6%) as a grayish-white solid. LC-MS (ESI): m / z 320.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ12.35(s,1H),8.06(s,1H),7.84(d,J=8.8Hz,1H),7.65(d,J=8.4Hz ,1H),7.27(d,J=3.6Hz,1H),7.17(d,J=2.0Hz,1H),7.03(dd,J=2.4,8.4Hz,1H),3.85(s,3H).

[0207] Example 2

[0208]

[0209] 2,4-Dimethoxy-N-(5-phenyl-1,3,4-oxadiazol-2-yl)benzamide

[0210] Following the same procedure as in Example 1, using 5-phenyl-1,3,4-oxadiazol-2-amine (186 mg, 1.16 mmol) as the amine, the title compound (45 mg, yield: 12%) was obtained as a white solid. LC-MS (ESI): m / z 326.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ11.13(s,1H),7.98-7.96(m,2H),7.77(d,J=8.8Hz,1H),7.63-7.61(m,3H),6.73-6.69(m,2H),3.93(s,3H),3.87(s,3H).

[0211] Example 3

[0212]

[0213] (S)-1-Benzyl-N-(5-(furan-2-yl)-1,3,4-oxadiazol-2-yl)pyrrolidine-3-carboxamide

[0214] NaH (0.29 g, 7.2 mmol) was added to a solution of 5-(2-furanyl)-1,3,4-oxadiazol-2-amine (0.31 g, 2.0 mmol) in THF (30.0 mL) at 0 °C. The mixture was stirred for 15 min. A solution of (S)-1-benzylpyrrolidine-3-carbonyl chloride (0.8 g, 3.1 mmol) in THF (20 mL) was added at 0 °C. The reaction mixture was stirred at 25 °C for 6 h. The mixture was poured into ice water (20 mL) and then extracted with EtOAc (50 mL × 2). The combined organic layers were washed with brine (40 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC, yielding the title compound as a white solid in two fractions (142 mg, 98.9% purity and 178 mg, 86.6% purity, overall yield: 47%). LC-MS(ESI): m / z 339.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ8.03 (s, 1H), 7.32-7.22 (m, 7H), 6.78 (dd, J = 1.6, 3.2Hz, 1H), 3.59(s,2H),3.35(m,1H),2.83(t,J=8.4Hz,1H),2.62-2.59(m,3H),2.05-1.99(m,2H).

[0215] Example 4

[0216]

[0217] N-(5-(4-hydroxyphenyl)-1,3,4-oxadiazol-2-yl)-2,4-dimethoxybenzamide

[0218] a) 2-(4-(benzooxy)benzoyl)hydrazine-1-thiocarboxamide

[0219] Thioaminourea (554 mg, 6.07 mmol) was added fractionally to a solution of 4-benzyloxybenzoyl chloride (1.0 g, 4.05 mmol) in THF (20 mL) at 15 °C. The reaction mixture was stirred at room temperature for 3 hours. A saturated aqueous solution of NaHCO3 (10 mL) was added, and the resulting precipitate was collected by filtration. The solid was washed with hot H2O (50 °C, 100 mL) and dried to give the title compound (1.1 g, yield: 90%) as a white solid. LC-MS (ESI): m / z 302.2 [M+H] + . 1H NMR (400MHz, DMSO-d6): δ10.25(br s,1H),9.30(br s,1H),7.86(d,J=8.8Hz,3H),7.60(br s,1H),7.47-7.32(m,5H),7.08(d,J=8.8Hz,2H),5.19(s,2H).

[0220] b) 5-(4-(benzooxy)phenyl)-1,3,4-oxadiazol-2-amine

[0221] A solution of KI (182 mg, 1.1 mmol) in H₂O (2 mL) was added to a solution of 2-(4-(benzooxy)benzoyl)hydrazide-1-thiocarboxamide (1.1 g, 3.65 mmol) in i-PrOH (9 mL) at room temperature. The solution was cooled to 5 °C, and an aqueous solution of NaOH (5 M, 1.1 mL) was added. A solution of 1,3-dibromo-5,5-dimethylhydantoin (783 mg, 2.74 mmol) in acetonitrile (11 mL) was added dropwise to the resulting mixture at 5–10 °C. The reaction mixture was stirred at 10 °C for 1 hour. The reaction mixture was quenched with an aqueous solution of NaHSO₃ (0.25 mL). EtOAc (30 mL) was added to the slurry, and the precipitate was collected by filtration. The solid was washed with EtOAc (20 mL) and dried to give the title compound (0.6 g, yield: 60%) as a white solid. LC-MS(ESI): m / z 367.7 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ7.76-7.72(m,2H),7.46-7.35(m,5H),7.17-7.15(m,4H),5.18(s,2H).

[0222] c)N-(5-(4-(benzooxy)phenyl)-1,3,4-oxadiazol-2-yl)-2,4-dimethoxybenzamide

[0223] A solution of 2,4-dimethoxybenzoyl chloride (269 mg, 1.34 mmol) in DCM (10 mL) was added dropwise to a solution of 5-(4-(benzyloxy)phenyl)-1,3,4-oxadiazol-2-amine (300 mg, 1.12 mmol) in pyridine (8 mL) at 10 °C. The reaction mixture was stirred at room temperature for 3 hours. A solution of 2,4-dimethoxybenzoyl chloride (100 mg, 0.50 mmol) in DCM (3 mL) was added, and the reaction mixture was stirred at room temperature overnight. The mixture was poured into ice water (8 mL) and then extracted with DCM (10 mL × 2). The combined organic layers were washed with brine (6 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc / DCM = 3:1:1 to 0:1:1) to give the title compound as a white solid (141 mg, yield: 29%). LC-MS (ESI): m / z 432.3 [M+H] + . 1 H NMR (400MHz, DMSO-d6): δ11.03(s,1H),7.91(d,J=8.8Hz,2H),7.77(d,J=8.8Hz,1H),7.50-7 .40(m,5H),7.24(d,J=8.8Hz,2H),6.72-6.71(m,2H),5.22(s,2H),3.93(s,3H),3.87(s,3H).

[0224] d)N-(5-(4-hydroxyphenyl)-1,3,4-oxadiazol-2-yl)-2,4-dimethoxybenzamide

[0225] Pd / C (10%, 10 mg) was added to a solution of N-(5-(4-(benzoxy)phenyl)-1,3,4-oxadiazol-2-yl)-2,4-dimethoxybenzamide (70 mg, 0.16 mmol) in EtOH (150 mL). The mixture was stirred at room temperature for 12 hours under hydrogen balloon pressure. The catalyst was filtered off, and the filtrate was concentrated under reduced pressure to give the title compound (32 mg, 58%) as a gray solid. LC-MS (ESI): m / z 340.1 [MH] - . 1 H NMR (400MHz, DMSO-d6): δ10.98(s,1H),10.33(s,1H),7.80-7.76(m,3H),6.95(d,J=8.4Hz,2H),6.72-6.68(m,2H),3.93(s,3H),3.86(s,3H).

[0226] Example 5

[0227]

[0228] 2,4-Dimethoxy-N-(5-(thiophen-3-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0229] a) Thiophene-3-carbazide

[0230] A mixture of methyl thiophene-3-carboxylate (0.5 g, 3.52 mmol, 1 eq) and hydrazine hydrate (1.85 g, 35.17 mmol, 1.80 mL, 10 eq) in ethanol (7 mL) was heated to 79 °C for 16 hours. The mixture was cooled to room temperature and concentrated under vacuum to give the title compound (0.46 g, yield: 82%) as a red solid, which was used in the next step. LC-MS (ESI): m / z 143.2 [M+H] + .

[0231] b) 5-(thien-3-yl)-1,3,4-oxadiazol-2-amine

[0232] CH3CN (1.2 mL) containing BrCN (0.4 mL, 2 eq) was added dropwise to a mixture of thiophene-3-carbazide (0.46 g, 2.91 mmol) and K2CO3 (804 mg, 5.82 mmol) in H2O (20 mL) at 25 °C. The mixture was stirred at 25 °C for 16 hours. The mixture was filtered. The solid was washed with water (5 mL) and dried under vacuum to give the title compound (0.41 g, yield: 80%) as a white solid, which was used in the next step. LC-MS (ESI): m / z 168.1 [M+H] + .

[0233] c) 2,4-Dimethoxy-N-(5-(thien-3-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0234] T3P (1.3 mL, 2.20 mmol, 50% in EA, v / v) was added to a mixture of 5-(thien-3-yl)-1,3,4-oxadiazol-2-amine (106 mg, 0.6 mmol), 2,4-dimethoxybenzoic acid (0.1 g, 0.6 mmol), and TEA (0.3 mL, 2.20 mmol) in EtOAc (2 mL), and the mixture was heated to 80 °C for 16 h. The reaction mixture was diluted with water (10 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated under vacuum. The crude product was wet-milled with CH3CN (1 mL) at 25 °C for 30 min, filtered, and dried in a drying oven (30 min) to give the title compound (25 mg, yield: 13%) as a white solid. LC-MS (ESI): m / z 332.1 [M+H] + . 1H NMR (DMSO-d6, 400MHz): δ11.02(s,1H),8.30(dd,J=2.9,1.2Hz,1H),7.83(dd,J=5.0,2.9Hz,1H),7.78(d,J=8. 6Hz,1H),7.61(dd,J=5.1,1.3Hz,1H),6.66-6.77(m,2H),3.94(s,3H),3.87(s,3H),3.83(s,1H),3.81(s,1H).

[0235] The following compounds were synthesized according to the procedure described in Example 5.

[0236]

[0237]

[0238] Example 6

[0239]

[0240] N-(6-(furan-2-yl)pyridazin-3-yl)-2,4-dimethoxybenzamide

[0241] a) N-(6-Chlorpyridazin-3-yl)-2,4-Dimethoxybenzamide

[0242] A solution of 2,4-dimethoxybenzoyl chloride (813 mg, 4.05 mmol) in dichloromethane (5 mL) was added dropwise to a solution of 6-chloropyridazine-3-amine (350 mg, 2.7 mmol) in pyridine (10 mL) at room temperature. The reaction mixture was stirred overnight at room temperature. The mixture was poured into ice water (10 mL) and then extracted with DCM (10 mL × 2). The combined organic layers were washed with brine (10 mL), dried over anhydrous Na₂SO₄, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc = 3:1 to 1:1) to give the title compound (261 mg, yield: 32%).

[0243] b) N-(6-(furan-2-yl)pyridazin-3-yl)-2,4-dimethoxybenzamide

[0244] Under a nitrogen atmosphere, Na₂CO₃ (173 mg, 1.64 mmol) and Pd(PPh₃)₄ (31 mg, 0.3 mmol) were added to a mixture of N-(6-chloropyridazin-3-yl)-2,4-dimethoxybenzamide (401 mg, 1.37 mmol) and furan-2-ylboronic acid (183 mg, 1.64 mmol) in DMF / H₂O (3:2, 10 mL). The reaction mixture was stirred overnight at 90 °C. Water (10 mL) was added, and the resulting mixture was extracted with EtOAc (20 mL × 2). The combined organic layers were washed with brine (20 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether / EtOAc / DCM = 2:1:0.5) to give the title compound (95 mg, yield: 21%) as a white solid. LC-MS(ESI): m / z 326.1 [M+H] + . 1 HNMR (400MHz, DMSO-d6): δ10.85(s,1H),8.55(d,J=9.2Hz,1H),8.08(d,J=9.6Hz,1H), 7.98-7.95(m,2H),7.27(d,J=3.2Hz,1H),6.80-6.74(m,3H),4.07(s,3H),3.88(s,3H).

[0245] Example 7

[0246]

[0247] 4-(benzoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0248] a) Methyl 4-(benzooxy)-2-hydroxybenzoate

[0249] K₂CO₃ (4.94 g, 35.7 mmol) was added to a solution of methyl 2,4-dihydroxybenzoate (2.0 g, 11.9 mmol) in DMF (20 mL) at 5 °C. The mixture was stirred at room temperature for 1 hour, and BnBr (2.43 g, 14.2 mmol) was added dropwise at 5 °C. The reaction mixture was stirred overnight at room temperature. The mixture was diluted with ethyl acetate (20 mL) and washed with brine (50 mL × 3). The combined organic layers were dried over Na₂SO₄, filtered, and evaporated. The residue was purified by silica gel column chromatography (PE / EA = 100:1 to 10:1) to give the title compound (2.3 g, yield: 74%) as a white solid.

[0250] b) Methyl 4-(benzooxy)-2-methoxybenzoate

[0251] NaH (60% in mineral oil, 535 mg, 13.8 mmol) was added fractionally to a solution of methyl 4-(benzooxy)-2-hydroxybenzoate (2.3 g, 8.9 mmol) in anhydrous DMF (10 mL) at 0 °C, and the mixture was stirred at 0 °C for 10 min. Iodimethane (3.16 g, 22.3 mmol) was added dropwise at 0 °C. The reaction mixture was stirred overnight at room temperature. The reaction mixture was quenched with ice water (50 mL) to give a solid. The solid was collected by filtration, washed with water, and dried under vacuum to give the title compound (2.4 g, yield: 97%) as a white solid.

[0252] c) 4-(benzooxy)-2-methoxybenzoic acid

[0253] Anhydrous NaOH (4M, 7 mL, 22 mmol) was added to a solution of methyl 4-(benzoxy)-2-methoxybenzoate (2.4 g, 8.8 mmol) in THF (10 mL) at 0 °C. The reaction mixture was refluxed for 4 hours. The THF was evaporated, and the pH of the mixture was adjusted to 3–4 with dilute aqueous HCl. The mixture was extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with 0.6 N aqueous HCl (20 mL), dried over Na₂SO₄, filtered, and evaporated to give the title compound (2.2 g, yield: 96%) as a white solid.

[0254] d) 2-Methoxy-4-((4-methoxybenzyl)oxy)benzoyl chloride

[0255] At 0 °C and under N2, DMF (2 mL) and oxaloyl chloride (4.0 g, 38 mmol) were added to a solution of 2-methoxy-4-((4-methoxybenzyl)oxy)benzoic acid (491 mg, 1.9 mmol) in DCM (30 mL). The reaction mixture was stirred at room temperature for 30 minutes. The solvent was evaporated to provide the title compound, which was used in the next step without further purification.

[0256] e) 4-(benzooxy)-2-methoxy-N-(5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0257] NaH (191 mg, 4.76 mmol) was added to a solution of 5-(thien-2-yl)-1,3,4-oxadiazol-2-amine (200 mg, 1.19 mmol) in THF (15 mL) at 0 °C under N2. The reaction mixture was stirred at room temperature for 30 min, and 4-(benzyloxy)-2-methoxybenzoyl chloride (509 mg, 1.89 mmol) was added to the mixture at 0 °C. The reaction mixture was stirred at room temperature overnight. The mixture was quenched with water (15 mL) and extracted with ethyl acetate (15 mL × 2). The combined organic layers were washed with brine (40 mL), dried over Na2SO4, filtered, and evaporated. The residue was purified by preparative HPLC to give the title compound (24 mg, yield: 15%) as a white solid. LCMS (ESI): m / z 406.0 [M+H] + . 1 HNMR (300MHz, DMSO-d6): δ7.93(d,J=6.8Hz,1H),7.77-7.74(m,2H),7.50-7.28(m,6H),6.82-6.76(m,2H),5.23(s,2H),3.91(s,3H).

[0258] Example 8

[0259]

[0260] 4-Hydroxy-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0261] a) Methyl 2-hydroxy-4-((4-methoxybenzyl)oxy)benzoate

[0262] K₂CO₃ (9.0 g, 65.5 mmol) was added to a solution of methyl 2,4-dihydroxybenzoate (10.0 g, 59.5 mmol) in DMF (150 mL) at 5 °C. The mixture was stirred at room temperature for 1 hour, and PMBCl (9.32 g, 59.5 mmol) was added dropwise at 5 °C. The reaction mixture was stirred overnight at room temperature. The mixture was diluted with ethyl acetate (200 mL) and washed with brine (25 mL × 3). The organic layer was dried over Na₂SO₄, filtered, and evaporated. The residue was purified by silica gel column chromatography (PE / EA = 100:1 to 10:1) to give the title compound (3.12 g, yield: 16%) as a gray solid. LCMS (ESI-MS): m / z 289.1 [M+H] + .

[0263] b) Methyl 2-methoxy-4-((4-methoxybenzyl)oxy)benzoate

[0264] NaH (625 mg, 15.6 mmol) was added fractionally to a solution of methyl 2-hydroxy-4-((4-methoxybenzyl)oxy)benzoate (3.0 g, 10.40 mmol) in anhydrous DMF (40 mL) at 0 °C, and the mixture was stirred at 0 °C for 10 min. Iodimethane (3.7 g, 26 mmol) was added dropwise at 0 °C, and the reaction mixture was stirred overnight at room temperature. The reaction mixture was quenched with ice water (50 mL), and the resulting solid was collected by filtration. The solid was washed with water and dried under vacuum to give the title compound (2.6 g, yield: 86%) as a white solid. LCMS (ESI-MS): m / z 303.1 [M+H] + .

[0265] c) 2-Methoxy-4-((4-methoxybenzyl)oxy)benzoic acid

[0266] Anhydrous NaOH (4M, 2 mL, 34.4 mmol) was added to a solution of methyl 2-methoxy-4-((4-methoxybenzyl)oxy)benzoate (2.6 g, 8.6 mmol) in THF (10 mL) at 0 °C. The reaction mixture was refluxed for 4 hours. The solvent was evaporated, and the pH of the mixture was adjusted to 5–6 with dilute aqueous HCl. The mixture was extracted with ethyl acetate (50 mL × 3). The combined organic layers were washed with 0.6 N aqueous HCl (100 mL), dried over Na₂SO₄, filtered, and evaporated to give the title compound (1.6 g, yield: 65%) as a white solid. LCMS (ESI-MS): m / z 289.1 [M+H] + .

[0267] d) 2-Methoxy-4-((4-methoxybenzyl)oxy)benzoyl chloride

[0268] DMF (2 μL) and oxaloyl chloride (1.7 g, 13.8 mmol) were added to a solution of 2-methoxy-4-((4-methoxybenzyl)oxy)benzoic acid (200 mg, 1.03 mmol) in dichloromethane (20 mL) at 0 °C under N2. The reaction mixture was stirred at room temperature for 30 minutes. The solvent was evaporated to provide the title compound, which was used in the next step without further purification.

[0269] e)2-Methoxy-4-((4-methoxybenzyl)oxy)-N-(5-(thien-2-yl)-1,3,4-oxadiazole-2- (Bento) benzamide

[0270] NaH (60% in mineral oil, 110 mg, 2.76 mmol) was added to a solution of 5-(thiophene-2-yl)-1,3,4-oxadiazol-2-amine (116 mg, 0.69 mmol) in THF (5 mL) at 0 °C under N2. The reaction mixture was stirred at room temperature for 30 min, and 2-methoxy-4-((4-methoxybenzyl)oxy)benzoyl chloride (211 mg, 0.69 mmol) was added to the mixture at 0 °C. The reaction mixture was stirred overnight at room temperature. The mixture was quenched with water (10 mL) and extracted with ethyl acetate (10 mL × 2). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and evaporated. The residue was purified by preparative HPLC to give the title compound (60 mg, yield: 20%) as a white solid. LCMS (ESI-MS): m / z 438.1 [M+H] + .

[0271] f) 4-Hydroxy-2-methoxy-N-(5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0272] Trifluoroacetic acid (2 mL) was added to a solution of 2-methoxy-4-((4-methoxybenzyl)oxy)-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (60 mg, 0.14 mmol) in 2 mL of DCM at room temperature. The reaction mixture was stirred at room temperature for 1 hour. The DCM was evaporated under reduced pressure to give the title compound (18 mg, yield: 42%) as a white solid. LCMS (ESI-MS): m / z 318.1 [M+H] + . 1 HNMR (300MHz, DMSO-d6): δ10.89(s,1H),10.41(s,1H),7.94-7.92(m,1H),7.77-7.69(m,2H),7.31-7.26(m,1H),6.58-6.49(m,2H),3.89(s,3H).

[0273] Example 9

[0274]

[0275] 4-(2-hydroxyethoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0276] a) Methyl 4-(2-(benzooxy)ethoxy)-2-hydroxybenzoate

[0277] K₂CO₃ (4.3 g, 30 mmol) was added to a solution of methyl 2,4-dihydroxybenzoate (5.0 g, 30 mmol) in DMF (100 mL) at 5 °C. The mixture was stirred at room temperature for 1 hour, and ((2-bromoethoxy)methyl)benzene (6.4 g, 30 mmol) was added dropwise at 5 °C. The reaction mixture was stirred overnight at room temperature. The mixture was diluted with ethyl acetate (60 mL) and then washed with brine (200 mL × 3). The combined organic layers were dried over Na₂SO₄, filtered, and evaporated. The residue was purified by silica gel column chromatography (PE / EA = 100:1 to 10:1) to give the title compound (2.16 g, yield: 24%) as a white solid.

[0278] b) Methyl 4-(2-(benzooxy)ethoxy)-2-methoxybenzoate

[0279] Pd / C (5%, 180 mg, 0.57 mmol) was added to a solution of methyl 4-(2-(benzoxy)ethoxy)-2-hydroxybenzoate (1.8 g, 5.7 mmol) in MeOH (5 mL) at room temperature. The reaction mixture was stirred at room temperature under hydrogen balloon pressure for 12 hours. The reaction mixture was filtered and the filtrate was collected. The solvent in the filtrate was evaporated to give the title compound (900 mg, yield: 70%) as a white solid, which was used in the next step without further purification.

[0280] c) Methyl 4-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-2-methoxybenzoate

[0281] Imidazole (320 mg, 4.7 mmol) was added to a solution of methyl 4-(2-(benzyloxy)ethoxy)-2-methoxybenzoate (900 mg, 3.96 mmol) in anhydrous THF (30 mL) at 0 °C. TBSCl (661 mg, 4.38 mmol) was added, and the mixture was stirred overnight at room temperature. The reaction mixture was quenched with water (10 mL) and extracted with EA (10 mL × 2). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄, filtered, and concentrated under vacuum to give the title compound (680 mg, yield: 50%) as a white solid. LCMS (ESI-MS): m / z 327.1 [M+H] + .

[0282] d) 4-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-2-methoxybenzoic acid

[0283] Anhydrous NaOH (4M, 2mL, 8mmol) was added to a solution of methyl 4-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-2-methoxybenzoate (680 mg, 2 mmol) in THF (10 mL) at 0 °C. The reaction mixture was refluxed for 4 hours. The solvent was evaporated, and the pH of the mixture was adjusted to 5–6 with dilute aqueous HCl. The mixture was extracted with ethyl acetate (10 mL × 3). The combined organic layers were washed with 0.6N aqueous HCl (20 mL), dried over Na₂SO₄, filtered, and evaporated to give the title compound (450 mg, yield: 66%) as a white solid, which was used in the next step without further purification.

[0284] e)4-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-2-methoxybenzoyl chloride

[0285] DMF (2 mL) and oxaloyl chloride (2.73 g, 21.5 mmol) were added to a solution of 4-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-2-methoxybenzoic acid (350 mg, 1.07 mmol) in DCM (10 mL) at 0 °C under N2. The reaction mixture was stirred at room temperature for 30 minutes. The solvent was evaporated under reduced pressure to provide the title compound, which was used in the next step without further purification.

[0286] f)4-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1, 3,4-Oxadiazol-2-yl)benzamide

[0287] NaH (60% in mineral oil, 70 mg, 2.72 mmol) was added to a solution of 5-(thiophen-2-yl)-1,3,4-oxadiazol-2-amine (112 mg, 0.68 mmol) in THF (5 mL) at 0 °C under N2. The reaction mixture was stirred at room temperature for 30 min, and 4-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-2-methoxybenzoyl chloride (370 mg, 1.07 mmol) was added to the mixture at 0 °C. The reaction mixture was stirred at room temperature overnight. The mixture was quenched with water (5 mL) and extracted with ethyl acetate (10 mL × 2). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and evaporated. The residue was purified by preparative HPLC to give the title compound (20 mg, yield: 17%) as a white solid.

[0288] g)4-(2-hydroxyethoxy)-2-methoxy-N-(5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)benzoyl amine

[0289] TFA (2 mL) was added to a solution of 4-(2-((tert-butyldimethylsilyl)oxy)ethoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (20 mg, 0.04 mmol) in DCM (2 mL) for 12 hours at room temperature. The reactants were filtered, and the solvent in the filtrate was evaporated under reduced pressure to give the title compound (7 mg, yield: 46%) as a white solid. LCMS (ESI-MS): m / z 360.1 [M+H] + . 1 HNMR (400MHz, DMSO-d6): δ11.06(s,1H),7.95(s,1H),7.78-7.75(m,2H),7.31-7.29(m,1H),6. 72-6.69(m,2H),4.92(t,J=5.6Hz,1H),4.12(t,J=4.8Hz,2H),3.93(s,3H),3.77-3.74(m,2H).

[0290] Example 10

[0291]

[0292] 4-(2-(benzyloxy)ethoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzene formamide

[0293] a) Methyl 4-(2-(benzooxy)ethoxy)-2-hydroxybenzoate

[0294] K₂CO₃ (4.3 g, 3 mmol) was added to a solution of methyl 2,4-dihydroxybenzoate (5.0 g, 30 mmol) in DMF (100 mL) at 5 °C. The mixture was stirred at room temperature for 1 hour, and ((2-bromoethoxy)methyl)benzene (6.4 g, 30 mmol) was added dropwise at 5 °C. The reaction mixture was stirred overnight at room temperature. The mixture was diluted with ethyl acetate (60 mL), washed with brine (200 mL × 3), and the combined organic layers were dried over Na₂SO₄, filtered, and the solvent was evaporated. The residue was purified by silica gel column chromatography (PE / EA = 100:1 to 10:1) to give the title compound (2.16 g, yield: 24%) as a white solid.

[0295] b) 4-(2-(benzooxy)ethoxy)-2-methoxybenzoic acid

[0296] Anhydrous NaOH (5 mL, 1.14 g, 28.5 mmol) was added to a solution of methyl 4-(2-(benzyloxy)ethoxy)-2-methoxybenzoate (2.16 g, 7.1 mmol) in 50 mL of THF at 0 °C. The reaction mixture was refluxed for 4 hours. The THF was evaporated under reduced pressure, and the pH of the mixture was adjusted to 5–6 with dilute HCl. The mixture was extracted with EA (50 mL × 3). The combined organic layers were washed with an aqueous HCl solution (0.6 N, 100 mL), dried over Na₂SO₄, filtered, and evaporated to give the title compound (1.8 g, yield: 63%) as a white solid. LCMS (ESI-MS): m / z 303.1 [M+H] + .

[0297] c) 4-(2-(benzooxy)ethoxy)-2-methoxybenzoyl chloride

[0298] At 0 °C and under N2, DMF (2 mL) and (COCl)2 (2.5 g, 20.7 mmol) were added to a solution of 4-(2-(benzoxy)ethoxy)-2-methoxybenzoic acid (300 mg, 1.03 mmol) in DCM (20 mL). The reaction mixture was stirred at room temperature for 30 minutes. The DCM was evaporated to provide the title compound, which was used in the next step without further purification.

[0299] d) 4-(2-(benzyloxy)ethoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl) benzamide

[0300] NaH (60% in mineral oil, 166 mg, 4.16 mmol) was added to a solution of 5-(thiophene-2-yl)-1,3,4-oxadiazol-2-amine (534 mg, 1.67 mmol) in THF (20 mL) at 0 °C under N2. The reaction mixture was stirred at room temperature for 30 min, and 4-(2-(benzyloxy)ethoxy)-2-methoxybenzoyl chloride (280 mg, 1.67 mmol) was added to the mixture at 0 °C. After the addition, the reaction mixture was stirred at room temperature overnight. The mixture was quenched with water (10 mL) and extracted with EA (20 mL × 2). The combined organic layers were washed with brine (40 mL), dried over Na2SO4, filtered, and evaporated. The residue was purified by preparative HPLC to give the title compound (40 mg, yield: 5%) as a white solid. LCMS (ESI-MS): m / z 452.1 [M+H] + . 1HNMR (300MHz, DMSO-d6): δ11.08 (s, 1H), 7.93 (d, J = 4.5Hz, 1H), 7.78-7.75 (m, 2H), 7.37-7.2 7(m,6H),6.74-6.68(m,2H),4.58(s,2H),4.45-4.39(m,2H),3.92(s,3H),3.82-3.76(m,2H).

[0301] Example 11

[0302]

[0303] 2-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenoxy)ethyl acid

[0304] a) 2-Methoxy-4-(2-oxoethoxy)-N-(5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)benzoyl amine

[0305] To a solution of 4-(2,3-dihydroxypropoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (47 mg, 0.12 mmol) in dichloromethane (2 mL), NaIO4 (34 mg, 0.16 mmol) was added, and the mixture was stirred overnight at room temperature. The reaction mixture was extracted with ethyl acetate (30 mL × 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under vacuum to give the title compound (60 mg, 100%) as a white solid, which was used in the next step without further purification.

[0306] b) 2-(3-methoxy-4-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenoxy) Acetic acid

[0307] To a solution of 2-methoxy-4-(2-oxoethoxy)-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (60 mg, 0.17 mmol) in acetone (2 mL), NaClO2 (104 mg, 0.92 mmol) and NaH2PO4 (61 mg, 0.50 mmol) were added. The mixture was stirred at room temperature for 12 hours. The reaction mixture was extracted with ethyl acetate (30 mL × 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by preparative HPLC to give the title compound (4 mg, yield: 6%) as a white solid. LCMS (ESI-MS): m / z 376.0 [M+H] + . 1HNMR (400MHz, DMSO-d6): δ13.10(s,br,1H),11.40(s,1H),7.93(d,J=8Hz,1H),7.60(d,J=8Hz,1H),7 .20(d,J=8Hz,1H),7.29(d,J=8.0Hz,1H),6.74(s,1H),6.70-.6.64(m,1H),4.80(s,2H),3.91(s,3H).

[0308] Example 12

[0309]

[0310] 4-(2-(dimethylamino)ethoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl) benzamide

[0311] Dimethylamine (0.4 mL, 0.70 mmol) and acetic acid (0.1 mL) were added to a solution of 2-methoxy-4-(2-oxoethoxy)-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (50 mg, 0.14 mmol) in DCM (2 mL). The mixture was allowed to react at room temperature for 1 hour. NaBH(OAc)3 (45 mg, 0.21 mmol) was added, and the mixture was stirred overnight at room temperature. The reaction mixture was extracted with ethyl acetate (30 mL × 3). The combined organic layers were washed with brine, dried over Na2SO4, and concentrated under vacuum. The residue was purified by preparative HPLC to give the title compound (4 mg, yield: 7%) as a white solid. LCMS (ESI-MS): m / z 388.9 [M+H] + . 1 H NMR (400MHz, CD3OD): δ8.04(d,J=8.4Hz,1H),7.85-7.79(m,2H),7.28(t,J=4.8Hz,1H),6 .83-6.81(m,2H),4.51(t,J=4.4Hz,1H),4.10(s,3H),3.68(t,J=4.4Hz,2H),3.05(s,6H).

[0312] Example 13

[0313]

[0314] 2-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenoxy)-2- Methylpropionic acid

[0315] a) Ethyl 2-(4-formyl-3-methoxyphenoxy)-2-methylpropionate

[0316] Cs₂CO₃ (321.22 mg, 0.99 mmol) was added to a suspension of 4-hydroxy-2-methoxybenzaldehyde (0.04 mL, 0.3 mmol) and ethyl 2-bromo-2-methylpropionate (0.14 mL, 0.99 mmol) in DMF (1 mL) at 25 °C. The reaction mixture was stirred at 50 °C for 16 hours. The mixture was diluted with H₂O (5 mL) and extracted with ethyl acetate (5 mL × 3). The combined organic layers were dried over MgSO₄ and filtered. The filtrate was concentrated under vacuum. The residue was dissolved in DCM (1 mL) and washed with NaOH (1 M, 1 mL × 3). The combined organic layers were separated and the solvent was removed under vacuum to give the title compound (50 mg, 51%) as a yellow oil. LC-MS (ESI): m / z 267.0 [M + H] + .

[0317] b) 4-((1-ethoxy-2-methyl-1-oxopropyl-2-yl)oxy)-2-methoxybenzoic acid

[0318] A solution of ethyl 2-(4-formyl-3-methoxyphenoxy)-2-methylpropionate (50 mg, 0.17 mmol) in t-BuOH (1.50 mL) was added to H₂O (0.67 mL) containing NaClO₂ (141 mg, 1.6 mmol) and NaH₂PO₄ (120 mg, 1.1 mmol). The mixture was stirred at 25 °C for 16 hours. The mixture was a yellow solution. The mixture was diluted with H₂O (5 mL) and extracted with EtOAc (5 mL × 3). The combined organic layers were dried over MgSO₄, filtered, and the filtrate was concentrated under vacuum to give the title compound (35 mg, yield: 66%) as a yellow solid. LC-MS (ESI): m / z 238.2 [M+H] + .

[0319] c) 2-(3-methoxy-4-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenoxy)- ethyl 2-methylpropionate

[0320] T3P (266 mL, 0.45 mmol) was added to a suspension of 4-((1-ethoxy-2-methyl-1-oxopropyl-2-yl)oxy)-2-methoxybenzoic acid (35 mg, 0.11 mmol), 5-(thiophen-2-yl)-1,3,4-oxadiazol-2-amine (28 mg, 0.17 mmol), and TEA (60 mL, 0.45 mmol) in ethyl acetate (1 mL) at 25 °C, and the reaction mixture was stirred at 80 °C for 16 hours. The mixture was diluted with H2O (1 mL) and extracted with ethyl acetate (2 mL × 3). The combined organic layers were dried over MgSO4 and filtered. The filtrate was concentrated under vacuum to give the title compound (65 mg, yield: 68%) as a yellow oil. LC-MS (ESI): m / z 432.2 [M+H] + .

[0321] d)2-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenoxy)- 2-Methylpropionic acid

[0322] LiOH (7.33 μL, 0.26 mmol) was added to a solution of ethyl 2-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenoxy)-2-methylpropionate (65 mg, 0.075 mmol) in THF (0.5 mL) and H₂O (0.5 mL) at 25 °C. The mixture was stirred at 50 °C for 16 h. The solvent was removed under vacuum. The residue was dissolved in H₂O (3 mL) and the pH of the mixture was adjusted to 4 using 1 N HCl aqueous solution. The mixture was purified by preparative HPLC (column: Phenomenex Gemini C18 250*50 mm*10 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 38%-58%, 10 min) to give the title compound (5 mg, yield: 16%) as a white solid. LC-MS(ESI): m / z 404.1 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ11.17(br s,1H),7.94(d,J=4.0Hz,1H),7.76(d,J=2.7Hz,1H),7.69(d,J=8.7Hz,1H),7.29(dd,J=3.8 ,4.9Hz,1H),6.61(d,J=2.1Hz,1H),6.48(dd,J=2.2,8.7Hz,1H),3.87(s,3H),1.59(s,6H).

[0323] Example 14

[0324]

[0325] 4-((2H-tetrazol-5-yl)methoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl) benzamide

[0326] a) Methyl 4-(cyanomethoxy)-2-hydroxybenzoate

[0327] 2-Bromoacetonitrile (3.57 g, 30 mmol) was added to a solution of methyl 2,4-dihydroxybenzoate (5.0 g, 30 mmol) and K₂CO₃ (4.32 g, 31 mmol) in DMF (50 mL) at 0 °C. The mixture was stirred at 25 °C for 12 hours under a nitrogen atmosphere. The mixture was filtered, and the solvent was concentrated under vacuum. The solvent was then analyzed by rapid silica gel chromatography (…). 80g The residue was purified by rapid silica gel column chromatography (eluent: 0–50% ethyl acetate / petroleum ether gradient, at 30 mL / min) to give the title compound (1.6 g, yield: 26%) as a white solid. LC-MS (ESI): m / z 207.9 [M+H] + .

[0328] b) Methyl 4-(cyanomethoxy)-2-methoxybenzoate

[0329] NaH (60% in mineral oil, 0.25 g, 6.3 mmol) was added to a solution of methyl 4-(cyanomethoxy)-2-hydroxybenzoate (1.0 g, 4.8 mmol) in DMF (20 mL) at 0 °C. The solution was stirred at 25 °C for 0.5 h, and iodomethane (0.89 g, 6.3 mmol) was added dropwise at 0 °C. The reaction mixture was stirred at 25 °C under N2 for 12 h. Water (60 mL) was added, and the mixture was extracted with EtOAc (60 mL × 3). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and concentrated to give the title compound (1.05 g, 98%) as a brown oil. LC-MS (ESI): m / z 222.1 [M+H] + .

[0330] c) Methyl 4-((2H-tetrazol-5-yl)methoxy)-2-methoxybenzoate

[0331] Dibutylstannanone (225 mg, 0.90 mmol) was added to a solution of methyl 4-(cyanomethoxy)-2-methoxybenzoate (1.0 g, 4.5 mmol) and trimethyl azidosilane (1.0 g, 9.0 mmol) in toluene (20 mL) at 0 °C. The mixture was stirred at 120 °C under N2 for 12 hours. The mixture was concentrated to approximately 3 mL, and PE (12 mL) and MTBE (3 mL) were added. The mixture was stirred at 25 °C for 3 hours to give a white suspension. The solid was filtered to give the title compound (1.0 g, 84%) as a white solid. LC-MS (ESI): m / z 264.9 [M+H] + .

[0332] d) 4-((2H-tetrazol-5-yl)methoxy)-2-methoxybenzoic acid

[0333] 1 M anhydrous LiOH (0.8 mL, 0.8 mmol) was added to a solution of methyl 4-((2H-tetrazol-5-yl)methoxy)-2-methoxybenzoate (100 mg, 0.38 mmol) in THF (1 mL). The reaction mixture was stirred at 50 °C under N2 for 12 hours. The mixture was concentrated, and the pH of the residue was adjusted to 3–4 with 1 M HCl aqueous solution to give a solid. The solid was filtered to give the title compound (70 mg, 73.9%) as a white solid. LC-MS (ESI): m / z 251.1 [M+H] + .

[0334] e)4-((2H-tetrazol-5-yl)methoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2- (Bento) benzamide

[0335] T3P (50% in EA, 1.0 mL, 1.6 mmol) was added to a solution of 4-((2H-tetrazol-5-yl)methoxy)-2-methoxybenzoic acid (50 mg, 0.20 mmol), 5-(thiophen-2-yl)-1,3,4-oxadiazol-2-amine (67 mg, 0.40 mmol), and TEA (0.11 mL, 0.8 mmol) in ethyl acetate (5 mL). The mixture was stirred at 80 °C under N2 for 12 hours. Water (20 mL) was added, and the mixture was extracted with EtOAc (30 mL × 3). The combined organic layers were dried over Na2SO4, filtered, and concentrated. The residue was purified by preparative HPLC (column: Boston Uni C18 40*150*5um; mobile phase: water (0.225% FA)-ACN; B%: 18%-58%, 10 min) to give the title compound (6 mg, yield: 7.5%) as a yellow solid. LC-MS (ESI): m / z 400.0 [M+H] + .1 H NMR (400MHz, DMSO-d6) δ7.99-7.91(m,1H),7.76(d,J=3.5Hz,1H),7.38(d,J=3 .1Hz,1H),7.33-7.26(m,2H),7.17(d,J=9.0Hz,1H),5.45(s,2H),3.85(s,3H).

[0336] Example 15

[0337]

[0338] 2-Methoxy-4-((2-((5-methyl-2-oxo-1,3-dioxacyclopenten-4-yl)methyl)-2H-tetrazole-5- (5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0339]

[0340] 2-Methoxy-4-((1-((5-methyl-2-oxo-1,3-dioxacyclopenten-4-yl)methyl)-1H-tetrazole-5- (5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0341] Add K₂CO₃ (8 mg, 0.06 mmol) and KI (5 mg, 0.03 mmol) to a solution of 4-((2H-tetrazol-5-yl)methoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (20 mg, 0.05 mmol) and 4-(chloromethyl)-5-methyl-1,3-dioxacyclopenten-2-one (5 μL, 0.045 mmol) in acetonitrile (1 mL). Stir the mixture at 50 °C under N₂ for 1.5 hours. Add a second portion of acetonitrile (1 mL) containing 4-(chloromethyl)-5-methyl-1,3-dioxacyclopenten-2-one (5 μL, 0.045 mmol), and stir the reaction mixture again at 50 °C for 1.5 hours. A third portion of acetonitrile (1 mL) containing 5 μL (0.045 mmol) of 4-(chloromethyl)-5-methyl-1,3-dioxane-2-one was added, and the reaction mixture was stirred at 50 °C for 16 hours. The mixture was diluted with acetonitrile (10 mL) and filtered. The solvent was removed under N2. The residue was purified by preparative HPLC (column: YMC-Actus Triart C18 150*30mm*7um; mobile phase: [water (0.225% FA)-ACN]; B%: 38%-58%, 10 min) to give two fractions of the product as two isomers. Isomer 1 (HPLC: RT = 3.97 min, 4 mg, yield: 15%), LC-MS (ESI): m / z 512.2 [M+H] + ; 1¹H NMR (400MHz, DMSO-d⁶) δ 7.93 (d, J = 5.1Hz, 1H), 7.73 (br d, J = 3.1Hz, 1H), 7.40 (d, J = 2.6Hz, 1H), 7.33–7.23 (m, 2H), 7.21–7.13 (m, 1H), 5.80 (s, 2H), 5.57 (s, 2H), 3.84 (s, 3H), 2.18 (s, 3H); and isomer 2 (HPLC: RT = 4.08 min, 3 mg, yield 12%), as a white solid, LC-MS (ESI): m / z 512.2 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ7.92 (br d, J=4.9Hz, 1H), 7.73 (br s, 1H), 7.34 (br s,1H),7.31-7.20(m,2H),7.17-7.11(m,1H),6.03(s,2H),5.41(s,2H),3.83(s,3H),2.27(s,3H).

[0342] Example 16

[0343]

[0344] 4-(3-cyanopropyl)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0345] a) 5-(thien-2-yl)-1,3,4-oxadiazol-2-amine

[0346] Acetonitrile (20 mL) containing BrCN (9.78 mL, 133 mmol) was added dropwise to a mixture of thiophene-2-carbonylhydrazine (10.5 g, 66.5 mmol) and H₂O (500 mL) at 25 °C. The mixture was stirred at 25 °C for 16 hours. The mixture was filtered, and the white solid was washed with water to give the title compound (4.8 g, yield: 30%) as a yellow solid. LC-MS (ESI): m / z 168.0 [M+H] + .

[0347] b) 4-Bromo-2-methoxy-N-(5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0348] T3P (3.38 mL, 11.4 mmol) was added to a suspension of 5-(thien-2-yl)-1,3,4-oxadiazol-2-amine (500 mg, 2.84 mmol), 4-bromo-2-methoxybenzoic acid (984 mg, 4.26 mmol), and trimethylamine (1.58 mL, 11.4 mmol) in ethyl acetate (20 mL) at 25 °C, and the reaction mixture was stirred at 80 °C for 16 h. The mixture was diluted with H2O (10 mL) and extracted with ethyl acetate (10 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was washed with ethyl acetate (3 mL), and the solid was collected to give the title compound (730 mg, yield: 64%) as a yellow solid. LC-MS (ESI): m / z 380.0 [M+H] + .

[0349] c) 4-(3-cyanopropyl)-2-methoxy-N-(5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0350] Pd(PPh3)4 (30 mg, 0.03 mmol) was added to a mixture of 4-bromo-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (200 mg, 0.5 mmol) and (3-cyanopropyl)zinc bromide(II) (1 mL, 0.5 M solution in THF, 1.05 mmol) in THF (4 mL). The mixture was stirred in a microwave at 100 °C for 30 min. The mixture was filtered through silica and the solvent was removed under vacuum. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX 150*30 mm*5 μm; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 30%-70%, 10 min) to give a solid. The solid was washed with ethyl acetate (3 mL) and collected to give the title compound (21 mg, yield: 11%) as a white solid. LC-MS (ESI): m / z 369.0 [M+H] + . 1 H NMR(DMSO-d6,400MHz): δ11.40(br s,1H),7.93(d,J=4.9Hz,1H),7.74(d,J=3.0Hz,1H),7.63(d,J=8.0Hz,1H),7.28(t,J=4.3Hz,1H),7.08(s,1H),6.95(br d,J=8.3Hz,1H),3.90(s,3H),2.74(br t,J=7.7Hz,2H),2.54(br s,2H),1.84-1.99(m,2H).

[0351] Example 17

[0352]

[0353] 4-(3-(2H-tetrazol-5-yl)propyl)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl) benzamide

[0354] TMSN3 (50 mg, 0.4 mmol) was added to a solution of 4-(3-cyanopropyl)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (200 mg, 0.2 mmol) and Bu₂SnO (7.21 mL, 43 mmol) in toluene (2 mL) at 25 °C. The mixture was stirred at 120 °C for 16 hours. The solvent was removed from the mixture under vacuum, and it was dissolved in DMSO (2 mL) and purified by preparative HPLC (column: Phenomenex Gemini-NX 150*30 mm*5 μm; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 30%-70%, 10 min) to give the title compound (4.6 mg, 5.1%) as a white solid. LC-MS (ESI): m / z 412.1 [M+H] + . 1 H NMR(DMSO-d6,400MHz): δ7.93(d,J=5.0Hz,1H),7.75(d,J=3.3Hz,1H),7.64(d,J=8.0Hz,1H),7.24-7.34(m,1H), 7.07(s,1H),6.96(d,J=8.0Hz,1H),3.90(s,3H),2.90(br,t,J=7.5Hz,2H),2.64-2.76(m,2H),2.01-2.12(m,2H).

[0355] Example 18

[0356]

[0357] cis-4-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenyl) ring Hexane-1-carboxylic acid

[0358]

[0359] trans-4-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenyl) ring Hexane-1-carboxylic acid

[0360] a) 4'-(ethoxycarbonyl)-3-methoxy-2',3',4',5'-tetrahydro-[1,1'-biphenyl]-4-carboxylic acid

[0361] Pd(dppf)Cl2·CH2Cl2 (530 mg, 0.65 mmol) was added to a suspension of 4-bromo-2-methoxybenzoic acid (300 mg, 1.3 mmol), ethyl 4-(4,4,5,5-tetramethyl-1,3,2-dioxane-2-yl)cyclohex-3-en-1-carboxylic acid (437 mg, 1.56 mmol), and K2CO3 (0.40 mL, 3.9 mmol) in dioxane (13 mL) and H2O (1.3 mL) at 25 °C under N2. The compound was stirred at 105 °C for 16 hours. The mixture was filtered, and the filter cake was washed with ethyl acetate (5 mL × 2). The filtrate was extracted with ethyl acetate (5 mL × 3), and the combined organic layers were dried over MgSO4, filtered, and concentrated under vacuum. The residue was washed with ethyl acetate (3 mL) and filtered to give a solid, the title compound (200 mg, yield: 38%), as a brown solid. LC-MS (ESI): m / z 305.2 [M+H] + .

[0362] b) 4-(4-(ethoxycarbonyl)cyclohexyl)-2-methoxybenzoic acid

[0363] Triethylsilane (74 mL, 0.46 mmol), trifluoroacetic acid (17 mL, 0.23 mmol), and Pd(OAc)₂ (15 mg, 0.07 mmol) were added to a suspension of 4'-(ethoxycarbonyl)-3-methoxy-2',3',4',5'-tetrahydro-[1,1'-biphenyl]-4-carboxylic acid (20 mg, 0.05 mmol) in DCM (3 mL) at 25 °C, and the mixture was stirred at 25 °C for 16 hours. The mixture was filtered and washed with DCM (2 mL). The solvent was removed under vacuum to give the title compound (26 mg, yield: 92%) as a yellow oil. LC-MS (ESI): m / z 307.2 [M+H] + .

[0364] c) 4-(3-methoxy-4-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenyl)cyclo Ethyl hexane-1-carboxylate

[0365] T3P (285 mL, 0.5 mmol) was added to a suspension of 5-(thiophen-2-yl)-1,3,4-oxadiazol-2-amine (20 mg, 0.12 mmol), 4-(4-(ethoxycarbonyl)cyclohexyl)-2-methoxybenzoic acid (55 mg, 0.14 mmol), and triethylamine (66 mL, 0.5 mmol) in ethyl acetate (1 mL) at 25 °C. The reaction mixture was stirred at 80 °C for 16 hours. The mixture was diluted with H2O (3 mL) and extracted with ethyl acetate (3 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under vacuum to give the title compound (80 mg, yield: 88%) as a yellow oil. LC-MS (ESI): m / z 456.2 [M+H] + .

[0366] d) 4-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenyl) ring Hexane-1-carboxylic acid

[0367] LiOH (1 mg, 0.367 mmol) was added to a solution of ethyl 4-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenyl)cyclohexane-1-carboxylate (80 mg, 0.11 mmol) in THF (0.5 mL) and H₂O (0.5 mL) at 25 °C. The mixture was stirred at 50 °C for 16 hours. The solvent was removed under vacuum, and the residue was dissolved in H₂O (3 mL). The pH of the mixture was adjusted to 4–5 by adding 1N HCl aqueous solution, and the mixture was purified by preparative HPLC (column: Phenomenex Gemini-NX 150*30mm*5um; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 0%–40%, 10 min) to obtain the title compound as a white solid, which is a pair of separate positional isomers: isomer 1 as a white solid (HPLC: RT = 4.06 min, 5.8 mg, yield: 11%); LC-MS (ESI): m / z 428.2 [M+H) + ; 1¹H NMR (400MHz, DMSO-d6) δ=7.84(br s, 1H), 7.70(br s, 1H), 7.62(s, 1H), 7.26(t, J=4.4Hz, 1H), 6.95(s, 2H), 3.87(s, 3H), 2.10–2.10(m, 1H), 2.07(br s, 1H), 1.91(s, 1H), 1.63–1.42(m, 6H), 1.27(s, 2H); and isomer 2 (HPLC: RT=4.16min, 5mg, yield: 10%): LC-MS (ESI): m / z 428.2 [M+H] + ; 1 H NMR (400MHz, DMSO-d6) δ7.85(s,1H),7.71(s,1H),7.64(s,1H),7.26(d,J=5.0Hz,1H),6.94(s,2H),3.90(br s,3H),2.15(br s,2H),1.79-1.63(m,6H),1.27(br s,2H).

[0368] Example 19

[0369]

[0370] 3'-Methoxy-4'-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)-2,3,4,5-tetracycline Hydro-[1,1'-biphenyl]-4-carboxylic acid

[0371] LiOH (5.18 mg, 0.12 mmol) was added to a solution of ethyl 3'-methoxy-4'-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)-2,3,4,5-tetrahydro-[1,1'-biphenyl]-4-carboxylate (20 mg, 0.04 mmol) in H2O (0.25 mL) and THF (0.25 mL) at 25 °C. The mixture was stirred at 50 °C for 16 hours. The solvent was evaporated under vacuum, and the residue was dissolved in H2O (3 mL). The pH of the mixture was adjusted to pH 4, and the mixture was purified by preparative HPLC (column: Diamonsil C18 150*30mm*5um; mobile phase: [water (0.225% FA)-ACN]; B%: 38%-78%, 10 min) to obtain the title compound (3.2 mg, yield: 21%) as a white solid. LC-MS (ESI): m / z 426.0 [M+H] + . 1H NMR (DMSO-d6, 400MHz): δ7.94 (d, J = 3.9 Hz, 1H), 7.76 (d, J = 2.6 Hz, 1H), 7.67 (d, J = 7.9 Hz, 1H), 7. 27-7.33(m,1H),7.12-7.19(m,2H),6.33-6.43(m,1H),3.94(s,3H),2.56-2.65(m,1H),2.46(br s,2H),2.36-2.42(m,1H),2.11(br d,J=12.0Hz,1H),1.73(br s,1H).

[0372] Example 20

[0373]

[0374] (3-Methoxy-4-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenyl)proline

[0375] CuI (1 mg, 0.005 mmol) and Cs₂CO₃ (34 mg, 0.1 mmol) were added to a suspension of 4-bromo-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (20 mg, 0.053 mmol) and proline (0.02 mL, 0.16 mmol) in DMSO (0.5 mL) under N₂. The mixture was stirred at 120 °C for 16 hours. The mixture was diluted with H₂O (2 mL) and extracted with EtOAc (3 mL × 3). The combined organic layers were dried over MgSO₄ and concentrated under vacuum. The residue was purified by preparative HPLC (column: YMC-Actus Triart C18 150*30mm*7um; mobile phase: [water (0.225% FA)-ACN]; B%: 35%-55%, 10 min) to give the title compound (3.6 mg, yield: 16%) as a yellow solid. LC-MS (ESI): m / z 415.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.37(br s,3H),8.02-7.79(m,1H),7.79-7.59(m,2H),7.28(br s,1H),6.43-5.93(m,2H),3.93(br s,3H),3.75(br s,1H),2.21-1.86(m,6H).

[0376] Example 21

[0377]

[0378] 1-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenyl)piperidine- 3-Formic acid

[0379] RuPhos-Pd-G3 (176 mg, 0.21 mmol) was added to a solution of 4-bromo-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (80 mg, 0.21 mmol), methyl piperidine-3-carboxylate (45 mg, 0.32 mmol), and Cs₂CO₃ (206 mg, 0.63 mmol) in tert-amyl alcohol (2.5 mL) at 25 °C under N₂. The mixture was stirred at 120 °C for 16 h. The solvent was removed under vacuum, and the residue was purified by preparative HPLC (column: Phenomenex Gemini-NX150*30 mm*5 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 26%-66%, 10 min) to give the title compound (6 mg, yield: 6%) as a gray solid. LC-MS(ESI): m / z 429.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ7.94(d,J=5.0Hz,1H),7.68-7.82(m,2H),7.30(t,J=4.3Hz,1H),6.65(br d,J=8.5Hz,1H),6.56(s,1H),3.96(s,3H),3.91(br d,J=12.4Hz,1H),3.75(br d,J=14.0Hz,1H),3.00-3.24(m,3H),1.94(br s,1H),1.49-1.78ppm(m,3H).

[0380] Example 22

[0381]

[0382] 2-((3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenyl)amino 2-methylpropionic acid

[0383] CuI (1.00 mg, 0.021 mmol), pyrrolidine-2-carboxylic acid (1.2 mg, 0.042 mmol), and Cs₂CO₃ (137.11 mg, 0.421 mmol) were added to a suspension of 4-bromo-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (80 mg, 0.210 mmol) and 2-amino-2-methylpropionic acid (0.04 mL, 0.421 mmol) in DMSO (2 mL) under N₂ conditions, and the mixture was stirred at 110 °C for 16 hours. The mixture was a yellow solution. The mixture was filtered and washed with DMSO (1 mL). The filtrate was purified by preparative HPLC (column: YMC-Actus Triart C18 150*30mm*7um; mobile phase: [water (0.225% FA)-ACN]; B%: 23%-63%, 10 min) to give the title compound (19 mg, yield: 21%) as a white solid. LC-MS (ESI): m / z 403.1 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ10.60(br s,1H),7.93(d,J=5.1Hz,1H),7.77(d,J=3.8Hz,1H),7.64(d,J=8.9Hz,1H),7.29(t,J=4.5Hz,1H),6.28-6.14(m,2H),3.86(s,3H),1.48(s,6H).

[0384] Example 23

[0385]

[0386] 4-(6-oxa-3-azabicyclo[3.1.1]hept-3-yl)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4- Oxadiazol-2-yl)benzamide

[0387] RuPhos-Pd-G3 (144 mg, 0.17 mmol) was added to a solution of 4-bromo-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (65 mg, 0.17 mmol), 6-oxa-3-azabicyclo[3.1.1]heptane 4-methylbenzenesulfonate TSOH salt (70 mg, 0.26 mmol) and Cs₂CO₃ (392 mg, 1.2 mmol) in tert-amyl alcohol (2 mL) at 25 °C under N₂. The mixture was stirred at 120 °C for 16 hours. The solvent was removed under vacuum, and the residue was purified by preparative HPLC (column: Ultimate C18 100*30mm*3um; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN; B%: 41%-81%, 10 min)) to give the title compound (17 mg, yield: 24%) as a yellow solid. LC-MS (ESI): m / z 399.0 [M+H) + . 1 H NMR (DMSO-d6, 400MHz): δ7.65(d,J=5.0Hz,1H),7.52(d,J=8.3Hz,1H),7.45(d,J=3.6Hz,1H),7.13-7.20(m,1H),6.12-6.36(m,2H),4.71(br d,J=6.3Hz,2H),3.73(s,3H),3.57(br d,J=11.4Hz,2H),3.44(br,s,2H),3.07-3.17(m,1H),2.08(s,1H),1.93(d,J=8.8Hz,1H).

[0388] Example 24

[0389]

[0390] 2-Methoxy-4-morpholino-N-(5-(thiazolyl-5-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0391] a) Methyl 2-methoxy-4-morpholinobenzoate

[0392] A solution of methyl 4-fluoro-2-methoxybenzoate (10 g, 54 mmol) in morpholine (38 mL, 434 mmol) and NMP (30 mL) was prepared at 25 °C under N2. The mixture was stirred at 145 °C for 16 hours. The mixture was freeze-dried to remove the solvent, giving the residue. The residue was then analyzed by rapid silica gel chromatography (…). 120g The residue was purified by silica gel rapid column chromatography (eluting with a gradient of 0–30% ethyl acetate / petroleum ether = 1:2) to give the title compound (3.5 g, yield: 18%) as a white solid. LC-MS (ESI): m / z 252.0 [M+H] + .

[0393] b) 2-Methoxy-4-morpholinobenzoic acid

[0394] LiOH·H₂O (12.59 mg, 0.3 mmol) was added in a fraction of a solution of methyl 2-methoxy-4-morpholinobenzoate (3.0 g, 8.5 mmol) in a mixed solvent of THF (40 mL) and H₂O (40 mL) at 25 °C. The reaction mixture was heated to 40 °C for 16 hours. The solvent was removed, and the residue was acidified to pH 2 with 1 N HCl. A solid was formed and collected by filtration. The solid was washed with H₂O (10 mL × 3) and dried under vacuum to give the title compound (1.2 g, yield: 52%) as a white solid. LC-MS (ESI): m / z 238.1 [M+H] + .

[0395] c) 2-Methoxy-4-morpholino-N-(5-(thiazo-5-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0396] T3P (1.50 mL, 5.1 mmol) and TEA (0.70 mL, 5.1 mmol) were added to a suspension of 2-methoxy-4-morpholinobenzoic acid (300 mg, 1.3 mmol) and 5-(thiazol-5-yl)-1,3,4-oxadiazol-2-amine (319 mg, 1.9 mmol) in EtOAc (2 mL) at 25 °C. The reaction mixture was heated at 80 °C for 16 hours. The mixture was diluted with H2O (10 mL) and extracted with EtOAc (10 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was wet-milled with DMSO (4 mL), filtered, and washed with H2O (5 mL × 3) to give a crude product. The crude product was washed with MTBE (5 mL) to give the title compound (110 mg, yield: 20%) as a yellow solid. LC-MS(ESI): m / z 388.1 [M+H] + . 1 HNMR(DMSO-d6,400MHz): δ10.87(s,1H),9.40(s,1H),8.56(s,1H),7.73(d,J=8.9Hz,1H),6 .67(dd,J=8.8,1.8Hz,1H),6.59(s,1H),3.96(s,3H),3.69-3.80(m,4H),3.35-3.38(m,4H).

[0397] Example 25

[0398]

[0399] 2-Methoxy-4-morpholino-N-(5-(thiazolyl-5-yl)-1,3,4-oxadiazol-2-yl)benzenesulfonamide

[0400] NaH (36 mg, 0.9 mmol, 60% purity, in mineral oil) was added in a one-part ratio to a suspension of 5-(thiazolyl-5-yl)-1,3,4-oxadiazol-2-amine (50 mg, 0.3 mmol) in THF (3 mL), and the reaction mixture was stirred at 0 °C for 15 min. 2,4-Dimethoxybenzenesulfonyl chloride (120 mg, 0.5 mmol) was added in a one-part ratio, and the reaction mixture was stirred at 25 °C for 16 h. H₂O (1 mL) was added to the reaction mixture at 0 °C to quench the reactants, and the reaction mixture was then concentrated under reduced pressure to give the residue. The residue was purified by preparative HPLC (column: Daisogel SP ODS RPS 150*25 mm*5 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 18%-58%, 11 min) to give the title compound (56 mg, yield: 51%) as a white solid. LC-MS(ESI): m / z 369.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ9.38 (s, 1H), 8.48 (s, 1H), 7.78 (d, J = 8.7Hz, 1H), 6.60-6.70 (m, 2H), 3.83 (s, 3H), 3.75 (s, 3H).

[0401] Example 26

[0402]

[0403] 2-Methoxy-N-methyl-4-morpholino-N-(5-(thiazolyl-5-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0404] a) 2,4-Dimethoxy-N-(5-(thiazo-5-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0405] 2,4-Dimethoxybenzoic acid (162.48 mg, 0.90 mmol) and TEA (0.33 mL, 2.38 mmol) were added in a one-part ratio to a suspension of 5-(thiazol-5-yl)-1,3,4-oxadiazol-2-amine (100 mg, 0.60 mmol) in EtOAc (0.6 mL) at 25 °C, and stirred for 10 min. Then, T3P (1.51 g, 2.38 mmol, 1.41 mL, 50% purity) was added, and the reaction mixture was heated to 80 °C and stirred at 80 °C for 16 h, resulting in a brown solution. The reaction mixture was cooled to 25 °C, water (2 mL) was added to the reaction mixture, and the mixture was extracted with EtOAc (1 mL × 2). The combined organic layers were separated, dried over Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Daisogel SP ODS RPS 150*25mm*5um; mobile phase: [water (0.225% FA)-ACN]; B%: 28%-68%, 30 min) to give the title compound (0.1 g, yield: 51%) as a white solid. LC-MS (ESI): m / z 333.1 [M+H] + .

[0406] b) 2,4-Dimethoxy-N-methyl-N-(5-(thiazo-5-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0407] K₂CO₃ (16.59 mg, 0.120 mmol) was added in a single-part batch to a solution of 2,4-dimethoxy-N-(5-(thiazolyl-5-yl)-1,3,4-oxadiazol-2-yl)benzamide (10 mg, 0.030 mmol) in DMF (0.3 mL) at 25 °C, and the mixture was stirred for 10 min. Then CH₃I (3.75 μL, 0.060 mmol) was added, and the reaction mixture was heated to 55 °C and stirred at 55 °C for 30 min. The reaction mixture was filtered, and the filtrate was purified by preparative HPLC (column: YMC-Triart Prep C18 150*40 mm*7 μm; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN; B%: 25%-65%, 11 min) to give the title compound (1.5 mg, yield: 14%) as a white solid. LC-MS(ESI): m / z 347.1 [M+H] + . 1 H NMR (CDCl3, 400MHz) δ8.92 (s, 1H), 8.03 (s, 1H), 7.56 (d, J = 8.4Hz, 1H), 6.62 (d d,J=8.4,1.8Hz,1H),6.35(s,1H),3.87(s,3H),3.66(s,3H),3.59ppm(s,3H).

[0408] Example 27

[0409]

[0410] 2-Methoxy-4-morpholino-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide

[0411] T3P (75 mL, 0.25 mmol) and TEA (35 mL, 0.25 mmol) were added to a suspension of 2-methoxy-4-morpholinobenzoic acid (40 mg, 0.17 mmol) and 5-(thiophen-2-yl)-1,3,4-oxadiazol-2-amine (42 mg, 0.25 mmol) in ethyl acetate (0.4 mL) at 25 °C, and the reaction mixture was stirred at 80 °C for 16 h. The mixture was diluted with H2O (3 mL) and extracted with ethyl acetate (3 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under reduced pressure. The residue was purified by preparative HPLC (column: Phenomenex Gemini-NX 150*30 mm*5 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 26%-66%, 10 min) to give the title compound (17 mg, yield: 25%) as a yellow solid. LC-MS(ESI): m / z 387.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ10.79 (br s, 1H), 7.92 (br d, J = 4.8Hz, 1H), 7.71-7.82 (m, 2H), 7.29 (t, J = 4.3Hz, 1H), 6.66 (br d,J=8.5Hz,1H),6.57(s,1H),3.95(s,3H),3.73(br d,J=4.5Hz,4H),3.55-3.57(m,4H).

[0412] Example 28

[0413]

[0414] 2,4-Dimethoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide

[0415] a)(2-oxo-2-(2-(thiophene-2-carbonyl)hydrazino)ethyl)tert-butyl carbamate

[0416] HATU (1.93 g, 5.07 mmol) and DIEA (1.94 mL, 11.7 mmol) were added to a suspension of thiophene-2-carboxylic acid (0.50 g, 3.90 mmol) and (2-oxo-2-(2-(thiophene-2-carbonyl)hydrazino)ethyl)carbamate (0.96 g, 5.07 mmol) in DMF (40 mL) at 25 °C, and the reaction mixture was stirred at 25 °C for 16 hours. The solvent was removed from the mixture under vacuum to give the title compound (2.7 g, 81%) as a yellow oil, which was used directly in the next step without purification. LC-MS (ESI): m / z 244.1 [M+H] + .

[0417] b)((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl)tert-butyl carbamate

[0418] Triethylamine (1.31 mL, 9.47 mmol) was added to a solution of tert-butyl (2.7 g, 3.16 mmol) and PPh3 (2.48 g, 9.47 mmol) in acetonitrile (50 mL), and the mixture was stirred at 25 °C for 20 min. Carbon tetrachloride (0.92 mL, 9.47 mmol) was added to the mixture, and the mixture was stirred at 25 °C for 16 h. The solvent was removed under vacuum. The final product was analyzed by rapid silica gel chromatography (…). 40g The residue was purified by a silica gel fast column chromatography (eluting with a gradient of 0–50% ethyl acetate / petroleum ether) to give the title compound (800 mg, 81%) as a pale yellow solid. LC-MS (ESI): m / z 282.1 [M+H] + .

[0419] c)(5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methylamine TFA salt

[0420] TFA (1.5 mL) was added dropwise to a solution of ((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)carbamate (150 mg, 0.48 mmol) in DCM (1.5 mL) at 25 °C. The mixture was stirred at 25 °C for 2 hours. The solvent was removed under vacuum and N2 to give the title compound (130 mg, 0.47 mmol, 94%) as a pale yellow solid. LC-MS (ESI): m / z 182.2 [M+H] + .

[0421] d) 2,4-Dimethoxy-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide

[0422] T3P (0.6 mL, 1.0 mmol) and TEA (0.3 mL, 2.0 mmol) were added to a suspension of (5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methylamine TFA salt (65 mg, 0.25 mmol) and 2,4-dimethoxybenzoic acid (68.61 mg, 0.38 mmol) in EtOAc (2 mL) at 25 °C. The mixture was stirred at 80 °C for 16 h. The mixture was diluted with H2O (3 mL) and extracted with ethyl acetate (5 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under vacuum. The residue was purified by preparative HPLC (column: Boston Green ODS 150*30 mm*5 μm; mobile phase: [water (0.225% FA)-ACN]; B%: 26%-66%, 10 min) to give the title compound (17.8 mg, 21%) as a white solid. LC-MS(ESI): m / z 346.1 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ8.77(t,J=5.7Hz,1H),7.93(dd,J=5.0,1.1Hz,1H),7.85(d,J=8.6Hz,1H),7.78(dd,J=3 .7,1.1Hz,1H),7.28(dd,J=4.9,3.8Hz,1H),6.58-6.73(m,2H),4.76(d,J=5.6Hz,2H),3.94(s,3H),3.83(s,3H).

[0423] The following compounds were synthesized according to the procedure described in Example 28.

[0424]

[0425]

[0426]

[0427] Example 29

[0428]

[0429] 2-Methoxy-4-morpholino-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide

[0430] a)(5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methylamine HBF4 salt

[0431] HBF4 (0.02 mL, 0.16 mmol) was added dropwise to a solution of ((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)carbamate tert-butyl ester (50 mg, 0.16 mmol) in TFE (0.5 mL) at 0 °C. The mixture was stirred at 0 °C for 15 minutes. The mixture was dissolved in H2O (2 mL) at 0 °C and freeze-dried to give the title compound (43 mg, yield: 73%) as a pale yellow solid. LC-MS (ESI): m / z 182.2 [M+H] + .

[0432] b) 2-Methoxy-4-morpholino-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide

[0433] T3P (0.2 mL, 0.33 mmol) and TEA (0.1 mL, 0.66 mmol) were added to a suspension of 2-methoxy-4-morpholinobenzoic acid (19 mg, 0.083 mmol) and (5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methylamine (45 mg, 0.12 mmol) in ethyl acetate (0.5 mL) at 25 °C, and the reaction mixture was stirred at 80 °C for 16 hours. The mixture was diluted with H2O (2 mL) and extracted with ethyl acetate (3 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under vacuum to give the residue. The residue was purified by preparative HPLC (column: YMC-Actus Triart C18 150*30mm*7um; mobile phase: [water (0.225% FA)-ACN]; B%: 23%-63%, 10 min) to give the title compound (4.7 mg, yield: 14%) as a white solid. LC-MS (ESI): m / z 401.0 [M+H] + . 1 H NMR (DMSO-d6, 400MHz): δ8.69 (t, J=5.4Hz, 1H), 7.94 (dd, J=5.0, 1.0Hz, 1H), 7.71-7.85 (m, 2H), 7.28 (dd, J=4. 9,3.8Hz,1H),6.47-6.73(m,2H),4.75(d,J=5.6Hz,2H),3.95(s,3H),3.70-3.81(m,4H),3.22-3.30ppm(m,4H).

[0434] Example 30

[0435]

[0436] 2-Methoxy-N-methyl-4-morpholino-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzene formamide

[0437] NaH (60%, 15.0 mg, 0.38 mmol) was added to a solution of 2-methoxy-4-morpholino-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (100 mg, 0.25 mmol) in THF (1.5 mL) at 0 °C under N2, and the mixture was stirred for 0.5 h. Iodomethane (78 mL, 1.25 mmol) was added to the mixture at 0 °C under N2. The mixture was stirred at 25 °C for 1 h. The mixture was diluted with H2O (5 mL) and extracted with EtOAc (5 mL × 3). The organic layer was separated, dried over MgSO4, filtered, and the filtrate was concentrated under vacuum. The residue was purified by preparative HPLC (column: YMC Triart C18 150×25mm×5um; mobile phase: [water (0.225% FA)-ACN]; B%: 35%-55%, 10 min) to give the title compound (14.1 mg, 13.6%) as a white solid. LC-MS (ESI): m / z 415.2 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ7.97(d,J=4.8Hz,1H),7.86-7.73(m,1H),7.35-7.24(m,1H),7.06(d,J=8.3Hz,1H),6 .56(d,J=11.5Hz,2H),4.94(s,1H),4.64(s,1H),3.81-3.64(m,7H),3.19(d,J=3.5Hz,4H),3.09-2.86(m,3H).

[0438] The following compounds were synthesized according to the procedure described in Example 30.

[0439]

[0440] Example 31

[0441]

[0442] 2-Methoxy-3-morpholino-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide

[0443] a) Methyl 2-methoxy-3-morpholinobenzoate

[0444] A mixture of methyl 3-bromo-2-methoxybenzoate (1.10 g, 4.49 mmol), morpholine (0.79 mL, 8.98 mmol), Pd2(dba)3 (411 mg, 0.45 mmol), Cs2CO3 (4.39 g, 13.5 mmol), BINAP (559 mg, 0.898 mmol), and toluene (30 mL) was degassed three times with argon and heated to 100 °C for 6 hours. The mixture was then mixed with water (100 mL) and extracted with EA (50 mL × 2). The organic layers were combined, washed with brine (50 mL × 3), dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (PE / EA = 100 / 1-4 / 1) to give the title compound (303 mg, yield: 26.9%) as a yellow oil. LC-MS(ESI): m / z 252.1 [M+H] + .

[0445] b) 2-Methoxy-3-morpholinobenzoic acid

[0446] LiOH·H₂O (442 mg, 10.6 mmol) was added to a solution of methyl 2-methoxy-3-morpholinobenzoate (331 mg, 1.32 mmol) in MeOH (15 mL) and H₂O (15 mL), and the resulting mixture was heated to 80 °C for 2 hours. The mixture was concentrated under reduced pressure to remove most of the MeOH, and the pH of the resulting mixture was adjusted to 1–2 with an aqueous HCl solution (1.0 M). The resulting suspension was filtered under vacuum. The residue was slurried with (EA / PE = 1 / 10, 44 mL) to give the title compound (89 mg, yield: 29%) as a white solid. LC-MS (ESI): m / z 238.2 [M+H] + .

[0447] c) 2-Methoxy-3-morpholino-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide

[0448] Add (5-(thiophene-2-yl)-1,3,4-oxadiazol-2-yl)methylamine (20 mg, 0.11 mmol), DIEA (0.06 mL, 0.39 mmol), and HATU (44 mg, 0.12 mmol) to a solution of 2-methoxy-3-morpholinobenzoic acid (52 mg, 0.22 mmol) in DMF (20 mL). Stir the reaction mixture at room temperature for 30 minutes. Dilute the resulting mixture with water (50 mL) and extract with ethyl acetate (50 mL × 2). Combine the organic layers, wash with brine (50 mL × 3), dry to anhydrous Na₂SO₄, and filter. Concentrate the filtrate under reduced pressure. The residue was purified by preparative HPLC (method: Waters 2767 / 2545 / 2489 / Qda, Waters Xbridge C18 10µm OBD 19*150mm, mobile phase A: water containing 0.1% NH4HCO3, mobile phase B: CH3CN, flow rate: 20mL / min, column temperature: room temperature) to give the title compound (15mg, yield: 34%) as a white solid. LC-MS (ESI): m / z 401.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.98(t,J=5.4Hz,1H),7.94(d,J=4.8Hz,1H),7.79(d,J=3.0Hz,1H),7.27–7.32(m,1H) ,7.25(d,J=5.8Hz,1H),7.08–7.17(m,2H),4.76(d,J=5.6Hz,2H),3.86(s,3H),3.77(s,4H),3.00–3.07(m,4H).

[0449] Example 32

[0450]

[0451] 4-(2,6-Dimethylmorpholinyl)-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl (Bento) benzamide

[0452] a) Methyl 4-(2,6-dimethylmorpholino)-2-methoxybenzoate

[0453] A suspension of methyl 4-fluoro-2-methoxybenzoate (500 mg, 2.715 mmol), 2,6-dimethylmorpholine (0.50 mL, 4.07 mmol), and K₂CO₃ (563 mg, 4.07 mmol) in DMSO (10 mL) was heated at 120 °C for 16 hours. The resulting mixture was diluted with water (40 mL) and extracted with EtOAc (20 mL × 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under reduced pressure. The residue was purified by silica gel chromatography (PE / EA = 20 / 1–5 / 1) to give the title compound (115 mg, 15% yield) as a white solid. LC-MS (ESI): m / z 280.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.62 (d, J = 8.8 Hz, 1H), 6.55 (dd, J = 8.9, 2.1 Hz, 1H), 6.51 (s, 1H), 2.42–2.31 (m, 2H), 1.17 (d, J = 6.2 Hz, 6H).

[0454] b) 4-(2,6-dimethylmorpholino)-2-methoxybenzoic acid

[0455] A mixture of methyl 4-(2,6-dimethylmorpholino)-2-methoxybenzoate (115 mg, 0.412 mmol), LiOH·H₂O (34.6 mg, 0.823 mmol), THF (5.0 mL), and water (2.5 mL) was heated at 70 °C for 8 hours. The resulting mixture was adjusted to pH 5–6 with dilute HCl (1.0 M) and extracted with EtOAc (10 mL × 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous Na₂SO₄, and filtered under vacuum. The filtrate was concentrated under reduced pressure to give the title compound (100 mg, yield: 91%) as a pale yellow solid. LC-MS (ESI): m / z 266.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.69(d,J=8.7Hz,1H),6.60(d,J=9.0Hz,1H),6.57(s,1H),3.88(s,3H) ,3.83(d,J=12.1Hz,3H), 3.71(d,J=6.7Hz,3H), 2.42(t,J=11.3Hz,2H), 1.23(d,J=6.1Hz,5H).

[0456] c) 4-(2,6-Dimethylmorpholino)-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl) Methylbenzamide

[0457] A mixture of 4-(2,6-dimethylmorpholino)-2-methoxybenzoic acid (100 mg, 0.377 mmol), DIEA (72.9 mg, 0.565 mmol), and HATU (172 mg, 0.452 mmol) in DMF (3.0 mL) was stirred at room temperature for 30 min, followed by the addition of (5-(thiophene-2-yl)-1,3,4-oxadiazol-2-yl)methylamine TFA salt (68.3 mg, 0.377 mmol). The resulting reaction mixture was stirred at room temperature for 1 h. The resulting mixture was diluted with water (15 mL) and extracted with EA (30 mL × 3). The organic layers were combined, washed with brine (30 mL), dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated under reduced pressure. The residue was purified by silica gel chromatography (PE / EA = 10 / 1 to 1 / 1) to give the title compound (138 mg, yield: 85%) as a white solid. LC-MS(ESI): m / z 429.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.66(d,J=5.5Hz,1H),7.93(d,J=4.9Hz,1H),7.76(d,J=8.7Hz,2H),7.31–7.25(m,1H),6.61(d,J=8.9Hz,1H) ,6.56(s,1H),4.75(d,J=5.5Hz,2H),3.95(s,3H),3.78(d,J=11.9Hz,2H),3.66(s,2H),2.36(t,J=11.3Hz,2H),1.17(d,J=6.1Hz,6H).

[0458] Example 33

[0459]

[0460] 2-Methoxy-4-(piperidin-1-yl)-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzyl amide

[0461] a) 4-Bromo-2-methoxy-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide

[0462] EDC·HCl (95.2 mg, 0.40 mmol) and DMAP (4.0 mg, 0.033 mmol) were added to a suspension of 4-bromo-2-methoxybenzoic acid (92.0 mg, 0.40 mmol) and (5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methylamine (100 mg, 0.331 mmol) in pyridine (2 mL) at 25 °C, and the reaction mixture was stirred at 50 °C for 2 hours. The mixture was concentrated under vacuum. The residue was slurried with acetonitrile (3 mL) and H₂O (0.5 mL) to give the title compound (80 mg, 0.18 mmol, 55.4%) as a white solid. LC-MS (ESI): m / z 394.0 [M+H] + .

[0463] b) 2-Methoxy-4-(piperidin-1-yl)-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzene formamide

[0464] RuPhos-Pd-G3 (42.0 mg, 0.05 mmol) was added to a solution of 4-bromo-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (50 mg, 0.12 mmol), piperidine (10 mL, 0.1 mmol), and Cs₂CO₃ (65.4 mg, 0.20 mmol) in tert-amyl alcohol (5 mL). The mixture was stirred at 120 °C for 16 hours. The mixture was diluted with H₂O (5 mL) and extracted with EtOAc (5 mL × 3). The organic layer was separated, dried over MgSO₄, filtered, and the filtrate was concentrated under vacuum. The residue was purified by preparative HPLC (column: YMC Triart C18 150×25mm×5um; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 43%-63%, 10 min) to give the title compound (9 mg, 21%) as a white solid. LC-MS (ESI): m / z 399.2 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ7.93(d,J=5.0Hz,1H),7.81-7.70(m,2H),7.32-7.22(m,1H), 6.93(s,1H),6.61-6.46(m,2H),4.75(s,2H),3.94(s,3H),3.30(s,4H),1.59(s,6H).

[0465] The following compounds were synthesized according to the procedure described in Example 33.

[0466]

[0467] Example 34

[0468]

[0469] 2-Methoxy-4-morpholino-N-((5-(thiazolyl-5-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide

[0470] a)(2-oxo-2-(2-(thiazo-5-carbonyl)hydrazino)ethyl)tert-butyl carbamate

[0471] HOBt (0.09 g, 0.698 mmol), EDC (0.35 g, 1.816 mmol), and (tert-butyloxycarbonyl)glycine (0.88 g, 4.396 mmol) were added to a suspension of thiazolium-5-carbonylhydrazine (0.2 g, 1.4 mmol) in THF (5.6 mL) and DMF (1.4 mL) at 25 °C, and the mixture was stirred for 16 hours. The mixture was diluted with 1N NaOH (18 mL) and extracted with DCM (15 mL × 3). The aqueous layer was dehydrated under vacuum to give the title compound (4 g, 95%) as a yellow solid. LC-MS (ESI): m / z 301.1 [M+H] + .

[0472] b)((5-(thiazolyl-5-yl)-1,3,4-oxadiazol-2-yl)methyl)tert-butyl carbamate

[0473] TEA (0.5 mL, 3.3 mmol) was added to a solution of (2-oxo-2-(2-(thiazolyl-5-carbonyl)hydrazino)ethyl)carbamate (3.3 g, 1.1 mmol) and PPh3 (0.86 g, 3.3 mmol) in acetonitrile (6 mL). The reaction mixture was stirred at 25 °C for 20 min. CCl4 (0.3 mL, 3.3 mmol) was added to the mixture, and the mixture was stirred at 25 °C for 16 h. The solvent was removed under vacuum. The residue was diluted with H2O (5 mL) and extracted with ethyl acetate (10 mL × 3). The combined layers were dried over MgSO4, filtered, and the filtrate was concentrated under vacuum. The filtrate was analyzed by rapid silica gel chromatography (…). 40g The residue was purified by a silica gel fast column chromatography (eluting with a gradient of 0–30% ethyl acetate / petroleum ether) to give the title compound (0.6 g, 97%) as a white solid. LC-MS (ESI): m / z 283.1 [M+H] + .

[0474] c)((5-(thiazolyl-5-yl)-1,3,4-oxadiazol-2-yl)methyl)tert-butyl carbamate HBF4 salt

[0475] HBF4 (0.14 mL, 1.06 mmol) was added dropwise to a solution of ((5-(thiazol-5-yl)-1,3,4-oxadiazol-2-yl)methyl)carbamate tert-butyl ester (0.6 g, 1.1 mmol) in TFE (6 mL) at 0 °C. The mixture was stirred at 25 °C for 2 hours. The white suspension was dissolved in H2O (5 mL) at 0 °C and freeze-dried. The solid was slurried with DCM (1 mL) to give the title compound (47 mg, 17%) as a yellow solid. LC-MS (ESI): m / z 183.1 [M+H] + .

[0476] d) 2-Methoxy-4-morpholino-N-((5-(thiazolyl-5-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide

[0477] EDC·HCl (52 mg, 0.27 mmol) and DMAP (2.21 mg, 0.018 mmol) were added to a suspension of ((5-(thiazolyl-5-yl)-1,3,4-oxadiazol-2-yl)methyl)carbamate tert-butyl ester HBF4 salt (57.12 mg, 0.217 mmol) and 2-methoxy-4-morpholinobenzoic acid (47 mg, 0.181 mmol) in pyridine (1 mL) at 25 °C. The reaction mixture was stirred at 50 °C for 2 h. The mixture was a yellow solution. The solvent was removed from the mixture under vacuum. The residue was purified by preparative HPLC (column: YMCTriart C18 150*25 mm*5 mm; mobile phase: [water (0.05% ammonium hydroxide v / v)-ACN]; B%: 25%-45%, 10 min) to give the title compound (36 mg, 49%) as a white solid. LC-MS(ESI): m / z 402.1 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ9.40(s,1H),8.69(t,J=5.5Hz,1H),8.56(s,1H),7.78(d,J=8.8Hz,1H) ,6.69-6.51(m,2H),4.77(d,J=5.5Hz,2H),3.95(s,3H),3.78-3.70(m,4H),3.29-3.25(m,4H).

[0478] The following compounds were synthesized according to the procedure described in Example 34.

[0479]

[0480]

[0481] Example 35

[0482]

[0483] N-((5-(5-chlorothiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)-2-methoxy-4-morpholinylbenzoyl amine

[0484] a)((5-(5-chlorothiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)tert-butyl carbamate

[0485] NCS (36 mg, 0.3 mmol) was added to a mixture of ((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)carbamate tert-butyl ester (50 mg, 0.18 mmol) in DMF (0.8 mL). The mixture was stirred at 20 °C under N2 for 16 h. The mixture was diluted with 1 M NaOH (3 mL) and extracted with EtOAc (3 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated to give the title compound (56 mg, yield: 99%) as a yellow oil. LC-MS (ESI): m / z 316.1 [M+H] + .

[0486] b)(5-(5-chlorothiophen-2-yl)-1,3,4-oxadiazol-2-yl)methylamine HBF 4 Salt

[0487] HBF4 (37 μL, 0.28 mmol) was added dropwise to a solution of ((5-(5-chlorothiophene-2-yl)-1,3,4-oxadiazol-2-yl)methyl)carbamate tert-butyl ester (87 mg, 0.28 mmol) in TFE (1 mL) at 0 °C. The mixture was stirred at 25 °C for 2 hours. The mixture was a white suspension. The mixture was dissolved in H2O (2 mL) at 0 °C and freeze-dried to give the title compound (60 mg, yield: 91%) as a pale yellow solid. LC-MS (ESI): m / z 216.0 [M+H] + .

[0488] c)N-((5-(5-chlorothiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)-2-methoxy-4-morpholinylbenzyl amide

[0489] EDC·HCl (72 mg, 0.3 mmol) and DMAP (3 mg, 0.03 mmol) were added to a suspension of 2-methoxy-4-morpholinobenzoic acid (79 mg, 0.3 mmol) and (5-(5-chlorothiophene-2-yl)-1,3,4-oxadiazol-2-yl)methylamine (60 mg, 0.3 mmol) in pyridine (1.2 mL) at 25 °C, and the reaction mixture was stirred at 50 °C for 2 hours. The solvent was removed under vacuum. The residue was slurried in MeCN (3 mL) and DMSO (1 mL). The solid was collected by filtration to give the title compound (47 mg, yield: 41%) as a white solid. LC-MS (ESI): m / z 435.1 [M+H]+ . 1 H NMR (400MHz, DMSO-d6) δ8.68(t,J=5.6Hz,1H),7.77(d,J=8.8Hz,1H),7.66(d,J=4.0Hz,1H),7.34(d,J=4.0Hz ,1H),6.65-6.50(m,2H),4.80-4.65(m,2H),3.94(s,3H),3.77-3.70(m,4H),3.30-3.25(m,4H),1.99(s,1H).

[0490] Example 36

[0491]

[0492] 2-Methoxy-4-morpholino-N-(2-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)propyl-2-yl)benzyl amide

[0493] a)(2-methyl-1-oxo-1-(2-(thiophene-2-carbonyl)hydrazino)prop-2-yl)tert-butyl carbamate

[0494] HOBt (0.24 g, 1.8 mmol), EDC (0.84 g, 4.4 mmol), and 2-((tert-butoxycarbonyl)amino)-2-methylpropionic acid (0.88 g, 4.4 mmol) were added to a suspension of thiophene-2-carbonylhydrazine (0.5 g, 3.5 mmol) in THF (14 mL) and DMF (4 mL) at room temperature. The reaction mixture was stirred for 16 hours. The mixture was diluted with 1 M NaOH (18 mL) and extracted with DCM (15 mL × 3). The combined organic layers were dried over MgSO4, filtered, and the solvent was removed under vacuum to give the title compound (1.1 g, 48%) as a yellow oil. LC-MS (ESI): m / z 328.1 [M+H] + .

[0495] b)(2-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)propyl-2-yl)tert-butyl carbamate

[0496] Triethylamine (318 μL, 2.3 mmol) was added to a solution of tert-butyl (2-methyl-1-oxo-1-(2-(thiophene-2-carbonyl)hydrazino)prop-2-yl)carbamate (500 mg, 0.76 mmol) and PPh3 (600 mg, 2.3 mmol) in acetonitrile (5 mL). The reaction mixture was stirred at 25 °C for 20 min. CCl4 (221 μL, 2.3 mmol) was added to the mixture, and the mixture was stirred at 25 °C for 16 h. The solvent was removed under vacuum, and the mixture was analyzed by rapid silica gel chromatography (…). 40g The residue was purified by silica gel fast column chromatography (eluting with a gradient of 0–50% ethyl acetate / petroleum ether) to give the title compound (100 mg, 35%) as a pale yellow solid. LC-MS (ESI): m / z 310.0 [M+H] + .

[0497] c) 2-(5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)propyl-2-amine

[0498] HBF4 (44 μL, 0.33 mmol) was added dropwise to a solution of tert-butyl (2-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)propyl-2-yl)carbamate (100 mg, 0.32 mmol) in TFE (1 mL) at 0 °C. The mixture was stirred at 25 °C for 2 hours. The mixture was dissolved in H2O (2 mL) and freeze-dried to give the title compound (90 mg, 93%) as a yellow solid. LC-MS (ESI): m / z 210.0 [M+H] + .

[0499] d) 2-Methoxy-4-morpholino-N-(2-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)propyl-2-yl)benzene formamide

[0500] At room temperature, EDC·HCl (58 mg, 0.30 mmol) and DMAP (2.5 mg, 0.02 mmol) were added to a suspension of 2-methoxy-4-morpholinobenzoic acid (63 mg, 0.241 mmol) and 2-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)propyl-2-amine (60 mg, 0.20 mmol) in pyridine (1.2 mL). The reaction mixture was stirred at 50 °C for 2 hours. The solvent was removed under vacuum. The residue was slurried in acetonitrile (3 mL) and DMSO (1 mL), and the solid was collected by filtration to give the title compound (18 mg, yield: 20%) as a white solid. LC-MS (ESI): m / z 429.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.39(s,1H),7.91(dd,J=1.1,5.0Hz,1H),7.72(dd,J=1.1,3.6Hz,1H),7.63(d,J=9.4Hz,1H ),7.25(dd,J=3.8,4.9Hz,1H),6.62-6.55(m,2H),3.99(s,3H),3.77-3.71(m,4H),3.28-3.24(m,4H),1.76(s,6H).

[0501] The following compounds were synthesized according to the procedure described in Example 36.

[0502]

[0503] Example 37

[0504]

[0505] 2-Methoxy-N-methyl-4-morpholinyl-N-(2-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)prop-2- (Bento) benzamide

[0506] NaH (60% in mineral oil, 8.40 mg, 0.21 mmol) was added to a solution of 2-methoxy-4-morpholino-N-(2-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)propyl-2-yl)benzamide (400 mg, 0.14 mmol) in THF (1 mL) at 0 °C under N2, and the mixture was stirred for 0.5 h. Iodimethane (44 μL, 0.7 mmol) was added to the mixture at 0 °C under N2. The mixture was stirred at 25 °C for 1 h. The mixture was diluted with H2O (1 mL) and extracted with ethyl acetate (1 mL × 3). The combined organic layers were dried over MgSO4, filtered, and concentrated under vacuum. The residue was purified by preparative HPLC (column: YMCTriart C18 150*25mm*5mm; mobile phase: [water (0.225% FA)-ACN]; B%: 35%-55%, 10 min) to give the title compound (4.74 mg, 6.6%) as a white solid. LC-MS (ESI): m / z 443.2 [M+H) + . 1 HNMR(400MHz,DMSO-d6)δ8.43(br s,1H),7.92(dd,J=1.2,5.1Hz,1H),7.73(dd,J=1.2,3.7Hz,1H),7.29(dd,J=3.7,5.0Hz,1H),6.97(d,J=8. 4Hz,1H),6.64-6.44(m,2H),3.83(s,3H),3.77-3.68(m,4H),3.26-3.11(m,4H),2.92(s,3H),1.72(s,6H).

[0507] Example 38

[0508]

[0509] 2-Methoxy-4-(2-morpholinylethoxy)-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl) benzamide

[0510] a) 4-Hydroxy-2-methoxybenzoic acid

[0511] NaH₂PO₄ (394 mg, 161.025 mmol) and NaClO₂ (11.7 g, 129 mmol) were added to a solution of 4-hydroxy-2-methoxybenzaldehyde (4.9 g, 32.2 mmol) in DMSO (100 mL) and water (75 mL) at 0 °C. The reaction mixture was then stirred at room temperature for 3 hours. The mixture was mixed with water (400 mL) and the pH was adjusted to 8–9 with NaHCO₃ powder. The entire mixture was washed with EA (300 mL). The aqueous phase was adjusted to 3–4 with an aqueous HCl solution (1.0 M) and the resulting mixture was extracted with ethyl acetate (100 mL × 3). The combined organic layers were washed with brine (200 mL), dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated under reduced pressure to give the title compound (5.39 g, yield: 97.1%) as a yellow oil. LC-MS (ESI): m / z 168.8 [M+H] + .

[0512] b) 4-Hydroxy-2-methoxy-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide

[0513] BOP (3.95 g, 8.92 mmol), DIEA (3.94 mL, 23.8 mmol), and (5-(thiophene-2-yl)-1,3,4-oxadiazol-2-yl)methylamine (1.82 g, 6.54 mmol) were added to a solution of 4-hydroxy-2-methoxybenzoic acid (1.00 g, 5.95 mmol) in DMF (20 mL), and the reaction mixture was stirred at room temperature for 1.5 h. The reaction mixture was diluted with water (20 mL). The mixture was extracted with ethyl acetate (20 mL × 2). The organic phases were combined, washed with brine (30 mL × 2), dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated under reduced pressure to give the title compound (1.8 g, yield: 63.9%) as a yellow solid. LC-MS (ESI): m / z 332.0 [M+H] + .

[0514] c) 2-Methoxy-4-(2-morpholinylethoxy)-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl (Bento) benzamide

[0515] To a solution of 4-hydroxy-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (200 mg, 0.42 mmol) in acetonitrile (3.0 mL), 4-(2-chloroethyl)morpholine hydrochloride (70.8 mg, 0.381 mmol), K₂CO₃ (175 mg, 1.27 mmol), and KI (7.0 mg, 0.042 mmol) were added. The resulting mixture was heated to 90 °C for 2 hours. The reaction mixture was purified by preparative HPLC (method: Waters 2767 / 2545 / 2489, Inertsil ODS-3 10 μm 20*250 nm, mobile phase A: water containing 0.1% FA, mobile phase B: CH₃CN, flow rate: 20 mL / min, column temperature: RT) to give the title compound (13 mg, yield: 6.3%) as a white solid. LC-MS(ESI): m / z 445.1 [M+H] + . 1 HNMR (400MHz, DMSO-d6) δ8.76(t,J=5.5Hz,1H),7.93(d,J=4.3Hz,1H),7.83(d,J=8.6Hz,1H),7.77(d,J=2.8Hz,1H),7.26–7.30(m,1H),6 .63–6.70(m,2H),4.75(d,J=5.5Hz,2H),4.17(t,J=5.6Hz,2H),3.94(s,3H),3.55–3.61(m,4H),2.70(t,J=5.6Hz,2H),2.45–2.49(m,4H).

[0516] The following compounds were synthesized according to the procedure described in Example 38.

[0517]

[0518]

[0519] Example 39

[0520]

[0521] 2-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)-3,4-dihydroisoquinoline-1(2H)-one

[0522] a)N'-(2-chloroacetyl)thiophene-2-carbazide

[0523] A solution of 2-chloroacetyl chloride (4.20 mL, 52.8 mmol) in 10 mL THF was added dropwise to a mixture of thiophene-2-carbazide (5.00 g, 35 mmol), NaHCO3 (8.86 g, 106 mmol), H2O (40 mL), and THF (60 mL). The mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with ethyl acetate (50 mL) and washed with brine (10 mL). The organic layer was separated and concentrated under reduced pressure to give the title compound (2.10 g, yield: 27%). LC-MS (ESI): m / z 219.0 [M+H] + .

[0524] b) 2-(chloromethyl)-5-(thien-2-yl)-1,3,4-oxadiazole

[0525] A mixture of N'-(2-chloroacetyl)thiophene-2-carbonylhydrazine (1.00 g, 4.57 mmol) and POCl3 (8.50 mL, 91.5 mmol) was heated to 100 °C for 2 hours. The reaction mixture was then concentrated under reduced pressure. The residue was mixed with DCM (10 mL) and a saturated aqueous solution of NaHCO3 (20 mL). The organic layer was separated, dried over anhydrous Na2SO4, and filtered. The filtrate was concentrated under reduced pressure to give the title compound (700 mg, yield: 76%). LC-MS (ESI): m / z 201.0 [M+H] + .

[0526] c) 2-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl)-3,4-dihydroisoquinoline-1(2H)-one

[0527] NaH (74.8 mg, 1.87 mmol, 60% in oil) was added to a solution of 2-(chloromethyl)-5-(thiophen-2-yl)-1,3,4-oxadiazole (250 mg, 1.25 mmol) in DMF (15 mL) at 0 °C, and the resulting mixture was stirred at this temperature for 30 min. Then, 3,4-dihydroisoquinoline-1(2H)-one (250 mg, 1.25 mmol) was added. The reaction mixture was heated to room temperature and stirred for 1 hour. The resulting mixture was mixed with EA (100 mL) and brine (100 mL). The organic layer was separated, dried over anhydrous Na₂SO₄, and filtered under vacuum. The filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC (system: Waters 2767 / 2545 / 2489 / Qda, column: Inertsil ODS-3 10µm 20*250nm, mobile phase A: water containing 0.1% FA, mobile phase B: CH3CN, wavelength: 254nm / 214nm, flow rate: 20mL / min, column temperature: RT) to give the title compound (43.1mg, yield: 11%). LC-MS (ESI): m / z 312.0 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ7.94(d,J=4.9Hz,1H),7.77–7.85(m,2H),7.54–7.64(m,2 H),7.17–7.36(m,1H),5.02(s,2H),3.75(t,J=6.5Hz,2H),3.07(t,J=6.4Hz,2H).

[0528] Example 40

[0529]

[0530] 6-Bromo-2-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)-3,4-dihydroisoquinoline-1(2H)- ketone

[0531] NaH (178 mg, 4.49 mmol, 60% in oil) was added to a solution of 2-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)-3,4-dihydroisoquinoline-1(2H)-one (676 mg, 2.99 mmol) in DMF (15 mL) at 0 °C, and the reaction mixture was stirred at 0 °C for 30 min. Then, 6-bromo-3,4-dihydroisoquinoline-1(2H)-one (600 mg, 3.0 mmol) was added. The resulting mixture was heated to room temperature and stirred for 1 hour. The reaction mixture was diluted with ethyl acetate (20 mL) and brine (20 mL). The organic layer was separated, dried over anhydrous Na₂SO₄, and filtered. The filtrate was concentrated under reduced pressure to give the crude product (400 mg). The crude product (100 mg) was purified by preparative HPLC (method: Waters 2767 / 2545 / 2489 / Qda, column: Inertsil ODS-310um 20*250nm, mobile phase A: water containing 0.1% FA, mobile phase B: CH3CN, flow rate: 20 mL / min, column temperature: room temperature) to give the title compound (25 mg, yield: 6.5%) as a white solid. LC-MS (ESI): m / z 392.2 [M+H] + . 1 H NMR(400MHz,DMSO-d6)δ7.94(d,J=4.9Hz,1H),7.77-7.85(m,2H),7.54–7.64(m,2 H),7.17–7.36(m,1H),5.02(s,2H),3.75(t,J=6.5Hz,2H),3.07(t,J=6.4Hz,2H).

[0532] Example 41

[0533]

[0534] 6-Morphyrin-2-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)-3,4-dihydroisoquinoline-1 (2H)-ketone

[0535] A mixture of 6-bromo-2-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)-3,4-dihydroisoquinoline-1(2H)-one (300 mg, 0.77 mmol), morpholine (0.14 mL, 1.54 mmol), Cs₂CO₃ (749 mg, 2.30 mmol), BINAP (47.9 mg, 0.08 mmol), Pd₂(dba)₃ (70.3 mg, 0.080 mmol), and toluene (10 mL) was degassed three times with argon, then heated to 115 °C for 36 hours. The resulting mixture was filtered, and the filtrate was concentrated under vacuum. The residue was purified by preparative HPLC (system: Waters 2767 / 2545 / 2489 / Qda, Waters Xbridge C18 10µm OBD 19*150mm, mobile phase A: water containing 0.1% NH4OH, mobile phase B: CH3CN, flow rate: 20mL / min, column temperature: room temperature) to give the title compound (17.0 mg, yield: 5.6%) as a white solid. LC-MS (ESI): m / z 397.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ7.94(d,J=4.5Hz,1H),7.79(d,J=2.9Hz,1H),7.73(d,J=8.8Hz,1H),7.24–7.32 (m,1H),6.90(d,J=7.2Hz,1H),6.80(s,1H),4.97(s,1H),3.61–3.80(m,6H),3.24(s,2H),2.97(s,1H).

[0536] Example 42

[0537]

[0538] N-(2-Methoxy-4-morpholinylphenyl)-5-(thien-2-yl)-1,3,4-oxadiazol-2-carboxamide

[0539] a) 4-(3-methoxy-4-nitrophenyl)morpholine

[0540] Morpholine (4.07 g, 467 mmol) and potassium carbonate (7 mg, 51 mmol) were added to a solution of 4-fluoro-2-methoxy-1-nitrobenzene (8.00 g, 47 mmol) in acetonitrile (100 mL), and the reaction mixture was stirred at 75 °C for 3 h. The reaction mixture was concentrated under reduced pressure. The residue was mixed with water (30 mL), and the suspension was filtered. The filter cake was collected, washed with water (30 mL), and dried under vacuum to give the title compound (10.0 g, yield: 90%). LC-MS (ESI): m / z 239.1 [M+H] + .

[0541] b) 2-Methoxy-4-morpholinoaniline

[0542] Pd / C (10%, 500 mg) was added to a solution of 4-(3-methoxy-4-nitrophenyl)morpholine (3.00 g, 12.6 mmol) in ethyl acetate (30 mL), and the reaction mixture was degassed three times with N2. The reaction mixture was stirred at room temperature under H2 balloon pressure for 3 hours. The resulting mixture was filtered, and the filtrate was concentrated under reduced pressure to give the title compound (2.30 g, yield: 88%). LC-MS (ESI): m / z 209.0 [M+H] + .

[0543] c) Methyl 2-oxo-2-(2-(thiophene-2-carbonyl)hydrazino)acetate

[0544] A solution of methyl 2-chloro-2-oxoacetate (3.9 mg, 32 mmol) in 2.0 mL of THF was added dropwise to a mixture of thiophene-2-carbonylhydrazine (3.00 g, 21.1 mmol), NaHCO3 (5.32 g, 63.3 mmol), H2O (15 mL), and THF (15 mL). The reaction mixture was stirred at room temperature for 2 hours. The resulting mixture was concentrated under reduced pressure to remove most of the THF. The residue was extracted with EA (20 mL × 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous Na2SO4, and concentrated under reduced pressure to give the title compound (5.30 g, 100% yield). LC-MS (ESI): m / z 229.1 [M+H] + .

[0545] d) Methyl 5-(thien-2-yl)-1,3,4-oxadiazole-2-carboxylate

[0546] TEA (2.56 mL, 18.4 mmol) and toluenesulfonyl chloride (1.40 g, 7.36 mmol) were added to a solution of methyl 2-oxo-2-(2-(thiophene-2-carbonyl)hydrazido)acetate (1.4 g, 6.13 mmol) in DCM (20 mL). The resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with petroleum ether containing ethyl acetate (PE / EA = 1 / 10 to 1 / 1), to give the title compound (650 mg, yield: 50%). LC-MS (ESI): m / z 221.0 [M+H] + .

[0547] e)N-(2-methoxy-4-morpholinylphenyl)-5-(thien-2-yl)-1,3,4-oxadiazol-2-carboxamide

[0548] 2-Methoxy-4-morpholinoaniline (772 mg, 3.71 mmol) and TEA (0.86 mL, 6.18 mmol) were added to a solution of methyl 5-(thien-2-yl)-1,3,4-oxadiazol-2-carboxylate (130 mg, 0.62 mmol) in methanol (5 mL), and the reaction mixture was stirred at 65 °C for 5 h. The reaction mixture was concentrated under reduced pressure. The residue was mixed with ethanol (10 mL) and filtered under vacuum. The filter cake was dried under vacuum to give the title compound (27 mg, yield: 11%). LC-MS (ESI): m / z 387.0 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ9.87(s,1H),8.05(d,J=4.8Hz,1H),7.97(d,J=2.9Hz,1H),7.66(d,J=8.7Hz ,1H),7.44–7.27(m,1H),6.70(s,1H),6.55(d,J=8.5Hz,1H),3.87(s,3H),3.75(s,4H),3.15(s,4H).

[0549] Example 43

[0550]

[0551] 2-Methoxy-4-((1-methylpyrrolidone-3-yl)amino)-N-((5-(thiophene-2-yl)-1,3,4-oxadiazole- 2-yl)methyl)benzamide

[0552] a) 3-((3-methoxy-4-(methoxycarbonyl)phenyl)amino)pyrrolidine-1-carboxylic acid tert-butyl ester

[0553] Pd₂(dba)₃ (1.87 g, 2.04 mmol), BINAP (1.27 g, 2.04 mmol), tert-butyl 3-aminopyrrolidine-1-carboxylate (4.18 g, 22.44 mmol), and Cs₂CO₃ (20 g, 61 mmol) were added to a solution of methyl 4-bromo-2-methoxybenzoate (5.0 g, 20 mmol) in 1,4-dioxane (50 mL), and the reaction mixture was stirred overnight at 105 °C. The mixture was cooled to room temperature and diluted with ethyl acetate (200 mL) and brine (200 mL). The organic layer was separated and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with petroleum ether containing ethyl acetate (EA / PE = 1 / 100 to 1 / 2), to give the title compound (7.0 g, yield: 93%) as a yellow oil. LC-MS (ESI): m / z 351.5 [M+H] + .

[0554] b) Methyl 2-methoxy-4-(pyrrolidone-3-ylamino)benzoate hydrochloride

[0555] A mixture of tert-butyl 3-((3-methoxy-4-(methoxycarbonyl)phenyl)amino)pyrrolidine-1-carboxylate (7.0 g, 20 mmol) in HCl (4 M in dioxane, 10 mL) was stirred at room temperature for 18 hours. The mixture was concentrated under vacuum at 50 °C and dried to give the title compound (5.3 g, yield: 92.5%) as a yellow solid. LC-MS (ESI): m / z 251.1 [M-HCl+H] + .

[0556] c) Methyl 2-methoxy-4-((1-methylpyrrolidin-3-yl)amino)benzoate

[0557] Formaldehyde (0.21 g, 6.97 mmol, 30% in water) and sodium cyanoborohydride (0.26 g, 4.18 mmol) were added to a solution of methyl 2-methoxy-4-(pyrrolidone-3-ylamino)benzoate hydrochloride (1.0 g, 3.49 mmol) in methanol (20 mL), and the reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under vacuum, and the residue was purified by silica gel column chromatography by elution with dichloromethane containing methanol (MeOH / DCM = 0 / 1 to 1 / 15, v / v) to give the title compound (430 mg, yield: 46.8%) as a yellow solid. LC-MS (ESI): m / z 264.9 [M+H] + .

[0558] d) 2-Methoxy-4-((1-methylpyrrolidone-3-yl)amino)benzoic acid

[0559] A mixture of methyl 2-methoxy-4-((1-methylpyrrolidone-3-yl)amino)benzoate (600.00 mg, 1.63 mmol), LiOH (684 mg, 16.28 mmol), MeOH (20 mL), and H₂O (10 mL) was heated to 70 °C and stirred at this temperature for 18 hours. The reaction mixture was cooled to room temperature. The pH of the mixture was adjusted to 6–7. The mixture was concentrated under vacuum to obtain a residue. EtOH (50 mL) was added, and the mixture was stirred at room temperature for 20 minutes and filtered. The filtrate was concentrated under vacuum to give the title compound (400 mg, yield: 88%) as a white foam. LC-MS (ESI): m / z 250.9 [M+H] + .

[0560] e)2-Methoxy-4-((1-methylpyrrolidin-3-yl)amino)-N-((5-(thiophen-2-yl)-1,3,4-oxadiazine) (2-yl)methyl)benzamide

[0561] DIEA (524 mg, 4.06 mmol) and BOP (539 mg, 1.22 mmol) were added to a solution of 2-methoxy-4-((1-methylpyrrolidin-3-yl)amino)benzoic acid (400 mg, 1.60 mmol) in DMF (20 mL), and the reaction mixture was stirred at room temperature for 5 min. DMF (20 mL) containing (5-(thiophene-2-yl)-1,3,4-oxadiazol-2-yl)methylamine TFA salt (300 mg, 1.02 mmol) was added in a single-component manner. The reaction mixture was stirred at room temperature for 30 min. The mixture was purified by silica gel column chromatography by elution with methanol-containing DCM (MeOH / DCM = 0 / 1 to 1 / 10, v / v), and further purified by preparative HPLC (basic method) to give the title compound (0.16 g, yield: 38%) as a white solid. LC-MS(ESI): m / z 414.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.55(t,J=5.6Hz,1H),7.93(dd,J=5.0,1.1Hz,1H),7.77(dd,J=3.7,1. 1Hz,1H),7.67(d,J=8.4Hz,1H),7.27(dd,J=5.0,3.8Hz,1H),6.50(d,J=6.9Hz,1H),6.21(d,J= 8.5Hz,2H),4.73(d,J=5.6Hz,2H),3.96(s,1H),3.88(s,3H),2.73(dd,J=9.2,6.7Hz,1H),2.58 (dd,J=13.8,8.2Hz,1H),2.37(dt,J=9.7,6.0Hz,2H),2.29–2.19(m,4H),1.59(d,J=6.5Hz,1H).

[0562] Example 44

[0563]

[0564] 2-Methoxy-4-((2-(methylamino)ethyl)amino)-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2- α)(2-((3-methoxy-4-(((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)amino tert-butyl carbamate (carbamate)

[0565] Add (2-aminoethyl)(methyl)carbamate tert-butyl ester (199 mg, 1.1 mmol), Pd2(dba)3 (69.68 mg, 0.076 mmol), BINAP (47.38 mg, 0.076 mmol), and Cs2CO3 (991 mg, 3.04 mmol) to a solution of 4-bromo-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (300 mg, 0.761 mmol), to dioxane (6 mL), and degas the mixture three times with N2. Stir the reaction mixture at 105 °C for 18 hours. Filter the reaction mixture, dilute the filtrate with water (15 mL), and extract the mixture with ethyl acetate (20 mL × 3). Combine the organic layers, wash with brine (20 mL), dry to anhydrous Na2SO4, filter, and concentrate under vacuum. The residue was purified by silica gel column chromatography, eluting with petroleum ether containing ethyl acetate (PE:EA = 4:1 to 1:4) to give the title compound (200 mg, yield: 27%). LC-MS (ESI): m / z 488.2 [M+H] + .

[0566] b) 2-Methoxy-4-((2-(methylamino)ethyl)amino)-N-((5-(thien-2-yl)-1,3,4-oxadiazole- 2-yl)methyl)benzamide

[0567] TFA (2 mL) was added to a solution of (2-((3-methoxy-4-(((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)carbamoyl)phenyl)amino)ethyl)(methyl)carbamate tert-butyl ester (200 mg, 0.41 mmol) in dichloromethane (8 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was concentrated under vacuum. The residue was purified by preparative HPLC to give the title compound (17 mg, yield: 21%). LC-MS (ESI): m / z 388.2 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.54(d,J=5.4Hz,1H),8.01–7.87(m,1H),7.77(d,J=2.6Hz,1H),7.67(d,J=9.2Hz,1H),7.27(dd,J=4.9,3.8 Hz, 1H), 6.25 (t, J = 6.1Hz, 3H), 4.73 (d, J = 5.7Hz, 2H), 3.88 (s, 3H), 3.16 (dd, J = 11.8, 6.0Hz, 3H), 2.68 (t, J = 6.2Hz, 2H), 2.31 (s, 3H).

[0568] The following compounds were synthesized according to the procedure described in Example 44.

[0569]

[0570]

[0571] Example 45

[0572]

[0573] 2-Methoxy-4-(2-(methylamino)ethoxy)-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl) Methylbenzamide

[0574] a) 2-((tert-butoxycarbonyl)(methyl)amino)ethyl methanesulfonate

[0575] Triethylamine (693 mg, 6.8 mmol) and methanesulfonyl chloride (719 mg, 6.3 mmol) in DCM (10 mL) were added dropwise to a solution of tert-butyl (2-hydroxyethyl)(methyl)carbamate (1.0 g, 5.7 mmol) in DCM (10 mL) at 0 °C, and the reaction mixture was heated to room temperature. The reaction mixture was stirred at this temperature for 1.5 h. The reaction mixture was washed with water (10 mL), and the organic layer was separated. The organic layer was concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with petroleum ether containing ethyl acetate (PE:EA = 10:1 to 1:1), to give the title compound (1.16 g, yield: 80%) as a yellow oil.

[0576] b)(2-(3-methoxy-4-(((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl)carbamoyl) phenoxyethyl(methyl)carbamate tert-butyl

[0577] To a solution of 2-((tert-butoxycarbonyl)(methyl)amino)ethyl methanesulfonate (458 mg, 1.81 mmol) in DMF (5 mL), 4-hydroxy-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (300 mg, 0.91 mmol) and Cs₂CO₃ (589 mg, 1.81 mmol) were added, and the reaction mixture was stirred at 90 °C for 5 hours. The reaction mixture was diluted with water (20 mL), and the mixture was extracted with ethyl acetate (20 mL × 3). The organic layers were combined, washed with brine (20 mL), dried over anhydrous Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with petroleum ether containing ethyl acetate (PE:EA = 10:1 to 1:4), to give the title compound (53 mg, yield: 6.0%). LC-MS(ESI): m / z 489.0 [M+H] + .

[0578] c) 2-Methoxy-4-(2-(methylamino)ethoxy)-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2- methylbenzamide

[0579] TFA (1 mL) was added to a solution of (2-(3-methoxy-4-(((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)carbamoyl)phenoxy)ethyl)(methyl)carbamate (53 mg, 0.11 mmol) in DCM (5 mL), and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was concentrated under vacuum. The residue was purified by preparative HPLC to give the title compound (9.94 mg, yield: 24%). LC-MS (ESI): m / z 389.1 [MH] - . 1 H NMR (400MHz, DMSO-d6) δ8.76(t,J=5.5Hz,1H),7.93(d,J=4.9Hz,1H),7.84(d,J=8.7Hz,1H),7.77(d,J=3.3Hz,1H),7.32–7.18(m,1H) ,6.74–6.57(m,2H),4.75(d,J=5.6Hz,2H),4.10(t,J=5.5Hz,2H),3.93(s,3H),3.32–3.28(m,1H),2.87(t,J=5.3Hz,2H),2.35(s,3H).

[0580] The following compounds were synthesized according to the procedure described in Example 45.

[0581]

[0582] Example 46

[0583]

[0584] 2-Methoxy-4-((1-methylpyrrolidone-3-yl)oxy)-N-((5-(thiophene-2-yl)-1,3,4-oxadiazole- 2-yl)methyl)benzamide

[0585] Acetic acid (22 mg, 0.12 mmol), formaldehyde (126 mg, 4.20 mmol), and 2-methylpyridineborane (641 mg, 6.0 mmol) were added to a solution of 2-methoxy-4-(pyrrolidine-3-yloxy)-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (240 mg, 0.6 mmol) in methanol (10 mL), and the reaction mixture was stirred at 80 °C for 2 hours. The reaction mixture was concentrated under vacuum. The residue was purified by silica gel column chromatography by elution with methanol-containing DCM (MeOH:DCM = 1:100 to 1:10, v / v) to give the crude product. The crude product was further purified by preparative HPLC to give the title compound (2.18 mg, yield: 0.8%) as a white solid. LC-MS (ESI): m / z 415.5 [M+H] + .1 H NMR (400MHz, DMSO-d6) δ8.75(t,J=5.4Hz,1H),7.93(dd,J=5.0,1.1Hz,1H),7.81(s,1H),7.77(dd,J=3.7,1.1Hz,2H),7.28(dd ,J=5.0,3.7Hz,1H),6.60(m,2H),4.97(s,1H),4.75(d,J=5.6Hz,2H),3.92(s,3H),2.86-2.73(m,1H),2.72-2.60(m,2H),2.41 -2.31(m,2H),2.27(s,3H),1.76(d,J=7.1Hz,1H).

[0586] Example 47

[0587]

[0588] 4-(2-hydroxypropyl-2-yl)-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl) benzamide

[0589] a) 4-(2-hydroxypropyl-2-yl)-2-methoxybenzoic acid

[0590] A solution of 4-bromo-2-methoxybenzoic acid (400 mg, 1.73 mmol) in THF (5 mL) was cooled to -65 °C, and n-BuLi (3.3 mL, 0.48 mmol, 1.6 M) was added. The reaction mixture was stirred at this temperature for 30 min. Acetone (0.19 mL, 2.5 mmol) was added dropwise to the mixture, and the mixture was slowly warmed to 0 °C and stirred for 30 min. The mixture was diluted with ethyl acetate (5 mL) and saturated NH4Cl aqueous solution (5 mL). The organic layer was separated, washed with brine (5 mL), filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with dichloromethane containing methanol (MeOH:DCM = 0:1 to 1:7), to give the title compound (50 mg, yield: 14%) as a colorless oil. LC-MS (ESI): m / z 211.0 [M+H] + .

[0591] b) 4-(2-hydroxypropyl-2-yl)-2-methoxy-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl (Bento) benzamide

[0592] BOP (210 mg, 0.48 mmol), DIEA (0.12 mL, 0.71 mmol), and [5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl]methylamine (43 mg, 0.24 mmol) were added to a solution of 4-(2-hydroxypropyl-2-yl)-2-methoxybenzoic acid (50 mg, 0.24 mmol) in DMF (5 mL). The reaction mixture was stirred at room temperature for 1.5 h. The reaction mixture was purified by preparative HPLC (Waters 2767 / 2545 / 2489, Waters Xbridge C18 10 μm OBD 19*250 mm, mobile phase A: water containing 0.1% NH4HCO3, mobile phase B: CH3CN, 20 mL / min, RT) to give the title compound (7 mg, yield: 7.6%) as a white solid. LC-MS(ESI): m / z 374.1 [M+H] + . 1 H NMR (400MHz, DMSO-d6) δ8.88(t,J=5.7Hz,1H),7.94(d,J=4.0Hz,1H),7.76(t,J=6.1Hz,2H),7.27(dd,J =9.5, 4.6Hz, 2H), 7.11 (d, J = 8.1Hz, 1H), 5.18 (s, 1H), 4.77 (d, J = 5.6Hz, 2H), 3.93 (s, 3H), 1.44 (s, 6H).

[0593] Example 48

[0594]

[0595] 4-(2-hydroxy-2-methylpropyl)-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl (Bento) benzamide

[0596] a) Methyl 4-(2-hydroxy-2-methylpropyl)-2-methoxybenzoate

[0597] LDA (8.3 mL, 0.2 mmol, 2 M in THF) was added at -70 °C to a solution of 1,3-dimethyl-1,3-diazin-2-one (7.8 mL, 64 mmol) in 30 mL of THF, followed by the addition of methyl 2-methoxy-4-methylbenzoate (2.91 g, 16 mmol). The reaction mixture was stirred at -70 °C for 2 hours, and acetone (1.78 mL, 24.224 mmol) was added. The reaction mixture was heated to 0 °C and stirred at that temperature for 1 hour. The mixture was diluted with brine (20 mL) and extracted with ethyl acetate (40 mL × 2). The combined organic layers were washed with brine (20 mL), dried over Na₂SO₄, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with petroleum ether containing ethyl acetate (PE / EA = 1 / 0 to 1.5 / 1) to give the title compound (900 mg, yield: 19.2%) as a yellow oil. LC-MS (ESI): m / z 238.8 [M+H] + .

[0598] b) 4-(2-hydroxy-2-methylpropyl)-2-methoxybenzoic acid

[0599] H₂O (20 mL) and LiOH·H₂O (3.00 g, 71 mmol) were added to a solution of methyl 4-(2-hydroxy-2-methylpropyl)-2-methoxybenzoate (1.7 g, 7.13 mmol) in MeOH (20 mL). The reaction mixture was stirred at 60 °C for 2 h. The mixture was cooled to room temperature and the pH was adjusted to 7–8 with an aqueous HCl solution (1.0 M). The organic solvent was removed under vacuum, and water (20 mL) was added. The mixture was acidified with hydrochloric acid (5 mL) and extracted with ethyl acetate (3 × 50 mL). The combined organic layers were dried over Na₂SO₄, filtered, and concentrated to give the title compound (1.4 g, yield: 72%) as a yellow oil. LC-MS (ESI): m / z 225.1 [M+H] + .

[0600] c) 4-(2-hydroxy-2-methylpropyl)-2-methoxy-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl) Methylbenzamide

[0601] To a solution of (5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methylamine TFA salt (2.19 g, 5.563 mmol) in DMF (15 mL), DIEA (2.51 mL, 15.171 mmol), BOP (2.68 g, 6.068 mmol), and 4-(2-hydroxy-2-methylpropyl)-2-methoxybenzoic acid (1.4 g, 5.057 mmol) were added. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was diluted with ethyl acetate (30 mL) and brine (50 mL). The organic layer was separated, dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography by elution with petroleum ether containing ethyl acetate (PE:EA = 1:0–1 / 2) to give the title compound (1.0 g, 2.59 mmol, yield: 51%) as a grayish-white solid. LC-MS(ESI): m / z 388.1 [M+H] + . 1 HNMR (400MHz, DMSO-d6) δ8.87(t,J=5.6Hz,1H),7.93(dd,J=5.0,1.2Hz,1H),7.78(dd,J=3.7,1.2Hz,1H),7.73(d,J=7.9Hz,1H),7.28(dd ,J=5.0,3.7Hz,1H),7.02(s,1H),6.90(dd,J=7.9,1.1Hz,1H),4.77(d,J=5.7Hz,2H),4.39(s,1H),3.91(s,3H),2.70(s,2H),1.09(s,6H).

[0602] Example 49

[0603]

[0604] 2-Methoxy-4-(piperidin-1-ylmethyl)-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl) benzamide

[0605] a) 4-Formyl-2-methoxybenzoic acid

[0606] A solution of 4-bromo-2-methoxybenzoic acid (4.0 g, 17 mmol) in THF (50 mL) was cooled to -65 °C, and n-butyllithium (15.2 mL, 0.16 mmol, 2.5 M in THF) was added. The reaction mixture was stirred at this temperature for 30 min. DMF (1.62 mL, 21 mmol) was added dropwise to this mixture, and the mixture was then slowly warmed to 0 °C and stirred for 30 min. The mixture was diluted with ethyl acetate (30 mL), washed with saturated aqueous NH4Cl solution (50 mL) and brine (50 mL). The organic layer was dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with dichloromethane containing methanol (MeOH:DCM = 0:1 to 1:20), to give the title compound (2.3 g, yield: 55%) as a yellow solid. LC-MS (ESI): m / z 179.1 [MH] - .

[0607] b) 4-Formyl-2-methoxy-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide

[0608] BOP (828 mg, 1.9 mmol), DIEA (0.62 mL, 3.7 mmol), and (5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methylamine TFA salt (347.19 mg, 1.249 mmol) were added to a solution of 4-formyl-2-methoxybenzoic acid (300 mg, 1.249 mmol) in DMF (10 mL). The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was diluted with ethyl acetate (20 mL) and brine (40 mL). The organic layer was separated, washed with brine (40 mL), dried over Na2SO4, filtered, and concentrated under vacuum. The residue was purified by silica gel column chromatography, eluting with dichloromethane containing methanol (MeOH / DCM = 0 / 1 to 1 / 20), to give the title compound (120 mg, yield: 26%) as a white solid. LC-MS(ESI): m / z 344.1 [M+H] + .

[0609] c) 2-Methoxy-4-(piperidin-1-ylmethyl)-N-((5-(thien-2-yl)-1,3,4-oxadiazol-2-yl)methyl (Bento) benzamide

[0610] To a solution of 4-formyl-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (50 mg, 0.15 mmol) in MeOH (5 mL), piperidine (0.2 mL, 2.02 mmol) and acetic acid (0.01 mL, 0.175 mmol) were added, and the reaction mixture was stirred at room temperature for 1 hour. Sodium cyanoborohydride (18 mg, 0.291 mmol) was then added to this mixture, and the reaction mixture was stirred at the same temperature for 2 hours. After the reaction was complete, the mixture was purified by preparative HPLC (Waters 2767 / 2545 / 2489, Waters Xbridge C18 10um OBD 19*250mm, mobile phase A: water containing 0.1% NH4OH, mobile phase B: CH3CN, flow rate: 20 mL / min, column temperature: RT) to give the title compound (18 mg, yield: 30%) as a white solid. LC-MS (ESI): m / z 413.1 [M+H] + . 1 H NMR(400MHz, DMSO-d6)δ8.89(t,J=5.7Hz,1H),7.94(dd,J=5.0,1.2Hz,1H),7.79–7.75(m,2H),7.28(dd,J=5.0,3.7Hz,1H),7.0 8(s,1H),6.99(d,J=7.3Hz,1H),4.77(d,J=5.7Hz,2H),3.91(s,3H),3.46(s,2H),2.33(s,4H),1.53–1.47(m,4H),1.39(s,2H).

[0611] The following compounds were synthesized according to the procedure described in Example 49.

[0612]

[0613] Bioanalysis and Data

[0614] As described above, the compound of formula I is a MIF inhibitor and can be used to treat MIF-mediated diseases. The bioactivity of the compound can be determined by any analytical method suitable for assessing the activity of the candidate compound as a MIF inhibitor.

[0615] MIF enzyme analysis

[0616] Tautomerase analysis using pHPP as a substrate

[0617] This analysis determined the activity of the MIF tautomerase by measuring the initial rate of the keto-enol tautomerization of pHPP catalyzed by MIF in a cell-free system. This was achieved by spectrophotometric quantification of the complex between the borate and the reaction product (enol pHPP). The substrate was prepared by converting enol pHPP to its ketone form. For this purpose, 0.5 M pHPP in methanol was diluted 10-fold with 50 mM sodium acetate buffer at pH 6.0, and the suspension was then agitated in the dark at room temperature for 24 hours. Finally, it was stored at 4°C for no more than one week, and it is recommended to sonicate for 5 minutes before use.

[0618] Analysis was performed in small-volume black-bottomed 96- or 384-well polystyrene plates (Greiner Bio-One). First, 2 μL of D-PBS enzyme solution containing 6 nM MIF (with 0.025% w / v BSA and 300 μM CHAPS) was aliquoted into the sample and negative control wells using a Multidrop Combi (Thermo Fisher Scientific) dispenser with a metal capillary box, pre-treated with Sigmacote silicate. Then, 2 μL of the same buffer solution without MIF was aliquoted into the positive control wells. The reaction was initiated by adding the following to all wells: 2 μL of substrate solution containing 3 mM ketone pHPP in 200 mM borate, 25 mM sodium phosphate, 0.025% w / v BSA, and 300 μM CHAPS, pH 6.0. The plates were centrifuged at 1000 rpm for 2 minutes at room temperature in an Allegra 25R centrifuge (Beckman Coulter, Inc., Brea, CA) to remove air bubbles. Then, the plate was read using EnVision. The final concentrations of the enzyme and substrate were 3 nM and 1.5 mM, respectively. The initial rate for each well was calculated as the slope of the absorbance progression curve.

[0619] All example compounds (Examples 1-85) were tested in the above-described MIF tautomerase assays or similar assays. The data mentioned below represent the average pIC values ​​from multiple test results. 50 Values. It is understandable that the data shown below may vary reasonably depending on the specific conditions and procedures used by the personnel conducting the tests.

[0620] Analyze data

[0621] Analytical results of pIC measurement of the inhibitory effect of the compound on MIF 50 The data is listed in Table 1 below.

[0622] Table 1. In vitro inhibition of MIF enzyme by the compounds

[0623]

[0624]

[0625] Cell proliferation analysis

[0626] This analysis used the Cell Counting Kit-8 (CCK-8) to detect the ability of the compound to inhibit cell proliferation. This kit uses sensitive colorimetric analysis to determine cell viability in cell proliferation and cytotoxicity assays. The highly water-soluble tetrazolium salt WST-8 is reduced by dehydrogenase activity in cells to produce a yellow formazan dye, soluble in tissue culture medium. The amount of formazan dye produced by dehydrogenase activity in cells is directly proportional to the number of viable cells.

[0627] Analysis was performed according to the manufacturer's (Dojindo Molecular Technologies Inc., Rockville, MD, USA) instructions. To begin the analysis, BV2 cells were loaded at 2 × 10⁻⁶ cells / mL. 6 Cells were seeded at a density of 10 cells / well in 96-well plates and treated with the assay compound, with a maximum final analytical concentration of 33.3 μM. The assay compound was diluted 1:3 with analytical medium to generate 10 concentration responses over 48 hours. After incubation, 10 μl of CCK-8 solution was added to each well of the cell culture plate, and the plate was incubated at 37°C for 3 hours. Absorbance was independently recorded three times at 450 nm using a microplate reader (Bio-Rad Laboratories, Richmond, CA, USA).

[0628] The selected examples were tested in BV2 cell proliferation. The data mentioned below represent the average pIC of multiple test results. 50 Values. It is understandable that the data shown below may vary reasonably depending on the specific conditions and procedures used by the personnel conducting the tests.

[0629] Analytical results of pIC measurement of the inhibitory effect of the compound on MIF 50 The data is listed in Table 2 below.

[0630] Table 2. Inhibition of BV2 and U251 cell proliferation by the compounds

[0631]

[0632]

Claims

1. A compound or a pharmaceutically acceptable salt thereof, wherein the compound is selected from... 4-(benzoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (compound 15); 4-Hydroxy-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (compound 16); 4-(2-hydroxyethoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (compound 17); 4-(2-(benzoxy)ethoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (compound 18); 2-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenoxy)acetic acid (compound 19); 4-(2-(dimethylamino)ethoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (compound 20); 2-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenoxy)-2-methylpropionic acid (compound 21); 4-((2H-tetrazol-5-yl)methoxy)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (compound 22); 2-Methoxy-4-((2-((5-methyl-2-oxo-1,3-dioxacyclopenten-4-yl)methyl)-2H-tetrazole-5-yl)methoxy)- N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (compound 23); 2-Methoxy-4-((1-((5-methyl-2-oxo-1,3-dioxacyclopenten-4-yl)methyl)-1H-tetrazole-5-yl)methoxy)- N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (compound 24); 4-(3-cyanopropyl)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (compound 25); 4-(3-(2H-tetrazol-5-yl)propyl)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (compound 26); cis-4-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenyl)cyclohexane-1-carboxylic acid (compound 27); trans-4-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenyl)cyclohexane-1-carboxylic acid (compound 28); 3'-Methoxy-4'-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)-2,3,4,5-tetrahydro-[1,1'-biphenyl]-4-carboxylic acid (compound 29); (3-Methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenyl)proline (compound 30); 1-(3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenyl)piperidine-3-carboxylic acid (compound 31); 2-((3-methoxy-4-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)carbamoyl)phenyl)amino)-2-methylpropionic acid (compound 32); 4-(6-oxa-3-azabicyclo[3.1.1]hept-3-yl)-2-methoxy-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (compound 33); 2-Methoxy-4-morpholino-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (compound 37); 2,4-Dimethoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 38); 2-Methoxy-4-morpholino-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 51); 4-(2,6-dimethylmorpholino)-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 55); 2-Methoxy-4-(piperidin-1-yl)-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 56); 4-(6-oxa-3-azabicyclo[3.1.1]hept-3-yl)-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 57); 2-Methoxy-4-(piperazin-1-yl)-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 58); 2-Methoxy-4-(4-methylpiperazin-1-yl)-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 59); 2-Methoxy-4-(2-morpholinylethoxy)-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 69); 4-(2-hydroxyethoxy)-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 70); 2-Methoxy-4-(2-Methoxyethoxy)-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 71); 4-(3-hydroxypyrrolidone-1-yl)-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 78); 4-((2-hydroxyethyl)amino)-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 79); or 4-(2-hydroxypropyl-2-yl)-2-methoxy-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 83).

2. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is 2-methoxy-4-morpholino-N-(5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)benzamide (compound 37).

3. The compound of claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound is 2-methoxy-4-morpholino-N-((5-(thiophen-2-yl)-1,3,4-oxadiazol-2-yl)methyl)benzamide (compound 51).

4. A pharmaceutical composition comprising a compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier or excipient.

5. Use of a compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof for the preparation of a medicament for treating diseases mediated by MIF.

6. The use according to claim 5, wherein the MIF-mediated disease is Tumors selected from glioblastoma, lung cancer, breast cancer, gastric cancer, bladder cancer, or melanoma; Inflammatory diseases selected from chronic obstructive pulmonary disease (COPD) or pneumonia; Autoimmune diseases selected from rheumatoid arthritis (RA), multiple sclerosis (MS), or systemic lupus erythematosus (SLE).

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