CN111936138A - Heterobicyclic Compounds as EP4 Receptor Antagonists - Google Patents

Abstract

The present invention relates to novel EP4 receptor antagonist compounds represented by formula I or a pharmaceutically acceptable salt thereof, which are useful for treating cancer or inflammatory diseases.

Description

Heterobicyclic Compounds as EP4 Receptor Antagonists

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to U.S. Application No. 62/709,882, filed February 5, 2018, the entire contents of which are incorporated herein.

Background technique

The present invention relates to heterocyclic amide derivatives or pharmaceutically acceptable salts thereof, pharmaceutically acceptable prodrugs, pharmaceutical compositions containing them and their medical uses. The compounds of the present invention have activity as prostaglandin E2 receptor antagonists, and they are useful in the treatment or alleviation of pain and inflammation and other diseases associated with inflammation, such as arthritis, in the treatment or prevention of pain, inflammatory diseases and the like disease or medical condition.

Prostaglandins are mediators that trigger pain, fever, and other symptoms related to inflammation. In particular, prostaglandin E2 (PGE2) is the major eicosanoid detected under inflammatory conditions. In addition, PGE2 is also considered to be an important component in the immunosuppressive environment produced by many solid tumors (Whiteside, Expert Opinion in Biotherapy, 2010. 10, 1019-1035), and inhibition of EP4 signaling by antagonists has been shown to reduce tumors Tumor growth in animal models (Terada et al., Cancer Research, 2010, 70, 1606-1615) and tumor metastasis (Yang et al., Cancer Research, 2006, 66, 9665-9672). PGE2 has also been shown to promote the expansion and metastasis of colorectal cancer stem cells in mice (Wang et al., Gastroenterology, 2015, 1-12). Antitumor activity EP4 antagonists have also been shown to ameliorate or synergize the antitumor effects of antibodies against checkpoint proteins, such as anti-CTLA4, anti-PD-L1 or anti-PD-1 (Bao et al., Immunotherapy of Cancer 2015, 3 (Suppl). 2): P35).

The present invention relates to a series of novel EP4 antagonists and methods for the treatment of prostaglandin E2 mediated diseases, particularly for pain, inflammation and cancer immunotherapy, and certain pharmaceutical compositions thereof.

SUMMARY OF THE INVENTION

The present invention relates to a series of novel heterobicyclic derivatives as EP4 receptor antagonists useful in the treatment of EP4 mediated diseases or conditions such as acute and chronic pain, osteoarthritis, rheumatoid arthritis and cancer. Also included are pharmaceutical compositions and methods of use.

On the one hand, the present invention provides the compound shown in formula I or its pharmaceutically acceptable salt or its prodrug,

in:

A, B and C' are each independently selected from N, CH and C(R);

-DEF-Yes-C( ^Rc ) ₂ -N=C-,-N( ^Rc )-C( ^Rc )=C-,-SN=C-,-ON=C-,-N( ^Rc )-N=C-,-C( ^Rc ) ₂ -C( ^Rc ) ₂ -N-,-C( ^Rc )=C( ^Rc )-N-,-C( ^Rc )=NN- , -N=NN-, -OC(R ^c )=C-, or -SC(R ^c )=C-;

R ^1a and R ^2a are each independently H, C _1-6 alkyl, C _1-6 cycloalkyl, C _1-6 fluorocycloalkyl or C _1-6 fluoroalkyl; or R ¹ and R ² are combined with The carbon atoms to which they are all attached together form a three- to six-membered carbocyclic ring, which is ^optionally substituted with R, or form a three- to six-membered ring containing one or two heteroatoms each independently S , S(O) ₂ , S(O)(NH), O, or NR ^e , where R ^e is H, C _1-6 alkyl, C _1-6 cycloalkyl, C _1-6 fluorocycloalkyl , C _1-6 fluoroalkyl, aryl, heteroaryl, C(O)C _1-6 alkyl, C(O) aryl, S(O) ₂ alkyl, or S(O) ₂ aryl ;

each R ^c is independently H, halogen, C _1-4 alkyl, C _1-4 fluoroalkyl, C _1-4 alkoxy, C _1-4 fluoroalkoxy, or acetyl;

each R is independently H, C _1-4 alkyl, C _1-4 fluoroalkyl, C _1-4 alkoxy, C _1-4 fluoroalkoxy, or acetyl;

Y is _-CH2- , -C(O)-, -C(S)-, or -S(O) _2- ;

X is CH ₂ , O, or S;

W is NH or O;

Ar ¹ and Ar ² are each independently C _3-6 cycloalkyl, aryl, heteroaryl or heterocyclyl, or a condensed C _3-6 cycloalkyl, aryl, heteroaryl or heterocyclyl group analogs, and Ar and Ar are each optionally substituted with ^{1-3 R} groups ^;

each R ^b is independently H, halogen, C _1-6 alkyl, C _1-6 alkoxy or C _1-6 fluoroalkyl;

并且 ^Rf is _-CO2H , _-CO2M , or

and

M is a pharmaceutically acceptable salt or ester prodrug group.

In some embodiments, the compound of Formula I is of Formula Ia

in:

A, B and C' are each independently N, CH or C(R);

-DEF-Yes-C( ^Rc ) ₂ -N=C-,-N( ^Rc )-C( ^Rc )=C-,-SN=C-,-ON=C-,-N( ^Rc )-N=C-, -C(R ^c ) ₂ -C(R ^c ) ₂ -N-, -C(R ^c )=C(R ^c )-N-, -C(R ^c )=NN- , -N=NN-, -OC(R ^c )=C-, or -SC(R ^c )=C-;

each Rb is independently H, halogen, C _1-6 alkyl, C _1-6 alkoxy or C _1-6 fluoroalkyl;

R ^1a and R ^2a are each independently H, C _1-6 alkyl, C _1-6 cycloalkyl, C _1-6 fluorocycloalkyl, or C _1-6 fluoroalkyl, or

R ¹ and R ² are each independently H, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 cycloalkyl, C _1-6 fluorocycloalkyl, C _1-6 fluoro Alkyl or _SF5 ^; or R1 and R2 together with the carbon atoms to which they are ^both attached form a three- to six-membered carbon, which may be optionally substituted with ^Rc , or a three-membered ring containing one or two heteroatoms to a six-membered ring, the heteroatoms are each independently S, S(O) ₂ , S(O)(NH), O, or NR ^e , wherein R ^e is H, C _1-6 alkyl, C _{1 -6} cycloalkyl, C _1-6 fluorocycloalkyl, C _1-6 fluoroalkyl, aryl, heteroaryl, C(O)C _1-6 alkyl, C(O) aryl, S( O) ₂ alkyl, or S(O) ₂ aryl;

each Rc is independently H, C _1-4 alkyl, C _1-4 fluoroalkyl, C _1-4 alkoxy, C _1-4 fluoroalkoxy, or acetyl;

each R is independently H, halogen, C _1-4 alkyl, C _1-4 fluoroalkyl, C _1-4 alkoxy, C _1-4 fluoroalkoxy, or acetyl;

X is CH ₂ , O, or S;

each Rb is independently H, halogen, C _1-6 alkyl, C _1-6 alkoxy, or C _1-6 fluoroalkyl;

n is 0, 1, 2, or 3.

In some other embodiments, the compound is of formula Ib:

in:

-DEF- is -C( ^Rc )2-N=C-,-N( ^Rc )-C( ^Rc )=C-,-SN=C-,-ON=C-,-N( ^Rc )-N=C-,-C( ^Rc )2-C( ^Rc )2-N-,-C( ^Rc )=C( ^Rc )-N-,-C( ^Rc )=NN- , -N=NN-, -OC(R ^c )=C-, -SC(R ^c )=C-;

R ¹ and R ² are each independently H, halogen, C _1-6 alkyl, C _1-6 alkoxy, C _1-6 cycloalkyl, C _1-6 fluorocycloalkyl, C _1-6 fluoro Alkyl, or SF ₅ ;

each R ^b is independently H, halogen, C _1-6 alkyl, C _1-6 alkoxy, or C _1-6 fluoroalkyl;

R ^1a and R ^2a are each independently H, C _1-6 alkyl, C _1-6 cycloalkyl, C _1-6 fluorocycloalkyl, or C _1-6 fluoroalkyl; or R ¹ and R ² Together with the carbon atoms to which they are attached, they form a three- to six-membered carbocyclic ring, which may be ^optionally substituted by R, or a three- to six-membered ring containing one or two heteroatoms, which The atoms are each independently S, O or NR ^e , wherein R ^e is H or C _1-6 alkyl;

each R ^c is independently H, C _1-4 alkyl, C _1-4 fluoroalkyl, C _1-4 alkoxy, C _1-4 fluoroalkoxy, or acetyl;

each R is independently H, halogen, C _1-4 alkyl, C _1-4 fluoroalkyl, C _1-4 alkoxy, C _1-4 fluoroalkoxy, or acetyl;

Each R ^b is independently H, halogen, C _1-6 alkyl, C _1-6 alkoxy, or C _1-6 fluoroalkyl; and

n is 0, 1, 2, or 3.

选自以下 杂环部分：In other other embodiments, the fused ring moiety in the compound of formula I

Selected from the following heterocyclic moieties:

Examples of compounds of the present invention include

4-(1-(7-(4-(trifluoromethyl)phenoxy)-1H-indole-1-carboxamido)cyclopropyl)benzoic acid;

4-(1-(7-(3-(trifluoromethyl)phenoxy)-1H-indole-1-carboxamido)cyclopropyl)benzoic acid;

4-(2-(7-(3-(trifluoromethyl)phenoxy)-1H-indole-1-carboxamido)prop-2-yl)benzoic acid;

(±) 4-(1-(7-(3-(trifluoromethyl)phenoxy)-1H-indole-1-carboxamido)ethyl)benzoic acid;

(±) 4-(1-(7-(4-(trifluoromethyl)phenoxy)-1H-indole-1-carboxamido)ethyl)benzoic acid;

(±) 4-(1-(4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)ethyl)benzoic acid;

4-(1-(4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)cyclopropyl)benzoic acid;

4-(1-(4-(3-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)cyclopropyl)benzoic acid;

4-(1-(4-(3-(trifluoromethyl)phenoxy)benzofuran-3-carboxamido)cyclopropyl)benzoic acid;

4-(1-(4-(4-(trifluoromethyl)phenoxy)benzofuran-3-carboxamido)cyclopropyl)benzoic acid;

(±) 4-(1-(4-(4-(trifluoromethyl)phenoxy)benzofuran-3-carboxamido)ethyl)benzoic acid;

(±) 4-(1-(4-(4-(trifluoromethyl)phenoxy)benzo[b]thiophene-3-carboxamido)ethyl)benzoic acid;

(±) 4-(1-(4-(3-(trifluoromethyl)phenoxy)benzo[b]thiophene-3-carboxamido)ethyl)benzoic acid;

4-(1-(4-(3-(trifluoromethyl)phenoxy)benzo[b]thiophene-3-carboxamido)cyclopropyl)benzoic acid;

4-(1-(4-(4-(trifluoromethyl)phenoxy)benzo[b]thiophene-3-carboxamido)cyclopropyl)benzoic acid;

4-(1-(4-(4-(trifluoromethyl)phenoxy)benzo[d]isothiazole-3-carboxamido)cyclopropyl)benzoic acid;

4-(1-(4-(3-(trifluoromethyl)phenoxy)benzo[d]isothiazole-3-carboxamido)cyclopropyl)benzoic acid;

(±) 4-(1-(4-(3-(trifluoromethyl)phenoxy)benzo[d]isothiazole-3-carboxamido)ethyl)benzoic acid;

(±) 4-(1-(4-(4-(trifluoromethyl)phenoxy)benzo[d]isothiazole-3-carboxamido)ethyl)benzoic acid;

(±) 4-(1-(4-(4-(trifluoromethyl)phenoxy)benzo[d]isoxazole-3-carboxamido)ethyl)benzoic acid;

(±) 4-(1-(4-(3-(trifluoromethyl)phenoxy)benzo[d]isoxazole-3-carboxamido)ethyl)benzoic acid;

4-(1-(4-(3-(trifluoromethyl)phenoxy)benzo[d]isoxazole-3-carboxamido)cyclopropyl)benzoic acid;

4-(1-(4-(4-(trifluoromethyl)phenoxy)benzo[d]isoxazole-3-carboxamido)cyclopropyl)benzoic acid;

4-(1-(7-(4-(trifluoromethyl)phenoxy)indoline-1-carboxamido)cyclopropyl)benzoic acid; and

4-(1-(7-(3-(Trifluoromethyl)phenoxy)indoline-1-carboxamido)cyclopropyl)benzoic acid.

As mentioned above, the present invention also includes salts or prodrugs of compounds of formula I. Prodrugs can be selected from esters or amides of compounds of formula I.

In another aspect, the present invention provides pharmaceutical compositions, each comprising a compound as described above and a pharmaceutically or physiologically acceptable carrier.

The present invention also relates to a method for treating a subject with a disease mediated by the action of PGE2 on the EP4 receptor. The method includes administering to a subject in need thereof an effective amount of a compound or pharmaceutical composition described above.

In some embodiments, the disorder is an inflammatory disease or cancer. Examples of inflammatory diseases include arthritis, acne vulgaris, asthma, autoimmune diseases, autoinflammatory diseases, celiac disease, chronic prostatitis, colitis, diverticulitis, glomerulonephritis, hidradenitis seborrheic, allergic reactions , inflammatory bowel disease, interstitial cystitis, mast cell activation syndrome, mastocytosis, otitis media, pelvic inflammatory disease, reperfusion injury, rheumatic fever, rheumatoid arthritis, rhinitis, sarcoidosis and vasculitis.

In some other embodiments, the disorder is cancer, and the method comprises administering to a subject in need thereof an effective amount of a compound or composition described above, optionally in combination with a second therapeutic agent, the second The therapeutic agent is an antibody, kinase inhibitor, IDO inhibitor, TDO inhibitor, STING activator, HDAC inhibitor or chemotherapeutic agent. Examples of suitable antibodies include those against CTCLA4, PDL1 and PD1.

Detailed ways

definition

Abbreviations used herein have their conventional meanings in the fields of chemistry and biology. .

As used herein, the term "alkyl" by itself or as part of another substituent, unless otherwise specified, refers to a straight chain (ie, unbranched) or branched chain, or a cyclic hydrocarbon group, or a combination thereof, which may be fully Saturated, monounsaturated or polyunsaturated, can include divalent and polyvalent groups having the specified number of carbon atoms (ie, C1-C10 represents 1 to 10 carbon atoms). Examples of saturated hydrocarbon groups include, but are not limited to, such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclohexyl Propylmethyl, a homologous group. And isomers such as n-pentyl, n-hexyl, n-heptyl, n-octyl and the like. Unsaturated alkyl groups are alkyl groups having one or more double or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, crotyl, 2-prenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1 , 4-pentadienyl), ethynyl, 1- and 3-propynyl, 3-butynyl and higher homologues and isomers. Alkyl groups restricted to hydrocarbyl groups are referred to as "pure alkyl groups".

As used herein, the term "fluoroalkyl" refers to an alkyl group as defined above wherein one or more hydrogen atoms have been replaced with fluorine atoms.

As used herein, the term "alkylene" by itself or as part of another substituent refers to a _divalent group derived from an alkyl group such as, but _{not limited} to, _{-CH2CH2CH2CH2-} , -CH ₂ CH=CHCH ₂ -, -CH ₂ C≡CCH ₂ -, -CH ₂ CH ₂ CH(CH ₂ CH ₂ CH ₃ )CH ₂ -. Typically, alkyl groups (or alkylene groups) have 1 to 24 carbon atoms, and those groups having 10 or fewer carbon atoms are preferred in the present invention. "Lower alkyl" or "lower alkylene" is a shorter chain alkyl or alkylene group, usually having eight or fewer carbon atoms.

As used herein, the term "alkynyl" refers to a carbon chain containing at least one carbon-carbon triple bond, which may be straight or branched or a combination thereof. Examples of alkynyl groups include ethynyl, propargyl, 3-methyl-1-pentynyl, 2-heptynyl and the like.

As used herein, the term "cycloalkyl" refers to a monocyclic or bicyclic saturated carbocycle, each carbocycle having 3 to 10 carbon atoms. A "fused analog" of a cycloalkyl group refers to a single ring fused to an aryl or heteroaryl group where the point of attachment is on the non-aromatic moiety. Examples of cycloalkyl and fused analogs thereof include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, tetrahydronaphthyl, decalinyl, indanyl, and the like.

As used herein, the term "alkoxy" refers to a straight or branched chain alkoxy group having the indicated number of carbon atoms. C _1-6 alkoxy, for example, includes methoxy, ethoxy, propoxy, isopropoxy and the like. .

The term "heteroalkyl" as used herein, by itself or in combination with another term, means, unless otherwise specified, consisting of at least one carbon atom and at least one heteroatom selected from O, N, P, Si, and S A stable linear or branched or cyclic hydrocarbon group or a combination thereof, wherein the nitrogen, phosphorus and sulfur atoms are optionally oxidized, and the nitrogen heteroatoms are optionally quaternized. The heteroatoms O, N, P and S and Si can be located at any internal position of the heteroalkyl group or the position where the alkyl group is attached to the rest of the molecule. Examples include, but are not limited to -CH ₂ -CH ₂ -O-CH ₃ , -CH ₂ -CH ₂ -NH-CH ₃ , -CH ₂ -CH ₂ -N(CH ₃ )-CH ₃ , -CH ₂ -S -CH ₂ -CH ₃ , -CH ₂ -CH ₂ , -S(O)-CH ₃ , -CH ₂ -CH ₂ -S(O) ₂ -CH ₃ , -CH=CH-O-CH ₃ , - Si( _CH3 ) ₃ , _-CH2 -CH=N- _OCH3 , -CH=CH-N( _CH3 ) _-CH3 , -O- _CH3 , -O- _CH2 - _CH3 , and -CN . Up to two or three heteroatoms may be consecutive, eg _-CH2 -NH- _OCH3 and _-CH2 -O-Si( _CH3 ) ₃ . Similarly, the term "heteroalkylene" by itself or as part of another substituent refers to a divalent group derived from a heteroalkyl group such as, but not limited to, _-CH2 - _CH2 -S- _CH2 -CH2- and _{-CH2 -} S- _CH2 - _{CH2 -} NH-CH2-. For heteroalkylene groups, heteroatoms may also occupy one or both of the chain termini (eg, alkyleneoxo, alkylenedioxo, alkyleneamino, alkylenediamino, and the like). Furthermore, for alkylene and heteroalkylene linking groups, the chemical formula of the linking group is written in no way implying the orientation of the linking group. For example, the formula -C(O)OR'- represents both -C(O)OR'- and -R'OC(O)-. As mentioned above, heteroalkyl groups as used herein include those groups attached to the remainder of the molecule through a heteroatom, eg, -C(O)R', -C(O)NR', -NR'R", - OR', -SR', and/or _-SO2R '. Where a "heteroalkyl" is recited, followed by a specific heteroalkyl, eg, -NR'R" or the like, the term heteroalkyl is to be understood and -NR'R" are not redundant or mutually exclusive. Rather, specific heteroalkyl groups are listed for added clarity. The term "heteroalkyl" should not be construed herein to exclude specific heteroalkyl groups such as -NR'R" or the like.

As used herein, the term "cycloalkoxy" refers to a cycloalkyl group, as defined above, bonded to an oxygen atom, eg, cyclopropoxy.

As used herein, the term "fluoroalkoxy" refers to an alkoxy group as defined above wherein one or more hydrogen atoms have been replaced with fluorine atoms.

As used herein, the term "aryl" refers to a monocyclic or bicyclic aromatic ring containing only carbon atoms. A "fused analog" of an aryl group refers to an aryl group fused to a monocyclic cycloalkyl or monocyclic heterocyclyl group, wherein the point of attachment is on the aromatic moiety. Examples of aryl groups and their fused analogs include phenyl, naphthyl, indanyl, indenyl, tetrahydronaphthyl, 2,3-dihydrobenzofuranyl, dihydrobenzopyranyl, 1, 4-Benzodioxanyl and the like.

As used herein, the term "heteroaryl" refers to a monocyclic or bicyclic aromatic ring containing at least one heteroatom selected from N, O and S, each ring containing 5 to 6 atoms. A "fused analog" of a heteroaryl group refers to a heteroaryl group fused to a monocyclic cycloalkyl or monocyclic heterocyclyl, wherein the point of attachment is on the aromatic moiety. Examples of heteroaryl groups include pyrrolyl, isoxazolyl, isothiazolyl, pyrazolyl, pyridyl, oxazolyl, oxadiazolyl, thiadiazolyl, thiazolyl, imidazolyl, triazolyl, tetrazolyl azolyl, furyl, triazinyl, thienyl, pyrimidinyl, pyridazinyl, pyrazinyl, benzoxazolyl, benzothiazolyl, benzimidazolyl, benzofuranyl, benzothienyl, Furan(2,3-b)pyridyl, quinolinyl, indolyl, isoquinolinyl, and the like.

The aryl and heteroaryl groups referred to in the definitions of Ar ¹ and Ar ² are unsubstituted or substituted with at least one substituent selected from substituents.

Substituents are selected from halogen atoms, alkyl groups having 1-4 carbon atoms, alkoxy groups having 1-4 carbon atoms, haloalkyl groups having 1-4 carbon atoms, haloalkanes having 1-4 carbon atoms Oxy, cyano, alkynyl with 2 to 6 carbon atoms, alkanoyl with 1 to 5 carbon atoms, cycloalkyl with 3 to 7 ring atoms, heteroaryl, aryl, with 7 to 7 Aralkoxy of 10 carbon atoms, arylcarbonyl, two adjacent x groups optionally joined together to form an alkylene or alkenylene chain with 3 or 4 carbon atoms, aminocarbonyl, Alkenyl having 2 to 5 carbon atoms, alkylthio having 1 to 4 carbon atoms, aminosulfinyl, aminosulfonyl, hydroxy, hydroxyalkyl having 1 to 4 carbon atoms, nitro, amino , carboxyl, alkoxycarbonyl having 2 to 5 carbon atoms, alkoxyalkyl having 1-4 carbon atoms, alkylsulfonyl having 1-4 carbon atoms, alkoxycarbonyl having 1-4 carbon atoms alkanoylamino, alkanoyl(alkyl)amino having 1-6 carbon atoms, alkanoylaminoalkyl having 1-6 carbon atoms in the alkanoyl and alkyl moieties, in the alkanoyl and each alkyl alkanoyl(alkyl)aminoalkyl having 1 to 6 carbon atoms in the moiety, alkylsulfonylamino having 1 to 4 carbon atoms, mono or dialkylaminocarbonyl having 1 to 6 carbon atoms, Mono- or dialkylaminosulfinyl having 1-6 carbon atoms, mono- or dialkylaminosulfonyl having 1-6 carbon atoms, aminoalkyl having 1-4 carbon atoms, having 1 to Mono- or di-alkylamino groups of 6 carbon atoms, mono- or di-alkylaminoalkyl groups of 1 to 6 carbon atoms in each alkyl moiety, aralkyl groups of 7 to 10 carbon atoms, in Heteroarylalkyl having 1 to 4 carbon atoms in the alkyl group, heteroarylalkoxy having 1 to 4 carbon atoms in the alkoxy moiety, and alkylsulfonic acid having 1 to 4 carbon atoms Acylamino.

The term "heterocyclyl" as used herein refers to a monocyclic or bicyclic saturated ring containing at least one heteroatom selected from N, S and O, each of said rings having from 3 to 10 atoms, wherein the point of attachment may be is carbon or nitrogen. A "fused analog" of a heterocyclyl group refers to a monocyclic heterocycle fused to an aryl or heteroaryl group where the point of attachment is on the non-aromatic moiety. Examples of "heterocyclyl" and fused analogs thereof include pyrrolidinyl, piperidinyl, piperazinyl, imidazolidinyl, 2,3-dihydrofuran(2,3-b)pyridyl, benzoxanyl Azinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, indoline and the like. The term also includes partially unsaturated monocyclic rings that are not aromatic, such as 2- or N-substituted-(1H,3H)-pyrimidine-2,4-dione (N-substituted uracil) linked via nitrogen or N-substituted uracil 4-pyridone.

The term "halo" or "halogen" as used herein, by itself or as part of another substituent, refers to a fluorine, chlorine, bromine or iodine atom, unless otherwise indicated. Additionally, terms such as "haloalkyl" are meant to include monohaloalkyl and polyhaloalkyl. For example, the term "halo(C1-C4)alkyl" is meant to include, but not be limited to, trifluoromethyl, 2,2,2-trifluoroethyl, 4-chlorobutyl, 3-bromopropyl, and the like thing.

As used herein, the term "prodrug" refers to an agent that is converted to the parent drug in vivo. Prodrugs are often useful because in some cases they may be easier to administer than the parent drug. For example, they may be bioavailable by oral administration, while the parent is not. The prodrugs can also have improved solubility in pharmaceutical compositions compared to the parent drug. Without limitation, an example of a prodrug is any one of the compounds of formula I, which is administered as an ester ("prodrug") to facilitate delivery across cell membranes, where water solubility is not conducive to migration, but once inside the cell In this context, water solubility is favorable, and the ester is then metabolically hydrolyzed to the active carboxylic acid. Another example of a prodrug might be a short peptide (polyamino acid) bound to an acidic group, where the peptide is metabolized to reveal the active moiety.

Optical isomers - diastereomers - geometric isomers - tautomers

The compounds of formula I contain one or more asymmetric centers and thus can exist as racemates and racemic mixtures, single enantiomers, diastereomeric mixtures and individual diastereomers. The present invention is intended to understand all such isomeric forms of the compounds of formula I.

Some of the compounds described herein contain olefinic double bonds, and unless otherwise specified, are meant to include both E and Z geometric isomers.

Certain compounds described herein may exist at different points of hydrogen attachment, termed tautomers. Examples of this would be ketones and their enol forms, known as keto-enol tautomers. Compounds of Formula I to Formula Ig encompass individual tautomers and mixtures thereof.

Compounds of formula I can be separated into diastereomeric pairs of enantiomers, for example, by fractional crystallization from suitable solvents such as MeOH or EtOAc or mixtures thereof. The enantiomeric pairs obtained can be separated into the individual stereoisomers by conventional means, for example by using optically active amines as resolving agents or on chiral HPLC columns.

Alternatively, any enantiomer of a compound of general formula I can be obtained by stereospecific synthesis using optically pure starting materials or reagents of known configuration.

salt

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. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, iron, ferrous, lithium, magnesium, manganese, manganese, potassium, sodium, zinc and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases including salts of primary, secondary and tertiary amines including naturally occurring substituted amines, cyclic amines and substituted amines of basic ion exchange resins such as arginine, betaine , caffeine, choline, N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylamine Phytoline, N-ethylpiperidine, glucosamine (glucamine), glucosamine (glucosamine), histidine, hydrazine diamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, Piperidine, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine, etc.

When the compounds of the present invention are basic, salts can be prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include acetic acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, citric acid, ethanesulfonic acid, fumaric acid, gluconic acid, glutamic acid, hydrobromic acid, hydrochloric acid, plasma, lactic acid, maleic acid, apple acid, mandelic acid, methanesulfonic acid, viscose, nitric acid, pamoic acid, pantothenic acid, sulfuric acid, phosphoric acid, succinic acid, tartaric acid, p-toluenesulfonic acid, and the like. Particularly preferred are citric acid, hydrobromic acid, hydrochloric acid, maleic acid, phosphoric acid, sulfuric acid and tartaric acid.

It should be understood that, as used herein, references to compounds of formula I are also meant to include pharmaceutically acceptable salts.

practical

The compounds of the present invention are antagonists of the EP4 receptor and are therefore useful in the treatment of prostaglandin E2 mediated diseases or disorders.

Merck)。The present invention also includes methods of treating cancer, using an effective amount of a compound of the present invention or using an effective amount of a compound of the present invention in combination with an effective amount of radiation; antibodies to cytotoxic T-lymphocyte antigen 4 (anti-CTLA4); programmed death ligand 1 Antibody (anti-PDL1); Antibody to programmed cell death protein 1 (anti-PD1); Indoleamine-2,3-dioxygenase (IDO) inhibitor; Tryptophan-2,3-dioxygenase ( TDO) inhibitors; and antimetabolites. These antibodies may be selected from, but are not limited to, MDX-010 (ipilimumab, Bristol-Myers Squibb), CP-675, 206 (tremelimumab, Pfizer), MPDL3280A (Roche), MDX-1106 (nivolumab, Bristol-Myers Squibb), labrolizumab (Merck) and pembrolizumab (

Merck).

In another aspect of the invention, the cancer to be treated is selected from the group consisting of breast cancer, cervical cancer, colorectal cancer, endometrial cancer, glioblastoma, head and neck cancer, kidney cancer, liver cancer, lung cancer, medulloblastoma , ovarian, pancreatic, prostate, skin and urinary tract cancers. In a more specific aspect of the present invention, there is provided a method of treating cancer and/or generating a memory immune response comprising administering a compound of formula I.

As used herein, the term "treating a disease or disorder mediated by prostaglandin E2" refers to the treatment or prevention of any chronic disease or disorder which can be beneficially treated or prevented by selecting an EP4 antagonist. The term includes relief of pain, fever, and inflammation in a variety of ailments, including rheumatic fever, symptoms associated with influenza or other viral infections, the common cold, lower back pain, neck pain, dysmenorrhea, headaches, migraines, toothaches , sprains and strains, rheumatoid arthritis, myositis, neuralgia, synovitis, arthritis (including rheumatoid arthritis), degenerative joint disease (osteoarthritis), gout, ankylosing spondylitis, bursae inflammation, burns, injuries, and pain and inflammation after surgery. In addition, such compounds inhibit neoplastic transformation of cells and metastatic tumor growth and are therefore useful in the treatment and/or prevention of cancer.

As used herein, the term "effective amount" or "therapeutically effective amount" refers to an amount effective to treat a prostaglandin E2-mediated disease or disorder or cancer, as indicated by clinical testing and evaluation, patient observation, and/or the like quantity. An "effective amount" can further refer to an amount that causes a detectable change in biological or chemical activity. Those skilled in the art can detect and/or further quantify detectable changes based on the relevant mechanism or process. Also, an "effective amount" can refer to an amount that maintains a desired physiological state, i.e., reduces or prevents a significant decrease in the condition and/or promotes an improvement in the condition. An "effective amount" can further refer to a therapeutically effective amount.

As used herein, the term "cancer" can include cancers that result from inherited mutations in genes. Examples of such cancers include, but are not limited to, breast cancer, cancers associated with Lee-Fraumeni syndrome, such as childhood sarcoma, leukemia, and brain cancer, cancers associated with Lynch syndrome, such as colon cancer, bile duct cancer, Brain, Endometrial, Kidney, Ovarian, Pancreatic, Small Intestine, Stomach and Ureteral, Lung, Melanoma, Prostate, Retinoblastoma, Thyroid, and Uterine Cancers. In addition, cancer can be the result of acquired mutations, e.g., mutations caused by diet, environment and/or lifestyle, or somatic mutations. Examples of such cancers may include, but are not limited to, adrenal cancer, adrenocortical cancer, 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 cancer), endometrial cancer, epidermal cancer, esophagus cancer, gallbladder cancer, genitourinary tract cancer, head and neck cancer, kidney cancer, liver cancer, lung cancer (non-limiting examples include adenocarcinoma, small cell lung cancer and non-small cell lung cancer), lymphoma (non-limiting examples include B-cell lymphoma, T-cell lymphoma, Hodgkin lymphoma, non-Hodgkin lymphoma), melanoma, malignant Melanoma, malignant carcinoid, malignant pancreatic insulinoma, myeloma, multiple myeloma, ovarian cancer, pancreatic cancer (e.g. exocrine pancreatic cancer), prostate cancer, renal cell carcinoma, skin cancer, in addition to other cancers mentioned above In addition, also includes squamous cell carcinoma, gastric carcinoma, testicular carcinoma, thyroid carcinoma, thyroid follicular carcinoma, Wilms tumor, choriocarcinoma, mycosis, malignant hypercalcemia, cervical hyperplasia, leukemia, acute lymphoid Cellular leukemia, chronic lymphocytic leukemia, hairy cell lymphoma, Burke's lymphoma, acute myeloid leukemia, chronic myeloid leukemia, myelodysplastic syndrome, promyelocytic leukemia, chronic myeloid leukemia, acute myeloid leukemia Cellular leukemia, fibrosarcoma, rhabdomyosarcoma, astrocytoma, neuroblastoma, rhabdomyosarcoma, schwannoma, Kaposi's sarcoma, polycythemia vera, essential thrombocythemia, Hodgkin's disease, non- Hodgkin lymphoma, soft tissue sarcoma, osteosarcoma, primary macroglobulinemia, seminoma, teratoma, osteosarcoma, xenocarcinoma, keratokeratoma, and retinoblastoma.

The compound shown in formula I can also be used in combination with one or more chemotherapeutic agents, such as:

i. aromatase inhibitors,

ii. Antiestrogens, antiandrogens (especially in the case of prostate cancer) or gonadotropin agonists,

iii. a topoisomerase I inhibitor or a topoisomerase II inhibitor,

iv. Microtubule active agents, alkylating agents, antineoplastic antimetabolites or platinum compounds,

v. Compounds that target/reduce protein or lipid kinase activity or protein or lipid phosphatase activity, further anti-angiogenic compounds or compounds that induce cellular differentiation processes,

vi. a bradykinin I receptor or angiotensin II antagonist,

vii. Cyclooxygenase inhibitors, bisphosphonates, rapamycin derivatives such as everolimus, heparanase inhibitors (prevents degradation of heparan sulfate) such as P188, a biological response modifier, Preferably immunoglobulins or interferons, eg, interferons, ubiquitin inhibitors or inhibitors that block anti-apoptotic pathways,

viii. Inhibitors of oncogenic isoforms of Ras, eg. H-Ras, K-Ras or N-Ras, or farnesyltransferase inhibitors such as L-744, 832 or DK8G557,

ix. Telomerase inhibitors, such as telomestatin,

x. protease inhibitors, matrix metalloproteinase inhibitors, methionine aminopeptidase inhibitors, such as benzamide or derivatives thereof, or proteosome inhibitors, such as PS 341,

xi. Histone deacetylase inhibitors, such as Vorinostat, MG0103 or MS275, and

xii. Kinase inhibitors.

The pharmaceutical composition of the present invention comprises the compound shown in formula I or a pharmaceutically acceptable salt thereof as an active ingredient, and can also comprise a pharmaceutically acceptable carrier and optional other therapeutic ingredients. The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases, including inorganic and organic bases. Salts derived from inorganic bases include aluminum, ammonium, calcium, copper, iron, ferrous, lithium, magnesium, manganese, manganese, potassium, sodium, zinc and the like. Particularly preferred are the ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary and tertiary amines, including naturally occurring substituted amines, cyclic amines and substituted amines of basic ion exchange resins such as arginine, betaine, Caffeine, Choline, N,N-Dibenzylethylenediamine, Diethylamine, 2-Diethylaminoethanol, 2-Dimethylaminoethanol, Ethanolamine, Ethylenediamine, N-Ethylmorpholine, N-Ethylpiperidine, Glucosamine, Glucosamine, Histidine, Hydrazine, Isopropylamine, Lysine, Methylglucamine, Morpholine, Piperazine, Piperidine, Polyamine Resin, Polyamine Lucaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, tromethamine and the like.

It should be understood that in the discussion of methods of treatment that follow, references to compounds of formula I are also meant to include pharmaceutically acceptable salts.

EP4 antagonists, antibodies and/or antimetabolites can be administered to a subject by any suitable route, including orally (including by buccal administration, but also by oral gastrogastric tube), intraperitoneally, parenterally, The spray is inhaled through topical (ie, skin and mucosal surfaces, including airway surfaces), transdermally, rectally, nasally (including nasogastric feeding tubes), sublingually, buccally, vaginally, or through an implantable reservoir. The term "parenteral" as used herein includes subcutaneous, intramuscular, intradermal, intravenous, intraarticular, 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.

Pharmaceutical compositions containing the active ingredient may be in a form suitable for oral use such as tablets, troches, troches, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules or syrups or Elixirs. Compositions for oral administration may be prepared according to any method known in the art for the manufacture of pharmaceutical compositions, and these compositions may contain one or more selected from the group consisting of sweetening agents, flavoring agents, coloring agents and preservatives agents to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with non-toxic pharmaceutically acceptable excipients which are suitable for the manufacture of tablets. These excipients may 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; binders, such as starch, gelatin Or gum arabic, and lubricants such as magnesium stearate, stearic acid, or talc. Tablets may be uncoated or coated by known techniques to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period of time. For example, time delay materials such as glyceryl monostearate or glyceryl distearate can be used. They can also be coated by the techniques described in U.S. Patents 4,256,108; 4,166,452 and 4,265,874 to form osmotic therapeutic tablets for controlled release.

Oral preparations can also be presented as hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate or kaolin, or soft gelatin capsules in which the active ingredient is mixed with an aqueous or oily vehicle, such as peanut oil , liquid paraffin or olive oil.

Aqueous suspensions contain the active materials in admixture with excipients suitable for the manufacture of aqueous suspensions. Such excipients are suspending agents such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, xanthan gum and acacia. The dispersing or wetting agent can be a naturally occurring phospholipid, such as lecithin, or the condensation product of an alkylene oxide with a fatty acid, such as polyoxyethylene stearate, or the condensation product of ethylene oxide with a long-chain aliphatic alcohol, For example, heptane ethylene-oxycetyl alcohol, or the condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols, such as polyoxyethylene sorbitan monooleate, or ethylene oxide with derivatives derived from fatty acids and hexitols Condensates of partial esters of anhydrous sugar alcohols, such as polyethylene sorbitan monooleate. The aqueous suspension may also contain one or more preservatives, such as ethyl or n-propyl p-hydroxybenzoate, one or more coloring agents, one or more flavoring agents, and one or more Various sweeteners such as sucrose, saccharin or aspartame.

Oil suspensions can be formulated by suspending the active ingredient in a vegetable oil such as peanut oil, olive oil, sesame oil or coconut oil, or a mineral oil such as liquid paraffin. Oily suspensions may contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Sweetening and flavoring agents such as those described above may be added to provide a palatable oral preparation. These compositions can be preserved by adding antioxidants such as ascorbic acid.

Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water provide the active ingredient in admixture with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Other excipients such as sweetening, flavouring and colouring agents may also be present.

The pharmaceutical compositions of the present invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil such as olive or peanut oil, or a mineral oil such as liquid paraffin or a mixture of these. Suitable emulsifiers may be naturally occurring phospholipids, such as soybeans, lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, such as sorbitan monooleate, and the partial esters with ethylene oxide. The condensation products of , such as polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening and flavoring agents.

Syrups and elixirs can be formulated with sweetening agents such as glycerol, propylene glycol, sorbitol or sucrose. Such formulations may also contain a demulcent, a preservative and flavoring and coloring agents. Pharmaceutical compositions can be in the form of sterile injectable aqueous or oily suspensions. This suspension may be formulated according to the known art using those suitable dispersing or wetting agents and suspending agents which have been mentioned above. The sterile injectable preparation may also be a sterile injectable solution or suspension in a nontoxic parenterally acceptable diluent or solvent, for example as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution and isotonic sodium chloride solution. In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium. For this purpose any bland fixed oil can be employed including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid find use in the preparation of injectables.

The compounds of the invention may also be administered intranasally or by inhalation, usually in dry powder form from a dry powder inhaler (alone, as a mixture, for example, with lactose, or as mixed-component particles, for example, with phospholipids, such as phospholipids acylcholine mix), or from a pressurized container, pump, nebulizer, nebulizer (preferably an I nebulizer that produces a fine mist using electrohydrodynamics), or a nebulizer with or without a suitable propellant In the form of an aerosol, a propellant such as 1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane. For intranasal use, the powder may contain a bioadhesive such as chitosan or cyclodextrin.

Pressurized container, pump, nebulizer, atomizer or nebuliser containing a solution or suspension of a compound of the present invention, said solution or suspension containing, for example, ethanol, an aqueous ethanol solution or for dispersion , suitable substitutes for solubilizing or prolonging the release of active substances. One or more propellants are used as solvent and an optional surfactant such as sorbitan trioleate, oleic acid or oligomeric lactic acid.

The drug product is micronized to a size suitable for delivery by inhalation (usually less than 5 microns) prior to use in dry powder or suspension formulations.

This can be achieved by any suitable pulverization method, such as spiral jet milling, fluidized bed jet milling, supercritical fluid treatment to form nanoparticles, high pressure homogenization or spray drying.

Capsules for inhalers or insufflators (for example, made of gelatin or HPMC), blisters and cartridges can be formulated to contain a powder mixed with a compound of the invention, a suitable powder base such as lactose or starch, and a performance modifier , such as 1-leucine, mannitol or magnesium stearate. Lactose can be either anhydrous or in the form of a monohydrate, the latter being preferred. Other suitable excipients include dextran, glucose, maltose, sorbitol, xylitol, fructose, sucrose and trehalose.

Suitable solution formulations for use in nebulizers that use electrohydrodynamics to generate fine mists may contain from log to 20 mg of a compound of the invention per actuation, and the actuation volume may vary from 11 to 1001. A typical formulation may contain the compound of formula (1), propylene glycol, sterile water, ethanol and sodium chloride. Alternative solvents that can be used in place of propylene glycol include glycerol and polyethylene glycol.

Suitable flavoring agents, such as menthol and levomenthol, or sweetening agents, such as saccharin or sodium saccharin, may be added to those formulations of the invention intended for inhalation/intranasal administration.

Formulations for inhalation/intranasal administration can be formulated as immediate and/or sustained release formulations using, for example, poly(DL-lactic-glycolic acid (PGLA)). Sustained-release formulations include delayed-, sustained-, pulsed-, controlled-, targeted and programmed release.

In the case of dry powder inhalers and aerosols, the dosage unit is determined by a valve that delivers a metered dose. Units according to the present invention are generally arranged to administer a metered dose or "puff" containing 1 to 10 mg of the compound of formula I. The total daily dose is usually in the range of 1 to 10 mg and can be administered in a single dose or, more commonly, in divided doses throughout the day.

The compounds of formula I may also be administered in the form of suppositories for rectal administration. These compositions can be prepared by mixing the drug with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Such materials are cocoa butter and polyethylene glycols.

For topical use, creams, ointments, jellies, solutions or suspensions, etc., containing the compound of formula I are used. (For the purposes of this application, topical application shall include mouthwashes and gargles.)

Dosage levels of from about 0.01 mg to about 140 mg/kg body weight per day can be used to treat the aforementioned conditions, or from about 0.5 mg to about 7 g per patient per day. For example, inflammation can be effectively treated by administering about 0.01 to 50 mg of the compound per kilogram of body weight per day, or about 0.5 mg to about 3.5 g of the compound per patient per day, preferably 2.5 mg to 1 g per patient per day.

The amount of active ingredient that can be combined with a carrier material to produce a single dose will vary depending upon the host treated and the particular mode of administration. For example, a formulation for human oral administration may contain from 0.5 mg to 5 g of the active agent, together with a suitable and convenient amount of carrier material, which may comprise from about 5% to about 95% of the total amount of the composition. Dosage unit forms will generally contain from about 1 mg to about 500 mg of active ingredient, usually 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg or 1000 mg.

Assays to measure biological activity

The compounds of formula I can be tested for their prostaglandin antagonist or agonist activity and selectivity in vitro and in vivo using the following assays. Prostaglandin receptors exhibiting activity are DP, EP1, EP2, EP3, EP4, FP, IP and TP.

Stable expression of prostaglandin receptors in human embryonic kidney (HEK) 293 (ebna) cell line

The prostaglandin receptor cDNA corresponding to the full-length coding sequence was subcloned into the appropriate site in a mammalian expression vector and transfected into HEK 293 (ebna) cells. HEK 293 (ebna) cells expressing a single cDNA were grown under selective conditions, and after 2-3 weeks of growth, single colonies were isolated using the cloning circle method and then expanded into clonal cell lines.

Prostaglandin receptor binding assay

Transfected HEK 293(ebna) cells were maintained in culture, harvested, and membranes prepared by differential centrifugation, and cells were lysed in the presence of protease inhibitors 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) (EPs, FP and TP) or Performed in 10 mM HEPES/KOH (pH 7.4) (DP and IP) containing 1 mM EDTA, 2.5-30 mM divalent cations and appropriate radioligands. Synthetic compounds were added to dimethyl sulfoxide and kept constant at 1% (v/v) in all incubations. Reactions are initiated by the addition of membrane proteins. Nonspecific binding was determined in the presence of 10 [mu]M of the corresponding nonradioactive prostanoids. Incubate for 60-90 min at room temperature or 30°C and terminate by rapid filtration. Specific binding was calculated by subtracting nonspecific binding from total binding. Residual specific binding at each ligand concentration was calculated and expressed as a function of ligand concentration to construct a sigmoid concentration-response curve. Binding affinity of a compound is determined by calculating the equilibrium inhibition constant (Ki) from the equation Ki=InPt/1+Radioligand/Kd, where Kd is the equilibrium dissociation constant for the radioligand:receptor interaction and InPt is the dose-response curve the turning point.

The EP4 receptor binding assay was performed at the MSD Pharma Service in Taiwan, China under the following assay conditions:

Source: Human recombinant Chem-1 cells

Ligand: 1 nM [ ³ H]prostaglandin E ₂ (PGE ₂ )

Vehicle: 1% DMSO

Incubation time/temperature: 2 hours, 25°C

Incubation buffer: 10 mM MES, pH 6.0, 1 mM EDTA, 10 mM MgCl.

Nonspecific ligand: 10 μM prostaglandin E ₂ (PGE ₂ )

KD: _0.69nM

B _max : 4.3pmole/mg protein*

Specific binding: 90%

Quantitative Methods: Radioligand Binding

Significance criteria: >50% maximal inhibition

Table 1. Inhibition of PGE2 Binding by Representative Compounds

Prostaglandin receptor agonist and antagonist assay

Stimulation of intracellular cAMP accumulation in HEK-293(ebna)-hEP4 cells was measured using a whole cell second messenger assay to determine whether receptor ligands were agonists or antagonists. Cells were harvested and resuspended in HBSS containing 25 mM HEPES, pH 7.4. Incubations contained 0.5 mM IBMX (phosphodiesterase inhibitor, available from Biomol). The samples were incubated at 37°C for 10 minutes, the reaction was terminated, and cAMP levels were measured. Ligands were added to dimethyl sulfoxide and kept constant at 1% (v/v; agonist) or 2% (v/v; antagonist) in all incubations. For agonists, second messenger responses were expressed as a function of ligand concentration and _EC50 values and maximal responses were calculated compared to PGE ₂ standards. For antagonists, the ability of the ligand to inhibit the agonist response was determined by performing dose _- response curves in the presence of the PGE2 agonist at a concentration corresponding to its _EC70 . _IC50 values were calculated as the ligand concentration required to inhibit 50% of PGE2 _- induced activity.

Rat paw edema test

This method is the same as that described by Chan et al. (J. Pharmacol. Exp. Ther. 274: 1531-1537, 1995).

Carrageenan induces acute inflammatory hyperalgesia in rats

The method is the same as described by Boyce et al. (Neuropharmacology 33: 1609-1611, 1994).

Adjuvant-induced arthritis in rats

Female Lewis rats (about 146-170 g body weight) were weighed, ear-tagged, and grouped (negative control group without induced arthritis, vehicle control group, and positive control group with a total dose of 1 mg/kg of indomethacin per day). The four groups were administered with a total daily dose of 0.10-3.0 mg/kg of the test compound), so that the body weight of each group was equal. 0.5 mg of Mycobacterium butyricum (adjuvant) in 0.1 mg of light mineral oil was injected into the hind paws of six groups of 10 rats, while the negative control group of 10 rats was not injected with adjuvant. agent. Body weight, contralateral paw volume (determined by mercury displacement plethysmography), and lateral radiographs (obtained under ketamine and xylazine anesthesia) were determined 1 day before adjuvant injection and 21 days after injection, and were injected Primary paw volume was determined before (day -1) and on days 4 and 21 after adjuvant injection. Rats were anesthetized with a combined intramuscular injection of 0.03-0.1 mL of ketamine (87 mg/kg) and xylazine (13 mg/kg) for radiography and adjuvant injections. Radiographs were made on days 0 and 21 with both hind paws using Faxitron (45kVp, 30 seconds) and Kodak X-OMAT TL film and developed in an automatic processor. Radiographs were assessed for soft and hard tissue changes by an investigator reluctant to perform the experimental treatment. The following radiographic changes are numerically graded according to severity: increased soft tissue volume (0-4), reduced or enlarged joint space (0-5), subchondral erosion (0-3), periosteal reaction (0-4), bone Lysis (0-4), subluxation (0-3), joint degenerative changes (0-3). A numerical rating of the severity of each radiographic change was determined using specific criteria. The maximum possible score for each foot is 26. The test compound was administered at a total daily dose of 0.1, 0.3, 1 and N3 mg/kg/day, indomethacin was administered at a total daily dose of 1 mg/kg/day, or vehicle (0.5% methylcellulose, in sterile in water) (per osb.i.d.). Started after adjuvant injection and continued for 21 days. Compounds were prepared weekly, refrigerated in the dark until use, and vortexed immediately prior to dosing.

Mouse anti-tumor model

The assay described in the article by Spranger et al. (Journal for lmmunoTherapy of Cancer 2014, 2:3) can be used to assess the synergy of a compound of the invention in combination with an effective amount of an antibody against cytotoxic T-lymphocyte antigen 4 (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.

resolve resolution

plan 1

Scenario 2:

Scenario 3:

Scenario 4

In the above schemes, the name Ar corresponds to Ar1 in formula I, and may be optionally substituted as described herein.

Example 1: 4-(1-(7-(4-(trifluoromethyl)phenoxy)-1H-indole-1-carboxamido)cyclopropyl)benzoic acid

plan 1

Step 1: Methyl 4-(1-isocyanatocyclopropyl)benzoate

To a solution of methyl 4-(1-aminocyclopropyl)benzoate (1.3 g, 6.87 mmol) and triphosgene (0.621 g, 2.09 mmol) in toluene (30 mL) was added dropwise _Et3N ( 1.45 mL, 10.46 mmol). The reaction was stirred at 0°C for 30 minutes and then at room temperature for 1 hour. The reaction was checked by TLC, the starting material was consumed by TLC, the suspension was filtered and concentrated to give 0.69 g of the title product as a pale yellow solid which was used in the next step without further purification.

Step 2: 1-Nitro-2-(4-(trifluoromethyl)phenoxy)benzene

A mixture of 1-fluoro-2-nitrobenzene (1.41 g, 10 mmol), 4-(trifluoromethyl)phenol (1.63 g, ₁₀ mmol) and _k2CO3 (2.76 g, 20 mmol) in DMF (25 ml) The solution was stirred at 100°C for 6 hours. TLC showed that all starting material was consumed and new spots were detected. 150 ml of water was added, then the mixture was extracted with EtOAc (40 ml x 3), washed with brine, dried _{over Na2SO4} and concentrated. The crude product was isolated by silica gel column chromatography (EtOAc in hexanes, from 0 to 10%) to give 2.79 g of the title product as a pale yellow solid. MS (ES-API positive): 284 (M+1)+.

Step 3: 7-(4-(Trifluoromethyl)phenoxy)-1H-indole

To a solution of 1-nitro-2-(4-(trifluoromethyl)phenoxy)benzene (2.79 g, 10 mmol) in _THF (40 mL) was added dropwise EtMgBr ( 1M, 30 mL). The resulting solution was stirred at -40°C for 1 h. TLC showed that the starting material had been consumed. 30 mL of saturated aqueous _NH4Cl was added to quench the reaction, the mixture was extracted with EtOAc (30 mL x 3), washed with brine, dried _{over Na2SO4} and concentrated. The crude product was purified by silica gel column (EtOAc in hexanes, from 0 to 7%) to give 0.91 g of the title product as a yellow solid.

MS (ES-API positive): 278 (M+1)+.

Step 4: Methyl 4-(1-(7-(4-(trifluoromethyl)phenoxy)-1H-indole-1-carboxamido)cyclopropyl)benzoate

To a solution of 7-(4-(trifluoromethyl)phenoxy)-1H-indole (0.1 g, 0.36 mmol) in THF (3 mL) at 0 °C was added n-butyllithium (1.6 M, 0.24mL). After the reaction mixture was stirred at 0°C for 15 minutes and at room temperature for 10 minutes, methyl 4-(1-isocyanatocyclopropyl)benzoate (0.078 g, 0.36) in 1 mL of THF was added. After stirring at room temperature for 5 minutes, a solution of saturated _NH4Cl (3 mL) and water (9 mL) was added to quench the reaction. The resulting solution was extracted with EtOAc (15 mL x 3), washed with brine, dried _{over Na2SO4} and concentrated. The crude product was purified via silica gel column (EtOAc in hexanes, from 0 to 15%) to give 0.07 g of the title product as a white solid. MS (ES-API positive): 495 (M+1)+. .

Step 5: 4-(1-(7-(4-(Trifluoromethyl)phenoxy)-1H-indole-1-carboxamido)cyclopropyl)benzoic acid

To methyl 4-(1-(7-(4-(trifluoromethyl)phenoxy)-1H-indole-1-carboxamido)cyclopropyl)benzoate (70 mg, 0.142 mmol) at room temperature ) in THF (12 mL) was added LiOH (5.1 mg, 0.213 mmol) in water (6 mL). The resulting solution was stirred at room temperature overnight. The desired MS peak was detected by LCMS and most of the starting material was retained, with about 10% of the starting material being decomposed. An additional 3.1 mg of LiOH was added and stirred for an additional 2 days. The expected MS peak was detected by LCMS as the main peak, and about 10% of the starting material and 18% of the decomposed components were retained. 5 mL of saturated _NH4Cl was added to quench the reaction. The mixture was extracted with EtOAc (15 mL x 3), washed with brine, dried _{over Na2SO4} and concentrated. The crude product was mixed with about 500 mg of silica and separated through a silica gel column (MeOH in DCM, from 0 to 2%) to give 0.008 g of the final product as a white solid.

¹ H NMR (400 MHz, CDCl ₃ ): 8.61 (s, 1H), 8.03 (d, 1H), 7.87 (d, 2H), 7.57 (d, 2H), 7.51 (d, 1H), 7.24 (t, 1H) ), 7.14(d, 2H), 7.01(d, 2H), 6.89(d, 1H), 6.68(d, 1H), 1.25-1.36, (m, 4H).

MS (ES-API positive): 481 (M+1)+.

Example 2: 4-(1-(7-(4-(trifluoromethyl)phenoxy)indoline-1-carboxamido)cyclopropyl)benzoic acid

Scenario 2

Step 1: 7-(4-(Trifluoromethyl)phenoxy)indoline

To a solution of 7-(4-(trifluoromethyl)phenoxy)-1H-indole (0.1 g, 0.361 mmol) in HOAc (2 mL) was added _NaBH3CN (0.045 g, 0.722 mmol) at room temperature ). The reaction mixture was stirred at room temperature overnight. TLC showed that all starting material was consumed and a new spot was detected. _The reaction was quenched with saturated _Na2CO3 and the resulting solution was extracted with EtOAc (10 mL x 3), washed with brine, dried _{over Na2SO4} and concentrated to give 0.1 g of the title compound as a pale yellow solid without further purification can be used for the next step.

MS (ES-API positive): 280 (M+1)+.

Step 2: Methyl 4-(1-(7-(4-(trifluoromethyl)phenoxy)indoline-1-carboxamido)cyclopropyl)benzoate

To 7-(4-(trifluoromethyl)phenoxy)indoline (0.1 g, 0.36 mmol) and methyl 4-(1-isocyanatocyclopropyl)benzoate (0.156 g, 0.716 mmol) ) in DCM (3 mL) was added Et3N (0.11 g, 1.07 mmol) at 0 °C. The reaction mixture was stirred at room temperature overnight. The desired MS peak was detected by LCMS and worked well. Saturated _NH4Cl solution was added to quench the reaction, and the resulting solution was extracted with EtOAc (20 mL x 3), washed with brine, dried _{over Na2SO4} and concentrated. The crude product was isolated via a silica gel column (EtOAc in hexanes, from 0 to 30%) to give 0.14 g of the final product as a pale yellow solid.

MS (ES-API positive): 497 (M+1)+.

Step 3: 4-(1-(7-(4-(Trifluoromethyl)phenoxy)indoline-1-carboxamido)cyclopropyl)benzoic acid

To methyl 4-(1-(7-(4-(trifluoromethyl)phenoxy)indoline-1-carboxamido)cyclopropyl)benzoate (0.14 g, 0.282 mmol) in THF /MeOH (1/1, 20 mL), at room temperature, was added a solution of _LiOH.H2O (0.036 g, 0.845 mmol) in water (10 mL). The resulting solution was stirred at room temperature for 5 h, the desired MS peak was detected by LCMS and most of the starting material was retained. Then another 36 mg of _LiOH.H2O was added and stirred at room temperature overnight. LCMS showed the reaction was complete. The reaction was terminated by adding 1N HCl solution, the pH value was adjusted to about 5-6, and then the organic solvent was removed under reduced pressure to obtain a suspension. The solid was filtered, washed with water and dried by an oil pump to give 0.101 g of product as a white solid.

¹ H NMR (400 MHz, CDCl ₃ ): 7.87(d, 2H), 7.55(d, 2H), 7.18(d, 1H), 7.14(d, 2H), 7.09(t, 1H), 6.99(s, 1H) ), 6.90 (t, 3H), 4.28 (t, 2H), 3.09 (t, 2H), 1.27-1.36 (m, 4H).

MS (ES-API positive): 483 (M+1)+.

Example 3: 4-((7-(4-(trifluoromethyl)phenoxy)indoline-1-carboxamido)methyl)benzoic acid

Scenario 3

Step 1: 4-Nitrophenyl 7-(4-(trifluoromethyl)phenoxy)indoline-1-carboxylate

To 7-(4-(trifluoromethyl)phenoxy)indoline (0.1 g, 0.358 mmol) and 4-nitrophenylcarbochloride (0.072 g, 0.358 mmol) in THF (0.072 g, 0.358 mmol) at 0 °C To the solution in 2 mL) was added pyridine (0.030 g, 0.358 mmol). The resulting solution was stirred at room temperature under _N2 atmosphere for 5 h. TLC showed that the starting material had been consumed. Saturated NH ₄ Cl was added to quench the reaction, and the resulting solution was extracted with EA (15 mL×3), washed with brine, dried over Na ₂ SO ₄ and concentrated to give the crude title product as a pale yellow solid without further use Purification is ready for the next step.

Step 2: Methyl 4-((7-(4-(trifluoromethyl)phenoxy)indoline-1-carboxamido)methyl)benzoate

To a solution of crude 4-nitrophenyl 7-(4-(trifluoromethyl)phenoxy)indoline-1-carboxylate (0.361 mmol) in pyridine (5 mL) at room temperature, Methyl 4-(aminomethyl)benzoate hydrochloride (0.360 g, 1.79 mmol) and DBU (0.270 g, 1.79 mmol) were added. The reaction mixture was stirred at room temperature for 2 days. TLC showed that all starting material was consumed and a new spot was detected. LCMS showed that the reaction was running well. The reaction was quenched with 1 N HCl solution, and the resulting solution was extracted with EA (15 mL x 3), washed with brine, dried over Na ₂ SO ₄ and concentrated, passed through a silica gel column (EtOAc in hexanes, from 0 to 20%) ) to isolate the crude product to give 0.088 g of the title product as a white solid. MS (ES-API positive): 471 (M+1)+.

Step 3: 4-((7-(4-(Trifluoromethyl)phenoxy)indoline-1-carboxamido)methyl)benzoic acid

To methyl 4-((7-(4-(trifluoromethyl)phenoxy)indoline-1-carboxamido)methyl)benzoate (0.088 g, 0.187 mmol) in THF/MeOH (1 /1/, 12 mL) solution, at room temperature, was added a solution of LiOH.H ₂ O (0.079 g, 1.87 mmol) in water (6 mL). The resulting solution was stirred at 50 °C for 5 h. LCMS showed the reaction was complete. The reaction was terminated by adding 1N HCl solution, the pH value was adjusted to about 3-4, and then the organic solvent was removed under reduced pressure to obtain a suspension. A lot of solids appeared, then put it in the ultrasonic cleaner for 5 minutes. The solid was collected by filtration, washed with water and dried by an oil pump to give 0.054 g of the title product as a white solid.

¹ H NMR (400 MHz, DMSO-d6): 12.83(s, 1H), 7.82(d, 2H), 7.61(d, 2H), 7.39(t, 1H), 7.22(d, 2H), 7.14(d, 2H) 1H), 7.03 (t, 1H), 6.96 (d, 2H), 6.90 (d, 1H), 4.23 (d, 2H), 4.00 (t, 2H), 3.11 (t. 2H). MS (ES-API positive): 457 (M+1)+.

Example 4: 4-(1-(1-Methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)cyclopropyl)benzoic acid

Example 5: (S)-4-(1-(1-methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)ethyl)benzene Formic acid

Scenario 4

Step 1: 4-(4-(Trifluoromethyl)phenoxy)-1H-indole

1H-Indol-4-ol (3.20 g, 24 mmol), 1-fluoro-4-(trifluoromethyl)benzene (3.58 g, 21.8 mmol) and _{Cs2CO3 (} 10.66 g, 32.7 mmol) in DMSO ( 40 mL) and stirred at 150°C for 3 hours. TLC showed that most of the starting material was consumed and the desired product was detected. 300 mL of water was added to quench the reaction, and the mixture was extracted with EtOAc (70 mL x 3), washed with brine, dried over Na ₂ SO ₄ and concentrated. The crude product was purified by silica gel column (EtOAc in hexanes, from 0 to 15%) to give 0.95 g of the title product as a brown solid. .

MS (ES-API positive): 278 (M+1)+.

Step 2: 1-Methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole

To a solution of 4-(4-(trifluoromethyl)phenoxy)-1H-indole (0.30 g, 3.047 mmol) in THF (5 mL) at 0 °C under nitrogen atmosphere was added NaH (0.062 g, 1.6 mmol, 60%). The resulting solution was stirred at 0°C for 20 minutes, then Mel (0.08 mL, 1.3 mmol) was added and stirred at room temperature for 1 h. TLC showed that the reaction went well. Saturated _NH4Cl solution was added to quench the reaction, and the resulting solution was extracted with EtOAc (10 mL x 3), washed with brine, dried _{over Na2SO4} and concentrated to give 0.315 g of the title product as a brown oil, which was Used in the next step without further purification.

MS (ES-API positive): 292 (M+1)+.

Step 3: 1-Methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxylic acid

To a solution of ₁ -methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole (0.295 g, 1.013 mmol) in DCM (5 mL) at 0 °C under N atmosphere To the solution was added TFAA (0.319 g, 1.52 mmol). The resulting solution was stirred at 0°C for 20 minutes and then at room temperature for an additional hour. LCMS showed that the reaction went well and all starting material was converted to intermediate. The solvent was removed under reduced pressure. The resulting crude product was used in the next step without further purification. To this crude product in MeOH (3 mL) was added a solution of NaOH (10 M, 3 mL). The mixture was stirred at 90°C for 3 hours. LCMS showed the reaction was complete. The reaction was cooled to room temperature and HCl solution (10%) was added to adjust pH=1-2. The resulting solution was extracted with EA (20 mL x 3), washed with brine, dried _{over Na2SO4} and concentrated to give 0.29 g of the title product as a white solid.

MS (ES-API positive): 336 (M+1)+.

Step 4: Methyl 4-(1-(1-methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)cyclopropyl)benzoate

To 1-methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxylic acid (0.032 g, 0.095 mmol), 4-(1-aminocyclopropane) at room temperature (0.022 g, 0.114 mmol) and HATU (0.054 g, 0.143 mmol) in DMF (3 mL) was added NMM (0.019 g, 0.19 mmol). The resulting solution was stirred at room temperature for 3 h. The desired MS peak was detected by LCMS, but most of the starting material was retained. Another 10 equivalents of B were added and stirred at 60°C for 3 days. LCMS showed the reaction was complete. The reaction was quenched by addition of 1N HCl solution and adjusted to pH=3-4. The resulting solution was then extracted with EtOAc (10 mL x 3), washed with brine, dried _{over Na2SO4} and concentrated. The crude product was isolated by silica gel column (MeOH in DCM, 0 to 3%) to give 0.040 g of the title product as a pale yellow solid.

MS (ES-API positive): 509 (M+1)+.

Step 5: 4-(1-(1-Methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)cyclopropyl)benzoic acid

To methyl 4-(1-(1-methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)cyclopropyl)benzoate at room temperature (0.40 g, 0.079 mmol) in THF/MeOH (1/1/, 6 mL) was added a solution of _LiOH.H2O (0.033 g, 0.790 mmol) in water (3 mL). The resulting solution was stirred at room temperature overnight. LCMS showed the reaction was complete. The reaction was terminated by adding 1N HCl solution, the pH value was adjusted to about 3-4, and then the organic solvent was removed under reduced pressure to obtain a suspension. The mixture was extracted with EtOAc (15 mL x 3), washed with brine and concentrated. The crude product was dissolved with 1.5 mL of _CH3CN and placed in an ultrasonic cleaner for 5 minutes. The solid was collected by filtration and dried by an oil pump to give 13.7 mg of the title product as a white solid.

¹ H NMR (400 MHz, DMSO-d6): 8.48 (brs, 1H), 8.00 (s, 1H), 7.80 (d, 2H), 7.72 (d, 2H), 7.47 (d, 1H), 7.32 (t, 1H), 7.23 (d, 2H), 7.18 (d, 2H), 6.85 (d, 1H), 3.98 (s, 3H), 1.20-1.29 (m, 4H).

MS (ES-API positive): 495 (M+1)+.

Step 6: (S)-4-(1-(1-Methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)ethyl)benzoic acid methyl ester

To 1-methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxylic acid (0.055 g, 0.164 mmol), methyl(S)-4- To a solution of (1-aminoethyl)benzoate (0.147 g, 0.820 mmol) and HATU (0.125 g, 0.328 mmol) in DMF (3 mL) was added NMM (0.050 g, 0.492 mmol). The resulting solution was stirred at 60°C overnight. LCMS showed that the reaction went well and all starting material was consumed. The reaction was quenched by addition of 1N HCl solution and adjusted to pH=3-4. The resulting solution was then extracted with EtOAc (10 mL x 3), washed with brine, dried _{over Na2SO4} and concentrated. The crude product was isolated by silica gel column (MeOH in DCM, 0 to 3%) to give 0.062 g of the title product as a pale yellow solid. .

MS (ES-API positive): 497 (M+1)+.

Step 7: (S)-4-(1-(1-Methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)ethyl)benzoic acid

To (S)-4-(1-(1-methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)ethyl)benzene at room temperature To a solution of methyl formate (0.062 g, 0.125 mmol) in THF/MeOH (1/1/, 10 mL) was added a solution of _LiOH.H2O (0.053 g, 1.25 mmol) in water (5 mL). The resulting solution was stirred at 50 °C for 5 h. LCMS showed the reaction was complete. The reaction was terminated by adding 1N HCl solution, the pH value was adjusted to about 5-6, and then the organic solvent was removed under reduced pressure to obtain a suspension. The mixture was extracted with EA (15 mL x 3), washed with brine and concentrated. The crude product was dissolved with 1.5 mL of _CH3CN and placed in an ultrasonic cleaner for 5 minutes. The solid was collected by filtration and washed with 1 mL of _CH3CN , then dried by an oil pump to give 21.9 mg of the title product as a white solid.

¹ H NMR (400 MHz, DMSO-d6): 12.79(s, 1H), 8.23(d, 1H), 7.97(s, 1H), 7.75(d, 2H), 7.68(d, 2H), 7.45(d, 7.45(d, 2H) 1H), 7.27(t, 1H), 7.25(d, 2H), 7.08(d, 2H), 6.78(d, 1H), 5.01(dt, 1H), 3.87(s, 3H), 1.29(d, 3H) ).

MS (ES-API positive): 483 (M+1)+.

Example 6: 4-(1-(4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)cyclopropyl)benzoic acid

Scenario 5

Step 1: 4-(4-(Trifluoromethyl)phenoxy)-1H-indole-3-carbaldehyde

To a solution of DMF (2 mL) was added _POCl3 (0.10 mL, 1.08 mmol) at 0 °C. The resulting solution was stirred at 0°C for 20 minutes and at room temperature for 20 minutes. Then 4-(4-(trifluoromethyl)phenoxy)-lH-indole (0.20 g, 0.72 mmol) in DMF (2 mL) was added and the mixture was stirred at room temperature for 2 h. TLC showed that all starting material was consumed. The desired MS peak was detected by LCMS. The mixture was poured into ₂₅ mL of saturated _Na2CO3 solution and stirred for an additional 5 minutes. The resulting solution was extracted with EA (20 mL x 3), washed with brine, dried _{over Na2SO4} and concentrated. The crude product was isolated via a silica gel column (EtOAc in hexanes, from 0 to 20%) to give 0.11 g of the title product as a white solid.

MS (ES-API positive): 306 (M+1)+.

Step 2: 4-(4-(Trifluoromethyl)phenoxy)-1H-indole-3-carboxylic acid

To a solution of 4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carbaldehyde (0.11 g, 0.361 mmol) in acetone ( ₃ mL) was added KMnO4 (0.11 g) at room temperature , 0.721 mmol) in water (1.5 mL). The resulting solution was stirred at room temperature for 4 h. The desired MS peak was detected by LCMS and the reaction ran well. 0.2 mL H2O2 _{( 10} %) was added to quench the reaction and filtered. Acetone was then removed, then the pH was adjusted to 1-2 and 40 mL of EtOAc was added, washed with brine, dried _{over Na2SO4} and concentrated. The crude product was isolated by silica gel column (MeOH in DCM, from 0 to 6%) to give 0.073 g of the title product as a brown solid.

MS (ES-API positive): 322 (M+1)+.

Step 3: Methyl 4-(1-(4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)cyclopropyl)benzoate

] at room temperature, to 1-methyl-4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxylic acid (0.073 g, 0.227 mmol), 4-(1-amino ring Propyl)methyl benzoate (0.217 g, 1.14 mmol) and HATU (0.173 g, 0.454 mmol) in DMF (3 mL) was added NMM (0.046 g, 0.454 mmol). The resulting solution was stirred at 50°C for 5 hours. LCMS showed the reaction was complete. The reaction was quenched by addition of 1N HCl solution and adjusted to pH=3-4. The resulting solution was then extracted with EtOAc (10 mL x 3), washed with brine, dried over Na ₂ SO ₄ and concentrated. The crude product was isolated via a silica gel column (MeOH in DCM, 0 to 3%) to give 0.079 g of the title product as a pale yellow solid.

MS (ES-API positive): 495 (M+1)+.

Step 4: 4-(1-(4-(4-(Trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)cyclopropyl)benzoic acid

To methyl 4-(1-(4-(4-(trifluoromethyl)phenoxy)-1H-indole-3-carboxamido)cyclopropyl)benzoate (0.079 g, 0.160 g) at room temperature mmol) in THF/MeOH (1/1, 6 mL) was added a solution of _LiOH.H2O (0.062 g, 1.60 mmol) in water (6 mL). The resulting solution was stirred at 50 °C for 5 h. LCMS showed the reaction was complete. The reaction was quenched by the addition of 1N HCl solution and the pH was adjusted to about 3-4, then the organic solvent was removed under reduced pressure to give a suspension. The mixture was extracted with EA (15 mL x 3), washed with brine and concentrated. The crude product was isolated by reverse phase column ( _CH3CN in _H2O (0.1% HCOOH) from 0 to 70%) and lyophilized to give 0.0415 g of the title product as a white solid. ¹ H NMR (400 MHz, DMSO-d6): 11.94(s,1H), 8.51(s,1H), 7.92(d,1H), 7.70(d,2H), 7.65(d,2H), 7.40(d,2H) 1H), 7.22 (t, 1H), 7.06 (d, 2H), 7.04 (d, 1H), 6.78 (d, 1H), 1.05-1.22 (m, 4H). MS (ES-API positive): 481 (M+1)+.

Claims (11)

1. A compound as shown in formula I or a pharmaceutically acceptable salt thereof,

wherein:

a, B and C' are each independently selected from N, CH and C (R);

-D-E-F-is-C (R)^c)₂-N＝C-，-N(R^c)-C(R^c)＝C-，-S-N＝C-，-O-N＝C-，-N(R^c)-N＝C-，-C(R^c)₂-C(R^c)₂-N-，-C(R^c)＝C(R^c)-N-，-C(R^c)＝N-N-，-N＝N-N-，-O-C(R^c) -C-, or-S-C (R)^c)＝C-；

R^1aAnd R^2aEach independently is H, C_1-6Alkyl radical, C_1-6Cycloalkyl radical, C_1-6Fluorocycloalkyl group, or C_1-6A fluoroalkyl group; or R¹And R²Together with the carbon atom to which they are both attached form a three-to six-membered carbocyclic ring, which carbocyclic ring is optionally substituted by R^cSubstituted or formed into a three-to six-membered ring containing one or two heteroatoms, each independently S, S (O)₂S (O), (NH), O, or NR^eWherein R is^eIs H, C_1-6Alkyl radical, C_1-6Cycloalkyl radical, C_1-6Fluorocycloalkyl group, C_1-6Fluoroalkyl, aryl, heteroaryl, C (O) C_1-6Alkyl, C (O) aryl, S (O)₂Alkyl, or S (O)₂An aryl group;

each R^cIndependently of one another is H, halogen, C_1-4Alkyl radical, C_1-4Fluoroalkyl radicals, C_1-4Alkoxy radical, C_1-4Fluoroalkoxy, or acetyl;

each R is independently H, C_1-4Alkyl radical, C_1-4Fluoroalkyl radicals, C_1-4Alkoxy radical, C_1-4Fluoroalkoxy, or acetyl;

y is-CH₂-, -C (O) -, -C (S) -, or-S (O)₂-；

X is CH₂O, or S;

w is NH or O;

Ar¹and Ar²Each independently is C_3-6Cycloalkyl, aryl, heteroaryl, or heterocyclyl, or C_3-6Fused analogues of cycloalkyl, aryl, heteroaryl or heterocyclyl, and Ar¹And Ar²Each optionally substituted by 1-3R^bSubstituted by groups;

each R^bIndependently of one another is H, halogen, C_1-6Alkyl radical, C_1-6Alkoxy, or C_1-6A fluoroalkyl group;

R^fis-CO₂H，-CO₂M, or

And is

M is a pharmaceutically acceptable salt or ester prodrug group.

2. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula Ia

Wherein:

a, B and C' are each independently N, CH or C (R);

-D-E-F-is-C (R)^c)₂-N＝C-，-N(R^c)-C(R^c)＝C-，-S-N＝C-，-O-N＝C-，-N(R^c)-N＝C-，-C(R^c)₂-C(R^c)₂-N-，-C(R^c)＝C(R^c)-N-，-C(R^c)＝N-N-，-N＝N-N-，-O-C(R^c) -C-, or-S-C (R)^c)＝C-；

R¹And R²Each independently of the others being H, halogen, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Cycloalkyl radical, C_1-6Fluorocycloalkyl group, C_1-6Fluoroalkyl or SF₅；

Each R^bIndependently of one another is H, halogen, C_1-6Alkyl radical, C_1-6Alkoxy, or C_1-6A fluoroalkyl group;

R^1aand R^2aEach independently is H, C_1-6Alkyl radical, C_1-6Cycloalkyl radical, C_1-6Fluorocycloalkyl group, or C_1-6Fluoroalkyl radicals, or

R¹And R²Together with the carbon atom to which they are both attached form a three-to six-membered carbocyclic ring, optionally substituted with R^cSubstituted, or formed into a three-to six-membered ring containing one or two heteroatoms, each independently S, S (O)₂S (O), (NH), O, or NR^eWherein R is^eIs H, C_1-6Alkyl radical, C_1-6Cycloalkyl radical, C_1-6Fluorocycloalkyl group, C_1-6Fluoroalkyl, aryl, heteroaryl, C (O) C_1-6Alkyl, C (O) aryl, S (O)₂Alkyl or S (O)₂An aryl group;

each Rc is independently H, C_1-4Alkyl radical, C_1-4Fluoroalkyl radicals, C_1-4Alkoxy radical, C_1-4Fluoroalkoxy or acetyl;

each R is independently H, halogen, C_1-4Alkyl radical, C_1-4Fluoroalkyl radicals, C_1-4Alkoxy radical, C_1-4Fluoroalkoxy, or acetyl;

x is CH₂O or S;

each R^bIndependently of one another is H, halogen, C_1-6Alkyl radical, C_1-6Alkoxy, or C_1-6A fluoroalkyl group;

n is 0, 1, 2 or 3.

3. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is of formula Ib:

wherein:

-D-E-F-is-C (R)^c)₂-N＝C-，-N(R^c)-C(R^c)＝C-，-S-N＝C-，-O-N＝C-，-N(R^c)-N＝C-，-C(R^c)₂-C(R^c)₂-N-，-C(R^c)＝C(R^c)-N-，-C(R^c)＝N-N-，-N＝N-N-，-O-C(R^c)＝C-，-S-C(R^c)＝C-；

R¹And R²Each independently of the others being H, halogen, C_1-6Alkyl radical, C_1-6Alkoxy radical, C_1-6Cycloalkyl radical, C_1-6Fluorocycloalkyl group, C_1-6Fluoroalkyl or SF₅；

Each R^bIndependently of one another is H, halogen, C_1-6Alkyl radical, C_1-6Alkoxy, or C_1-6A fluoroalkyl group;

R^1aand R^2aEach independently is H, C_1-6Alkyl radical, C_1-6Cycloalkyl radical, C_1-6Fluorocycloalkyl group, or C_1-6A fluoroalkyl group; or R¹And R²Together with the carbon atom to which they are both attached form a three-to six-membered carbocyclic ring, which carbocyclic ring may optionally be substituted by R^cSubstituted or formed into a three-to six-membered ring containing one or two heteroatoms, each independently S, O or R^eWherein R is^eIs H or C_1-6An alkyl group;

each Rc is independently H, C_1-4Alkyl radical, C_1-4Fluoroalkyl radicals, C_1-4Alkoxy radical, C_1-4Fluoroalkoxy or acetyl;

each R is independently H, halogen, C_1-4Alkyl radical, C_1-4Fluoroalkyl radicals, C_1-4Alkoxy radical, C_1-4Fluoroalkoxy or acetyl;

each Rb is independently H, halogen, C_1-6Alkyl radical, C_1-6Alkoxy, or C_1-6A fluoroalkyl group; and

n is 0, 1, 2 or 3.

4. The compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the fused ring moiety in formula I

A heterocyclic moiety selected from:

5. the compound of claim 1, or a pharmaceutically acceptable salt thereof, wherein the compound is:

4- (1- (7- (4- (trifluoromethyl) phenoxy) -1H-indole-1-carboxamide) cyclopropyl) benzoic acid;

4- (1- (7- (3- (trifluoromethyl) phenoxy) -1H-indole-1-carboxamide) cyclopropyl) benzoic acid;

4- (2- (7- (3- (trifluoromethyl) phenoxy) -1H-indole-1-carboxamide) propan-2-yl) benzoic acid;

(±)4- (1- (7- (3- (trifluoromethyl) phenoxy) -1H-indole-1-carboxamide) ethyl) benzoic acid;

(±)4- (1- (7- (4- (trifluoromethyl) phenoxy) -1H-indole-1-carboxamide) ethyl) benzoic acid;

(±)4- (1- (4- (4- (trifluoromethyl) phenoxy) -1H-indole-3-carboxamide) ethyl) benzoic acid;

4- (1- (4- (4- (trifluoromethyl) phenoxy) -1H-indole-3-carboxamide) cyclopropyl) benzoic acid;

4- (1- (4- (3- (trifluoromethyl) phenoxy) -1H-indole-3-carboxamide) cyclopropyl) benzoic acid;

4- (1- (4- (3- (trifluoromethyl) phenoxy) benzofuran-3-carboxamido) cyclopropyl) benzoic acid;

4- (1- (4- (4- (trifluoromethyl) phenoxy) benzofuran-3-carboxamido) cyclopropyl) benzoic acid;

(±)4- (1- (4- (4- (trifluoromethyl) phenoxy) benzofuran-3-carboxamide) ethyl) benzoic acid;

(±)4- (1- (4- (4- (trifluoromethyl) phenoxy) benzo [ b ] thiophene-3-carboxamide) ethyl) benzoic acid;

(±)4- (1- (4- (3- (trifluoromethyl) phenoxy) benzo [ b ] thiophene-3-carboxamide) ethyl) benzoic acid;

4- (1- (4- (3- (trifluoromethyl) phenoxy) benzo [ b ] thiophene-3-carboxamide) cyclopropyl) benzoic acid;

4- (1- (4- (4- (trifluoromethyl) phenoxy) benzo [ b ] thiophene-3-carboxamide) cyclopropyl) benzoic acid;

4- (1- (4- (4- (trifluoromethyl) phenoxy) benzo [ d ] isothiazol-3-carboxamide) cyclopropyl) benzoic acid;

4- (1- (4- (3- (trifluoromethyl) phenoxy) benzo [ d ] isothiazole-3-carboxamide) cyclopropyl) benzoic acid

(±)4- (1- (4- (3- (trifluoromethyl) phenoxy) benzo [ d ] isothiazole-3-carboxamide) ethyl) benzoic acid;

(±)4- (1- (4- (4- (trifluoromethyl) phenoxy) benzo [ d ] isothiazole-3-carboxamide) ethyl) benzoic acid;

(±)4- (1- (4- (4- (trifluoromethyl) phenoxy) benzo [ d ] isoxazole-3-carboxamide) ethyl) benzoic acid;

(±)4- (1- (4- (3- (trifluoromethyl) phenoxy) benzo [ d ] isoxazole-3-carboxamide) ethyl) benzoic acid;

4- (1- (4- (3- (trifluoromethyl) phenoxy) benzo [ d ] isoxazole-3-carboxamido) cyclopropyl) benzoic acid;

4- (1- (4- (4- (trifluoromethyl) phenoxy) benzo [ d ] isoxazole-3-carboxamido) cyclopropyl) benzoic acid;

4- (1- (7- (4- (trifluoromethyl) phenoxy) indoline-1-carboxamide) cyclopropyl) benzoic acid; or

4- (1- (7- (3- (trifluoromethyl) phenoxy) indolin-1-carboxamido) cyclopropyl) benzoic acid.

6. A pharmaceutical composition comprising a compound of any one of claims 1-5 and a pharmaceutically acceptable carrier.

7. A method of treating a subject having a disorder mediated by the action of PGE2 at EP4 receptors, comprising administering to a subject in need thereof an effective amount of a compound of any one of claims 1-5 or a pharmaceutical composition of claim 6.

8. The method of claim 7, wherein the disease is an inflammatory disease or cancer.

9. The method of claim 8, wherein the inflammatory disease is arthritis, acne vulgaris, asthma, an autoimmune disease, an autoinflammatory disease, an celiac disease, chronic prostatitis, colitis, diverticulitis, glomerulonephritis, hidradenitis suppurativa, allergy, inflammatory bowel disease, interstitial cystitis, mast cell activation syndrome, mastocytosis, otitis media, pelvic inflammatory disease, reperfusion injury, rheumatic fever, rheumatoid arthritis, rhinitis, sarcoidosis or vasculitis.

10. The method of claim 7, wherein the disorder is cancer and the method comprises administering to a subject in need thereof an effective amount of a compound of any one of claims 1-5 in combination with a second therapeutic agent that is an antibody, a kinase inhibitor, an IDO inhibitor, a TDO inhibitor, a STING activator, an HDAC inhibitor or a chemotherapeutic agent.

11. The method of claim 10, wherein the antibody is an antibody against CTCLA4, PDL1, or PD 1.