IT201800005598A1 - OXADIAZOLS AS ANTAGONISTS OF THE FXR RECEPTOR - Google Patents

Description

DESCRIPTION

of the patent for industrial invention entitled: "OXADIAZOLS AS ANTAGONISTS OF THE FXR RECEPTOR"

Background of the invention

The present invention relates to oxadiazole derivatives and related uses, in particular in the treatment and / or prevention of FXR-mediated diseases.

The farnesoid X receptor (FXR, also known as BAR, NR1H4) is a ligand-dependent transcription factor and belongs to the super-family of nuclear hormone receptors.

Highly expressed in enterohepatic tissues (liver and intestine), FXR regulates bile acid homeostasis, lipoprotein, glucose metabolism, liver regeneration and cardiovascular disorders (Mangelsdorf, D. J. et al. Cell 1995, 83; 835; Forman, B. M. et al. Cell 1995, 81, 687; Makishima, M. et al. Science 1999, 284, 1362; Parchi, D. J. et al. Science 1999, 284, 1365; Wang, H. et al. Mol Cell 1999, 3, 543; Kuipers, F. et al. Rev. Endocr. Metab.

Disord. 2004, 5.319; Zhang et al. Proc. Natl. Acad. Sci. USA 2006, 103, 1006; Matsubara, T. et al. Mol. Cell. Endocrinol.

2013, 368, 17; Claudel, T. et al. Arterioscler. Thromb. Vasc. Biol. 2005, 25, 2020). Specific bile acids bind and activate FXR, the most potent being chenodeoxycholic acid (CDCA), which is an endogenous ligand of FXR and the primary bile acid found in human bile.

Various potent and selective FXR agonists, belonging to both steroidal and non-steroidal chemical classes, such as 6-ECDCA, GW4064 and Fexaramine, have been reported in recent years. Indeed, as the research field of FXR agonists has expanded, various side effects have arisen from advanced clinical trials with serum lipid dysregulation observed in patients with diabetes and hepatic steatosis (Fiorucci S. et al. Expert Opin Ther Targets 2014, 18, 1449-59). In addition, FXR agonists interfere with the ability of the constitutive androstane receptor to regulate the MRP-4 transporter (Renga B. et al. Biochim Biophys Acta 2011, 3, 157-65) in hepatocytes and this effect worsens the injury. liver in obstructive cholestasis.

Recent studies have reported that antagonizing the activity of FXR may have the potential effect of lowering the levels of total cholesterol, triglycerides and glucose, focusing on the potential application of FXR antagonists in the treatment of metabolic dysregulation. In addition, recent findings have supported the notion that FXR gene ablation protects against hepatocyte injury caused by bile duct ligation (BDL) by affirming the usefulness of FXR antagonists in the treatment of some forms of cholestasis.

Few FXR antagonists have been described so far with very limited structural diversity. The main contribution resulted from the target-oriented decoding of natural marine and terrestrial compounds, with the identification of several steroid scaffolds with promising pharmacological properties. Among these, naturally occurring glyco-β-muricolic acid (GβMCA) has proven itself as a new strategy for the treatment of obesity, NAFLD and insulin resistance (WO2015017813). In the setting of non-steroidal chemotypes, only isoxazole and pyrazole / pyrazolone were identified as privileged scaffolds in the FXR antagonism (WO2015116856). Therefore, structurally novel non-steroidal FXR antagonists represent an important direction in the field.

The aim of the present invention is the identification of new compounds which act as FXR antagonists.

Said object is achieved by the present invention, relative to the selective FXR antagonists containing the 1,2,4-oxadiazole chemical scaffold according to claim 1, to a pharmaceutical composition according to claim 8, to the uses of claims 9 and 10. Forms of preferred embodiments are disclosed within the dependent claims.

The following paragraphs provide the definitions of the various chemical fractions of the compounds according to the invention and are intended to be applied uniformly in the entire specification and in the claims unless a definition otherwise expressly set out does not provide a broader definition.

The term "alkyl", as used herein, refers to saturated aliphatic hydrocarbon groups. This term includes linear (unbranched) chains or branched chains.

Non-limiting examples of alkyl groups according to the invention are, for example, methyl, ethyl, propyl, isopropyl, n-butyl, iso-butyl, tert-butyl, n-pentyl, iso-pentyl, n-hexyl and the like.

The term "pharmaceutically acceptable salts" refers to salts of the compounds of Formula (I) identified below that maintain the desired biological activity and have been accepted by the regulatory authorities.

As used herein the term "salt" refers to any salt of a compound according to the present invention prepared from an inorganic or organic acid or base and internally formed salts. Typically, such salts exhibit a physiologically acceptable anion or cation.

Furthermore, the compounds of Formula (I) can form an acid addition salt or a salt with a base, depending on the type of substituents, and such salts are included in the present invention as long as they are pharmaceutically acceptable salts.

Examples of such salts include, but are not limited to acid addition salts formed with inorganic acids, salts formed with organic acids.

The compounds of formula (I) containing acidic protons can be converted into their non-toxic, therapeutically active basic addition salt forms, for example metal or amine salts, by treatment with appropriate organic and inorganic bases.

Physiologically or pharmaceutically acceptable salts are particularly suitable for medical applications due to their greater solubility in water than the parent compound.

Pharmaceutically acceptable salts can also be prepared from other salts including other pharmaceutically acceptable salts of the compounds of Formula (I) using conventional methods.

Those skilled in the art of organic chemistry will appreciate that many organic compounds can form complexes with solvents in which they are reacted or from which they are precipitated or crystallized. These complexes are known as "solvates". For example, a complex with water is known as a "hydrate". The solvates of the compounds of the invention fall within the scope of the invention. The compounds of Formula (I) can be readily isolated in association with the solvent molecules by crystallization or evaporation of an appropriate solvent to give the corresponding solvates.

The compounds of Formula (I) can be in crystalline form. In some embodiments, the crystalline forms of the compounds of Formula (I) are polymorphic.

The present invention also includes isotopically labeled compounds, which are identical to those set forth in Formula (I) et seq, but differ in that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or by the mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the invention and the related pharmaceutically acceptable salts include isotopes of hydrogen, carbon, nitrogen, oxygen, such as <2> H, <3> H, <11> C, <13> C, < 14> C, <15> N, <17> O, <18> O.

The compounds of the present invention and the pharmaceutically acceptable salts of said compounds which contain the aforementioned isotopes and / or other isotopes of other atoms are within the scope of the present invention. Isotopically labeled compounds of the present invention, for example those in which radioactive isotopes such as <3> H, <14> C are incorporated, are useful in tissue destruction assays of drugs and / or substrates. Tritiated isotopes, i.e. <3> H, and carbon-14, i.e. <14> C, are particularly preferred for their ease of preparation and detectability. The <11> C isotope is particularly useful in PET ("Positron Emission Tomography"). Furthermore, substitution with heavier isotopes such as deuterium, i.e. <2> H, can produce certain therapeutic advantages resulting from greater metabolic stability, e.g. increased half-life in vivo or reduced dosage requirements and thus may be preferred in some circumstances. Isotopically labeled compounds of Formula (I) of this invention can generally be prepared by carrying out the procedures illustrated in the Schemes and / or Examples below, by replacing a nonisotopically labeled reagent with a readily available isotopically labeled reagent.

Certain groups / substituents included in the present invention can be present as isomers. Consequently, in some embodiments, the compounds of Formula (I) may have asymmetric carbon atoms or axial asymmetries in some cases and correspondingly they may exist in the form of optical isomers such as an (R) form, a form (S), and the like. The present invention includes within the scope all such isomers, including racemates, enantiomers and related mixtures.

In particular, all stereoisomeric forms are included within the scope of the present invention, including enantiomers, diastereomers and related mixtures, including racemates and the general reference for compounds of Formula (I) includes all stereoisomeric forms, unless is not otherwise indicated.

In general, the compounds or salts of the invention must be interpreted in such a way as to exclude those compounds (if any) which are chemically unstable, either by themselves or in water, which are clearly not suitable for pharmaceutical use by all routes. oral, parenteral, or otherwise. Such compounds are known to the skilled chemist.

According to a first aspect of the invention, the compounds of Formula (I) are provided:

or pharmaceutically acceptable salts or solvates.

In the compounds of Formula (I):

A1 and A2 are selected from the group consisting of CH2 and NR1 provided that A1 and A2 are not at the same time CH2 or NR1,

R1 is selected by CH3 and (CH2) 2-4-R2,

R2 is selected from the group consisting of H, CH3, COOR3 and CH2OH,

R3 is selected from the group consisting of H and C1-4 alkyl,

R4, R5, R6, R7, R8, R9 and R10 are independently selected from the group consisting of H, C1-4 alkyl, O-C1-4 alkyl, halogen and OH.

n is selected between 1 and 2.

According to a first embodiment, n is 1. Such compounds can have the formula (Ia):

wherein R1, R2 R3, R4, R5, R6, R7, R8, R9 and R10 are as defined above.

According to a second embodiment, n is 2. These compounds can have the formula (Ib)

or the formula (Ic)

wherein R1, R2 R3, R4, R5, R6, R7, R8, R9 and R10 are as defined above.

Preferably, R1 is (CH2) 3-R2.

Preferably, R2 is COOR3.

Preferably, R3 is a methyl group.

Preferably each of R4, R5, R6, R7, R8, R9 and R10 is hydrogen.

According to a third embodiment of the invention, the compounds of Formula (I) can be selected from the group consisting of:

The compounds exemplified in this invention can be prepared from readily available starting materials using the following general methods and procedures for example exemplified in Michael B. Smith - March's Advanced Organic Chemistry: reactions, mechanisms, and structure - 7 <a> edition, John Wiley & Sons Inc., 2013.

It is well known to an average person skilled in the art that the transformation of a chemical function into another may require that one or more reactive centers in the compound containing this function be protected in order to avoid unwanted side reactions. The protection of such reactive centers, and the subsequent de-protection at the end of the synthetic transformations, can be achieved by following the standard procedures described for example in Peter G.M. Wuts - Green's Protective Groups in Organic Synthesis, Fifth Edition, John Wiley & Sons Inc., 2014.

It will be appreciated that in situations where typical or preferred experimental conditions are reported (i.e. reaction temperatures, time, moles of reagents, solvents, etc.), other experimental conditions may also be used unless otherwise indicated. Optimal best reaction conditions may vary with the particular reagents or solvents used, but such conditions can be determined by one skilled in the art, using routine optimization procedures.

The synthesis of a compound of Formula (I), according to the synthesis processes described below, can be carried out in a stepwise manner, whereby each intermediate is isolated and purified by standard purification techniques such as, for example, column chromatography, before carrying out the next reaction. Alternatively, two or more steps of the synthesis sequence can be carried out in a so-called "one-pot" procedure, as known in the art, whereby only the compound resulting from the two or more steps is isolated and purified.

The compounds of Formula (I), prepared by the methods described hereinafter, can be treated or purified by conventional techniques or means, for example by filtration, distillation, chromatography, recrystallization and relative combination.

The salts of the compounds of Formula (I) can be prepared by reacting a basic compound with the desired acid in solution, or by reacting an acid compound with the desired base in solution.

A second aspect of the present invention relates to a pharmaceutical composition comprising a compound of Formula (I) in which:

A1 and A2 are selected from the group consisting of CH2 and NR1 provided that A1 and A2 are not both at the same time CH2 or NR1,

R1 is selected from the group consisting of H, CH3 and (CH2) 2-

4-R2,

R2 is selected from the group consisting of H, COOR3 and CH2OH,

R3 is selected from the group consisting of H and C1-4 alkyl,

R4, R5, R6, R7, R8, R9 and R10 are independently selected from the group consisting of H, C1-4 alkyl, O-C1-4 alkyl, halogen and OH.

n is selected between 1 and 2 and at least one pharmaceutically acceptable excipient.

An expert in the art is aware of a whole variety of such excipients compounds suitable for formulating a pharmaceutical composition.

The compounds of the invention, together with a conventionally employed excipient, can be placed in the form of pharmaceutical compositions and relative unit dosages, and in such form they can be used as solids, such as tablets or filled capsules, or liquids such as solutions, suspensions, emulsions, elixirs or capsules filled with it, all for oral use or in the form of sterile injectable solutions for parenteral administration (including subcutaneous and intravenous use).

Such pharmaceutical compositions and related unit dosage forms may comprise ingredients in conventional proportions, with or without additional active compounds or ingredients, and such unit dosage forms may contain any suitable effective amount of the active ingredient proportionate to the daily dosage range to be employed. planned.

Pharmaceutical compositions containing a compound of this invention can be prepared in a manner well known in the pharmaceutical art and comprise at least one active compound. In general, the compounds of this invention are administered in a pharmaceutically effective amount. The amount of the compound actually administered will typically be determined by a physician, in light of the relevant circumstances, including the condition being treated, the route of administration chosen, the actual compound administered, the age, weight and response of the individual patient. the severity of the patient's symptoms and the like.

The pharmaceutical compositions of the present invention can be administered by a variety of routes including oral, rectal, subcutaneous, intravenous, intramuscular, intranasal and pulmonary routes. The compositions for oral administration can take the form of liquid solutions or suspensions in bulk, or powders in bulk. Most commonly, however, the compositions are presented in unit dosage forms to facilitate precise dosing. The term "unit dosage forms" refers to physically discrete units suitable as unit dosages for humans and other mammals, each unit containing a predetermined amount of active material calculated to produce the desired therapeutic effect in association with a suitable pharmaceutical excipient . Typical unit dosage forms include pre-filled vials or syringes, pre-measured with the liquid compositions or pills, tablets, capsules or the like in the case of solid compositions.

Liquid forms suitable for oral administration may include a suitable aqueous or non-aqueous vehicle with buffers, suspending and dispersing agents, dyes, flavoring and the like. Solid forms may include, for example, any of the following ingredients, or compounds of a similar nature: a binder such as microcrystalline cellulose, tragacanth or gelatin; an excipient such as starch or lactose, a disintegrating agent such as alginic acid, Primogel or corn starch; a lubricant such as magnesium stearate; a fluidifier such as colloidal silicon dioxide; a sweetening agent such as sucrose or saccharin or a flavoring agent such as peppermint, methyl salicylate or orange flavoring.

The injectable compositions are typically based on injectable sterile saline or phosphate buffered saline or other injectable carriers known in the art.

The pharmaceutical compositions can be in the form of tablets, pills, capsules, solutions, suspensions, emulsions, powders, suppositories and as extended release formulations.

If desired, the tablets can be coated by standard aqueous or non-aqueous techniques. In some embodiments, such compositions and preparations may contain at least 0.1 percent active compound.

The percentage of active compound in these compositions can obviously be varied and can conveniently be between about 1 percent and about 60% of the weight of the unit. The amount of active compound in such therapeutically useful compositions is such that a therapeutically active dosage will be obtained. The active compounds can also be administered intranasally as, for example, liquid drops or sprays.

The tablets, pills, capsules and the like may also contain a binder such as tragacanth, acacia, corn starch or gelatin, excipients such as calcium phosphate, a disintegrating agent such as corn starch, potato starch, alginic acid, a lubricant such as stearate of magnesium and a sweetening agent such as sucrose, lactose or saccharin. When a unit dosage form is a capsule, it may contain in addition to the materials of the above type, a liquid carrier such as a fatty oil. Various other materials may be present as coatings or to modify the physical form of the metering unit. For example, the tablets can be coated with shellac, sugar or both. A syrup or elixir may contain, in addition to the active ingredient, sucrose as a sweetening agent, methyl and propyl parabens as preservatives, a color and a flavoring agent such as cherry or orange flavoring. To avoid disruption during transit through the upper portion of the gastrointestinal tract, the composition is an enteric-coated formulation.

Compositions for pulmonary administration include, but are not limited to, dry powder compositions composed of the powder of a compound of Formula (I) or a related salt and the powder of a suitable carrier and / or lubricant. The compositions for pulmonary administration can be inhaled by any dry powder inhaler device known to one skilled in the art.

The administration of the compositions is performed within the framework of a protocol and at a dosage sufficient to reduce inflammation and pain in the subject. In some embodiments, in the pharmaceutical compositions of the present invention the active ingredient or ingredients are generally formulated in dosage units. The dosage unit may contain from 0.1 to 1000 mg of a compound of Formula (I) per dosage unit for daily administration.

In some embodiments, the effective quantities for a specific formulation will depend on the severity of the disease, disorder or condition, the previous therapy, the individual's state of health and the response to the drug. In some embodiments, the dose is in the range of 0.001% by weight to about 60% by weight of the formulation.

When used in combination with one or more different active ingredients, the compound of the present invention and the other active ingredient can be used in lower doses than when each is used individually.

Regarding formulations with respect to any variety of routes of administration, methods and formulations for drug administration are illustrated in Remington's Pharmaceutical Sciences, 17th edition, Gennaro et al. Eds, Mack Publishing Co., 1985 and Remington's Pharmaceutical Sciences, Gennaro AR ed. 20 <a> edition, 2000, Williams & Wilkins PA, USA, and Remington: The Science and Practice of Pharmacy, 21 <a> Edition, Lippincott Williams & Wilkins Eds., 2005; and in Loyd V. Allen and Howard C. Ansel, Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, 10 <a> Edition, Lippincott Williams & Wilkins Eds., 2014.

The components described above for orally administered or injectable compositions are purely representative.

The compounds of this invention can also be administered in sustained release forms or from sustained release drug delivery systems.

A third aspect of the present invention relates to compounds of Formula (I), including those compounds of Formula (I) in which R1 is H, or the pharmaceutical composition as illustrated above, for use as a medicament.

Particularly preferred compounds for use as a medicinal product are:

In particular the compounds of Formula (I) as illustrated above, including those in which R1 is H, or the relative pharmaceutical composition, can be used for the prevention and / or treatment of a disorder selected from the group consisting of gastrointestinal disorders, liver diseases, cardiovascular and vascular diseases, pulmonary and metabolic diseases, infectious diseases, cancer, kidney disorders, inflammatory disorders including immune-mediated disorders and neurological disorders.

In one embodiment, the immune-mediated inflammatory disorders include autoimmune disorders such as systemic lupus erythematosus, rheumatoid arthritis, Sjögren's syndrome, scleroderma also known as systemic sclerosis, spondyloarthritis, vasculitis, sarcoidosis, Mediterranean fever and other inherited autoinflammatory diseases, polymyositis and dermatomyositis, Behcet's syndrome.

In one embodiment, infectious diseases are selected from the group of Acquired Immuno-Deficiency Syndrome (AIDS) and related disorders, virus B and virus C infections.

In one embodiment, neurological disorders include Alzheimer's disease and other forms of dementia, Parkinson's disease and other movement disorders, amyotrophic lateral sclerosis and other neuromotor disorders, multiple sclerosis and other demyelinating diseases, ischemic stroke, myasthenia and muscular dystrophy.

In one embodiment, liver disorders include primary biliary cirrhosis (PBC), cerebrotendinous xanthomatosis (CTX), primary sclerosing cholangitis (PSC), drug induced cholestasis , intrahepatic cholestasis of pregnancy, cholestasis associated with parenteral nutrition, cholestasis associated with bacterial hyperproliferation or sepsis, autoimmune hepatitis, chronic viral hepatitis, alcoholic liver disease, nonalcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (" nonalcoholic steatohepatitis ”, NASH), liver transplantation, congenital hepatic fibrosis, granulomatous liver disease, intra- or extrahepatic malignancy, Wilson's disease, hemochromatosis, and alpha 1-antitrypsin deficiency.

In one embodiment, gastrointestinal disorders include inflammatory bowel disease (IBD) (including Crohn's disease, ulcerative colitis and indeterminate colitis), irritable bowel syndrome (IBS) , bacterial hyperproliferation, acute and chronic pancreatitis, malabsorption, post-radiation colitis and microscopic colitis.

In one embodiment, renal disorders include diabetic nephropathy, hypertensive nephropathy, chronic glomerulonephritis including chronic transplant glomerulonephritis, chronic tubulointerstitial diseases and vascular disorders of the kidney.

In one embodiment, cardiovascular diseases include atherosclerosis, arteriosclerosis, dyslipidemia, hypercholesterolemia, hypertriglyceridemia, hypertension, also known as arterial hypertension, inflammatory heart disease including myocarditis and endocarditis, ischemic heart disease, stable angina, unstable angina, myobascular disease including ischemic stroke.

In one embodiment, vascular diseases include pulmonary heart disease such as pulmonary hypertension, peripheral arterial disease (PAD), also known as peripheral vascular disease (PVD), peripheral artery occlusive disease, and peripheral arterial disease.

In one embodiment, lung disorders include asthma, cystic fibrosis, obstructive respiratory disease, interstitial lung disease, including, but not limited to, primary or secondary pulmonary fibrosis.

In one embodiment, the metabolic disease is selected from the group of diseases including insulin resistance, metabolic syndrome, type I and type II diabetes, hypoglycemia, disorders of the adrenal cortex including insufficiency of the adrenal cortex. Metabolic diseases also include obesity and conditions associated with bariatric surgery.

In one embodiment, the cancer is selected from the group comprising liver cancer, bile duct cancer types, esophageal cancer, pancreatic cancer, gastric cancer, colorectal cancer, breast cancer, ovarian cancer, and the condition associated with resistance to chemotherapy.

Further characteristics of the present invention will emerge from the following description of some purely illustrative and non-limiting examples.

The following abbreviations are used in the attached examples.

Methylene chloride (CH2Cl2), hydroxylamine hydrochloride (NH2OH), methanol (MeOH), potassium carbonate (K2CO3), sodium sulphate (Na2SO4), N, N-diisopropylethylamine (DIPEA), dimethylformamide (DMF), 2- (1H -benzotriazol-1-yl) -1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), lithium bromide (LiBr), sodium bicarbonate (NaHCO3), trifluoroacetic acid (TFA), tetrahydrofuran (THF), lithium hydroxide (LiOH), hydrochloric acid (HCl), ethyl acetate (EtOAc), Nitrogen (N2) and water (H2O), hour (h), ambient temperature (rt), retention time (tR).

Unless otherwise indicated, <1> H NMR was recorded on Varian Inova 400 MHz, using CDCl3 as the solvent, and <13> C NMR was recorded on Varian Inova 100 MHz, using CDCl3 as the solvent.

Examples

EXAMPLE 1 PREPARATION OF OXA7, OXA20, OXA21

The key phase of the synthesis protocol was the preparation of the amidoxime intermediate 2 which in turn is coupled to the corresponding protected N-BOC amino acid. Acid deprotection produced OXA7, OXA20, OXA21 (Scheme 1).

Scheme 1

a) NH2OH HCI, K2CO3, CH3OH, reflux, quantitative yield; b) N-Boc-Inp-OH, DIEA, HBTU, DMF, 80 ° C, 80%; c) TFA, CH2Cl2, 2 hours; d) N-Boc-L-Pip-OH, DIEA, HBTU, DMF, 80 ° C, 78%; e) N-Boc-L-Pro-OH, DIEA, HBTU, DMF, 80 ° C, 83%.

Phase a. Preparation of amidoxime 2

To a solution of 2-naphthonitrile 1 (1 eq.) In anhydrous MeOH, K2CO3 (1.5 mol eq.) And hydroxylamine hydrochloride (2.5 mol eq.) Were added and the mixture was stirred at reflux for 2 hours under nitrogen atmosphere. The resulting solution was then concentrated in vacuo, diluted with water and extracted with CH2Cl2. The organic phases were dried (Na2SO4), filtered and concentrated in vacuo to give amidoxime 2 in quantitative yield, which was subjected to the next phase without any purification.

Example 1A: Preparation of 3- (naphthalen-2-yl) -5- (piperidin4-yl) -1,2,4-oxadiazole (OXA7)

Steps b, c. DIPEA (1.8 mol eq.) Was added to a solution of 2 (1 mol eq.) And N-Boc-isonipecotic acid (1.2 mol eq.) Dissolved in anhydrous DMF. HBTU (1.5 mol eq.), As a coupling agent, was then added to the mixture at room temperature. The mixture was vigorously stirred at 80 ° C for 12 hours, then partitioned between water and EtOAc. The organic layer was collected and washed twice with a saturated LiBr solution, then with a saturated NaHCO3 solution and brine, dried over Na2SO4, filtered and concentrated under reduced pressure. The resulting residue was purified on a silica column using 100% CH2Cl2 to give an intermediate which was deprotected with CH2Cl2: TFA 1: 1 (1 mL) for 2 hours. The resulting residue was purified on a silica column using hexane and EtOAc 99: 1 to give OXA7 in 80% yield.

An analytical sample was further purified by HPLC on Luna Synergi Fusion-RP (4 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (55:45) as eluent (flow rate 1 mL / min, tr = 8.6 min);

OXA7: C17H17N3O

<1> H NMR (500 MHz, CD3OD): δH 8.62 (1H, s), 8.11 (1H, d, J = 8.6 Hz), 8.00 (1H, d, J = 8, 6 Hz), 7.99 (1H, ovl), 7.94 (1H, d, J = 7.2 Hz), 7.60 (1H, ovl), 3.54 (3H, m), 3.26 (2H, m), 2.44 (2H, dd, J = 14.5, 2.8 Hz), 2.19 (2H, m);

δC 181.7, 169.4, 136.1, 134.0, 129.9, 129.8, 128.9 (2C), 128.8 (2C), 128.1, 125.3, 124.6 , 44.0 (2C), 32.9, 27.1 (2C).

Example 1B Preparation of (S) -3- (naphthalen-2-yl) -5- (piperidin-2-yl) -1,2,4-oxadiazole (OXA21)

Steps d, c. OXA21 was prepared starting from 2, under the same operating conditions described in steps b) and c), example 1A using N-Boc-L-pipecolic acid in step d. HPLC purification on Luna Synergi polar-RP (4 μm; 4.6 mm internal diameter x 250 mm) with MeOH / H2O (50:50) as eluent (flow rate 1 mL / min, tR = 14.0 min) produced OXA21.

OXA21 C17H17N3O

δH 8.67 (1H, s), 8.14 (1H, d, J = 8.5 Hz), 8.02 (1H, d, J = 8.7 Hz), 7.98 (1H, ovl) , 7.95 (1H, d, J = 7.4 Hz), 7.62 (1H, ovl), 7.60 (1H, ovl), 4.90 (1H, ovl), 3.60 (1H, ovl), d, J = 12.3), 3.25 (1H, m), 2.51 (1H, m), 2.03 (3H, m), 1.81 (2H, m);

δC 175.9, 169.2, 135.9, 134.1, 129.8, 129.6, 129.2, 128.8 (2C), 127.9, 124.3, 124.2, 52, 9, 45.7, 28.1, 22.5, 22.0. Example 1C. (S) -3- (naphthalen-2-yl) -5- (pyrrolidin-2-yl) -1,2,4-oxadiazole (OXA20)

Steps e, c. OXA20 was prepared starting from 2, under the same operating conditions described in steps b) and c), example 1A using N-Boc-L-proline in step e. HPLC purification on Luna Synergi Fusion-RP (4 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (50:50) as eluent (flow rate 1 mL / min, tR = 11.0 min) produced OXA20.

OXA20 C16H15N3O

δH 8.65 (1H, s), 8.12 (1H, d, J = 8.5 Hz), 8.02 (1H, d, J = 8.6 Hz), 7.99 (1H, ovl) , 7.95 (1H, d, J = 7.1Hz), 7.61 (1H, ovl), 7.60 (1H, ovl), 5.19 (1H, t, J = 7.6Hz) , 3.59 (1H, m), 3.58 (1H, m), 2.69 (2H, m), 2.29 (2H, m);

<13> C NMR (125 MHz, CD3OD) δC 176.3, 168.8, 136.3, 134.3, 130.1, 129.8, 129.2, 129.0 (2C), 128.1 , 124.4, 124.3, 55.6, 47.3, and 24.6.

EXAMPLE 2 GENERAL PROCEDURE FOR N-ALKYLATION ON OXA7

a) acyl bromide, DIPEA in CH3CN, 60 ° C; b) LiOH in THF / H, 0 ° C and therefore the ambient temperature; c) DIBAL-H in THF, 0 ° C; d) alkyl bromide, DIPEA in CH3CN, 60 ° C.

Phase a. OXA7 (1 mol eq.), N, N-diisopropylethylamine (1.5 mol eq.) And methyl ester of bromide Br (CH2) nCOOMe (1.1 mol eq.) In anhydrous acetonitrile were placed in a bottom flask round and stirred at room temperature under nitrogen. After completion of the reaction (monitored by TLC), the resulting solution was concentrated in vacuo, diluted with water and extracted with CH2Cl2. The organic fraction was dried over NaSO4 and the solvent was removed under reduced pressure to give the corresponding N-alkylated esters as crude products.

Step b. Each N-alkylated ester was dissolved in THF / H2O (3: 1) and treated with LiOH hydrate (2 mol eq.) At 0 ° C. The resulting mixture was stirred at rt for 24 hours, followed by treatment with 0.5 N HCl, until pH 7-8 was reached, then partitioned three times with EtOAc. The combined organic extracts were dried over Na2SO4, filtered and concentrated in vacuo to produce the corresponding N-alkylated carboxylic acids.

Phase c. Each N-alkylated ester was dissolved in THF and cooled to -78 ° C under an N2 atmosphere while a solution of diisobutylammonium hydride (1.7 M in toluene, 2 mol eq.) Was added dropwise. The reaction mixture was allowed to slowly warm up to rt and stirred for 48 hours. The reaction was quenched by slow addition of MeOH and then a saturated sodium-potassium tartrate solution was added and stirred for 1 hour. The mixture was partitioned three times and the combined organic extracts dried over Na2SO4.

Phase d. OXA7 (1 mol eq.), N, N-diisopropylethylamine (3 mol eq.), 1-bromopentane (1.5 mol eq.) In anhydrous acetonitrile, were placed in a round bottom flask and stirred at 60 ° C For a night. After completion of the reaction (monitored by TLC), the resulting solution was then concentrated in vacuo, diluted with water and extracted with CH2Cl2. The organic fraction was dried over NaSO4 and the solvent was removed under reduced pressure to give the crude product.

Example 2A. Methyl 4- (4- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) piperidin-1-yl) butanoate (OXA15)

OXA15 was purified by HPLC on Luna Synergi Fusion-RP (4 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (75:25) as eluent (flow rate 1 mL / min, tR = 21.0 min).

OXA15 C22H25N3O3

<1> H NMR (500 MHz, CD3OD): δH 8.61 (1H, s), 8.10 (1H, d, J = 8.5 Hz), 7.99 (1H, ovl), 7.98 (1H, ovl), 7.93 (1H, d, J = 7.6 Hz), 7.59 (1H, ovl), 7.58 (1H, ovl), 3.68 (3H, s), 3 , 14 (1H, m), 3.03 (2H, d, J = 11.3 Hz), 2.44 (2H, t, J = 7.4 Hz), 2.39 (2H, t, J = 7.2 Hz), 2.25 (2H, m), 2.22 (2H, m), 2.03 (2H, m), 1.86 (2H, m);

δC 184.5, 175.8, 168.7, 136.1, 134.0, 129.9, 129.8, 128.9, 128.8 (2C), 128.1, 125.1, 124, 6, 58.9, 53.7 (2C), 52.1, 35.4, 32.6, 30.1 (2C), 22.7.

Example 2B 4- (4- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) piperidin-1-yl) butanoic (OXA16)

OXA16 was purified by HPLC on Luna Synergi Fusion-RP (4 µm; 4.6 mm ID x 250 mm) with MeOH / H2O (7: 3) as eluent (flow rate 1 mL / min, tR = 21.0 min).

OXA16 C21H23N3O3

<1> H NMR (500 MHz, CD3OD): δH 8.61 (1H, s), 8.10 (1H, d, J = 8.6 Hz), 7.99 (2H, d, J = 8, 6 Hz), 7.93 (1H, d, J = 7.5 Hz), 7.59 (1H, ovl), 7.58 (1H, ovl), 3.45 (3H, m), 2.98 (4H, m), 2.43 (4H, m), 2.25 (2H, m), 1.93 (2H, m);

δC 182.4, 169.7, 155.5, 132.6, 130.2 (2C), 128.3 (2C), 127.3, 82.1, 69.9, 55.7, 54.0 , 41.8, 28.3 (3C).

Example 2C. 4- (4- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) piperidin-1-yl) butan-1-ol (OXA17)

OXA17 was purified by HPLC on Luna Synergi Fusion-RP (4 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (75:25) as eluent (flow rate 1 mL / min, tR = 24.0 min).

OXA17 C21H25N3O2

<1> H NMR (500 MHz, CD3OD): δH 8.61 (1H, s), 8.09 (1H, d, J = 8.4 Hz), 7.99 (2H, d, J = 8, 3 Hz), 7.93 (1H, d, J = 8.0 Hz), 7.58 (1H, ovl), 7.57 (1H, ovl), 3.59 (2H, t, J = 5, 9 Hz), 3.18 (1H, m), 3.07 (2H, d, J = 11.4 Hz), 2.47 (2H, t, J = 7.2 Hz), 2.30 (2H , m), 2.22 (2H, m), 2.02 (2H, m), 1.66 (2H, m), 1.60 (2H, m);

δC 184.5, 168.6, 136.1, 134.0, 129.9, 129.8, 128.9, 128.8 (2C), 128.1, 125.0, 124.6, 62.8, 59.7, 53.7 (2C), 35.4, 32.0, 30.2 (2C), 24.5.

2D example. Methyl 3- (4- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) piperidin-1-yl) propanoate (OXA33)

OXA33 was purified by HPLC on Synergi Fusion-RP (4 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (80:20) as eluent (flow rate 1 mL / min, tR = 13.5 min ).

OXA33 C21H23N3O3

<1> H NMR (500 MHz, CD3OD): δH 8.61 (1H, s), 8.10 (1H, d, J = 8.5 Hz), 7.99 (1H, ovl), 7.98 (1H, ovl), 7.93 (1H, d, J = 7.6 Hz), 7.59 (1H, ovl), 7.57 (1H, ovl), 3.70 (3H, s), 3 , 16 (1H, m), 3.02 (2H, d, J = 11.3 Hz), 2.76 (2H, t, J = 7.4 Hz), 2.59 (2H, t, J = 7.2 Hz), 2.31 (2H, m), 2.20 (2H, m), 2.02 (2H, m);

δC 183.6, 174.4, 169.5, 136.1, 134.5, 129.9, 129.8, 128.9, 128.8 (2C), 128.0, 125.5, 124, 6, 54.7, 53.5 (2C), 52.2, 35.4, 32.6, 30.3 (2C).

Example 2E. 3- (4- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) piperidin-1-yl) propan-1-ol (OXA45)

OXA45 was purified by HPLC on Luna Omega Polar C18 (5 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (75:25) as eluent (flow rate 1 mL / min, tR = 13.3 min ).

OXA45 C20H23N3O2

<1> H NMR (500 MHz, CD3OD): δH 8.61 (1H, s), 8.10 (1H, d, J = 8.5 Hz), 7.99 (1H, ovl), 7.98 (1H, ovl), 7.93 (1H, d, J = 7.6 Hz), 7.58 (2H, ovl), 3.64 (2H, t, J = 6.3) 3.17 (1H , m), 3.07 (2H, d, J = 11.2 Hz), 2.56 (2H, t, J = 7.6 Hz), 2.29 (2H, t, J = 11.2) , 2.20 (2H, d, J = 12.4), 2.03 (2H, m), 1.79 (2H, m);

δC 183.6, 169.4, 136.1, 134.5, 129.9 (2C), 128.9, 128.7 (2C), 127.9, 125.5, 124.6, 61.8, 57.1, 53.7 (2C), 35.4, 30.2 (2C), 30.1.

Example 2F. 3- (naphthalen-2-yl) -5- (1-pentylpiperidin-4-yl) -1,2,4-oxadiazole (OXA36)

OXA36 was purified by HPLC on Luna Synergi Fusion-RP (4 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (88:12) as eluent (flow rate 1 mL / min, tR = 11.7 min).

OXA36 C22H27N3O

δH 8.62 (1H, s), 8.11 (1H, d, J = 8.6 Hz), 8.00 (1H, ovl), 7.99 (1H, ovl), 7.93 (1H, d, J = 7.2 Hz), 7.59 (1H, ovl), 7.58 (1H, ovl), 3.14 (1H, m), 3.03 (2H, m), 2.40 (2H, m), 2.21 (4H, m), 2.03 (2H, m), 1.57 (2H, m), 1.37 (2H, m), 1.33 (2H, m), 0, 93 (3H, t, J = 7.1 Hz);

δC 184.5, 168.7, 136.1, 134.0, 129.9, 129.8, 128.9 (2C), 128.8, 128.1, 124.6, 124.3, 53, 7 (2C), 35.4, 30.1 (2C), 14.3.

EXAMPLE 3 GENERAL PROCEDURE FOR N-ALKYLATION ON OXA21 The alkylation on OXA21 with the desired methyl bromide ester Br (CH2) nCOOMe or the desired alkyl bromide Br (CH2) nCH3 was carried out in the same operating condition described respectively in steps a ) and d) of Example 2. Hydrolysis and reduction in correspondence with the methyl ester functional group were carried out in the same operating condition described in steps b) and c) of Example 2.

a) acyl bromide, DIPEA in CH3CN, 60 ° C; b) LiOH in THF / H, 0 ° C and therefore the ambient temperature; c) DIBAL-H in THF, 0 ° C; d) alkyl bromide, DIPEA in CH3CN, 60 ° C.

Example 3A: (S) -methyl 4- (2- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) piperidin-1-yl) butanoate (OXA28)

OXA28 was purified by HPLC on Luna Synergi Fusion-RP (4 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (75:25) as eluent (flow rate 1 mL / min, tR = 31.2 min).

OXA28 C22H25N3O3

δH 8.62 (1H, s), 8.11 (1H, d, J = 8.6 Hz), 8.00 (1H, ovl), 7.99 (1H, ovl), 7.93 (1H, d, J = 7.2 Hz), 7.59 (1H, ovl), 7.58 (1H, ovl), 4.04 (1H, m), 3.56 (3H, s), 3.09 ( 2H, m), 2.40 (2H, m), 2.34 (2H, t, J = 7.1 Hz), 1.98 (2H, m), 1.81 (2H, m), 1, 74 (2H, m), 1.55 (2H, m);

δC 181.3, 175.5, 169.2, 136.2, 134.4, 129.9, 129.8, 128.9 (2C), 128.7, a 128.0, 125.3, 124.6, 59 , 7, 56.1, 52.0, 51.7, 32.2, 31.6, 26.2, 22.9, 22.7.

Example 3B (S) -4- (2- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) piperidin-1-yl) butanoic (OXA39)

OXA39 was purified by HPLC on Luna Synergi Fusion-RP (4 μm; 4.6 mm internal diameter x 250 mm) with MeOH / H2O (7: 3) as eluent (flow rate 1 ml / min, tR = 14.8 min ).

OXA39 C21H23N3O3

δH 8.65 (1H, s), 8.12 (1H, d, J = 8.6 Hz), 8.01 (1H, ovl), 8.00 (1H, ovl), 7.94 (1H, d, J = 7.6Hz), 7.61 (1H, ovl), 7.59 (1H, ovl), 4.16 (1H, dd, J = 6.8, 4.5Hz), 3, 19 (1H, m), 2.55 (2H, m), 2.51 (1H, m), 2.32 (2H, m), 2.03 (2H, m), 1.82 (2H, m ), 1.77 (2H, m), 1.58 (2H, m);

δC 181.2, 178.2, 169.4, 136.4, 134.6, 129.9 (2C), 129.0, 128.9, 128.8, 128.0, 125.0, 124, 7, 59.7, 56.8, 51.7, 33.7, 31.4, 26.1, 23.0, 22.9.

Example 3C. (S) -4- (2- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5yl) piperidin-1-yl) butan-1-ol (OXA41)

OXA41 was purified by HPLC on Luna Synergi Fusion-RP (4 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (75:25) as eluent (flow rate 1 mL / min, tR = 18.7 min).

OXA41 C21H25N3O2

δH 8.63 (1H, s), 8.12 (1H, d, J = 8.6 Hz), 8.01 (1H, d, J = 8.7 Hz), 7.99 (1H, ovl) , 7.94 (1H, d, J = 7.4 Hz), 7.60 (1H, ovl), 7.59 (1H, ovl), 4.08 (1H, br t, J = 5.9 Hz ), 3.52 (2H, t, J = 6.0 Hz), 3.14 (1H, m), 2.46 (2H, m), 2.43 (1H, m), 2.01 (2H , m), 1.89 (1H, m), 1.76 (2H, m), 1.61 (2H, m), 1.54 (1H, m), 1.50 (2H, m);

δC 181.7, 169.4, 136.2, 134.2, 129.9, 129.8, 129.0, 128.8 (2C), 128.1, 125.1, 124.6, 62, 7, 59.6, 57.0, 51.6, 31.4, 31.3, 26.1, 23.8, 22.9.

3D example. (S) -methyl3- (2- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) piperidine-1-yl) propanoate (OXA43)

OXA43 was purified by HPLC on Synergi Hydro-RP (4 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (80:20) as eluent (flow rate 1 mL / min, tR = 17.5 min ).

OXA43 C21H23N3O3

δH 8.63 (1H, s), 8.12 (1H, d, J = 8.6 Hz), 8.01 (1H, d, J = 8.7 Hz), 7.99 (1H, ovl) , 7.94 (1H, d, J = 7.4 Hz), 7.60 (1H, ovl), 7.59 (1H, ovl), 4.13 (1H, br t, J = 5.6 Hz ), 3.63 (3H, s), 3.08 (1H, m), 2.82 (1H, m), 2.74 (1H, m), 2.53 (2H, ovl), 2.52 (1H, ov1), 2.00 (2H, m), 1.82 (1H, m), 1.74 (2H, m), 1.55 (1H, m);

δC 181.7, 174.4, 169.3, 136.2, 134.5, 129.9, 129.8, 128.9, 128.8 (2C), 128.0, 125.3, 124.6, 59.2, 52.5, 52.2, 51.3, 33.0, 31.4, 26.4, 22.9.

Example 3E. (S) -3- (2- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) piperidin-1-yl) propan-1-ol (OXA46)

OXA46 was purified by HPLC on Luna Omega Polar C18 (5 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (78:22) as eluent (flow rate 1 mL / min, tR = 11.0 min ).

OXA46 C20H23N3O2

δH 8.63 (1H, s), 8.12 (1H, d, J = 8.6 Hz), 8.01 (1H, ovl), 8.00 (1H, ovl), 7.94 (1H, d, J = 7.4Hz), 7.60 (1H, ovl), 7.59 (1H, ovl), 4.06 (1H, dd, J = 7.2, 4.3Hz), 3, 59 (2H, t, J = 6.1Hz), 3.13 (1H, m), 2.57 (2H, m), 2.46 (1H, m), 1.99 (1H, m), 1.87 (2H, m), 1.73 (2H, m), 1.72 (2H, m), 1.56 (1H, m);

δC 181.7, 169.4, 136.2, 134.5, 129.9, 129.8, 128.9 (2C), 128.8, 128.0, 125.1, 124.6, 61, 6, 59.9, 54.6, 51.7, 31.8, 30.1, 26.3, 23.1.

Example 3F. (S) -3- (naphthalen-2-yl) -5- (1-pentylpiperidin-2-yl) -1,2,4-oxadiazole (OXA37)

OXA37 was purified by HPLC on Luna Synergi Fusion-RP (4 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (85:15) as eluent (flow rate 1 mL / min, tr = 31.1 min).

OXA37 C22H27N3O

δH 8.67 (1H, s), 8.14 (1H, d, J = 8.5 Hz), 8.02 (1H, d, J = 8.7 Hz), 7.98 (1H, ovl) , 7.95 (1H, d, J = 7.4 Hz), 7.62 (1H, ovl), 7.60 (1H, ovl), 4.04 (1H, br t, J = 6.2 Hz ), 3.12 (1H, m), 2.43 (2H, m), 2.34 (1H, m), 2.00 (2H, m), 1.88 (1H, m), 1.75 (2H, m), 1.55 (3H, m), 1.27 (4H, m), 0.87 (3H, t, J = 6.9 Hz);

δC 182.3, 169.5, 136.1, 134.5, 129.9, 129.8, 128.9 (2C), 128.7 (2C), 128.0, 124.6, 61.4 , 55.9, 54.6, 31.3, 30.6, 29.2, 24.2, 22.3, 14.3.

Example 4. GENERAL PROCEDURE FOR N-ALKYLATION ON OXA20 The alkylation on OXA20 with the desired methyl bromide ester Br (CH2) nCOOMe or the desired alkyl bromide was carried out in the same operating condition described in steps a) and d) of the Example 2. Hydrolysis and reduction at the methyl ester functional group were carried out in the same operating condition described in steps b) and c) of Example 2.

a) acyl bromide, DIPEA in CH3CN, 60 ° C; b) LiOH in THF / H, 0 ° C and therefore the ambient temperature; c) DIBAL-H in THF, 0 ° C; d) alkyl bromide, DIPEA in CH3CN, 60 ° C. Example 4A: (S) -methyl 4- (2- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) pyrrolidin-1-yl) butanoate (OXA24)

OXA24 was purified by HPLC on Luna Synergi Fusion-RP (4 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (75:15) as eluent (flow rate 1 mL / min, tR = 19.9 min).

OXA24 C21H23N3O3

δH 8.63 (1H, s), 8.12 (1H, d, J = 8.5 Hz), 8.00 (1H, ovl), 7.99 (1H, ovl), 7.94 (1H, d, J = 6.9 Hz), 7.59 (1H, ovl), 7.58 (1H, ovl), 4.06 (1H, dd, J = 8.5, 5.8 Hz), 3, 56 (3H, s), 3.26 (1H, m), 2.77 (2H, m), 2.57 (2H, m), 2.39 (2H, m), 2.20 (1H, m ), 2.09 (1H, m), 2.00 (1H, m), 1.79 (2H, m);

δC 182.3, 175.5, 169.5, 136.2, 134.6, 130.0, 129.9, 129.0 (2C), 128.8, 128.1, 125.3, 124, 7, 61.6, 55.0, 54.7, 52.1, 32.3, 31.4, 24.7, 24.4.

Example 4B (S) -4- (2- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) pyrrolidin-1-yl) butanoic (OXA40)

OXA40 was purified by HPLC on Luna Synergi Fusion-RP (4 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (7: 3) as eluent (flow rate 1 mL / min, tR = 12 min) ;

OXA40 C20H21N3O3

δH 8.63 (1H, s), 8.11 (1H, d, J = 8.5 Hz), 8.01 (1H, ovl), 7.99 (1H, ovl), 7.93 (1H, d, J = 7.6Hz), 7.59 (1H, ovl), 7.58 (1H, ovl), 4.12 (1H, dd, J = 8.4, 6.0Hz), 3, 28 (1H, ovl), 2.79 (1H, m), 2.61 (1H, ovl), 2.58 (1H, ovl), 2.37 (1H, ovl), 2.33 (2H, ovl ), 2.19 (1H, m), 2.10 (1H, m), 2.02 (1H, m), 181 (2H, m);

δC 182.3, 178.0, 169.5, 136.1, 134.5, 129.9, 129.8, 129.0, 128.9, 128.8, 128.0, 125.3, 124 , 6, 61.5, 55.2, 54.4, 33.2, 31.4, 24.9, 24.3.

Example 4C. (S) -4- (2- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) pyrrolidin-1-yl) butan-1-ol (OXA42)

OXA42 was purified by HPLC on Luna Synergi Fusion-RP (4 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (75:25) as eluent (flow rate 1 mL / min, tR = 14.2 min).

OXA42 C20H23N3O2

δH 8.63 (1H, s), 8.11 (1H, d, J = 8.5 Hz), 8.01 (1H, dovl), 7.99 (1H, ovl), 7.95 (1H, d, J = 7.1Hz), 7.61 (1H, ovl), 7.60 (1H, ovl), 4.08 (1H, dd, J = 8.3, 6.1Hz), 3, 53 (2H, t, J = 5.9 Hz), 3.27 (1H, ovl), 2.78 (1H, m), 2.59 (1H, m), 2.54 (1H, m), 2.38 (1H, m), 2.21 (1H, m), 2.12 (1H, m), 2.02 (1H, m), 160 (2H, ovl), 1.57 (2H, ovl );

δC 182.3, 169.5, 136.1, 134.4, 130.0, 129.8, 129.1, 128.9, 128.7, 128.0, 125.4, 124.6, 62 , 7, 61.4, 55.8, 54.5, 31.5, 31.3, 26.1, 24.2.

Example 4D. (S) -methyl 3- (2- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) pyrrolidine-1-yl) propanoate (OXA44) OXA44 was purified by HPLC on Luna Synergi Fusion-RP (4µm; 4.6 mm ID x 250 mm) with MeOH / H2O (80:20) as eluent (flow rate 1 mL / min, tR = 7.0 min).

OXA44 C20H21N3O3

δH 8.62 (1H, s), 8.10 (1H, d, J = 8.5 Hz), 8.00 (1H, ovl), 7.99 (1H, ovl), 7.96 (1H, d, J = 7.1Hz), 7.61 (1H, ovl), 7.60 (1H, ovl), 4.12 (1H, dd, J = 8.3, 5.4Hz), 3.61 ( 3H, s), 3.24 (1H, m), 3.08 (1H, m), 2.83 (1H, m), 2.62 (1H, m), 2.54 (2H, m), 2.38 (1H, m), 2.21 (1H, m), 2.08 (1H, m), 2.01 (1H, m);

δC 182.4, 174.3, 169.5, 136.2, 134.5, 129.9, 129.8, 128.9, 128.8, 128.7, 128.0, 125.3, 124.6, 61.2, 54 , 5, 52.2, 50.9, 34.4, 31.5, 24.5.

Example 4E. (S) -3- (2- (3- (naphthalen-2-yl) -1,2,4-oxadiazol-5-yl) pyrrolidin-1-yl) propan-1-ol (OXA47)

OXA47 was purified by HPLC on Luna Omega Polar C18 (5 μm; 4.6 mm ID x 250 mm) with MeOH / H2O (68:32) as eluent (flow rate 1 mL / min, tR = 27.0 min ).

OXA47 C19H21N3O2

δH 8.63 (1H, s), 8.11 (1H, d, J = 8.5 Hz), 8.01 (1H, d, ovl), 7.99 (1H, ovl), 7.95 ( 1H, d, J = 7.1Hz), 7.61 (1H, ovl), 7.60 (1H, ovl), 4.08 (1H, dd, J = 8.5, 5.8Hz), 3, 63 (2H, t, J = 6.1Hz), 3.28 (1H, ovl), 2.88 (1H, m), 2.62 (1H, m), 2.59 (1H, m), 2.40 (1H, m), 2.20 (1H, m), 2.11 (1H, m), 2.02 (1H, m), 1.76 (2H, ovl);

δC 182.3, 169.5, 136.4, 134.5, 129.9, 129.8, 129.0, 128.9, 128.8, 128.0, 125.4, 124.6, 61 , 6, 61.5, 59.2, 54.3, 32.2, 31.4, 24.3.

Example 4F. (S) -3- (naphthalen-2-yl) -5- (1-pentylpyrrolidin-2-yl) -1,2,4-oxadiazole (OXA38)

OXA38 was purified by HPLC on Luna Synergi Fusion-RP (4 µm; 4.6 mm ID x 250 mm) with MeOH / H2O (88:12) as eluent (flow rate 1 mL / min, tR = 13.6 min).

OXA38 C21H25N3O

δH 8.62 (1H, s), 8.11 (1H, d, J = 8.6 Hz), 8.00 (1H, ovl), 7.99 (1H, ovl), 7.93 (1H, d, J = 7.2Hz), 7.59 (1H, ovl), 7.58 (1H, ovl), 4.04 (1H, dd, J = 8.4, 6.1Hz), 3, 24 (1H, m), 2.72 (1H, m), 2.56 (1H, dd, J = 17.0, 8.5 Hz), 2.48 (1H, m), 2.37 (1H , m), 2.20 (1H, m), 2.10 (1H, m), 2.00 (1H, m), 1.54 (2H, m), 1.28 (4H, m), 0 86 (3H, t, J = 6.2 Hz);

δC 182.3, 169.5, 136.1, 134.5, 129.9, 129.8, 128.9 (2C), 128.7 (2C), 128.0, 124.6, 61.4 , 55.9, 54.6, 31.3, 30.6, 29.2, 24.2, 22.3, 14.3.

EXAMPLE 5 BIOLOGICAL ACTIVITY

The biological activity of the selected compounds (Table 1) was tested in vitro using a cell model transfected with reporter genes, on the FXR receptor in comparison with the control agonist, chenodeoxycholic acid (CDCA), a primary bile acid that acts as an endogenous receptor ligand. HepG2 cells were cultured at 37 ° C in E-MEM ("Earl's salt Minimum Essential Medium") with the addition of 10% fetal bovine serum (FBS), L- 1% glutamine and 1% penicillin / streptomycin. Transfection experiments were performed using Fugene HD (Promega) reagent according to the manufacturer's instructions. Cells were plated in 24-well plates at 5 × 10 <4> cells / well.

HepG2 cells were transfected with 100 ng of pSG5-FXR vector, 100 ng of pSG5-RXR vector, 100 ng of pGL4.70 Renilla vector, a plasmid encoding the human Renilla gene, and 200 ng of p reporter vector (hsp27) -TK-LUC containing the reactive FXR element IR1 cloned from the promoter of the heat shock protein 27 (hsp27).

24 hours after transfection, cells were stimulated for 18 hours with the test compounds alone and in the presence of 10 µM CDCA.

After the treatments, the cells were lysed in 100 μl of lysis buffer (25 mM Tris-phosphate, pH 7.8, 2 mM DTT, 10% glycerol, 1% Triton X-100); 10 μl of cell lysate from each sample were assayed for luciferase activity using a Dual Luciferase Reporter Assay System (Promega Italia S.r.l., Milan, Italy) according to the manufacturer's instructions. Luminescence was measured using Glomax 20/20 luminometer (Promega Italia S.r.l., Milan, Italy). Luciferase activity (Luciferase Recording Unit, RLU) was normalized using Renilla activity (Renilla Recording Unit, RRU). The IC50 values of each compound were determined by constructing a dose-response curve in HepG2 cells transfected as described above and then treated with increasing concentrations of compounds (0.1-100 µm) in the presence of 10 µM CDCA.

Table 1 reports the effectiveness of the selected compounds included in Formula I as a percentage of antagonistic activity compared to that of the CDCA for which the transactivation activity was considered equal to 100%. Each compound was tested at a concentration of 50 μM.

None of the compounds belonging to formula I showed agonist activity on FXR.

None of the compounds belonging to formula I showed agonist / antagonist activity on GPBAR1.

For GPBAR1-mediated transactivation, HEK-293T cells were transfected with 200 ng of the plasmid pGL4.29 (Promega), a reporter vector containing the cAMP response element (CRE) cloned upstream of the luciferase luciferase reporter gene luc2P, 100 ng of the human pCMVSport6-GPBAR1 vector, and 100 ng of the pGL4.70 Renilla vector, a plasmid encoding the human Renilla gene. In control experiments, HEK-293T cells were transfected with pGL4.29 (Promega) and Renilla pGL4.70 vectors only, to rule out any possibility that the compounds could activate CRE independently of GPBAR1.

Table 1

Preferred examples included in the general formula Ib are OXA7 and OXA17 with an efficacy of 80% and 95% in antagonizing the transactivation of CDCA and the IC50 values of 0.58 μm and 1.17 μm, respectively.

Preferred examples included in the general Ic formula are OXA21, OXA28 and OXA41 with an efficacy of 96%, 85% and 82% in antagonizing the transactivation of CDCA and the IC50 values of 0.127 μM, 0.067 and 7 μm.

Claims (1)

CLAIMS 1.- Compound of formula (I): where A1 and A2 are selected from the group consisting of CH2 and NR1 provided that A1 and A2 are not at the same time CH2 or NR1, R1 is selected by CH3 and (CH2) 2-4-R2, R2 is selected from the group consisting of H, CH3, COOR3 and CH2OH, R3 is selected from the group consisting of H and C1-4 alkyl, R4, R5, R6, R7, R8, R9 and R10 are independently selected from the group consisting of H, C1-4 alkyl, O-C1-4 alkyl, halogen and OH. n is selected from 1 and 2 or pharmaceutically acceptable salts and solvates. 2.- Compound according to claim 1, wherein n is 1. 3.- Compound according to claim 2 having the formula (Ia) where R1 is selected by CH3 and (CH2) 2-4-R2, R2 is selected from the group consisting of H, CH3, COOR3 and CH2OH, R3 is selected from the group consisting of H and C1-4 alkyl, R4, R5, R6, R7, R8, R9 and R10 are independently selected from the group consisting of H, C1-4 alkyl, O-C1-4 alkyl, halogen and OH. 4.- Compound according to claim 1, wherein n is 2. 5.- Compound according to claim 4 having the formula Ib where R1 is selected by CH3 and (CH2) 2-4-R2, R2 is selected from the group consisting of H, CH3, COOR3 and CH2OH, R3 is selected from the group consisting of H and C1-4 alkyl, R4, R5, R6, R7, R8, R9 and R10 are independently selected from the group consisting of H, C1-4 alkyl, O-C1-4 alkyl, halogen and OH. 6.- Compound according to claim 4 having the formula Ic where R1 is selected by CH3 and (CH2) 2-4-R2, R2 is selected from the group consisting of H, CH3, COOR3 and CH2OH, R3 is selected from the group consisting of H and C1-4 alkyl, R4, R5, R6, R7, R8, R9 and R10 are independently selected from the group consisting of H, C1-4 alkyl, O-C1-4 alkyl, halogen and OH. 7. A compound according to any one of claims 1 to 6, wherein the compound is selected from the group consisting of 8.- Pharmaceutical composition comprising a compound of Formula (I): where A1 and A2 are selected from the group consisting of CH2 and NR1 provided that A1 and A2 are not at the same time CH2 or NR1, R1 is selected by H, CH3 and CH2) 2-4-R2, R2 is selected from the group consisting of H, CH3, COOR3 and CH2OH, R3 is selected from the group consisting of H and C1-4 alkyl, R4, R5, R6, R7, R8, R9 and R10 are independently selected from the group consisting of H, C1-4 alkyl, O-C1-4 alkyl, halogen and OH. n is selected between 1 and 2 and at least one pharmaceutically acceptable excipient. 9.- Compound of Formula (I): where A1 and A2 are selected from the group consisting of CH2 and NR1 provided that A1 and A2 are not at the same time CH2 or NR1, R1 is selected from H, CH3 and (CH2) 2-4-R2, R2 is selected from the group consisting of H, CH3, COOR3 and CH2OH, R3 is selected from the group consisting of H and C1-4 alkyl, R4, R5, R6, R7, R8, R9 and R10 are independently selected from the group consisting of H, C1-4 alkyl, O-C1-4 alkyl, halogen and OH. n is selected between 1 and 2 for use as a drug. 10.- Compounds according to claim 9 for use in the prevention and / or treatment of a disorder selected from the group consisting of gastrointestinal disorders, liver disorders, cardiovascular disorders, vascular disorders, pulmonary disorders, metabolic diseases, infectious diseases, cancer, disorders kidney, inflammatory disorders including immune-mediated and neurological disorders. 11.- Compounds according to claim 10 for use according to claim 10, characterized in that said disorder is selected from the group consisting of systemic lupus erythematosus, rheumatoid arthritis, Sjogren's syndrome, scleroderma, spondyloarthritis, vasculitis, sarcoidosis, Mediterranean fever, polymyositis and dermatomyositis, Behcet's syndrome, acquired immune deficiency syndrome and associated disorders, virus B and virus C infections, Alzheimer's disease and other dementias, Parkinson's disease and other movement disorders, amyotrophic lateral sclerosis and other neuromotor disorders, multiple sclerosis and other demyelinating diseases, myasthenia and muscular dystrophies, primary biliary cirrhosis, cerebrotendinous xanthomatosis, primary sclerosing cholangitis, drug-induced cholestasis, intrahepatic cholestasis of pregnancy, cholestasis associated with parenteral nutrition, cholestasis associated with bacterial proliferation or sepsis, autoimmune hepatitis, hepatitis chronic, ep alcoholic atopathy and non-alcoholic fatty liver disease, non-alcoholic steatohepatitis, liver transplant, congenital hepatic fibrosis, granulomatous liver disease, intra- or extra-hepatic malignancy, Wilson's disease, hemochromatosis, alpha 1-antitrypsin deficiency, inflammatory bowel disease, disease Crohn's disease, ulcerative colitis, indeterminate colitis, irritable bowel syndrome, bacterial overgrowth, acute and chronic pancreatitis, malabsorption, post-radiation colitis, microscopic colitis, diabetic nephropathy, hypertensive nephropathy, chronic glomerulonephritis, chronic graft glomerulonephritis, tubulointerstitial disease chronic, renal vascular disorders, atherosclerosis, arteriosclerosis, dyslipidemia, hypercholesterolemia, hypertriglyceridemia, arterial hypertension, inflammatory cardiac disorders, myocarditis, endocarditis, cardiac ischaemia, stable angina, unstable angina, myocardial infarction, cerebrovascular disorders, pulmonary hypertension, stroke peripheral arterial disease, occlusive peripheral arterial disease, peripheral arterial disease, asthma, cystic fibrosis, obstructive respiratory disease, interstitial lung disease, primary or secondary pulmonary fibrosis, insulin resistance, metabolic syndrome, type I and type II diabetes, hypoglycemia, adrenal cortex disorders, adrenal cortex impairment, obesity, conditions associated with bariatric surgery, liver cancer, types of bile duct cancer, esophageal cancer, pancreatic cancer, gastric cancer, colorectal cancer, breast cancer, ovarian cancer and associated conditions resistance to chemotherapy.