Heterobicyclic compounds as EP4 receptor antagonists

Abstract

The present invention relates to a series of novel heterobicyclic amide derivatives as EP4 receptor antagonists, which are useful for treating diseases or conditions mediated by the action of PGE2 at the EP4 receptor, such as pain, inflammatory diseases and cancer.The scope of the present invention also includes pharmaceutical compositions containing such compounds and methods for using these compounds to treat subjects suffering from conditions mediated by the action of PGE2 at the EP4 receptor.

Description

[Background technology]

[0001] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority to International Application No. PCT / CN2022 / 114402 (filed August 24, 2022) and U.S. Application No. US 63 / 425,555 (filed November 15, 2022), the contents of both of which are incorporated herein by reference in their entireties.

[0002] BACKGROUND OF THE INVENTION The present invention relates to heterocyclic amide derivatives, or pharmaceutically acceptable salts thereof, pharmaceutically acceptable prodrugs thereof, pharmaceutical compositions prepared therefrom, and their medical uses in mammals, including humans. The compounds of the present invention have activity as prostaglandin E2 (PGE2) receptor antagonists, which are useful in the treatment or alleviation of pain, inflammation, and cancer.

[0003] Prostaglandins are a group of bioactive lipid compounds with diverse hormone-like effects in the body. They are intriguing because they may exert different effects depending on the receptor to which they bind. Research has shown that prostaglandins are important mediators of pain, fever, and other inflammation-related symptoms. Of particular interest is prostaglandin E2 (PGE2), the major eicosanoid associated with inflammation. Furthermore, high expression of PGE2 in tumor tissue suppresses antitumor immunity in the tumor microenvironment (TME), causing tumor immune evasion and leading to disease progression (see Front Immunol. 2020; 11: 324). PGE2 has also been shown to promote the proliferation and metastasis of colorectal cancer stem cells (CSCs) in mice (Wang et al., Gastroenterology, 2015, 1-12). PGE2 activity is primarily mediated by its binding to a panel of G protein-coupled receptors (GPCRs), namely, the E-type prostanoid (EP) receptors EP1, EP2, EP3, and EP4. Among these, the EP4 receptor is currently considered the most versatile and promising of the PGE2 receptors. Animal studies have shown that selective inhibition of PGE2 / EP4 receptor signaling by antagonists suppresses tumor growth (Terada et al. Cancer Res. 2010, 70, 1606-1615) and tumor metastasis (Yang et al. Cancer Res. 2006, 66, 9665-9672).

[0004] Up to now, various structural classes of EP4 receptor antagonists have been reported.The object of the present invention is to provide a series of novel heterobicyclic compounds as EP4 receptor antagonists, and the method for treating diseases or symptoms including pain, inflammation and cancer mediated by the action of PGE2 at EP4 receptor, and their pharmaceutical compositions.

[0005] (Summary of the Invention) The present invention relates to a series of novel heterobicyclic amide derivatives as EP4 receptor antagonists, which are useful for treating diseases or conditions mediated by the action of PGE2 at the EP4 receptor, such as pain, inflammation, and cancer. Pharmaceutical compositions and methods of use are also included.

[0006] In one aspect, the present invention provides a heterobicyclic compound or a pharmaceutically acceptable salt thereof, the compound having the structure of Formula I, as shown below: [ka] During the ceremony: R 1 and R 2 are independently hydrogen, C 1-6 Alkyl, C 1-6 Cycloalkyl, C 1-6 Halocycloalkyl, and C 1-6 haloalkyl; or R 1 and R 2 together with the carbon atoms to which they are both attached form a 3- to 6-membered carbocyclic ring, and the carbocyclic ring is a groups, each independently S, O, or NR b and each Rb is independently selected from hydrogen, C 1-6 Alkyl, C 1-6 Cycloalkyl, C 1-6 Halocycloalkyl, and C 1-6 Haloalkyl, aryl, heteroaryl, -C(O)-C 1-6 alkyl, -C(O)-aryl, -S(O)2-alkyl, or -S(O)2-aryl; X is absent, =CH-, -CR 1 R 2 - or -C(O)-; Cy 1 But C 1-6 Alkylene, C 1-6 Alkenylene, C 1-6alkynylene, cycloalkylene, arylene, heteroarylene, heterocyclylene, or bridged bicyclic cycloalkylene, and is optionally substituted; Cy 2 is cycloalkyl, aryl, heteroaryl, or heterocyclyl, each of which is optionally substituted with 1 to 3 substituents that are each independently halo, alkyl, or haloalkyl; Each R a are independently halo, alkyl, haloalkyl, hydroxyalkyl, or alkoxy; R a If is alkyl, Cy 1 is a bridged bicyclic cycloalkylene.

[0007] In some embodiments, X is CH2.

[0008] In some embodiments, Cy 2 is aryl optionally substituted with one haloalkyl.

[0009] In some embodiments, halo is -F or -Cl.

[0010] In some embodiments, Cy 1 is an arylene or a bridged bicyclic cycloalkylene.

[0011] In some embodiments, R 1 and R 2 are each independently hydrogen or C alkyl; or R 1 and R 2 together with the carbon atoms to which they are both attached form a 3- to 6-membered carbocyclic ring.

[0012] In some embodiments, R a is -F, -Cl, -CF3, hydroxyalkyl, alkoxyl, or -CH3, with the proviso that R a If -CH3, Cy 1 C5-C10 It is a bridged bicyclic cycloalkylene.

[0013] In some embodiments, the compound is of Formula (II): [ka] where Cy 1 is arylene or bridged bicyclic cycloalkylene; R a is halo, alkyl, haloalkyl, hydroxyalkyl, or alkoxy, with the proviso that R a If is alkyl, Cy 1 is a bridged bicyclic cycloalkylene.

[0014] In yet another embodiment, Cy 1 but phenylene or C5-C 10 Bridged bicyclic cycloalkylene. C5-C 10 Examples of bridged bicyclic cycloalkylenes are: [ka] Including, but not limited to:

[0015] Examples of compounds of the present invention include: [ka]

[0016] Another aspect of the present invention provides pharmaceutical compositions comprising the described compounds or pharmaceutically acceptable salts thereof, and a pharmaceutically or physiologically acceptable carrier or excipient, respectively.

[0017] In some embodiments, such pharmaceutical compositions include another therapeutic agent, which may be an antibody against cytotoxic T-lymphocyte antigen 4 (anti-CTLA4), an antibody against programmed cell death ligand 1 (anti-PDL1), an antibody against programmed cell death protein 1 (anti-PD1), an indoleamine-2,3-dioxygenase (IDO) inhibitor, a tryptophan-2,3-dioxygenase (TDO) inhibitor, or an antimetabolite.

[0018] In some embodiments, such pharmaceutical compositions are used in combination with radiotherapeutic agents.

[0019] Yet another aspect of the present invention provides a method for treating a subject suffering from a condition mediated by the action of PGE2 at the EP4 receptor, comprising administering to a subject in need thereof an effective amount of the above-described compound or pharmaceutical composition.

[0020] In some embodiments, the condition is an inflammatory disease or cancer.

[0021] Examples of inflammatory diseases include, but are not limited to, arthritis, acne vulgaris, asthma, autoimmune diseases, autoinflammatory diseases, celiac disease, chronic prostatitis, colitis, diverticulitis, glomerulonephritis, hidradenitis suppurativa, hypersensitivity reactions, inflammatory bowel disease, interstitial cystitis, mast cell activation syndrome, macrocytosis, otitis, pelvic inflammatory disease, reperfusion injury, rheumatic fever, rheumatoid arthritis, rhinitis, sarcoidosis, or vasculitis.

[0022] Examples of cancer include, but are not limited to, breast cancer, endometrial cancer, cervical cancer, ovarian cancer, lung cancer, head and neck cancer, brain cancer, thyroid cancer, esophageal cancer, stomach cancer, colorectal cancer, liver cancer, pancreatic cancer, skin cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, bone cancer, lymphoma, and blood cancer.

[0023] Also included within the scope of the present invention is the use of the above compounds for the manufacture of a medicament for treating a subject suffering from a condition mediated by the action of PGE2 at the EP4 receptor. DETAILED DESCRIPTION OF THE INVENTION

[0024] (Detailed Description of the Invention) Reference will now be made in detail to preferred embodiments of the invention, examples of which will be further described. While the invention will be described in conjunction with preferred embodiments, it will be understood that it is not intended to limit the invention to these embodiments. On the contrary, the invention is intended to cover alternatives, modifications, and equivalents which may be included within the spirit and scope of the invention as defined by the claims. Furthermore, in the detailed description of the invention, numerous specific details are set forth in order to provide a thorough understanding of the invention. However, it will be apparent to those skilled in the art that the invention may be practiced without these specific details. In other instances, well-known methods, procedures, components, and other features have not been described in detail so as not to unnecessarily obscure aspects of the invention. definition

[0025] As used herein, the term "or" is meant to include both "and" and "or," i.e., the term "or" can also be replaced with "and / or."

[0026] As used herein, the singular forms "a," "an," and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise.

[0027] As used herein, the terms "subject" or "patient" are used interchangeably and, as used herein, refer to any mammal, including, but not limited to, a human patient or subject, to which the compositions of the present invention can be administered. The term "mammal" includes human patients, non-human primates, as well as laboratory animals such as rabbits, rats, and mice, and other animals.

[0028] As used herein, the term "unsaturated bond" refers to a double bond or a triple bond.

[0029] As used herein, the terms "unsaturated" or "partially unsaturated" refer to a moiety that contains at least one double or triple bond.

[0030] As used herein, the term "saturated" refers to a moiety that does not contain double or triple bonds, i.e., a moiety that contains only single bonds.

[0031] As used herein, the term "alkyl," by itself or as part of another substituent, refers to an alkyl group containing no unsaturation and having the specified number of carbon atoms (e.g., C-C 10 or C 1-10 "alkyl" refers to a straight-chain (i.e., unbranched) or branched hydrocarbon chain radical consisting of carbon and hydrogen atoms, having a number (e.g., alkyl) of 1 to 10 carbon atoms. Whenever a numerical range such as "1 to 10" is used herein, it refers to each integer in the specified range; for example, "1 to 10 carbon atoms" means that the alkyl group can be composed of 1 carbon atom, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, etc., up to 10 carbon atoms, although this definition also covers occurrences of the term "alkyl" when no numerical range is specified. Representative saturated straight-chain or linear alkyls include, but are not limited to, -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, and -n-hexyl; while saturated branched alkyls include, but are not limited to, -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, 2-methylbutyl, 3-methylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylbutyl, etc. The alkyl is connected to the parent molecule by a single bond. Unless otherwise specified herein, the alkyl group is optionally substituted with one or more substituents.

[0032] As used herein, the term "halo" or "halogen" refers to fluorine (fluoro, -F), chlorine (chloro, -Cl), bromine (bromo, -Br), or iodine (iodo, -I). "Haloalkyl" refers to an alkyl as defined above in which one or more hydrogen atoms have been replaced with a halogen independently selected from fluoro, chloro, bromo, and iodo. "Fluoroalkyl" refers to an alkyl as defined above in which one or more hydrogen atoms have been replaced with a fluoro atom.

[0033] As used herein, the term "alkenyl," by itself or as part of another substituent, refers to an unsaturated branched, straight-chain, or cyclic alkyl having at least one carbon-carbon double bond derived by the removal of a hydrogen atom from a single carbon atom of a parent alkene. The group can be in either the cis or trans conformation about the double bond(s). Typical alkenyl groups include, but are not limited to, ethenyl, propenyl, and the like.

[0034] The term "alkynyl," as used herein, by itself or as part of another substituent, refers to carbon chains which contain at least one carbon-carbon triple bond, and which may be linear or branched or combinations thereof. Examples of alkynyl include ethynyl, propargyl, 3-methyl-1-pentynyl, 2-heptynyl, and the like.

[0035] As used herein, the term "cycloalkyl," by itself or as part of another substituent, refers to a cyclic version of an "alkyl" group. A cycloalkyl group may contain zero bridgehead carbon atoms, or two or more. Thus, a cycloalkyl can be monocyclic, bicyclic, or polycyclic, depending on the number of bridgehead and bridging carbon atoms. A cycloalkyl group containing zero bridgehead carbon atoms is referred to herein as a "monocyclic cycloalkyl" or "unbridged cycloalkyl." A cycloalkyl containing at least two bridgehead carbon atoms and at least one bridging carbon atom is referred to herein as a "bridged cycloalkyl." A bridged cycloalkyl containing two bridgehead carbon atoms is referred to herein as a "bicyclic bridged cycloalkyl" or "bridged bicyclic cycloalkyl." A bridged cycloalkyl containing more than two bridgehead carbon atoms is referred to herein as a "polycyclic bridged cycloalkyl" or "bridged polycyclic cycloalkyl." A "lower" unbridged cycloalkyl contains 3 to 8 carbon atoms. A "lower" bridged cycloalkyl contains 5 to 16 carbon atoms.

[0036] As used herein, the term "cycloalkenyl" refers to a non-aromatic carbon-based ring composed of at least three carbon atoms and containing at least one carbon-carbon double bond, i.e., C=C. Examples of cycloalkenyl groups include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, and the like. The term "heterocycloalkenyl" refers to a type of cycloalkenyl group defined above, in which at least one of the carbon atoms of the ring is replaced with a heteroatom, such as nitrogen, oxygen, sulfur, or phosphorus, but is not limited to this. Cycloalkenyl and heterocycloalkenyl groups can be substituted or unsubstituted.

[0037] The term "heterocyclyl," as used herein, refers to a group derived from a monocyclic, bicyclic, or polycyclic compound containing at least one non-aromatic ring containing one or more, preferably one to three, heteroatoms independently selected from nitrogen, oxygen, and sulfur. The heterocyclyl groups of the present disclosure can be attached to the parent molecular moiety through a carbon atom or a heteroatom in the group.

[0038] As used herein, the term "hydroxyl" or "hydroxy" refers to an --OH group.

[0039] The groups defined above may contain prefixes and / or suffixes that are commonly used in the art and form further well-known substituents. As an example, the term "alkyloxy" or "alkoxy" refers to a group of the formula -OR, "alkylamine" refers to a group of the formula -NHR, and "dialkylamine" refers to a group of the formula -NRR, where each R is independently alkyl. As another example, "haloalkoxy" or "haloalkyloxy" refers to a group of the formula -OR', where R' is haloalkyl.

[0040] The term "hydroxyalkyl," by itself or as part of another substituent, refers to an alkyl group in which one or more hydrogen atoms have been replaced with a hydroxyl substituent. Thus, the term "hydroxyalkyl" is meant to include, for example, monohydroxyalkyl, dihydroxyalkyl, trihydroxyalkyl, etc.

[0041] The term "aryl," as used herein, refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 π electrons shared in a cyclic arrangement) having 6 to 14 ring carbon atoms and 0 heteroatoms arranged in the aromatic ring system ("C 6-14 In some embodiments, an aryl group has 6 ring carbon atoms ("C aryl", e.g., phenyl). In some embodiments, an aryl group has 10 ring carbon atoms ("C 10aryl," e.g., naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, the aryl group has 14 ring carbon atoms ("C 14 aryl", e.g., anthracyl).

[0042] As used herein, the term "heteroaryl" refers to a radical of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., having 6, 10, or 14 π electrons shared in a cyclic arrangement) having ring carbon atoms and one or more ring heteroatoms arranged in the aromatic ring system, where each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In heteroaryl groups containing one or more nitrogen atoms, the point of attachment can be a carbon atom or a nitrogen atom, as valence permits. Heteroaryl polycyclic ring systems can contain one or more heteroatoms in one or both rings.

[0043] The term "alkylene," by itself or as part of another substituent, refers to a bidentate moiety obtained by removing two hydrogen atoms, either both from the same carbon atom or one from each of two different carbon atoms, of a hydrocarbon compound that may be aliphatic or alicyclic, and that may be saturated, partially unsaturated, or fully unsaturated. Thus, the term "alkylene" includes the subclasses alkenylene, alkynylene, cycloalkylene, etc. In this context, prefixes (e.g., C 1-4 , C 1-7 , C 1-20 , C 2-7 , C 3-7 For example, the term "C1-4 alkylene" as used herein refers to an alkylene group having 1 to 4 carbon atoms.

[0044] Straight chain saturated C 1-8 An example of an alkylene group is -(CH2) n -, where n is an integer from 1 to 8, including, but not limited to, -CH2-, -CH2CH2CH2-, and -CH2CH2CH2CH2-.

[0045] Branch Saturation C 1-7 Examples of alkylene groups include, but are not limited to, -CH(CH3)-, -CH(CH3)CH2-, -CH(CH3)CH2CH2-, -CH(CH3)CH2CH2CH2-, -CH2CH(CH3)CH2-, -CH2CH(CH3)CH2-, -CH2CH(CH3)CH2CH2-, -CH(CH2CH3)-, -CH(CH2CH3)CH2-, and -CH2CH(CH2CH3)CH2-.

[0046] Linear partially unsaturated C 1-7 Examples of alkylene groups include, but are not limited to, -CH=CH-, -CH=CH-CH2-, -CH=CH-CH2-CH2-, -CH=CH-CH2CH2CH2-, -CH=CH-CH=CH-, -CH=CH-CH=CH-CH2-, -CH=CH-CH=CH-CH2-CH2-, -CH=CH-CH2-CH=CH-, and -CH=CH-CH2-CH2-CH2-CH=CH-.

[0047] Branched Partially Unsaturated C 1-7 Examples of alkylene groups include, but are not limited to, -C(CH3)=CH-, -C(CH3)=CH-CH2-, and -CH=CH-CH(CH3)-.

[0048] Alicyclic saturated C 1-7 Examples of alkylene groups include, but are not limited to, cyclopentylene (eg, cyclopent-1,3-ylene), and cyclohexylene (eg, cyclohex-1,4-ylene).

[0049] Alicyclic partially unsaturated C 1-7 Examples of alkylene groups include, but are not limited to, cyclopentenylene (eg, 4-cyclopenten-1,3-ylene), cyclohexenylene (eg, 2-cyclohexen-1,4-ylene; 3-cyclohexen-1,2-ylene; 2,5-cyclohexadien-1,4-ylene).

[0050] As used herein, the term "arylene" refers to a bidentate moiety obtained by removing two hydrogen atoms, one from each of two different aromatic ring atoms of an aromatic compound, which moiety (unless otherwise specified) has 3 to 20 ring atoms. Preferably, each ring has 5 to 7 ring atoms.

[0051] As used herein, the term "absent" used to define a variable such as X means that the variable is absent, and therefore two groups connected via the variable are directly connected to each other. For example, in -NX-Cy, when X is absent, Cy and N are directly bonded to each other.

[0052] The terms "optionally" or "optionally" mean that the subsequently described event or circumstance may, but need not, occur, and that the description includes situations where the event or circumstance occurs and situations where it does not occur. For example, "a heterocyclyl group optionally substituted with an alkyl group" means that the alkyl may, but need not, be present, and the description includes situations where the heterocyclyl group is substituted with an alkyl group and situations where the heterocyclyl group is not substituted with an alkyl group. Isomers

[0053] The present invention provides novel compounds of Formula I, or pharmaceutically acceptable salts thereof, as EP4 receptor antagonists useful in the treatment of PGE2-mediated diseases or conditions. [ka]

[0054] It is understood that certain compounds of formula I (or salts, prodrugs or conjugates) can exist and be isolated in isomeric forms, including tautomeric forms, cis isomers or trans isomers, and optically active, racemic or diastereomeric forms.The present invention encompasses the compounds of formula I in any form or mixture thereof of tautomeric forms; or as a mixture of diastereomers and as individual diastereomeric forms, and the present invention also encompasses the compounds of formula I in the form of a mixture of enantiomers and as individual enantiomers, and it is understood that any of these mixtures or forms have antagonistic properties to EP4 receptor.The methods for preparing or isolating certain forms and determining antagonistic properties to EP receptor by standard tests, including those described herein below, are well known in the art.

[0055] Furthermore, compounds of Formula I (or salts, prodrugs, or conjugates thereof) may exhibit polymorphism or form solvates with water or organic solvents. The present invention encompasses such polymorphic forms, any solvates, or any mixtures thereof.

[0056] The compounds of formula I contain one or more asymmetric centers and can therefore occur as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures, and individual diastereomers, and the present invention is intended to encompass all such isomeric forms of the compounds of formula I.

[0057] Alternatively, any enantiomer of a compound of Formula I may be obtained by stereospecific synthesis using optically pure starting materials or reagents of known configuration. salt

[0058] The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids, including inorganic or organic bases and inorganic or organic acids, which, within the scope of sound medical judgment, are suitable for use in contact with patient tissues without undue toxicity, irritation, allergic response, or other problem or complication commensurate with a reasonable benefit / risk ratio, and are effective for their intended use. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, iron(III), ferrous(II), lithium, magnesium, manganic salts, manganese, potassium, sodium, and zinc, and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium, and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, substituted amines including naturally occurring substituted amines, cyclic amines, and basic ion exchange resins, such as arginine, betaine, caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methyl-glucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, and the like.

[0059] When the compound of the present invention is basic, salts can be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, isethionic acid, lactic acid, maleic acid, malic acid, mandelic acid, methanesulfonic acid, mucic acid, nitric acid, pamoic acid, pantothenic acid, phosphoric acid, succinic acid, sulfuric acid, tartaric acid, p-toluenesulfonic acid, etc. Particularly preferred are citric acid, hydrobromic acid, hydrochloric acid, maleic acid, phosphoric acid, sulfuric acid, and tartaric acid.

[0060] It will be understood that, as used herein, references to the compounds of Formula I are meant to also include the pharmaceutically acceptable salts. Prodrug

[0061] As used herein, the term "prodrug" refers to a drug that is converted into the parent drug in vivo. Prodrugs are often useful in some situations because they may be easier to administer than the parent drug. For example, they may be orally bioavailable, while the parent drug is not. A prodrug may also have improved solubility in pharmaceutical compositions compared to the parent drug. Examples of prodrugs include, but are not limited to, compounds of Formula I, which are administered as esters ("prodrugs") to facilitate transport across cell membranes where water solubility is detrimental, but which are metabolically hydrolyzed to the active carboxylic acid once inside cells where water solubility is beneficial. Further examples of prodrugs, again without intending to limit the scope of the term, may include short peptides attached to acidic groups that are converted into the active moiety inside cells.

[0062] The present invention also encompasses acceptable forms of prodrugs of compounds of Formula I, which are formed in conventional manner with functional groups of the compounds such as amino, hydroxy, or carboxy groups. Utilities

[0063] The compounds of the present invention are EP4 receptor antagonists and are therefore expected to be useful in the treatment of diseases mediated by the EP4 receptor.

[0064] In one aspect, the invention encompasses a method for treating a human or animal subject suffering from a condition mediated by the action of PGE2 at the EP4 receptor, the method comprising administering to the subject an effective amount of a compound of formula I.

[0065] In another aspect, the invention encompasses the use of a compound of Formula I for the manufacture of a medicament for the treatment of a disease or condition mediated by the action of PGE2 at the EP4 receptor.

[0066] As used herein, the term "treatment of a prostaglandin E2 (PGE2)-mediated disease or condition" or "treatment of a disease or condition mediated by the action of PGE2 at the EP4 receptor" refers to the treatment or prevention of a chronic disease or condition that is advantageously treated or prevented by a selective EP4 antagonist. The term includes the relief of pain, fever, and inflammation from a variety of conditions, including rheumatic fever, symptoms associated with influenza or other viral infections, the common cold, back pain, neck pain, dysmenorrhea, headaches, migraines, toothaches, sprains, myositis, neuralgia, synovitis, arthritis including rheumatoid arthritis, degenerative joint diseases (osteoarthritis), gout, ankylosing spondylitis, bursitis, burns, trauma, and post-operative pain and inflammation. Furthermore, such compounds may inhibit neoplastic transformation of cells and metastatic tumor growth, and thus may be used in the treatment and / or prevention of cancer.

[0067] Examples of cancer include, but are not limited to, breast cancer, cancers that may be associated with Li-Fraumeni syndrome, such as childhood sarcoma, leukemia, and brain tumors, cancers that may be associated with Lynch syndrome, such as colon cancer, bile duct cancer, brain tumors, endometrial cancer, kidney cancer, ovarian cancer, pancreatic cancer, small intestine cancer, stomach cancer, and ureter cancer, lung cancer, melanoma, prostate cancer, retinoblastoma, thyroid cancer, and uterine cancer. Additionally, cancer may be the result of acquired mutations, such as mutations due to diet, environment, and / or lifestyle, or somatic mutations. Examples of such cancers include adrenal gland cancer, adrenocortical carcinoma, bladder cancer, brain tumor, primary brain cancer, glioma, glioblastoma, breast cancer, cervical cancer, colon cancer (non-limiting examples include colorectal cancer such as colon adenocarcinoma and colon adenoma), endometrial cancer, epidermal cancer, esophageal cancer, gallbladder cancer, genitourinary cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer (non-limiting examples include adenocarcinoma, small cell lung cancer, and non-small cell lung cancer), lymphoma (non-limiting examples include B-cell lymphoma, T-cell lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma), melanoma, malignant melanoma, malignant carcinoid cancer, malignant pancreatic insulinoma, myeloma, multiple myeloma, ovarian cancer, pancreatic cancer (such as exocrine pancreatic cancer), prostate cancer, renal cell carcinoma, skin cancer, including squamous cell carcinoma, in addition to those previously mentioned. , gastric cancer, testicular cancer, thyroid cancer, follicular thyroid carcinoma, Wilms' tumor, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, hairy cell lymphoma, Burkett's lymphoma, acute myeloid leukemia, chronic myelogenous leukemia, myelodysplastic syndrome, promyelocytic leukemia, chronic granulocytic leukemia, acute granulocytic leukemia, fibrosarcoma, rhabdomyosarcoma, astrocytoma, neuroblastoma, rhabdomyosarcoma, schwannoma, Kaposi's sarcoma, polycythemia vera, essential thrombocytosis, Hodgkin's disease, non-Hodgkin's lymphoma, soft tissue sarcoma, osteosarcoma, primary macroglobulinemia, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, and retinoblastoma.

[0068] As used herein, the terms "treat," "treating," or "treatment," etc., refer to any indication of success in preventing or ameliorating an injury, pathology, or condition, and include objective or subjective parameters such as relief; remission; reduction of symptoms or making the injury, pathology, or condition more tolerable to the patient; slowing the rate of degeneration or decline; making the end point of degeneration less debilitating; or improving the physical or mental health of the subject. Treating or ameliorating symptoms may be based on objective or subjective parameters, including the results of a physical examination, neurological examination, and / or psychiatric evaluation.

[0069] An "effective amount" or a "therapeutically effective amount" refers to an amount effective to treat a prostaglandin E2-mediated disease or condition or cancer, as determined through clinical trials and evaluations, patient observations, and the like. An "effective amount" may further designate an amount that causes a detectable change in biological or chemical activity. The detectable change may be detected and / or further quantified by those skilled in the art for the relevant mechanism or process. Furthermore, an "effective amount" may refer to an amount that maintains a desired physiological state, i.e., an amount that reduces or prevents a significant decline in symptoms and / or promotes improvement of symptoms. An "effective amount" may further refer to a therapeutically effective amount.

[0070] The compounds of Formula I may also be used in combination with one or more therapeutic agents selected from radiation and / or chemotherapeutic agents, antibodies against cytotoxic T-lymphocyte antigen 4 (anti-CTLA4), antibodies against programmed cell death ligand 1 (anti-PD-L1), antibodies against programmed cell death protein 1 (anti-PD1), indoleamine-2,3-dioxygenase (IDO) inhibitors, tryptophan-2,3-dioxygenase (TDO) inhibitors, and antimetabolites. Examples of these antibodies include, but are not limited to, MDX-010 (ipilimumab, Bristol-Myers Squibb), CP-675,206 (tremelimumab, Pfizer), MPDL3280A (Roche), MDX-1106 (nivolumab, Bristol-Myers Squibb), lambrolizumab (Merck), and pembrolizumab (KEYTRUDA®, Merck).

[0071] Examples of chemotherapeutic agents are aromatase inhibitors; antiestrogens, antiandrogens (especially in the case of prostate cancer) or gonadorelin agonists; topoisomerase I or topoisomerase II inhibitors; microtubule activators, alkylating agents, antitumor antimetabolites or platin compounds; compounds that target / reduce protein or lipid kinase activity or protein or lipid phosphatase activity, further antiangiogenic compounds or compounds that induce cell differentiation processes; bradykinin I receptor or angiotensin II antagonists; cyclooxygenase inhibitors, bisphosphonates, rapamycin derivatives, e.g. everolimus, heparanase inhibitors (preventing the degradation of heparan sulfate), e.g. PI-88, biological response modifiers, preferably These include, but are not limited to, lymphokines or interferons, e.g., interferons, ubiquitination inhibitors, or inhibitors that inhibit anti-apoptotic pathways; inhibitors of Ras oncogenic isoforms, e.g., H-Ras, K-Ras, or N-Ras, or farnesyltransferase inhibitors, e.g., L-744, 832, or DK8G557; telomerase inhibitors, e.g., telomestatin, protease inhibitors, matrix metalloproteinase inhibitors, methionine aminopeptidase inhibitors, e.g., bengamide or a derivative thereof, or proteasome inhibitors, e.g., PS-341, histone deacetylase inhibitors, e.g., vorinostat, MG0103, or MS275; and kinase inhibitors.

[0072] The pharmaceutical compositions of the present invention comprise a compound of Formula I, or a pharmaceutically acceptable salt thereof, as an active ingredient, and may also include a pharmaceutically acceptable carrier and, optionally, other therapeutic ingredients.

[0073] The EP4 antagonist, antibody, and / or antimetabolite may be administered to a subject by any suitable route, including orally (including via the oral cavity, and further including via oral gavage), intraperitoneally, parenterally, by inhalation spray, topically (i.e., to both the skin and mucosal surfaces, including the respiratory tract), transdermally, rectally, nasally (including via nasogastric gavage), sublingually, bucally, vaginally, or via an implanted reservoir. As used herein, the term "parenterally" includes subcutaneous, intramuscular, intradermal, intravenous, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. In certain embodiments, the EP4 antagonist, antibody, and / or antimetabolite is administered orally. In another specific embodiment, the EP4 antagonist, antibody, and / or antimetabolite is administered intravenously.

[0074] Pharmaceutical compositions containing the active ingredient may be in a form suitable for oral administration, such as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or as syrups or elixirs. Compositions intended for oral use may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents selected from the group consisting of sweeteners, flavorings, colorings, and preservatives to provide a pharmaceutically elegant and palatable formulation. Tablets may contain the active ingredient mixed with non-toxic pharmaceutically acceptable excipients suitable for the manufacture of tablets. These excipients may be inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrating agents such as corn starch or alginic acid; binders such as starch, gelatin, or acacia, and lubricants such as magnesium stearate, stearic acid, and talc. Tablets may be uncoated, but they may be coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period. For example, a time-delay material such as glyceryl monostearate or glyceryl distearate may be used. These may also be coated by the techniques described in U.S. Patent Nos. 4,256,108A, 4,166,452A, and 4,265,874A to form osmotic therapeutic tablets for controlled release.

[0075] The following examples illustrate selected embodiments of the present invention and are not intended to limit the scope of the invention. [Example]

[0076] Assays for measuring biological activity The compounds of Formula I may be tested using the following assays to determine their in vitro and in vivo activity as prostanoid antagonists or agonists, and their selectivity. The prostaglandin receptors that have been demonstrated to be active are DP, EP1, EP2, EP3, EP4, FP, IP, and TP. Example A. Stable expression of prostanoid receptors in the human embryonic kidney (HEK) 293 (ebna) cell line

[0077] Prostanoid receptor cDNAs corresponding to the full-length coding sequences are subcloned into appropriate sites of mammalian expression vectors and transfected into HEK 293(ebna) cells. HEK 293(ebna) cells expressing individual cDNAs are grown under selection, and after 2-3 weeks of growth, individual colonies are isolated using a cloning ring-based method and subsequently expanded into clonal cell lines. Example B. Prostanoid Receptor Binding Assay

[0078] Transfected HEK 293 (EBNA) cells are cultured and harvested. After cell lysis in the presence of protease inhibitors, membranes are prepared by differential centrifugation and used in receptor binding assays. Prostanoid receptor binding assays (for DP1, DP2 (CRTH2), EP1, EP2, EP3-III, EP4, FP, IP, and TP) are performed in 10 mM MES / KOH (pH 6.0) (EP, FP, and TP) or 10 mM HEPES / KOH (pH 7.4) (DP and IP) containing 1 mM EDTA, 2.5–30 mM divalent cations, and the appropriate radioligand that specifically binds to the target prostanoid receptor. Synthetic compounds are added to dimethyl sulfoxide, which is kept constant at 1% (v / v) for all incubations. The reaction is initiated by the addition of membrane protein. Nonspecific binding is determined in the presence of 10 μM of the corresponding nonradioactive prostanoid. Incubation is performed at room temperature or 30°C for 60-90 minutes and terminated by rapid filtration. Compound binding is calculated as the percent inhibition of radioligand binding after subtracting nonspecific binding from total binding. Residual specific binding at each ligand concentration is calculated and expressed as a function of ligand concentration, and a sigmoidal concentration-response curve is constructed. The compound's half-maximal inhibitory concentration, IC 50 The values ​​are calculated using the Hill equation curve fitting equation Y = D + [(AD) / (1 + C / IC50)] H

[0000] is determined by nonlinear regression analysis of the concentration-response curve using

[0000] . where Y = specific binding, A = left asymptote of the curve, D = right asymptote of the curve, C = compound concentration, and H = slope coefficient. The binding affinity of a compound is determined by calculating the equilibrium inhibition constant (Ki) from the equation Ki = InPt / 1 + [radioligand] / Kd, where Kd is the equilibrium dissociation constant of the radioligand:receptor interaction and InPt is the inflection point of the dose-response curve.

[0079] The EP4 receptor binding assay was performed at MSD Pharma Service in Taiwan under the following assay conditions: Source: Human recombinant Chem-1 cells Ligand: 1nM 3H] Prostaglandin E2 (PGE2) Vehicle: 1% DMSO Incubation time / temperature: 2 hours @ 25°C Incubation buffer: 10 mM MES, pH 6.0, 1 mM EDTA, 10 mM MgCl2 Nonspecific ligand: 10 μM prostaglandin E2 (PGE2) K D :0.69nM B max : 4.3 pmoles / mg protein Specific binding: 90% Quantitative method: Radioligand binding Criticality: >50% maximum suppression JPEG2025527765000006.jpg156170JPEG2025527765000007.jpg162170JPEG2025527765000008.jpg116170Example C. Microsome Stability Assay

[0080] To understand the metabolism of representative compounds, microsomal stability assays are performed. In vitro microsomal stability was measured using pooled liver microsomes from humans, dogs, rats, and mice. Test compounds (Table 2) were incubated with or without NADPH for 0–60 min, and the amount of remaining compound was quantified by LC-MS / MS analysis. JPEG2025527765000009.jpg142170 Example D. Pharmacokinetic (PK) Assay Test compounds were dissolved in 10% DMSO, 40% PEG-400, and 50% water and administered intravenously (iv) and orally (po) to rats. Three male Sprague-Dawley rats received a single intravenous dose (2 mg / kg) via the tail vein. Three male animals received a single po dose (5 mg / kg) via gavage tube. Blood samples were collected from the jugular vein into K2-EDTA tubes at 0.083, 0.25, 0.5, 1, 2, 4, 8, and 24 hours post-dose. After centrifugation, plasma samples were prepared by protein precipitation and detected by LC-MS / MS. Pharmacokinetic parameters were calculated using Phoenix software V8.3.4. JPEG2025527765000010.jpg147170 Example E. Prostanoid Receptor Agonist and Antagonist Assays

[0081] A whole-cell second messenger assay measuring stimulation of intracellular cAMP accumulation in HEK-293(ebna)-hEP4 cells was performed to determine whether receptor ligands were agonists or antagonists. Cells were harvested and resuspended in HBSS containing 25 mM HEPES, pH 7.4. Incubations included 0.5 mM IBMX (a phosphodiesterase inhibitor, available from Biomol). Samples were incubated at 37°C for 10 min, and the reaction was terminated before measuring cAMP levels. Ligands were added in dimethyl sulfoxide, which was kept constant at 1% (v / v; agonist) or 2% (v / v; antagonist) for all incubations. For agonists, the second messenger response was expressed as a function of ligand concentration, yielding the EC 50 Both the EC value and the maximal response compared to a PGE2 standard are calculated. For antagonists, the ability of a ligand to inhibit the agonist response is determined by its EC 70 The IC is determined by generating a dose-response curve in the presence of a PGE22 agonist at a concentration corresponding to the IC. 50 Values ​​are calculated as the concentration of ligand required to inhibit 50% of the PGE2-induced activity. Example F. Rat Paw Edema Assay

[0082] The method was the same as that described by Chan et al. (J. Pharmacol. Exp. Ther. 274: 1531-1537, 1995). Example G. Carrageenan-Induced Acute Inflammatory Hyperalgesia in Rats

[0083] The method was the same as that described by Boyce et al. (Neuropharmacology 33: 1609-1611, 1994). Example H. Adjuvant-Induced Arthritis in Rats

[0084] Female Lewis rats (approximately 146-170 g) were weighed, ear-marked, and assigned to groups with comparable weights (a negative control group without arthritis induction, a vehicle control group, a positive control group receiving indomethacin at a total dose of 1 mg / kg daily, and four groups receiving the test compound at a total dose of 0.10-3.0 mg / kg daily). Six groups of 10 rats each were injected into the hind paw with 0.5 mg of Mycobacterium butyricum dissolved in 0.1 mL of light mineral oil (adjuvant), and a negative control group of 10 rats was not injected with adjuvant. Body weight, contralateral paw volume (measured by mercury substitution plethysmography), and lateral radiographs (taken under ketamine and xylazine anesthesia) were measured before adjuvant injection (day -1) and 21 days after injection. Primary paw volume was measured before adjuvant injection (day -1) and on days 4 and 21 after injection. Rats were anesthetized with an intramuscular injection of 0.03–0.1 mL of a combination of ketamine (87 mg / kg) and xylazine (13 mg / kg) for radiographs and adjuvant injection. Radiographs of both hind paws were taken on days 0 and 21 using a Faxitron (45 kVp, 30 seconds) and Kodak X-OMAT TL film and developed in an automatic processor. A researcher blinded to the experimental treatments evaluated the radiographs for soft and hard tissue changes. The following radiographic changes are graded numerically according to severity: increased soft tissue volume (0–4), joint space narrowing or widening (0–5), subchondral erosion (0–3), periosteal reaction (0–4), osteolysis (0–4), subluxation (0–3), and degenerative changes in the joint (0–3). Specific criteria are used to grade the severity of each radiological change. The maximum score per foot was 26. Test compounds at total doses of 0.1, 0.3, 1, and 3 mg / kg / day, indomethacin at a total dose of 1 mg / kg / day, or vehicle (0.5% Methocel™ in sterile water) are administered orally (per os) twice daily (bid) for 21 days following adjuvant injection. Compounds are prepared weekly, refrigerated in the dark until use, and vortex-mixed immediately before administration. Example I. Mouse syngeneic model for measuring antitumor activity

[0085] The assay described in Spranger et al. (Journal for ImmunoTherapy of Cancer, 2014, 2:3) can be used to evaluate the synergistic effects of combining the compounds of the invention with effective amounts of an antibody against cytotoxic T-lymphocyte antigen 4 (anti-CTLA4); an antibody against programmed cell death ligand 1 (anti-PDL1); an antibody against programmed cell death protein 1 (anti-PD1); an indoleamine-2,3-dioxygenase (IDO) inhibitor; or a tryptophan-2,3-dioxygenase (TDO) inhibitor. Compound synthesis Example 1: 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoic acid Scheme 1 [ka]

[0086] To a solution of 3-bromo-2-chlorothiophene (5 g, 25.32 mmol, 1 equiv.) in THF (50 mL) was added dropwise LDA (2.5 M, 15.19 mL, 1.5 equiv.) over 45 min at −78 °C under argon. The reaction mixture was stirred at −78 °C for 1 h, then dry DMF (9.25 g, 126.59 mmol, 9.74 mL, 5 equiv.) was added over 15 min at −78 °C. After the addition was complete, the mixture was warmed to 25 °C and stirred for 45 min. The reaction was monitored by TLC. Upon completion of the reaction, aqueous citric acid was added, and the reaction mixture was stirred for 5 min. The mixture was diluted with ethyl acetate and washed with brine. The organic layer was dried over magnesium sulfate, filtered, and concentrated in vacuo. The residue was purified by column chromatography (SiO2, petroleum ether / ethyl acetate = 5 / 1). The compound 4-bromo-5-chloro-thiophene-2-carbaldehyde (4.7 g, 20.84 mmol, 82.3% yield) was obtained as a yellow solid. Step 2: Methyl (E)-2-azido-3-(4-bromo-5-chloro-2-thienyl)prop-2-enoate [ka]

[0087] To a solution of 4-bromo-5-chlorothiophene-2-carbaldehyde (18 g, 79.83 mmol, 1 equiv) and methyl 2-azidoacetate (27.56 g, 239.48 mmol, 3 equiv) in MeOH (180 mL) at 0° C. was added a solution of NaOMe in MeOH (4.36 M, 18.31 mL, 1 equiv), and after the addition was complete, the mixture was stirred at 0° C. for 2 h. The reaction was monitored by TLC, and after completion of the reaction, the reaction was quenched by adding saturated NH4Cl at 25 °C, then diluted with water and extracted with ethyl acetate (50 mL × 3). The organic layer was collected, dried over Na2SO4, filtered, and concentrated under reduced pressure to give crude methyl (E)-2-azido-3-(4-bromo-5-chloro-2-thienyl)prop-2-enoate (17 g, 52.70 mmol, 66.0% yield) as a yellow solid, which was used directly in the next step without further purification. Step 3: Methyl 3-bromo-2-chloro-4H-thieno[3,2-b]pyrrole-5-carboxylate [ka]

[0088] A mixture of methyl (E)-2-azido-3-(4-bromo-5-chloro-2-thienyl)prop-2-enoate (2.3 g, 7.13 mmol, 1 equiv) in xylene (30 mL) was degassed and purged with N three times, and then the mixture was stirred under a N atmosphere at 140 °C for 4 h. The reaction was monitored by TLC. After completion of the reaction, the solution was cooled to room temperature and allowed to stand overnight. The solid was collected by filtration and dried under vacuum to give methyl 3-bromo-2-chloro-4H-thieno[3,2-b]pyrrole-5-carboxylate (1.5 g, 5.09 mmol, 71.4% yield) as a white solid, which was used in the next step without further purification. Step 4: Methyl 3-bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylate [ka]

[0089] To a solution of methyl 3-bromo-2-chloro-4H-thieno[3,2-b]pyrrole-5-carboxylate (1.42 g, 4.82 mmol, 1 equiv.), 1-(bromomethyl)-4-(trifluoromethyl)benzene (1.15 g, 4.82 mmol, 743.45 μL, 1 equiv.) in DMF (15 mL) was added CsCO (4.71 g, 14.46 mmol, 3 equiv.). The mixture was stirred at 25 °C for 3 h. The reaction was monitored by TLC. Upon completion of the reaction, the reaction mixture was diluted with water and extracted with EtOAc (30 mL × 3). The combined organic layers were washed with brine (2 mL × 3), dried over NaSO, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO, petroleum ether / ethyl acetate = 10 / 1) to give methyl 3-bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylate (1.93 g, 4.26 mmol, 88.4% yield) as a white solid. MS (ES-API positive): 453.9 (M+2). + . Step 5: 3-Bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylic acid [ka]

[0090] To a solution of methyl 3-bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylate (1 g, 2.21 mmol, 1 equiv.) in THF (12.5 mL) and MeOH (12.5 mL), LiOH·HO (1 M, 6.63 mL, 3 equiv.) (1N aqueous solution) was added. The reaction mixture was stirred at 55 °C for 4 h. The reaction was monitored by LCMS. Upon completion of the reaction, most of the solvent was removed by concentration. The residue was diluted with water, acidified to pH 3-4 with 1N HCl, and extracted with EtOAc (15 mL × 3). The combined organic layers were dried over NaSO, filtered, and concentrated in vacuo to give 3-bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylic acid (950 mg, 2.17 mmol, 98.04% yield) as a white solid, which was used in the next step without further purification. MS (ES-API positive): 439.0 (M+2). + . Step 6: 3-Bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole [ka]

[0091] To a solution of 3-bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylic acid (800 mg, 1.82 mmol, 1 equiv.) in DMSO (8 mL), AgCO (502.90 mg, 1.82 mmol, 82.71 µL, 1 equiv.) and 2-3 drops of HOAc were added. The mixture was stirred at 120 °C under N for 16 h. The reaction was monitored by LCMS. After completion of the reaction, the reaction mixture was diluted with HO (15 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give 3-bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole (650 mg, 1.65 mmol, 90.31% yield) as a pale yellow solid, which was used in the next step without further purification. MS (ES-API positive): 396.0(M+2) + . Step 7: 2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid [ka]

[0092] To a mixture of 3-bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole (500 mg, 1.27 mmol, 1 equiv.) in THF (1.5 mL) was added lithium; chloro(isopropyl)magnesium; chloride (1.3 M, 1.27 mL, 1.3 equiv.) at −78° C. under N2. The reaction mixture was stirred at −78° C. for 30 min. Dry CO2 was then bubbled through the reaction mixture at −78° C. for 20 min. The reaction was monitored by LCMS. Upon completion of the reaction, water (8 mL) was added to quench the reaction, followed by extraction with EtOAc (10 mL × 3). The organic layer was dried over Na2SO4, filtered, and concentrated in vacuo. Crude 2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid (460 mg, 767.20 μmol, 60.55% yield, 60% purity) was obtained as a pale yellow oil, which was used directly in the next step without further purification. MS (ES-API positive): 360.0(M+1) + . Step 8: Methyl 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate [ka]

[0093] To a mixture of 2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid (5.35 g, 14.87 mmol, 1 equiv.) and TEA (4.51 g, 44.61 mmol, 6.21 mL, 3 equiv.) in DMF (60 mL) was added HATU (6.79 g, 17.85 mmol, 1.2 equiv.). The reaction mixture was stirred at room temperature for 30 minutes, and then methyl 4-(1-aminocyclopropyl)benzoate (2.84 g, 14.87 mmol, 1 equiv.) was added to the reaction mixture and stirred at room temperature overnight. The reaction was monitored by LCMS. After completion of the reaction, most of the solvent DMF was removed by concentration, the residue was diluted with EtOAc (50 mL), the solid was collected by filtration, washed with EtOAc (10 mL), and then dried in vacuo to give methyl 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (5.1 g, 9.57 mmol, 64.35% yield) as a white solid, which was used directly in the next step. MS (ES-API positive): 533.1(M+1) + . 1 H-NMR (500 MHz, DMSO-d6) δ 9.33 (s, 1 H), 7.73 (d, J=6.4 Hz, 2 H), 7.64 (br d, J=6.4 Hz, 2 H), 7.27 - 7.20 (m, 3 H), 7.07 (br d, J=6.4 Hz, 2 H), 6.50 (d, J=2.4 Hz, 1 H), 5.45 (s, 2 H), 3.82 (s, 3 H), 1.26 - 1.20 (m, 2 H), 0.99 - 0.93 (m, 2 H). Step 9: 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoic acid [ka]

[0094] To a mixture of methyl 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (5.1 g, 9.57 mmol, 1 equiv.) in THF (60 mL) and HO (40 mL) was added LiOH·HO (4.02 g, 95.69 mmol, 10 equiv.). The reaction mixture was stirred at 50 °C for 16 h. The reaction was monitored by LCMS. After completion of the reaction, most of the solvent was removed by concentration, then diluted with water and acidified with 1 N HCl to pH 4. It was then extracted with EtOAc (200 mL × 3), and the combined organic layers were dried over NaSO, filtered, and concentrated. The residue was recrystallized from EtOAc (40 mL) to give 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoic acid (3.8 g, 7.32 mmol, 76.52% yield, 100% purity) as a pale yellow solid. 1 H-NMR (400 MHz, DMSO-d6) δ 12.80 (br s, 1 H), 9.33 (s, 1 H), 7.74 (d, J=6.4 Hz, 2 H), 7.63 (d, J=6.4 Hz, 2 H), 7.30 - 7.18 (m, 3 H), 7.07 (d, J=6.4 Hz, 2 H), 6.50 (d, J=2.4 Hz, 1 H), 5.45 (s, 2 H) 1.25 - 1.18 (m, 2 H), 0.96 - 0.90 (m, 2 H). MS (ES-API positive): 519.2(M+1) + . Step 10: Sodium 4-(1-(2-chloro-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-3-carboxamido)cyclopropyl)benzoate [ka]

[0095] To a solution of methyl 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (26 g, 48.78 mmol, 1 equiv) in THF (200 mL), HO (100 mL), and MeOH (50 mL) was added NaOH (5.85 g, 146.35 mmol, 3 equiv) at 20 °C and stirred at 50 °C for 16 h. The reaction was monitored by TLC and LCMS. After completion of the reaction, most of the THF was removed by concentration, water (100 mL) was added, and the mixture was stirred at room temperature for 2 hours. The solid was then collected by filtration, and the filter cake was washed with water (100 mL × 2) and then dried in vacuo to give sodium 4-(1-(2-chloro-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-3-carboxamido)cyclopropyl)benzoate (25 g, 46.22 mmol, yield 94.74%) as a pale yellow solid. 1 H-NMR (400 MHz, DMSO-d6) δ 7.68 (d, J=6.8 Hz, 2 H), 7.62 (d, J=6.4 Hz, 2 H), 7.27 (d, J=6.8 Hz, 2 H),7.08 (d, J=2.4 Hz, 1 H), 6.98 (d, J=2.4 Hz, 2 H), 6.43 (d, J=2.4 Hz, 1 H), 5.41 (s, 2 H), 1.18 (t, J=7.2 Hz, 2 H), 0.93(t, J=7.2 Hz, 2 H). MS (ES-API positive): 519.0(M+1) + . Example 2 4-[1-[[2-(hydroxymethyl)-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoic acid Scheme 2 [ka] Step 1: 3-Bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylic acid [ka]

[0096] To a solution of methyl 3-bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylate (1 g, 2.21 mmol, 1 equiv.) in THF (12.5 mL) and MeOH (12.5 mL) was added 1 M LiOH (6.63 mL, 3 equiv.). The reaction mixture was stirred at 55 °C for 4 h. The reaction was monitored by LCMS. Upon completion of the reaction, most of the solvent was removed by concentration. The mixture was then diluted with water, acidified with 1 N HCl to pH 3-4, and extracted with EtOAc (15 mL x 3). The combined organic layers were dried over NaSO, filtered, and concentrated in vacuo to give crude 3-bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylic acid (950 mg, 2.17 mmol, 98.04% yield) as a white solid. MS (ES-API positive): 620.4 (M+1). + . Step 2: 3-Bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole [ka]

[0097] To a solution of 3-bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylic acid (800 mg, 1.82 mmol, 1 equiv.) in DMSO (8 mL) was added AgCO (502.90 mg, 1.82 mmol, 82.71 µL, 1 equiv.) and 50 mg of HOAc. The mixture was stirred at 120 °C under N for 16 h. The reaction was monitored by LCMS. Upon completion, the mixture was diluted with HO (50 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give 3-bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole (650 mg, 1.65 mmol, 90.31% yield) as a pale yellow solid, which was used in the next step without further purification. MS (ES-API positive): 396.0(M+1). + Step 3: 4-[[4-(trifluoromethyl)phenyl]methyl]-2-(triisopropylsilyloxymethyl)thieno[3,2-b]pyrrole-3-carboxylic acid [ka]

[0098] To a solution of [3-bromo-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrol-2-yl]methoxy-triisopropyl-silane (350 mg, 640.37 μmol, 1 equiv.) in THF (7 mL) was added lithium;chloro(isopropyl)magnesium (1.3 M, 985.19 μL, 2 equiv.) over 5 min at −78 °C under N. After the addition was complete, the reaction mixture was warmed to −10 °C and stirred at −10 °C for 30 min, then cooled back to −78 °C, and dry CO was bubbled through the reaction mixture at −78 °C for 20 min. The reaction was monitored by LCMS, and after completion, the reaction was quenched by adding water and extracted with EtOAc (10 mL × 3). The organic layer was dried over Na2SO4, filtered, and concentrated in vacuo to give 4-[[4-(trifluoromethyl)phenyl]methyl]-2-(triisopropylsiloxymethyl)thieno[3,2-b]pyrrole-3-carboxylic acid (326 mg, 637.13 μmol, 99.49% yield) as a yellow solid. MS (ES-API positive): 338.0(M+1) + . Step 4: Methyl 4-[1-[[4-[[4-(trifluoromethyl)phenyl]methyl]-2-(triisopropylsilyloxymethyl)thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate [ka]

[0099] To a solution of 4-[[4-(trifluoromethyl)phenyl]methyl]-2-(triisopropylsiloxymethyl)thieno[3,2-b]pyrrole-3-carboxylic acid (210 mg, 410.42 μmol, 1 equiv.) in DMF (4 mL) was added HATU (202.87 mg, 533.55 μmol, 1.3 equiv.) and TEA (83.06 mg, 820.84 μmol, 114.25 μL, 2 equiv.). The reaction mixture was stirred at 25° C. for 30 minutes. Methyl 4-(1-aminocyclopropyl)benzoate (78.48 mg, 410.42 μmol, 1 equiv.) was then added to the reaction mixture, which was then stirred at 25° C. for 12 hours. The reaction was monitored by LCMS, and after completion of the reaction, the reaction mixture was diluted with EtOAc (60 mL), washed with brine (8 mL × 5), dried over NaSO, and concentrated in vacuo to give methyl-4-[1-[[4-[[4-(trifluoromethyl)phenyl]methyl]-2-(triisopropylsiloxymethyl)thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (220 mg, 321.22 μmol, 78.27% yield) as a yellow oil, which was used directly in the next step without further purification. MS (ES-API positive): 707.3(M+1) + . Step 5: Methyl 4-[1-[[2-(hydroxymethyl)-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate [ka]

[0100] To a solution of methyl 4-[1-[[4-[[4-(trifluoromethyl)phenyl]methyl]-2-(triisopropylsiloxymethyl)thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (220 mg, 321.22 μmol, 1 equiv.) in THF (3 mL) was added TBAF (1 M, 385.47 μL, 1.2 equiv.). The reaction mixture was stirred at 25 °C for 1 h. TLC showed that the starting material was consumed; the desired compound was observed. The reaction mixture was concentrated in vacuo. The residue was purified by column chromatography (SiO, petroleum ether: EtOAc = 1:0 to 3:1) to give methyl 4-[1-[[2-(hydroxymethyl)-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (140 mg, 264.88 μmol, 82.46% yield) as a pale yellow solid. MS (ES-API positive): 551.1(M+1) + . Step 6: 4-[1-[[2-(hydroxymethyl)-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoic acid [ka]

[0101] To a solution of methyl 4-[1-[[2-(hydroxymethyl)-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (82 mg, 155.14 μmol, 1 equiv.) in THF (3 mL) and HO (1 mL) was added LiOH.HO (32.55 mg, 775.72 μmol, 5 equiv.). The reaction mixture was stirred at room temperature for 12 h. The reaction was monitored by LCMS. Upon completion of the reaction, the reaction mixture was acidified to pH 4 with 1N HCl and extracted with EtOAc (12 mL × 3). The organic layer was washed with brine (6 mL), dried over NaSO, filtered, and concentrated. The residue was purified by preparative HPLC (TFA) (column: Boston Green ODS 150 × 30 mm × 5 μm; mobile phase: [water (TFA)-ACN]; B%: 48%-68%, 10 min) followed by lyophilization to give 4-[1-[[2-(hydroxymethyl)-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoic acid (30.05 mg, 58.40 μmol, 37.65% yield) as a white solid. 1 H NMR (500 MHz, DMSO-d6) δ 12.98-12.59 (m, 1 H), 9.06- 8.93(m, 1 H), 7.75-7.66(m, 2 H), 7.64-7.56(m, 2 H), 7.21-7.10(m, 3 H), 7.07-6.98(m, 2 H), 6.52- 6.42 (m, 1 H), 5.57- 5.50 (m, 2 H), 4.77- 4.68 (m, 2 H), 1.28-1.15 (m, 2 H), 1.09-0.99 (m, 2 H). MS (ES-API positive): 497.2(M+1) + . Examples 3 and 4 4-(1-(2-fluoro-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-3-carboxamido)cyclopropyl)benzoic acid (Example 3) and 4-(1-(2-methoxy-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-3-carboxamido)cyclopropyl)benzoic acid (Example 4) [ka]

[0102] To a solution of 4-bromothiophene-2-carbaldehyde (7 g, 36.64 mmol, 1 equiv.) in CH3CN (40 mL) and water (40 mL) was added Select Fluor (19.47 g, 54.96 mmol, 1.5 equiv.). The mixture was stirred at 70 °C for 120 h. The reaction was monitored by TLC. Upon completion of the reaction, most of the solvent was removed by concentration, and the residue was diluted with water and extracted with EtOAc (3 × 100 mL). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (ISCO®; 80 g SepaFlash® silica flash column, elution with a 0-20% ethyl acetate / petroleum ether gradient @ 20 mL / min) to give 4-bromo-5-fluorothiophene-2-carbaldehyde (5 g, 23.92 mmol, 65.28% yield) as a white solid. Step 2: Methyl (E)-2-azido-3-(4-bromo-5-fluorothiophen-2-yl)acrylate [ka]

[0103] To a solution of 4-bromo-5-fluoro-thiophene-2-carbaldehyde (4 g, 19.14 mmol, 1 equiv.) and methyl 2-azidoacetate (6.61 g, 57.41 mmol, 3 equiv.) in MeOH (10 mL), NaOMe (4.36 M, 4.39 mL, 1 equiv.) was added. The mixture was stirred at 0 °C for 2 h. The reaction was monitored by LCMS. Upon completion of the reaction, the salts were removed by filtration, and the filtrate was concentrated under reduced pressure to give crude methyl (E)-2-azido-3-(4-bromo-5-fluoro-2-thienyl)prop-2-enoate (5 g, 16.33 mmol, 85.36% yield) as a yellow solid, which was used directly in the next step without further purification. Step 3: Methyl 3-bromo-2-fluoro-4H-thieno[3,2-b]pyrrole-5-carboxylate [ka]

[0104] A solution of methyl (E)-2-azido-3-(4-bromo-5-fluoro-2-thienyl)prop-2-enoate (1.2 g, 39.2 mmol) in xylene (10 mL) was stirred at 140° C. for 3 hours. The reaction was monitored by TLC and LCMS. After completion of the reaction, the solvent was concentrated and the residue was purified by preparative HPLC (TFA conditions) to give methyl 3-bromo-2-fluoro-4H-thieno[3,2-b]pyrrole-5-carboxylate (400 mg, 1.44 mmol, 36.69% yield) as a white solid. Step 4: Methyl 3-bromo-2-fluoro-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-5-carboxylate [ka]

[0105] To a solution of methyl 3-bromo-2-fluoro-4H-thieno[3,2-b]pyrrole-5-carboxylate (400 mg, 1.44 mmol, 1 equiv.) and 1-(bromomethyl)-4-(trifluoromethyl)benzene (343.81 mg, 1.44 mmol, 221.81 μL, 1 equiv.) in DMF (10 mL) was added CsCO (1.41 g, 4.32 mmol, 3 equiv.). The mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. Upon completion, the mixture was diluted with water and extracted with EtOAc (30 mL × 2). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to give methyl 3-bromo-2-fluoro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylate (600 mg, 1.38 mmol, 95.63% yield) as a brown solid, which was used directly in the next step without further purification. Step 5: 3-Bromo-2-fluoro-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-5-carboxylic acid [ka]

[0106] To a solution of methyl 3-bromo-2-fluoro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylate (600 mg, 1.38 mmol, 1 equiv.) in MeOH (4 mL), THF (4 mL), and HO (2 mL) was added LiOH (329.42 mg, 13.75 mmol, 10 equiv.). The mixture was stirred at 25 °C for 16 h. The reaction was monitored by TLC. Upon completion, the mixture was diluted with water and extracted with EtOAc (15 mL × 2). The combined organic layers were washed with brine, dried over Na SO , filtered, and concentrated under reduced pressure to give 3-bromo-2-fluoro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylic acid (200 mg, 473.72 μmol, 34.44% yield) as a white solid. Step 6: 3-Bromo-2-fluoro-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole [ka]

[0107] To a solution of 3-bromo-2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylic acid (50 mg, 119.55 μmol, 1 equiv.) in DMSO (1.5 mL) was added AgCO (32.97 mg, 119.55 μmol, 5.42 μL, 1 equiv.) and CHCOOH (717.93 μg, 11.96 μmol, 0.1 equiv.). The mixture was then stirred at 120 °C for 16 h. The reaction was monitored by LCMS. Upon completion, the mixture was diluted with water and extracted with EtOAc (15 mL × 2). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure to give 3-bromo-2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole (20 mg, 53.44 μmol, 44.70% yield) as a white solid, which was used directly in the next step without further purification. Step 7: 2-Fluoro-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-3-carboxylic acid [ka]

[0108] To a mixture of 3-bromo-2-fluoro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole (140 mg, 370.19 μmol, 1 equiv.) in THF (3 mL) under N at −78 °C, i-PrMgCl LiCl (1.3 M, 370.19 μL, 1.3 equiv.) was added. The reaction mixture was stirred at −10 °C for 20 min, then recooled to −78 °C, and dry CO was bubbled through at −78 °C for 20 min. The reaction was monitored by LCMS. After completion, the reaction was quenched by adding water and then extracted with EtOAc (10 mL × 3). The organic layer was dried over Na2SO4, filtered, and concentrated in vacuo to give 2-fluoro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid (123 mg, 358.29 μmol, 96.79% yield) as a pale yellow oil, which was used directly in the next step without further purification. Step 8: Methyl 4-(1-(2-fluoro-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-3-carboxamido)cyclopropyl)benzoate [ka]

[0109] To a solution of 2-fluoro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid (120 mg, 349.55 μmol, 1 equiv.) in DMF (2 mL), HATU (172.78 mg, 454.42 μmol, 1.3 equiv.) and TEA (70.74 mg, 699.11 μmol, 97.31 μL, 2 equiv.) were added over 30 min, followed by the addition of methyl 4-(1-aminocyclopropyl)benzoate (73.53 mg, 384.51 μmol, 1.1 equiv.). The mixture was stirred at 25 °C for 12.5 h. The reaction was monitored by LCMS. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. It was then diluted with water and extracted with EtOAc (15 mL × 2). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (SiO, petroleum ether / ethyl acetate=3 / 1) to give methyl 4-[1-[[2-fluoro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (40 mg, 77.44 μmol, yield 22.15%) as a white solid. MS (ES-API positive): 517.3(M+1) + . Step 9: 4-(1-(2-fluoro-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-3-carboxamido)cyclopropyl)benzoic acid [ka]

[0110] To a solution of methyl 4-[1-[[2-fluoro-4-[[4-(trifluoromethyl)phenyl]methyl thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (30 mg, 58.08 μmol, 1 equiv.) in THF (1 mL) and HO (0.3 mL) was added LiOH (13.91 mg, 580.83 μmol, 10 equiv.). The mixture was stirred at 40 °C for 12 h. The reaction was monitored by LCMS. After completion of the reaction, the solvent was concentrated and the residue was purified by preparative HPLC (TFA conditions). Column: Boston Green ODS 150 × 30 mm × 5 μm; Mobile phase: [water (TFA)-can]; B%: 53%-73%. After 10 minutes, the mixture was lyophilized to give 4-[1-[[2-fluoro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoic acid (20 mg, 39.80 μmol, 68.53% yield) as a white solid, and methyl 4-[1-[[2-hydroxy-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (5 mg, 9.72 μmol, 50.2% yield). 1 H NMR (500 MHz, DMSO) δ 12.78 (s, 1H), 9.20 (s, 1H), 7.77-7.75 (d, J = 8.5 Hz, 2H), 7.62 - 7.61 (d, J = 8.0 Hz, 2H), 7.23 (s, 1H), MS (ES-API positive): 503.1(M+1) + . 1H NMR (500MHz, CD3Cl) δ 7.93-7.91(d, J = 8.0 Hz, 2H), 7.47-7.45 (d, J = 8.0 Hz, 2H), 7.15 - 7.13 (d, J = 8.0 Hz, 2H), 7.03-7.02 (d, J = 8.0 Hz, 2H), 6.37 (s, 1H), 5.82(s, 2H), 4.08 (s, 3H), 1.46-1.44 (m, 2H), 1.29-1.24 (m, 2H). MS (ES-API positive): 515.1(M+1) + . Example 5: 3-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[1.1.1]pentane-1-carboxylic acid [ka] [ka]

[0111] To a solution of 3-cyanobicyclo[1.1.1]pentane-1-carboxylic acid (0.5 g, 3.65 mmol) in DMF (5 mL) was added K2CO3 (1.01 g, 7.29 mmol) and benzyl bromide (748.31 mg, 4.38 mmol, 519.66 μL). The mixture was stirred at room temperature for 16 h. The reaction was monitored by TLC. Upon completion of the reaction, the reaction mixture was diluted with water and extracted with EtOAc (10 mL × 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered, added benzyl bromide, and concentrated under reduced pressure. The residue was purified on a silica gel column (0–5% gradient of EtOAc in petroleum ether) to give 390 mg of the title product as a colorless oil. MS (ES-API positive): 227.1(M+1) + . Step 2: Benzyl 3-(1-aminocyclopropyl)bicyclo[1.1.1]pentane-1-carboxylate [ka]

[0112] To a solution of benzyl 3-cyanobicyclo[1.1.1]pentane-1-carboxylate (390 mg, 1.72 mmol) in EtO (10 mL) under a N atmosphere was added Ti(Oi-Pr) (536.52 mg, 1.89 mmol, 557.13 μL) at room temperature, then the reaction was cooled to −78 °C and EtMgBr (3 M, 1.26 mL) was added dropwise at −78 °C. After the addition was complete, the reaction was stirred at −78 °C for 1 h, then BF.EtO (487.13 mg, 3.43 mmol, 423.59 μL) was added dropwise at −78 °C, then the reaction was allowed to warm slowly to room temperature (25 °C) and stirred at 25 °C under a N atmosphere for 16 h. The reaction was monitored by LCMS, and after the addition was complete, the reaction mixture was quenched by dropwise addition of 1 M HCl at 25° C., diluted with HO, and then 10% aqueous NaOH (15 mL) was added dropwise at 25° C. until pH=7, and stirred at room temperature for 20 minutes, followed by extraction with EtOAc (20 mL×2). The combined organic layers were dried over NaSO and concentrated under reduced pressure to give 300 mg of the title product as a yellow oil, which was used directly in the next step. MS (ES-API positive): 258.3(M+1) + . Step 3: Benzyl 3-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[1.1.1]pentane-1-carboxylate [ka]

[0113] To a solution of benzyl 3-(1-aminocyclopropyl)bicyclo[1.1.1]pentane-1-carboxylate (160.94 mg, 625.43 μmol) and 2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid (150 mg, 416.96 μmol) in DMF (5 mL) was added HATU (475.62 mg, 1.25 mmol) and DIEA (80.83 mg, 625.43 μmol, 108.94 μL) at 25 °C. The mixture was stirred at 25 °C for 1 h. The reaction was monitored by LCMS. Upon completion of the reaction, the reaction mixture was diluted with water and extracted with EtOAc (15 mL × 3). The combined organic layers were washed with brine and then concentrated under reduced pressure to give 200 mg of the title product as a brown oil, which was used directly in the next step without further purification. MS (ES-API positive): 598.1(M+1) + . Step 4: 3-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[1.1.1]pentane-1-carboxylic acid [ka]

[0114] To a solution of benzyl 3-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[1.1.1]pentane-1-carboxylate (200 mg, 333.86 μmol) in MeOH (5 mL), THF (5 mL), and HO (2.5 mL) was added LiOH (79.95 mg, 3.34 mmol). The mixture was stirred at 55 °C for 2 h. The reaction was monitored by LCMS. Upon completion of the reaction, the reaction mixture was quenched with 1 M HCl until pH = 4-5, then extracted with EtOAc (15 mL, 3 times). The combined organic layer was concentrated under reduced pressure, and the residue was purified by preparative HPLC (HCl condition: column: Boston Green ODS 150 × 30 mm × 5 μm; mobile phase: [water (HCl)-ACN]; B%: 52% to 72%, 10 min) to give 34.46 mg of the title product as a white solid. 1 H NMR (CD3OD, 400 MHz) δ 8.79 (s, 1H), 7.58 (d, 2H, J=8.2 Hz), 7.10 (d, 1H, J=3.0 Hz), 7.05 (br d, 2H, J=8.1 Hz), 6.45 (d, 1H, J=3.0 Hz), 5.53 (s, 2H), 1.82 (s, 6H), 0.70-0.60 (m, 2H), 0.40 -0.30 (m, 2H). MS (ES-API positive): 509.0(M+1) + . Example 6. 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[2.2.2]octane-1-carboxylic acid [ka]

[0115] A mixture of 4-methoxycarbonylbicyclo[2.2.2]octane-1-carboxylic acid (10 g, 47.12 mmol), BOP (20.84 g, 47.12 mmol), TEA (19.07 g, 188.46 mmol, 26.23 mL), and NH₄Cl (25.20 g, 471.16 mmol) in DMF (100 mL) was stirred at 25 °C for 16 h. The reaction was monitored by LCMS. Upon completion of the reaction, the reaction mixture was diluted with water (ca. 50 mL) and extracted with EtOAc (50 mL × 3). The combined organic layers were washed with brine, dried over Na₂SO₄, filtered, and concentrated under reduced pressure to give 9.9 g of the crude title compound as a pale yellow oil, which was used directly in the next step. MS (ES-API positive): 212.3(M+1) + . Step 2: Methyl 4-cyanobicyclo[2.2.2]octane-1-carboxylate [ka]

[0116] To a solution of methyl 4-carbamoylbicyclo[2.2.2]octane-1-carboxylate (9.9 g, 46.86 mmol) in pyridine (80 mL) was added TFAA (68.90 g, 328.04 mmol, 45.63 mL) dropwise at 0 °C. The mixture was then slowly warmed to room temperature and stirred overnight. The reaction was monitored by TLC. Upon completion of the reaction, the reaction mixture was diluted with 1 N HCl and extracted with EtOAc (30 mL × 3). The combined organic layers were washed with brine, dried over Na2SO4, filtered, and concentrated under reduced pressure. The residue was purified on a silica gel column (0–20% gradient of EtOAc in petroleum ether) to give 9 g of the title product as a yellow solid. MS (ES-API positive): 193.1(M+1) + . Step 3: Methyl 4-(1-aminocyclopropyl)bicyclo[2.2.2]octane-1-carboxylate [ka]

[0117] To a solution of methyl 4-cyanobicyclo[2.2.2]octane-1-carboxylate (470 mg, 2.43 mmol) in EtO (30 mL) under a N atmosphere was added Ti(Oi-Pr) (760.38 mg, 2.68 mmol, 789.59 μL) at 25 °C, then the reaction was cooled to −78 °C and EtMgBr (3 M, 1.78 mL) was added dropwise at −78 °C. After the addition was complete, the reaction was stirred at −78 °C for 1 h, then BF.EtO (690.40 mg, 4.86 mmol, 600.35 μL) was added dropwise at −78 °C, then the reaction was slowly warmed to room temperature and stirred at 25 °C for 5 h under a N atmosphere. The reaction was monitored by LCMS, and after completion of the reaction, the reaction was quenched by adding 1M HCl dropwise at room temperature, then diluted with water, and then 10% aqueous NaOH solution (15 mL) was added dropwise at 25° C. until pH=7, and the mixture was stirred at room temperature for 20 minutes, followed by extraction with EtOAc (20 mL×2). The combined organic layer was dried over NaSO and concentrated under reduced pressure to give 350 mg of the title product as a yellow oil, which was used in the next step without purification. MS (ES-API positive): 224.3(M+1) + . Step 4: Methyl 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[2.2.2]octane-1-carboxylate [ka]

[0118] To a solution of 2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid (100 mg, 277.97 μmol) in DMF (5 mL) was added methyl 4-(1-aminocyclopropyl)bicyclo[2.2.2]octane-1-carboxylate (124.15 mg, 277.97 μmol, 50% purity), DIEA (107.77 mg, 833.91 μmol, 145.25 μL), and HATU (158.54 mg, 416.96 μmol). The mixture was stirred at 25 °C for 16 h. The reaction was monitored by LCMS. Upon completion of the reaction, the reaction mixture was diluted with HO and extracted with EtOAc (15 mL × 3). The combined organic layers were washed with brine and then concentrated under reduced pressure to give 150 mg of the title product as a brown oil, which was used directly in the next step without purification. MS (ES-API positive): 565.1(M+1) + . Step 5: 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[2.2.2]octane-1-carboxylic acid [ka]

[0119] To a solution of methyl 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[2.2.2]octane-1-carboxylate (150 mg, 265.47 μmol) in THF (5 mL), MeOH (5 mL), and HO (2.5 mL) was added LiOH (63.58 mg, 2.65 mmol). The mixture was stirred at 55 °C for 3 h and the reaction was monitored by LCMS. After completion of the reaction, the reaction mixture was diluted with HO and 1 M HCl until pH = 4–5, and then extracted with EtOAc (15 mL × 3). The combined organic layer was concentrated under reduced pressure, and the residue was purified by preparative HPLC (HCl condition: column: Boston Green ODS 150 × 30 mm × 5 μm; mobile phase: [water (HCl)-ACN]; B%: 70% to 90%, 10 min) to give 45.48 mg of the title product as a white solid. 1 H NMR (CD3OD, 400 MHz) δ 8.62 (s, 1H), 7.56 (d, 2H, J=8.0 Hz), 7.13-7.02 (m, 3H), 6.42 (d, 1H, J=3.2 Hz), 5.50 (s, 2H), 1.65-1.61(m, 6H), 1.42-1.38 (m, 6H), 0.73-0.70 (m, 2H), 0.28-0.25 (m, 2H). MS (ES-API positive): 551.1(M+1) + . Example 7: 3-[1-[[2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[1.1.1]pentane-1-carboxylic acid [ka]

[0120] To a solution of benzyl 3-(1-aminocyclopropyl)bicyclo[1.1.1]pentane-1-carboxylate (170.63 mg, 663.07 μmol) and 2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid (150 mg, 442.05 μmol) in DMF (5 mL) was added HATU (252.12 mg, 663.07 μmol) and DIEA (171.39 mg, 1.33 mmol, 230.98 μL) at 25 °C. The mixture was stirred at 25 °C for 1 h. The reaction was monitored by LCMS. Upon completion of the reaction, the reaction mixture was quenched with water and extracted with EtOAc (15 mL × 3). The combined organic layers were washed with brine and then concentrated under reduced pressure to give 200 mg of the title product as a brown oil, which was used directly in the next step without further purification. MS (ES-API positive): 579.3(M+1) + . Step 2: 3-[1-[[2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[1.1.1]pentane-1-carboxylic acid [ka]

[0121] To a solution of benzyl 3-[1-[[2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[1.1.1]pentane-1-carboxylate (0.2 g, 345.64 μmol) in MeOH (5 mL), THF (5 mL), and HO (2.5 mL) was added LiOH (82.78 mg, 3.46 mmol). The mixture was stirred at 50 °C for 2 h. The reaction was monitored by LCMS. Upon completion of the reaction, the reaction mixture was diluted with water, and then 1 M HCl was added until the pH reached 4–5. The mixture was then extracted with EtOAc (15 mL × 3). The combined organic layer was concentrated under reduced pressure, and the residue was purified by preparative HPLC (HCl condition: column: Boston Green ODS 150 × 30 mm × 5 μm; mobile phase: [water (HCl)-ACN]; B%: 50% to 70%, 10 min) to give 55.14 mg of the title product as a white solid. 1 H NMR (CD3OD, 400 MHz) δ 8.58 (s, 1H), 7.53 (d, 2H, J=8.0 Hz), 7,13-6.92 (m, 3H), 6.38 (d, 1H, J=3.2 Hz), 5.51 (s, 2H), 2.57 (s, 3H), 1.77 (s, 6H), 0.71-0.63(m, 2H), 0.42-0.33 (m, 2H). MS (ES-API positive): 489.0(M+1) + . Example 8: 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[2.2.2]octane-1-carboxylic acid [ka] Step 1: Methyl 4-[1-[[2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[2.2.2]octane-1-carboxylate [ka]

[0122] To a solution of 2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid (150 mg, 442.05 μmol) in DMF (5 mL) was added methyl 4-(1-aminocyclopropyl)bicyclo[2.2.2]octane-1-carboxylate (296.14 mg, 663.07 μmol, 50% purity), DIEA (171.39 mg, 1.33 mmol, 230.98 μL), and HATU (252.12 mg, 663.07 μmol). The mixture was stirred at 25 °C for 16 h. The reaction was monitored by LCMS. Upon completion, the reaction was quenched with water and extracted with EtOAc (15 mL × 3). The combined organic layers were washed with brine and then concentrated under reduced pressure to give 0.2 g of the title compound as a brown oil, which was used directly in the next step without further purification. MS (ES-API positive): 545.4(M+1) + . Step 2: 4-[1-[[2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[2.2.2]octane-1-carboxylic acid [ka]

[0123] To a solution of methyl 4-[1-[[2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]bicyclo[2.2.2]octane-1-carboxylate (0.2 g, 367.22 μmol) in THF (5 mL), MeOH (5 mL), and HO (2.5 mL) was added LiOH (380 mg, 15.87 mmol). The mixture was stirred at 50 °C for 3 h. The reaction was monitored by LCMS. Upon completion of the reaction, the reaction mixture was diluted with HO (10 mL), and then 1 M HCl was added until the pH reached 4–5, followed by extraction with EtOAc (15 mL × 3). The combined organic layers were concentrated under reduced pressure. The residue was purified by preparative HPLC (HCl conditions: column: Boston Green ODS 150 × 30 mm × 5 μm; mobile phase: [water (HCl)-ACN]; B%: 56% to 76%, 10 min) to give 14.59 mg of the title product as a white solid. 1 H NMR (CD3OD, 400 MHz) δ 8.47 (s, 1H), 7.57 (d, 2H, J=8.0 Hz), 7.12-6.93 (m, 3H), 6.39 (d, 1H, J=3.2 Hz), 5.53 (s, 2H), 2.57 (s, 3H), 1.72-1.64 (m, 6H), 1.43 -1.36 (m, 6H), 0.82 -0.75 (m, 2H), 0.33-0.28 (m, 2H). MS (ES-API positive): 531.0(M+1) + . Example 9: 3-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]ethyl]bicyclo[1.1.1]pentane-1-carboxylic acid [ka]

[0124] To a solution of 3-methoxycarbonylbicyclo[1.1.1]pentane-1-carboxylic acid (1.0 g, 5.88 mmol, 1 equiv.), N-methoxymethanamine hydrochloride (687.89 mg, 7.05 mmol, 1.2 equiv.), and EtN (1.78 g, 17.63 mmol, 2.45 mL, 3.0 equiv.) in DCM (15 mL) was added HATU (2.68 g, 7.05 mmol, 1.2 equiv.). The mixture was stirred at room temperature overnight. The reaction was monitored by LCMS. Upon completion, the reaction was quenched with water and then extracted with DCM (10 mL x 3). The combined organic layers were concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (ISCO®, 12 g SepaFlash® silica flash column, elution with a 0-25% ethyl acetate / petroleum ether gradient @ 30 mL / min) to give 950 mg of the title product as a white solid. MS (ES-API positive): 214.2 (M+1). + . Step 2: Methyl 3-acetylbicyclo[1.1.1]pentane-1-carboxylate [ka]

[0125] To a solution of methyl 3-[methoxy(methyl)carbamoyl]bicyclo[1.1.1]pentane-1-carboxylate (200 mg, 937.96 μmol, 1 equiv.) in THF (10 mL) was added MeMgBr (3 M, 468.98 μL, 1.5 equiv.) at −78 °C. After the addition was complete, the reaction mixture was slowly warmed to room temperature and stirred at room temperature for 2 h. The reaction was monitored by TLC. Upon completion, the reaction mixture was quenched by the addition of saturated NH4Cl, then diluted with water and extracted with ethyl acetate (10 mL × 3). The combined organic layers were concentrated under reduced pressure. The residue was purified by flash silica gel chromatography (ISCO®, 4 g SepaFlash® silica flash column, elution with a 0–20% ethyl acetate / petroleum ether gradient at 30 mL / min) to afford 70 mg of the title product as a white solid. Step 3: Methyl 3-[(Z)-N-hydroxy-C-methyl-carbonimidoyl]bicyclo[1.1.1]pentane-1-carboxylate [ka]

[0126] A mixture of methyl 3-acetylbicyclo[1.1.1]pentane-1-carboxylate (70 mg, 416.20 μmol, 1 equiv.), AcONa (102.43 mg, 1.25 mmol, 3 equiv.), and hydroxylammonium chloride (86.77 mg, 1.25 mmol, 3 equiv.) in MeOH (5 mL) was stirred at 65 °C for 2 h under a N atmosphere. The reaction was monitored by LCMS. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove MeOH. The residue was diluted with ethyl acetate and washed with brine (10 mL × 2). The organic layer was concentrated under reduced pressure to give 70 mg of the title product as a pale white solid, which was used directly in the next step without further purification. MS (ES-API positive): 184.1(M+1) + Step 4: Methyl 3-(1-aminoethyl)bicyclo[1.1.1]pentane-1-carboxylate [ka]

[0127] A mixture of methyl 3-[(Z)-N-hydroxy-C-methyl-carbonimidoyl]bicyclo[1.1.1]pentane-1-carboxylate (70 mg, 382.09 μmol, 1 equiv.) and Raney-Ni (51.07 mg, 596.06 μmol, 1.56 equiv.) in MeOH (3 mL) was degassed and purged with H three times, and then the mixture was stirred under an H atmosphere at 25 °C for 15 h. The reaction was monitored by LCMS. Upon completion of the reaction, the catalyst was removed by filtration, and the filtrate was concentrated under reduced pressure to give 60 mg of the title product as a pale colorless oil, which was used directly in the next step without further purification. MS (ES-API positive): 170.2(M+1) + . Step 5: Methyl 3-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]ethyl]bicyclo[1.1.1]pentane-1-carboxylate [ka]

[0128] To a solution of methyl 3-(1-aminoethyl)bicyclo[1.1.1]pentane-1-carboxylate (60 mg, 354.57 μmol, 1 equiv.) in DMF (5 mL), 2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid (127.56 mg, 354.57 μmol, 1 equiv.), HATU (161.78 mg, 425.48 μmol, 1.2 equiv.), and DIPEA (137.48 mg, 1.06 mmol, 185.28 μL, 3 equiv.) were added. The mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. Upon completion, the reaction mixture was quenched with water and extracted with ethyl acetate (3 × 10 mL). The combined organic layers were concentrated under reduced pressure to give 150 mg of the title product as a brown oil, which was used directly in the next step without further purification. MS (ES-API positive): 511.2(M+1) + . Step 6: 3-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]ethyl]bicyclo[1.1.1]pentane-1-carboxylic acid [ka]

[0129] To a solution of methyl 3-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]ethyl]bicyclo[1.1.1]pentane-1-carboxylate (150 mg, 293.57 μmol, 1 equiv.) in THF (1 mL), MeOH (1 mL), and HO (0.5 mL) was added LiOH (70.30 mg, 2.94 mmol, 10 equiv.). The mixture was stirred at 50 °C for 2 h. The reaction was monitored by LCMS. After completion of the reaction, the reaction mixture was concentrated under reduced pressure to remove the solvent, then diluted with water, adjusted to pH 4-5 with 1 N HCl (aq.), and extracted with ethyl acetate (3 × 10 mL). The combined organic layers were washed with brine and concentrated under reduced pressure. The residue was purified by reverse-phase HPLC (column: Boston Green ODS 150×30 mm×5 μm; mobile phase: [water (HCl)-ACN]; B%: 52% to 72%, 10 min) to give 61.2 mg of the title product as a pale white solid. 1 H NMR (400 MHz, CD3OD) δ 8.32 (br d, J=8.0 Hz, 1H), 7.56 (d, J=8.4 Hz, 2H), 7.14-7.10 (m, 3H), 6.43 (d, J=2.8 Hz, 1H), 5.58-5.40 (m, 2H), 4.02-3.95 (m, 1H), 1.94-1.84 (m, 6H), 0.83 (d, J=6.8 Hz, 3H). MS (ES-API positive): 497.0(M+1) + . Example 10: 3-[1-[[2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]ethyl]bicyclo[1.1.1]pentane-1-carboxylic acid Scheme 9 [ka]

[0130] To a solution of methyl 3-(1-aminoethyl)bicyclo[1.1.1]pentane-1-carboxylate (50 mg, 295.47 μmol, 1 equiv.) and 2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid (100.26 mg, 295.47 μmol, 1 equiv.) in DMF (3 mL), HATU (134.82 mg, 354.56 μmol, 1.2 equiv.) and DIPEA (114.56 mg, 886.41 μmol, 154.40 μL, 3 equiv.) were added. The mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. Upon completion, the reaction was quenched with water and extracted with ethyl acetate (3 × 10 mL). The combined organic layers were concentrated under reduced pressure to give 140 mg of the title product as a white solid, which was used directly in the next step without further purification. MS (ES-API positive): 491.2(M+1) + . Step 2: 3-[1-[[2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]ethyl]bicyclo[1.1.1]pentane-1-carboxylic acid [ka]

[0131] To a solution of methyl 3-[1-[[2-methyl-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]ethyl]bicyclo[1.1.1]pentane-1-carboxylate (140 mg, 285.40 μmol, 1 equiv.) in THF (2 mL), MeOH (2 mL), and water (1 mL), LiOH (68.35 mg, 2.85 mmol, 10 equiv.) was added. The mixture was stirred at 50 °C for 2 h. The reaction was monitored by LCMS. Upon completion of the reaction, most of the solvent was removed by concentration. The residue was diluted with water, adjusted to pH 4-5 with 1 N HCl (aq.), and then extracted with ethyl acetate (10 mL × 3). The combined organic layers were washed with brine and then concentrated under reduced pressure, and the residue was purified by reverse-phase HPLC (column: Boston Green ODS 150 × 30 mm × 5 μm; mobile phase: [water (HCl)-ACN]; B%: 50% to 70%, 10 min) to give 47.7 mg of the title product as a pale white solid. 1 H NMR (400 MHz, CD3OD) δ 8.11 (br d, J=8.0 Hz, 1H), 7.54 (d, J=8.0 Hz, 2H), 7.08 (d, J=8.0 Hz, 2H), 6.99 (d, J=3.2 Hz, 1H), 6.38 (d, J=3.2 Hz, 1H), 5.60-5.40 (m, 2H), 4.03-3.96 (m, 1H), 2.56 (s, 3H), 1.82 (s, 6H), 0.84 (d, J=6.8 Hz, 3H). MS (ES-API positive): 477.1(M+1) + . Example 11: 4-(1-(2-(trifluoromethyl)-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-3-carboxamido)cyclopropyl)benzoic acid [ka]

[0132] 2-Chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid (0.2 g, 555.94 μmol, 1 equiv.), LiCl (47.14 mg, 1.11 mmol, 22.77 μL, 2 equiv.), TMSCl (2.42 mg, 22.24 μmol, 2.82 μL, 0.04 equiv.), dibromocobalt (12.16 mg, 55.59 μmol, 0.1 equiv.), 4,7-diphenyl-1-methyl-2-pyrrole (1.0 g, 555.94 μmol, 1 equiv.) in THF (5 mL). A mixture of 10-phenanthroline (18.48 mg, 55.59 μmol, 0.1 equiv.), 167.98 μL, 1.5 equiv.) and indigane (255.33 mg, 2.22 mmol, 34.98 μL, 4 equiv.) was stirred at 80°C for 20 hours. The reaction was then cooled to room temperature, and molecular iodine (211.65 mg, 833.91 μmol) was added and stirred at room temperature overnight. The reaction was monitored by LCMS. After completion of the reaction, the salt was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC (FA conditions: column: Boston Green ODS 150 × 30 mm × 5 μm; mobile phase: [water (FA)-ACN]; B%: 62% to 92%, 12 min) to obtain 2-iodo-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid (42 mg, 93.08 μmol, yield 16.74%) as a pale pink solid. MS (ES-API positive): 451.9(M+1) + . Step 2: Methyl 4-(1-(2-iodo-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-3-carboxamido)cyclopropyl)benzoate [ka]

[0133] To a solution of 2-iodo-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carboxylic acid (42 mg, 93.08 μmol, 1 equiv.) and methyl 4-(1-aminocyclopropyl)benzoate (26.70 mg, 139.63 μmol, 1.5 equiv.) in DMF (2 mL) was added HATU (106.18 mg, 279.25 μmol, 3 equiv.) and DIEA (18.05 mg, 139.63 μmol, 24.32 μL, 1.5 equiv.). The mixture was stirred at 25 °C for 2 h. The reaction was monitored by LCMS. Upon completion, the reaction was quenched by adding water, followed by extraction with ethyl acetate (10 mL × 3). The organic layer was collected and concentrated under reduced pressure. The residue was purified by preparative HPLC (FA conditions; column: Boston Green ODS 150 × 30 mm × 5 μm; mobile phase: [water (FA)-ACN]; B%: 53% to 83%, 12 min) to obtain methyl 4-[1-[[2-iodo-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (26 mg, 41.64 μmol, yield 44.73%) as a white solid. MS (ES-API positive): 625.0(M+1) + . Step 3: Methyl 4-(1-(2-(trifluoromethyl)-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-3-carboxamido)cyclopropyl)benzoate [ka]

[0134] To a solution of methyl 4-[1-[[2-iodo-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (16 mg, 25.62 μmol, 1 equiv.) in DMF (1.5 mL), CuI (14.64 mg, 76.87 μmol, 3 equiv.) and methyl 2,2-difluoro-2-fluorosulfonyl acetate (14.77 mg, 76.87 μmol, 9.78 μL, 3 equiv.) were added. The mixture was then stirred at 100 °C for 16 h. The reaction was monitored by LCMS. Upon completion of the reaction, the salts were removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by preparative HPLC (FA conditions: column: Boston Green ODS 150 × 30 mm × 5 μm; mobile phase: [water (FA)-ACN]; B%: 65% to 95%, 12 min) to obtain methyl 4-[1-[[2-(trifluoromethyl)-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (8 mg, 14.12 μmol, yield 55.11%) as a white solid. MS (ES-API positive): 567.2(M+1) + . Step 4: 4-(1-(2-(trifluoromethyl)-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-3-carboxamido)cyclopropyl)benzoic acid [ka]

[0135] To a solution of methyl 4-[1-[[2-(trifluoromethyl)-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (11 mg, 19.42 μmol, 1 equiv.) in MeOH (1 mL), THF (1 mL), and HO (0.5 mL) was added LiOH (4.65 mg, 194.17 μmol, 10 equiv.). The mixture was stirred at 55 °C for 1 h. The reaction was monitored by LCMS. Upon completion, the reaction was quenched by adding 1 M HCl until pH = 3-4, then diluted with water and extracted with EtOAc (3 × 10 mL). The organic layer was collected and concentrated under reduced pressure. The residue was purified by preparative HPLC (HCl condition; column: Boston Green ODS 150 × 30 mm × 5 μm; mobile phase: [water (HCl)-ACN]; B%: 56% to 76%, 10 min) to obtain 4-[1-[[2-(trifluoromethyl)-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoic acid (3 mg, 5.43 μmol, yield 27.97%) as a white solid. 1 H NMR (400 MHz, CD3OD) δ 9.48 (s, 1H), 7.67 (d, J=8.4 Hz, 2H), 7.43 (d, J=8.4 Hz, 2H), 7.26 (d, J=8.4 Hz, 2H), 7.03 (d, J=2.8 Hz, 1H), 6.88 (d, J=8.4 Hz, 2H), 6.46 (d, J=2.8 Hz, 1H), 5.16 (s, 2H), 1.15-1.12 (m, 2H), 1.01-0.98 (m, 2H). MS (ES-API positive): 553.2(M+1) + .

[0136] The above examples and descriptions are not intended to limit the scope of the present invention. Any combination of the embodiments of the present invention, and any obvious extensions or similar forms, are within the scope of the present invention. Moreover, the present invention is intended to encompass any arrangement calculated to achieve the same purpose, and all such variations and modifications are intended to be included within the scope of the appended claims.

Claims

1. Compound of formula I, 【Chemistry 1】 [In the formula: R １ , １－６ and R ２ are each independently hydrogen, C １－６ alkyl, C １－６ cycloalkyl, C １－６ halocycloalkyl, or C １－６ haloalkyl; or, R １ and R ２ together with the carbon atom to which they are both attached form a 3- to 6-membered carbocyclic ring, said carbocyclic ring being optionally substituted with 1 to 3 R ａ groups and optionally containing 1 or 2 ring-forming heteroatom(s) each independently being S, O, or NR ｂ and each Rb is independently hydrogen, C １－６ alkyl, C １－６ cycloalkyl, C １－６ halocycloalkyl, and C １－６ haloalkyl, aryl, heteroaryl, -C(O)-C １－６ alkyl, -C(O)-aryl, -S(O) ２ -alkyl, or -S(O) ２ -aryl; X does not exist, =CH-, -CR １ R ２ - or -C(O)-; Cy １ However, C １－６ Alkylene, C １－６ Alkenylene, C １－６ Alkynylene, cycloalkylene, cycloalkenylene, arylene, heteroarylene, heterocyclylene, or cross-linked bicyclic cycloalkylene or cycloalkenylene, C １－６ Alkylene, C １－６ Alkenylene, C １－６ Each of the following can be optionally substituted: alkynylene, cycloalkylene, cycloalkenylene, arylene, heteroarylene, heterocyclylene, or bridged bicyclic cycloalkylene; Cy ２ However, the substituents are cycloalkyl, aryl, heteroaryl, or heterocyclyl, each independently substituted with one to three substituents that are halo, alkyl, or haloalkyl; Each R ａ The group is independently halo, alkyl, haloalkyl, hydroxyalkyl, or alkoxy; R ａ If it is alkyl, Cy １ This is a cross-linked bicyclic cycloalkylene. or a pharmaceutically acceptable salt thereof.

2. X is -CH ２ - The compound according to claim 1.

3. Cy ２ The compound according to claim 1, wherein the compound is aryl and optionally substituted with one haloalkyl group.

4. The compound according to claim 1, wherein the halo is -F or -Cl.

5. Cy １ The compound according to claim 1, wherein the compound is an arylene or a crosslinked bicyclic cycloalkylene.

6. R １ and R ２ Each of them independently, hydrogen or C １ - ６ Alkyl; or R １ and R ２ The compound according to claim 1, wherein both of them, together with the carbon atom to which they are bonded, form a 3- to 6-membered carbocyclic ring.

7. R ａ -F, -Cl, -CF ３ , hydroxyalkyl, alkoxyl or -CH ３ And; R ａ ga-CH ３ In the case of Cy １ C ５ -C １０ The compound according to claim 1, which is a crosslinked bicyclic cycloalkylene.

8. The compound according to claim 1, wherein the compound is of formula II. 【Chemistry 2】 II [In the formula, Cy １ is an arylene or a crosslinked bicyclic cycloalkylene; R ａ R is a halo, alkyl, haloalkyl, hydroxyalkyl, or alkoxy; ａ If it is alkyl, Cy １ This is a cross-linked bicyclic cycloalkylene.

9. Cy １ However, phenylene or C ５ -C １０ The compound according to claim 8, which is a crosslinked bicyclic cycloalkylene.

10. Said C ５ -C １０ Cross-linked bicyclic cycloalkylenes, 【Transformation 3】 【Chemistry 4】 The compound according to claim 9.

11. The aforementioned compound, 【Transformation 5】 The compound according to claim 1.

12. A pharmaceutical composition comprising the compound described in claim 1 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or excipient.

13. The pharmaceutical composition according to claim 12, further comprising another therapeutic agent selected from the group consisting of an antibody against cytotoxic T lymphocyte antigen 4 (anti-CTLA4), an antibody against programmed cell death ligand 1 (anti-PDL1), an antibody against programmed cell death protein 1 (anti-PD1), an indoleamine-2,3-dioxygenase (IDO) inhibitor, and tryptophan-2,3-dioxygenase (TDO) inhibitors and antimetabolites.

14. The pharmaceutical composition according to claim 12, wherein the composition is used in combination with a radiotherapy agent.

15. A method for treating a subject suffering from symptoms mediated by the action of PGE2 at the EP4 receptor, comprising administering an effective amount of the compound described in claim 1 to the subject in need thereof.

16. The method according to claim 15, wherein the symptoms are an inflammatory disease or cancer.

17. The method according to claim 16, wherein the inflammatory disease is arthritis, acne vulgaris, asthma, autoimmune disease, autoinflammatory disease, celiac disease, chronic prostatitis, colitis, diverticulitis, glomerulonephritis, hidradenitis suppurativa, hypersensitivity, inflammatory bowel disease, interstitial cystitis, mast cell activation syndrome, macrocytosis, otitis, pelvic inflammatory disease, reperfusion injury, rheumatic fever, rheumatoid arthritis, rhinitis, sarcoidosis, or vasculitis.

18. The method according to claim 16, wherein the cancer is breast cancer, endometrial cancer, cervical cancer, ovarian cancer, lung cancer, head and neck cancer, brain cancer, thyroid cancer, esophageal cancer, stomach cancer, colorectal cancer, liver cancer, pancreatic cancer, skin cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, bone cancer, lymphoma, or hematological cancer.

19. Use of the compound according to claim 1 for the manufacture of a pharmaceutical product for treating a subject suffering from symptoms mediated by the action of PGE2 at the EP4 receptor.

20. The use according to claim 19, wherein the symptoms are pain, inflammatory disease, and cancer.

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