阅读说明：本技术 作为前列腺素e2(pge2)受体调节剂的新型n-苄基-2-苯氧基苯甲酰胺衍生物 (Novel N-benzyl-2-phenoxybenzamide derivatives as prostaglandin E2(PGE2) receptor modulators ) 是由 J·C·帕洛米诺拉里亚 J·卡马乔戈麦斯 R·罗德里格斯伊格莱西亚斯 I·韦利利亚马丁内斯 于 2020-02-07 设计创作，主要内容包括：本发明涉及作为前列腺素E2(PGE2)的EP4和/或EP2受体的调节剂的式(I)的新型的、任选地取代的N-苄基-2-苯氧基苯甲酰胺衍生物,其制备方法,包括所述化合物的药物组合物和所述化合物用于治疗可通过调节前列腺素E2(PGE2)的EP4和/或EP2受体而改善的病理病症、紊乱或疾病比如癌症疾病、疼痛、炎症、神经退行性疾病和肾脏疾病。(The present invention relates to novel, optionally substituted N-benzyl-2-phenoxybenzamide derivatives of formula (I) as modulators of the EP4 and/or EP2 receptors of prostaglandin E2(PGE2),)

1. A compound of formula (I):

wherein:

-A represents a group selected from phenyl and a five-or six-membered heteroaryl group containing one, two or three heteroatoms selected from N, S and O,

-R¹and R²Independently represent a hydrogen atom, a halogen atom and C_1-3Radical of alkyl, or R¹And R²Together with the carbon atom to which they are attached form C_3-4A cycloalkyl group,

-R³、R⁴、R⁵and each R⁶Independently represent a hydrogen atom, a halogen atom, a cyano group, a carbamoyl group, a linear or branched C_1-3Haloalkyl, straight-chain or branched C_1-3Alkyl radical, C_3-4Cycloalkyl, straight or branched C_1-3The group of alkoxy and pyridyl,

m is an integer of 1 to 5,

and pharmaceutically acceptable salts thereof.

2. The compound of claim 1, wherein R¹、R²、R³、R⁴And R⁵Represents a hydrogen atom.

3. A compound according to any one of claims 1 to 2, wherein a represents phenyl.

4. The compound of claim 3, wherein each R⁶Independently represents a group selected from:

a) a hydrogen atom, and a nitrogen atom,

b) halogen atom

c) The cyano group(s),

d) carbamoyl radical

e) Straight or branched C_1-3A haloalkyl group.

5. A compound according to any one of claims 1 to 4, wherein A represents phenyl and each R represents⁶Independently represents a group selected from fluorine, trifluoromethyl and cyano, and m is an integer of 1 to 4.

6. A compound according to any one of claims 1 to 2, wherein a represents a five or six membered heteroaromatic ring.

7. A compound according to claim 6, wherein A represents a group selected from pyridyl or thiophenyl, each R⁶Independently represents a group selected from trifluoromethyl and cyano, and m is an integer of 1 to 4.

8. The compound of claim 1, wherein R¹、R²、R³、R⁴And R⁵Represents a hydrogen atom, A represents a phenyl group and each R⁶Independently represent a group selected from fluorine and trifluoroA methyl group, and m is an integer of 1 to 5.

9. A compound according to any preceding claim, wherein m is an integer from 1 to 2.

10. A compound according to any one of claims 1 to 2, wherein A represents a group selected from phenyl, pyridinyl, pyrimidinyl, thiophenyl, R¹、R²、R³、R⁴、R⁵Is a hydrogen atom, each R⁶Independently represents a group selected from a hydrogen atom, fluorine, chlorine, cyano, trifluoromethyl and pyridyl, and m is an integer of 1 to 2.

11. The compound of claim 1, which is one of:

4- ((2- (4-fluorophenoxy) -5- (pyridin-4-yl) benzamido) methyl) benzoic acid

4- ((4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((4 '-chloro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((4 '-fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((3 '-fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((2 '-fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((4 '-cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((3 '-cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (3-fluoropyridin-4-yl) benzamido) methyl) benzoic acid)

4- ((2- (4-fluorophenoxy) -5- (pyridin-3-yl) benzamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (3-chloropyridin-4-yl) benzamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (pyrimidin-5-yl) benzamido) methyl) benzoic acid

4- ((3',4' -difluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((4- (4-fluorophenoxy) -4'- (trifluoromethyl) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (pyridin-2-yl) benzamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (pyrimidin-4-yl) benzamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (phenylthio-2-yl) benzamido) methyl) benzoic acid

4- ((3',5' -difluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl)) benzoic acid

4- ((3 '-chloro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((3' -fluoro-4- (4-fluorophenoxy) -5' - (trifluoromethyl) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((3' -carbamoyl-5 ' -chloro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-ylcarboxamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (1H-pyrazol-4-yl) benzamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (1-methyl-1H-pyrazol-4-yl) benzamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (1-methyl-1H-pyrazol-5-yl) benzamido) methyl) benzoic acid

4- ((4- (4-fluorophenoxy) -3'- (pyridin-4-yl) - [1,1' -biphenyl ] -3-ylcarboxamido) methyl) benzoic acid

4- ((5- (5-carbamoylfuran-2-yl) -2- (4-fluorophenoxy) benzamido) methyl) benzoic acid

3-fluoro-4- ((4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl ] -3-ylcarboxamido) methyl) benzoic acid

(R) -4- (1- (4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl ] -3-carboxamido) ethyl) benzoic acid

4- (1- (4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl ] -3-carboxamido) cyclopropyl) benzoic acid

4- ((3' -fluoro-5 ' -cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((3' -chloro-5 ' -cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid,

and pharmaceutically acceptable salts thereof.

12. A compound as defined in any one of claims 1 to 11 for use in the treatment or prevention of a disease or pathological condition ameliorated by the modulation of EP4 and/or EP2 receptors of prostaglandin E2(PGE2), wherein the disease or pathological condition is selected from cancer, pain, neurodegenerative diseases, osteoarthritis, rheumatoid arthritis, endometriosis and renal disease.

13. A compound as defined in any one of claims 1 to 11 for use in the treatment or prevention of cancer, in particular a type of cancer suitable for treatment with immunotherapy.

14. Use of a compound according to any one of claims 1 to 11 in the manufacture of a medicament for the treatment or prevention of a disease or pathological condition ameliorated by the modulation of EP4 and/or EP2 receptors of prostaglandin E2(PGE2), wherein the disease or pathological condition is selected from cancer, pain, neurodegenerative diseases, osteoarthritis, rheumatoid arthritis, endometriosis and renal disease.

15. Use of a compound according to any one of claims 1 to 11 in the manufacture of a medicament for the treatment or prophylaxis of cancer, in particular of a type of cancer suitable for treatment with immunotherapy.

16. A method of treating or preventing a disease or pathological condition ameliorated by the modulation of EP4 and/or EP2 receptors of prostaglandin E2(PGE2), wherein the disease or pathological condition is selected from the group consisting of cancer, pain, neurodegenerative diseases, osteoarthritis, rheumatoid arthritis, endometriosis and kidney disease, comprising administering to a subject in need thereof an effective amount of a compound according to any one of claims 1 to 11.

17. A method of treating or preventing cancer, in particular a type of cancer suitable for treatment with immunotherapy, comprising administering to a subject in need thereof an effective amount of a compound according to any one of claims 1 to 11.

18. A pharmaceutical composition comprising a compound according to any one of claims 1 to 11, a pharmaceutically acceptable diluent or carrier and optionally a therapeutically effective amount of other chemotherapeutic agents, anti-inflammatory agents, steroids, immunotherapeutic agents and other agents such as therapeutic antibodies.

19. A combination product comprising a compound according to any one of claims 1 to 11, or a pharmaceutically acceptable salt thereof, and at least one therapeutic agent selected from chemotherapeutic agents, anti-inflammatory agents, steroids, immunosuppressive agents, immunotherapeutic agents, therapeutic antibodies and EP4 and/or EP2 modulators, in particular those selected from: anti-CTLA 4 antibody selected from ipilimumab and tremelimumab, anti-PD 1 antibody such as nivolumab, pembrolizumab, cimepriazumab, pirlizumab, spartalizumab, MEDI0680, REGN2810, and AMP-224, anti-PDL 1 antibody such as atelizumab, avizumab, duvacizumab, and MDX-1105; carboplatin, carmustine (BCNU), cisplatin, cyclophosphamide, etoposide, irinotecan, lomustine (CCNU), methotrexate, procarbazine, temozolomide, vincristine.

Technical Field

The present invention relates to novel, optionally substituted N-benzyl-2-phenoxybenzamide derivatives as modulators of the EP4 and/or EP2 receptors for prostaglandin E2(PGE 2).

Other objects of the present invention are to provide processes for preparing these compounds; pharmaceutical compositions comprising an effective amount of these compounds, and the use of said compounds for the treatment of pathological conditions or diseases which can be ameliorated by modulators of the EP4 and/or EP2 receptors, such as cancer diseases, pain and inflammatory conditions, such as acute and chronic pain, osteoarthritis, rheumatoid arthritis disease, endometriosis and kidney disease.

Background

Prostaglandin E2(PGE2) is involved in a variety of biological processes such as pain, fever, vascular tone regulation, renal function, mucosal integrity, inflammation, angiogenesis and tumor growth. PGE2 often causes complex and diverse effects, which can be attributed to the activation of its four so-called E-type prostaglandin receptors (EP1 to EP 4). The physiological activities of PGE2 are mediated by 4G protein-coupled plasma membrane receptors, which are recognized as E prostaglandin receptors 1-4(EP1, EP2, EP3 and EP4), each of which can activate different downstream signaling pathways. (SURIMOTO, Yukihiko; NARUMIYA, Shuh. prostaglandin E receivers. journal of Biological Chemistry,2007, vol.282, no 16, p.11613-11617).

PGE2 is the major eicosanoid detected under inflammatory conditions and, in addition, it is involved in various physiological and/or pathological conditions such as hyperalgesia, uterine contractions, digestive peristalsis, wakefulness, inhibition of gastric acid secretion, blood pressure, platelet function, bone metabolism, angiogenesis, cancer metastasis and the like. Eicosanoids: (From Biotechnology to Therapeutic Applications, Folco, Samuelsson, Maclouf and Velo eds., Plenum Press, New York,1996, Chapter 14, p.137-154); (Journal of Lipid Mediators and Cell Signalling,14:83-87(1996)), (Prostagladins and Other Lipid Mediators,69:557-573 (2002)).

On the other hand, PGE2 was reported to be highly expressed in cancer tissues of different types of cancers, and PGE2 was also elucidated to be associated with the development of cancers and disease conditions. PGE2 is known to be involved in activating cell proliferation and inhibiting cell death (apoptosis) and plays an important role in cancer progression and metastasis.

Each EP receptor subtype has a specific distribution in humans; EP 1: myometrium, pulmonary vein, colon, skin, mast cells; EP 2: white blood cells, smooth muscle, Central Nervous System (CNS), reproductive system, bone; EP 3: central nervous system, cardiovascular system, reproductive system, kidney, bladder; EP 4: leukocytes, smooth muscle, cardiovascular system and skeleton (SUGIMOTO, Yukihiko; NARUMIYA, Shuh. journal of Biological Chemistry,2007, vol.282, no 16, p.11613-11617) and (Woodward, D.F. et al, (2011) International Union of Basic and Clinical pharmacology.LXXXIII: Pharmacol.Rev.63, 471-5385).

EP2 receptor

Studies using EP2 receptor knockout mice have demonstrated the role of EP2 receptor in malignancy, and EP2 receptor deficient mice have significantly reduced lung, skin and breast tumors after exposure to carcinogenic promoters. Genetic ablation of the EP2 receptor also reduced the size and number of intestinal polyps in Adenomatous Polyposis (APC)1309 mice, which were genetically susceptible to intestinal polyp development. Furthermore, the EP2 receptor has been shown to be expressed by tumor cells in a variety of cancers including colon, prostate and breast cancers (GUSTAFSSON, Annika et al International journal of cancer,2007, vol.121, no 2, p.232-240) and (Chang SH et al, (2004) Proc Natl Acad Sci U S A101: 591-.

In prostate cancer, Androgen Deprivation Therapy (ADT) is well known as a major treatment, but the disease frequently recurs and becomes castration resistant in almost all patients. Mechanistically, it has been determined that elevated PGE2-EP2 signaling inhibits CD4 expression in thymocytes following androgen deprivation. Therapeutically, inactivation of PGE2 signal with celecoxib significantly inhibited the onset of castration-resistant prostate cancer (CRPC). These results indicate a new therapeutic approach to treat prostate cancer combining ADT with PGE2 inhibition (WANG, c. et al, Cell research, 2018).

PGE2-EP2 signaling as a node of chronic inflammation, shapes the tumor microenvironment, and is therefore a powerful candidate target for colorectal cancer chemoprevention (AOKI, Tomohiro; NARUMIYA, shuh. inflammation and regeneration,2017, vol.37, no1, p.4.).

Furthermore, a recent study showed that PGE2 activation of the EP2 receptor can significantly enhance the invasion and migration capacity of hepatocellular carcinoma cells by up-regulating the expression level of the EMT key inducer Snail (CHENG, shann-Yu, et al oncology reports,2014, vol.31, no 5, p.2099-2106.) and (ZANG, Shengbing et al Human pathology,2017, vol.63, p.120-127). The EP2 receptor is also associated with metastasis of breast cancer, in part because it is capable of altering the response of cells to TGF-. beta.s (Tian M, Schiemann WP (2010); FASEB J24: 1105-1116).

EP2 antagonism of neurodegenerative diseases

Chronic inflammatory neurodegenerative diseases, such as epilepsy, Alzheimer's Disease (AD), Parkinson's Disease (PD), and Amyotrophic Lateral Sclerosis (ALS), result in mortality and morbidity in a significant portion of the human mouth. AD and PD are particularly associated with dementia symptoms in the elderly. However, all these diseases share a common feature, namely, the overactivation of inflammatory pathways. Interestingly, PGE2 is the major prostaglandin produced during the progression of these diseases and has been shown to mediate pro-inflammatory functions via the EP2 (and to a lesser extent EP4) receptor. Studies have shown that administration of a potent and selective EP2 antagonist TG4-15585 significantly blocks neuronal damage in the hippocampus (> 60% of the pulmonia, 80% of CA3 and > 90% of the CA1 region) in the pilocarpine status epilepticus model. (JIANG, Jianxing et al, Proceedings of the National Academy of Sciences,2012, vol.109, no 8, p.3149-3154).

Another potent EP2 antagonist, TG6-10-186, could inhibit neurodegeneration, inflammatory cytokine and chemokine upregulation and glial activation in mice following status epilepticus, suggesting that EP2 antagonists have potential therapeutic benefits in blocking epileptogenesis. (JIANG, Jianxinong et al, Proceedings of the National Academy of Sciences,2013, vol.110, no 9, p.3591-3596).

Based on the beneficial effects observed in EP2 knock-out mouse models of AD, PD and ALS (see above), EP2 receptor antagonists are useful as adjunctive therapeutic agents for chronic inflammatory neurodegenerative diseases including epilepsy, alzheimer's disease, parkinson's disease and amyotrophic lateral sclerosis. (LIANG, Xibin et al, Annals of Neurology: Official Journal of the American Neurology Association and the Child Neurology Society,2008, vol.64, no 3, p.304-314) ((Liang X et al, J.Neurosci.2005; 25: 10180-; (Jin J et al, Journal of neuroingmigration.2007).

EP4 receptor

Unlike the EP2 receptor, the EP4 receptor is expressed in a variety of tissues and cells including immune, osteoarticular, cardiovascular, gastrointestinal and respiratory systems, and cancer cells. Recent findings suggest that modulation of EP4 signaling may be involved in therapeutic strategies for colon Cancer (MUTOH, Michihiro et al, Cancer research,2002, vol.62, no1, p.28-32.), aortic aneurysms (YOKOYAMA, Utako et al, PloS one,2012, vol.7, no 5, p.e36724.), rheumatoid arthritis (chenn, q. et al, British joural of pharmacology,2010, vol.160, no 2, p.292-310.), osteoporosis and autoimmune diseases (YAO, Chengcan et al, Nature media, 2009, vol.15, no 6, p.633.).

Thus, modulation of EP4 signaling has received greater attention as a potential therapeutic target.

Recent reports have shown that EP4 receptors expressed in certain types of cancer promote proliferation and metastasis of tumor cells (YOKOYAMA et al, Pharmacological reviews,2013, vol.65, no 3, p.1010-1052).

In breast cancer, studies have shown that tumor progression and metastasis result from a variety of cellular events, including inactivation of host anti-tumor immune cells, such as Natural Killer (NK) and T cells, increased immunosuppressive function of tumor-associated macrophages, increased cellular endothelial growth factor (VEGF) -a, increased lymphangiogenesis (due to upregulation of VEGF-C/D), and stimulation of stem cells due to upregulation of a variety of angiogenic factors, promotion of tumor cell migration, invasion, and the cytologic (SLC) phenotype in tumor-associated angiogenic cancer cells is mediated primarily by activation of Prostaglandin (PG) E receptor EP4 on tumors or host cells, and selective EP4 antagonists can alleviate all of these events tested in vitro and in vivo in breast cancer mice expressing COX-2 or immunodeficient mice carrying COX-2 overexpressing human breast cancer xenografts (MAJUMDER, mousumi et al, International journal of molecular sciences,2018, vol.19, no 4, p.1019).

EP4 expression levels were also associated with prostate cancer Cell invasiveness, and the EP4 specific antagonist ONO-AE3-208 inhibited Cell invasion, migration and bone metastasis (XU, Song et al, Cell biology and biology, 2014, vol.70, no1, p.521-527).

Other studies have shown that inhibition of EP4 attenuates Renal Cell Carcinoma (RCC) intravascular and metastasis by down-regulating CD24, and that P-selectin is involved in intratumoral infiltration (intravasation), suggesting that these molecules may be therapeutic targets for late stage RCC treatment (ZHANG, Yushan et al, Cancer letters,2017, vol.391, p.50-58).

The EP4 antagonist L-161,982 induced apoptosis, cell cycle arrest and inhibition of prostaglandin E2-induced proliferation in Oral squamous carcinoma Tca8113 cells (LI, Xiaohui et al, The Journal of Oral Pathology & Medicine,2017, vol.46, no 10, p.991-997).

Other studies have shown that EP4 antagonism is a more selective inhibition of PGE2 signalling, reducing the number of Cancer Stem Cells (CSCs) in the tumour and increasing tumour chemosensitivity. EP4 antagonism converts CSCs into non-stem cells sensitive to chemotherapy by triggering extracellular vesicle/exosome release and enhances the tumor response to conventional chemotherapy (LIN, Meng-Chieh et al, International Journal of Cancer, 2018).

In addition, selective EP4 receptor antagonists are also expected to provide an attractive therapeutic approach for autoimmune diseases such as Inflammatory Bowel Disease (IBD), Rheumatoid Arthritis (RA) and Multiple Sclerosis (MS)16-18 by inhibiting interleukin 23(IL-23) production and inhibiting T helper 1(Th1) and T helper 17(Th 17).

The role of the EP4 receptor in liver cancer has recently been reported in the non-patent literature. PGE2/EP4 receptor signaling upregulates c-Myc expression by PKA/CREB activation and leads to cell growth in HCC cells in vitro (XIA, Shukai et al, Oncology reports,2014, vol.32, no 4, p.1521-1530).

EP4 receptor signaling activated by PGE2 may be involved in various pathological conditions such as pain (in particular inflammatory, neurological and visceral), inflammation, neuroprotection, dermatitis, bone disease, promotion of immune system dysfunction in sleep, renal regulation, gastric or intestinal mucus secretion and duodenal bicarbonate secretion. Studies have shown that PGE2 inhibits proteoglycan synthesis via EP4 receptors and stimulates matrix degradation in osteoarthritic chondrocytes. Targeting EP4, rather than cyclooxygenase 2, may represent a strategy for future osteoarthritis disease improvement (ATTUR, Mukundan et al, The Journal of Immunology,2008, vol.181, no 7, p.5082-5088). Other studies have shown that pharmacological blockade of the prostaglandin EP4 receptor may represent a novel therapeutic strategy for alleviating the signs and symptoms of osteoarthritis and/or rheumatoid arthritis (MURASE, Akio et al, European journal of pharmacology,2008, vol.580, no 1-2, p.116-121).

On the other hand, several studies have shown a role for the EP4 receptor in the development of kidney disease.

In this sense, recent studies demonstrated the kidney protective properties of the PGE2 EP4 receptor selective antagonist ASP7657 in 5/6 nephrectomized rats, a model of Chronic Kidney Disease (CKD). This study showed that ASP7657 inhibited the progression of chronic renal failure by modulating renin release and improving renal hemodynamics (ELBERG, Dorit et al, prostagladins & other lipid mediators,2012, vol.98, no 1-2, p.11-16).

Other studies disclose that EP4 antagonists may improve the nephrotoxic serum nephritis NTS phenotype primarily by reducing Cxcl-5 production in the tubule cells and thus reducing the infiltration of nephroneutrophils (ARINGER, Ida et al, American Journal of Physiology-crude Physiology, 2018).

In rodent models of diabetic and non-diabetic Chronic Kidney Disease (CKD), EP4 inhibition reduces kidney damage by mechanisms other than broad-spectrum COX inhibition or "standard-of-care" blockade of the renin angiotensin system (THEIME, Karina et al, Scientific Reports,2017, vol.7, no1, p.3442.)

Finally, other studies mention that EP4 plays a key role in anti-inflammatory and anti-apoptotic effects through Akt and NF- κ B signaling after paricalcitol pretreatment in Lipopolysaccharide (LPS) -induced renal proximal tubule cell injury (HONG, Yu Ah et al, trunk research and clinical practice,2017, vol.36, no 2, p.145).

It is well known that prostaglandin E2(PGE2) contributes to cyst development in a genetically non-orthologous model of Autosomal Dominant Polycystic Kidney Disease (ADPKD). PGE2 activates aberrant signaling pathways in renal epithelial cells that lack polycystic protein 1(PC-1), which contribute to the proliferative and secretory phenotypic characteristics of ADPKD. Antagonism of the EP4 receptor restored the growth advantage of Renal epithelial cells lacking polycystic protein-1 (PC-1 deficient cells), suggesting a central role for the EP4 receptor in proliferation (LIU, Yu et al, American Journal of Physiology-crude Physiology,2012, vol.303, no 10, p.f 1425-F1434).

Prostaglandin E2(PGE2) stimulates cAMP formation and cyst formation in human ADPKD cells. Studies in isolated human ADPKD renal epithelial cells have shown that the effects of PGE2 appear to be mediated by the EP2 receptor, suggesting that EP2 receptor antagonists have therapeutic potential in treating cysts of ADPKD. Prostaglandin E receptor selective antagonists that inhibit the activity of EP2 or EP4 may be used as pharmacological strategies to limit cyst formation and ADPKD progression (ELBERG, Dorit et al, prostagladins & other lipid mediators,2012, vol.98, no 1-2, p.11-16).

EP2 and EP4 receptor modulation are also associated with a variety of diseases.

Activation of the PGE2/EP2 and PGE2/EP4 signaling pathways positively modulate PD-1 levels in infiltrating CD8+ T cells in lung cancer patients, creating immune tolerance in the lung cancer microenvironment (WANG, Jinhong et al, Oncology letters,2018, vol.15, no1, p.552-558).

EP2/EP4 activation is associated with induction of urothelial cancer initiation and growth and chemoresistance, presumably via down-regulation of phosphatase and tensin homolog (PTEN) expression. In the BC line, EP2/EP4 antagonists and celecoxib effectively inhibited cell viability and migration, as well as enhanced PTEN expression (KASHIWAGI, Eiji et al, British journal of cancer,2018, vol.118, no 2, p.213).

It is well known that upregulation of the S1P3 receptor in metastatic breast cancer cells increases migration and invasion by inducing PGE2 and EP2/EP4 activation. (FILIPENKO, Iuliia et al, Biochimica et al, biophysica act (BBA) -molecular and cell biology of lipids,2016, vol.1861, no11, p.1840-1851).

Human prostate Cancer is also known to be associated with EP4 and EP2 overexpression and with reduced expression of EP3 (HUANG, Hosea FS et al, signalic of direct expression of prostaglandin EP4 and EP3 receptors in human pro-state Cancer Research,2013, p.molcanes.0464.2012). Likewise, other studies have shown that prostaglandin E2 modulates tumor angiogenesis in prostate cancer via prostaglandin E2(EP2) mediated and EP4 mediated pathways. (JAIN, Shalini et al, Cancer Research,2008, vol.68, no 19, p.7750-7759).

Other studies suggest that PGE2 promotes renal cancer cell invasion by increasing SN12C cell invasion through a signaling pathway comprising EP2 and EP 4. (LI, ZHENyu et al, PGE2 proteins cell innovative varied ralA. Oncogene,2013, vol.32, no11, p.1408) and (LI, ZHENyu et al, Oncogene,2013, vol.32, no11, p.1408).

Agonists of the PGE2 receptor (EP2 and EP4) promote the migration of melanoma cells, while antagonists of the PGE2 receptor inhibit the ability of melanoma cells, a highly invasive skin cancer. (VAID, Mudit et al, American journel of cancer research,2015, vol.5, no11, p.3325).

Furthermore, there has been evidence that inhibition of EP2 and EP4 indicate that PGE2 induces COX-2 protein expression in LoVo colon cancer cells via the EP2 and EP4 receptors. (HSU, Hsi-Hsien et al, International journal of molecular sciences,2017, vol.18, no 6, p.1132).

Other studies have shown that inhibition of EP2/EP4 reduces the growth and survival of endometriosis lesions; reducing angiogenesis and innervation of endometriosis lesions; inhibiting the pro-inflammatory state of dorsal root ganglion neurons to reduce pelvic pain; reducing the pro-inflammatory, estrogen dominated, and progesterone resistant molecular milieu of endometrial and endometriosis lesions; and restoring endometrial receptivity by a variety of mechanisms (AROSH, Joe A. et al, Proceedings of the National Academy of Sciences,2015, vol.112, no 31, p.9716-9721) (Banu, S.K.; Lee, J.; Speights, V.O. Jr.; Starzinski-Powitz, A.; Arosh, J.A. molecular Endocrinology 2009,23,1291-1305), and novel selective EP4 antagonists have been identified for the treatment of endometriosisStefan et al, Identification of a bezimidazole acidic Derivative (BAY 1316957) as a Point and selected Human Prostaglandin E2 Receptor Subtype 4(hEP4-R) antioxidant for the Treatment of environmental Chemistry, journal of medical Chemistry, 2019.

EP2 and EP4 in immune responses

In tumors of different entities, all four receptors are related, but expression of EP2 and EP4 receptors predominates on immune cells rather than tissue-resident cells.

PGE 2-induced EP2 receptor signaling also plays an important role in suppressing anti-tumor immune responses. Indeed, most immunomodulatory effects of PGE2 on immune cells are the result of signaling through EP2 and EP4 receptors (KALINSKI, pawel. the Journal of Immunology,2012, vol.188, no1, p.21-28). This is probably due to the fact that signaling through both receptors is transduced by the same G α s stimulating protein and leads to an increase in cAMP intracellular concentration upon activation. This increase in cAMP was shown to be responsible for the inhibition of T Helper (TH)1 cells and the associated decrease in IL-2 and IFN γ, which is important since CD4+ TH cells represent the key effector arm of the adaptive immune system required for cancer control. (O' CALLAGHAN, G.; HOUSTON, A.British journal of pharmacology,2015, vol.172, no 22, p.52).

PGE2 also inhibits the activity of NK cells and cytotoxic T Cells (CTL) in a manner mediated by EP2 and EP4 receptors (HOLT, Dawn et al, Journal of immunotherapy,2012, vol.35, no 2, p.179), both cell types, which may also form part of the anti-tumor immune response. Furthermore, in order to directly suppress the activity of immune cells, the development of Treg cells is promoted by signaling through EP2 and EP4 receptors. Treg cells are potent inhibitors of the immune system and inhibit the activity of a number of immune cells including Dendritic Cells (DCs) (NARENDRA, Bodduluru Lakshmi et al infection Research,2013, vol.62, no 9, p.823-834). DC play a central role in the initiation of tumor-specific immune responses, and the presence of DC in tumors is associated with improved prognosis. Signalling through the EP2 (and EP4) receptors blocks not only the activity of DCs through induction of Treg cells, but also their production by monocytes, leading to the development of immunosuppressive MDSCs from monocytes (DE KEIJZER, Sandra et al, International journal of molecular sciences,2013, vol.14, no 4, p.6542-6555).

Activation of the PGE2/EP2 and PGE2/EP4 signaling pathways positively modulates PD-1 levels in CD8+ T cells infiltrated in lung cancer patients. (WANG, Jinhong et al, Oncology letters,2018, vol.15, no1, p.552-558).

Other studies have shown that PGE2 produced by thyroid cancer cells inhibits the expression of NK activating receptors through EP2 and EP4 receptors on NK cells, inhibiting the cytolytic activity of NK cells. (PARK, Arum, et al. frontiers in immunology,2018, vol.9).

On the other hand, it has been shown that specific EP4 blockade of the EP4 antagonist E704 reverses PGE 2-induced myeloid-mediated immunosuppression, and that E7046 in combination with IL-2-diphtheria toxin fusion protein E7777, which preferentially kills tregs, synergistically alleviates myeloid and Treg-derived immunosuppression within TMEs to promote strong anti-tumor immune responses, providing a combination therapy for cancer patients who may benefit from tumors rich in myeloid and Treg cells (ALBU, Diana I et al, oncoimmulology, 2017, vol.6, no 8, p.e 1338239).

The problem to be solved by the present invention is therefore to provide compounds which are modulators of the EP4 and/or EP2 receptors for prostaglandin E2(PGE 2).

The authors of the present invention have found novel N-benzyl-2-phenoxybenzamide derivatives which may conveniently be substituted as potent modulators of the EP4 and/or EP2 receptors of PGE 2. Thus, the compounds are useful for treating diseases or conditions mediated by modulation of EP4 and/or EP2 receptors, such as cancer, pain such as acute and chronic pain, inflammation such as osteoarthritis, rheumatoid arthritis, neurodegenerative diseases, endometriosis, and kidney disease.

Disclosure of Invention

In a first aspect (aspect 1), the present invention relates to novel N-benzyl-2-phenoxybenzamide derivatives of formula (I) and pharmaceutically acceptable salts thereof:

wherein:

-A represents a group selected from phenyl and a five-or six-membered heteroaryl group having one, two or three heteroatoms selected from N, S and O,

-R¹and R²Independently represent a hydrogen atom, a halogen atom and C_1-3Radical of alkyl, or R¹And R²Together with the carbon atom to which they are attached form C_3-4A cycloalkyl group,

-R³、R⁴、R⁵and each R⁶Independently represent a hydrogen atom,Halogen atom, cyano group, straight or branched C_1-3Haloalkyl, straight-chain or branched C_1-3Alkyl radical, C_3-4Cycloalkyl, straight or branched C_1-3The group of alkoxy and pyridyl,

m is an integer of 1 to 5.

Other aspects of the invention are:

aspect 2) a process for preparing the compound of aspect 1.

Aspect 3) a pharmaceutical composition comprising a therapeutically effective amount of a compound of aspect 1.

Aspect 4) the pharmaceutical composition according to aspect 3, further comprising a therapeutically effective amount of chemotherapeutic agents, anti-inflammatory agents, steroids, immunotherapeutic agents and other agents such as therapeutic antibodies.

Aspect 5) the compound of aspect 1 for use in the treatment of a disease ameliorated by modulation of EP4 and/or EP2 receptors for prostaglandin E2(PGE 2).

The disease ameliorated by modulation of EP4 and/or EP2 receptors may be selected from cancer, pain, inflammation, neurodegenerative disease and kidney disease. The cancer is preferably selected from colon cancer, gastric cancer, prostate cancer, lung cancer, hepatocellular cancer, renal cancer and breast cancer.

Aspect 6) use of a compound of aspect 1 or a pharmaceutical composition of aspect 3 or 4 in the manufacture of a medicament for the treatment of a disease ameliorated by modulation of EP4 and/or EP2 receptors.

Aspect 7) a method of treating or preventing a disease ameliorated by modulation of EP4 and/or EP2 receptors by administering to a subject in need of such treatment a compound of aspect 1 or a pharmaceutical composition of aspect 3 or 4.

Aspect 8) a combination product comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and one or more therapeutic agents selected from chemotherapeutic agents, anti-inflammatory agents, steroids, immunosuppressive agents, therapeutic antibodies, which may be used in combination with a compound of the present application for the treatment of a disease, disorder or condition associated with the modulation of the receptors for prostaglandin E2, EP4 and/or EP 2. One or more additional pharmaceutical agents may be administered to the patient simultaneously or sequentially.

Exemplary chemotherapeutic agents include proteasome inhibitors (e.g., bortezomib), chemotherapeutic agents for treating CNS cancers, including temozolomide, carboplatin, carmustine (BCNU), cisplatin, cyclophosphamide, etoposide, irinotecan, lomustine (CCNU), methotrexate, procarbazine, vincristine, and other chemotherapeutic agents such as thalidomide, ranidomide, and DNA damaging agents such as levo-sarcolysin, doxorubicin, cyclophosphamide, vincristine, etoposide, carmustine, and the like.

Exemplary anti-inflammatory compounds include aspirin, choline salicylate, celecoxib, diclofenac potassium, diclofenac sodium with misoprostol, diflunisal, etodolac, fenoprofen, flurbiprofen, ibuprofen, ketoprofen, meclofenamate sodium, mefenamic acid, nabumetone, naproxen sodium, oxaprozin, piroxicam, rofecoxib, salsalate, sodium salicylate, sulindac, tolmetin sodium, valdecoxib, and the like.

Exemplary steroids include corticosteroids such as cortisone, dexamethasone, hydrocortisone, methylprednisolone, prednisolone, prednisone, and the like.

Exemplary immunosuppressive agents include azathioprine, chlorambucil, cyclophosphamide, cyclosporine, daclizumab, infliximab, methotrexate, tacrolimus, and the like.

Examples of therapeutic antibodies for use in combination therapy include, but are not limited to, trastuzumab (e.g., anti-HER 2), ranibizumab (e.g., anti-VEGF-a), bevacizumab (e.g., anti-VEGF), panitumumab (e.g., anti-EGFR), cetuximab (e.g., anti-EGFR), rituximab (anti-CD 20), and antibodies against c-MET.

In yet another aspect, the present invention relates to a combination product comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and one or more immunotherapeutic agents useful in the treatment of a cancer selected from colon cancer, gastric cancer, prostate cancer, lung cancer and breast cancer.

In a preferred embodiment, the combination product comprises a compound of formula (I) or a pharmaceutically acceptable salt thereof, and one or more immunotherapeutic agents selected from: anti-CTLA 4 antibody selected from ipilimumab and tremelimumab, anti-PD 1 antibodies such as nivolumab, pembrolizumab, cimeprimab (cemipimab), pirlizumab, spartalizumab, MEDI0680, REGN2810 and AMP-224, anti-PDL 1 antibodies such as atelizumab (atezolizumab), avilumab (avelumab), duvacizumab (durvalumab) and MDX-1105, and monoclonal antibodies targeting glycolipid GD2 such as dinotuximab. The components of the combination product are in the same formulation or in separate formulations.

Accordingly, the derivatives of the present invention and pharmaceutically acceptable salts and pharmaceutical compositions comprising such compounds and/or salts thereof are useful in methods of treating pathological conditions or diseases in humans comprising administering to a subject in need of such treatment an effective amount of an N-benzyl-2-phenoxybenzamide derivative of the present invention or a pharmaceutically acceptable salt thereof.

As previously mentioned, the N-benzyl-2-phenoxybenzamide derivatives of the invention are useful in the treatment or prevention of diseases known to be susceptible to amelioration by treatment with EP4 modulators and/or the EP2 receptor of prostaglandin E2. The disease is selected from the group consisting of cancer, such as gastric, colorectal, prostate, lung, hepatocellular, renal and breast cancer, pain, inflammation, neurodegenerative diseases and kidney diseases.

As used herein, the term halogen (halogen) atom includes a chlorine, fluorine, bromine or iodine atom, preferably a fluorine, chlorine or bromine atom, more preferably a fluorine or chlorine atom. The term halo (halo), when used as a prefix, has the same meaning.

As used herein, the term C_1-3Haloalkyl is intended to denote C substituted by one or more halogen atoms, preferably one, two or three halogen atoms_1-3An alkyl group. Preferably, the halogen atom is selected from fluorine or chlorine atoms, more preferably fluorine atoms. In a preferred embodiment, haloalkyl is C substituted with three fluorine atoms (trifluoromethyl)₁An alkyl group.

As used herein, the term C_1-3Alkyl is used to indicate havingStraight or branched chain hydrocarbon group (C) of 1 to 3 carbon atoms_nH_2n+1). Examples include methyl, ethyl, n-propyl, isopropyl radicals.

As used herein, the term C_n-C_mCycloalkyl groups include hydrocarbon monocyclic groups having n to m carbon atoms, for example 3 to 6 or 3 to 4 carbon atoms. Such cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

As used herein, the term C_1-3For containing a straight or branched chain C bound to an oxygen atom_1-3Alkyl radical (C)_nH_2n+1-O-). The preferred alkoxy group is methoxy.

As used herein, the term C_3-4Cycloalkoxy is taken to mean a compound containing C attached to an oxygen atom_3-4Radical of cycloalkyl (radial).

As used herein, the term five to six membered heteroaryl is used to denote a heteroaromatic ring containing carbon, hydrogen and one, two or three heteroatoms, preferably selected from N, O and S, as part of the ring, such as furan, pyridine, pyrimidine, pyrazine, pyrrole, imidazole, pyrazole, oxazole, thiazole and thiophene, preferably furan, pyridine, pyrazine, pyrrole, imidazole, pyrazole, oxazole, thiazole and thiophene. Said radicals may optionally be substituted by one or more substituents as defined in each case. Preferred groups are optionally substituted pyridyl, pyrimidinyl. When the heteroaryl group bears two or more substituents, these substituents may be the same or different.

As used herein, some of the atoms, groups, chains or rings present in the general structures of the present invention are "optionally substituted. This means that these atoms, groups, chains or rings may be unsubstituted or substituted in any position by one or more, for example 1,2,3 or 4, substituents, whereby a hydrogen atom is bonded to an unsubstituted atom, group, chain or ring by a chemically acceptable atom, radical, chain or ring. When two or more substituents are present, each substituent may be the same or different.

As used herein, the term pharmaceutically acceptable salt is used to denote a salt in combination with a pharmaceutically acceptable base. Pharmaceutically acceptable bases include alkali metal (e.g., sodium or potassium), alkaline earth metal (e.g., calcium or magnesium) hydroxides, and organic bases such as alkylamines, aralkylamines, and heterocyclic amines.

The term "modulator" refers to a molecule that blocks or otherwise interferes with a particular biological activity of a receptor.

As used herein, the term "Ki" refers to the concentration that causes half-maximal inhibition of control specific binding. Ki value can be derived from IC₅₀The concentration of radioligand in the assay and the affinity of the radioligand for the receptor. IC (integrated circuit)₅₀The values can be estimated from the appropriate dose response curve, more precisely, the IC₅₀Values may be determined using non-linear regression analysis.

According to one embodiment of the invention, in the compounds of formula (I), R¹And R²Independently selected from hydrogen atom and C_1-3Alkyl, or R¹And R²Together with the carbon atom to which they are attached form C_3-4A cycloalkyl group; r³Selected from hydrogen atoms and halogen atoms; r⁴And R⁵Represents a hydrogen atom.

According to one embodiment of the invention, in the compounds of formula (I), R¹、R²、R³、R⁴And R⁵Represents a hydrogen atom.

According to one embodiment of the invention, in the compounds of formula (I), a is selected from phenyl, pyridyl, pyrazolyl, pyrimidinyl, thiophenyl and furanyl, preferably from phenyl, pyridyl, pyrazolyl, pyrimidinyl and thiophenyl; more preferably from phenyl, pyridyl, pyrimidinyl and thiophenyl; even more preferably from phenyl, pyridyl and thiophenyl.

According to one embodiment of the invention, a represents phenyl.

In a preferred embodiment, each R is⁶Independently represents a group selected from:

a) a hydrogen atom, and a nitrogen atom,

b) halogen atom

c) The cyano group(s),

d) carbamoyl (-CONH)₂) A base, and

d) straight or branched C_1-3A haloalkyl group.

In a specific embodiment, each R⁶Independently represents a group selected from:

a) a hydrogen atom, and a nitrogen atom,

b) a fluorine atom,

c) a chlorine atom in the presence of a chlorine atom,

c) the cyano group(s),

d) carbamoyl (-CONH)₂) A base, and

d) a trifluoromethyl group.

In a more preferred embodiment, A represents phenyl and each R represents⁶Independently represents a group selected from fluorine, trifluoromethyl and cyano, and m is an integer of 1 to 4.

According to another embodiment of the invention, a represents a five or six membered heteroaromatic ring having one or two N atoms as part of the ring. In a preferred embodiment, A represents a group selected from pyridyl or thiophenyl, each R represents⁶Independently represents a group selected from fluorine, trifluoromethyl and cyano, and m is an integer of 1 to 4.

According to another embodiment of the invention, R¹、R²、R³、R⁴And R⁵Represents a hydrogen atom, A represents a phenyl group and each R⁶Independently represents a group selected from fluorine or trifluoromethyl, and m is an integer of 1 to 5.

According to another embodiment, m is an integer from 1 to 4, preferably m is an integer from 1 to 2.

According to another embodiment, A represents a group selected from phenyl, pyridyl, pyrimidyl, phenylthio, R¹、R²、R³、R⁴、R⁵Is a hydrogen atom, each R⁶Independently represents a group selected from a hydrogen atom, fluorine, chlorine, cyano, trifluoromethyl and pyridyl, and m is an integer of 1 to 2.

Specific individual compounds of the invention include:

4- ((2- (4-fluorophenoxy) -5- (pyridin-4-yl) benzamido) methyl) benzoic acid

4- ((4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((4 '-chloro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((4 '-fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((3 '-fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((2 '-fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((4 '-cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((3 '-cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (3-fluoropyridin-4-yl) benzamido) methyl) benzoic acid)

4- ((2- (4-fluorophenoxy) -5- (pyridin-3-yl) benzamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (3-chloropyridin-4-yl) benzamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (pyrimidin-5-yl) benzamido) methyl) benzoic acid

4- ((3',4' -difluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((4- (4-fluorophenoxy) -4'- (trifluoromethyl) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (pyridin-2-yl) benzamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (pyrimidin-4-yl) benzamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (phenylthio-2-yl) benzamido) methyl) benzoic acid

4- ((3',5' -difluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl)) benzoic acid

4- ((3 '-chloro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((3' -fluoro-4- (4-fluorophenoxy) -5' - (trifluoromethyl) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((3' -carbamoyl-5 ' -chloro-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-ylcarboxamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (1H-pyrazol-4-yl) benzamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (1-methyl-1H-pyrazol-4-yl) benzamido) methyl) benzoic acid

4- ((2- (4-fluorophenoxy) -5- (1-methyl-1H-pyrazol-5-yl) benzamido) methyl) benzoic acid

4- ((4- (4-fluorophenoxy) -3'- (pyridin-4-yl) - [1,1' -biphenyl ] -3-ylcarboxamido) methyl) benzoic acid

4- ((5- (5-carbamoylfuran-2-yl) -2- (4-fluorophenoxy) benzamido) methyl) benzoic acid

3-fluoro-4- ((4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl ] -3-ylcarboxamido) methyl) benzoic acid

(R) -4- (1- (4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl ] -3-carboxamido) ethyl) benzoic acid

4- (1- (4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl ] -3-carboxamido) cyclopropyl) benzoic acid

4- ((3' -fluoro-5 ' -cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid

4- ((3' -chloro-5 ' -cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl ] -3-carboxamido) methyl) benzoic acid.

The compounds of the present invention can be prepared by using the following methods. For ease of describing the process, specific examples are used, but they do not limit the scope of the invention in any way.

Scheme 1

Reagents and conditions: (a) MeOH, H₂SO₄At 70 ℃; b) NaH, DMF, 120 ℃, overnight; c) NaOH, MeOH/H₂O, room temperature for 3 hours; d) HATU, DIPEA, DCM, room temperature, overnight; e) [1,1' -bis (diphenylphosphino) ferrocene]Dichloropalladium (II), Cs₂CO₃dioxane/H₂O, 100 ℃ overnight, f) NaOH, MeOH/H₂O, room temperature, overnight.

The compounds of formula (I) may be prepared by the synthetic route described in scheme 1.

Starting with a 5-bromo-2-fluorobenzoic acid derivative of formula (II), reaction with methanol to form the ester (III), and reaction with a phenol derivative of formula (IV) gives the compound of formula (V).

Hydrolysis of the compound of formula (V) under basic aqueous conditions gives an intermediate of formula (VI) which is reacted with an amine of formula (VII) in the presence of a coupling agent such as HATU, a lead compound of formula (VIII). Aryl bromides of formula (VIII) may be converted to compounds of formula (I) by reaction with boronic acids or esters under palladium-catalyzed conditions followed by hydrolysis under basic aqueous conditions.

Scheme 2

Reagents and conditions: g) [1,1' -bis (diphenylphosphino) ferrocene]Dichloropalladium (II), Cs₂CO₃dioxane/H₂O, 100 ℃, overnight, h) NaH, DMF, 120 ℃, overnight; i) NaOH, MeOH/H₂O, room temperature and overnight; ) HATU, DIPEA, DCM, room temperature, overnight; k) NaOH, MeOH/H₂O, room temperature, overnight.

An alternative route to the compounds of formula (I) is depicted in scheme 2. Reacting an aryl bromide of formula (III) with a boronic acid, boronic ester, zinc reagent or tributylstannane reagent in the presence of a palladium catalyst to produce an intermediate of formula (X). Using reactions similar to those described in scheme 1, after hydrolysis of a compound of formula (X) with phenol (IV) under basic aqueous conditions, an intermediate of formula (XI) is formed, which is reacted with an amine of formula (VII) in the presence of a coupling agent such as HATU to provide the corresponding compound of formula (I) after hydrolysis under basic aqueous conditions.

The synthesis of the compounds of the present invention is illustrated by the following examples, including the preparation of intermediates, but not in any way limiting the scope of the invention.

Abbreviations

The following abbreviations are used in the present application, with corresponding definitions:

RT: at room temperature

HATU: n- [ (dimethylamino) -1H-1,2, 3-triazolo- [4,5-b ] pyridin-1-ylmethylene ] -N-methylmethanemethanemethanamine hexafluorophosphate N-oxide

DIPEA: n, N-diisopropylethylamine

DMF: dimethyl formamide

DCM: methylene dichloride

THF: tetrahydrofuran (THF)

DMSO, DMSO: dimethyl sulfoxide

Pharmacological Activity

Results

The compounds of the invention were tested for binding to the human EP1, EP2, EP3 and EP4 receptors, and the IC50 assay was tested at several concentrations.

Competitive binding in the human EP1 receptor:

prostaglandin EP1 receptor competition binding assay with binding buffer (Mes 10mM, EDTA1mM, MgCl)₂1mM, pH 6.0) in a polypropylene 96-well microplate. Mu.g of membranes from the HEK-EP1 cell line and prepared in our laboratory (batch: A003/19-10-2011, protein concentration 1756. mu.g/ml), 3nM [3H ] were incubated in each well]Prostaglandin E2(169.8Ci/mmol, 0.1mCi/ml, Perkin Elmer NET428250UC) and the compounds studied and the standards. Nonspecific binding was determined in the presence of PGE 210. mu.M (Cayman 14010). The reaction mixture (Vt: 250. mu.l/well) was incubated at 25 ℃ for 60 minutes, and after filtration, 200. mu.L was transferred to GF/C96-well plates (Millip)ore, Madrid, Spain) and four washes with 250. mu.l wash buffer (Mes 10mM, EDTA1mM, MgCl₂1mM, pH 6.0) and then measured in a microplate beta scintillation counter (Microbeta Trilux, PerkinElmer, Madrid, Spain). (Abramovitz et al, Biochim Biophys Acta 2000; 1483: 285-293).

Competitive binding in the human EP2 receptor:

prostaglandin EP2 receptor competition binding assay with binding buffer (Hepes 50mM, CaCl)₂ 1mM，MgCl₂5mM, pH 7.4) in a polypropylene 96-well microplate. Mu.g of membranes from the EP2 cell line (Millipore HTS185M, protein concentration 2000. mu.g/ml), 7.5nM [3H ] were incubated in each well]Prostaglandin E2(169.8Ci/mmol, 0.1mCi/ml, Perkin Elmer NET428250UC) and the compounds studied and standards. Nonspecific binding was determined in the presence of PGE 2200. mu.M (Cayman 14010). The reaction mixture (Vt: 250. mu.l/well) was incubated at 25 ℃ for 90 minutes, after filtration 200. mu.L was transferred to GF/C96-well plates (Millipore, Madrid, Spain) and washed four times with 250. mu.l of wash buffer (Hepes 50mM, NaCl 500mM, pH 7.4) and then measured in a microplate beta scintillation counter (Microbeta Trilux, Perkinelmer, Madrid, Spain). (Wilson et al, Br J Pharmacol 2006; 148: 326-.

Competitive binding in the human EP3 receptor:

prostaglandin EP3 receptor competition binding experiments using binding buffer (Tris-HCl 50mM, MgCl)₂10mM, EDTA1mM, pH 7.4) pretreated multi-screen GF/C96 well plates (Millipore, Madrid, Spain). Mu.g of membranes from the EP3 cell line (Millipore; HTS092M, protein concentration 2000. mu.g/ml), 1.5nM [3H ] were incubated in each well]Prostaglandin E2(169.8Ci/mmol, 0.1mCi/ml, Perkin Elmer NET428250UC) and the compounds studied and standards. Nonspecific binding was determined in the presence of PGE 2200. mu.M (Cayman 14010). The reaction mixture (Vt: 250. mu.l/well) was incubated at 25 ℃ for 90 minutes, after filtration 200. mu.l were transferred to GF/C96-well plates (Millipore, Madrid, Spain) and washed four times with 250. mu.l of wash buffer (Tris-HCl 50mM, pH 7.4) before plating in a microplate beta scintillation counter (Microbeta Trilux, Perkine)lmer, Madrid, Spain). (Audoly et al, Mol Pharmacol 1997; 51: 61-68).

Competitive binding in the human EP4 receptor:

prostaglandin EP4 receptor competition binding assays were performed using binding buffer (Mes 25mM, EDTA1mM, MgCl)₂10mM, pH 6.0) pretreated multi-screen GF/B96 well plates (Millipore, Madrid, Spain). Mu.g of membranes from the HEK-EP4 cell line and prepared in our laboratory (batch: A003/27-04-2011; protein concentration 2803. mu.g/ml), 1nM [3H ] were incubated in each well]Prostaglandin E2(169.8Ci/mmol, 0.1mCi/ml, Perkin Elmer NET428250UC) and the compounds studied and standards. Nonspecific binding was determined in the presence of PGE 210. mu.M (Cayman 14010). The reaction mixture (Vt: 250. mu.l/well) was incubated at 25 ℃ for 120 minutes, after filtration 200. mu.l were transferred to GF/B96-well plates (Millipore, Madrid, Spain) and washed six times with 250. mu.l of wash buffer (Mes 25mM, BSA 0.01%, pH 6.0) and then measured in a microplate beta scintillation counter (Microbeta Trilux, Perkinelmer, Madrid, Spain). (Abramovitz et al, Biochim Biophys Acta 2000; 1483: 285-293).

Inhibition constants (Ki) were calculated using the Cheng Prusoff equation

Where L ═ the concentration of radioligand in the assay, and KD ═ the affinity of the radioligand for the receptor. Scatchard plots were used to determine KD.

The compounds of the present invention do not show any significant binding affinity to the receptors EP1 and EP 3.

Table 1 shows Ki values for receptors EP2 and EP4 for some compounds of the invention.

Ki range: a < 0.1. mu.M; 0.1 μ Μ < ═ B <1 μ Μ; 1 μ M < ═ C <10 μ M, D > -10 μ M

TABLE 1

As can be seen from the results described in table 1, the compounds of the present invention effectively bind to EP4 and/or EP2 receptors of prostaglandin E2.

The compounds of the invention are useful for the treatment or prevention of diseases known to be susceptible to amelioration by modulation of the EP4 and/or EP2 receptors. Such diseases are selected from the group consisting of cancer, pain, inflammation, neurodegenerative diseases and kidney diseases.

The compounds of the invention and their salts are therefore useful for the preparation of medicaments for the treatment or prevention of diseases which are known to be susceptible to amelioration by modulation of the EP4 and/or EP2 receptors.

Accordingly, the derivatives of the present invention and pharmaceutically acceptable salts thereof and pharmaceutical compositions comprising such compounds and/or salts thereof are useful in methods of treating or preventing human conditions known to be susceptible to amelioration by modulation of the EP4 and/or EP2 receptors, which methods comprise administering to a subject in need of such treatment or prevention an effective amount of an N-benzyl-2-phenoxybenzamide derivative of the present invention or a pharmaceutically acceptable salt thereof.

An "effective amount" or "therapeutically effective amount" of a compound, drug or pharmacologically active agent refers to an amount of the compound, drug or agent that is non-toxic but sufficient to provide the desired effect. The amount of "effective" will vary from subject to subject, depending on the age and general condition of the subject, the particular active agent or agents, and the like. Therefore, it is not always possible to specify an exact "effective amount". However, one of ordinary skill in the art can use routine experimentation to determine an appropriate "effective" amount in any individual case.

One therapeutic use of the compounds of the present invention is in the treatment of proliferative diseases or disorders, such as cancer. The cancer is selected from colon cancer, gastric cancer, prostate cancer, lung cancer, hepatocellular carcinoma, renal cancer, and breast cancer.

The present invention also provides a pharmaceutical composition comprising as active ingredient at least a N-benzyl-2-phenoxybenzamide derivative of formula (I) or a pharmaceutically acceptable salt thereof in association with other therapeutic agents and pharmaceutically acceptable excipients such as carriers or diluents. Depending on the nature of the formulation and whether further dilution is required prior to administration, the active ingredient may be present at from 0.001% to 99%, preferably from 0.01% to 90%, by weight of the composition. Preferably, the composition is prepared in a form suitable for oral, topical, nasal, rectal, transdermal or injectable administration.

Pharmaceutically acceptable excipients which are mixed with the active compound or a salt of such a compound to form the compositions of the invention are well known per se, and the actual excipient used depends, inter alia, on the intended method of administering the composition.

The compositions of the present invention are preferably suitable for injection and oral administration. In this case, the composition for oral administration may take the form of tablets, delayed (retard) tablets, sublingual tablets, capsules, inhalation aerosols, inhalation solutions, dry powder inhalants or liquid preparations, such as mixtures, elixirs, syrups or suspensions, all containing the compound of the present invention; such formulations may be prepared by methods well known in the art.

Diluents which may be used in the preparation of the compositions include those liquid and solid diluents which are compatible with the active ingredient, as well as colouring or flavouring agents, if desired. Tablets or capsules may conveniently contain from 2 to 500mg of the active ingredient or an equivalent amount of the salt thereof.

Liquid compositions suitable for oral use may be in the form of solutions or suspensions. The solution may be an aqueous solution of a soluble salt or other derivative of the active compound combined with, for example, sucrose to form a syrup. Suspensions may contain the insoluble active compounds of the invention, or a pharmaceutically acceptable salt thereof, in association with water, and a suspending or flavoring agent.

Parenteral injection compositions can be prepared from soluble salts, which may or may not be lyophilized, and can be dissolved in pyrogen-free aqueous media or other suitable parenteral injection solutions.

The effective dose is generally in the range of 2-2000mg of active ingredient per day. The daily dose may be administered in one or more treatments per day, preferably from 1 to 4 treatments.

The invention will be further illustrated by the following examples. The following is given by way of illustration and in no way limits the scope of the invention.

Examples

General procedure (General.) reagents, solvents and starting products were obtained from commercial sources. The term "concentration" means evaporation in vacuo using a Buchi rotary evaporator. When indicated, the reaction product was purified by "flash" chromatography on silica gel (40-63 μm) with the indicated solvent system. The spectral data were measured on a Varian Mercury 400 spectrometer. HPLC-MS was measured in a Buchi 535 instrument. HPLC-MS was performed on a Gilson instrument equipped with a Gilson 321 piston pump, a Gilson 864 vacuum degasser, a Gilson 189 sample module, 1/1000Gilson shunt, Gilson 307 pump, Gilson 170 detector, and Thermoquest Fennigan aQa detector.

Intermediate 1: 4- (Azidomethyl) -3-fluorobenzoic acid methyl ester

A solution of methyl 4- (bromomethyl) -3-fluorobenzoate (100mg, 1.0eq) and sodium azide (29mg, 1.2eq) in DMF (1.0mL) was stirred at 80 ℃ for 90 min. Once cooled, the reaction was diluted with brine (3mL) and extracted with ethyl acetate (3X 10 mL). The combined organic solutions were washed with water (25mL) and brine (25mL) and washed with Na₂SO₄Dried, filtered and concentrated under reduced pressure. The desired azide was obtained as a pale yellow liquid (quantitative) which was used in the next step without further purification.

¹H-NMR(400MHz，DMSO-d⁶)：δ＝7.82(dd，1H)，7.74(dd，1H)，7.64(t，1H)，4.62(s，2H)，3.87(s，3H)。

HPLC-MS：Rt 2.65 m/z 182.0(MH⁺)

Intermediate 2: 4- (aminomethyl) -3-fluorobenzoic acid methyl ester

A solution of methyl 4- (azidomethyl) -3-fluorobenzoate (110mg, 1.0eq), triphenylphosphine (189mg, 1.2eq) and water (0.9mL) in THF (2mL) was stirred at room temperature for 20 h. The solvent was removed under reduced pressure and purified by flash column chromatography (dichloromethane: methanol, 30: 1) to give methyl 4- (aminomethyl) -3-fluorobenzoate (70mg, yield 63.7%) as a yellow wax.

¹H-NMR(400MHz，DMSO-d⁶)：δ＝7.78(dd，1H)，7.67(t，1H)，7.61(dd，1H)，3.85(s，3H)，3.80(s，2H)，1.96(s，2H)。

HPLC-MS：Rt 1.63 m/z 184.1(MH⁺)

Intermediate-3: 4-fluoro- [1,1' -biphenyl]-3-Carboxylic acid methyl ester

To 4-fluoro- [1,1' -biphenyl]-3-Carboxylic acid (200.0mg, 0.93mmol) in methanol (1mL) with the addition of a catalytic amount of H₂SO₄And the reaction was stirred at 70 ℃ overnight. The solution was partitioned between water and ethyl acetate and saturated NaHCO₃And the organic layer was washed with brine. Dried over anhydrous sodium sulfate, filtered and concentrated to give intermediate 1(167.0mg, 76.0%) as an oil.

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.09(dd，1H)，7.96(ddd，1H)，7.68(d，2H)，7.46(m，2H)，3.89(s，3H)。

HPLC-MS：Rt 5.337 m/z 231.0(MH⁺)

The following intermediates were synthesized using the procedure described for intermediate 3:

intermediate 4: 5-bromo-2-fluorobenzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝7.98(dd，1H)，7.87(m，1H)，7.37(dd，1H)，3.86(s，3H)。

Intermediate 5: 5-bromo-2- (4-fluorophenoxy) benzoic acid methyl esterEsters

To a stirred solution of 4-fluorophenol (481.0mg, 4.29mmol) and sodium cyanide (171.6mg, 4.29mmol) in N, N-dimethylformamide was added a solution of methyl 5-bromo-2-fluorobenzoate (1000mg, 4.29mmol) in N, N-dimethylformamide. The solution was heated at 120 ℃ and stirred for 16 hours. After cooling to room temperature, the mixture was diluted with water (20mL) and extracted with ethyl acetate (3 × 20 mL). The organic layer was dried over sodium sulfate, filtered and concentrated under reduced pressure to give the title compound as a pale yellow oil (1028.3mg, 73.7%).

¹H-NMR(400MHz，DMSO-d⁶)：δ＝7.95(d，1H)，7.75(dd，1H)，7.22(m，2H)，7.04(m，2H)，6.95(d，1H)，3.75(s，3H)。

HPLC-MS：Rt 5.602 m/z 326.9(MH⁺)

The following intermediates were synthesized using the procedure described for intermediate 5 and the corresponding methyl ester and 4-fluorophenol:

intermediate 6: 4- (4-fluorophenoxy) - [1,1' -biphenyl]-3-Carboxylic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.07(d，1H)，7.88(dd，1H)，7.68(d，2H)，7.49(t，2H)，7.39(t，1H)，7.23(t，2H)，7.10(s，1H)，7.06(m，2H)，3.77(s，3H)。

HPLC-MS：Rt 5.948 m/z 323.0(MH⁺)

Intermediate 7: 5-bromo-2- (4-fluorophenoxy) benzoic acid

To a solution of methyl 5-bromo-2- (4-fluorophenoxy) benzoate (1101.3mg, 3.39mmol) in methanol (8mL) and THF (8mL) was added NaOH 2M (8.5mL, 16.94mmol), and the mixture was stirred at room temperature for 3 hours. The solution was diluted with water (15mL) and the pH adjusted to 4.0 by addition of HCl 1M. The mixture was extracted with ethyl acetate (3 × 50mL), and the combined organic layers were dried over sodium sulfate and concentrated, washed with pentane to give a white solid (823.5mg, 78.1%).

¹H-NMR(400MHz，DMSO-d⁶)：δ＝13.24(s，1H)，7.92(s，1H)，7.72(d，1H)，7.21(t，2H)，7.01(s，2H)，6.94(d，1H)。

HPLC-MS：Rt 3.087 m/z 312.9(MH⁺)

The following intermediates were synthesized using the procedure described for intermediate 7 and the corresponding methyl esters:

intermediate 8: 4- (4-fluorophenoxy) - [1,1' -biphenyl]-3-carboxylic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝13.04(s，1H)，8.06(d，1H)，7.84(dd，1H)，7.68(d，2H)，7.48(t，2H)，7.39(t，1H)，7.22(t，2H)，7.04(m，3H)。

HPLC-MS：Rt 3.435 m/z 309.0(MH⁺)

Intermediate 9: 4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl]-3-carboxylic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝13.09(s，1H)，8.13(d，1H)，7.95(m，4H)，7.72(m，2H)，7.23(m，2H)，7.05(m，2H)。

HPLC-MS：Rt2.20 m/z 377.0(MH⁺)

Intermediate 10: 4- ((5-bromo-2- (4-fluorophenoxy) benzamido) methyl) benzoic acid methyl ester

To a solution of 5-bromo-2- (4-fluorophenoxy) benzoic acid (600mg, 1.93mmol) in anhydrous DCM (15mL) was added HATU (880.0mg, 2.30mmol) and DIPEA (1.31mL, 7.71 mmol). At room temperature under N₂After stirring the reaction for 15 minutes, methyl 4- (aminomethyl) benzoate hydrochloride (466.7mg, 2.30mmol) was added and the reaction was left in the chamberStir at room temperature overnight. After adding 20ml of H₂After O, the aqueous phase was extracted with DCM 3X 20 mL. The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to give a white solid (728.5mg, 82.4%).

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.97(t，1H)，7.85(d，2H)，7.79(d，1H)，7.63(dd，1H)，7.38(d，2H)，7.24(t，2H)，7.11(dd，2H)，6.86(d，1H)，4.51(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 5.523 m/z 459.0(MH⁺)

The following intermediates were synthesized using the procedure described for intermediate 10 and the corresponding chemical reagents or derivatives:

intermediate 11: (R) -4- (1- (5-bromo-2- (4-fluorophenoxy) benzamido) ethyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.89(d，1H)，7.84(d，2H)，7.69(d，1H)，7.62(dd，1H)，7.45(d，2H)，7.22(t，2H)，7.07(m，2H)，6.89(d，1H)，5.08(p，1H)，3.83(s，3H)，1.38(d，3H)。

HPLC-MS：Rt 3.21 m/z 473.9(MH⁺)

Intermediate 12: 4- (1- (5-bromo-2- (4-fluorophenoxy) benzamido) cyclopropyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.13(s，1H)，7.78(s，1H)，7.76(d，2H)，7.65(dd，1H)，7.24(m，4H)，7.08(m，2H)，6.95(d，1H)，3.82(s，3H)，1.28(m，2H)，1.21(m，2H)。

HPLC-MS：Rt3.18 m/z 485.9(MH⁺)

Intermediate 13: 4- ((4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.96(t，1H)，7.93(d，1H)，7.86(d，2H)，7.76(dd，1H)，7.68(d，2H)，7.48(t，2H)，7.39(m，3H)，7.25(t，2H)，7.13(m，2H)，7.00(d，1H)，4.54(d，2H)，3.84(s，3H)。

HPLC-MS：Rt 5.843 m/z 456.1(MH⁺)

Intermediate 14: 4- ((5-bromo-2- (4-fluorophenoxy) benzamido) methyl) benzoic acid

To a solution of methyl 4- ((5-bromo-2- (4-fluorophenoxy) benzamido) methyl) benzoate (200mg, 1.0eq) in THF (2.0mL) and methanol (2.0mL) was added NaOH (2M) (1.1mL, 5.0eq)), and the mixture was stirred at room temperature for 20 hours. After dilution with water (10mL), the mixture was neutralized with aqueous HCl, filtered, and washed with water and pentane to obtain the acid as a white solid (176mg, 90.9% yield).

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，8.98(t，1H)，7.82(m，3H)，7.63(dd，1H)，7.32(m，2H)，7.24(m，2H)，7.12(m，2H)，6.86(d，1H)，4.49(d，2H)。

HPLC-MS：Rt 1.26 m/z 444.0(MH⁺)

Intermediate 15: 4- ((2- (4-fluorophenoxy) -5- (pyridin-4-yl) benzamido) methyl) benzoic acid methyl ester

To a solution of methyl 4- ((5-bromo-2- (4-fluorophenoxy) benzamido) methyl) benzoate (70.0mg, 0.15mmol) in dioxane, pyridine-4-boronic acid (37.6mg, 0.31mmol), [1,1' -bis (diphenylphosphino) ferrocene](II) solution of Palladium (II) dichloride (7.5mg, 0.009mmol) with Cs added₂CO₃2M (0.23mL, 0.46 mmol). Mixing the mixture with N₂Degassed and stirred at 100 ℃ overnight. Then, the reaction was filtered through celite and extracted with ethyl acetate. The organic portion was treated with NaOH 1M, saturated NaHCO₃And brine, and dried over anhydrous sodium sulfate. The solvent was removed under vacuum and the residue was purified using flash chromatography (hexane: ethyl acetate 1: 1). The solvent was removed and washed with pentane/diethyl ether to give the product as a white solid (44.7mg, 65.3%).

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.00(t，1H)，8.65(d，2H)，8.08(d，1H)，7.89(m，3H)，7.75(d，2H)，7.42(d，2H)，7.27(m，2H)，7.17(m，2H)，7.01(d，1H)，4.56(d，2H)，3.84(s，3H)。

HPLC-MS：Rt 4.875 m/z 457.1(MH⁺)

The following intermediates were synthesized using the procedure described for intermediate 15 using the corresponding chemical reagents or derivatives:

intermediate 16: 4- ((4 '-chloro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.99(t，1H)，7.92(d，1H)，7.85(d，2H)，7.74(m，3H)，7.51(t，2H)，7.39(d，2H)，7.25(t，2H)，7.12(m，2H)，6.99(d，1H)，4.53(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 6.203 m/z 490.1(MH⁺)

Intermediate 17: 4- ((4 '-fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.98(s，1H)，7.86(m，3H)，7.72(s，3H)，7.37(m，2H)，7.27(m，4H)，7.12(s，2H)，6.98(d，1H)，4.53(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 5.945 m/z 474.1(MH⁺)

Intermediate 18: 4- ((3 '-fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.99(s，1H)，7.95(d，1H)，7.85(d，2H)，7.80(dd，1H)，7.53(t，3H)，7.40(d，2H)，7.24(dd，3H)，7.13(dd，2H)，6.98(d，1H)，4.54(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 5.963 m/z 474.1(MH⁺)

Intermediate 19: 4- ((2 '-fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.99(s，1H)，7.85(d，3H)，7.64(d，1H)，7.57(s，1H)，7.40(d，3H)，7.30(m，4H)，7.17(s，2H)，6.98(d，1H)，4.54(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 5.936 m/z 474.1(MH⁺)

Intermediate 20: 4- ((4 '-cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.01(t，1H)，8.02(d，1H)，7.93(m，4H)，7.85(m，3H)，7.41(d，2H)，7.27(t，2H)，7.16(m，2H)，7.00(d，1H)，4.55(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 5.649 m/z 481.1(MH⁺)

Intermediate 21: 4- ((3 '-cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.00(t，1H)，8.20(s，1H)，8.04(dd，2H)，7.85(t，4H)，7.68(t，1H)，7.41(d，2H)，7.27(t，2H)，7.15(dd，2H)，7.00(d，1H)，4.55(d，2H)，3.84(s，3H)。

HPLC-MS：Rt 5.668 m/z 481.1(MH⁺)

Intermediate 22: 4- ((2- (4-fluorophenoxy) -5- (3-fluoropyridin-4-yl) benzamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.03(t，1H)，8.67(d，1H)，8.51(d,1H)，7.97(s，1H)，7.86(d，2H)，7.77(d，1H)，7.68(dd，1H)，7.42(d，2H)，7.29(dd，2H)，7.20(m，2H)，7.01(d，1H)，4.56(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 5.199 m/z 475.1(MH⁺)

Intermediate 23: 4- ((2- (4-fluorophenoxy) -5- (pyridin-3-yl) benzamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.00(t,1H)，8.92(d，1H)，8.58(dd，1H)，8.11(m，1H)，7.99(d，1H)，7.84(m，3H)，7.50(dd，1H)，7.41(d，2H)，7.26(t，2H)，7.14(m，2H)，7.02(d，1H)，4.54(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 5.015 m/z 457.1(MH⁺)

Intermediate 24: 4- ((2- (4-fluorophenoxy) -5- (3-chloropyridin-4-yl) benzamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.02(t，1H)，8.75(s，1H)，8.59(d，1H)，7.86(d，2H)，7.84(d，1H)，7.64(dd，1H)，7.53(d，1H)，7.42(d，2H)，7.30(t，2H)，7.21(m，2H)，6.98(d，1H)，4.55(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 5.378 m/z 491.1(MH⁺)

Intermediate 25: 4- ((2- (4-fluorophenoxy) -5- (pyrimidin-5-yl) benzamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.18(m，3H)，9.02(t，1H)，8.08(d，1H)，7.88(m，3H)，7.42(d，2H)，7.27(t，2H)，7.15(m，2H)，7.04(d，1H)，4.55(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 4.715 m/z 458.1(MH⁺)

Intermediate 26: 4- ((3',4' -difluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.98(t，1H)，7.95(d，1H)，7.81(m，4H)，7.53(m，2H)，7.40(d，2H)，7.26(t，2H)，7.13(m，2H)，6.98(d，1H)，4.54(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 5.977 m/z 492.1(MH⁺)

Intermediate 27: 4- ((4- (4-fluorophenoxy) -4'- (trifluoromethyl) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.02(t，1H)，8.01(d，1H)，7.93(d，2H)，7.84(m，5H)，7.41(d，2H)，7.27(dd，2H)，7.16(m，2H)，7.01(d，1H)，4.55(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 6.223 m/z 524.1(MH⁺)

Intermediate 28: 4- ((4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.00(t，1H)，8.01(m，3H)，7.85(d，3H)，7.71(m，2H)，7.40(d，2H)，7.26(t，2H)，7.15(dd，2H)，7.01(d，1H)，4.55(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 6.199 m/z 524.1(MH⁺)

Intermediate 29: 4- ((2- (4-fluorophenoxy) -5- (phenylthio-2-yl) benzamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.99(t，1H)，7.90(d，1H)，7.85(d，2H)，7.74(dd，1H)，7.56(d，1H)，7.52(d，1H)，7.39(d，2H)，7.25(t，2H)，7.13(m，3H)，6.95(d，1H)，4.52(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 3.21 m/z 462.1(MH+)

Intermediate 30: 4- ((3',5' -difluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.00(t，1H)，8.00(d，1H)，7.84(m，3H)，7.48(m，2H)，7.41(d，2H)，7.25(m，3H)，7.14(m，2H)，6.97(d，1H)，4.54(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 3.31 m/z 492.21(MH⁺)

Intermediate 31: 4- ((3 '-chloro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.99(t，1H)，7.96(d，1H)，7.86(d，2H)，7.80(dd,1H)，7.76(m，1H)，7.66(d，1H)，7.50(t，1H)，7.43(m，3H)，7.25(m，2H)，7.14(m，2H)，6.87(d，1H)，4.54(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 3.41 m/z 490.16(MH⁺)

Intermediate 32: 4- ((3' -fluoro-4- (4-fluorophenoxy) -5' - (trifluoromethyl) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.01(t，1H)，8.06(d，1H)，7.95(d，1H)，7.88(m，4H)，7.67(d，1H)，7.41(d，2H)，7.26(m，2H)，7.15(m，2H)，7.00(d，1H)，4.55(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 3.48 m/z 542.21(MH⁺)

Intermediate 33: 4- ((3' -cyano-5 ' -fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.00(t，1H)，8.12(s，1H)，8.07(d，1H)，8.00(m，1H)，7.88(m，4H)，7.42(d，2H)，7.26(m，2H)，7.15(m，2H)，6.99(d，1H)，4.53(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 3.13 m/z 499.21(MH⁺)

Intermediate 34: 4- ((3' -chloro-5 ' -cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.00(t，1H)，8.22(d，1H)，8.16(t，1H)，8.05(m，2H)，7.87(m，3H)，7.42(d，2H)，7.27(m，2H)，7.15(m，2H)，6.98(d，1H)，4.55(d，2H)，3.84(s，3H)。

HPLC-MS：Rt 3.26 m/z 515.1(MH⁺)

Intermediate 35: 4- ((2- (4-fluorophenoxy) -5- (1H-pyrazol-4-yl) benzamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.83(s，1H)，8.99(t，1H)，7.82(m，4H)，7.62(m，2H)，7.36(dd，2H)，7.24(m，2H)，7.12(m，2H)，6.86(d，1H)，4.50(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 2.03 m/z 446.0(MH⁺)

Intermediate 36: 4- ((2- (4-fluorophenoxy) -5- (1-methyl-1H-pyrazol-4-yl) benzamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.92(t，1H)，8.18(s，1H)，7.84(m，4H)，7.64(dd，1H)，7.37(d，2H)，7.21(m，2H)，7.06(m，2H)，6.93(d，1H)，4.51(d，2H)，3.86(s，3H)，3.83(s，3H)。

HPLC-MS：Rt 2.70 m/z 460.2(MH⁺)

Intermediate 37: 4- ((2- (4-fluorophenoxy) -5- (1-methyl-1H-pyrazol-5-yl) benzamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.01(t，1H)，7.85(d，2H)，7.77(d，1H)，7.61(dd，1H)，7.48(d，1H)，7.40(d，2H)，7.26(m，2H)，7.17(m，2H)，6.98(d，1H)，6.43(d，1H)，4.54(d，2H)，3.86(s，3H)，3.83(s，3H)。

HPLC-MS：Rt 2.73 m/z 460.13(MH⁺)

Intermediate body 38: 4- ((4- (4-fluorophenoxy) -3'- (pyridin-4-yl) - [1,1' -biphenyl)]-3-ylcarboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.00(t，1H)，8.66(dd，2H)，8.06(m，2H)，7.85(m，7H)，7.63(t，1H)，7.40(d，2H)，7.25(m，2H)，7.14(m，2H)，7.02(d，1H)，4.55(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 3.06 m/z 533.2(MH⁺)

Intermediate 39: 4- ((5- (5-Cyanofuran-2-yl) -2- (4-fluorophenoxy) benzamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.02(t，1H)，8.10(s，1H)，7.87(m，3H)，7.42(m，3H)，7.28(m，4H)，7.18(m，2H)，6.97(d，1H)，4.55(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 3.08 m/z 471.1(MH⁺)

Intermediate 40: 3-fluoro-4- ((4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl]-3-ylcarboxamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.01(t，1H)，8.01(m，2H)，7.86(dd，1H)，7.74(dd，2H)，7.67(m，2H)，7.44(t，1H)，7.25(m，2H)，7.15(m，2H)，7.01(d，1H)，4.55(d，2H)，3.85(s，3H)。

Intermediate 41: (R) -4- (1- (4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl)]-3-carboxamido) ethyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝8.89(d，1H)，8.01(m，2H)，7.91(d，1H)，7.84(m，3H)，7.72(m，2H)，7.48(d，2H)，7.24(t，2H)，7.11(m，2H)，7.03(d，1H)，5.14(p，1H)，3.83(s，3H)，1.41(d，3H)。

HPLC-MS：Rt3.51 m/z 538.0(MH⁺)

Intermediate body 42: 4- (1- (4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl)]-3-carboxamido) cyclopropyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.13(s，1H)，8.04(d，2H)，7.96(d，1H)，7.86(dd，1H)，7.75(m，4H)，7.27(m，4H)，7.13(dd，2H)，7.09(d，1H)，3.82(s，3H)，1.32(t，2H)，1.25(t，2H)。

HPLC-MS：Rt 3.48 m/z 550.0(MH⁺)

Intermediate 43: 4- ((2- (4-fluorophenoxy) -5- (pyridin-2-yl) benzamido) methyl) benzoic acid methyl ester

To a solution of methyl 4- ((5-bromo-2- (4-fluorophenoxy) benzamido) methyl) benzoate (100.0mg, 0.22mmol) and tetrakis (triphenylphosphine) palladium (0) (15.1mg, 0.013mmol) in dry dioxane (1.5mL) was added 2- (tributylstannyl) pyridine (113.4mg, 0.26 mmol). Mixing the mixture with N₂Purged and stirred at room temperature for 2 days. The reaction was diluted with ethyl acetate and washed with NaOH 2M. Drying the organic layer over anhydrous sodium sulfate, filteringAnd concentrated and the residue purified by flash chromatography (hexane: ethyl acetate 1: 1) to give a white solid. The product was washed with pentane/diethyl ether (40.3mg, 40.5%).

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.00(t，1H)，8.66(d，1H)，8.39(d，1H)，8.16(dd，1H)，7.98(d，1H)，7.90(dd，1H)，7.86(d，2H)，7.42(d，2H)，7.36(dd，1H)，7.27(t，2H)，7.17(dd，2H)，6.98(d，1H)，4.56(d，2H)，3.84(s，3H)。

HPLC-MS：Rt 5.308 m/z 457.1(MH⁺)

The following intermediates were synthesized using the procedure described for intermediate 43 and the corresponding chemical reagents or derivatives:

intermediate 44: 4- ((2- (4-fluorophenoxy) -5- (pyrimidin-4-yl) benzamido) methyl) benzoic acid methyl ester

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.24(d，1H)，9.04(t，1H)，8.86(d，1H)，8.51(d，1H)，8.29(dd，1H)，8.12(dd，1H)，7.87(d，2H)，7.44(d，2H)，7.30(m，2H)，7.22(m，2H)，7.00(d，1H)，4.57(d，2H)，3.83(s，3H)。

HPLC-MS：Rt 4.890 m/z 458.1(MH⁺)

Example 1: 4- ((2- (4-fluorophenoxy) -5- (pyridin-4-yl) benzamido) methyl) benzoic acid

To a solution of methyl 4- ((2- (4-fluorophenoxy) -5- (pyridin-4-yl) benzamido) methyl) benzoate (35.0mg, 0.077mmol) in methanol (0.5mL) and THF (0.5mL) was added NaOH 2M (0.19mL, 0.38mmol), and the mixture was stirred at room temperature overnight. The solution was diluted with water (15mL) and the pH adjusted to 4.0 by addition of HCl 1M, and the precipitate was filtered and washed with pentane (27.8mg, 82.0%).

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.85(s，1H)，9.01(t，1H)，8.64(d，2H)，8.07(d，1H)，7.90(dd，1H)，7.84(d，2H)，7.74(d，2H)，7.39(d，2H)，7.27(t，2H)，7.17(dd，2H)，7.01(d，1H)，4.55(d，2H)。

HPLC-MS：Rt 3.362 m/z 443.1(MH⁺)

The following intermediates were synthesized using the procedure described in example 1 and the corresponding chemicals or derivatives:

example 2: 4- ((4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.84(s，1H)，8.95(s，1H)，7.93(d，1H)，7.83(d，2H)，7.76(dd，1H)，7.68(d，2H)，7.48(t，2H)，7.38(t，3H)，7.25(t，2H)，7.13(m，2H)，7.00(d，1H)，4.53(d，2H)。

HPLC-MS：Rt 3.867 m/z 442.1(MH⁺)

Example 3: 4- ((4 '-chloro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，8.97(t，1H)，7.93(d，1H)，7.83(d，2H)，7.75(m，3H)，7.53(d，2H)，7.37(d，2H)，7.26(t，2H)，7.13(m，2H)，6.99(d，1H)，4.53(d，2H)。

HPLC-MS：Rt 4.221 m/z 476.0(MH⁺)

Example 4: 4- ((4 '-fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.87(s，1H)，8.97(t，1H)，7.90(d，1H)，7.83(d，2H)，7.73(m，3H)，7.36(d，2H)，7.29(m，4H)，7.12(m，2H)，6.99(d，1H)，4.53(d，2H)。

HPLC-MS：Rt 4.029 m/z 460.1(MH⁺)

Example 5: 4- ((3 '-fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，8.98(t，1H)，7.96(d，1H)，7.81(m，3H)，7.53(m，3H)，7.37(d，2H)，7.22(m，3H)，7.14(m，2H)，6.99(d，1H)，4.53(d，2H)。

HPLC-MS：Rt 4.033 m/z 460.1(MH⁺)

Example 6: 4- ((2 '-fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.87(s，1H)，8.98(s，1H)，7.83(s，3H)，7.64(d，1H)，7.57(s，1H)，7.34(m，7H)，7.18(s，2H)，6.99(d，1H)，4.53(d，2H)。

HPLC-MS：Rt 4.001 m/z 460.1(MH⁺)

Example 7: 4- ((4 '-cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，9.00(t，1H)，8.02(s，1H)，7.91(m，4H)，7.84(d，3H)，7.38(d，2H)，7.27(t，2H)，7.16(m，2H)，7.00(d，1H)，4.54(d，2H)。

HPLC-MS：Rt 3.864 m/z 467.1(MH⁺)

Example 8: 4- ((3 '-cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，8.99(t，1H)，8.20(s，1H)，8.04(dd，2H)，7.84(d，4H)，7.68(t，1H)，7.38(d，2H)，7.27(t，2H)，7.15(dd，2H)，7.00(d，1H)，4.54(d，2H)。

HPLC-MS：Rt 3.860 m/z 467.1(MH⁺)

Example 9: 4- ((2- (4-fluorophenoxy) -5- (3-fluoropyridin-4-yl) benzamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，9.01(t,1H)，8.67(d，1H)，8.51(d，1H)，7.97(s，1H)，7.84(d，2H)，7.77(d，1H)，7.68(dd，1H)，7.39(d，2H)，7.29(t，2H)，7.20(m，2H)，7.01(d，1H)，4.55(d，2H)。

HPLC-MS：Rt 3.495 m/z 461.1(MH⁺)

Example 10: 4- ((2- (4-fluorophenoxy) -5- (pyridin-3-yl) benzamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，8.99(s，1H)，8.92(d，1H)，8.58(dd，1H)，8.11(m，1H)，7.99(d，1H)，7.83(m，3H)，7.50(dd，1H)，7.38(d，2H)，7.26(t，2H)，7.15(m，2H)，7.02(d，1H)，4.54(d，2H)。

HPLC-MS：Rt 3.390 m/z 443.1(MH⁺)

Example 11: 4- ((2- (4-fluorophenoxy) -5- (3-chloropyridin-4-yl) benzamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，9.01(t，1H)，8.75(s，1H)，8.59(d，1H)，7.83(dd，3H)，7.64(dd，1H)，7.53(d，1H)，7.39(d，2H)，7.30(t，2H)，7.21(dd，2H)，6.98(d，1H)，4.54(d，2H)。

HPLC-MS：Rt 3.624 m/z 477.0(MH⁺)

Example 12: 4- ((2- (4-fluorophenoxy) -5- (pyrimidin-5-yl) benzamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，9.17(m，3H)，9.01(t，1H)，8.08(d，1H)，7.88(m，3H)，7.42(d，2H)，7.27(t，2H)，7.15(m，2H)，7.04(d，1H)，4.54(d，2H)。

HPLC-MS：Rt 3.147 m/z 444.1(MH⁺)

Example 13: 4- ((3',4' -difluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.89(s，1H)，8.97(t，1H)，7.95(d，1H)，7.79(m，4H)，7.54(m，2H)，7.37(d，2H)，7.26(t，2H)，7.13(dd，2H)，6.98(d，1H)，4.53(d，2H)。

HPLC-MS：Rt 4.084 m/z 478.1(MH⁺)

Example 14: 4- ((4- (4-fluorophenoxy) -4'- (trifluoromethyl) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，9.00(t，1H)，8.01(d，1H)，7.93(d，2H)，7.83(dd，5H)，7.38(d，2H)，7.27(t，2H)，7.16(m，2H)，7.02(d，1H)，4.54(d，2H)。

HPLC-MS：Rt 2.53 m/z 510.57(MH⁺)

Example 15: 4- ((4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝δ＝12.88(s，1H)，9.01(t，1H)，8.01(dd，3H)，7.84(dd，3H)，7.72(q，2H)，7.37(d，2H)，7.60(m，2H)，7.15(m，2H)，7.01(d，1H)，4.54(d，2H)。

HPLC-MS：Rt 2.47 m/z 510.51(MH⁺)

Example 16: 4- ((2- (4-fluorophenoxy) -5- (pyridin-2-yl) benzamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，8.99(t，1H)，8.67(d，1H)，8.39(d，1H)，8.16(dd，1H)，7.99(d，1H)，7.89(m，1H)，7.84(d，2H)，7.39(d，2H)，7.35(m，1H)，7.27(t，2H)，7.17(dd，2H)，6.99(d，1H)，4.55(d，2H)。

HPLC-MS：Rt 3.502 m/z 433.1(MH⁺)

Example 17: 4- ((2- (4-fluorophenoxy) -5- (pyrimidin-4-yl) benzamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，9.24(d，1H)，9.03(t，1H)，8.86(d，1H)，8.51(d，1H)，8.28(dd，1H)，8.12(dd，1H)，7.85(d，2H)，7.41(d，2H)，7.30(m，2H)，7.22(dd，2H)，7.00(d，1H)，4.56(d，2H)。

HPLC-MS：Rt 3.243 m/z 444.1(MH⁺)

Example 18: 4- ((2- (4-fluorophenoxy) -5- (phenylthio-2-yl) benzamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，8.98(t，1H)，7.90(d，1H)，7.82(dd，2H)，7.73(dd，1H)，7.56(d，1H)，7.53(dd，1H)，7.35(，2H)，7.24(t，2H)，7.12(m，3H)，6.96(d，1H)，4.52(d，2H)。

HPLC-MS：Rt 2.18 m/z 448.1(MH⁺)

Example 19: 4- ((3',5' -difluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid.

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.87(s，1H)，8.99(t，1H)，8.00(d，1H)，7.83dd，3H)，7.49(m，2H)，7.38(d，2H)，7.25(m，3H)，7.14(m，2H)，6.98(d，1H)，4.54(d，2H)。

HPLC-MS：Rt 2.30 m/z 478.17(MH⁺)

Example 20: 4- ((3 '-chloro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid.

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，8.99(t，1H)，7.97(d，1H)，7.80(m，4H)，7.68(d，1H)，7.51(t，1H)，7.46(m，1H)，7.39(d，2H)，7.27(m，2H)，7.15(m，2H)，7.00(d，1H)，4.55(d，2H)。

HPLC-MS：Rt 2.86 m/z 476.13(MH⁺)

Example 21: 4- ((3' -fluoro-4- (4-fluorophenoxy) -5' - (trifluoromethyl) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid.

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.89(s，1H)，9.01(t，1H)，8.08(d，1H)，7.96(d，1H)，7.90(m，2H)，7.84(d，2H)，7.70(d，1H)，7.39(d，2H)，7.28(m，2H)，7.17(m，2H)，7.01(d，1H)，4.56(d，2H)。

HPLC-MS：Rt 2.46 m/z 528.19(MH⁺)

Example 22: 4- ((3' -carbamoyl-5)'-chloro-4- (4-fluorophenoxy) - [1,1' -biphenyl]-3-ylcarboxamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.89(s，1H)，9.00(t，1H)，8.26(s，1H)，8.15(s，1H)，8.05(d，1H)，7.94(m，1H)，7.88(m，1H)，7.84(m，2H)，7.63(s，1H)，7.38(d，2H)，7.26(m，2H)，7.15(m，2H)，7.01(d，1H)，4.54(d，2H)。

HPLC-MS：Rt 1.96 m/z 519.07(MH⁺)

Example 23: 4- ((2- (4-fluorophenoxy) -5- (1H-pyrazol-4-yl) benzamido) methyl) benzoic acid.

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.96(s，1H)，8.90(t，1H)，8.09(s，2H)，7.85(d，1H)，7.81(m，2H)，7.69(dd，1H)，7.33(m，2H)，7.22(m，2H)，7.06(m，2H)，6.94(d，1H)，4.49(d，2H)。

HPLC-MS：Rt 1.06 m/z 432.1(MH⁺)

Example 24: 4- ((2- (4-fluorophenoxy) -5- (1-methyl-1H-pyrazol-4-yl) benzamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，8.91(t，1H)，8.18(s，1H)，7.88(s，1H)，7.81(m，3H)，7.64(dd，1H)，7.33(d，2H)，7.22(m，2H)，7.06(m，2H)，6.94(d，1H)，4.50(d，2H)，3.86(s，3H)。

HPLC-MS：Rt 1.78 m/z 446.10(MH⁺)

Example 25: 4- ((2- (4-fluorophenoxy) -5- (1-methyl-1H-pyrazol-5-yl) benzamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，9.00(t，1H)，7.83(d，2H)，7.77(d，1H)，7.61(dd，1H)，7.48(d，1H)，7.37(d，2H)，7.27(m，2H)，7.17(m，2H)，6.98(d，1H)，6.43(t，1H)，4.53(d，2H)，3.86(s，3H)。

HPLC-MS：Rt 1.82 m/z 446.10(MH⁺)

Example 26：4-((4- (4-fluorophenoxy) -3'- (pyridin-4-yl) - [1,1' -biphenyl]-3-ylcarboxamido) methyl) benzoic acid.

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.88(s，1H)，9.02(t，1H)，8.92(d，1H)，8.38(d，2H)，8.25(s，1H)，8.10(d，1H)，7.98(d，1H)，7.92(d，2H)，7.83(d，2H)，7.71(t，1H)，7.38(d，2H)，7.27(m，2H)，7.15(m，2H)，7.04(d，1H)，4.54(d，2H)。

HPLC-MS：Rt 2.15 m/z 519.2(MH⁺)

Example 27: 4- ((5- (5-carbamoylfuran-2-yl) -2- (4-fluorophenoxy) benzamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.89(s，1H)，9.00(t，1H)，8.17(t，1H)，8.02(s，1H)，7.94(m，2H)，7.45(s，2H)，7.37(t，2H)，7.26(m，2H)，7.13(m，3H)，6.99(d，1H)，4.53(d，2H)。

HPLC-MS：Rt 1.74 m/z 475.0(MH⁺)

Example 28: 3-fluoro-4- ((4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl]-3-ylcarboxamido) methyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝13.26(s，1H)，9.00(t，1H)，8.00(m，3H)，7.86(dd，1H)，7.73(m，2H)，7.63(m，2H)，7.41(t，1H)，7.26(m，2H)，7.15(m，2H)，7.00(d，1H)，4.54(d，2H)。

HPLC-MS：Rt 1.54 m/z 528.0(MH⁺)

Example 29: (R) -4- (1- (4- (4-fluorophenoxy) -3'- (trifluoromethyl) - [1,1' -biphenyl)]-3-carboxamido) ethyl) benzoic acid¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.86(s，1H)，8.88(d，1H)，8.06(m，2H)，7.90(t，1H)，7.84(dd，3H)，7.72(q，2H)，7.45(d，2H)，7.25(t，2H)，7.12(m，2H)，7.03(d，1H)，5.13(p，1H)，1.40(t，3H)。

HPLC-MS：Rt 2.45 m/z 524.0(MH⁺)

Example 30: 4- (1- (4- (4-fluorophenoxy)3'- (trifluoromethyl) - [1,1' -biphenyl]-3-carboxamido) cyclopropyl) benzoic acid

¹H-NMR(400MHz，DMSO-d⁶)：δ＝9.09(s，1H)，8.04(d，2H)，7.95(s，1H)，7.86(d，1H)，7.73(d，4H)，7.27(t，2H)，7.18(d，2H)，7.13(dd，2H)，7.08(d，1H)，1.27(t，2H)，1.22(t，2H)。

HPLC-MS：Rt 2.79 m/z 536.0(MH⁺)

Example 31: 4- ((3' -fluoro-5 ' -cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid

To a solution of 4- ((5-bromo-2- (4-fluorophenoxy) benzamido) methyl) benzoic acid (80mg, 1.0eq) and (3-cyano-5-fluorophenyl) boronic acid (59mg, 2.0eq) in DMF (1.2mL) was added 2M Cs2CO3(0.27mL, 3.0eq) and pd (dppf) Cl2 · DCM (9mg, 0.06eq) under a nitrogen atmosphere. After stirring at 110 ℃ for 20 h, the mixture was filtered through celite, washing with ethyl acetate (20 mL). The organic phase was washed with saturated aqueous sodium bicarbonate (20mL) and brine (20mL) over Na₂SO₄Dried, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (hexane: ethyl acetate, 1: 1) to obtain acid IV-484-536(22mg, yield 25.2%) as a white solid.

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.86(s，1H)，8.99(t，1H)，8.12(s，1H)，8.07(d，2H)，8.01(dd，1H)，7.89(dd，1H)，7.84(d，2H)，7.39(d，2H)，7.27(m，2H)，7.16(m，2H)，6.99(d，1H)，4.54(d，2H)。

HPLC-MS：Rt 2.23 m/z 485.1(MH⁺)

Example 32: 4- ((3' -chloro-5 ' -cyano-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-carboxamido) methyl) benzoic acid

To 4- ((3' -cyano-5 ' -fluoro-4- (4-fluorophenoxy) - [1,1' -biphenyl)]-3-ylcarboxamido) methyl) benzoic acid methyl ester (100mg, 1.0eq) in THF (1.0mL) HCl (4M) (3.0mL) was added and the mixture was stirred at 80 ℃ for 46 h. After dilution with water (5mL), the mixture was extracted with ethyl acetate (3X 10mL) and Na₂SO₄Dried, filtered and concentrated under reduced pressure. The crude material was purified by flash column chromatography (hexane: ethyl acetate, 1: 1) to obtain the acid as a white solid (18mg, yield 18.6%).

¹H-NMR(400MHz，DMSO-d⁶)：δ＝12.87(s，1H)，8.98(t，1H)，8.22(s，1H)，8.16(t，1H)，8.07(d，1H)，8.03(m，1H)，7.89(dd，1H)，7.84(d，2H)，7.39(d，2H)，7.27(m，2H)，7.16(m，2H)，6.99(d，1H)，4.54(d，2H)。

HPLC-MS：Rt 2.34 m/z 501.1(MH⁺)