Heterobicyclic compounds as EP4 receptor antagonists

Abstract

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

Description

Cross - Reference to Related Applications

[0001] This application claims priority to International Application No. PCT / CN2022 / 114402, filed on August 24, 2022, and U.S. Application No. 63 / 425,555, filed on November 15, 2022, the entire contents of both of which are incorporated herein by reference. Background Art

[0002] The present invention relates to heterocyclic amide derivatives, or pharmaceutically acceptable salts, pharmaceutically acceptable prodrugs thereof, pharmaceutical compositions made therefrom, and the medical use of the above substances in mammals (including humans). The compounds of the present invention have activity as prostaglandin E2 (PGE2) receptor antagonists and can be used to treat or relieve pain, inflammation, and cancer.

[0003] Prostaglandins are a group of physiologically active lipid compounds that have various hormone - like effects in the body. Prostaglandins are interesting because they produce different effects depending on the receptors to which they attach. Studies have shown that prostaglandins are important mediators of pain, fever, and other inflammation - related symptoms. Of particular interest is prostaglandin E2 (PGE2), which is the major eicosanoid associated with inflammation. In addition, highly expressed PGE2 in tumor tissues inhibits anti - tumor immunity in the tumor microenvironment (TME) and leads to tumor immune escape, thus resulting in disease progression (see Front Immunol. 2020; 11:324). PGE2 has also been shown to promote the expansion and metastasis of mouse colorectal cancer stem cells (CSCs) (Wang et al., Gastroenterology, 2015, 1 - 12). The activity of PGE2 is mainly mediated by binding to a group of G - protein - coupled receptors (GPCRs), namely the E - type prostaglandin (EP) receptors EP1, EP2, EP3, and EP4. Among them, the EP4 receptor is the most versatile and promising emerging receptor among current PGE2 receptors. In animal studies, selective inhibition of PGE2 / EP4 receptor signaling by antagonists reduced tumor growth (Terada et al. Cancer Res. 2010, 70, 1606 - 1615) and tumor metastasis (Yang et al. Cancer Res. 2006, 66, 9665 - 9672).

[0004] To date, EP4 receptor antagonists of different structural classes have been described. The object of the present invention is to provide a series of novel heterobicyclic compounds as EP4 receptor antagonists, and methods for treating diseases or disorders mediated by the action of PGE2 on the EP4 receptor (including pain, inflammation, and cancer), and their pharmaceutical compositions. Summary of the Invention

[0005] The present invention relates to a series of novel heterobicyclic amide derivatives as EP4 receptor antagonists, which can be used to treat diseases or disorders mediated by the action of PGE2 on the EP4 receptor, such as pain, inflammation and cancer. Also included are pharmaceutical compositions and methods of use.

[0006] In one aspect, the present invention provides a heterobicyclic compound or a pharmaceutically acceptable salt thereof. The compound has the following structural formula I: Wherein: R 1 and R 2 are each 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 atom to which they are attached form a 3- to 6-membered carbocyclic ring, which carbocyclic ring is optionally substituted with one to three R a groups and optionally contains one or two ring-forming heteroatoms, each ring-forming heteroatom independently being S, O or NR b , wherein each R b 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 is absent, =CH-, -CR 1 R 2 - or -C(O)-; Cy 1 is C 1-6 alkylene, C 1-6 alkenylene, C 1-6 alkynylene, cycloalkylene, arylene, heteroarylene, heterocyclylene, or bridged bicyclic cycloalkylene, and is optionally substituted; Cy 2 is cycloalkyl, aryl, heteroaryl or heterocyclic group, and each of the cycloalkyl, aryl, heteroaryl or heterocyclic group is optionally substituted with one to three substituents, each substituent independently being halogen, alkyl or haloalkyl; and each R a is independently halogen, alkyl, haloalkyl, hydroxyalkyl or alkoxy; and when R a is alkyl, Cy1 is a bridged bicyclic subcycloalkyl;

[0007] In some embodiments, X is CH 2 .

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

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

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

[0011] In some embodiments, R 1 and R 2 are each independently hydrogen or C 1-6 alkyl; 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.

[0012] In some embodiments, R a is -F, -Cl, -CF 3 , hydroxyalkyl, alkoxy or -CH 3 , provided that when R a is -CH 3 , Cy 1 is C 5 -C 10 bridged bicyclic subcycloalkyl.

[0013] In some other embodiments, the compound is of formula II: wherein Cy 1 is an arylene or a bridged bicyclic subcycloalkyl; and R a is halogen, alkyl, haloalkyl, hydroxyalkyl or alkoxy, provided that when R a is alkyl, Cy 1 is a bridged bicyclic subcycloalkyl.

[0014] In other embodiments, Cy 1 is phenylene or C 5 -C 10 bridged bicyclic subcycloalkyl. Examples of C 5 -C 10 bridged bicyclic subcycloalkyl include, but are not limited to

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

[0016] Another aspect of the present invention provides a pharmaceutical composition, wherein each pharmaceutical composition comprises the compound or a pharmaceutically acceptable salt thereof, and a pharmaceutically or physiologically acceptable carrier or excipient.

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

[0018] In some embodiments, the pharmaceutical composition is used in combination with a radiotherapy agent.

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

[0020] In some embodiments, the disease 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, allergies, inflammatory bowel disease, interstitial cystitis, mast cell activation syndrome, macrocythemia, otitis, pelvic inflammatory disease, reperfusion injury, rheumatic fever, rheumatoid arthritis, rhinitis, sarcoidosis and / or vasculitis.

[0022] Examples of cancers 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, gastric cancer, colorectal cancer, liver cancer, pancreatic cancer, skin cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, bone cancer, lymphoma and blood cancer.

[0023] The scope of the present invention also includes the use of the above compound in the preparation of a medicament for treating a subject suffering from a disorder mediated by the action of PGE2 on the EP4 receptor. Detailed Description

[0024] Reference will now be made in detail to the preferred embodiments of the present invention, and examples thereof will be further described. Although the present invention will be described in conjunction with the preferred embodiments, it should be understood that these preferred embodiments are not intended to limit the present invention to these embodiments. On the contrary, the present invention is intended to cover alternatives, modifications, and equivalents that may be included within the spirit and scope of the present invention as defined by the claims. In addition, in the specific embodiments, many specific details are set forth to provide a thorough understanding of the present invention. However, it will be apparent to those skilled in the art that the present 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 present invention. Definitions

[0025] As used herein, the term "or" is intended to include "and" and "or". In other words, the term "or" may also be replaced by "and / or".

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

[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 being, including a human patient or subject to whom the compositions of the present invention may be administered. The term "mammal" includes human patients and 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 containing at least one double bond or triple bond.

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

[0031] As used herein, the term "alkyl", by itself or as part of another substituent, refers to a straight-chain (i.e., unbranched) or branched hydrocarbon chain group composed of carbon and hydrogen atoms, which contains no unsaturation and has the stated number of carbon atoms (e.g., C 1 -C 10 or C 1 -C 10(alkyl). Whenever used herein, a numerical range (e.g., "1 to 10") refers to each integer within the given 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 and including 10 carbon atoms, although this definition also encompasses the occurrence of the term "alkyl" without a specified numerical range. Representative saturated linear or straight-chain alkyl groups include, but are not limited to, -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, and -n-hexyl; while saturated branched alkyl groups 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 group is attached to the parent molecule by a single bond. Unless otherwise specified in this specification, the alkyl group may optionally be substituted by 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 group as defined above, in which one or more hydrogen atoms have been replaced by a halogen, the halogen being independently selected from fluorine, chlorine, bromine, and iodine. "Fluoroalkyl" refers to an alkyl group as defined above, in which one or more hydrogen atoms have been replaced by fluorine atoms.

[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 group having at least one carbon-carbon double bond, which is obtained by removing a hydrogen atom from a single carbon atom of the parent alkene. The group can be in the cis or trans conformation of the double bond. Typical alkenyl groups include, but are not limited to, vinyl, propenyl, etc.

[0034] As used herein, the term "alkynyl" by itself or as part of another substituent refers to a carbon chain containing at least one carbon-carbon triple bond, which can be straight-chain, branched, or a combination thereof. Examples of alkynyl groups include ethynyl, propargyl, 3-methyl-1-pentynyl, 2-heptynyl, etc.

[0035] As used herein, the term "cycloalkyl", by itself or as part of another substituent, refers to the cyclic form of "alkyl". A cycloalkyl can include zero bridgehead carbon atoms or two or more bridgehead carbon atoms. Thus, a cycloalkyl can be monocyclic, bicyclic or polycyclic, depending on the number of bridgehead carbon atoms and bridging carbon atoms. A cycloalkyl including zero bridgehead carbon atoms is referred to herein as a "monocyclic cycloalkyl" or "unbridged cycloalkyl". A cycloalkyl including at least two bridgehead carbon atoms and at least one bridging carbon atom is referred to herein as a "bridged cycloalkyl". A bridged cycloalkyl including two bridgehead carbon atoms is referred to herein as a "bicyclic bridged cycloalkyl" or "bridged bicyclic cycloalkyl". A bridged cycloalkyl including 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" is a non-aromatic carbocyclic ring containing at least three carbon atoms and containing at least one carbon-carbon double bond, i.e., C═C. Examples of cycloalkenyl include, but are not limited to, cyclopropenyl, cyclobutenyl, cyclopentenyl, cyclopentadienyl, cyclohexenyl, cyclohexadienyl, etc. The term "heterocycloalkenyl" is a type of cycloalkenyl as defined above, in which at least one of the carbon atoms of the ring is replaced by a heteroatom, such as, but not limited to, nitrogen, oxygen, sulfur or phosphorus. Cycloalkenyl and heterocycloalkenyl can be substituted or unsubstituted.

[0037] As used herein, the term "heterocyclic group" 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 heterocyclic 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 "hydroxyl group" refers to the group -OH.

[0039] The groups defined above can include prefixes and / or suffixes commonly used in the art to produce other recognized substituents. For 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 an alkyl. Also, for example, "haloalkyloxy" or "haloalkyloxyl" refers to a group of the formula -OR', where R' is a haloalkyl.

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

[0041] As used herein, the term "aryl" refers to a group of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., sharing 6, 10, or 14 π electrons in a cyclic array), where the aromatic ring system has 6-14 ring carbon atoms and zero heteroatoms ("C 6-14 aryl"). In some embodiments, aryl has 6 ring carbon atoms ("C 6 aryl"; e.g., phenyl). In some embodiments, aryl has 10 ring carbon atoms ("C 10 aryl"; e.g., naphthyl, such as 1-naphthyl and 2-naphthyl). In some embodiments, aryl has 14 ring carbon atoms ("C 14 aryl"; e.g., anthryl).

[0042] As used herein, the term "heteroaryl" refers to a group of a monocyclic or polycyclic (e.g., bicyclic or tricyclic) 4n+2 aromatic ring system (e.g., sharing 6, 10, or 14 π electrons in a cyclic array), where the aromatic ring system has ring carbon atoms and one or more ring heteroatoms, and each heteroatom is independently selected from nitrogen, oxygen, and sulfur. In a heteroaryl containing one or more nitrogen atoms, the point of attachment can be a carbon atom or a nitrogen atom, as long as the valence allows. The heteroaryl polycyclic system can contain one or more heteroatoms in one or two rings.

[0043] The term "alkylene", by itself or as part of another substituent, refers to a bidentate moiety obtained by removing two hydrogen atoms from a hydrocarbon compound, where both hydrogen atoms are from the same carbon atom or one from each of two different carbon atoms, and the hydrocarbon compound can be aliphatic or cycloaliphatic and can be saturated, partially unsaturated, or fully unsaturated. Thus, the term "alkylene" includes subclasses such as 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 -, etc.) indicate the number of carbon atoms or the range of the number of carbon atoms. For example, the term "C 1-4 alkylene" as used herein refers to an alkylene having 1 to 4 carbon atoms.

[0044] Examples of straight-chain saturated C 1-8 alkylene include, but are not limited to, -(CH 2 ) n -, where n is an integer from 1 to 8, such as -CH 2 -, -CH 2 CH 2 CH 2 -, and -CH 2 CH 2 CH 2 CH2 -.

[0045] Branched saturated C 1-7 Examples of alkylene groups include, but are not limited to, -CH(CH 3 )-, -CH(CH 3 )CH 2 -, -CH(CH 3 )CH 2 CH 2 -, -CH(CH 3 )CH 2 CH 2 CH 2 -, -CH 2 CH(CH 3 )CH 2 -, -CH 2 CH(CH 3 )CH 2 CH 2 -, -CH(CH 2 CH 3 )-, -CH(CH 2 CH 3 )CH 2 - and -CH 2 CH(CH 2 CH 3 )CH 2 -.

[0046] Linear partially unsaturated C 1-7 Examples of alkylene groups include, but are not limited to, -CH=CH-, -CH=CH-CH 2 -, -CH=CH-CH 2 -CH 2 -, -CH=CH-CH 2 CH 2 CH 2 -, -CH=CH-CH=CH-, -CH=CH-CH=CH-CH 2 -, -CH=CH-CH=CH-CH 2 -CH 2 -, -CH=CH-CH 2 -CH=CH- and -CH=CH-CH 2 -CH 2 -CH=CH-.

[0047] Branched partially unsaturated C 1-7 Examples of alkylene groups include, but are not limited to, -C(CH 3 )=CH-, -C(CH 3 )=CH-CH 2- and -CH=CH-CH(CH 3 )-.

[0048] Alicyclic saturated C 1-7 Examples of the alkylene group include, but are not limited to, cyclopentylene (e.g., cyclopentane-1,3-diyl) and cyclohexylene (e.g., cyclohexane-1,4-diyl).

[0049] Alicyclic partially unsaturated C 1-7 Examples of the alkylene group include, but are not limited to, cyclopentenylene (e.g., 4-cyclopentene-1,3-diyl), cyclohexenylene (e.g., 2-cyclohexene-1,4-diyl; 3-cyclohexene-1,2-diyl; 2,5-cyclohexadiene-1,4-diyl).

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

[0051] As used herein, the term "absent" defines a variable, such as X, which means that the variable is absent, and thus the two groups connected by the variable are directly connected to each other. For example, in -N-X-Cy 2 when X is absent, Cy 2 and N are directly connected to each other.

[0052] The term "optional" or "optionally" means that the subsequent described event or situation may but does not necessarily occur, and the description includes the situation where the event or situation occurs and the situation where it does not occur. For example, "heterocyclic group optionally substituted by alkyl" means that the alkyl may or may not be present, and the description includes the situation where the heterocyclic group is substituted by alkyl and the situation where the heterocyclic group is not substituted by alkyl. Isomeric forms

[0053] The present invention provides novel compounds of formula I or pharmaceutically acceptable salts thereof, which, as EP4 receptor antagonists, can be used to treat PGE2-mediated diseases or disorders.

[0054] It should be understood that certain compounds (or salts, prodrugs or conjugates) of formula I may exist and be separable in isomeric forms, including tautomeric forms, cis- or trans-isomers, and optically active, racemic or diastereoisomeric forms. It should be understood that the present invention encompasses compounds of formula I in any tautomeric form or mixtures thereof; or mixtures of diastereoisomers, as well as individual diastereoisomeric forms, and the present invention encompasses compounds of formula I as mixtures of enantiomers, as well as individual enantiomeric forms, wherein any mixture or form has antagonist properties against the EP4 receptor. How to prepare or isolate a particular form and how to determine the antagonist properties against the EP4 receptor by standard tests (including the tests described below) are well known in the art.

[0055] In addition, the compounds of formula I (or their salts, prodrugs or conjugates) may exhibit polymorphism, or may form solvates with water or organic solvents. The present invention also encompasses any such polymorphism, any solvate or any mixture thereof.

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

[0057] Alternatively, any enantiomer of the compounds of formula I can be obtained by stereoselective synthesis using optically pure starting materials or reagents of known configuration. Salts

[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 salts are suitable, within the scope of reasonable medical judgment, for contact with the tissues of a patient without undue toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio, and effective for their intended use. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, iron, ferrous, lithium, magnesium, manganese salts, manganous, potassium, sodium, zinc, etc. Particularly preferred are ammonium salts, calcium salts, magnesium salts, potassium salts and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of the following: 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, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.

[0059] When the compounds of the present invention are basic, they can be prepared as salts with pharmaceutically acceptable non-toxic acids (including inorganic acids 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, hydroxyethanesulfonic 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 should be understood that the compounds of formula I mentioned herein also include pharmaceutically acceptable salts. Prodrug

[0061] As used herein, the term "prodrug" refers to an agent that is converted in vivo to the parent drug. Prodrugs are often useful because in some cases they may be more readily administered than the parent drug. For example, they may be bioavailable by oral administration while the parent drug is not. The solubility of prodrugs in pharmaceutical compositions is also improved compared to the parent drug. A non-limiting example of a prodrug is a compound of formula I that is administered as an ester ("prodrug") to facilitate transmembrane transport (where water solubility is disadvantageous for mobility), but once inside the cell, it is metabolically hydrolyzed to the carboxylic acid (the active entity), at which point water solubility is beneficial. Another example of a prodrug (likewise not intended to limit the scope of the term) could be a short peptide conjugated to an acid group that is converted to the active moiety inside the cell.

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

[0063] The compounds of the present invention are antagonists of the EP4 receptor and are thus expected to be useful for treating EP4 receptor-mediated diseases.

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

[0065] In another aspect, the present invention includes the use of a compound of formula I in the manufacture of a medicament for treating a disease or condition mediated by the action of PGE2 on the EP4 receptor.

[0066] As used herein, the term "treating a prostaglandin E2 (PGE2)-mediated disease or disorder" or "treating a disease or disorder mediated by the action of PGE2 on the EP4 receptor" refers to treating or preventing any chronic disease or disorder that can be beneficially treated or prevented by a selective EP4 antagonist. The term includes alleviating pain, fever, and inflammation of various conditions, including rheumatic fever, symptoms associated with influenza or other viral infections, the common cold, low back pain, neck pain, dysmenorrhea, headache, migraine, toothache, sprains and strains, myositis, neuralgia, synovitis, arthritis (including rheumatoid arthritis), degenerative joint disease (osteoarthritis), gout, ankylosing spondylitis, bursitis, burns, injuries, and pain and inflammation following surgery. In addition, this compound can inhibit cellular tumor transformation and metastatic tumor growth and thus can be used to treat and / or prevent cancer.

[0067] Examples of cancers include, but are not limited to, breast cancer, cancers that may be associated with Li-Fraumeni syndrome (such as childhood sarcoma, leukemia, and brain cancer), cancers that may be associated with Lynch syndrome (such as colon cancer), cholangiocarcinoma, brain cancer, endometrial cancer, kidney cancer, ovarian cancer, pancreatic cancer, small intestine cancer, stomach cancer, and ureteral cancer, lung cancer, melanoma, prostate cancer, retinoblastoma, thyroid cancer, and uterine cancer. Additionally, cancers may be the result of acquired mutations, such as those caused by diet, environment, and / or lifestyle, or somatic mutations. Examples of such cancers may include, but are not limited to, adrenal cancer, adrenocortical carcinoma, bladder cancer, brain cancer, 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 or 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 lymphoma, non-Hodgkin lymphoma), melanoma, malignant melanoma, malignant carcinoid, malignant pancreatic insulinoma, myeloma, multiple myeloma, ovarian cancer, pancreatic cancer (such as pancreatic exocrine cancer), prostate cancer, renal cell carcinoma, skin cancer (for example, in addition to the skin cancers mentioned previously, squamous cell carcinoma), stomach cancer, testicular cancer, thyroid cancer, thyroid follicular carcinoma, Wilms tumor, choriocarcinoma, mycosis fungoides, malignant hypercalcemia, cervical dysplasia, leukemia, acute lymphocytic leukemia, chronic lymphocytic leukemia, hairy cell lymphoma, Burkitt lymphoma, acute myeloid leukemia, chronic myeloid leukemia, myelodysplastic syndrome, promyelocytic leukemia, chronic granulocytic leukemia, acute granulocytic leukemia, fibrosarcoma, rhabdomyosarcoma, astrocytoma, neuroblastoma, rhabdomyosarcoma, schwannoma, Kaposi sarcoma, polycythemia vera, essential thrombocythemia, Hodgkin disease, non-Hodgkin lymphoma, soft tissue sarcoma, osteosarcoma, primary macroglobulinemia, seminoma, teratocarcinoma, osteosarcoma, xeroderma pigmentosum, keratoacanthoma, and retinoblastoma.

[0068] As used herein, the terms “treat,” “treatment,” or “treating,” etc., refer to any indication of success in preventing or ameliorating an injury, pathology, or disorder, including any objective or subjective parameter, such as alleviation of symptoms; remission; elimination or making the injury, pathology, or disorder more tolerable to the patient; slowing the rate of degeneration or decline; making the ultimate point of degeneration less debilitating; or improving the physical or mental health of the subject. Treatment or amelioration of symptoms can be based on objective or subjective parameters, including the results of a physical examination, neurological examination, and / or psychiatric evaluation.

[0069] "Effective amount" or "therapeutically effective amount" refers to an amount found to be effective in treating prostaglandin E2-mediated diseases or disorders or cancer by methods such as clinical testing and evaluation, patient observation, etc. "Effective amount" can also refer to an amount that results in a detectable change in biological or chemical activity. A person skilled in the art can detect and / or further quantify the detectable change for the relevant mechanism or process. In addition, "effective amount" can refer to an amount that maintains the desired physiological state, i.e., an amount that reduces or prevents significant decline and / or promotes improvement of the condition. "Effective amount" can also refer to an amount that is therapeutically effective.

[0070] The compound of Formula I can also be used in combination with radiation and / or one or more therapeutic agents selected from chemotherapeutic agents, anti-cytotoxic T lymphocyte antigen 4 antibody (anti-CTLA4), anti-programmed death ligand 1 antibody (anti-PDL1), anti-programmed cell death protein 1 antibody (anti-PD1), indoleamine-2,3-dioxygenase (IDO) inhibitor, tryptophan-2,3-dioxygenase (TDO) inhibitor, 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 ( Merck).

[0071] Examples of chemotherapeutic agents include, but are not limited to, aromatase inhibitors; antiestrogens, antiandrogens (especially in the case of prostate cancer), or gonadotropin-releasing hormone agonists; topoisomerase I inhibitors or topoisomerase II inhibitors; microtubule active agents, alkylating agents, antitumor antimetabolites, or platinum compounds; compounds that target / reduce protein or lipid kinase activity or protein or lipid phosphatase activity, further antiangiogenic compounds, or compounds that induce the process of cell differentiation; bradykinin I receptor or angiotensin II antagonists; cyclooxygenase inhibitors, bisphosphonates, rapamycin derivatives (such as everolimus), heparinase inhibitors (preventing heparan sulfate degradation) such as PI-88, biologic response modifiers (preferably lymphokines or interferons, such as interferons), ubiquitination inhibitors, or inhibitors that block the anti-apoptotic pathway; inhibitors of Ras oncogenic isoforms, such as H-Ras, K-Ras, or N-Ras, or farnesyl transferase inhibitors, such as L-744,832 or DK8G557; telomerase inhibitors, such as telomestatin; protease inhibitors, matrix metalloproteinase inhibitors, methionine aminopeptidase inhibitors, such as benzamide or its derivatives, or proteasome inhibitors, such as PS-341; histone deacetylase inhibitors, such as vorinostat, MG0103, or MS275; and kinase inhibitors.

[0072] The pharmaceutical composition of the present invention comprises a compound of formula I or a pharmaceutically acceptable salt thereof as an active ingredient, and may further comprise a pharmaceutically acceptable carrier and optionally other therapeutic ingredients.

[0073] The EP4 antagonist, antibody, and / or antimetabolite can be administered to a subject by any suitable route, including orally (including buccal administration, also including oral gavage), intraperitoneally, parenterally, by inhalation spray, topically (i.e., on skin and mucosal surfaces, including airway surfaces), transdermally, rectally, nasally (including nasogastric tube), sublingually, orally, vaginally, or by an implantable reservoir. As used herein, the term "parenterally" includes subcutaneous, intramuscular, intradermal, intravenous, intra-articular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. In a specific embodiment, 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] A pharmaceutical composition containing an active ingredient may be in a form suitable for oral use, such as tablets, lozenges, troches, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs. The compositions for oral use can be prepared by any method known in the art of pharmaceutical composition manufacture, and the compositions may contain one or more agents selected from sweetening agents, flavoring agents, coloring agents, and preservatives to provide a pharmaceutically elegant and palatable preparation. Tablets contain a mixture of the active ingredient and a pharmaceutically acceptable non-toxic excipient suitable for the preparation of tablets. These excipients can be, for example, 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; binding agents such as starch, gelatin or gum arabic; and lubricants such as magnesium stearate, stearic acid or talc. Tablets may be uncoated or 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 delayed release material such as glyceryl monostearate or glyceryl distearate may be used. They 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 invention and are not meant to limit the scope of the invention. Examples Assays for determining biological activity

[0076] The compounds of formula I can be tested using the following assays to determine their in vitro and in vivo prostaglandin antagonist or agonist activity and their selectivity. The prostaglandin receptor activities described are for DP, EP 1 、EP 2 、EP 3 、EP 4 、FP, IP and TP. Example A. Stable expression of prostaglandin receptors in the human embryonic kidney (HEK) 293 (ebna) cell line

[0077] The prostaglandin receptor cDNA corresponding to the full-length coding sequence was subcloned into the appropriate site of a mammalian expression vector and transfected into HEK293(ebna) cells. HEK293(ebna) cells expressing a single cDNA were cultured under selective conditions and single colonies were isolated using a cloning ring-based method after 2 - 3 weeks of growth, and subsequently expanded into clonal cell lines. Example B. Prostaglandin receptor binding assay

[0078] The transfected HEK293 (ebna) cells were maintained in culture medium and harvested. After lysing the cells in the presence of protease inhibitors, membranes were prepared by differential centrifugation for receptor binding assays. Prostaglandin receptor binding assays (for DP1, DP2 (CRTH2), EP1, EP2, EP3-III, EP4, FP, IP, and TP) were performed in 10 mM MES / KOH (pH 6.0) (for EP, FP, and TP) or 10 mM HEPES / KOH (pH 7.4) (for DP and IP), which contained 1 mM EDTA, 2.5 - 30 mM divalent cations, and an appropriate radioligand that binds specifically to the target prostaglandin receptor. The synthetic compound was added to dimethyl sulfoxide, which was kept constant at 1% (v / v) during all incubations. The reaction was initiated by adding membrane proteins. Nonspecific binding was determined in the presence of 10 μM of the corresponding non-radiolabeled prostaglandin. Incubation was carried out at room temperature or 30 °C for 60 - 90 minutes and terminated by rapid filtration. After subtracting nonspecific binding from total binding, compound binding was calculated as the percentage inhibition of radioligand binding. The residual specific binding at each ligand concentration was calculated and expressed as a function of ligand concentration to construct a sigmoidal concentration-response curve. The Hill equation curve fitting formula Y = D + [(A - D) / (1 + C / IC50 H was used to perform nonlinear regression analysis on the concentration-response curve to determine the half-maximal inhibitory concentration IC 50 value of the compound, where Y = specific binding, A = left asymptote of the curve, D = right asymptote of the curve, C = compound concentration, and H = slope factor. The binding affinity of the compound was determined by calculating the equilibrium inhibition constant (K i ) from the equation K i = InPt / 1 + [radioligand] / Kd, where K d 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 conditions: Table 1. Inhibition of PGE2 binding by representative compounds Example C. Microsomal Stability Test

[0080] Microsomal stability tests were conducted to understand the metabolism of representative compounds. In vitro microsomal stability was determined in pooled human, dog, rat, and mouse liver microsomes. The test compounds (Table 2) were incubated for 0 - 60 minutes in the presence and absence of NADPH, and the amount of remaining compound was quantified by LC-MS / MS analysis. Table 2. Hepatic microsomal stability of representative compounds Example D. Pharmacokinetic (PK) tests The test compounds were dissolved in 10% DMSO, 40% PEG-400, and 50% water for intravenous (i.v.) and oral (p.o.) administration to rats. Three male Sprague-Dawley rats received a single i.v. dose (2 mg / kg) via the tail vein. Three male animals received a single p.o. dose (5 mg / kg) via a feeding 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 after dosing. 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. Table 3. Rat PK test of Example 1 Example E. Prostanoid receptor agonist and antagonist tests

[0081] Whole cell second messenger tests were performed to measure the stimulation of intracellular cAMP accumulation in HEK-293(ebna)-hEP4 cells to determine whether the receptor ligand is an agonist or an antagonist. Cells were harvested and resuspended in HBSS containing 25 mM HEPES, pH 7.4. The incubation medium contained 0.5 mM IBMX (a phosphodiesterase inhibitor available from Biomol). Samples were incubated at 37 °C for 10 minutes, the reaction was terminated, and then the cAMP level was measured. The ligand was added to dimethyl sulfoxide and kept constant at 1% (v / v; agonist) or 2% (v / v; antagonist) in all incubation media. For agonists, the second messenger response was expressed as a function of ligand concentration, and the EC 50 value and the maximal response were calculated compared to the PGE2 standard. For antagonists, the ability of the ligand to inhibit the agonist response was determined by performing a dose-response curve at a concentration corresponding to its EC 2 in the presence of the PGE2 70 agonist. The IC 50 value was calculated as the ligand concentration required to inhibit 50% of the PGE2-induced activity. Example F. Rat Paw Edema Test

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

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

[0084] Female Lewis rats (weighing approximately 146 - 170 g) were weighed, ear - tagged, and grouped (negative control group without induced arthritis, vehicle control group, positive control group given indomethacin at a total daily dose of 1 mg / kg, and four groups given the test compound at total daily doses of 0.10 - 3.0 mg / kg) such that the body weights within each group were equal. Each of 10 rats in 6 groups was injected with 0.5 mg Mycobacterium butyricum (adjuvant) in 0.1 mL light mineral oil into the hind paws; 10 rats in the negative control group were not injected with adjuvant. Body weight, contralateral paw volume (measured by mercury displacement plethysmography), and lateral X - ray films (obtained under ketamine and xylazine anesthesia) were measured before injection of adjuvant (day - 1) and 21 days after injection of adjuvant. The volume of the main paw was measured before injection of adjuvant (day - 1) and on days 4 and 21 after injection of adjuvant. Rats were anesthetized by intramuscular injection of a mixture of 0.03 - 0.1 mL of ketamine (87 mg / kg) and xylazine (13 mg / kg) for taking X - ray films and injecting adjuvant. X - ray films of the double hind paws were taken using a Faxitron (45 kVp, 30 s) and Kodak X - OMAT TL film on days 0 and 21 and developed in an automatic processor. Researchers blinded to the experimental treatment evaluated the changes in soft and hard tissues in the X - ray films. The following X - ray film changes were numerically graded according to severity: increased soft - tissue volume (0 - 4), narrowing or widening of the joint space (0 - 5), subchondral erosion (0 - 3), periosteal reaction (0 - 4), osteolysis (0 - 4), subluxation (0 - 3), and degenerative joint changes (0 - 3). Specific criteria were used to determine the numerical grade of severity for each X - ray film change. The maximum possible score for each foot was 26. Starting from the day after injection of adjuvant, the test compound at total daily doses of 0.1, 0.3, 1, and 3 mg / kg / day, indomethacin at a total daily dose of 1 mg / kg / day, or vehicle (0.5% Methocel TMSterile water) was administered orally twice a day for 21 days. The compound was prepared once a week, stored refrigerated in the dark before use, and vortexed immediately before administration. Example I. Syngeneic mouse model for measuring anti-tumor activity

[0085] The assays described in the article by Spranger et al. (Journal for ImmunoTherapy of Cancer, 2014, 2:3) can be used to evaluate the synergistic effects of the compounds of the present invention in combination with an effective amount of anti-cytotoxic T lymphocyte antigen 4 antibody (anti-CTLA4); anti-programmed death ligand 1 antibody (anti-PDL1); anti-programmed cell death protein 1 antibody (anti-PD1); indoleamine-2,3-dioxygenase (IDO) inhibitor; tryptophan-2,3-dioxygenase (TDO) inhibitor. Synthesis of the compound Example 1: 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoic acid Route 1 Step 1: 4-bromo-5-chloro-thiophene-2-carbaldehyde

[0086] Under argon at -78 °C, LDA (2.5 M, 15.19 mL, 1.5 equivalents) was added dropwise over 45 minutes to a solution of 3-bromo-2-chloro-thiophene (5 g, 25.32 mmol, 1 equivalent) in THF (50 mL). The reaction mixture was stirred at -78 °C for 1 hour (h), then anhydrous DMF (9.25 g, 126.59 mmol, 9.74 mL, 5 equivalents) was added at -78 °C over 15 minutes. After addition was complete, the temperature was raised to 25 °C and stirred for 45 minutes. The reaction was monitored by TLC. After completion of the reaction, aqueous citric acid solution was added and the reaction mixture was stirred for 5 minutes. 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 (SiO 2 , petroleum ether / ethyl acetate = 5 / 1). The compound 4-bromo-5-chloro-thiophene-2-carbaldehyde (4.7 g, 20.84 mmol, yield 82.3%) was obtained as a yellow solid. Step 2: Methyl (E)-2-azido-3-(4-bromo-5-chloro-2-thienyl)prop-2-enoate

[0087] At 0 °C, a solution of NaOMe in MeOH (4.36 M, 18.31 mL, 1 equiv) was added to a solution of 4-bromo-5-chloro-thiophene-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). After the addition was complete, the mixture was stirred at 0 °C for 2 h. The reaction was monitored by TLC. After the reaction was complete, saturated NH 4 Cl was added at 25 °C to quench the reaction, then diluted with water, extracted with ethyl acetate (50 mL × 3), the organic layer was collected and dried over Na 2 SO 4 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

[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 2 three times, then the mixture was stirred at 140 °C for 4 h under a N 2 atmosphere. The reaction was monitored by TLC. After the reaction was completed, the solution was cooled to room temperature, allowed to stand overnight, the solid was collected by filtration and dried in vacuo 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 reaction without further purification. Step 4: Methyl 3-bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylate

[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) and 1-(bromomethyl)-4-(trifluoromethyl)benzene (1.15 g, 4.82 mmol, 743.45 uL, 1 equiv) in DMF (15 mL) was added Cs 2 CO 3(4.71 g, 14.46 mmol, 3 eq). The mixture was stirred at 25 °C for 3 h. The reaction was monitored by TLC. After 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 Na 2 SO 4 , filtered and concentrated under reduced pressure. The residue was purified by column chromatography (SiO 2 , 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 pos): 453.9 (M+2) + . Step 5: 3-Bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylic acid

[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 eq) in THF (12.5 mL) and MeOH (12.5 mL) was added LiOH·H 2 O (1 M, 6.63 mL, 3 eq) (1 N aqueous solution). The reaction mixture was stirred at 55 °C for 4 h. The reaction was monitored by LCMS. After completion of the reaction, most of the solvent was removed by concentration. The residue was diluted with water and acidified to pH = 3 - 4 with 1 N HCl and extracted with EtOAc (15 mL × 3). The combined organic layers were dried over Na 2 SO 4 , 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 pos): 439.0 (M+2) + . Step 6: 3-Bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole

[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 eq) in DMSO (8 mL) was added Ag2 CO 3 (502.90 mg, 1.82 mmol, 82.71 uL, 1 equiv) and 2 - 3 drops of HOAc. The mixture was stirred at 120 °C under N 2 for 16 h. The reaction was monitored by LCMS. After completion of the reaction, the reaction mixture was diluted with H 2 O (15 mL) and extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 and 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, yield 90.31%) as a light yellow solid, which was used in the next step without further purification. MS (ES - API pos): 396.0 (M + 2) + . Step 7: 2 - chloro - 4 - [[4 - (trifluoromethyl)phenyl]methyl]thieno[3,2 - b]pyrrole - 3 - carboxylic acid

[0092] At - 78 °C and under N 2 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 complex (1.3 M, 1.27 mL, 1.3 equiv). The reaction mixture was stirred at - 78 °C for 30 min. Then, dry CO 2 was bubbled into the reaction mixture at - 78 °C for 20 min. The reaction was monitored by LCMS. After completion of the reaction, the reaction was quenched by adding water (8 mL), and then extracted with EtOAc (10 mL × 3). The organic layer was dried over Na 2 SO 4 and 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, yield 60.55%, purity 60%) was obtained as a light yellow oil, which was used directly in the next step without further purification. MS (ES - API pos): 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

[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. Then methyl 4-(1-aminocyclopropyl)benzoate (2.84 g, 14.87 mmol, 1 equiv) was added to the reaction mixture, and then 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 and 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 pos): 533.1 (M+1) + 。 1 1H-NMR (500 MHz, DMSO-d6) δ 9.33 (s, 1H), 7.73 (d, J = 6.4 Hz, 2H), 7.64 (br d, J = 6.4 Hz, 2H), 7.27 - 7.20 (m, 3H), 7.07 (br d, J = 6.4 Hz, 2H), 6.50 (d, J = 2.4 Hz, 1H), 5.45 (s, 2H), 3.82 (s, 3H), 1.26 - 1.20 (m, 2H), 0.99 - 0.93 (m, 2H). Step 9: 4-[1-[[2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoic acid

[0094] To 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 H 2To a mixture of O (40 mL) was added LiOH·H 2 O (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 to pH ~ 4 with 1 N HCl. Then it was extracted with EtOAc (200 mL × 3), and the combined organic layers were dried over Na 2 SO 4 dried, 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, yield 76.52%, purity 100%) as a pale yellow solid. 1 1H-NMR (400 MHz, DMSO-d6) δ 12.80 (br s, 1H), 9.33 (s, 1H), 7.74 (d, J = 6.4 Hz, 2H), 7.63 (d, J = 6.4 Hz, 2H), 7.30 - 7.18 (m, 3H), 7.07 (d, J = 6.4 Hz, 2H), 6.50 (d, J = 2.4 Hz, 1H), 5.45 (s, 2H) 1.25 - 1.18 (m, 2H), 0.96 - 0.90 (m, 2H). MS (ES-API pos): 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

[0095] At 20 °C, to 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), H 2To a solution of O (100 mL) and MeOH (50 mL) was added NaOH (5.85 g, 146.35 mmol, 3 equiv), and then the mixture was 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 h. The solid was collected by filtration, and the filter cake was washed with water (100 mL × 2) and dried in vacuo to obtain 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%), which was a pale yellow solid. 1 1H-NMR (400 MHz, DMSO-d6) δ 7.68 (d, J = 6.8 Hz, 2H), 7.62 (d, J = 6.4 Hz, 2H), 7.27 (d, J = 6.8 Hz, 2H), 7.08 (d, J = 2.4 Hz, 1H), 6.98 (d, J = 2.4 Hz, 2H), 6.43 (d, J = 2.4 Hz, 1H), 5.41 (s, 2H), 1.18 (t, J = 7.2 Hz, 2H), 0.93 (t, J = 7.2 Hz, 2H). MS (ES-API pos): 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 Route 2 Step 1: 3-Bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-5-carboxylic acid

[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. After completion of the reaction, most of the solvent was removed by concentration, then diluted with water and acidified to pH = 3 - 4 with 1 N HCl, and extracted with EtOAc (15 mL × 3). The combined organic layers were washed with Na 2 SO 4Dry, filter and concentrate 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, yield 98.04%) as a white solid. MS (ES-API pos): 620.4 (M+1) + 。 Step 2: 3-Bromo-2-chloro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole

[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 Ag 2 CO 3 (502.90 mg, 1.82 mmol, 82.71 μL, 1 equiv) and 50 mg HOAc. The mixture was stirred at 120 °C under N 2 for 16 h. The reaction was monitored by LCMS and after completion, diluted with H 2 O (50 mL), extracted with ethyl acetate (20 mL × 3). The combined organic layers were washed with brine, dried over Na 2 SO 4 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, yield 90.31%) as a pale yellow solid, which was used in the next step without further purification. MS (ES-API pos): 396.0 (M+1) + 。 Step 3: 4-[[4-(trifluoromethyl)phenyl]methyl]-2-(triisopropylsilyloxymethyl)thieno[3,2-b]pyrrole-3-carboxylic acid

[0098] Under N 2 at -78 °C, isopropylmagnesium chloride lithium chloride complex (1.3 M, 985.19 μL, 2 equiv) was added to a solution of [3-bromo-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole-2-yl]methoxy-triisopropyl-silane (350 mg, 640.37 μmol, 1 equiv) in THF (7 mL) over 5 min. After addition was complete, the reaction mixture was warmed to -10 °C and stirred at -10 °C for 30 min (min), then cooled back to -78 °C, and then dry CO was added to the reaction mixture at -78 °C2 Bubble for 20 minutes. Monitor the reaction by LCMS. After the reaction is completed, quench the reaction by adding water and extract with EtOAc (10 mL × 3). The organic layer is dried over Na 2 SO 4 dry, filter and concentrate in vacuo to give 4-[[4-(trifluoromethyl)phenyl]methyl]-2-(triisopropylsilyloxymethyl)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

[0099] To a solution of 4-[[4-(trifluoromethyl)phenyl]methyl]-2-(triisopropylsilyloxymethyl)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. Then methyl 4-(1-aminocyclopropyl)benzoate (78.48 mg, 410.42 μmol, 1 equiv) was added to the reaction mixture and stirred at 25 °C for 12 hours. Monitor the reaction by LCMS. After the reaction is completed, the reaction mixture was diluted with EtOAc (60 mL), washed with brine (8 mL × 5), dried over Na 2 SO 4 dry, and then concentrated in vacuo to give methyl 4-[1-[[4-[[4-(trifluoromethyl)phenyl]methyl]-2-(triisopropylsilyloxymethyl)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

[0100] To a solution of methyl 4-[1-[[4-[[4-(trifluoromethyl)phenyl]methyl]-2-(triisopropylsilyloxymethyl)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 consumption of the starting material; the desired compound was observed. The reaction mixture was concentrated in vacuo. The residue was purified by column chromatography (SiO 2 , 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 pos): 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

[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 H 2 O (1 mL) was added LiOH·H 2 O (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. After completion of the reaction, the reaction mixture was acidified to pH ~4 with 1 N HCl and extracted with EtOAc (12 mL × 3). The organic layer was washed with brine (6 mL) and dried over Na 2 SO 4Dry, filter and concentrate. 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), then lyophilized 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, yield 37.65%), as a white solid. 1 1H NMR (500 MHz, DMSO-d6) δ 12.98 - 12.59 (m, 1H), 9.06 - 8.93 (m, 1H), 7.75 - 7.66 (m, 2H), 7.64 - 7.56 (m, 2H), 7.21 - 7.10 (m, 3H), 7.07 - 6.98 (m, 2H), 6.52 - 6.42 (m, 1H), 5.57 - 5.50 (m, 2H), 4.77 - 4.68 (m, 2H), 1.28 - 1.15 (m, 2H), 1.09 - 0.99 (m, 2H). 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) Route 3 Step 1: 4-Bromo-5-fluorothiophene-2-carbaldehyde

[0102] To a solution of 4-bromothiophene-2-carbaldehyde (7 g, 36.64 mmol, 1 equiv) in CH 3 CN (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. After completion of the reaction, most of the solvent was removed by concentration. The residue was diluted with water and extracted with EtOAc (3×100 mL). The combined organic layers were washed with brine and dried over Na 2 SO 4 dried, filtered and concentrated under reduced pressure. The residue was purified by flash silica gel chromatography ( 80g The silica gel flash column was eluted with a gradient of 0 - 20% ethyl acetate / petroleum ether (@20 mL / min) for purification to obtain 4-bromo-5-fluoro-thiophene-2-carboxaldehyde (5 g, 23.92 mmol, yield 65.28%), which was a white solid. Step 2: Methyl (E)-2-azido-3-(4-bromo-5-fluorothiophen-2-yl)acrylate

[0103] To a solution of 4-bromo-5-fluoro-thiophene-2-carboxaldehyde (4 g, 19.14 mmol, 1 equiv), methyl 2-azidoacetate (6.61 g, 57.41 mmol, 3 equiv) in MeOH (10 mL) was added NaOMe (4.36 M, 4.39 mL, 1 equiv). The mixture was stirred at 0 °C for 2 h. The reaction was monitored by LCMS. After completion of the reaction, the salts were filtered off and the filtrate was concentrated under reduced pressure to obtain crude methyl (E)-2-azido-3-(4-bromo-5-fluoro-thiophen-2-yl)prop-2-enoate (5 g, 16.33 mmol, yield 85.36%), which was a yellow solid and 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

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

[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 Cs 2 CO 3(1.41 g, 4.32 mmol, 3 eq). The mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, it was diluted with water and extracted with EtOAc (30 mL × 2). The combined organic layers were washed with brine and dried over Na 2 SO 4 dried, 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, yield 95.63%), 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

[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 eq) in MeOH (4 mL), THF (4 mL), H 2 O (2 mL) was added LiOH (329.42 mg, 13.75 mmol, 10 eq). The mixture was stirred at 25 °C for 16 h. The reaction was monitored by TLC. After completion of the reaction, it was diluted with water and extracted with EtOAc (15 mL × 3). The combined organic layers were washed with brine and dried over Na 2 SO 4 dried, 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, yield 34.44%), as a white solid. Step 6: 3-Bromo-2-fluoro-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole

[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 eq) in DMSO (1.5 mL) was added Ag 2 CO 3 (32.97 mg, 119.55 μmol, 5.42 μL, 1 eq) and CH 3COOH (717.93 μg, 11.96 μmol, 0.1 eq). The mixture was then stirred at 120 °C for 16 h. The reaction was monitored by LCMS. After completion of the reaction, it was diluted with water and extracted with EtOAc (15 mL × 2). The combined organic layers were washed with brine and dried over Na 2 SO 4 dried, 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

[0108] At -78 °C and under N 2 2, i-PrMgCl·LiCl (1.3 M, 370.19 μL, 1.3 eq) was added to a mixture of 3-bromo-2-fluoro-4-[[4-(trifluoromethyl)phenyl]methyl]thieno[3,2-b]pyrrole (140 mg, 370.19 μmol, 1 eq) in THF (3 mL). The reaction mixture was then stirred at -10 °C for 20 min, then cooled back to -78 °C and bubbled with dry CO 2 2 for 20 min. The reaction was monitored by LCMS. After completion of the reaction, the reaction was quenched by adding water and then extracted with EtOAc (10 mL × 3). The organic layer was dried over Na 2 SO 4 dried, 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

[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) was added HATU (172.78 mg, 454.42 μmol, 1.3 equiv) and TEA (70.74 mg, 699.11 μmol, 97.31 μL, 2 equiv). The reaction was carried out for 30 minutes, and then methyl 4-(1-aminocyclopropyl)benzoate (73.53 mg, 384.51 μmol, 1.1 equiv) was added. The mixture was stirred at 25 °C for 12.5 hours. The reaction was monitored by LCMS. After completion of the reaction, the reaction mixture was concentrated under reduced pressure. Then it was diluted with water and extracted with EtOAc (15 mL × 2). The combined organic layers were washed with brine, dried over 2 SO 4 Na, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (SiO 2 , 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, 22.15% yield) 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

[0110] To a solution of methyl 4-[1-[[2-fluoro-4-[[4-(trifluoromethyl)phenyl]methylthieno[3,2-b]pyrrole-3-carbonyl]amino]cyclopropyl]benzoate (30 mg, 58.08 μmol, 1 equiv) in THF (1 mL) and H 2To the solution in O (0.3 mL), LiOH (13.91 mg, 580.83 μmol, 10 eq) was added. The mixture was stirred at 40 °C for 12 h. The reaction was monitored by LCMS. After completion of the reaction, the solvent was removed by concentration, and the residue was purified by preparative HPLC (TFA conditions). Column: BostonGreen ODS 150×30 mm×5 μm; Mobile phase: [water (TFA)-can]; B%: 53% - 73%, 10 min), then lyophilized to obtain 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, yield 68.53%) 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, yield 50.2%). 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), 7.17–7.16 (d, J = 8.5 Hz, 2H), 7.07–7.05 (d, J = 8.0 Hz, 2H), 6.48 (s, 1H), 5.56 (s, 2H), 1.26 - 1.24 (m, 2H), 1.03 - 0.97 (m, 2H); MS (ES-API pos): 503.1 (M+1) + 。 1 H NMR (500 MHz, CD 3 Cl) 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 pos): 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 Route 4 Step 1: Benzyl 3-cyanobicyclo[1.1.1]pentane-1-carboxylate

[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 K 2 CO 3 (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 (r.t.) for 16 h. The reaction was monitored by TLC. After 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 Na 2 SO 4 , filtered, the benzyl bromide was removed and the residue was concentrated under reduced pressure; the residue was purified by silica gel column chromatography (gradient of EtOAc in petroleum ether from 0 to 5%) to afford 390 mg of the title product as a colorless oil. MS (ES-API pos): 227.1 (M+1) + . Step 2: Benzyl 3-(1-aminocyclopropyl)bicyclo[1.1.1]pentane-1-carboxylate

[0112] Under N 2 atmosphere at room temperature, to a solution of benzyl 3-cyanobicyclo[1.1.1]pentane-1-carboxylate (390 mg, 1.72 mmol) in Et 2 O (10 mL) was added Ti(Oi-Pr) 4 (536.52 mg, 1.89 mmol, 557.13 μL), 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 3 ·Et 2 O (487.13 mg, 3.43 mmol, 423.59 μL) was added dropwise at -78 °C. Then the reaction was slowly warmed to room temperature (25 °C) and stirred at 25 °C for 16 h under N 2 atmosphere. The reaction was monitored by LCMS. After the addition was complete, the reaction mixture was quenched dropwise with 1 M HCl at 25 °C and with H 2Dilute, then add 10% aqueous NaOH solution (15 mL) dropwise at 25 °C until pH = 7. After stirring at room temperature for 20 minutes, extract with EtOAc (20 mL × 2). The combined organic layers are dried over Na 2 SO 4 and concentrated under reduced pressure to obtain 300 mg of the title product as a yellow oil, which is directly used 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

[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) at 25 °C, add HATU (475.62 mg, 1.25 mmol) and DIEA (80.83 mg, 625.43 μmol, 108.94 μL). Stir the mixture at 25 °C for 1 hour. Monitor the reaction by LCMS. After the reaction is complete, dilute the reaction mixture with water and extract with EtOAc (15 mL × 3). The combined organic layers are washed with brine and then concentrated under reduced pressure to obtain 200 mg of the title product as a brown oil, which is directly used 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

[0114] To 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 H 2LiOH (79.95 mg, 3.34 mmol) was added to the solution in O (2.5 mL). The mixture was stirred at 55 °C for 2 h. The reaction was monitored by LCMS. After completion of the reaction, the reaction mixture was quenched with 1 M HCl until pH = 4 - 5, and then extracted with EtOAc (15 mL, 3 times). The combined organic layers were 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% - 72%, 10 min) to obtain 34.46 mg of the title product as a white solid. 1 H NMR (CD 3 OD, 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 pos): 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 Route 5 Step 1: Methyl 4-carbamoyl bicyclo[2.2.2]octane-1-carboxylate

[0115] A mixture of 4-methoxycarbonyl bicyclo[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 NH 4 Cl (25.20 g, 471.16 mmol) in DMF (100 mL) was stirred at 25 °C for 16 h. The reaction was monitored by LCMS. After completion of the reaction, the reaction mixture was diluted with water (about 50 mL) and extracted with EtOAc (50 mL×3). The combined organic layers were washed with brine and dried over Na 2 SO 4Dry, filter and concentrate under reduced pressure to obtain 9.9 g of the crude title compound as a pale yellow oil, which can be directly used in the next step. MS(ES-API positive): 212.3 (M+1) + 。 Step 2: Methyl 4-cyanobicyclo[2.2.2]octane-1-carboxylate

[0116] At 0 °C, TFAA (68.90 g, 328.04 mmol, 45.63 mL) was added dropwise to a solution of methyl 4-carbamoyl-bicyclo[2.2.2]octane-1-carboxylate (9.9 g, 46.86 mmol) in pyridine (80 mL). Then the mixture was slowly warmed to room temperature and stirred overnight. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was diluted with 1N HCl and extracted with EtOAc (30 mL × 3). The combined organic layers were washed with brine and dried over Na 2 SO 4 dried, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (EtOAc gradient in petroleum ether from 0 to 20%) to obtain 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

[0117] Under N 2 atmosphere at 25 °C, Ti(Oi-Pr) 2 (760.38 mg, 2.68 mmol, 789.59 μL) was added to a solution of methyl 4-cyanobicyclo[2.2.2]octane-1-carboxylate (470 mg, 2.43 mmol) in Et 4 O (30 mL), 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 hour, then BF 3 ·Et 2 O (690.40 mg, 4.86 mmol, 600.35 μL) was added dropwise at -78 °C, then the temperature was slowly raised to room temperature and under N 2Stir for 5 hours at 25 °C under an atmosphere. Monitor the reaction by LCMS. After the reaction is complete, quench the reaction by dropwise addition of 1 M HCl at room temperature, then dilute with water, and then add 10% aqueous NaOH solution (15 mL) dropwise at 25 °C until pH = 7. After stirring for 20 minutes at room temperature, extract with EtOAc (20 mL × 2). The combined organic layers are dried over Na 2 SO 4 and concentrated under reduced pressure to give 350 mg of the title product as a yellow oil, which can be 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

[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 hours. Monitor the reaction by LCMS. After the reaction is complete, dilute the reaction mixture with H 2 2O and extract 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 can be directly used 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

[0119] To 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 H2 To the solution in O (2.5 mL) was added LiOH (63.58 mg, 2.65 mmol). The mixture was stirred at 55 °C for 3 hours, and the reaction was monitored by LCMS. After the reaction was completed, the reaction mixture was diluted with H 2 O and 1 M HCl until pH = 4 - 5, and then extracted with EtOAc (15 mL × 3). The combined organic layers were 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% - 90%, 10 minutes) to obtain 45.48 mg of the title product as a white solid. 1 H NMR (CD 3 OD, 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 Route 6 Step 1: 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

[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). The mixture was stirred at 25 °C for 1 h. The reaction was monitored by LCMS. After 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

[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 H 2 O (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. After completion of the reaction, the reaction mixture was diluted with water and then 1 M HCl was added until pH = 4 - 5, and then it was extracted with EtOAc (15 mL × 3). The combined organic layers were 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% - 70%, 10 min) to give 55.14 mg of the title product as a white solid. 1 H NMR(CD 3OD, 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 Route 7 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

[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, and the reaction was monitored by LCMS. After completion of the reaction, 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

[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 H 2 O (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. After completion of the reaction, the reaction mixture was diluted with H 2 O (10 mL), then 1 M HCl was added until pH = 4 - 5, and then the mixture was extracted with EtOAc (15 mL × 3). The combined organic layers were 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% - 76%, 10 min) to give 14.59 mg of the title product as a white solid. 1 H NMR (CD 3 OD, 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 pos): 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 Route 8 Step 1: Methyl 3-[methoxy(methyl)carbamoyl]bicyclo[1.1.1]pentane-1-carboxylate

[0124] To 3-methoxycarbonylbicyclo[1.1.1]pentane-1-carboxylic acid (1.0 g, 5.88 mmol, 1 equiv), N,O-dimethylhydroxylamine hydrochloride (N-methoxymethanamine; hydrochloride, 687.89 mg, 7.05 mmol, 1.2 equiv) and Et3 A solution of N (1.78 g, 17.63 mmol, 2.45 mL, 3.0 equiv) in DCM (15 mL) was added to HATU (2.68 g, 7.05 mmol, 1.2 equiv). The mixture was stirred overnight at room temperature. The reaction was monitored by LCMS. After completion of the reaction, the reaction was quenched with water and then extracted with DCM (10 mL × 3). The combined organic layers were concentrated under reduced pressure. The residue was purified by flash silica chromatography ( 12 g Flash silica column, eluent with 0 - 25% ethyl acetate / petroleum ether gradient, at 30 mL / min) to give 950 mg of the title product as a white solid. MS (ES-API pos): 214.2 (M+1) + . Step 2: Methyl 3-acetylbicyclo[1.1.1]pentane-1-carboxylate

[0125] At -78 °C, MeMgBr (3 M, 468.98 μL, 1.5 equiv) was added 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). After the addition was complete, the reaction was slowly warmed to room temperature and stirred at room temperature for 2 h. The reaction was monitored by TLC. After completion of the reaction, the reaction mixture was quenched by adding saturated NH 4 Cl and 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 chromatography ( 4 g Flash silica column, eluent with 0 - 20% ethyl acetate / petroleum ether gradient, at 30 mL / min) to give 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

[0126] In N 2Under an atmosphere at 65 °C, 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 for 2 h. 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 pos): 184.1 (M+1) + 。 Step 4: Methyl 3-(1-aminoethyl)bicyclo[1.1.1]pentane-1-carboxylate

[0127] A mixture of methyl 3-[(Z)-N-hydroxy-C-methyl-carbamimidoyl]bicyclo[1.1.1]pentane-1-carboxylate (70 mg, 382.09 μmol, 1 equiv) and Raney nickel (51.07 mg, 596.06 μmol, 1.56 equiv) in MeOH (3 mL) was degassed and purged with H 2 three times, and then the mixture was stirred at 25 °C under an H 2 atmosphere for 15 h. The reaction was monitored by LCMS. After 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 pos): 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

[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) was added 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). The mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS. After completion of the reaction, 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 pos): 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

[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 H 2 O (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 and 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 then 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% - 72%, 10 min) to give 61.2 mg of the title product as a pale white solid. 1 H NMR(400MHz,CD 3OD) 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 Route 9 Step 1: 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

[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) was added HATU (134.82 mg, 354.56 μmol, 1.2 equiv) and DIPEA (114.56 mg, 886.41 μmol, 154.40 μL, 3 equiv). The mixture was stirred at room temperature for 1 h. The reaction was monitored by LCMS and, after completion, quenched with water and extracted with ethyl acetate (3 × 10 mL). The combined organic layers were concentrated under reduced pressure to afford 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

[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) was added LiOH (68.35 mg, 2.85 mmol, 10 equiv). The mixture was stirred at 50 °C for 2 h. The reaction was monitored by LCMS. After 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. The residue was purified by reverse-phase HPLC (column: Boston Green ODS 150×30 mm X 5 μm; mobile phase: [water (HCl)-ACN]; B%: 50% - 70%, 10 min) to give 47.7 mg of the title product as a pale white solid. 1 H NMR(400MHz,CD 3 OD)δ8.11(br d,J=8.0Hz,1H),7.54(d,J=8.0Hz,2H),7.08(d,J=8.0Hz,2H),6.99(d,J=3.2Hz,1H),6.38(d,J=3.2Hz,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=6.8Hz,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 Route 10 Step 1: 2-Iodo-4-(4-(trifluoromethyl)benzyl)-4H-thieno[3,2-b]pyrrole-3-carboxylic acid

[0132] At 80 °C, a mixture of 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), cobalt dibromide (12.16 mg, 55.59 μmol, 0.1 equiv), 4,7-diphenyl-1,10-phenanthroline (18.48 mg, 55.59 μmol, 0.1 equiv), 167.98 μL, 1.5 equiv) and indigo (255.33 mg, 2.22 mmol, 34.98 μL, 4 equiv) in THF (5 mL) was stirred for 20 h, then the reaction was cooled to room temperature, molecular iodine (211.65 mg, 833.91 μmol) was added, and the reaction was stirred overnight at room temperature. The reaction was monitored by LCMS. After 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%: 62%-92%, 12 min) to give 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 light pink solid. MS(ES-API pos): 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

[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. After completion of the reaction, the reaction was quenched by addition of water, then extracted 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%-83%, 12 min) to give 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 pos): 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

[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) was added CuI (14.64 mg, 76.87 μmol, 3 equiv), methyl 2,2-difluoro-2-fluorosulfonylacetate (14.77 mg, 76.87 μmol, 9.78 μL, 3 equiv). The mixture was then stirred at 100 °C for 16 h. The reaction was monitored by LCMS. After completion of the reaction, 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%-95%, 12 min) to give 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, 55.11% yield) as a white solid. MS(ES-API pos): 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

[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 H 2To the solution in O (0.5 mL), LiOH (4.65 mg, 194.17 μmol, 10 eq) was added. The mixture was stirred at 55 °C for 1 h. The reaction was monitored by LCMS. After completion of the reaction, 1 M HCl was added until pH = 3 - 4 to terminate the reaction, 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% - 76%, 10 min) to give 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, CD 3 OD) 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 pos): 553.2 (M + 1) + 。

[0136] The above examples and descriptions are not intended to limit the scope of the present invention. Any combination of embodiments of the present invention and any obvious extensions or analogs are within the scope of the present invention. In addition, the present invention is intended to cover any arrangement aimed at achieving the same purpose, and all such variations and modifications are within the scope of the appended claims.

Claims

1. A compound of formula I Wherein: R 1 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 attached form a 3- to 6-membered carbocyclic ring, which carbocyclic ring is optionally substituted by one to three R a groups and optionally contains one or two ring-forming heteroatoms, each ring-forming heteroatom independently being S, O or NR b , where each R b 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 is absent, =CH-, -CR 1 R 2 -, or -C(O)-; Cy 1 is C 1-6 alkylene, C 1-6 alkenylene, C 1-6 alkynylene, cycloalkylene, cycloalkenylene, arylene, heteroarylene, heterocycloalkylene, or bridged bicyclo cycloalkylene or cycloalkenylene, and C 1-6 alkylene, C 1-6 alkenylene, C 1-6 each of alkynylene, cycloalkylene, cycloalkenylene, arylene, heteroarylene, heterocycloalkylene, or bridged bicyclo cycloalkylene is optionally substituted; Cy 2 is a cycloalkyl, aryl, heteroaryl or heterocyclic group, and is optionally substituted with one to three substituents, each substituent being independently selected from halogen, alkyl or haloalkyl; and Each R a group is independently halogen, alkyl, haloalkyl, hydroxyalkyl or alkoxy; and when R a is alkyl, Cy 1 is a bridged bicyclic subcycloalkyl; Or a pharmaceutically acceptable salt thereof.

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

3. The compound according to claim 1, wherein Cy 2 is an aryl group and is optionally substituted by a haloalkyl group.

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

5. The compound according to claim 1, wherein Cy 1 is an arylene or a bridged bicyclic subcycloalkyl group.

6. The compound according to claim 1, wherein R 1 and R 2 are each independently hydrogen or C 1-6 alkyl; 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.

7. The compound according to claim 1, wherein R a is -F, -Cl, -CF 3 , hydroxyalkyl, alkoxy or -CH 3 ; and when R a is -CH 3 , Cy 1 is C 5 -C 10 bridged bicyclic subcycloalkyl.

8. The compound according to claim 1, wherein the compound is of formula II where Cy 1 is an arylene or a bridged bicyclic cycloalkylene; and R a is halogen, alkyl, haloalkyl, hydroxyalkyl or alkoxy; when R a is alkyl, Cy 1 is a bridged bicyclic cycloalkylene.

9. The compound according to claim 8, wherein Cy 1 is phenylene or C 5 -C 10 bridged bicyclic subcycloalkyl.

10. The compound according to claim 9, wherein said C 5 -C 10 bridged bicyclic subcycloalkyl is 11. The compound according to claim 1, wherein the compound is 12. A pharmaceutical composition comprising the compound according to any one of claims 1 to 11 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient.

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

14. The pharmaceutical composition according to claim 12 or 13, wherein the composition is used in combination with a radiotherapy agent.

15. A method of treating a subject suffering from a disorder mediated by the action of PGE2 on the EP4 receptor, comprising administering to a subject in need thereof an effective amount of the compound according to any one of claims 1 to 11 or the pharmaceutical composition according to any one of claims 12 to 14.

16. The method according to claim 15, wherein the disorder is 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, allergy, inflammatory bowel disease, interstitial cystitis, mast cell activation syndrome, macrocythemia, 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, gastric cancer, colorectal cancer, liver cancer, pancreatic cancer, skin cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, bone cancer, lymphoma, or blood cancer.

19. Use of the compound according to any one of claims 1 to 11 in the manufacture of a medicament for treating a subject suffering from a disorder mediated by the action of PGE2 on the EP4 receptor.

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

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