RECEPTOR 1 AND LPA RECEPTOR 2 TNHTBTTORS

FIELD OF THE INVENTION

The present invention generally relates to compounds inhibiting lysophosphatidic acid receptors (hereinafter LPA inhibitors); the invention relates to compounds that are 5-membered heterocyclyl derivatives, methods of preparing such compounds, pharmaceutical compositions containing them and therapeutic use thereof.

The compounds of the invention may be useful for instance in the treatment of many disorders associated with LPA receptors mechanisms.

BACKGROUND OF THE INVENTION

Lysophosphatidic acid (LPA) is a phospholipid mediator concentrated in serum that acts as a potent extracellular signaling molecule through at least six cognate G protein- coupled receptors (GPCRs) in numerous developmental and adult processes including cell survival, proliferation, migration, differentiation, vascular regulation, and cytokine release.

These LPA-mediated processes involve nervous system function, vascular development, immune system function, cancer, reproduction, fibrosis, and obesity (see e.g. Yung etal, J Lipid Res. 2014 Jul;55(7): 1192-214). The formation of an LPA species depends on its precursor phospholipid, which can vary typically by acyl chain length and degree of saturation. The term LPA generally refers to 18:1 oleoyl-LPA (1 -acyl-2 - hydroxy-sn-glycero3 -phosphate), that is the most quantitatively abundant forms of LPA in human plasma with 16:0-, 18:2-, and 18: 1-LPA (see e.g. Sano et ah, J Biol Chem. 2002 Dec 13; 277(50):21197 206). All LPA species are produced from membrane phospholipids via two major metabolic routes. Depending upon the site of synthesis, membrane phospholipids get converted to the corresponding lysophospholipids by the action of phospholipase A1 (PLA1), phospholipase A2 (PLA2), or PLA1 and lecithin- cholesterol acyltransferase (LCAT). Autotaxin (ATX) then acts on the lysophospholipids and converts them into LPA species. The second pathway first converts the phospholipids into phosphatidic acid by the action of phospholipase D. Then PLA1 or PLA2 metabolize phosphatidic acid to the lysophosphatidic acids (see e.g. Riaz et al, Int J Mol Sci. 2016 Feb; 17(2): 215).

ATX activity is the maj or source of plasma extracellular LPA but the source of tissue LPA that contributes to signaling pools likely involves not only ATX but other enzymes as well. The biological functions of LPA are mediated by at least six recognized cell- surface receptors.

All LPA receptors are rhodopsin-like 7-TM proteins that signal through at least two of the four Ga subunit families (Gal2/13, Gaq/11, Gai/o and GaS). LPA receptors usually trigger response from multiple heterotrimeric G-proteins, resulting in diverse outcomes in a context and cell type dependent manner. Gal 2/ 13 -mediated LPA signaling regulates cell migration, invasion and cytoskeletal re-adjustments through activation of RHO pathway proteins. RAC activation downstream of Gai/o-PI3K also regulates similar processes, but the most notable function of LPA-induced Gai/o is mitogenic signaling through the R.AF- MEK-MAPK cascade and survival signaling through the PI3K-AKT pathway. The LPA- coupled Gaq/11 protein primarily regulates Ca2+ homeostasis through PLC and the second messengers IP3 and DAG. Lastly, GaS can activate adenylyl cyclase and increase cAMP concentration upon LPA stimulation (see e.g. Riaz et ah, Int J Mol Sci. 2016 Feb; 17(2): 215).

LPA, especially LPA1, LPA2 and LPA3, have been implicated in migration, invasion, metastasis, proliferation and survival and differ in their tissue distribution and downstream signaling pathways. LPA1 is a 41-kD protein that is widely expressed, albeit at different levels, in all human adult tissues examined and the importance of LPA1 signaling during development and adult life has been demonstrated through numerous approaches (see e.g. Ye at al, 2002, Neuroreport. Dec 3; 13(17) :2169-75). Wide expression of LPA1 is observed in adult mice, with clear presence in at least brain, uterus, testis, lung, small intestine, heart, stomach, kidney, spleen, thymus, placenta, and skeletal muscle. LPA1 is also widely expressed in humans where the expression is more spatially restricted during embryonic development. LPA1 couples with and activates three types of G proteins: God/o, Gaq/11, and Gal2/13. LPA1 activation induces a range of cellular responses: cell proliferation and survival, cell migration, cytoskeletal changes, Ca2+ mobilization, adenylyl cyclase inhibition and activation of mitogen-activated protein kinase, phospholipase C, Akt, and Rho pathways (see e.g. Choi et al, Annu Rev Pharmacol Toxicol. 2010; 50:157-86).

LPA2 in humans is a 39-kD protein and shares -55% amino acid sequence homology with LPA1 (see e.g. Yung et al, J Lipid Res. 2014 Jul;55(7): 1192-214). In mouse, LPA2 is highly expressed in kidney, uterus, and testis and moderately expressed in lung; in human tissues, high expression of LPA2 is detected in testis and leukocytes, with moderate expression found in prostate, spleen, thymus, and pancreas.

In terms of signaling activity, LPA2 mostly activates the same pathways as triggered by LPA1 with some exceptions that regards its unique cross-talk behavior. For example, LPA2 promotes cell migration through interactions with focal adhesion molecule TRIP6 (see e.g. Lai YJ, 2005, Mol.Cell.Biol. 25:5859 68), and several PDZ proteins and zinc finger proteins are also reported to interact directly with the carboxyl-terminal tail of LPA2 (see e.g. Lin FT, 2008, Biochim.Biophys.Acta 1781:558 62).

Human LPA3 is a 40-kD protein and shares sequence homology with LPA1 (-54%) and LPA2 (-49%). In adult humans LPA3 is highly expressed in heart, pancreas, prostate and testis. Moderate levels of expression are also found in brain, lungs and ovary. Like LPA1 and LPA2 the signaling activity of LPA3 results from its coupling to God/o and Gaq/11 (see e.g Ishii et al, Mol Pharmacol 58:895 902, 2000). Each LPA has multiple important regulatory functions throughout the body.

As LPA signaling has been strongly implicated in many disease states, great interest has been expressed in developing specific LPA inhibitors (see e.g. Stoddard et el, Biomol Ther (Seoul) 2015 Jan;23(l):l-ll). Different studies have demonstrated a positive role for LPA in the pathogenesis of pulmonary fibrosis (PF), a devastating disease characterized by alveolar epithelial cell injury, accumulation of myofibroblasts and deposition of extracellular matrix proteins leading to a loss of lung function and death (see e.g. Wilson MS, Wynn TA (2009), Mucosal Immunol 2: 103 121).

Evidences showed that lysophosphatidic acid levels dramatically increase in bronchoalveolar lavage fluid of PF patients where it mediates fibroblast migration in the injured lung acting through LPA1 (see e.g. Tager etal, Nat Med. 2008 Jan; 14(l):45-54). In addition, mice lacking LPA1 or LPA2 are markedly protected from fibrosis and mortality in a mouse model of the bleomycin induced pulmonary fibrosis (see e.g. Huang et al, Am J Re spir Cell Mol Biol 2013 Dec; 49(6): 912 922 and Tager et al, Nat Med. 2008 Jan; 14(1) :45-54).

In vitro, LPA1 is known to induce the proliferation and differentiation of lung fibroblasts (see e.g. Shiomi etal, Wound Repair Regen. 2011 Mar Apr; 19(2): 229 240), and to augment the fibroblast-mediated contraction of released collagen gels (see e.g. Mio etal, Journal of Laboratory and Clinical Medicine, Volume 139, Issue 1, January 2002, Pages 20-27). In human lung fibroblasts, the knockdown of LPA2 attenuated the LPA- induced expression of TGF-bI and the differentiation of lung fibroblasts to myofibroblasts, resulting in the decreased expression of different profibrotic markers such as FN, a-SMA, and collagen, as well as decreased activation of extracellular regulated kinase 1/2, Akt, Smad3, and p38 mitogen-activated protein kinase (see e.g. Huang et al, Am J Respir Cell Mol Biol. 2013 Dec; 49(6): 912 922). Moreover Xu et al., confirmed that the expression of LPA2 was also up-regulated in lungs from bleomycin-challenged mice where it is able to induce the activation of TGF-b pathway, a key cytokine that play an essential role during the development of the disease, via a RhoA and Rho kinase pathway (see e.g. Xu et al., Am J Pathol. 2009 Apr; 174(4): 1264-79). In in vivo preclinical model, the oral administration of an LPA1 antagonist significantly reduced bleomycin- induced pulmonary fibrosis in mice ( Tager etal, Nat Med. 2008 Jan; 14(1) :45-54; Swaney et al, Br J Pharmacol. 2010 Aug; 160(7): 1699 1713), and the intraperitoneal injection of an LPA1/3 antagonist ameliorated irradiation-induced lung fibrosis (see e.g. Gan et al, 2011, Biochem Biophys Res Commun 409: 7 13). In a renal fibrosis model, LPA1 administration of an LPA1 antagonist suppressed renal interstitial fibrosis (see e.gPradere et al, J Am Soc Nephrol 2007; 18:3110 3118).

Various compounds have been described in the literature as LPA1 or LPA2 antagonist. WO2019126086 and WO2019126087 (Bristol-Myers Squibb) disclose cyclohexyl acid isoxazole azines as LPA1 antagonist, useful for the treatment of disorder or condition associated with dysregulation of lysophosphatidic acid receptor 1.

WO2019126099 (Bristol-Myers Squibb) discloses isoxazole N-linked carbamoyl cyclohexyl acid as LPA1 antagonist for the treatment of disorder or condition associated with dysregulation of lysophosphatidic acid receptor 1.

W02019126090 (Bristol-Myers Squibb) discloses triazole N-linked carbamoyl cyclohexyl acids as LPA1 antagonists. The compounds are selective LPA1 receptor inhibitors and are useful for the treatment of disorder or condition associated with dysregulation of lysophosphatidic acid receptor 1. WO2017223016 (Bristol-Myers Squibb) discloses carbamoyloxymethyl triazole cyclohexyl acids as LPA1 antagonist for the treatment of fibrosis including idiopathic pulmonary fibrosis.

WO2012028243 (Merck) discloses pyrazolopyridinone derivatives according to formula (I) and a process of manufacturing thereof as LPA2 receptor antagonists for the treatment of various diseases.

Amgen Inc. discloses in “Discovery of potent LPA2 (EDG4) antagonists as potential anticancer agents” Bioorg Med Chem Lett. 2008 Feb 1; 18(3): 1037-41, LPA2 antagonists. Key compounds were evaluated in vitro for inhibition of LPA2 mediated Erk activation and proliferation of HCT-116 cells. These compounds could be used as tool compounds to evaluate the anticancer effects of blocking LPA2 signaling.

Of note, antagonizing the LPA receptors may be useful for the treatment of fibrosis and disease, disorder and conditions that result from fibrosis, and even more antagonizing both receptors LPA1 and LPA2 may be particularly efficacious in the treatment of the above-mentioned disease, disorder and conditions.

Despite the above cited prior art, there remains a potential for developing dual inhibitors of both receptors LPA1 and LPA2 useful for the treatment of diseases or conditions associated with a dysregulation of LPA receptors, in particular fibrosis.

In this respect, the state of the art does not describe or suggest 5-membered heterocyclyl derivatives of general formula (I) of the present invention having a dual antagonist activity on both receptors LPA1 and LPA2 which represent a solution to the aforementioned need.

SUMMARY OF THE INVENTION

In a first aspect the invention refers to a compound of formula (I) wherein Xi, X ₂ and X ₃ are independently selected from S, N, C, CH, S(0) ₂ , S(O) provided that at least one is S, S(O) or S(0) ₂ and R ₇ is present only when X ₂ and/or X ₃ are C or N; X _4, X ₅ , Xe and X ₇ are independently N, C or CH wherein no more than two are N;

Ri is absent when X4 is N, or is H or selected from the group consisting of (Ci-C4)alkyl, halo, CN, and -OR2;

Li is a bond or an (Ci-C4)alkylene group;

L2 1S selected from the group consisting of wherein Li and L3 indicate the attachment of L2 to these groups;

L ₃ is a bond or an (Cl-C6)alkylene group wherein said alkylene may be optionally substituted by one or more group selected from (Ci-C4)alkyl, -OR ₃ , -NR ₄ R ₅ , -(Ci-C4)alkylene-OR ₃ , -(Ci-C4)alkylene-NR4R5, halo and oxo; Q is H or selected from the group consisting of (C3-C8)cycloalkyl, (C3- C ₈ )heterocycloalkyl, aryl and 5-6 membered heteroaryl, wherein each of said cycloalkyl, heterocycloalkyl, aryl and heteroaryl may be optionally substituted by one or more group selected from (Ci-C4)alkyl, -NR4R5, -(Ci-C4)alkylene-OR3, -(Ci-C4)alkylene-NR4R5, halo, -NR ₂ C(0)R ₆ , -NR ₂ C(0)0R6, -(Ci-C ₄ )alkylene-NR ₂ C(0)R ₆ and

-(Ci-C ₄ )alkylene-NR ₂ C(0)0R ₆ ;

R2 is H or an (Ci-C ₄ )alkyl group;

R3 is H or selected from the group consisting of (Ci-C ₄ )alkyl, (Ci-C ₄ )haloalkyl, (C ₃ - C8)cycloalkyl and (C ₃ -C ₈ )heterocycloalkyl;

R4 and R5 are at each occurrence independently H or selected from the group consisting of (Ci-C4)alkyl, (C3-C8)cycloalkyl, (Ci-Ce) haloalkyl, (Ci-Ce) hydroxyalkyl, (Ci-Ce) aminoalkyl, (Ci-Ce) alkoxyl, (Ci-Ce) alkoxy-(Ci-C6) alkyl, (C3-C6) heterocycloalkyl- (C1-C6) alkyl, aryl, heteroaryl and (C3-C6) heterocycloalkyl; or

R4 and R5 may form together with the nitrogen atom to which they are attached a 5 or 6 membered saturated heterocyclic ring system optionally containing a further heteroatom selected from N, S or O, said heterocyclic ring system may be optionally substituted by one or more groups selected from (Ci-C4)alkyl, -CN, (Ci-Ce) haloalkyl, (Ci-Ce) hydroxyalkyl,

(Ci-Ce) aminoalkyl, halogen and oxo;

Re is H or selected from the group consisting of (Ci-C ₄ )alkyl, (Ci-Ce) haloalkyl, (Ci-Ce) hydroxyalkyl, (Ci-Ce) aminoalkyl and (Ci-Ce) alkoxyl,

R7 is H or selected from (Ci-C ₄ )alkyl and halo.

In a second aspect, the invention refers to pharmaceutical composition comprising a compound of formula (I) or a stereoisomer, tautomer or pharmaceutically acceptable salt or solvate thereof, in admixture with one or more pharmaceutically acceptable carrier or excipient. In a third aspect, the invention refers to a compound of formula (I) for use as a medicament.

In a further aspect, the invention refers to a compound of formula (I) or a stereoisomer, tautomer or pharmaceutically acceptable salt or solvate thereof for use in treating disease, disorder, or condition associated with dysregulation of lysophosphatidic acid receptor 1 (LPA1) and 2 (LPA2).

In a further aspect, the invention refers to a compound of formula (I) or a stereoisomer, tautomer or pharmaceutically acceptable salt or solvate thereof for use in the prevention and/or treatment of fibrosis and/or diseases, disorders, or conditions that involve fibrosis.

In a further aspect, the invention refers to a compound of formula (I) or a stereoisomer, tautomer or pharmaceutically acceptable salt or solvate thereof for use in the prevention and/or treatment idiopathic pulmonary fibrosis (IPF).

DETAILED DESCRIPTION OF THE INVENTION

The term “pharmaceutically acceptable salts”, as used herein, refers to derivatives of compounds of formula (I) wherein the parent compound is suitably modified by converting any of the free acid or basic group, if present, into the corresponding addition salt with any base or acid conventionally intended as being pharmaceutically acceptable.

Suitable examples of said salts may thus include mineral or organic acid addition salts of basic residues such as amino groups, as well as mineral or organic basic addition salts of acid residues such as carboxylic groups.

Cations of inorganic bases which can be suitably used to prepare salts comprise ions of alkali or alkaline earth metals such as potassium, sodium, calcium or magnesium.

Those obtained by reacting the main compound, functioning as a base, with an inorganic or organic acid to form a salt comprise, for example, salts of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, methane sulfonic acid, camphor sulfonic acid, acetic acid, oxalic acid, maleic acid, fumaric acid, succinic acid and citric acid.

The term "solvate" means a physical association of a compound of this invention with one or more solvent molecules, whether organic or inorganic. This physical association includes hydrogen bonding. In certain instances, the solvate will be capable of isolation, for example, when one or more solvent molecules are incorporated in the crystal lattice of the crystalline solid. The solvate may comprise either a stoichiometric or nonstoichiometric amount of the solvent molecules. The term "stereoisomer" refers to isomers of identical constitution that differ in the arrangement of their atoms in space. Enantiomers and diastereomers are examples of stereoisomers.

The term "enantiomer" refers to one of a pair of molecular species that are mirror images of each other and are not superimposable. The term "diastereomer" refers to stereoisomers that are not mirror images.

The term "racemate" or "racemic mixture" refers to a composition composed of equimolar quantities of two enantiomeric species, wherein the composition is devoid of optical activity.

The symbols "R" and "S" represent the configuration of substituents around a chiral carbon atom(s). The isomeric descriptors "R" and "S" are used as described herein for indicating atom configuration(s) relative to a core molecule and are intended to be used as defined in the literature (IUP AC Recommendations 1996, Pure and Applied Chemistry, 68:2193-2222 (1996)).

The term "tautomer" refers to each of two or more isomers of a compound that exist together in equilibrium and are readily interchanged by migration of an atom or group within the molecule. The term “halogen” or “halogen atoms” or “halo” as used herein includes fluorine, chlorine, bromine, and iodine atom.

The term “5-membered heterocyclyl” refers to a mono satured or unsatured group containing one or more heteroatoms selected from N and O.

The term "(C _x -C _y ) alkyl" wherein x and y are integers, refers to a straight or branched chain alkyl group having from x to y carbon atoms. Thus, when x is 1 and y is 6, for example, the term includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl and n-hexyl.

The term "(C _x -C _y )alkylene" wherein x and y are integers, refers to a C _x -C _y alkyl radical having in total two unsatisfied valencies, such as a divalent methylene radical.

The expressions “(C _x -C _y ) haloalkyl” wherein x and y are integers, refer to the above defined “C _x -C _y alkyl” groups wherein one or more hydrogen atoms are replaced by one or more halogen atoms, which can be the same or different.

Examples of said “(C _x -C _y ) haloalkyl” groups may thus include halogenated, poly-halogenated and fully halogenated alkyl groups wherein all hydrogen atoms are replaced by halogen atoms, e.g. trifluorom ethyl.

The term “(C _x -C _y ) cycloalkyl” wherein x and y are integers, refers to saturated cyclic hydrocarbon groups containing the indicated number of ring carbon atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl.

The term “aryl” refers to mono cyclic carbon ring systems which have 6 ring atoms wherein the ring is aromatic. Examples of suitable aryl monocyclic ring systems include, for instance, phenyl.

The term "heteroaryl" refers to a mono- or bi-cyclic aromatic group containing one or more heteroatoms selected from S, N and O, and includes groups having two such monocyclic rings, or one such monocyclic ring and one monocyclic aryl ring, which are fused through a common bond. The term “(C _x -C _y ) heterocycloalkyl” wherein x and y are integers, refers to saturated or partially unsaturated monocyclic (C _x -C _y ) cycloalkyl groups in which at least one ring carbon atom is replaced by at least one heteroatom (e.g. N, S or O) or may bear an -oxo (=0) substituent group. Said heterocycloalkyl may be further optionally substituted on the available positions in the ring, namely on a carbon atom, or on an heteroatom available for substitution. Substitution on a carbon atom includes spiro di substitution as well as substitution on two adjacent carbon atoms, in both cases thus form additional condensed 5 to 6 membered heterocyclic ring.

The term “(C _x -C _y ) aminoalkyl” wherein x and y are integers, refers to the above defined “(Ci-Ce) alkyl” groups wherein one or more hydrogen atoms are replaced by one or more amino group.

The term “(C _x -C _y ) hydroxyalkyl” wherein x and y are integers, refers to the above defined “(Ci-Ce) alkyl” groups wherein one or more hydrogen atoms are replaced by one or more hydroxy (OH) group.

The term “(C _x -C _y ) alkoxy” or “(C _x -C _y ) alkoxyl” wherein x and y are integers, refer to a straight or branched hydrocarbon of the indicated number of carbons, attached to the rest of the molecule through an oxygen bridge.

A dash that is not between two letters or symbols is meant to represent the point of attachment for a substituent.

The carbonyl group is herein preferably represented as -C(O)- as an alternative to the other common representations such as -CO-, -(CO)- or -C(=0)-.

In general, the bracketed group is a lateral group, not included into the chain, and brackets are used, when deemed useful, to help disambiguating linear chemical formulas; e.g. the sulfonyl group -SO2- might be also represented as -S(0) _2- to disambiguate e.g. with respect to the sulfmic group -S(0)0-

Whenever basic amino or quaternary ammonium groups are present in the compounds of formula I, physiologically acceptable anions may be present, selected among chloride, bromide, iodide, trifluoroacetate, formate, sulfate, phosphate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, tartrate, oxalate, succinate, benzoate, p-toluenesulfonate, pamoate and naphthalene disulfonate. Likewise, in the presence of acidic groups such as COOH groups, corresponding physiological cation salts may be present as well, for instance including alkaline or alkaline earth metal ions.

As above indicated, the present invention refers to a series of compounds represented by the general formula (I) as herein below described in details, which are endowed with a dual selectivity towards receptors LPA1 and LPA2.

Advantageously, the duality of action can be effective in the treatment of those diseases where the LPA receptors play a relevant role in the pathogenesis such as fibrosis and disease, disorder and condition from fibrosis.

Differently from similar compounds of the prior art, the compounds of formula (I) of the present invention are able to act as antagonist of both LPA1 and LPA2 receptors in a substantive and effective way, particularly appreciated by the skilled person when looking at a suitable and efficacious compounds useful for the treatment of fibrosis, in particular idiopatic pulmonary fibrosis.

As indicated in the experimental part, in fact, the counpounds of formual (I) of the invention have a dual activity as shown in Table 2, wherein it is reported the potency expressed as half maximal inhibitory concentration (IC50) on both receptors.

As it can be appreciated, the compounds of the present invention according to Table 2, show a notable potency with respect to their inhibitory activity on both receptors LPA1 and LPA2 even below about 200 nM on both LPA1 and LPA2, confirming that they are able to antagonize the two isoforms of LPA receptor mainly involved in fibrosis and diseases that result from fibrosis. It has to be reminded that the selectivity towards LPA2 is a distinct activity than the selectivity on the LPA1 receptor. In other words, the dual activity on LPA1 and LPA2 is a peculiar feature of the present compounds, not suggested or derivable from the teaching of the prior art. Even more advantageously, the compounds of formula (I) of the present invention, not only are able to act as dual antagonist for the LPA1 and LPA2 receptors, but they can be easily prepared by a convenient and synthetic process as detailed herein below.

Thus, in one aspect the present invention relates to a compound of general formula (I) as dual LPA1 and LPA2 antagonist wherein Xi, X ₂ and X ₃ are independently selected from S, N, C, CH, S(0) ₂ , S(O) provided that at least one is S, S(O) or S(0) ₂ and R ₇ is present only when X ₂ and/or X ₃ are C or N; X _4, X ₅ , Xr _> and X ₇ are independently N, C or CH wherein no more than two are N;

Ri is absent when X4 is N, or is H or selected from the group consisting of (Ci-C4)alkyl, halo, CN, and -OR2;

Li is a bond or an (Ci-C4)alkylene group;

L2 1S selected from the group consisting of wherein Li and L3 indicate the attachment of L ₂ to these groups;

L ₃ is a bond or an (Cl-C6)alkylene group wherein said alkylene may be optionally substituted by one or more group selected from (Ci-C4)alkyl, -OR3, -NR4R5, -(Ci-C4)alkylene-OR ₃ , -(Ci-C4)alkylene-NR4R5, halo and oxo;

Q is H or selected from the group consisting of (C3-C8)cycloalkyl, (C3- C ₈ )heterocycloalkyl, aryl and 5-6 membered heteroaryl, wherein each of said cycloalkyl, heterocycloalkyl, aryl and heteroaryl may be optionally substituted by one or more group selected from (Ci-C4)alkyl, -NR4R5, -(Ci-C4)alkylene-OR ₃ , -(Ci-C4)alkylene-NR4R5, halo, -NR ₂ C(0)R ₆ , -NR ₂ C(0)0R ₆ , -(Ci-C ₄ )alkylene-NR ₂ C(0)R ₆ and

-(Ci-C ₄ )alkylene-NR ₂ C(0)0R ₆ ;

R2 is H or an (Ci-C ₄ )alkyl group;

Rs is H or selected from the group consisting of (Ci-C4)alkyl, (Ci-C4)haloalkyl, (C3-C8)cycloalkyl and (C3-C8)heterocycloalkyl; R4 and Rs are at each occurrence independently H or selected from the group consisting of (Ci-C4)alkyl, (C3-C8)cycloalkyl, (Ci-Ce) haloalkyl, (Ci-Ce) hydroxyalkyl, (Ci-Ce) aminoalkyl, (Ci-Ce) alkoxyl, (Ci-Ce) alkoxy-(Ci-C ₆ ) alkyl, (C3-C6) heterocycloalkyl- (C1-C6) alkyl, aryl, heteroaryl and (C3-C6) heterocycloalkyl; or

R4 and R5 may form together with the nitrogen atom to which they are attached a 5 or 6 membered saturated heterocyclic ring system optionally containing a further heteroatom selected from N, S or O, said heterocyclic ring system may be optionally substituted by one or more groups selected from (Ci-C4)alkyl, -CN, (Ci-Ce) haloalkyl, (Ci-Ce) hydroxy alkyl,

(C ₁ -C ₆ ) aminoalkyl, halogen and oxo;

Re is H or selected from the group consisting of (Ci-C ₄ )alkyl, (Ci-Ce) haloalkyl, (Ci-Ce) hydroxyalkyl, (Ci-Ce) aminoalkyl and (Ci-Ce) alkoxyl, R7 is H or selected from(Ci-C ₄ )alkyl and halo.

The invention further concerns the corresponding deuterated derivatives of compounds of formula (I).

It has to be intended for the present invention that when Li is a bond, the group L ₂ is directly linked to the 5-membered heterocycle; when L ₃ is a bond, L ₂ is directly linked to Q; when both Li and L ₃ are bond, L ₂ is directly linked to 5-membered heterocycle and to Q.

It has also to be intended for the present invention that when Ri is absent, X ₄ is N or CH.

In one preferred embodiment, the invention refers to compound of formula (I) wherein Xi is S and X ₂ and X ₃ are C;

X _4, X ₅ , Xr _> and X ₇ are independently N, C or CH wherein no more than two are N; Ri is absent when X ₄ is N or is H or selected from the group consisting of (Ci- C ₄ )alkyl, halo, CN, and -OR ₂ ;

Li is a bond or an (Ci-C ₄ )alkylene group; L2 is selected from the group consisting of wherein Li and L ₃ indicate the attachment of L ₂ to these groups;

L ₃ is a bond or an (Ci-C6)alkylene group wherein said alkylene may be optionally substituted by one or more group selected from (Ci-C ₄ )alkyl, -OR ₃ , -NR ₄ R ₅ , -(Ci-C4)alkylene-OR3, -(Ci-C4)alkylene-NR4R5, halo and oxo;

Q is H or selected from the group consisting of (C ₃ -C ₈ )cycloalkyl, (C ₃ -C ₈ )heterocycloalkyl, aryl and 5-6 membered heteroaryl, wherein each of said cycloalkyl, heterocycloalkyl, aryl and heteroaryl may be optionally substituted by one or more group selected from (Ci-C ₄ )alkyl, -NR ₄ R ₅ , -(Ci-C ₄ )alkylene-OR ₃ , -(Ci-C ₄ )alkylene- NR4R5, halo, -NR ₂ C(0)R ₆ , -NR ₂ C(0)0R ₆ , -(Ci-C ₄ )alkylene-NR ₂ C(0)R ₆ and

-(Ci-C ₄ )alkylene-NR ₂ C(0)0R ₆ ;

R2 is H or an (Ci-C ₄ )alkyl group;

Rs is H or selected from the group consisting of (Ci-C ₄ )alkyl, (Ci-C ₄ )haloalkyl, (C ₃ -C ₈ )cycloalkyl and (C ₃ -C ₈ )heterocycloalkyl;

R4 and Rs are at each occurrence independently H or selected from the group consisting of (Ci-C ₄ )alkyl, (C ₃ -C ₈ )cycloalkyl, (Ci-Ce) haloalkyl, (Ci-Ce) hydroxyalkyl, (C1-C ₆ ) aminoalkyl, (Ci-Ce) alkoxyl, (Ci-Ce) alkoxy-(Ci-C ₆ ) alkyl, (C ₃ -C ₆ ) heterocycloalkyl-(Ci-C ₆ ) alkyl, aryl, heteroaryl and (C ₃ -C ₆ ) heterocycloalkyl; or R ₄ and R ₅ may form together with the nitrogen atom to which they are attached a 5 or 6 membered saturated heterocyclic ring system optionally containing a further heteroatom selected from N, S or O, said heterocyclic ring system may be optionally substituted by one or more groups selected from (Ci-C ₄ )alkyl, -CN, (Ci-Ce) haloalkyl, (Ci-Ce) hydroxyalkyl,

(Ci-Ce) aminoalkyl, halogen and oxo;

Re is H or selected from the group consisting of (Ci-C ₄ )alkyl, (Ci-Ce) haloalkyl, (Ci-Ce) hydroxyalkyl, (Ci-Ce) aminoalkyl and (Ci-Ce) alkoxyl,

R7 is H or selected from (Ci-C ₄ )alkyl and halo. In a further preferred embodiment, the invention refers to compound of formula (I) wherein Xi, X ₂ and X ₃ are independently selected from S, N, C, CH, S(0) ₂ , S(O) provided that at least one is S, S(O) or S(0) ₂ and R ₇ is present only when X ₂ and/or X ₃ are C or N; X4 is C and X5, Xe and X7 are CH; Ri is H or selected from the group consisting of (Ci-C4)alkyl, halo, CN, and -OR2;

Li is a bond or an (Ci-C ₄ )alkylene group; L21S selected from the group consisting of wherein Li and L3 indicate the attachment of L2 to these groups; L ₃ is a bond or an (Ci-C ₆ )alkylene group wherein said alkylene may be optionally substituted by one or more group selected from (Ci-C4)alkyl, -OR3, -NR4R5, -(Ci-C4)alkylene-OR ₃ , -(Ci-C4)alkylene-NR4R5, halo and oxo;

Q is H or selected from the group consisting of (C3-C8)cycloalkyl, (C3- C ₈ )heterocycloalkyl, aryl and 5-6 membered heteroaryl, wherein each of said cycloalkyl, heterocycloalkyl, aryl and heteroaryl may be optionally substituted by one or more group selected from (Ci-C4)alkyl, -NR4R5, -(Ci-C4)alkylene-OR ₃ , -(Ci-C4)alkylene-NR4R5, halo, -NR ₂ C(0)R ₆ , -NR ₂ C(0)0R ₆ , -(Ci-C4)alkylene-NR ₂ C(0)R ₆ and

-(Ci-C4)alkylene-NR ₂ C(0)0R ₆ ;

R2 is H or an (Ci-C ₄ )alkyl group; R ₃ is H or selected from the group consisting of (Ci-C ₄ )alkyl, (Ci-C ₄ )haloalkyl, (C ₃ -C ₈ )cycloalkyl and (C ₃ -C ₈ )heterocycloalkyl; R4 and R5 are at each occurrence independently H or selected from the group consisting of (Ci-C ₄ )alkyl, (C ₃ -C ₈ )cycloalkyl, (Ci-Ce) haloalkyl, (Ci-Ce) hydroxyalkyl, (Ci-Ce) aminoalkyl, (Ci-Ce) alkoxyl, (Ci-Ce) alkoxy-(Ci-C6) alkyl, (C3-C6) heterocycloalkyl-(Ci-C6) alkyl, aryl, heteroaryl and (C3-C6) heterocycloalkyl; or R ₄ and R ₅ may form together with the nitrogen atom to which they are attached a 5 or 6 membered saturated heterocyclic ring system optionally containing a further heteroatom selected from N, S or O, said heterocyclic ring system may be optionally substituted by one or more groups selected from (Ci-C ₄ )alkyl, -CN, (Ci-Ce) haloalkyl, (Ci-Ce) hydroxyalkyl,

(Ci-Ce) aminoalkyl, halogen and oxo;

Re is H or selected from the group consisting of (Ci-C ₄ )alkyl, (Ci-Ce) haloalkyl, (Ci-Ce) hydroxyalkyl, (Ci-Ce) aminoalkyl and (Ci-Ce) alkoxyl;

R7 is H or selected from (Ci-C ₄ )alkyl and halo.

In a further preferred embodiment, the invention refers to compound of formula (I) wherein Xi is S, X ₅ , Xr, and X ₇ are CH, X ₄ , X ₂ and X ₃ are C;

Ri is H;

Li is a bond;

L21S wherein Li and L ₃ indicate the attachment of L ₂ to these groups;

R2 is H;

L3 is (Ci-C ₄ )alkylene wherein said alkylene is optionally substituted by (Ci-C ₄ )alkyl, preferably methyl,

Q is aryl, wherein said aryl may be optionally substituted by halo, preferably chlorine, R is H or selected from (Ci-C4)alkyl and halo.

According to the above preferred embodiment, the invention refers to the compound in the Table 1 below and pharmaceutical acceptable salts thereof.

Table 1 Preferred ccompounds of Formula (I) The compounds of formula (I) including all the compounds here above listed can be generally prepared according to the procedure outlined in Schemes shown below using generally known methods. Scheme 1

Scheme 2

Scheme 3

Scheme 4

(I) Example 4 In a further aspect, the invention refers to a compound of formula V In a further aspect, the invention refers to the use of the compound V as intermediate for the preparation of Example 1.

In a further aspect, the invention refers to a compound of formula XI In a further aspect, the invention refers to the use of the compound XI as intermediate for the preparation of Example 2.

In a further aspect, the invention refers to a compound of formula XVI In a further aspect, the invention refers to the use of the compound XVI as intermediate for the preparation of Example 3.

In a further aspect, the invention refers to a compound of formula XIX

In a further aspect, the invention refers to the use of the compound XIX as intermediate for the preparation of Example 4.

The compounds of formula (I) of the present invention have surprisingly been found to effectively inhibit both receptors LPA1 and LPA2. Advantageously, the inhibition of both receptors LPA1 and LPA2 may result in a more efficacious treatment of the diseases or condition wherein the LPA receptors are involved.

In particular in this respect, it has now been found that the compounds of formula (I) of the present invention have an antagonist drug potency expressed as half maximal inhibitory concentration (IC ₅₀ ) on LPA1 and LPA2 lesser or equal than 200 nM as shown in the present experimental part.

Preferably, the compounds of the present invention have an IC50 on LPA1 and LPA2 lesser or equal than 100 nM. More preferably, the compounds of the present invention have an IC50 on LPA1 and LPA2 lesser or equal than 45 nM.

In one aspect, the present invention refers to a compound of formula (I) or a stereoisomer, tautomer or pharmaceutically acceptable salt or solvate thereof for use as a medicament. In a preferred embodiment, the invention refers to a compound of formula (I) or a stereoisomer, tautomer or pharmaceutically acceptable salt or solvate thereof, for use in the treatment of disorders associated with LPA receptors mechanism.

In a further embodiment, the present invention refers to a compound of formula (I) for use in the treatment of a disease, disorder or condition associated with dysregulation of lysophosphatidic acid receptor 1 (LPA1) and receptor 2 (LPA2).

In one embodiment, the present invention refers to a compound of formula (I) useful for the prevention and/or treatment of fibrosis and/or diseases, disorders, or conditions that involve fibrosis.

The terms "fibrosis" or "fibrosing disorder," as used herein, refers to conditions that are associated with the abnormal accumulation of cells and/or fibronectin and/or collagen and/or increased fibroblast recruitment and include but are not limited to fibrosis of individual organs or tissues such as the heart, kidney, liver, joints, lung, pleural tissue, peritoneal tissue, skin, cornea, retina, musculoskeletal and digestive tract.

Preferably, the compounds of formula (I) of the present invention are useful for the treatment and/or prevention of fibrosis such as pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), hepatic fibrosis, renal fibrosis, ocular fibrosis, cardiac fibrosis, arterial fibrosis and systemic sclerosis.

More preferably, the compounds of formula (I) of the present invention are useful for the treatment of idiopathic pulmonary fibrosis (IPF).

Advantageously, the duality of action can be particularly efficacious in the treatment of those diseases where the LPA1 and LPA2 receptors play a relevant role in the pathogenesis such as fibrosis and disease, disorder and condition from fibrosis, and more in particular for the treatment of IPF.

In one aspect, the invention also refers to a method for the prevention and/or treatment of disorders associated with LPA receptors mechanisms, said method comprises administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula (I).

In one aspect, the invention refers to the use of a compound of formula (I) in the preparation of a medicament for the treatment of disorders associated with LPA receptors mechanism.

In a further aspect, the invention refers to a method for the prevention and/or treatment of disorder or condition associated with dysregulation of lysophosphatidic acid receptor 1 (LPA1) and receptor 2 (LPA2) administering a patient in need of such treatment a therapeutically effective amount of a compound of formula (I). In a further aspect, the invention refers to the use of a compound of formula (I) according to the invention, or a pharmaceutically acceptable salt thereof, for the treatment of disorders associated with LPA receptors mechanism.

In a further aspect, the present invention refers to the use of a compound of formula (I) for the treatment of a disease, disorder or condition associated with dysregulation of lysophosphatidic acid receptor 1 (LPA1) and receptor 2 (LPA2).

As used herein, "safe and effective amount" in reference to a compound of formula (I) or a pharmaceutically acceptable salt thereof or other pharmaceutically-active agent means an amount of the compound sufficient to treat the patient's condition but low enough to avoid serious side effects and it can nevertheless be routinely determined by the skilled artisan.

The compounds of formula (I) or pharmaceutically acceptable salts thereof may be administered once or according to a dosing regimen wherein a number of doses are administered at varying intervals of time for a given period of time. Typical daily dosages may vary depending upon the route of administration chosen. The present invention also refers to a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof in admixture with at least one or more pharmaceutically acceptable carrier or excipient.

In one embodiment, the invention refers to a pharmaceutical composition of compounds of formula (I) in admixture with one or more pharmaceutically acceptable carrier or excipient, for example those described in Remington’s Pharmaceutical Sciences Handbook, XVII Ed., Mack Pub., N.Y., U.S.A.

Administration of the compounds of the invention and their pharmaceutical compositions may be accomplished according to patient needs, for example, orally, nasally, parenterally (subcutaneously, intravenously, intramuscularly, intrastemally and by infusion) and by inhalation.

Preferably, the compounds of the present invention are administered orally or by inhalation.

More preferably, the compounds of the present invention are administered orally.

In one preferred embodiment, the pharmaceutical composition comprising the compound of formula (I) is a solid oral dosage form such as tablets, gelcaps, capsules, caplets, granules, lozenges and bulk powders.

In one embodiment, the pharmaceutical composition comprising the compound of formula (I) is a tablet.

The compounds of the invention can be administered alone or combined with various pharmaceutically acceptable carriers, diluents (such as sucrose, mannitol, lactose, starches) and known excipients, including suspending agents, solubilizers, buffering agents, binders, disintegrants, preservatives, colorants, flavorants, lubricants and the like.

In a further embodiment, the pharmaceutical composition comprising a compound of formula (I) is a liquid oral dosage forms such as aqueous and non-aqueous solutions, emulsions, suspensions, syrups, and elixirs. Such liquid dosage forms can also contain suitable known inert diluents such as water and suitable known excipients such as preservatives, wetting agents, sweeteners, flavorants, as well as agents for emulsifying and/or suspending the compounds of the invention.

In a further embodiment, the pharmaceutical composition comprising the compound of formula (I) is an inhalable preparation such as inhalable powders, propellant-containing metering aerosols or propellant-free inhalable formulations.

For administration as a dry powder, single- or multi-dose inhalers known from the prior art may be utilized. In that case the powder may be filled in gelatine, plastic or other capsules, cartridges or blister packs or in a reservoir. A diluent or carrier chemically inert to the compounds of the invention, e.g. lactose or any other additive suitable for improving the respirable fraction may be added to the powdered compounds of the invention.

Inhalation aerosols containing propellant gas such as hydrofluoroalkanes may contain the compounds of the invention either in solution or in dispersed form. The propellant-driven formulations may also contain other ingredients such as co-solvents, stabilizers and optionally other excipients.

The propellant-free inhalable formulations comprising the compounds of the invention may be in form of solutions or suspensions in an aqueous, alcoholic or hydroalcoholic medium and they may be delivered by jet or ultrasonic nebulizers known from the prior art or by soft-mist nebulizers.

The compounds of the invention can be administered as the sole active agent or in combination with other pharmaceutical active ingredients.

The dosages of the compounds of the invention depend upon a variety of factors including among others the particular disease to be treated, the severity of the symptoms, the route of administration and the like. The invention is also directed to a device comprising a pharmaceutical composition comprising a compound of Formula (I) according to the invention, in form of a single- or multi-dose dry powder inhaler or a metered dose inhaler.

All preferred groups or embodiments described above for compounds of formula I may be combined among each other and apply as well mutatis mutandis.

The various aspects of the invention described in this application are illustrated by the following examples which are not meant to limit the invention in any way.

PREPARATIONS OF INTERMEDIATES AND EXAMPLES

Chemical names were generated with Chem Draw version 18.2 using the embedded Structure to Name tool, Copyright of Perkin Elmer Informatics, Inc.

In some cases, generally accepted names of commercially available reagents were used in place of Dotmatics software generated names.

All reagents, for which the synthesis is not described in the experimental part, are either commercially available, or are known compounds or may be formed from known compounds by known methods by a person skilled in the art.

ABBREVIATION - MEANING AcOK: potassium acetate ADDP: 1 T-(azodicarbonyl)dipiperidine BOC: tert butoxy carbonyl DBAD: di-tert-butyl azodicarboxylate DCM: dichloromethane DIPEA: diisopropyl ethyl amine DMAP: dimethylaminopyridine DMF: dimethylformamide Et20: diethyl ether EtOAc: ethyl acetate H2O2: hydrogen peroxide HC1: hydrochloric acid HCOOH: formic acid K2CO3: potassium carbonate KHSO4: potassium hydrogen sulfate KOH: potassium hydroxide

LC/MS: Liquid chromatography/mass spectrometry

LiOH: lithium hydroxide

Mel: iodomethane

MeOH: methanol

MgSCL: magnesium sulfate

Na ₂ CC> ₃ : sodium carbonate

Na2S2C> ₃ : Sodium thiosulfate

Na ₂ SC> ₄ : sodium sulfate

NaCl: sodium chloride

NH ₄ CI: ammonium chloride

Pd(dppf)Ch: 1 l'-bis(diphenylphosphino)ferrocene dichloropalladium (II)

PPh3 : triphenylphosphine pTLC: preparative thin layer chromatography

PTSA: p-toluenesulfonic acid

TEA: triethylamine

THF : tetrahydrofuran

General Experimental Details and methods

Analytical method

NMR characterization 1H-NMR spectra were performed on a Varian MR-400 spectrometer operating at 400 MHZ (proton frequency), equipped with: a self-shielded Z-gradient coil 5 mm lH/nX broadband probe head for reverse detection, deuterium digital lock channel unit, quadrature digital detection unit with trans mitter offset frequency shift, or on AgilentVNMRS-500 or on a Bruker Avance 400 spectrometers. Chemical shift are reported as 6 values in ppm relative to trimethylsilane (TMS) as an internal standard. Coupling constants (J values) are given in hertz (Hz) and multiplicities are reported using the following abbreviation (s=singlet, d=doublet, t=triplet, q=quartet, m=multiplet, br. s=broad, nd=not determined). LC/UV/MS Analytical Method

UPLC/MS (ES+/ES-) was performed on AcquityTM system coupled a Waters SQD mass spectrometer using a column Acquity UPLC CSH Cl 8 column (50mm x 2.1mm i.d. 1.7 pm particle size) with a column Temperature (°C) 40.0 and a mobile phases: 0.1% v/v solution of HCOOH in water (A); 0.1% v/v solution of HCOOH in Acetonitrile (B). Flow (ml/min) 1

Stop Time (mins) 2.0 Gradient:

UV Conditions:

UV detection range: 210 nm to 350 nm Acquisition rate: 40 Hz

DAD - MS Rt offset: 0.01 min MS Conditions:

Ionization mode: alternate Positive/Negative Electrospray (ES+/ES-)

Scan Range: 100 to 1000 AMU

Example 1

Synthesis of (IS, 3S)-3-[4-[3-[[(lR)-l-(2- chlorophenyl)ethoxy]carbonylamino]thiophen-2-yl]phenoxy]cycl ohexane-l- carboxylic acid

Step 1: Synthesis of ethyl 2-[4-(methoxymethoxy)phenyl]thiophene-3- carboxylate (intermediate 1) intermediate 1

A mixture of ethyl 2-bromothiophene-3-carboxylate (542.51 mg, 2.31 mmol), [4- (methoxymethoxy)phenyl]boronic acid (350 mg, 1.92 mmol), sodium carbonate (681 mg, 5.76 mmol) in 1,2-dimethoxy ethane (12 mL) and Water (4 mL) was degassed by applying alternatively vacuum and nitrogen. [1,1'- Bis(diphenylphosphino)ferrocene]dichloropalladium(II) (0.07 g, 0.095 mmol) was then added and the mixture was heated at 80 °C. After lh an UPLC/MS check showed complete conversion to the desired compound. The mixture was then diluted with EtOAc, washed with water and brine, dried over Na2S04, filtered and concentrated in vacuo to give a crude which was purified by FC (Si SNAP 50, CyHex/AcOEt from 10/0 to 9/1) to afford ethyl 2-[4-(methoxymethoxy)phenyl]thiophene-3-carboxylate (489 mg, 1.673 mmol, 87.97% yield) as colorless oil. LC-MS (ESI): m/z (M+l): 293.18.

¾NMR (400 MHz, DMSO-d6) ppm: 1.15 (t, J=7.15 Hz, 3 H) 3.35 - 3.46 (m, 3 H) 4.14 (q, J=7.04 Hz, 2 H) 5.18 - 5.31 (m, 2 H) 7.07 (d, J=8.80 Hz, 2 H) 7.38 - 7.45 (m, 3 H) 7.58 (d, J=5.50 Hz, 1 H).

Step 2: Synthesis of 2-[4-(methoxymethoxy)phenyl]thiophene-3-carboxylic acid (intermediate 2) intermediate 2 Ethyl 2-[4-(methoxymethoxy)phenyl]thiophene-3-carboxylate (489.0 mg, 1.44 mmol) was dissolved in THF (3 mL), Water (3 mL) and Methanol (3 mL). lithium hydroxide (68.9 mg, 2.88 mmol) was then added and the mixture was stirred at rt overnight. The solvents were removed under vacuum, then HC1 1 M was added to slightly acidic pH and the mixture was extracted with ethyl acetate (3x). The mixed organic phases were washed with brine, dried over Na2S04, filtered and concentrated in vacuo to give 2-[4-(methoxymethoxy)phenyl]thiophene-3-carboxylic acid (400.7 mg, 1.516 mmol, quantitative yield) as white solid, which was used in the next step without further purification.

LC-MS (ESI): m/z (M+l): 265.07. ¾ NMR (400 MHz, DMSO-d6) ppm: 3.34 - 3.45 (m, 3 H) 5.24 (s, 2 H) 7.06 (d,

J=8.80 Hz, 2 H) 7.36 - 7.46 (m, 3 H) 7.54 (d, J=5.50 Hz, 1 H) 12.35 - 12.82 (m, 1 H). Step 3: Synthesis of [(lR)-l-(2-chlorophenyl)ethyl] N-[2-[4-

(methoxymethoxy)phenyl]thiophen-3-yl]carbamate (intermediate 3) intermediate 3

A solution of (R)-l-(2-chlorophenyl)ethanol (0.24 mL, 1.82 mmol) in dry Toluene (15 mL) was added to 2-[4-(methoxymethoxy)phenyl]thiophene-3-carboxylic acid (400.7 mg, 1.52 mmol) Triethylamine (0.42 mL, 3.03 mmol) and [azido(phenoxy)phosphoryl]oxybenzene (0.49 mL, 2.27 mmol) were added and the mixture was heated at 125 °C. An UPLC/MS check after 2 h showed complete conversion. The mixture was then diluted with ethyl acetate and washed with a saturated aqueous NaHC03 solution (2x) and brine. The organic phase was dried over Na2S04, filtered and concentrated in vacuo to give a crude material which was purified by flash chromatography (Biotage KP-Sil lOOg SNAP cartridge, gradient cyclohexane/ethyl acetate from 10/0 to 8/2) to give [(lR)-l-(2-chlorophenyl)ethyl] N-[2-[4- (methoxymethoxy)phenyl]thiophen-3-yl]carbamate (546.2 mg, 1.307 mmol, 86.21% yield) as yellow oil.

LC-MS (ESI): m/z (M+l): 418.27.

¾ NMR (400 MHz, DMSO-d6) ppm: 1.25 - 1.38 (m, 1 H) 1.31 (d, J=6.38 Hz, 2 H) 1.41 - 1.55 (m, 2 H) 3.40 (s, 3 H) 4.97 - 5.10 (m, 1 H) 5.23 (s, 2 H) 5.30 - 5.41 (m, 1 H) 5.91 - 6.04 (m, 1 H) 7.07 (d, J=8.80 Hz, 3 H) 7.17 - 7.67 (m, 12 H) 7.22 - 7.62 (m, 11 H) 9.09 - 9.28 (m, 1 H).

Step 4: Synthesis of [(lR)-l-(2-chlorophenyl)ethyl] N-[2-(4- hydroxyphenyl)thiophen-3-yl]carbamate (intermediate 4)

[(lR)-l-(2-chlorophenyl)ethyl] N-[2-[4-(methoxymethoxy)phenyl]thiophen-3- yljcarbamate (546.2 mg, 1.31 mmol) was dissolved in THF (5.462 mL). A solution of Hydrogen chloride (10.05 mL, 6.53 mmol) (2% w/w, about 0.65M) in isopropyl alcohol was then added and the mixture was stirred at room temperature for 48 h. UPLC/MS check showed the presence of the presumed desired compound together with some residual starting material. The mixture was then neutralized with a 2N aqueous NaOH solution and concentrated in vacuo. Water was added and the mixture was extracted with ethyl acetate (3x). The mixed organic phases were dried over Na2S04, filtered and concentrated in vacuo to give a crude which was purified by FC (Si SNAP 25, CyHex/AcOEt from 10/ to 1/1) to afford [(lR)-l-(2-chlorophenyl)ethyl] N-[2-(4- hydroxyphenyl)thiophen-3-yl]carbamate (250 mg, 0.669 mmol, 51.17% yield) as white solid.

LC-MS (ESI): m/z (M+l): 374.16.

¾ NMR (400 MHz, DMSO-d6) ppm: 1.22 - 1.62 (m, 3 H) 5.84 - 6.06 (m, 1 H) 6.80 (d, J=8.80 Hz, 2 H) 7.02 (d, J=5.28 Hz, 1 H) 7.22 - 7.61 (m, 7 H) 8.95 - 9.19 (m, 1

H) 9.53 - 9.87 (m, 1 H). Step 5: Synthesis of methyl (IS, 3S)-3-[4-[3-[[(lR)-l-(2- chlorophenyl)ethoxy]carbonylamino]thiophen-2-yl]phenoxy]cycl ohexane-l-carboxylate (intermediate 5) intermediate 5

Triphenylphosphine (701.59 mg, 2.67 mmol) was dissolved in dry THF (2 mL). The mixture was cooled down to 0 °C and a solution of (NE)-N-[(2-methylpropan-2- yl)oxy-oxomethyl]iminocarbamic acid tert-butyl ester (615.92 mg, 2.67 mmol) (DBAD) in dry THF (2 mL) was added dropwise. The mixture was stirred at 0 °C. After 5min the formation of an abundant white solid was observed. After 30min a solution of methyl (lS,3R)-3-hydroxycyclohexane-l-carboxylate (190.41 mg, 1.2 mmol) and [(lR)-l-(2- chlorophenyl)ethyl] N-[2-(4-hydroxyphenyl)thiophen-3-yl]carbamate (250.0 mg, 0.670 mmol) in dry THF (4 mL) was added dropwise. The mixture was then stirred at 0 °C for 30 min, then at rt 30 min and at 60 °C overnight. Water was then added and the mixture was extracted with ethyl acetate (3x). The mixed organic phases were dried over Na2S04, filtered and concentrated in vacuo to give a crude which was purified by flash chromatography (Biotage KP-Sil 25g SNAP cartridge, gradient CyHex/AcOEt from 10/0 to 1/1,) to afford methyl (lS,3S)-3-[4-[3-[[(lR)-l-(2- chlorophenyl)ethoxy]carbonylamino]thiophen-2-yl]phenoxy]cycl ohexane-l-carboxylate (44.5 mg, 0.087 mmol, 12.95% yield) as colorless oil.

LC-MS (ESI): m/z (M+l): 514.38. ¾ NMR (400 MHz, DMSO-d6) ppm: 1.51 (s, 13 H) 1.14 - 1.69 (m, 12 H) 1.73 - 1.92 (m, 3 H) 1.93 - 2.03 (m, 1 H) 2.71 - 2.82 (m, 1 H) 3.62 (s, 3 H) 4.61 - 4.78 (m, 1 H) 5.86 - 6.04 (m, 1 H) 6.90 - 7.13 (m, 3 H) 7.24 - 7.69 (m, 7 H) 9.06 - 9.24 (m, 1 H).

Step 6: Synthesis of (IS, 3S)-3-[4-[3-[[(lR)-l-(2- chlorophenyl)ethoxy]carbonylamino]thiophen-2-yl]phenoxy]cycl ohexane-l -carboxylic acid (Example 1) Example 1

To a solution of methyl (lS,3S)-3-[4-[3-[[(lR)-l-(2- chlorophenyl)ethoxy]carbonylamino]thiophen-2-yl]phenoxy]cycl ohexane-l-carboxylate (44.5 mg, 0.090 mmol) in THF (1 mL), Water (1 mL) and Methanol (1 mL), lithium hydroxide (4.15 mg, 0.170 mmol) was added and the mixture was stirred at rt 2 h . The solvents were removed under vacuum, then HC1 1 M was added to acidic pH and the mixture was extracted with ethyl acetate (3x). The mixed organic phases were washed with brine, dried over Na2S04, filtered and concentrated in vacuo to give a crude which was purified by FC (Si SNAP lOg, DCM/MeOH from 10/0 to 95/5) to afford a first crude which was further purified by FC reverse phase (Si C18 12 g, water with 0.1% HCOOH/MeCN from 10/0 to 30/60) to obtain (lS,3S)-3-[4-[3-[[(lR)-l-(2- chlorophenyl)ethoxy]carbonylamino]thiophen-2-yl]phenoxy]cycl ohexane-l -carboxylic acid (20 mg, 0.040 mmol, 46.2% yield) as white solid.

LC-MS (ESI): m/z (M+l): 500.35. ¾NMR (400 MHz, DMSO-d6) ppm: 1.28 - 1.97 (m, 11 H), 2.55 - 2.65 (m, 1 H), 4.67 (br s, 1 H), 5.83 - 6.02 (m, 1 H), 6.84 - 7.10 (m, 3 H), 7.15 - 7.81 (m, 7 H), 8.87 - 9.35 (m, 1 H), 11.53 - 13.10 (m, 1 H).

Example 2 Synthesis of (lS,3S)-3-{4-[5-chloro-3-({[(lR)-l-(2- chlorophenyl)ethoxy]carbonyl}amino)thiophen-2-yl]phenoxy}cyc lohexane-l- carboxylic acid

Step 1: Preparation of ethyl 2-bromo-5-chlorothiophene-3-carboxylate (Intermediate 6) intermediate 6 To a solution of ethyl 2-bromothiophene-3-carboxylate (300 mg, 1.28 mmol) in DMF (8 mL), l-chloropyrrolidine-2,5-dione (511 mg, 3.8 mmol) was added and the mixture was stirred at 60 °C for 5h. NaHCCb sat. sol. and AcOEt were added and the mixture was extracted with AcOEt, collected organic phases were dried over Na ₂ S0 ₄ , filtered and evaporated. The crude was purified by FC (Si SNAP 25, CyHex/ AcOEt from 10/0 to 8/2) to afford title compound (345 mg, 1.28 mmol, quantitative yield) as a colorless oil.

LC-MS (ESI): m!z (M+l): 271 ¾ NMR (400 MHz, DMSO-r¾) d ppm 1.30 (t, 7=7.15 Hz, 3 H) 4.28 (d, 7=7.04 Hz, 2 H) 7.39 (s, 1 H)

Step 2: Preparation of ethyl 5-chloro-2-[4-(methoxymethoxy)phenyl]thiophene-3- carboxylate (Intermediate 7) intermediate 7

Title compound was prepared following the procedure used for the synthesis of Intermediate 1, starting from ethyl 2-bromo-5-chlorothiophene-3-carboxylate (Intermediate 6, 345 mg, 1.28 mmol), to afford title compound (251 mg, 0.76 mmol, 60% yield) as a colorless oil. LC-MS (ESI): mlz (M+l): 327.2

¾ NMR (400 MHz, DMSO^) d ppm 1.12 - 1.16 (m, 3 H) 3.40 (s, 3 H) 4.13 - 4.15 (m, 2 H) 5.25 (s, 2 H) 7-07 - 7.09 (m, 2 H) 7.30 - 7.52 (m, 3 H)

Step 3: Preparation of 5-chloro-2-[4-(methoxymethoxy)phenyl]thiophene-3- carboxylic acid (Intermediate 8) intermediate 8

Title compound was prepared following the procedure used for the synthesis of Intermediate 2, starting from ethyl 5-chloro-2-[4-(methoxymethoxy)phenyl]thiophene-3- carboxylate (Intermediate 7, 251 mg, 0.76 mmol), to afford title compound (214 mg, 0.72 mmol, 93% yield) as a white solid.

LC-MS (ESI): mlz (M+l): 299.1

¾ NMR (400 MHz, DMSO-r¾) d ppm 3.40 (s, 3 H) 5.25 (s, 2 H) 7.06 (d, J=8.80 Hz, 2 H) 7.38 (s, 1 H) 7.44 (d, J=8.80 Hz, 2 H) 12.84 (br. s., 1 H)

Step 4: Preparation of [(lR)-l-(2-chlorophenyl)ethyl] N-[5-chloro-2-[4- (methoxymethoxy)phenyl] thiophen-3-yl]carbamate (Intermediate 9)

Title compound was prepared following the procedure used for the synthesis of Intermediate 3, starting from 5-chloro-2-[4-(methoxymethoxy)phenyl]thiophene-3- carboxylic acid (Intermediate 8, 214 mg, 0.72 mmol), to afford title compound (329 mg, 0.72 mmol, quntitative yield) as a yellow oil.

LC-MS (ESI): mlz (M+l): 452.3

Step 5: Preparation of [(lR)-l-(2-chlorophenyl)ethyl] N-[5-chloro-2-(4- hydroxyphenyl)thiophen-3-yl]carbamate (Intermediate 10) Title compound was prepared following the procedure used for the synthesis of Intermediate 4, starting from [(lR)-l-(2-chlorophenyl)ethyl] N-[5-chloro-2-[4- (methoxymethoxy)phenyl] thiophen-3-yl]carbamate (Intermediate 9, 329 mg, 0.72 mmol), to afford title compound (144 mg, 0.35 mmol, 49% yield) as a white solid. LC-MS (ESI): mlz (M+l): 408.1

Step 6: Preparation of methyl (lS,3S)-3-[4-[5-chloro-3-[[(lR)-l-(2- chlorophenyl)ethoxy] carbonylamino]thiophen-2-yl]phenoxy]cy clohexane- 1 - carboxylate (Intermediate 11) intermediate 11

Title compound was prepared following the procedure used for the synthesis of Intermediate 5, starting from [(lR)-l-(2-chlorophenyl)ethyl] N-[5-chloro-2-(4- hydroxyphenyl)thiophen-3-yl]carbamate (Intermediate 10, 77 mg, 0.189 mmol), to afford title compound (36 mg, 0.06 mmol, 35% yield) as a yellow solid. LC-MS (ESI): mlz (M+l): 548.4

Step 7: Preparation of (IS, 3S)-3-[4-[3-[[(lR)-l-(2- chlorophenyl)ethoxy]carbonylamino]thiophen-2-yl]phenoxy]cycl ohexane-l -carboxylic acid (Example 2) Example 2

Title compound was prepared following the procedure used for the synthesis of

Example 1, starting from [(lR)-l-(2-chlorophenyl)ethyl] N-[5-chloro-2-(4- hydroxyphenyl)thiophen-3-yl]carbamate (Intermediate 11, 36 mg, 0.06 mmol), to afford title compound (20 mg, 0.04 mmol, 66% yield) as a white solid.

LC-MS (ESI): mlz (M+l): 534.2

¾NMR (400 MHz, DMSO-i¾) d ppm 1.08 - 2.01 (m, 11 H), 2.53 - 2.61 (m, 1 H), 4.67 (br s, 1 H), 5.84 - 6.05 (m, 1 H), 7.01 (d, J= 8.8 Hz, 2 H), 7.09 (s, 1 H), 7.16 - 7.66 (m, 6 H), 9.34 (br s, 1 H), 12.30 (br s, 1 H)

Example 3

Synthesis of (lS,3S)-3-{4-[4-({[(lR)-l-(2-chlorophenyl)ethoxy]carbonyl}am ino)-3- methyl-l,2-thiazol-5-yl]phenoxy}cyclohexane-l-carboxylic acid (CHD-065350)

Step 1: Preparation of ethyl 5-bromo-3-methyl-l,2-thiazole-4-carboxylate (Intermediate 12) intermediate 12

A 5 mL microwave tube equipped with stirring bar was charged with a solution of 5-bromo-3-methyl-l,2-thiazole-4-carboxylic acid (500 mg, 2.25 mmol) in MeOH (4 mL). The tube was sealed and cooled in a cold water bath. Thionyl chloride (0.33 mL, 4.5 mmol) was added dropwise to the stirred solution. The mixture was heated at reflux for 4 h. The mixture was neutralized with solid NaHCCL, then evaporated under reduced pressure. The solid was suspended in EtOAc (10 mL) and water (10 mL) and the layers were separated. The aqueous phase was extracted with EtOAc (2x10 mL). The organic layers were collected, washed with brine (20 mL), dried over Na ₂ S0 ₄ , filtered and concentrated under reduced pressure to afford title compound (540 mg, crude) as a brown oil that was used in the following step without further purification.

LC-MS (ESI): mlz (M+l): 237.9

Step 2: Preparation of methyl 5-[4-(methoxymethoxy)phenyl]-3-methyl-l,2- thiazole-4-carboxylate (Intermediate 13) intermediate 13

Title compound was prepared following the procedure used for the synthesis of Intermediate 1, starting from methyl 5-bromo-3-methyl-l,2-thiazole-4-carboxylate (Intermediate 12, 540 mg, crude), to afford title compound (390 mg, 1.33 mmol, 59% yield over two steps) as a colorless oil.

LC-MS (ESI): mlz (M+l): 294.1

¾ NMR (400 MHz, Chloroform- ) d ppm 7.42 - 7.36 (m, 2H), 7.12 (d, J= 8.7 Hz, 2H), 5.25 (s, 2H), 3.78 (s, 3H), 3.53 (s, 3H), 2.68 (s, 3H)

Step 3: Preparation of 5-[4-(methoxymethoxy)phenyl]-3-methyl-l,2-thiazole-4- carboxylic acid (Intermediate 14) intermediate 14

Title compound was prepared following the procedure used for the synthesis of Intermediate 2, starting from methyl 5-[4-(methoxymethoxy)phenyl]-3-methyl-l,2- thiazole-4-carboxylate (Intermediate 13, 390 mg, 1.33 mmol), to afford title compound (340 mg, 1.22 mmol, 94% yield) as a white solid.

LC-MS (ESI): mlz (M+l): 280.1

¾ NMR (400 MHz, DMSO^) d ppm 7.56 - 7.34 (m, 2H), 7.18 - 7.08 (m, 2H), 5.26 (s, 2H), 3.40 (s, 3H), 2.55 (s, 3H)

Step 4: Preparation of (lR)-l-(2-chlorophenyl)ethyl N-{5-[4- (methoxymethoxy)phenyl]-3-methyl-l,2-thiazol-4-yl}carbamate (Intermediate 15) intermediate 15

Title compound was prepared following the procedure used for the synthesis of

Intermediate 3, starting from 5-[4-(methoxymethoxy)phenyl]-3-methyl-l,2-thiazole-4- carboxylic acid (Intermediate 14, 160 mg, 0.57 mmol), to afford title compound (220 mg, 0.51 mmol, 89% yield) as an amorphous off-white solid.

LC-MS (ESI): mlz (M+l): 433.1

¹ HNMR (400 MHz, Methanol-^) d ppm 7.57 (d, J= 7.6 Hz, 1H), 7.47 - 7.35 (m, 3H), 7.35 - 7.27 (m, 1H), 7.14 - 7.07 (m, 3H), 6.12 (q, J= 6.6 Hz, 1H), 5.25 (s, 2H), 3.49 (s, 3H), 2.33 (s, 3H), 1.58 (d, J= 6.6 Hz, 3H)

Step 5: Preparation of (lR)-l-(2-chlorophenyl)ethyl N-[5-(4-hydroxyphenyl)-3- methyl-l,2-thiazol-4-yl]carbamate (Intermediate 16)

Title compound was prepared following the procedure used for the synthesis of Intermediate 4, starting from (lR)-l-(2-chlorophenyl)ethyl N-{5-[4- (methoxymethoxy)phenyl]-3-methyl-l,2-thiazol-4-yl}carbamate (Intermediate 15, 220 mg, 0.51 mmol), to afford title compound (200 mg, 0.51 mmol, quantitative yield) as a white solid. LC-MS (ESI): m/z (M+l): 389.1

¾NMR (400 MHz, Methanol-^) d ppm 7.57 (d, J= 7.6 Hz, 1H), 7.44 - 7.30 (m, 5H), 6.85 (d, J= 8.3 Hz, 2H), 6.13 (d, J= 6.7 Hz, 1H), 2.31 (s, 3H), 1.58 (d, J= 6.5 Hz,

3H)

Step 6: Preparation of methyl (lS,3S)-3-{4-[4-({[(lR)-l-(2- chlorophenyl)ethoxy]carbonyl}amino)-3-methyl-l,2-thiazol-5- yljphenoxy} cyclohexane- 1-carboxylate (Intermediate 17) intermediate 17 Title compound was prepared following the procedure used for the synthesis of

Intermediate 5, starting from (lR)-l-(2-chlorophenyl)ethyl N-[5-(4-hydroxyphenyl)-3- methyl-l,2-thiazol-4-yl]carbamate (Intermediate 16, 100 mg, 0.26 mmol), to afford title compound (110 mg, 0.21 mmol, 81% yield).

LC-MS (ESI): m/z (M+l): 529.1 ¾ NMR (400 MHz, METHANOL-^) d ppm 7.58 (d, J= 7.6 Hz, 1H), 7.48 - 7.27

(m, 4H), 7.02 (d, J= 8.3 Hz, 3H), 6.13 (q, J= 6.7 Hz, 1H), 4.75 (hept, J= 5.3, 2.6 Hz, 1H), 3.70 (s, 3H), 3.37 (s, 2H), 2.91 - 2.80 (m, 1H), 2.32 (s, 3H), 2.13 - 2.05 (m, 1H), 1.99 - 1.86 (m, 2H), 1.84 - 1.62 (m, 4H), 1.58 (d, J= 6.6 Hz, 3H)

Step 7: Preparation of (lS,3S)-3-{4-[4-({[(lR)-l-(2- chlorophenyl)ethoxy]carbonyl}amino)-3-methyl-l,2-thiazol-5- yljphenoxy} cyclohexane- 1 -carboxylic acid (Example 3) (Example 3)

Title compound was prepared following the procedure used for the synthesis of Example 1, starting from methyl (lS,3S)-3-{4-[4-({[(lR)-l-(2- chlorophenyl)ethoxy]carbonyl}amino)-3-methyl-l,2-thiazol-5- yljphenoxy} cyclohexane- 1-carboxylate (Intermediate 17, 110 mg, 0.21 mmol), to afford title compound (65 mg, 0.126 mmol, 61% yield) as a white solid LC-MS (ESI): mlz (M+l): 515.1

¾ NMR (400 MHz, METHANOL-^ ) d ppm 1.17 - 1.58 (m, 3 H), 1.57 - 2.10 (m, 8 H), 2.25 - 2.40 (m, 3 H), 2.72 - 2.84 (m, 1 H), 4.68 - 4.78 (m, 1 H), 5.89 - 6.19 (m, 1

H), 6.74 - 7.65 (m, 1 H), 7.00 (br d, J=8.5 Hz, 2 H), 7.12 - 7.53 (m, 5 H)

Example 4

Synthesis of (1 S,3S)-3-(4-(4-((((R)-l-(2-chloropyridin-3-yl)ethoxy)carbonyl )amino)- 3-methylisothiazol-5-yl)phenoxy)cyclohexane-l-carboxylic acid Step 1: Preparation of methyl (lS,3S)-3-(4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)phenoxy)cyclohexane-l -carboxylate (Intermediate 18) intermediate 18 To a solution of triphenylphosphine (0.715 g, 2.73 mmol) in THF (6 mL) cooled to

3°C, di-tert-butyl azodi carboxyl ate (0.628 g, 2.73 mmol) was added portion-wise and the resulting mixture was stirred at the same temperature for 30 min. A solution of 4-(4,4,5,5- tetramethyl-l,3,2-dioxaborolan-2-yl)phenol (0.200 g, 0.909 mmol) and methyl (1S,3R)- 3-hydroxycyclohexane-l-carboxylate (0.575 g, 3.64 mmol) was added drop-wise at 3°C and the mixture was heated at 60°C for 3 h, then stirred at r.t. overnight. The solvent was removed under reduced pressure and the residue was purified by flash chromatography on 70 g silica Sepachrom Sphera cartridge (hexane : EtOAc = 95 : 5 to 85 : 15) to afford methyl (lS,3S)-3-(4-(4,4,5,5-tetramethyl-l,3,2-dioxaborolan-2- yl)phenoxy)cyclohexane-l -carboxylate (0.192 g, 0.533 mmol, 59% yield) as a colorless oil. LC-MS (ESI): mlz (M+l): 361.2.

Step 2: Preparation of (R)-l-(2-chloropyri din-3 -yl)ethyl (5-(4-iodophenyl)-3- methylisothiazol-4-yl)carbamate (Intermediate 19) intermediate 19 To a mixture of 5-iodo-3-methylisothiazole-4-carboxylic acid (0.200 g, 0.743 mmol) and (R)-l-(2-chloropyridin-3-yl)ethan-l-ol (0.141 g, 0.892 mmol) in toluene (15 mL), TEA (0.207 mL, 1.487 mmol) was added: the solution became clear. Diphenyl phosphoryl azide (0.240 mL, 1.115 mmol) was added and the reaction was heated at 120°C for 5 h. The mixture was cooled to r.t., diluted with EtOAc and washed with sat.

NaElCCh and with brine. The organic phase was dried over sodium sulfate, filtered and concentrated. The residue was triturated with EtOAc (4 mL). The solid was collected by filtration, washed with EtOAc and dried to afford (R)-l-(2-chloropyri din-3 -yl)ethyl (5- iodo-3-methylisothiazol-4-yl)carbamate (0.185 g, 0.437 mmol, 59% yield) as a beige solid.LC-MS (ESI): mlz (M+l): 424.2.

Step 3: Preparation of methyl (lS,3S)-3-(4-(4-((((R)-l-(2-chloropyridin-3- yl)ethoxy)carbonyl)amino)-3-methylisothiazol-5-yl)phenoxy)cy clohexane-l- carboxylate (Intermediate 20) intermediate 20

A mixture of (R)-l-(2-chloropyridin-3-yl)ethyl (5-iodo-3-methylisothiazol-4- yl)carbamate (0.093 g, 0.220 mmol), methyl (lS,3S)-3-(4-(4,4,5,5-tetramethyl-l,3,2- dioxaborolan-2-yl)phenoxy)cyclohexane-l-carboxylate (0.095 g, 0.264 mmol), sodium carbonate monohydrate (0.054 g, 0.440 mmol) in dioxane (7 mL) and water (2 mL) was degassed under nitrogen stream. PdCl2(dppf) (0.032 g, 0.044 mmol) was added and the mixture was heated at 100°C for 5 h. The mixture was cooled to r.t. and dioxane was evaporated under reduced pressure. The residue was taken up with EtOAc and water and the aqueous phase was saturated with NaCl. The organic phase was separated, dried over sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography on 10 g silica Biotage Sfar cartridge (DCM to DCM : acetone = 87 : 13) to afford methyl (lS,3S)-3-(4-(4-((((R)-l-(2-chloropyridin-3- yl)ethoxy)carbonyl)amino)-3-methylisothiazol-5-yl)phenoxy)cy clohexane-l- carboxylate (0.038 g, 0.072 mmol, 33% yield) which was used in following step without any additional purification. LC-MS (ESI): mlz (M+l): 530.3.

Step 4: Preparation of (lS,3S)-3-(4-(4-((((R)-l-(2-chloropyridin-3- yl)ethoxy)carbonyl)amino)-3-methylisothiazol-5-yl)phenoxy)cy clohexane-l -carboxylic acid (Example 4) (Example 4) To a solution of methyl (lS,3S)-3-(4-(4-((((R)-l-(2-chloropyridin-3- yl)ethoxy)carbonyl)amino)-3-methylisothiazol-5-yl)phenoxy)cy clohexane-l- carboxylate (0.037 g, 0.070 mmol) in THF (1 mL) cooled to 0°C, aqueous 1M LiOH (0.279 mL, 0.279 mmol) was added and the resulting mixture was left to warm to r.t. and stirred overnight. The mixture was cooled with an ice bath, acidified with 1M HC1 (pH 2) and extracted with EtOAc. The organic phase was washed with brine, dried over sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography on 4 g silica Sepachrom cartridge (DCM to DCM : MeOH = 97 : 3) to afford (lS,3S)-3-(4-(4-((((R)-l-(2-chloropyridin-3-yl)ethoxy)carbon yl)amino)-3- methylisothiazol-5-yl)phenoxy)cyclohexane-l -carboxylic acid (0.012 g, 0.023 mmol, 33% yield) as a white foam.

LC-MS (ESI): m/z (M+l): 516.2.

¾NMR (300 MHz, DMSO-d ₆ ) d ppm 12.21 (br s, 1H), 9.38 - 8.89 (m, 1H), 8.43 - 8.24 (m, 1H), 8.02 - 7.89 (m, 1H), 7.61 - 7.39 (m, 3H), 7.06 (d, J= 8.5 Hz, 2H), 5.94 - 5.72

(m, 1H), 4.86 - 4.67 (m, 1H), 2.72 - 2.59 (m, 1H), 2.36 - 2.16 (m, 3H), 2.02 - 1.13 (m, 11H).

PHARMACOLOGICAL ACTIVITY OF THE COMPOUNDS OF THE INVENTION In vitro Assays

The effectiveness of compounds of the present invention as dual LPA1 and LPA2 antagonist can be determined at the human recombinant LPA1 or LPA2 expressed in CHO cells, using a FLIPR assay in 384 well format.

CHO-hLPAl and hLPA2 cell lines are cultured in a humidified incubator at 5% C02 in DMEM/F-12 (1:1) MIXTURE with 2mM Glutamax, supplemented with 10% of

Foetal Bovine Serum, 1 mM Sodium Pyruvate, 11 mM Hepes and IX Penicillin/Streptomycin. CHO hLPAl or hLPA2 cells are seeded into black walled clear- bottom 384-well plates (#781091, Greiner Bio-One GmbH) at a density of 7,500 cells per well in 50 mΐ culture media and grown overnight in a 37°C humidified C02-incubator. Serial dilutions (1 :3 or 1:4, 11 points CRC) of compounds are performed in 100% DMSO at 200X the final concentration. The compounds are diluted 1:50 prior to the experiment with Assay Buffer (20 mM HEPES, 145 mM NaCl, 5 mM KC1, 5.5 mM glucose, 1 mM MgC12 and 2 mM CaC12, pH 7.4 containing 0.01% Pluronic F-127) to obtain a solution corresponding to 5-fold the final concentration in the assay (4X, 2% DMSO). The final concentration of DMSO in the assay will be 0.5% in each well. Medium is removed by aspiration and cells are then incubated with 30 mΐ of a loading solution containing 5 mM of the cytoplasmic Ca2+ indicator Cal-520 AM in Assay Buffer containing 2.5 mM probenecid for 30 min at 37°C incubator (cell loading). The loaded cell plates are transferred into the FLIPR instrument and calcium responses are monitored during the on-line addition protocols. For testing of compounds, after the cell loading, 10 mΐ/well of 4X antagonists’ solution was added onto the cells. After 30 min pre-incubation (at 37°C), 10 mΐ/well of 5X concentrated LPA EC80 was added and Ca2+ mobilization responses was followed during the on-line addition protocol. Intracellular peak fluorescence values subtracted by baseline fluorescence are exported and analysed to determine IC50 values, respectively. The calcium response is expressed as percentage of the maximal inhibition of the EC80 agonist response.

The raw data obtained in unstimulated controls (DMSO, no LPA) are set as “100% inhibition”, while the raw data obtained in negative controls, i.e. in the absence of compounds and stimulating with LPA EC80, are set as “0% inhibition”.

The raw data (peak height expressed as relative fluorescence units) are normalized and transformed into “percent of inhibition”. Curve fitting and pIC o (-LogICso) estimations are carried out using a four-parameter logistic model using XLfit Software.

The results for individual compounds are provided below in Table 2 wherein the compounds are classified in term of potency with respect to their inhibitory activity on LPA1 and LPA2 isoforms: As it can be appreciated, the compounds of Table 2 show a good dual activity as antagonist of both LPA1 and LPA2.

Table 2 LP A receptor 1 (LPA1)

+: LPA1 IC50 comprised between about 100 nM and 200 nM ++: LPA1 IC50 comprised between about 45 nM and 100 nm +++: LPA1 IC50 less than about 45 nM.

LPA receptor 2 (LPA2)

+: LPA2 IC50 comprised between about 100 nM and 200 nM ++: LPA2 IC50 comprised between about 45 nM and 100 nm +++: LPA2 IC50 less than about 45 nM. Comparative Examples A-H

The comparative Examples A-H known in the art for their inhibitory activity on receptor LPA1, have been tested in the in vitro assay for the determination of activity also on LPA2 receptor as described above.

Table 3 Differently from the compounds of formula (I) of the present invention, the Comparative Examples A-H do not show a remarkable activity on the receptor LPA2. In fact, their inhibitory activity expressed as IC50 on LPA2 is comprised between 0.8 and 6 mM and therefore it is significantly lower in comparison with the inhibitory activity of the compound of the present invention.