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

Lysophosphatidic acid (LPA) is a phospholipid mediator concentrated in serum that acts as a potent extracellular signalling 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. <NPL>). 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 <NUM>:<NUM> oleoyl-LPA (<NUM>-acyl-<NUM>-hydroxy-sn-glycero3-phosphate), that is the most quantitatively abundant forms of LPA in human plasma with <NUM>:<NUM>-, <NUM>:<NUM>-, and <NUM>:<NUM>-LPA (see e.g.<NPL>). 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). Autotoxin (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. <NPL>).

ATX activity is the major source of plasma extracellular LPA but the source of tissue LPA that contributes to signalling 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 <NUM>-TM proteins that signal through at least two of the four Gα subunit families (Gα12/<NUM>, Gαq/<NUM>, Gαi/o and GαS). LPA receptors usually trigger response from multiple heterotrimeric G-proteins, resulting in diverse outcomes in a context and cell type dependent manner. Gα12/<NUM>-mediated LPA signalling regulates cell migration, invasion and cytoskeletal re-adjustments through activation of RHO pathway proteins. RAC activation downstream of Gαi/o-PI3K also regulates similar processes, but the most notable function of LPA-induced Gαi/o is mitogenic signalling through the RAF-MEK-MAPK cascade and survival signalling through the PI3K-AKT pathway. The LPA-coupled Gαq/<NUM> protein primarily regulates Ca2+ homeostasis through PLC and the second messengers IP3 and DAG. Lastly, GαS can activate adenylyl cyclase and increase cAMP concentration upon LPA stimulation (see e.g. <NPL>).

LPA, especially LPA1, LPA2 and LPA3, have been implicated in migration, invasion, metastasis, proliferation and survival and differ in their tissue distribution and downstream signalling pathways.

LPA1 is a <NUM>-kD protein that is widely expressed, albeit at different levels, in all human adult tissues examined and the importance of LPA1 signalling during development and adult life has been demonstrated through numerous approaches (see e.g. <NPL>). 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: Gαi/o, Gαq/<NUM>, and Gα12/<NUM>. 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. <NPL>).

LPA2 in humans is a <NUM>-kD protein and shares -<NUM>% amino acid sequence homology with LPA1 (see e.g. <NPL>). 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 signalling activity, LPA2 mostly activates the same pathways as triggered by LPA1 with some exceptions that regards its unique cross-talk behaviour. For example, LPA2 promotes cell migration through interactions with focal adhesion molecule TRIP6 (see e.g. <NPL>), and several PDZ proteins and zinc finger proteins are also reported to interact directly with the carboxyl-terminal tail of LPA2 (see e.g. <NPL>).

Human LPA3 is a <NUM>-kD protein and shares sequence homology with LPA1 (-<NUM>%) and LPA2 (-<NUM>%). 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 Gαi/o and Gαq/<NUM> (see e. Each LPA has multiple important regulatory functions throughout the body.

As LPA signalling has been strongly implicated in many disease states, great interest has been expressed in developing specific LPA inhibitors (see e.g.<NPL>). 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. <NPL>).

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. <NPL>). 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. <NPL> and <NPL>).

In vitro, LPA1 is known to induce the proliferation and differentiation of lung fibroblasts (see e.g. <NPL>), and to augment the fibroblast-mediated contraction of released collagen gels (see e.g. <NPL>). In human lung fibroblasts, the knockdown of LPA2 attenuated the LPA-induced expression of TGF-β1 and the differentiation of lung fibroblasts to myofibroblasts, resulting in the decreased expression of different profibrotic markers such as FN, α-SMA, and collagen, as well as decreased activation of extracellular regulated kinase <NUM>/<NUM>, Akt, Smad3, and p38 mitogen-activated protein kinase (see e.g. <NPL>). 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-β 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. <NPL>). In in vivo preclinical model, the oral administration of an LPA1 antagonist significantly reduced bleomycin-induced pulmonary fibrosis in mice (<NPL>; <NPL>), and the intraperitoneal injection of an LPA1/<NUM> antagonist ameliorated irradiation-induced lung fibrosis (see e.g.<NPL>). In a renal fibrosis model, LPA1 administration of an LPA1 antagonist suppressed renal interstitial fibrosis (see e.

Various compounds have been described in the literature as LPA1 or LPA2 antagonist.

<CIT> and <CIT> disclose cyclohexyl acid isoxazole azines as LPA1 antagonist, useful for the treatment of disorder or condition associated with dysregulation of lysophosphatidic acid receptor <NUM>.

<CIT> discloses isoxazole N-linked carbamoyl cyclohexyl acid as LPA1 antagonist for the treatment of disorder or condition associated with dysregulation of lysophosphatidic acid receptor <NUM>.

<CIT> 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 <NUM>.

<CIT> discloses carbamoyloxymethyl triazole cyclohexyl acids as LPA1 antagonist for the treatment of fibrosis including idiopathic pulmonary fibrosis.

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

<CIT> discloses phenyl isoxazole carbamate derivatives as LPA1 antagonist for the treatment of LPA mediated disorder, such as fibrosis.

discloses in "<NPL>, LPA2 antagonists. Key compounds were evaluated in vitro for inhibition of LPA2 mediated Erk activation and proliferation of HCT-<NUM> cells. These compounds could be used as tool compounds to evaluate the anticancer effects of blocking LPA2 signalling.

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

Despite the above cited prior art, there remains a potential for developing novel inhibitors of receptors LPA1 with a suitable BSEP (Bile Salt Export Pump inhibition) profile and good permeability 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 amido cyclohexane acid derivatives of general formula (I) of the present invention having antagonist activity on receptors LPA1 and at the same time a suitable BSEP profile and a good permeability which represent a solution to the aforementioned need.

In a first aspect the invention refers to a compound of formula (I)
<CHM>.

In a second aspect, the invention refers to pharmaceutical composition comprising a compound of formula (I) in a mixture with one or more pharmaceutically acceptable carrier or excipient.

In a third aspect, the invention refers to a compound of formula (I) for the use as a medicament.

In a further aspect, the invention refers to a compound of formula (I) for use in treating disease, disorder, or condition associated with dysregulation of lysophosphatidic acid receptor <NUM> (LPA1).

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

Unless otherwise provided, the term compound of formula (I) comprises in its meaning stereoisomer, tautomer or pharmaceutically acceptable salt or solvate.

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 Diastereomer s 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 (<NPL>)).

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 "<NUM>-membered heterocyclyl" refers to a mono satured or unsatured group containing one or more heteroatoms selected from N and O.

The term "(Cx-Cy) 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 <NUM> and y is <NUM>, for example, the term includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl and n-hexyl.

The term "(Cx-Cy)alkylene" wherein x and y are integers, refers to a Cx-Cyalkyl radical having in total two unsatisfied valences, such as a divalent methylene radical.

The expressions "(Cx-Cy) haloalkyl" wherein x and y are integers, refer to the above defined "Cx-Cyalkyl" 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 "(Cx-Cy) haloalkyl" groups may thus include halogenated, poly-halogenated and fully halogenated alkyl groups wherein all hydrogen atoms are replaced by halogen atoms, e.g. trifluoromethyl.

The term "(Cx-Cy) 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 <NUM> 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.

A bond pointing to a wavy or squiggly line, such as
<CHM>
as used in structural formulas herein, depicts the bond that is the point of attachment of the moiety or substituent to the core or backbone structure.

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

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 an antagonist property versus receptor LPA1.

Differently from similar compounds of the prior art, the compounds of formula (I) of the present invention are able to act as antagonist LPA1 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, the compounds of formula (I) of the invention have an activity as shown in Table <NUM>, wherein for each compound is reported the potency expressed as half maximal inhibitory concentration (IC<NUM>) on receptors.

As it can be appreciated, all the compounds of the present invention according to Table <NUM>, show a potency with respect to their inhibitory activity on receptor LPA1 below <NUM>, preferably below <NUM> and more preferably below <NUM>.

More advantagously, beyond the antagonist property versus receptor LPA1, the compounds of the present invention are also endowed with a suitable BSEP profile, that is relevant for the progression of any drug candidate.

The bile salt export pump (BSEP) is an efflux transporter located on the canalicular membrane of hepatic cells and is the primary transporter of bile acids from the hepatocyte to the biliary system. Together with other hepatic transporters of uptake and efflux, it is involved in the homeostasis of bile salts.

In the last decade, BSEP inhibition has emerged as an important mechanism that may contribute to the initiation of human drug-induced liver injury and therefore it is important to consider BSEP inhibition alongside when considering the risk of possible acute drug-induced liver failure.

BSEP inhibition was evaluated using human hepatocytes cultured between two layer of collagen (sandwich configuration). In this culture condition, hepatocytes express relevant transporters including BSEP and retain the bile canalicular structure. An inhibition of the biliary clearance of Taurocolic Acid (TCA), a known BSEP substrate, was used to assess BSEP interaction.

The compounds of formula (I) of the present invention are characterized by an in vitro BSEP inhibition at <NUM> ≤ <NUM> % that can be considared suitable and acceptable from a safety point of view, as shown in Table <NUM>.

Even more advantageously, the compounds of formula (I) of the present invention are also endowed with a good permeability profile that, in its turn, can ensure a suitable bioavailability for an oral administration. The permeability was assessed in human Caco <NUM> cell line, an in vitro model that mimic human gastrointestinal barrier and so useful to predic oral absorption. A passive permeability value ≥ <NUM>/sec is considered suitable for an oral administration, as shown in Table <NUM>.

Thus, in one aspect the present invention relates to a compound of general formula (I) as LPA1 antagonist
<CHM>.

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

In a preferred embodiment, the invention refers to at least one of the compounds listed in the Table <NUM> below and pharmaceutical acceptable salts thereof.

In one preferred embodiment, the invention refers to a compound of formula (I), wherein A is
<CHM>
and X is N, represented by the formula Ia
<CHM>
wherein.

In one preferred embodiment, the invention refers to compound of formula (Ia), wherein R<NUM> is selected from the group consisting of aryl, (C<NUM>-C<NUM>)cycloalkyl, heterocycloalkyl, and heteroaryl, wherein any of such aryl and heteroaryl is optionally substituted by one or more groups selected from (C<NUM>-C<NUM>)alkyl, halo, (C<NUM>-C<NUM>)haloalkyl, CN;.

In a still preferred embodiment, the invention refers to at least one of the compounds listed in the Table <NUM> below and pharmaceutical acceptable salts thereof.

In a further preferred embodiment, the invention refers to a compound of formula (I),
<CHM>.

It has been surprisingly found that the above indicated compounds are particularly effective as antagonists of LPA1 receptor, as e.g. indicated on Table <NUM> of the herein below experimental part.

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<NUM>) on LPA1 lesser than <NUM>.

Preferably, the compounds of the present invention have an IC<NUM> on LPA1 lesser or equal than <NUM>.

More preferably, the compounds of the present invention have an IC<NUM> on LPA1 lesser or equal than <NUM>.

The compounds of the present invention are also characterized by a BSEP inhibition at <NUM> ≤ <NUM> %.

The compounds of the invention are also characterized by a passive permeability value ≥ <NUM>/sec.

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

In a preferred embodiment, the invention refers to a compound of formula (I) 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 <NUM> (LPA1).

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

The terms "fibrosis" or "fibrosis 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 for use in 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 for use in the treatment of idiopathic pulmonary fibrosis (IPF).

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) 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) 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.

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, intraternally 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 compounds of the present invention can be prepared in a number of ways known to one skilled in the art of organic synthesis. It will be understood by those skilled in the art of organic synthesis that the functionality present on the molecule should be consistent with the transformation proposed. This will sometimes require a modification of the order of synthetic steps in order to obtain a desired compound of the invention. 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 <NUM> describes the synthesis of isoxazole amido cyclohexane acid derivatives of formula (XI). A <NUM>-nitrobenzoic acid or a <NUM>-nitro picolinic acid (II) is converted to the corresponding acid chloride using a chlorinating agent such as SOCl<NUM> or Oxalyl chloride/catalytic DMF. This acid chloride is then reacted with a suitable β-enamino-ester (III) followed by condensation with hydroxylamine to provide isoxazole (IV). Deprotection of the ester and subsequent Curtius rearrangement in the presence of the commercially available alcohol (VI) provide the isoxazole carbamate (VII). Reduction of the nitro group under suitable conditions such as iron (Fe) in acidic conditions (ex. HCl) leads to the amino intermediate (VIII). Final compound (XI) can be obtained through amide coupling with a cyclohexane dicarboxylic acid mono ester of formula (IX) in the presence of an appropriate coupling reagent (e.g. HATU) followed by ester deprotection. Alternatively, intermediate (VIII) can be reacted with the commercially available anhydride (X) to provide directly final compound (XI).

Scheme <NUM> describes an alternative synthetic route to isoxazole amido cyclohexane acid derivatives of formula (XI). A <NUM>-halo benzoic acid or a <NUM>-halo picolinic acid (XII) is converted to the corresponding isoxazole carbamate (XIII) by the same synthetic sequence previously outlined in Scheme <NUM>. Reaction of isoxazole (XIII) with benzophenone imine (XIV) under Buchwald reaction conditions (e.g. <NPL>) followed by cleavage of the resulting imines (XV) under well-known procedures (e.g. hydroxylamine hydrochloride) provides intermediate (VIII). Final compound (XI) is then obtained by following the synthetic sequence previously outlined in Scheme <NUM>.

Scheme <NUM> describes another alternative synthetic route to isoxazole amido cyclohexane acid derivative (XI). Reaction of intermediate (XVI) with <NUM>-methoxybenzylamine leads to PMB-protected intermediate (XVII). Subsequent deprotection under well-known procedures such as under strongly acidic conditions provides intermediate (XVIII) which is then reacted with the commercially available anhydride (X) to provide compound (XIX). Tert-butylation in the presence of a suitable reagent, such as N,N-Dimethylformamide di-tert-butyl acetal, provides intermediate (XX) which undergoes basic hydrolysis of the methyl ester followed by Curtius rearrangement to give compound (XXI). Final hydrolysis of the tert-butyl ester under acidic conditions leads to the final compound (XI).

In another embodiment of the present invention, wherein R3 is not H, compound (XXIV) may be obtained according to Scheme <NUM>. Reaction of Intermediate (VIII) with a suitable aldehyde (XXII) under reductive amination conditions provides compound (XXIII) which undergoes the same synthetic sequence previously outlined in Scheme <NUM> to provide final compound (XXIV).

Scheme <NUM> describes the synthesis of pyrazole amido cyclohexane acid derivatives of formula (XXX). An appropriately protected halo-pyrazole ester (XXV) undergoes borylation (e.g. using pinacol diboronate in the presence of a suitable palladium catalyst such as [<NUM>,<NUM>'-Bis(diphenylphosphino)ferrocene]dichloropalladium(II)) to give boronate (XXVI) which is then subjected to Suzuki-Miyaura coupling with an appropriate <NUM>-nitro phenyl/pyridine halide (XXVII) to provide the corresponding <NUM>-nitro phenyl/pyridyl pyrazole (XXVIII). After deprotection, the obtained carboxylic acid (XXIX) is carried forward to the final compound (XXX) following the same synthetic route described in Scheme <NUM>.

Scheme <NUM> describes an alternative synthetic route to obtain pyrazole amido cyclohexane acid derivative (XXX). A <NUM>-nitro phenyl/pyridine halide (XXVII) undergoes borylation to give boronate (XXXI) which is then subjected to Suzuki-Miyaura coupling with the halo-pyrazole (XXXII) to afford halo-pyrazole amine (XXXIII). Subsequent alkoxycarbonylation (e.g. using N,N'-Disuccinimidyl Carbonate) in the presence of the commercially available alcohol (VI) provides carbamate (XXXIV). Final compound (XXX) is then obtained by following the synthetic sequence previously outlined in Scheme <NUM>.

Scheme <NUM> describes the synthesis of triazole amido cyclohexane acid derivative (XL). A <NUM>-nitro phenyl/pyridine halide (XXVII) undergoes Sonogashira coupling with propargyl alcohol (XXXV) in the presence of a suitable palladium catalyst such as Bis(triphenylphosphine)palladium(II) dichloride to give the corresponding nitrophenyl/pyridinyl propargyl alcohol (XXXVI). Subsequent reaction with alkyl azide (XXXVII) with an appropriate catalyst provides the corresponding triazole alcohol (XXXVIII) which is then reacted with an oxidizing reagent (e.g. Potassium permanganate) to afford triazole carboxylic acid (XXXIX). Final compound (XL) is then obtained by the same synthetic sequence described in Scheme <NUM>.

Scheme <NUM> describes an alternative synthetic route to obtain triazole amido cyclohexane acid derivative (XL). Reaction of <NUM>-amino phenyl/pyridine halide (XLI) with the commercially available anhydride (X) and subsequent carboxylic acid protection provide intermediate (XLII). Subsequent Sonogashira coupling with propargyl alcohol in the presence of a suitable palladium catalyst such as Bis(triphenylphosphine)palladium(II) dichloride gives the corresponding propargyl (XXXV) alcohol intermediate (XI,III). Reaction of the latter with alkyl azide (XXXVII) with an appropriate catalyst followed by oxidation with a suitable oxidizing agent, such as Potassium permanganate, provides triazole carboxylic acid (XI,IV). Curtius rearrangement in the presence of the commercially available alcohol (VI) and final carboxylic acid deprotection provide the triazole amido cyclohexane acid derivative (XL).

Alternatively, compound (XL) may be obtained according to Scheme <NUM>. A nitro phenyl/pyridine-propynol (XXXVI), protected as tert-Butyldimethylsilyl ether (XLV), undergoes cycloaddition with Trimethylsilyl azide to afford the triazole (XLVI). Alkylation in the presence of a suitable base such as K<NUM>CO<NUM> provides the R4-substituted triazole (XLVII). Deprotection and subsequent oxidation of the alcohol provide the corresponding triazole carboxylic acid (XXXIX) which undergoes the same synthetic sequence previously outlined in Scheme <NUM> to provide final compound (XL).

In another embodiment of the present invention, wherein R4 = CH<NUM>, compound (LI) may be obtained according to Scheme <NUM>. Trimethylsilyldiazomethane can be used for the cycloaddition to the nitro phenyl/pyridine-propynol (XXXVI) to afford, after desilylation, N-methyl triazole (XLIX). Oxidation of the alcohol provides the corresponding triazole carboxylic acid (L) which undergoes the same synthetic sequence previously outlined in Scheme <NUM> to provide final compound (LI).

Scheme <NUM> describes the synthesis of tiophene amido cyclohexane acid derivatives of formula (LV). A <NUM>-nitro phenyl/pyridine boronate (XXXI) undergoes Suzuki-Miyaura coupling with a halo-thiophene carboxylic acid ester (LII) to afford a phenyl/pyridine thiophene carboxylic acid ester (LIII). Deprotection of the latter leads to carboxylic acid (LIV) which undergoes the same synthetic sequence previously outlined in Scheme <NUM> to provide final compound (LV).

Scheme <NUM> describes an alternative synthetic route to obtain tiophene amido cyclohexane acid derivatives of formula (LV). Thiophene carboxylic acid (LVI) undergoes Curtius rearrangement in the presence of the commercially available alcohol (VI) to afford the corresponding carbamate (LVII). Thiophene bromination with a suitable reagent such as N-Bromosuccinimide provides intermediate (LVIII) which is reacted with a <NUM>-amino phenyl/pyridine organotin compound (LIX) under Stille cross coupling conditions (e.g. <NPL>) to afford compound (LX). Final compound (LV) is then obtained by following the same synthetic sequence described in Scheme <NUM>.

Scheme <NUM> describes the synthesis of thiazole amido cyclohexane acid derivatives of formula (LXIV). Bromo isothiazole carboxylic acid (LXI) undergoes Curtius rearrangement in the presence of a commercially available alcohol (VI) to afford the corresponding carbamate (LXII). Subsequent reaction with a <NUM>-amino phenyl/pyridine organotin compound (LIX) under Stille cross coupling conditions (e.g. <NPL>) affords compound (LXIII). Final compound (LXIV) is then obtained by following the same synthetic sequence described in Scheme <NUM>.

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.

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.

<NUM>H-NMR spectra were performed on a Varian MR-<NUM> spectrometer operating at <NUM> (proton frequency), equipped with: a self-shielded Z-gradient coil <NUM> <NUM>/nX broadband probe head for reverse detection, deuterium digital lock channel unit, quadrature digital detection unit with transmitter offset frequency shift, or on AgilentVNMRS-<NUM> or on a Bruker Avance <NUM> spectrometers or on a bruker Fourier <NUM>. Chemical shift are reported as <NUM> 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 singlet, nd=not determined).

LC/MS retention times are estimated to be affected by an experimental error of +<NUM>. LCMS may be recorded under the following conditions: diode array DAD chromatographic traces, mass chromatograms and mass spectra may be taken on UPLC/PDA/MS AcquityTM system coupled with Micromass ZQTM or Waters SQD single quadrupole mass spectrometer operated in positive and/or negative electron spray ES ionization mode and/or Fractionlynx system used in analytical mode coupled with ZQTM single quadrupole operated in positive and/or negative ES ionisation mode or on a Shimadzu LCMS-<NUM> Single Quadrupole Liquid Chromatograph Mass Spectrometer and LCMS spectra were measured on Dionex UHPLC Ultimate <NUM> with DAD detector/Thermo Scientific MSQ Plus.

Quality Control methods used operated under low pH conditions or under high pH conditions:.

Chiral resolutions were performed using both Semipreparative HPLC (Agilent <NUM> system and Waters <NUM> system) and SFC (SFC preparative system from Jasco) technologies.

Where the preparation of starting materials is not described, these are commercially available, known in the literature, or readily obtainable by those skilled in the art using standard procedures.

Flash chromatography is carried out using an Isolera MPLC system (manufactured by Biotage) using pre-packed silica gel or reverse-phase cartridges (supplied by Biotage).

Many of the compounds described in the following Examples have been prepared from stereochemically pure starting materials, for example <NUM>% ee.

The stereochemistry of the compounds in the Examples, where indicated, has been assigned on the assumption that absolute configuration at resolved stereogenic centers of staring materials is maintained throughout any subsequent reaction conditions.

In the procedures that follow, after each starting material, reference to a compound number is sometimes provided. This is provided merely for assistance to the skilled chemist. The starting material may not necessarily have been prepared from the batch referred to.

When reference is made to the use of a "similar" or "analogous" procedure, as will be appreciated by those skilled in the art, such a procedure may involve minor variations, for example reaction temperature, reagent/solvent amount, reaction time, work-up conditions or chromatographic purification conditions.

Oxalyl dichloride (<NUM>, <NUM> mmol) and dry DMF (<NUM>, <NUM> mmol) were added at <NUM> to a suspension of <NUM>-bromo-<NUM>-methyl picolinic acid (<NUM>, <NUM> mmol) in dry DCM (<NUM>). The mixture was stirred at r. for <NUM> and the solvent was removed under reduced pressure to provide the title compound (<NUM>, crude) as a brown solid. <NUM>H NMR (<NUM>, DMSO-d<NUM>) δ ppm <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>).

To a solution of methyl (E)-<NUM>-(methylamino)but-<NUM>-enoate (<NUM>, <NUM> mmol), prepared according to the procedure reported in <NPL>, in dry THF (<NUM>), Pyridine (<NUM>, <NUM> mmol) was added dropwise. The reaction was cooled in an ice bath and a solution of <NUM>-bromo-<NUM>-methylpicolinoyl chloride (Intermediate A1. <NUM>, <NUM>, <NUM> mmol)) in dry THF (<NUM>) was slowly added. The reaction was warmed up to room temperature and stirred overnight. H<NUM>O was added (<NUM>) and the solution was extracted with EtOAc for <NUM> times. The combined organic layer was further washed with water and brine, dried over Na<NUM>SO<NUM> and evaporated under reduced pressure to provide the title compound (<NUM>, crude) as a brown oil that was used in the next step without further purification. LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>).

(Intermediate A1. <NUM>, <NUM>, crude) was dissolved in Acetic acid (<NUM>) and hydroxylamine hydrochloride (<NUM>, <NUM> mmol) was added and the mixture was heated at <NUM> for <NUM>. The mixture was evaporated under vacuum, NaHCOs saturated aqueous solution (<NUM>) was added and the mixture was extracted with EtOAc for <NUM> times. The combined organic layer was further washed with water and brine, dried over Na<NUM>SO<NUM>, and evaporated under reduced pressure. The crude was purified by flash chromatography eluting with a gradient of cyclohexane/EtOAc from <NUM>/<NUM> to <NUM>/<NUM> to provide the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a white solid.

LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>)
<NUM>H NMR (<NUM>, Chloroform-d) δ ppm <NUM> (d, J=<NUM>, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>)
The Intermediates in the following table were prepared from reagents reported below by using methods analogous to Intermediate A1.

A mixture of methyl <NUM>-bromo-<NUM>-methyl-<NUM>H-pyrazole-<NUM>-carboxylate (<NUM>, <NUM> mmol), potassium acetate (<NUM>, <NUM> mmol) and Bis(pinacolato)diboron (<NUM>, <NUM> mmol) in <NUM>,<NUM>-Dioxane (<NUM>) was degassed under N<NUM> for <NUM> minutes, then Pd(dppf)Cl<NUM> (<NUM>, <NUM> mmol) was added. The mixture was stirred at <NUM> overnight. Water was added and the mixture was extracted with EtOAc for <NUM> times, collected organic fractions were dried over Na<NUM>SO<NUM>, filtered and evaporated to give the title compound (<NUM>, crude) as brown oil which was used in the next step without further purification. LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>).

To a solution of methyl <NUM>-methyl-<NUM>-(<NUM>,<NUM>,<NUM>,<NUM>-tetramethyl-<NUM>,<NUM>,<NUM>-dioxaborolan-<NUM>-yl)-<NUM>H-pyrazole-<NUM>-carboxylate (Intermediate B1. <NUM>, <NUM>, <NUM> mmol) in <NUM>,<NUM>-Dioxane (<NUM>) <NUM>-bromo-<NUM>-methyl-<NUM>-nitropyridine (<NUM>, <NUM> mmol), K<NUM>HPO<NUM> (<NUM>, <NUM> mmol), Water (<NUM>) and X-Phos Pd G2 (<NUM>, <NUM> mmol) were added under nitrogen. The mixture was stirred at <NUM> overnight. Water was added and the mixture was extracted with EtOAc for <NUM> times, collected organic phases were dried over Na<NUM>SO<NUM>, filtered and evaporated under reduced pressure. The residue was purified by flash chromatography eluting with a gradient of EtOAc in CyHex from <NUM>% to <NUM>% to afford the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a pink solid.

A mixture of ethyl <NUM>-bromothiophene-<NUM>-carboxylate (<NUM>, <NUM> mmol), (<NUM>-nitrophenyl)boronic acid (<NUM>, <NUM> mmol), Na<NUM>CO<NUM> (<NUM>, <NUM> mmol) in <NUM>,<NUM>-dimethoxyethane (<NUM>) and Water (<NUM>) was degassed by applying alternatively vacuum and nitrogen. Pd(dppf)Cl<NUM> (<NUM>, <NUM> mmol) was then added and the mixture was heated at <NUM> for <NUM>. The mixture was then diluted with EtOAc, washed with water and brine, dried over Na<NUM>SO<NUM> and evaporated under reduced pressure. The residue was purified by flash chromatography eluting with a gradient of EtOAc in CyHex from <NUM>% to <NUM>% to afford the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a white solid.

A stirred suspension in anhydrous toluene (<NUM>) of methyl <NUM>-(<NUM>-bromo-<NUM>-methylpyridin-<NUM>-yl)-<NUM>-methyl-<NUM>,<NUM>-oxazole-<NUM>-carboxylate (Intermediate A1, <NUM>, <NUM> mmol), <NUM>-methoxybenzylamine (<NUM>, <NUM> mmol) and cesium carbonate (<NUM>, <NUM> mmol) was purged with argon for <NUM>, followed by addition of Pd<NUM>(dba)<NUM>CHCl<NUM> (<NUM>, <NUM> mmol) and XantPhos (<NUM>, <NUM> mmol). The reaction was stirred overnight at <NUM>. The mixture was filtered and the solvent was evaporated to obtain an oil which was dissolved in DCM and precipitated with CyHex. The crude was purified by flash chromatography using a gradient of EtOAc in n-hexane from <NUM>% to <NUM>% affording the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a yellow solid.

<NUM> HCl in <NUM>,<NUM>-Dioxane (<NUM>, <NUM> mmol ) was added to methyl <NUM>-(<NUM>-{[(<NUM>-methoxyphenyl)methyl]amino}-<NUM>-methylpyridin-<NUM>-yl)-<NUM>-methyl-<NUM>,<NUM>-oxazole-<NUM>-carboxylate (Intermediate D1. <NUM>) (<NUM>, <NUM> mmol) in <NUM>,<NUM>-Dioxane (<NUM>). The reaction was stirred overnight at <NUM>, the solvent was evaporated, then DCM and water were added and the mixture was basified with NH<NUM>OH to pH <NUM>. The layers were separated and water phase was washed with DCM (3x <NUM>). Combined organic layers were washed with brine (<NUM>) and dried over sodium sulfate to give <NUM> of crude. The product was triturated in Et<NUM>O (<NUM>) to provide the title compound as a yellow solid (<NUM>, <NUM> mmol, <NUM>% yield).

(-) Trans-<NUM>,<NUM>-Cyclohexanedicarboxylic anhydride (<NUM>, <NUM> mmol) was added to a solution of methyl <NUM>-(<NUM>-amino-<NUM>-methylpyridin-<NUM>-yl)-<NUM>-methyl-<NUM>,<NUM>-oxazole-<NUM>-carboxylate (Intermediate D1. <NUM>) (<NUM>, <NUM> mmol) in DMF (<NUM>). The mixture was stirred overnight at <NUM>. The solution was diluted with water (<NUM>) and the product was extracted with EtOAc (3x250 mL). Combined organic layers were washed with <NUM>% KHSO<NUM> (3x <NUM>), dried over sodium sulphate and evaporated to give the title compound as a yellow foam (<NUM>, <NUM> mmol, <NUM>% yield).

(<NUM>,<NUM>)-<NUM>-({<NUM>-[<NUM>-(methoxycarbonyl)-<NUM>-methyl-<NUM>,<NUM>-oxazol-<NUM>-yl]-<NUM>-methylpyridin-<NUM>-yl}carbamoyl)cyclohexane-<NUM>-carboxylic acid (Intermediate D1. <NUM>) (<NUM>, <NUM> mmol) was dissolved in anhydrous toluene (<NUM>) under argon. The mixture was heated to reflux. N,N-dimethylformamide di-tert-butyl acetal (<NUM>, <NUM> mmol) was added dropwise by syringe pump over <NUM> to the refluxing mixture. Additional portion of N,N-dimethylformamide di-tert-butyl acetal (<NUM>, <NUM> mmol) was added dropwise by syringe pump over <NUM> to the refluxing mixture. The reaction was cooled down to room temperature, then was diluted with EtOAc (<NUM>) and the organic layer was washed with sodium bicarbonate sat. solution (3x <NUM>), water (<NUM>) and brine (<NUM>). The organic layer was dried over sodium sulfate and evaporated to obtain <NUM> of crude as an oil. Purification by flash chromatography using a gradient of EtOAc in hexane from <NUM> to <NUM>% gave the title compound as a yellow foam (<NUM> <NUM> mmol. <NUM>% yield).

Methyl <NUM>-(<NUM>-bromo-<NUM>-methylpyridin-<NUM>-yl)-<NUM>-methylisoxazole-<NUM>-carboxylate (Intermediate A1, <NUM>, <NUM> mmol) was dissolved in THF (<NUM>) and lithium hydroxide <NUM> solution in H<NUM>O (<NUM>, <NUM> mmol) was added and the mixture was stirred at <NUM> for <NUM>. The mixture was acidified to neutral pH with HCl 1N in H<NUM>O and extracted with EtOAc for <NUM> times. The combined organic layer was further washed with water and brine, dried over Na<NUM>SO<NUM> and evaporated under reduced pressure to afford the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a pale yellow solid.

The Intermediates in the following table were prepared from reagents reported below by using methods analogous to Intermediate E1.

To a solution of <NUM>-bromo-<NUM>-methyl-<NUM>-nitropyridine (<NUM>, <NUM> mmol) in dry MeCN (<NUM>), <NUM>-propyn-<NUM>-ol (<NUM>, <NUM> mmol) and triethylamine (<NUM>, <NUM> mmol) were added, followed by copper iodide (<NUM>, <NUM> mmol) and PdCl<NUM>(PPh<NUM>)<NUM> (<NUM>, <NUM> mmol) at <NUM>. The reaction mixture was stirred at r. for <NUM>, then the salts were filtered off through a celite pad and the filtrate was concentrated under reduced pressure. The crude was purified by flash chromatography using a gradient of EtOAc in cyclohexane from <NUM>% to <NUM>% affording the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a brown solid.

To a solution of <NUM>-(<NUM>-methyl-<NUM>-nitropyridin-<NUM>-yl)prop-<NUM>-yn-<NUM>-ol (Intermediate F1. <NUM>, <NUM>, <NUM> mmol) in dry <NUM>,<NUM>-Dioxane (<NUM>), trimethylsilylmethyl azide (<NUM>, <NUM> mmol) was added. Three nitrogen/vacuum cycles were applied, then Cp*RuCl(PPh<NUM>)<NUM> (<NUM>, <NUM> mmol) was added. The mixture was stirred at <NUM> for <NUM> , then concentrated under reduced pressure to afford a crude mixture that was purified by flash chromatography using a gradient of EtOAc in cyclohexane from <NUM>% to <NUM>% affording the title compound (<NUM>, <NUM> mmol, <NUM>% yield). LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>).

To a solution of (<NUM>-(<NUM>-methyl-<NUM>-nitropyridin-<NUM>-yl)-<NUM>-((trimethylsilyl)methyl)-<NUM>H-<NUM>,<NUM>,<NUM>-triazol-<NUM>-yl)methanol (Intermediate F1. <NUM>, <NUM>, <NUM> mmol) in dry THF (<NUM>), TBAF <NUM> in THF (<NUM>, <NUM> mmol) was added at <NUM>. The reaction mixture was stirred for <NUM>, then solid NaHCOs was added and the reaction was vigorously stirred at RT for <NUM>. The solid was filtered off and the filtrate concentrated under reduced pressure. The crude was purified by flash column chromatography using a gradient of EtOAc in cyclohexane from <NUM>% to <NUM>% affording the title compound (<NUM>, <NUM> mmol, <NUM> % yield) as a pale orange solid. LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>).

To a solution of (<NUM>-methyl-<NUM>-(<NUM>-methyl-<NUM>-nitropyridin-<NUM>-yl)-<NUM>H-<NUM>,<NUM>,<NUM>-triazol-<NUM>-yl)methanol (Intermediate F1. <NUM>, <NUM>, <NUM> mmol) in MeCN (<NUM>) and Water (<NUM>), KMnO<NUM> (<NUM>, <NUM> mmol) was added and the resulting purple solution was stirred at r. HCl <NUM> was then added up to pH < <NUM> and the solvent was removed under reduced pressure. The crude was purified by flash chromatography using a gradient of MeOH in DCM from <NUM>% to <NUM>% affording the title compound (<NUM>, <NUM> mmol, <NUM> % yield).

The Intermediate in the following table was prepared from reagents reported below by using methods analogous to Intermediate F1.

<NUM>-(<NUM>-methyl-<NUM>-nitropyridin-<NUM>-yl)prop-<NUM>-yn-<NUM>-ol (Intermediate F1. <NUM>, <NUM>, <NUM> mmol) and imidazole (<NUM>, <NUM> mmol) were dissolved in THF (<NUM>) and DMF (<NUM>), then tert-butyl-chloro-dimethylsilane (<NUM>, <NUM> mmol) was added under nitrogen atmosphere. The reaction mixture was stirred overnight at <NUM>. Water was added and the organic phase was removed under reduced pressure. The residue was extracted with EtOAc, washed sequentially with a saturated NaHCOs solution and brine. Organic phase was evaporated and the crude was purified by flash chromatography using a gradient of EtOAc in cyclohexane from <NUM>% to <NUM>% to give the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a pale-yellow oil. LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>).

To a solution of <NUM>-(<NUM>-((tert-butyldimethylsilyl)oxy)prop-<NUM>-yn-<NUM>-yl)-<NUM>-methyl-<NUM>-nitropyridine (Intermediate G1. <NUM>, <NUM>, <NUM> mmol) in DMF (<NUM>), trimethylsilyl azide (<NUM>, <NUM> mmol) was added. The reaction was set under N<NUM> and heated at <NUM> for <NUM>. The solvent was removed under reduced pressure and the crude was purified via reverse phase flash chromatography using a gradient of MeCN (<NUM>% HCOOH) in acidic water (<NUM>% HCOOH) from <NUM> to <NUM>% to afford the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a yellow-orange solid. LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>).

A solution of <NUM>-(<NUM>-(((tert-butyldimethylsilyl)oxy)methyl)-<NUM>H-<NUM>,<NUM>,<NUM>-triazol-<NUM>-yl)-<NUM>-methyl-<NUM>-nitropyridine (Intermediate G1. <NUM>, <NUM>, <NUM> mmol) in MeCN (<NUM>) was cooled at <NUM> and potassium carbonate (<NUM>, <NUM> mmol) was added, followed by iodoethane (<NUM>µL, <NUM> mmol). The reaction was stirred at r. for <NUM> hours, then the solvent was removed under reduced pressure and the residue was taken up with DCM and water. The water phase was extracted trice with DCM. The combined organic phase was filtered and the solvent was removed under reduced pressure. The crude was then purified by flash column chromatography using a gradient of EtOAc in cyclohexane from <NUM> to <NUM>% to afford the title compound (<NUM>, <NUM> mmol, <NUM> % yield) as a white solid. LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>).

To a solution of <NUM>-(<NUM>-(((tert-butyldimethylsilyl)oxy)methyl)-<NUM>-ethyl-<NUM>H-<NUM>,<NUM>,<NUM>-triazol-<NUM>-yl)-<NUM>-methyl-<NUM>-nitropyridine (Intermediate G1. <NUM>, <NUM>, <NUM> mmol) in dry THF (<NUM>), tetrabutylammonium fluoride (<NUM>, <NUM> mmol) <NUM> in THF was added at <NUM>. The reaction mixture was stirred for <NUM>, then solid NaHCOs was added and the mixture was vigorously stirred at r. The solid was filtered off and the filtrate was concentrated under reduced pressure. The crude was purified by flash column chromatography using a gradient of EtOAc in cyclohexane from <NUM>% to <NUM>% to afford the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a white solid.

To a solution of (<NUM>-ethyl-<NUM>-(<NUM>-methyl-<NUM>-nitropyridin-<NUM>-yl)-<NUM>H-<NUM>,<NUM>,<NUM>-triazol-<NUM>-yl)methanol (Intermediate G1. <NUM>, <NUM>, <NUM> mmol,) in MeCN (<NUM>) and Water (<NUM>), KMnO<NUM> (<NUM>, <NUM> mmol) was added and the resulting purple solution was stirred at RT overnight. HCl <NUM> was then added up to pH < <NUM> and the solvent was removed under reduced pressure. The crude was purified by flash chromatography using a gradient of MeOH in DCM from <NUM>% to <NUM>% affording the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a white solid.

To a solution of <NUM>-bromo-<NUM>-fluoropyridin-<NUM>-amine (<NUM>, <NUM> mmol) in MeCN (<NUM>), (-)-trans-<NUM>,<NUM>-Cyclohexanedicarboxylic anhydride (<NUM>, <NUM> mmol) was added. The mixture was stirred at <NUM> for <NUM>. The solvent was removed under vacuum, then the crude was diluted with sat aq NH<NUM>Cl and extracted with EtOAc. Combined organic layers were evaporated under reduced pressure to provide the title compound (<NUM> mmol, crude) that was used in the next step without further purification.

(<NUM>,<NUM>)-<NUM>-((<NUM>-bromo-<NUM>-fluoropyridin-<NUM>-yl)carbamoyl)cyclohexane-<NUM>-carboxylic acid (Intermediate H1. <NUM>, <NUM> mmol, crude) was added to a solution of p-Toluenesulfonic Acid Monohydrate (<NUM>, <NUM> mmol) in MeOH (<NUM>). The mixture was stirred at <NUM> for <NUM>. The solvent was removed under vacuum, then the crude was diluted with sat. NH<NUM>Cl and extracted with EtOAc. The organic layer was evaporated under reduced pressure and the crude was purified by flash chromatography using a gradient of EtOAc in cyclohexane from <NUM>% to <NUM>% affording the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as an orange oil. LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>).

To a solution of methyl (<NUM>,<NUM>)-<NUM>-((<NUM>-bromo-<NUM>-fluoropyridin-<NUM>-yl)carbamoyl)cyclohexane-<NUM>-carboxylate (Intermediate H1. <NUM>, <NUM>, <NUM> mmol) in dry MeCN (<NUM>), <NUM>-propyn-<NUM>-ol (<NUM>, <NUM> mmol) and triethylamine (<NUM>, <NUM> mmol) were added, followed by copper iodide (<NUM>, <NUM> mmol) and PdCl<NUM>(PPh<NUM>)<NUM> (<NUM>, <NUM> mmol) at <NUM>. The reaction mixture was stirred at RT for <NUM>, then the salts were filtered off through a celite pad and the filtrate was concentrated under reduced pressure. The crude was purified by flash chromatography using a gradient of EtOAc in cyclohexane from <NUM>% to <NUM>% affording the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a brown solid.

To a solution of methyl (<NUM>,<NUM>)-<NUM>-((<NUM>-fluoro-<NUM>-(<NUM>-hydroxyprop-<NUM>-yn-<NUM>-yl)pyridin-<NUM>-yl)carbamoyl)cyclohexane-<NUM>-carboxylate (Intermediate H1. <NUM>, <NUM>, <NUM> mmol) in dry <NUM>,<NUM>-Dioxane (<NUM>), trimethylsilylmethyl azide (<NUM>, <NUM> mmol) was added. Three nitrogen/vacuum cycles were applied, then Cp*RuCl(PPh<NUM>)<NUM> (<NUM>, <NUM> mmol) was added. The mixture was stirred at <NUM> for <NUM> and concentrated under reduced pressure to afford a crude mixture that was purified by flash chromatography using a gradient of EtOAc in cyclohexane from <NUM>% to <NUM>% affording the title compound (<NUM>, <NUM> mmol, <NUM>% yield). LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>).

To a solution of methyl (<NUM>,<NUM>)-<NUM>-((<NUM>-fluoro-<NUM>-(<NUM>-(hydroxymethyl)-<NUM>-((trimethylsilyl)methyl)-<NUM>H-<NUM>,<NUM>,<NUM>-triazol-<NUM>-yl)pyridin-<NUM>-yl)carbamoyl)cyclohexane-<NUM>-carboxylate (Intermediate H1. <NUM>, <NUM>, <NUM> mmol) in dry THF (<NUM>), TBAF <NUM> in THF (<NUM>, <NUM> mmol) was added at <NUM>. The reaction mixture was stirred for <NUM>, then solid NaHCOs was added and the reaction was vigorously stirred at r. The solid was filtered off and the filtrate was concentrated under reduced pressure affording the title compound (<NUM> mmol, crude) that was used in the next step without further purification. LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>).

To a solution of methyl (<NUM>,<NUM>)-<NUM>-((<NUM>-fluoro-<NUM>-(<NUM>-(hydroxymethyl)-<NUM>-methyl-<NUM>H-<NUM>,<NUM>,<NUM>-triazol-<NUM>-yl)pyridin-<NUM>-yl)carbamoyl)cyclohexane-<NUM>-carboxylate (Intermediate H1. <NUM>, <NUM> mmol) in MeCN (<NUM>) and Water (<NUM>), KMnO<NUM> (<NUM>, <NUM> mmol) was added and the resulting purple solution was stirred at r. HCl <NUM> was then added up to pH < <NUM> and the solvent was removed under reduced pressure. The crude was purified by reverse phase flash chromatography using a gradient of MeCN in water from <NUM>% to <NUM>% affording the title compound (<NUM>, <NUM> mmol, <NUM> % yield). LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>).

A mixture of <NUM>-bromo-<NUM>-methylpyrazol-<NUM>-amine (<NUM>, <NUM> mmol), (<NUM>-nitrophenyl)boronic acid (<NUM>, <NUM> mmol), Pd(dppf)Cl<NUM> (<NUM>, <NUM> mmol) and K<NUM>PO<NUM> (<NUM>, <NUM> mmol) was degassed. Water (<NUM>) and <NUM>,<NUM>-Dioxane (<NUM>) were added and the mixture was stirred at <NUM> for <NUM>. The mixture was cooled to r. and concentrated under reduced pressure. The residue was extracted with DCM, the solvent was evaporated and the residue was triturated with MeOH to give the title compound (<NUM>, <NUM> mmol, crude) as a light brown solid that was used in the next step without further purification. LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>).

A <NUM> three-necked flask equipped with stir bar, reflux condenser, thermometer and nitrogen/vacuum stopcock was charged with Intermediate E1 (<NUM>, <NUM> mmol) and (R)-<NUM>-(<NUM>-chlorophenyl)ethanol (<NUM>, <NUM> mmol). The system was closed and three cycles of vacuum/nitrogen back-filling were applied. Dry Toluene (<NUM>) was added, followed by triethylamine (<NUM>, <NUM> mmol) and DPPA (<NUM>, <NUM> mmol). The solution was slowly heated to <NUM> and stirred at reflux over <NUM>. The resulting slurry was cooled to r. , filtered over a phase separator, and the filtrate was concentrated under reduced pressure. The crude was purified by flash column chromatography eluting with a gradient of EtOAc in cyclohexane from <NUM>% to <NUM>% to afford the title compound (<NUM>, <NUM> mmol, <NUM> % yield) as a pale yellow oil.

LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>)
The Intermediates in the following table were prepared from reagents reported below by using methods analogous to Intermediate J1.

To a solution of (R)-<NUM>-(<NUM>-chlorophenyl)ethanol (<NUM>, <NUM> mmol) in DCM (<NUM>)/MeCN (<NUM>), N,N'-Disuccinimidyl Carbonate (<NUM>, <NUM> mmol) and DMAP (<NUM>, <NUM> mmol) were added and the mixture was stirred for <NUM> at <NUM>. The reaction was cooled to r. then <NUM>-methyl-<NUM>-(<NUM>-nitrophenyl)-<NUM>H-pyrazol-<NUM>-amine (Intermediate I1, <NUM>, <NUM> mmol) was added and the solution was stirred at <NUM> overnight. The reaction mixture was concentrated and partitioned between a citrate buffer solution (pH <NUM>) and EtOAc, the organic layer was washed with NaHCO<NUM> saturated solution, dried over a phase separator and the solvent was evaporated under reduced pressure. The residue was purified by flash chromatography eluting with a gradient of EtOAc in cyclohexane from <NUM>% to <NUM>% to afford the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a yellow oil.

A <NUM> tube equipped with a stir bar was charged with Intermediate J1 (<NUM>, <NUM> mmol), Cs<NUM>CO<NUM> (<NUM>, <NUM> mmol), Pd<NUM>(dba)<NUM> (<NUM>, <NUM> mmol) and Xantphos (<NUM>, <NUM> mmol). The tube was sealed and three cycles of vacuum/nitrogen back-filling were performed. <NUM>,<NUM>-Dioxane (<NUM>) and benzophenone imine (<NUM>, <NUM> mmol) were added and three cycles of vacuum/nitrogen back-filling were repeated. The tube was sealed and the mixture was heated at <NUM> for <NUM>. The mixture was filtered, the solid was washed with EtOAc (3x5 mL) and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography eluting with a gradient of EtOAc in cyclohexane from <NUM>% to <NUM>% to provide the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a yellow oil. LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>).

(R)-<NUM>-(<NUM>-chlorophenyl)ethyl (<NUM>-(<NUM>-((diphenylmethylene)amino)-<NUM>-methylpyridin-<NUM>-yl)-<NUM>-methylisoxazol-<NUM>-yl)carbamate (Intermediate L1. <NUM>, <NUM>, <NUM> mmol) was dissolved in MeOH (<NUM>), then sodium acetate (<NUM>, <NUM> mmol) and hydroxylamine hydrochloride (<NUM>, <NUM> mmol) were sequentially added. The mixture was stirred at RT overnight and then concentrated under reduced pressure to remove most of the MeOH, diluted with sat. NaHCOs (<NUM>) and extracted with DCM (3x15 mL). The collected organic fractions were washed with brine (<NUM>), dried over Na<NUM>SO<NUM>, filtered and concentrated under reduced pressure. The oily brown residue was purified by flash column chromatography eluting with a gradient of EtOAc in cyclohexane from <NUM>% to <NUM>% to afford the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a yellow oil.

LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>)
The Intermediates in the following table were prepared from reagents reported below by using methods analogous to Intermediate L1.

(R)-<NUM>-(<NUM>-chlorophenyl)ethyl (<NUM>-methyl-<NUM>-(<NUM>-methyl-<NUM>-nitropyridin-<NUM>-yl)-<NUM>H-pyrazol-<NUM>-yl)carbamate (Intermediate J2, <NUM>, <NUM> mmol) was dissolved in DCM (<NUM>), MeOH (<NUM>) and concentrated HCl (<NUM>, <NUM> mmol), then Fe° (<NUM>, <NUM> mmol) was added and the mixture was stirred at r. The solvent was concentrated under reduced pressure and the residue was basified with a 2N aqueous NaOH solution and extracted with EtOAc <NUM> times. Collected organic fractions were washed with brine, dried over Na<NUM>SO<NUM>, filtered and concentrated under reduced pressure to provide the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a yellow solid, which was used in the next step without further purification.

LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>)
The Intermediates in the following table were prepared from reagents reported below by using methods analogous to Intermediate M1.

To a solution of (R)-<NUM>-(<NUM>-chlorophenyl)ethyl thiophen-<NUM>-ylcarbamate (Intermediate J10, <NUM>, <NUM> mmol) in DCM (<NUM>), N-Bromosuccinimide (<NUM>, <NUM> mmol) was added dropwise and the mixture was stirred for <NUM> at reflux. The reaction was cooled to r. , diluted with DCM (<NUM>), washed with sat. K<NUM>CO<NUM> (<NUM> x <NUM>), brine (<NUM>), dried over Na<NUM>SO<NUM>, filtered, and concentrated under reduced pressure. The residue was purified by flash chromatography eluting with a gradient of DCM in cyclohexane from <NUM>% to <NUM>% to afford the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as an off-white solid.

A <NUM> microwave vial equipped with a stir bar was charged with <NUM>-tributylstannylpyridin-<NUM>-amine (<NUM>, <NUM> mmol) and (R)-<NUM>-(<NUM>-chlorophenyl)ethyl (<NUM>-bromothiophen-<NUM>-yl)carbamate (Intermediate N1. <NUM>, <NUM>, <NUM> mmol). The vial was sealed and three cycles of vacuum/nitrogen back-filling were applied. Toluene (<NUM>) and DMF (<NUM>) were added and the mixture was sparged with nitrogen over <NUM>. Pd(PPh<NUM>)<NUM> (<NUM>, <NUM> mmol) was added, the tube was sealed and the mixture was heated at <NUM> for <NUM>. The mixture was cooled at r. , filtered over Celite and concentrated under reduced pressure. The residue was dissolved in MeOH (<NUM>) and treated with 2N KF (<NUM>) over <NUM>. The mixture was concentrated under reduced pressure, diluted with brine (<NUM>) and extracted with EtOAc (<NUM>). The organic phase was concentrated under reduced pressure and the residue was purified by flash column chromatography using a gradient of EtOAc in Cyclohexane from <NUM>% to <NUM>% to afford the target product (R)-<NUM>-(<NUM>-chlorophenyl)ethyl (<NUM>-(<NUM>-aminopyridin-<NUM>-yl)thiophen-<NUM>-yl)carbamate (Intermediate N1, <NUM>, <NUM> mmol, <NUM>. % yield) as an amorphous light brown solid.

To a solution of <NUM>-methyl-<NUM>-tributylstannylpyridin-<NUM>-amine (<NUM>, <NUM> mmol) and <NUM> (R)-<NUM>-(<NUM>-chlorophenyl)ethyl (<NUM>-bromoisothiazol-<NUM>-yl)carbamate (Intermediate J27, <NUM>, <NUM> mmol) in Toluene (<NUM>) and DMF (<NUM>), Pd(PPh<NUM>)<NUM> (<NUM>, <NUM> mmol) was added and the tube was sealed and heated at <NUM> for <NUM>. The mixture was cooled at r. , filtered over Celite and concentrated under reduced pressure. The residue was dissolved in MeOH (<NUM>) and treated with 2N KF (<NUM>) over <NUM>. The mixture was concentrated under reduced pressure, diluted with brine (<NUM>) and extracted with EtOAc (<NUM>). The organic phase was concentrated under reduced pressure and the residue was purified by flash column chromatography using a gradient of EtOAc in Cyclohexane from <NUM>% to <NUM>% to afford the target product (Intermediate O1, <NUM>, <NUM> mmol, <NUM>% yield) as a yellow solid.

To a solution of (R)-<NUM>-(<NUM>-chloropyridin-<NUM>-yl)ethyl (<NUM>-(<NUM>-aminopyridin-<NUM>-yl)-<NUM>-methylisoxazol-<NUM>-yl)carbamate (Intermediate M9, <NUM>, <NUM> mmol) in THF (<NUM>) formaldehyde (<NUM>, <NUM> mmol), MeCN (<NUM>) and sodium methoxide (<NUM>, <NUM> mmol) were added (from clear to orange solution). The reaction mixture was stirred at <NUM> overnight (yellow solution), then sodium borohydride (<NUM>, <NUM> mmol) was added (orange solution and gas evolution). The mixture was stirred at <NUM> for <NUM> (yellow solution) The solvent was removed under reduced pressure, NaHCOs satd. was added and the mixture was extracted with EtOAc. The solution was dried over Na<NUM>SO<NUM> and concentrated under reduced pressure. The residue was purified by flash chromatography eluting with a gradient of EtOAc in cyclohexane from <NUM> to <NUM>% to afford the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a pale orange solid.

To a solution of (1R)-<NUM>-(<NUM>-chlorophenyl)ethyl N-[<NUM>-(<NUM>-aminophenyl)-<NUM>-methyl-<NUM>-pyrazol-<NUM>-yl]carbamate (Intermediate M3, <NUM>, <NUM> mmol) and (±)-Trans-<NUM>-methoxycarbonylcyclohexane-<NUM>-carboxylic acid (<NUM>, <NUM> mmol), in MeCN (<NUM>), <NUM>-methylimidazole (<NUM>, <NUM> mmol) was added, followed by HATU (<NUM>, <NUM> mmol) and the mixture was stirred at r. The mixture was concentrated under reduced pressure then water was added to the residue and the mixture was extracted with EtOAc. The solvent was dried over Na<NUM>SO<NUM> and concentrated under reduced pressure. The residue was purified by flash chromatography eluting with cyclohexane:EtOAc <NUM>: <NUM> to afford the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a white foam LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>)
The Intermediates in the following table were prepared from reagents reported below by using methods analogous to Intermediate Q1.

To a solution of (1R)-<NUM>-(<NUM>-chlorophenyl)ethyl N-(<NUM>-(<NUM>-aminophenyl)thiophen-<NUM>-yl)carbamate (Intermediate M4, <NUM>, <NUM> mmol) and (±)-Cis-<NUM>-methoxycarbonylcyclohexane-<NUM>-carboxylic acid (<NUM>, <NUM> mmol) in MeCN (<NUM>), <NUM>-methylimidazole (<NUM>, <NUM> mmol) was added, followed by HATU (<NUM>, <NUM> mmol) and the mixture was stirred at r. The mixture was concentrated under reduced pressure then water was added to the residue and the mixture was extracted with EtOAc. The solvent was dried over Na<NUM>SO<NUM> and concentrated under reduced pressure. The residue was purified by flash chromatography using a gradient of EtOAc in cyclohexane from <NUM>% to <NUM>% to afford the title compound (<NUM>, <NUM> mmol, <NUM>% yield) as a white solid.

LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>)
<NUM>H NMR (<NUM>, DMSO-d<NUM>) δ ppm <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>) <NUM> - <NUM> (m, <NUM>)
The Intermediate in the following table was prepared from reagents reported below by using methods analogous to Intermediate R1.

To a solution of (R)-<NUM>-(<NUM>-chlorophenyl)ethyl (<NUM>-(<NUM>-amino-<NUM>-methylpyridin-<NUM>-yl)-<NUM>-methylisoxazol-<NUM>-yl)carbamate (Intermediate L1, <NUM>, <NUM> mmol) in MeCN (<NUM>), (-)-trans-<NUM>,<NUM>-cyclohexanedicarboxylic anhydride (<NUM>, <NUM> mmol) was added. The mixture was stirred at RT for <NUM>. The solvent was removed under reduced pressure and the residue was purified by reversed-phase flash chromatography using a gradient of MeCN in acidic H<NUM>O (+ <NUM>% HCOOH) from <NUM>% to <NUM>% affording the title compound (<NUM>, <NUM> mmol, <NUM>% yield).

LC-MS (ESI): m/z (M+<NUM>): <NUM> (Method <NUM>)
<NUM>H NMR (<NUM>, DMSO-d<NUM>) δ ppm <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (d, J=<NUM>, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> (s, <NUM>), <NUM> (s, <NUM>), <NUM> - <NUM> (m, <NUM>), <NUM> - <NUM> (m, <NUM>)
The Examples in the following table were prepared from reagents reported below following similar procedures as for Example <NUM>.

To a solution of Trans-methyl <NUM>-((<NUM>-(<NUM>-((((R)-<NUM>-(<NUM>-chlorophenyl)ethoxy)carbonyl) amino)-<NUM>-methyl-<NUM>-pyrazol-<NUM>-yl)phenyl)carbamoyl)cyclohexane-<NUM>-carboxylate
(Intermediate Q1, <NUM>, <NUM> mmol) in THF (<NUM>)/ Water (<NUM>), LiOH (<NUM>, <NUM> mmol) was added and the mixture was stirred at r. The solvent was removed under reduced pressure and the residue was dissolved in water, then HCl 6N was added up to acidic pH. The precipitate was filtered , dried and submitted to chiral semipreparative SFC.

Conditions: Column: Whelk <NUM> (R,R) (25x2 cm), <NUM>; Modifier: (Methanol + <NUM>% isopropylamine) <NUM>%; Flow rate <NUM>/min; UV detection: <NUM>; Loop: <NUM>µl.

To a solution of Cis-methy-<NUM>-((<NUM>-(<NUM>-((((R)-<NUM>-(<NUM>-chlorophenyl)ethoxy)carbonyl) amino)thiophen-<NUM>-yl)phenyl)carbamoyl)cyclohexane-<NUM>-carboxylate (Intermediate R1, <NUM>, <NUM> mmol) in MeOH (<NUM>)/ Water (<NUM>), LiOH (<NUM>, <NUM> mmol) was added and the mixture was stirred at r. HCl 1N was added up to acidic pH and the mixture was extracted with EtOAc. The organic phase was dried over Na<NUM>SO<NUM> and concentrated under reduced pressure The residue was purified by flash chromatography using a gradient of MeOH in DCM from <NUM>% to <NUM>% affording the title compound (<NUM>, <NUM> mmol, <NUM>% yield).

Example <NUM> was submitted to semipreparative SFC.

Conditions: Column: Chiralpak AD-H (<NUM> x <NUM>), <NUM>; Modifier: Methanol <NUM>%; Flow rate <NUM>/min; UV detection: <NUM>; Loop: <NUM>µl.

To a solution of Trans-methy-<NUM>-((<NUM>-(<NUM>-((((R)-<NUM>-(<NUM>-chlorophenyl)ethoxy)carbonyl) amino)thiophen-<NUM>-yl)phenyl)carbamoyl)cyclohexane-<NUM>-carboxylate (Intermediate Q2, <NUM>, <NUM> mmol) in THF (<NUM>)/ MeOH (<NUM>)/ Water (<NUM>), LiOH (<NUM>, <NUM> mmol) was added and the mixture was stirred at r. HCl 1N was added up to acidic pH and the mixture was extracted with EtOAc. The organic phase was dried over Na<NUM>SO<NUM> and concentrated under reduced pressure. The residue was purified by flash chromatography using a gradient of MeOH in DCM from <NUM>% to <NUM>% affording the title compound (<NUM>, <NUM> mmol, <NUM>% yield).

To a solution of Trans-methyl <NUM>-((<NUM>-(<NUM>-((((R)-<NUM>-(<NUM>-chlorophenyl)ethoxy)carbonyl)amino)-<NUM>-methylisoxazol-<NUM>-yl)-<NUM>-methylpyridin-<NUM>-yl)carbamoyl)cyclohexane-<NUM>-carboxylate (Intermediate Q3, <NUM>, <NUM> mmol) in THF (<NUM>) and Water (<NUM>), LiOH (<NUM>, <NUM> mmol) was added and the mixture was stirred at <NUM> for <NUM>. HCl 1N was added till acidic pH and the mixture was extracted with EtOAc. The organic layer was dried over Na<NUM>SO<NUM> and concentrated under reduced pressure. The residue was purified by flash chromatography using a gradient of MeOH in DCM from <NUM>% to <NUM>% and then submitted to semipreparative SFC.

Conditions: Column: Chiralpak AD-H (<NUM> x <NUM>), <NUM>; Modifier: (Ethanol + <NUM>% trifluoroacetic acid) <NUM> %; Flow rate <NUM>/min; UV detection: <NUM>; Loop: <NUM>µl. The absolute stereochemistry (1R,2R) was assigned by comparing chiral HPLC with Example <NUM>.

To a solution of Cis-methyl <NUM>-((<NUM>-(<NUM>-((((R)-<NUM>-(<NUM>-chlorophenyl)ethoxy)carbonyl)amino)-<NUM>-methylisoxazol-<NUM>-yl)-<NUM>-methylpyridin-<NUM>-yl)carbamoyl)cyclohexane-<NUM>-carboxylate (Intermediate R2, <NUM>, <NUM> mmol) in THF (<NUM>)/ Water (<NUM>), LiOH (<NUM>, <NUM> mmol) was added and the mixture was stirred at r. Volatiles were removed under reduced pressure,the residue was dissolved in water and HCl 3N was added till acidic pH. The solvent was removed under reduced pressure and the residue was purified by reversed-phase flash chromatography using a gradient of MeCN in acid H<NUM>O (+ <NUM>% HCOOH) from <NUM>% to <NUM>% and then submitted to semipreparative SFC.

Conditions: Column: Chiralpak IC (25x0. <NUM>), <NUM>; Modifier: Methanol <NUM>%; Flow rate <NUM>/min; UV detection: <NUM>; Loop: <NUM>µl.

Compound (<NUM>,<NUM>)-<NUM>-((<NUM>-(<NUM>-(((<NUM>-(<NUM>-fluoropyridin-<NUM>-yl)ethoxy)carbonyl)amino)-<NUM>-methylisoxazol-<NUM>-yl)pyridin-<NUM>-yl)carbamoyl)cyclohexane-<NUM>-carboxylic acid (Example <NUM>, <NUM>, <NUM> mmol) was submitted to chiral chromatography.

Conditions: Column: Chiralpak AD-H (<NUM> x <NUM>), <NUM>; Modifier: n-Hexane/(Ethanol + <NUM>% trifluoroacetic acid) <NUM>/<NUM> % v/v; Flow rate <NUM>/min; UV detection: <NUM>; Loop: <NUM>µl.

Compound (<NUM>,<NUM>)-<NUM>-((<NUM>-(<NUM>-(((<NUM>-(<NUM>-fluoropyridin-<NUM>-yl)ethoxy)carbonyl)amino)-<NUM>-methylisoxazol-<NUM>-yl)-<NUM>-methylpyridin-<NUM>-yl)carbamoyl)cyclohexane-<NUM>-carboxylic acid (Example <NUM>, <NUM>, <NUM> mmol) was submitted to chiral semipreparative chromatography. Conditions: Column: Whelk O1 (R,R) (<NUM> x <NUM>), <NUM>; Modifier: n-Hexane/(Ethanol + <NUM>% trifluoroacetic acid) <NUM>/<NUM> % v/v; Flow rate <NUM>/min; UV detection: <NUM>; Loop: <NUM>µl.

To a solution of tert-butyl <NUM>-((<NUM>-(<NUM>-((((R)-<NUM>-(<NUM>-fluorophenyl)ethoxy)carbonyl)amino)-<NUM>-methylisoxazol-<NUM>-yl)-<NUM>-methylpyridin-<NUM>-yl)carbamoyl)cyclohexane-<NUM>-carboxylate (Intermediate J13, <NUM>, <NUM> mmol) in DCM (<NUM>) cooled to <NUM>, a <NUM> solution of HCl in <NUM>,<NUM>-Dioxane (<NUM>, <NUM> mmol) was added dropwise. After stirring overnight at room temperature the solvent was evaporated and the product was purified via pTLC (<NUM>% MeOH in DCM) to give the title compound as a white solid (<NUM>, <NUM> mmol, <NUM>% yield).

The Examples in the following table were prepared from reagents reported below following similar procedures as for Example <NUM>.

To a solution of (R)-<NUM>-(pyridin-<NUM>-yl)ethyl (<NUM>-(<NUM>-amino-<NUM>-methylpyridin-<NUM>-yl)-<NUM>-methylisoxazol-<NUM>-yl)carbamate (Intermediate L2, <NUM>, <NUM> mmol) in DMF (<NUM>), cis-<NUM>,<NUM>-cyclohexanedicarboxylic anhydride (<NUM>, <NUM> mmol) was added and the reaction mixture was stirred overnight at r. Water was added and the product was extracted with DCM (<NUM> x <NUM> ). Combined organic layers were washed with brine (<NUM>), dried over sodium sulphate and evaporated to give <NUM> of crude. The crude was purified via pTLC (<NUM>% MeOH in DCM) to provide the title compound (<NUM>, <NUM> mmol, <NUM>% yield).

To a solution of methyl (<NUM>,<NUM>)-<NUM>-((<NUM>-(<NUM>-((((R)-<NUM>-(<NUM>-chlorophenyl)ethoxy)carbonyl)amino)-<NUM>-methyl-<NUM>H-<NUM>,<NUM>,<NUM>-triazol-<NUM>-yl)-<NUM>-fluoropyridin-<NUM>-yl)carbamoyl)cyclohexane-<NUM>-carboxylate (Intermediate J38, <NUM>, <NUM> mmol) in THF (<NUM>) and Water (<NUM>), a solution of LiOH 1N (<NUM>, <NUM> mmol) was added and the mixture was stirred at r. HCl 1N was added till acidic pH and the mixture was extracted with EtOAc. The organic layer was dried over Na<NUM>SO<NUM> and concentrated under reduced pressure. The residue was purified via reverse phase flash chromatography using a gradient of MeCN (<NUM>% HCOOH) in acidic water (<NUM>% HCCOH) from <NUM> to <NUM>% to afford the title compound (<NUM>, <NUM> mmol, <NUM>% yield).

The effectiveness of compounds of the present invention as LPA1 antagonists can be determined at the human recombinant LPA1 expressed in CHO cells, using a FLIPR assay in <NUM> well format.

CHO-hLPA1 cell lines are cultured in a humidified incubator at <NUM>% CO2 in DMEM/F-<NUM> (<NUM>:<NUM>) MIXTURE with <NUM> Glutamax, supplemented with <NUM>% of Foetal Bovine Serum, <NUM> Sodium Pyruvate, <NUM> Hepes and 1X Penicillin/Streptomycin. CHO hLPA1 cells are seeded into black walled clear-bottom <NUM>-well plates (#<NUM>, Greiner Bio-One GmbH) at a density of <NUM>,<NUM> cells per well in <NUM>µl culture media and grown overnight in a <NUM> humidified CO2-incubator. Serial dilutions (<NUM>:<NUM> or <NUM>:<NUM>, <NUM> points CRC) of compounds are performed in <NUM>% DMSO at 200X the final concentration. The compounds are diluted <NUM>:<NUM> prior to the experiment with Assay Buffer (<NUM> HEPES, <NUM> NaCl, <NUM> KCl, <NUM> glucose, <NUM> MgCl2 and <NUM> CaCl2, pH <NUM> containing <NUM>% Pluronic F-<NUM>) to obtain a solution corresponding to <NUM>-fold the final concentration in the assay (4X, <NUM>% DMSO). The final concentration of DMSO in the assay will be <NUM>% in each well. Medium is removed by aspiration and cells are then incubated with <NUM>µl of a loading solution containing <NUM> of the cytoplasmic Ca2+ indicator Cal-<NUM> AM in Assay Buffer containing <NUM> probenecid for <NUM> at <NUM> 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, <NUM>µl/well of 4X antagonists' solution was added onto the cells. After <NUM> pre-incubation (at <NUM>), <NUM>µl/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 IC<NUM> 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 "<NUM>% inhibition", while the raw data obtained in negative controls, i.e. in the absence of compounds and stimulating with LPA EC80, are set as "<NUM>% inhibition".

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

The results for individual compounds are provided below in Table <NUM> and are expressed as range of activity.

As it can be appreciated, all the compounds of Table <NUM> show an antagonist activity on LPA1 receptor. In fact, it can be recognized that the symbol + indicate a good and sufficient level of activity, which can be even increased up to +++, thus confirming the high activity receptor LPA1 of the compounds of the invention.

BSEP inhibition was evaluated using cryopreserved human hepatocytes (Plateable Cryopreserved Human Hepatocytes, BIOIVT) cultured for <NUM> day between two layer of collagen (sandwich configuration). In this culture condition, hepatocytes express relevant transporters including BSEP and retain the bile canalicular structure.

On day <NUM> of culture, hepatic cells are ready for the assay and the biliary clearance of Taurocolic Acid (TCA), a known BSEP substrate, can be exstimated in presence and in absence of compound of interest.

The sample solution was prepared dissolving test compound and TCA in DMSO and then diluted in the Assay Buffer: Hank's Balance Salt Solution (HBSS+) warmed at <NUM> before use, to give the <NUM> TCA working solution with and without <NUM> of test compound. Hepatocytes were incubated for <NUM> minutes with these working solutions allowing TCA to be excreted into bile. At the end of incubation, the working solution was aspirated, and the content of the bile was collected through the addition of HBSS Modified without Ca2+/Mg2+ (HBSS-). The presence of Ca2+ in the buffer is required to maintain the integrity of the tight junctions, the diffusional barrier between the canalicular lumen and extracellular space. Instead, incubation of cells in Ca2+ -free buffer disrupts the tight junctions and opens the bile canalicular structures, allowing the bile content to be released and collect for HPLC-MS/MS analysis.

The in vitro biliary clearance of TCA incubated with and without test compounds is calculated according to the following formula: <MAT> where <MAT> <MAT>.

The inhibition of BSEP was calculated as percentage of inhibition of TCA biliary clearance in presence of compound of interest, according to the following formula: <MAT>.

The results for individual compounds are provided below in Table <NUM>.

The permeability of the compounds of the present invention was evaluated performing the assy on Caco-<NUM> cells monolayers (human colon adenocarcinoma immortalized cell) by measuring the transport of compound (absorption and secretion) in both directions: apical to basolateral direction (A>B) and basolateral to apical (B>A) with and without PgP inhibitor (Elacridar).

The cells, purched from ReadyCell in <NUM> well format (Cod. KRECE-CCR50), were cultured by the supplier for <NUM> day on transwell supports in DMEM <NUM>/L glucose culture medium supplemented with Fetal Bovin Sierum (<NUM>%), Glutamine <NUM> (<NUM>%) and Penicillin <NUM> U/ml- <NUM>/ml Streptomycin (<NUM>%).

On day <NUM> of colture, cell monolayers integrity was verified by measuring the trans-epithelial electric resistance (TEER) using the EVOM equipment (Endohm, WPI, Germany) and studying the apparent permeability (Papp) of reference compounds (Sulpiride and Metoprolol). Furthermore, as a control, the Talinolol <NUM> (Pgp efflux substrate) with and without Elacridar in both directions was used.

The sample solution was prepared dissolving test compound in DMSO at the concentration of <NUM> and then diluted in the Assay Buffer (Hank's Balance Salt Solution) warmed at <NUM> before use, to give the <NUM> Compound working solution with and without <NUM> Elacridar. These working solutions were added to donor compartment (apical for A>B direction and basolateral for B>A direction ) and Assay Buffer (Hank's Balance Salt Solution) to the receiver compartment ( basolateral for A>B direction and apical for B>A direction ). The plate was incubated at <NUM> for <NUM>, all incubation were conducted in triplicates. At the end of incubation, samples from donor and receiver compartments were collected for HPLC-MS/MS analyses.

The permeability coefficients (Papp) in both directions: apical to basolateral (A>B) and basolateral to apical (B>A) with and without PgP inhibitor (Elacridar) was calculated in nm/sec, using the following equation: <MAT> where:.

The activity of comparative Example A as has been tested in the in vitro assay for the determination of activity on LPA1 receptor as described above along with the BSEP and permeability assays.

Differently from the compounds of formula (I) of the present invention, the comparative Example A shows a passive permeability < <NUM> and thus not suitable for an oral administration and a BSEP inhibition at <NUM> greater than <NUM>%, said inhibition cannot be considered acceptable for a drug candidate.

The activity of comparative Example B as has been tested in the in vitro assay for the determination of activity on LPA1 receptor as described above.

Differently from the compounds of formula (I) of the present invention, the comparative Example B shows an IC<NUM> greater than <NUM> and thus the compound is inactive on receptor LPA1.

Claim 1:
A compound of formula (I)
<CHM>
wherein X is CR<NUM>, -CH- or N,
A is selected from the group consisting of
<CHM>
R<NUM> is selected from the group consisting of aryl, (C<NUM>-C<NUM>)cycloalkyl, heterocycloalkyl, heteroaryl and (C<NUM>-C<NUM>)alkyl wherein any of such aryl, heteroaryl, cycloalkyl, heterocycloalkyl and alkyl may be optionally substituted by one or more groups selected from (C<NUM>-C<NUM>)alkyl, halo, (C<NUM>-C<NUM>)haloalkyl, CN, -O(C<NUM>-C<NUM>)alkyl, - NR<NUM>R<NUM>;
R<NUM> is H or (C<NUM>-C<NUM>)alkyl;
R<NUM> is H or (C<NUM>-C<NUM>)alkyl,
R<NUM> is H or (C<NUM>-C<NUM>)alkyl.
R<NUM> is H or selected from the group consisting of (C<NUM>-C<NUM>)alkyl, halo and CN;
R<NUM> and R<NUM> are at each occurrence independently H or selected from the group consisting of (C<NUM>-C<NUM>)alkyl, (C<NUM>-C<NUM>)haloalkyl and halo, or
R<NUM> and R<NUM> may form together with the nitrogen atom to which they are attached a <NUM>-<NUM> membered saturated heterocyclic ring system optionally containing a further heteroatom selected from N, S and O, said heterocyclic ring system may be optionally substituted by one or more groups selected from (C<NUM>-C<NUM>)alkyl, (C<NUM>-C<NUM>) haloalkyl and halo,
with the proviso that when A is
<CHM>
XisN.