Thienopyrimidylamine derivatives as phgdh inhibitors

Abstract

The present invention generally relates to compounds of formula (I) inhibiting 3- phosphoglycerate dehydrogenase (PHGDH) activity; particularly, the invention relates to compounds that are thienopyrimidylamine derivatives, including pharmaceutically acceptable salts thereof, methods of preparing such compounds, and therapeutic use thereof. The compounds of the invention may be useful for instance in the treatment of many disorders associated with PHGDH, for instance in the treatment of many disorders associated with fibrosis, such as idiopathic pulmonary fibrosis (IPF).

Description

[0001] THIENOPYRIMIDYL AMINE DERIVATIVES AS PHGDH INHIBITORS

[0002] FIELD OF THE INVENTION

[0003] The present invention generally relates to compounds inhibiting 3 -phosphoglycerate dehydrogenase (hereinafter PHGDH) receptor; the invention relates to compounds that are thienopyrimidylamine derivatives, including pharmaceutically acceptable salts thereof, methods of preparing such compounds, and therapeutic use thereof.

[0004] The compounds of the invention may be useful for instance in the treatment of many disorders associated with fibrosis, such as idiopathic pulmonary fibrosis (IPF).

[0005] BACKGROUND OF THE INVENTION

[0006] 3 -phosphoglycerate dehydrogenase (PHGDH) is the enzyme that catalyzes the first ratelimiting step in the de novo biosynthesis of L-serine starting from glucose, which is the conversion of 3 -phosphoglycerate into 3-phosphohydroxypyruvate with a reduction of nicotinamide adenine dinucleotide (NAD+) to NADH, and it is considered as a major enzyme in the diversion of glycolysis towards serine synthesis. Serine, a critical amino acid for protein and nucleic acid biosynthesis, in turn is metabolized and incorporated into a variety of biomolecules including glycine (see YangM., Vousden, KH, Nature Reviews Cancer 2016,16, 650 662).

[0007] PHGDH is required to promote collagen protein synthesis. Indeed, glycine accounts for one- third of all amino acids within the collagen molecule, and the high content of glycine is critical for the stabilization of collagen helix.

[0008] Collagen is the main structural protein in the extracellular space, and it is produced in excess in IPF patients during fibroblasts into myofibroblasts differentiation that accumulates within the fibrotic tissue and leads to loss of organ architecture and function.

[0009] Moreover, PHGDH and the serine / glycine synthesis pathway is part of a wider network which links glycolysis with one-carbon metabolism and nucleotide synthesis contributing to cell proliferation in pathologies such as inflammation and fibrosis.

[0010] Therefore, the PHGDH inhibition has the potential to reduce the aberrant production and release of collagen such as inhibit cell proliferation (see Selvarajah et al., Science Signaling, 2019;12(582):eaav 3048).

[0011] Various compounds have been described in the literature as PHGDH inhibitors.

[0012] Raze Therapeutics disclosed in various patent applications (for example WO 2017156165 or WO 2017156179) compounds which are effective as orthosteric PHGDH inhibitors, and their use in the treatment of many PHGDH-mediated disorders, in particular melanoma, breast, or lung cancer.

[0013] Boehringer Ingelheim disclosed in patent application WO 2018167019 tosylacetate based compounds and derivatives which are effective as PHGDH inhibitors and their use in the treatment of PHGDH-mediated disorders, such as treatment and / or prevention of cancer, infections, inflammations or autoimmune diseases.

[0014] Alternative PHGDH inhibitors, promising for the prevention and / or treatment of fibrosis, were described as noncompetitive inhibitors or allosteric inhibitors of PHGDH enzyme (see Pacold et al, Nat Chem Biol. 2016; 12: 452-458).

[0015] Examples of said PHGDH inhibitors useful for treating fibrotic diseases are disclosed in WO2016115463 (Whitehead Institute For Biomedical Research; Dana-Farber Cancer Institute, Inc).

[0016] Fibrosis is a pathological wound healing in which connective tissue replaces normal parenchymal tissue to the extent that it goes unchecked, leading to considerable tissue remodeling and the formation of permanent scar tissue (see Wynn, Thomas A., 2004 Nature Reviews 4 (8): 583-594).

[0017] Fibrosis can occur in many tissues within the body, typically as a result of inflammation or damage, and examples include liver, lungs, kidney, brain and heart.

[0018] Idiopathic pulmonary fibrosis (IPF) is a rare, progressive illness of the respiratory system, characterized by the thickening and stiffening of lung tissue, associated with the formation of scar tissue. It is a type of chronic scarring lung disease characterized by a progressive and irreversible decline in lung function, (see HR, Egan JJ, et al. American Journal of Respiratory and Critical Care Medicine, 2011, 183 (6): 788-824).

[0019] About 3 million people are affected globally and IPF seems to become more common over the year, the incidence is expected to double by 2030 (see Fernandex Perez ER, et al, Chest, 2020; 137:129-137). Mortality is comparable to lung cancer, the median survival is 2-3 years after diagnosis, and respiratory failure is the most common cause of death (see Strongman H, et a., Adv Ther. 2018;35: 724-736).

[0020] The currently two FDA-approved drugs on the market, Nintedanib and Pirfenidone, slow the progression of the disease and prolong the life expectancy of IPF patients; however, the treated patients continue to experience lung function loss and premature death and the adverse events of both Nintedanib and Pirfenidone critically affect the patient’s quality of life.

[0021] The IPF is therefore a high unmet medical need and many efforts have been done in the recent past years to develop alternative treatments.

[0022] Despite the above prior art, there remains a potential for developing medicaments comprising a PHGDH inhibitor useful for the prevention and / or treatment of fibrosis, in particular IPF.

[0023] The underlying problem of the present invention therefore lies in the provision of a medicament comprising a compound of formula (I) for the prevention and / or treatment of fibrosis, in particular IPF.

[0024] The state of the art does not describe or suggest the thienopyrimidylamine compounds of general formula (I) of the present invention which may represent a solution to the afore mentioned unmet medical need.

[0025] SUMMARY OF THE INVENTION

[0026] In a first aspect the invention refers to a compound of formula (I) wherein at least one of Xi and X2 is S, and the other one is CH;

[0027] Ri is H or selected from the group consisting of -(Ci-Ce)alkyl, -(Ci-C6)aminoalkyl, -ORs, - SRs, heteroaryl and -(C3-C7)heterocycloalkyl, wherein said -(Ci-C6)aminoalkyl, heteroaryl, -(C3- C7)heterocycloalkyl are optionally substituted by one or more group selected from -(Ci-Ce)alkyl or -ORs;

[0028] R2 and R3 are independently H or -(Ci-Ce)alkyl, optionally substituted by -ORs, or are fused together in a -(C3-C7)heterocycloalkyl;

[0029] R4 and R5 are independently H or -(Ci-Ce)alkyl;

[0030] A is a ring selected from the group consisting of aryl and heteroaryl, wherein any of such aryl, heteroaryl are optionally substituted by one or more groups selected from halogen, -(Ci- C6)alkyl, -ORs, -NH(CO)-(Ci-C6)alkyl, -(CO)-NR4R5;

[0031] LI is a bond or selected from the group consisting of -(Ci-Ce)alkylene-, -SO2-(Ci- Ce)alkylene- and -(C0)-NReR7-, wherein when Li is -(C0)-NReR7- and Re and R7 are fused together in a -(C3-C7)heterocycloalkyl, the carboxyl group -(C0)-0-R4 of formula (I) is linked to anyone of the C atom of such -(C3-C7)heterocycloalkyl; and pharmaceutically acceptable salts thereof.

[0032] 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.

[0033] In a third aspect, the invention refers to a compound of formula (I) for use as a medicament.

[0034] In a further aspect, the invention refers to a compound of formula (I) for use in treating diseases, disorders or conditions associated with dysregulation of 3 -phosphoglycerate dehydrogenase (PHGDH).

[0035] 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.

[0036] 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)

[0037] DETAILED DESCRIPTION OF THE INVENTION

[0038] Unless otherwise provided, the term “compound of formula (I)” comprises in its meaning solvates, stereoisomers, tautomers, deuterated and pharmaceutically acceptable salts or solvates thereof.

[0039] 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.

[0040] 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.

[0041] 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.

[0042] 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, iodic acid, formic acid, benzoic acid, sulfuric acid, nitric acid, phosphoric acid, methane sulfonic acid, camphor sulfonic acid, nitric acid, acetic acid, oxalic acid, maleic acid, fumaric acid, succinic acid, p-toluenesulfonic acid, trifluoroacetic acid, 2-naphthoic acid, tartaric acid, l-hydroxy-2- naphthoic acid, naphthalene-2,7-disulfonic acid and citric acid.

[0043] 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 might be isolated by crystallization, 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 non-stoichiometric amount of the solvent molecules.

[0044] The term "stereoisomer" refers to isomers of identical constitution that differ in the arrangement of their atoms in space. Enantiomers and diastereomers are examples of stereoisomers.

[0045] The term "enantiomer" refers to one of a pair of molecular species that are mirror images of each other and are not superimposable.

[0046] The term "diastereomer " refers to stereoisomers that are not mirror images.

[0047] 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.

[0048] When one of the compounds of this invention is defined as a specific enantiomer or diastereoisomer, the number reported in the name of this specific enantiomer or diastereoisomer is intended to define the order of elution of the compound during any process of chiral separation. For example, a compound defined as “Enantiomer 1” is intended to be the first eluted compound in the above described chiral separation process.

[0049] In case of separation of 4 diastereoisomers with two different chromatographic methods, a compound defined as “Diastereoisomer 1” is intended to be the first eluted compound in the first chiral separation mentioned, while a compound defined as “Diastereoisomer 3” is intended to be the first eluted compound in the second chiral separation mentioned.

[0050] The symbols "R" and "S" represent the configuration of substituents around a chiral carbon atom(s) and are intended to be used as defined in the literature (TUPAC Recommendations 1996, Pure and Applied Chemistry, 68:2193-2222 (1996)).

[0051] 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.

[0052] The term "deuterium" refers to the isotopic deuterium of hydrogen (H).

[0053] The term "deuterated" refers to the case where the hydrogen atoms on an alkyl, cycloalkyl, aryl, heteroaryl group are substituted by at least one isotopic deuterium, with the upper limit of the number of deuterium substituents being equal to the sum of the number of hydrogen atoms that can be substituted. Unless otherwise indicated, the number of deuterium substituents is any integer between 1 and said upper limit, preferably substitution by 1 to 20 deuterium atoms, more preferably 1 to 10 deuterium atoms, more preferably 1 to 6 deuterium atoms, and further preferably 1 to 3 deuterium atoms.

[0054] The term “halogen” or “halogen atoms” or “halo” as used herein includes fluorine, chlorine, bromine and iodine atom.

[0055] 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 1 and y is 6, for example, the term includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, t-butyl, n-pentyl and n- hexyl.

[0056] The term “(Cx-Cy)haloalkyl” wherein x and y are integers, refers to the above defined “(Cx- Cy)alkyl” groups wherein one or more hydrogen atoms are replaced by one or more halogen atoms, which can be the same or different. Examples of said “(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.

[0057] The term "(Cx-Cy)aminoalkyl” wherein x and y are integers, refers to the above defined “(Cx- Cy)alkyl” groups wherein one or more hydrogen atoms are replaced by one or more amino group, which can be the same or different. Examples of "(Cx-Cy)aminoalkyl” include, for instance, aminomethyl.

[0058] The term "(Cx-Cy)alkylene" wherein x and y are integers, refers to a straight or branched “(Cx-Cy)alkyl” radical having in total two unsatisfied valences. Example of such "(Cx-Cy)alkylene" groups may include for example a divalent methylene radical and -CH(CH3)-.

[0059] The term “partially unsaturated” refers to a ring moiety that includes at least one double or triple bond. The term “partially unsaturated” is intended to encompass rings having multiple sites of unsaturation but is not intended to include aryl or heteroaryl moieties, as herein defined.

[0060] The term “(Cx-Cy)cycloalkyl” wherein x and y are integers, refers to saturated or partially unsatured mono- or poly-cyclic hydrocarbon groups containing the indicated number of ring carbon atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl.

[0061] The term “(Cx-Cy)heterocycloalkyl” wherein x and y are integers, refers to saturated or partially unsaturated monocyclic or polycyclic groups containing the indicated number of ring carbon atoms in which at least one ring carbon atom is replaced by at least one heteroatom or hetero-group (e.g. N, NH, S or O) or may bear an oxo (=0) substituent group. The said heterocycloalkyl (i.e. heterocyclic radical or group) might be further optionally substituted on the available positions in the ring, namely on a carbon atom, or on a heteroatom or hetero-group available for substitution. Substitution on a carbon atom includes spiro di-substitution as well as substitution on two adjacent carbon atoms, in both cases thus forming an additional condensed 5- to 6-membered heterocyclic ring. Non limiting examples of “(Cx-Cy)heterocycloalkyl” are represented by pyrrolidinyl, piperazinyl, piperidinyl, morpholinyl, azetidinyl, thiomorpholinyl, pyrrolinyl, dihydro- or tetrahydro-thiazolyl, oxetanyl, tetrahydropyranyl, pyranyl, dihydro- or tetrahy dro-furanyl .

[0062] The term “aryl” refers to mono- or bi-cyclic carbon ring systems wherein the ring is aromatic. Examples of suitable aryl ring systems include, for instance, phenyl or naphthyl.

[0063] The term “heteroaryl” refers to mono- or bi- or tri-cyclic ring systems with 5 to 20, preferably from 5 to 15 ring atoms, in which at least one ring is aromatic and in which at least one ring atom is a heteroatom (e.g. N, NH, S or O). Examples of “heteroaryl” include thiophenyl, pyrrolyl, pyrazolyl, imidazolyl, isoxazolyl, oxazolyl, isothiazolyl, thiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, benzofuranyl, dihydrobenzofuranyl, triazinyl.

[0064] A group may be optionally substituted, wherein the term “optionally substituted” refers to being substituted or unsubstituted. When the term "one or more " refers to any atoms or groups as substituents of the groups of the compound of formula (I), it is intended that from 1 to 3, preferably 1 to 2, more preferably 1 of such substituents may replace hydrogens on such variables.

[0065] A bond pointing to a wavy or squiggly line, such as 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.

[0066] The term "bond" used to define a substituent refers to the situation where the two functional groups which the substituent is connected to are directly linked to each other with no additional atoms in between.

[0067] The carbonyl group is herein preferably represented as -(CO)- as an alternative to the other common representations such as -CO-, C=O, -C(O)- or -C(=O)-.

[0068] A dash (“-”) that is not between two letters or symbols is meant to represent the point of attachment for a substituent.

[0069] The term “ICso” refers to the half maximal inhibitory concentration as a measure of the potency of a substance in inhibiting a specific biological or biochemical function.

[0070] The term “pICso” refers to the negative logarithm of the ICso value expressed as molar concentration.

[0071] The term “PHGDH” or “phosphoglycerate dehydrogenase”, is alternatively referred to as 3- PGDH, 3PGDHm HEL-S-113, NLS, PDG, PGAD, PGD, PGDH, PHGDHD, or SERA. The term “PHGDH” encompasses mutants, variants, homologs, fragments, and synthetically modified phosphoglycerate dehydrogenases.

[0072] Whenever basic amino or quaternary ammonium groups are present in the compounds of formula (I), pharmaceutically 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, emipamoate, xinafoate and naphthalene disulfonate.

[0073] Likewise, in the presence of acidic groups, such as for instance COOH groups, corresponding pharmaceutically acceptable cations may be present, for instance including alkaline or alkaline earth metal ions, including sodium, potassium, calcium and the like.

[0074] As above indicated, the present invention refers to a series of compounds represented by the general formula (I), as herein below described in detail, which are endowed with an inhibitory activity versus 3 -phosphoglycerate dehydrogenase (hereinafter defined as PHGDH) receptor.

[0075] The compounds of formula (I) of the present invention are able to act as inhibitors of PHGDH receptor in a substantive and effective way, particularly appreciated by the skilled person when looking at a suitable and efficacious compound useful for the treatment of many disorders associated with fibrosis, and in particular the treatment of idiopathic pulmonary fibrosis (IPF).

[0076] As indicated in the experimental part, the compounds of formula (I) of the invention have an activity as shown in Table 29, wherein for each compound tested in the Low NAD biochemical Assay is reported the potency expressed as pIC50.

[0077] As it can be appreciated, all the compounds of the present invention according to Table 29, show a potency with respect to their inhibitory activity on receptor PHGDH higher than 5.5.

[0078] For some representative compounds of formula (I) pICso values were determined also in high NAD conditions assay showing a potency with respect to their inhibitory activity on PHGDH receptor, expressed as pICso values, higher than 4.5.

[0079] Some representative compounds of formula (I) were tested also in a cellular serine flux assay based on13C3-Serine MS read out, using breast cancer cells lines (MDA-MB468). PHGDH is a key enzyme for Serine biosynthesis, and it is described that breast cancer cells (MDA-MB468) can be dependent on PHGDH for their growth and survival. In the cellular assay mentioned above representative compounds tested demonstrated good inhibitory activity of the PHGDH receptor.

[0080] Accordingly, the compounds of formula (I) can be used in the treatment of fibrosis, and in particular treatment of idiopathic pulmonary fibrosis, whenever PHGDH receptors are involved.

[0081] Thus, in one aspect the present invention relates to a compound of general formula (I) wherein at least one of Xi and X2 is S, and the other one is CH;

[0082] Ri is H or selected from the group consisting of -(Ci-Ce)alkyl, -(Ci-C6)aminoalkyl, -ORs, - SRs, heteroaryl and -(C3-C7)heterocycloalkyl, wherein said -(Ci-C6)aminoalkyl, heteroaryl, -(C3- C7)heterocycloalkyl are optionally substituted by one or more group selected from -(Ci-Ce)alkyl or -ORs;

[0083] R2 and Rj are independently H or -(Ci-Ce)alkyl, optionally substituted by -ORs, or are fused together in a -(C3-C7)heterocycloalkyl;

[0084] R4 and R5 are independently H or -(Ci-Ce)alkyl; A is a ring selected from the group consisting of aryl and heteroaryl, wherein any of such aryl, heteroaryl are optionally substituted by one or more groups selected from halogen, -(Ci- C6)alkyl, -ORs, -NH(CO)-(Ci-C6)alkyl, -(CO)-NR4Rs;

[0085] Li is a bond or selected from the group consisting of -(Ci-Ce)alkylene-, -SO2-(Ci- Ce)alkylene- and -(CO)-NReR7-, wherein when Li is -(CO)-NReR7- and Re and R? are fused together in a -(C3-C7)heterocycloalkyl, the carboxyl group -(CO)-O-R4 of formula (I) is linked to anyone of the C atom of such -(C3-C7)heterocycloalkyl; and pharmaceutically acceptable salts thereof.

[0086] All the listed groups for each of the variable moieties Xi, X2, Ri, Rz, Rj, Rt, Rs, Re, R7, Li and A of the compounds of the invention have to be intended as alternatives and may be combined with each other in embodiments which are included in the scope of the invention.

[0087] The 5-membered ring comprising Xi and X2 is a thiophene ring, that is fused to a pyrimidine to give an aromatic bicyclic ring selected from thieno[2,3]pyrimidine or thieno[3,2]pyrimidine, which has a substituted amine group on the pyrimidine ring.

[0088] In one embodiment Xi is S and X2 is CH. In another embodiment, X2 is S and Xi is CH.

[0089] In one embodiment, Ri is H or selected from the group consisting of -(Ci-Ce)alkyl, -(Ci- Ce)aminoalkyl, -ORs, -SRs, heteroaryl and -(C3-C7)heterocycloalkyl, wherein said -(Ci- Ce)aminoalkyl, heteroaryl, -(C3-C7)heterocycloalkyl are optionally substituted by one or more group selected from -(Ci-Ce)alkyl or -ORs.

[0090] In a preferred embodiment, Ri is H or selected from the group consisting of -(Ci-Ce)alkyl, - ORs and -SRs. In a more preferred embodiment, Ri is H or -(Ci-Ce)alkyl. In a more preferred embodiment, Ri is H or -(Ci-C4)alkyl. In another preferred embodiment, Ri is -(Ci-C6)aminoalkyl, optionally substituted by one or more group selected from -(Ci-C4)alkyl or -ORs. In another preferred embodiment Ri is selected from the group consisting of heteroaryl and -(C3- C7)heterocycloalkyl, wherein said heteroaryl, -(C3-C7)heterocycloalkyl are optionally substituted by one or more -(Ci-Ce)alkyl. In a more preferred embodiment Ri is selected from the group consisting of pyridine, morpholine and piperazine, optionally substituted by one or more -(Ci- C4)alkyl.

[0091] In one embodiment, R2 and R3 are independently H or -(Ci-Ce)alkyl, optionally substituted by -ORs, or are fused together in a -(C3-C7)heterocycloalkyl.

[0092] In a preferred embodiment Rz and Rj are independently H or -(Ci-Ce)alkyl, wherein said - (Ci-Ce)alkyl is optionally substituted by -ORs. In a more preferred embodiment, Rz and Rj are independently H or -(Ci-C4)alkyl, wherein said -(Ci-C4)alkyl is optionally substituted by -ORs. In a more preferred embodiment Rz and Rj are independently H or methyl. In another preferred embodiment Rz and Rj are fused together in a -(C3-C7)heterocycloalkyl. In a more preferred embodiment Rz and R3 are fused together in a -(C3-C7)heterocycloalkyl selected from oxetane or tetrahydrofurane. In a even more preferred embodiment R2 and R3 are independently H or methyl, optionally substituted by -OH, or are fused together in a tetrahydrofurane.

[0093] In one embodiment, R4 and R5 are independently H or -(Ci-Ce)alkyl. In a preferred embodiment, R4 and R5 are independently H or -(Ci-C4)alkyl. In a more preferred embodiment, R4 and R5 are independently H or methyl. In a more preferred embodiment, R4 and R5 are H.

[0094] In one embodiment, Re and R7 are fused together in a -(C3-C7)heterocycloalkyl. In a preferred embodiment Re and R7 are fused together in a -(C3-Ce)heterocycloalkyl. In a preferred embodiment Re and R7 are fused together in a -(C3-Ce)heterocycloalkyl selected from pyrrolidine, morpholine, piperidine, piperazine, tetrahydrothiazole. In a more preferred embodiment Re and R7 are fused together in a pyrrolidine.

[0095] In one embodiment A is a ring selected from the group consisting of aryl and heteroaryl, wherein any of such aryl, heteroaryl are optionally substituted by one or more groups selected from halogen, -(Ci-Ce)alkyl, -ORs, -NH(CO)-(Ci-Ce)alkyl, -(CO)-NR4Rs. In a preferred embodiment A is optionally substituted aryl or heteroaryl, wherein heteroaryl is selected from thiophene, dihydrobenzofurane or benzofurane. In a more preferred embodiment A is aryl, optionally substituted by one or more groups selected from halogen, -(Ci-C4)alkyl, -ORs, - NH(CO)-(Ci-C6)alkyl, -(CO)-NR4R5.

[0096] In one embodiment, Li is a bond or selected from the group consisting of -(Ci-Ce)alkylene- , -SO2-(Ci-Ce)alkylene- and -(CO)-NReR7-, wherein when Li is -(CO)-NReR7- and Re and R7 are fused together in a -(C3-C7)heterocycloalkyl, the carboxyl group -(CO)-O-R4 of formula (I) is linked to anyone of the C atom of such -(C3-C7)heterocycloalkyl. In a preferred embodiment Li is a bond or selected from the group consisting of -(Ci-C4)alkylene-, -SO2-(Ci-C4)alkylene- and - (CO)-NReR7-, wherein when Li is -(CO)-NReR7- and Re and R7 are fused together in a -(C3- Ce)heterocycloalkyl, the carboxyl group -(CO)-O-R4 of formula (I) is linked to anyone of the C atom of such -(C3-Ce)heterocycloalkyl.

[0097] In a more preferred embodiment Li is a bond or selected from the group consisting of methylene group, -SCh-methylene and 1,2-pyrrolidin-diyl-carbonyl linker.

[0098] All the preferred groups listed above for each of the variable moieties Xi, X2, Ri, Rz, RJ, R4, Rs, Re, R7, Li and A of the compounds of the invention may be combined with each other in embodiments which are included in the scope of the invention.

[0099] In another preferred embodiment, the invention refers to a compound of formula (I) as PHGDH inhibitor, wherein R4 is H, represented by the general formula (la) wherein Xi, X2, Ri, Rz, R3, Rs, Re, R7, Li and A are defined as above.

[0100] In a preferred embodiment, the invention refers to a compound of formula (I) or a compound of formula (la) as PHGDH inhibitor, wherein:

[0101] Ri is H or selected from the group consisting of -(Ci-C4)alkyl, -(Ci-C4)aminoalkyl, -ORs, - SRs, pyridine, morpholine and piperazine, wherein said -(Ci-C4)aminoalkyl or piperazine optionally substituted by one or more -ORs or -(Ci-C4)alkyl;

[0102] R2 and R3 are independently H or methyl, optionally substituted by -OH, or are fused together in a tetrahydrofurane; and pharmaceutically acceptable salts thereof.

[0103] In another preferred embodiment, the invention refers to a compound of formula (I) as PHGDH inhibitor, wherein Li is -(C0)-NReR7-, Re and R7 are fused together in a -(C3- C7)heterocycloalkyl, represented by the general formula (lb) wherein Xi, X2, Ri, Rz, Rs, Rt, Rs and A are defined as above.

[0104] In a preferred embodiment, the invention refers to at least one of the compounds listed in Table 1 below and pharmaceutically acceptable salts thereof.

[0105] Table 1 - List of preferred compounds It is to be understood that all the single deuterates, enantiomers, diastereoisomers and mixtures thereof, in any proportion, or pharmaceutically acceptable salts and solvates of the compounds of formula (I) are encompassed within the scope of the present invention.

[0106] In a more preferred embodiment, the invention refers to a compound of formula (I) as PHGDH inhibitor, wherein Li is -(C0)-NReR7-, Re and R7 are fused together in a pyrrolidine, represented by the general formula (Ic) wherein Xi, X2, Ri, Rz, Rs, Rt, Rs and A are defined as above.

[0107] In a most preferred embodiment, the invention refers to a compound of formula (I) as PHGDH inhibitor, wherein Li is -(C0)-NReR7-, Re and R7 are fused together in a pyrrolidine and R4 is H, represented by the general formula (Id) wherein Xi, X2, Ri, Rz, R3, Rs and A are defined as above.

[0108] In another preferred embodiment, the invention refers to a compound of formula (I) as PHGDH inhibitor, wherein A is an aryl, optionally substituted by one or more groups selected from halogen, -(Ci-Ce)alkyl, -ORs, -NH(CO)-(Ci-Ce)alkyl, -(C0)-NR4Rs; and pharmaceutically acceptable salts thereof.

[0109] In another preferred embodiment, the invention refers to a compound of formula (I) as PHGDH inhibitor, wherein A is an heteroaryl, optionally substituted by one or more groups selected from halogen or -(Ci-Ce)alkyl; and pharmaceutically acceptable salts thereof. All the preferred groups listed above for each of the variable moieties Xi, X2, Ri, Rz, Rs, R4, Rs, Re, R7, Li and A of the compounds of the invention may be combined with each other in embodiments which are included in the scope of the invention.

[0110] In another embodiment, the invention refers to a compound of formula (I) as PHGDH inhibitor. In this respect, it has been found that the compounds of formula (I) of the present invention have an inhibitor drug potency in the low NAD conditions assay, expressed as pICso on PHGDH receptor, equal or higher than 5.5.

[0111] Preferably, the compounds of the present invention have a pICso on PHGDH in low NAD conditions assay between 5.5 and 6.5. More preferably, the compounds of the present invention have a pICso on PHGDH between 6.5 and 7.5. Most preferably, the compounds of the present invention have a pICso on PHGDH higher than 7.5.

[0112] It also been found that the compounds of formula (I) of the present invention have an inhibitor drug potency in the high NAD conditions assay, expressed as pICso on PHGDH receptor, equal or higher than 4.5.

[0113] Preferably, the compounds of the present invention have a pICso on PHGDH in high NAD conditions assay between 4.5 and 5.5. More preferably, the compounds of the present invention have a pICso on PHGDH between 5.5 and 6.5. Most preferably, the compounds of the present invention have a pICso in high NAD conditions assay on PHGDH higher than 6.5.

[0114] The present invention also refers to a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, in admixture with at least one or more pharmaceutically acceptable carrier and / or excipient.

[0115] As used herein, " 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.

[0116] 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.

[0117] In one embodiment, the invention refers to a pharmaceutical composition of compounds of formula (I) in admixture with one or more pharmaceutically acceptable carrier or excipient, for example those described in Remington’s Pharmaceutical Sciences Handbook, XVII Ed., Mack Pub., N.Y., U.S.A.

[0118] In therapeutic use, the compound of formula (I) may be administered by any convenient, suitable, or effective route. Administration of the compounds of the invention and their pharmaceutical compositions may be accomplished according to patient needs, for example, orally, nasally, parenterally (subcutaneously, intravenously, intramuscularly, intrastemally and by infusion) and by inhalation.

[0119] Preferably, the compounds of the present invention are administered orally or by inhalation.

[0120] In a more preferred embodiment, the compounds of the present invention or their pharmaceutical compositions are administered orally.

[0121] In another preferred embodiment, the compounds of the present invention or their pharmaceutical compositions are administered by inhalation.

[0122] In one preferred embodiment, the pharmaceutical composition comprising the compound of formula (I) is a solid oral dosage form such as tablets, gel caps, capsules, caplets, granules, lozenges and bulk powders.

[0123] 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.

[0124] In a further embodiment, the pharmaceutical composition comprising a compound of formula (I) is a liquid oral dosage form such as aqueous and non-aqueous solution, emulsion, suspension, syrup. Such liquid dosage form 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.

[0125] 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.

[0126] Inhalable preparations include inhalable powders, propellant-containing metering aerosols or propellant-free inhalable formulations and may be administered through a suitable inhalation device which may be respectively selected from dry powder inhaler, pressurized metered dosed inhaler, or a nebulizer.

[0127] In another embodiment the invention is also directed to a device comprising a pharmaceutical composition comprising a compound of formula (I) according to the invention, or a pharmaceutically acceptable salt thereof, obtained as described above according to the invention, in form of a single- or multi-dose dry powder inhaler or a metered dose inhaler.

[0128] 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.

[0129] 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.

[0130] 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.

[0131] The compounds of the invention can be administered as the sole active agent or in combination with other pharmaceutical active ingredients.

[0132] 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.

[0133] In a further aspect, the invention refers to the use of the compounds of formula (I) for the preparation of a medicament. In another aspect, the present invention refers to a compound of formula (I) for use as a medicament.

[0134] 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 3 -phosphoglycerate dehydrogenase (PHGDH) receptors mechanism.

[0135] In a further embodiment, the present invention refers to a compound of formula (I) for use in the prevention and / or treatment of diseases, disorders or conditions associated with dysregulation of PHGDH.

[0136] In one aspect, the invention also refers to a method for the prevention and / or treatment of disorders associated with PHGDH receptors mechanisms, said method comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula (I).

[0137] In one embodiment, the present invention refers to a compound of formula (I) useful for the prevention and / or treatment of fibrosis and / or diseases, disorders, or conditions that involve fibrosis.

[0138] The terms "fibrosis" or "fibrotic 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.

[0139] In a preferred embodiment, the present invention is directed to the compounds of formula (I) for use for the prevention and / or treatment of a fibrotic disease.

[0140] In another embodiment, the present invention provides a method for preventing and / or treating fibrotic diseases, the method comprising administering a compound of formula (I).

[0141] In another embodiment, the present invention provides a method for preventing and / or treating fibrotic diseases, the method comprising administering a pharmaceutical composition comprising the compounds of formula (I).

[0142] In another aspect, the present invention is directed to a pharmaceutical composition comprising the compounds of formula (I) and one or more pharmaceutically acceptable carriers and / or excipients, for use for the prevention and / or treatment of fibrotic diseases.

[0143] In a further aspect, the fibrotic diseases mentioned above are selected from pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), hepatic fibrosis, sarcoidosis, familiar pulmonary fibrosis, chronic hypersensitivity pneumonitis (CHP), kidney or renal fibrosis, ocular fibrosis, cardiac fibrosis, arterial fibrosis and systemic sclerosis.

[0144] Preferably, the compounds of formula (I) of the present invention are useful for the treatment and / or prevention of fibrosis such as pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), hepatic fibrosis, sarcoidosis, familiar pulmonary fibrosis, chronic hypersensitivity pneumonitis (CHP), kidney or renal fibrosis, ocular fibrosis, cardiac fibrosis, arterial fibrosis and systemic sclerosis.

[0145] In another embodiment, the invention refers to the use of the compound of formula (I) or its pharmaceutical composition for the preparation of a medicament for the treatment and / or prevention of pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), hepatic fibrosis, sarcoidosis, familiar pulmonary fibrosis, chronic hypersensitivity pneumonitis (CHP), kidney or renal fibrosis, ocular fibrosis, cardiac fibrosis, arterial fibrosis and systemic sclerosis.

[0146] In another embodiment, the invention refers to a compound of formula (I) or a pharmaceutical composition for use in the prevention and / or treatment of fibrotic disease, wherein the fibrotic disease is selected from: pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), hepatic fibrosis, sarcoidosis, familiar pulmonary fibrosis, chronic hypersensitivity pneumonitis (CHP), kidney or renal fibrosis, ocular fibrosis, cardiac fibrosis, arterial fibrosis and systemic sclerosis.

[0147] In another embodiment, the invention refers to a method for the treatment and / or prevention of fibrotic diseases selected from the group consisting of pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), hepatic fibrosis, sarcoidosis, familiar pulmonary fibrosis, chronic hypersensitivity pneumonitis (CHP), kidney or renal fibrosis, ocular fibrosis, cardiac fibrosis, arterial fibrosis and systemic sclerosis, the method comprising administering a compound of formula (I) or a pharmaceutical composition comprising a compound of formula (I) and one or more pharmaceutically acceptable carriers and / or excipients.

[0148] More preferably, the compounds of formula (I) of the present invention are useful for the treatment of idiopathic pulmonary fibrosis (IPF).

[0149] In a more preferred embodiment, the fibrotic disease mentioned above is IPF.

[0150] In another preferred embodiment, the invention refers to a compound of formula (I) or its pharmaceutical composition for use in the prevention and / or treatment of IPF.

[0151] In another preferred embodiment, the invention refers to the use of the compounds of formula (I) or its pharmaceutical composition for the preparation of a medicament for the treatment and / or prevention of IPF.

[0152] In another preferred embodiment, the invention refers to a method for the treatment and / or prevention of IPF, the method comprising administering a compound of formula (I) or a pharmaceutical composition comprising a compound of formula (I) and one or more pharmaceutically acceptable carriers and / or excipients.

[0153] Any suitable route of administration may be employed for providing a mammal, especially a human, with an effective dosage of the compound of formula (I).

[0154] The magnitude of prophylactic or therapeutic dose of the compound of formula (I) will, of course, vary with the nature of the severity of the condition to be treated and with its route of administration, and will generally be determined by clinical trial as required in the pharmaceutical art. It will also vary according to the age, weight and response of the individual patient.

[0155] All preferred groups or embodiments described above for compounds of formula (I) may be combined with each other and apply as well mutatis mutandis.

[0156] The compounds of the present invention can be prepared in a number of ways known to the one skilled in the art of organic synthesis. It will be understood that the functionality / s 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. While the optimal reaction conditions may vary depending on the reactants or solvent used, such conditions can be readily determined by those skilled in the art by routine optimization procedures. Thus, processes described below should not be viewed as limiting the scope of the synthetic methods available for the preparation of the compounds of the invention. In some cases, generally known protective groups (PG) may be employed when needed to mask or protect sensitive or reactive moieties, in accordance with general principles of chemistry (Protective group in organic syntheses, 3rd ed. T. W. Greene, P. G. M. Wuts). The compounds of formula (I), including all the compounds here above listed, can be generally prepared according to the procedures outlined in Schemes shown below, wherein at least one non-limiting synthetic route is provided for the preparation of the exemplified compounds (i.e. the Examples).

[0157] Exemplified preparation processes are given in the following experimental part.

[0158] PREPARATIONS OF INTERMEDIATES AND EXAMPLES

[0159] Chemical Names of the compounds were generated with Structure To Name Enterprise 10.0 Cambridge Software or are common chemical names. All reagents, for which the synthesis is not described in the experimental part, are either commercially available, or are known compounds or may be formed from known compounds by known methods by a person skilled in the art.

[0160] In the procedures that follow, some of the starting materials are identified through an “Intermediate” or “Example” number with indications on step number. This is provided merely for assistance to the skilled chemist.

[0161] When reference is made to the use of a “similar” or “analogous” procedure, as it 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, that will be appreciated by those skilled in the art.

[0162] List of Abbreviations rt: room temperature

[0163] RT : retention time

[0164] DBU: l,8-Diazabicyclo(5.4.0)undec-7-ene

[0165] DIPEA: diisopropylethyl amine

[0166] THF : tetrahydrofuran

[0167] FCC: flash column chromatography

[0168] RP FCC: reversed phase FCC

[0169] DCM: dichloromethane

[0170] DMSO: dimethyl sulfoxide sat aq: saturated aqueous

[0171] TEA: triethylamine

[0172] TFA: Trifluoroacetic acid

[0173] TLC: Thin-layer chromatography

[0174] HATU: l-[Bis(dimethylamino)methylene]-l / Z-l,2,3-triazolo[4,5-Z>]pyridinium 3-oxide hexafluorophosphate on: overnight

[0175] LC-MS= liquid chromatography / mass spectrometry NMR: nuclear magnetic resonance

[0176] PyBOP: benzotriazol- 1-yl oxytri pyrrolidinophosphonium hexafluorophosphate

[0177] 4-CzIPN: l,2,3,5-Tetrakis(carbazol-9-yl)-4,6-dicyanobenzene

[0178] HPLC = high pressure liquid chromatography

[0179] UPLC = Ultra Performance Liquid Chromatography

[0180] AA powder (L-Asp acid; L-Asparagin; L-Glutamic acid; Hydroxy L-Proline; L-Proline)

[0181] P / S: Penicillin-Streptomycin

[0182] General Synthetic Procedures

[0183] The compounds of the present invention can be prepared in a number of ways known to the one skilled in the art of organic synthesis.

[0184] In one embodiment examples of formula (I) can be prepared according to Scheme 1 starting from suitable aryl chlorides of formula III and amines of formula V by nucleophilic aromatic substitution reactions (SxAr). Such reactions can be performed in organic solvents like for instance 1,4-di oxane in the presence of 1-10 eq of an organic base such as DIPEA, at temperatures up to 120 °C and in pressure resistant sealed vessels in cases when reaction temperature exceeds boiling temperature of the solvent. Aryl chlorides III are commercially available in some instances or can be prepared from corresponding intermediates II by functional group interconversion (FGI) operated by procedures well known by those skilled in the art, for example by treatment with POCh in organic solvents such as DMF at temperatures up to 120 °C. Amines VI are commercially available in some instances or can be prepared by cleavage of corresponding Boc-protected intermediates V by means well known to those skilled in the art, such as treatment with 1-10 equivalents of inorganic (e.g. for instance HC1) or organic (e.g. CF3COOH) acids in suitable organic solvents, such as for example THF, 1,4-di oxane or di chloromethane. Intermediates V in turn can be commercially available or can be prepared from aryl halides of general formula IV by means of a transition metal catalyzed cross-coupling reaction through specific synthetic routes described in detail in the experimental part.

[0185] Examples of formula (la) can be prepared according to Scheme 1 starting from corresponding examples of formula (I), by means of procedures well known to those skilled in the art, for example by hydrolysis. Such hydrolysis can be performed in a water miscible organic solvent, like for instance THF or methanol, in the presence of 1-10 equivalents of a base, such as an inorganic base (e.g. NaOH) in aqueous solution, at temperatures up to 60 °C. Scheme 1

[0186] In another embodiment, examples of formula (I) can be prepared from aryl bromides VIII by means of a transition metal catalyzed cross-coupling reaction, such as for example a Pd- catalyzed reaction (e.g. Suzuki cross-coupling), by using proper boron compounds (e.g. boronic acid), Pd catalyst (e.g. Pd(PPhs)4) and inorganic base (e.g. cesium carbonate), according to Scheme 2.

[0187] Aryl bromides VIII can be prepared from amines of formula VI and aryl bromides of formula VII e.g. by means of S\Ar reactions, in analogy to what described in Scheme 1.

[0188] In some instances, where Ri = (Ci-Ce)alkyl, aryl bromides VII can be prepared from corresponding aryl bromides VII where Ri = H, by means of C-H activation reactions, such as for example photochemical reactions employing suitable A-(acyloxy)-phtalimides, such as for example l,3-dioxoisoindolin-2-yl butyrate, in the presence of suitable acids, such as for example (lS)-(+)-10-camphorsulfonic acid, a suitable photocatalysts, such as for example 2, 4,5,6- tetra(carbazol-9-yl)benzene- 1,3 -dicarbonitrile (4-CzIPN), under irradiation of visible light.

[0189] Examples of formula (I) are then converted to examples of formula (la) as above described, e.g. by means of hydrolysis.

[0190] In some instances, cross-coupling step and hydrolysis step can be telescoped in a one-pot, 2- step process leading to examples of formula (la) directly from aryl bromides VIII, as represented in Scheme 2. In such cases, reagents and conditions to perform cross-coupling are applied until full conversion to examples of formula (I), which are not isolated. Reagents and conditions to perform the second step are then directly applied to the reaction mixture, to lead to examples of formula (la).

[0191] Scheme 2

[0192] In another embodiment, examples of formula (I) can be prepared from aryl halide XI, where W is a halogen selected from F, Cl, Br or I, wherein halide is preferably chloride (W = Cl), by means of a transition metal catalyzed cross-coupling reaction, such as for example a Pd-catalyzed reaction (e.g. Stille cross-coupling), by using proper tin compounds and a Pd catalyst (e.g. Pd(PPh3)4), according to Scheme 2a. Examples of formula (I) are then converted to examples of formula (la) as above described, e.g. by means of hydrolysis. Aryl halide XI can be prepared from aryl bromides X by means of a transition metal catalyzed cross-coupling reaction, such as for example a Pd-catalyzed reaction (e.g. Suzuki cross-coupling), by using proper boron compounds (e.g. boronic acid), Pd catalyst (e.g. Pd(PPh?)4) and inorganic base (e.g. cesium carbonate), according to Scheme 2a. Aryl bromides X can be prepared from amines of formula VI and aryl bromides of formula IX e.g. by means of S\Ar reactions, in analogy to what described in Scheme 1. In another embodiment, examples of formula (la) can be prepared from XI performing a one- pot, 2-step process by means of S\Ar reactions, promoted by an organic or inorganic base, such as for example TEA, DIPEA, K2CO3 or others, in a suitable solvent, such as for example THF, DMF, DMA, 1,4-di oxane, DMSO or mixtures thereof, at room temperature to high temperature, followed by hydrolysis, which can be achieved in a water miscible organic solvent, like for instance THF or methanol, in the presence of 1-10 equivalents of a base, such as an inorganic base (e.g. NaOH) in aqueous solution, at temperatures up to 60 °C. This one-pot, 2-step process can also be performed in two separated steps, first by converting Intermediates XI into examples (I), and then by converting examples (I) into examples (la). In another embodiment, examples of formula (la) can be prepared from Intermediates XI performing a one-pot, 2-step process by means of transition metal catalyzed cross-coupling reaction, such as for example a Pd-catalyzed reaction (e.g. Suzuki cross-coupling), by using proper boron compounds (e.g. boronic acid), Pd catalyst (e.g. Pd(PPhs)4) and inorganic base (e.g. cesium carbonate), followed by hydrolysis, which can be achieved in a water miscible organic solvent, like for instance THF or methanol, in the presence of 1-10 equivalents of a base, such as an inorganic base (e.g. NaOH) in aqueous solution, at temperatures up to 60 °C.

[0193] Scheme 2a

[0194] In another embodiment, Intermediate of formula XVII can be prepared from amine VI and aryl halide XVI, where W is a halogen selected from F, Cl, Br or I, wherein halide is preferably chloride (W = Cl) and Z is hydrogen, by nucleophilic aromatic substitution reactions (SxAr). Such reactions can be performed in organic solvents like for instance 1,4-di oxane in the presence of 1- 10 eq of an organic base such as DIPEA, at temperatures up to 120 °C and in pressure resistant sealed vessels in cases when reaction temperature exceeds boiling temperature of the solvent, according to Scheme 2b.

[0195] Intermediate of formula X, wherein halide is preferably chloride (W = Cl) and Z is bromine, can be prepared from Intermediate of formula XVII, where W is is preferably chloride (W = Cl) and Z is hydrogen, by means of bromination reaction, by using proper compounds such as 1- bromopyrrolidine-2, 5-dione in a suitable solvent, such as for example THF at room temperature.

[0196] Intermediates XII can be prepared according to known procedures (WO 2017 156165) and converted into Intermediates XIII first, then into Intermediates XIV as represented in Scheme 3, by applying known conditions.

[0197] In another embodiment, protected amines of general formula V can be prepared starting from commercially available Intermediates IV through the synthetic sequence reported in Scheme 4, involving a first step of a Pd-catalysed substitution of aryl bromide atom by suitable 2- mercaptoacetate esters, in the presence of a suitable Pd source, such as for example Pd(dppf)C12, a suitable ligand, such as for example XPhos and a suitable base, such as for example TEA, DIPEA or others, followed by oxidation of sulfides of general formula XV to corresponding sulfones of general formula V, in the presence of a suitable oxidating agent, such as for example hydrogen peroxide or others, and in a suitable solvent, such as for example AcOH, or others.

[0198] General Experimental details

[0199] Purifications

[0200] Purification by “chromatography”, “flash chromatography” or “flash column chromatography (FCC)” refers to purification using an Interchim PuriFlash Compact 420 or PuriFlash XS420 purification system, or equivalent MPLC using a pre-packed polypropylene column containing stationary phase (cartridge). Where products were purified using a Si cartridge, this refers to an Interchim (or equivalent) pre-packed polypropylene column containing unbounded activated silica with spherical particles with average size of 50 pm (SIHP 50). Fractions containing the desired product (identified by TLC and / or LC-MS analysis) were pooled and concentrated in vacuo.

[0201] NMR Methods

[0202] NMR spectra were obtained on a Bruker Avance III HD 400 MHz (5 mm RT inverse probe head) or Bruker Fourier 300 HD 300 MHz spectrometers using standard Bruker pulse sequences. DMSO-d6, d4-MeOD or CDCh were used as solvents. Chemical shifts are given relative to internal standard tetramethylsilane or solvent residual peak. All experiments were recorded at 298 °K, unless stated differently. 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=broad, nd=not determined.

[0203] In some cases, signals of exchangeable protons (e.g. NH from amide, amine or aniline group, OH from acidic or alcoholic group) are not visible. In a few cases, some signals could be hidden or partly covered by the signal of water or under the DMSO peak or other residual solvents.

[0204] LC-MS Methods

[0205] Method 1

[0206] Dionex UHPLC Ultimate 3000 with DAD detector / Thermo Scientific MSQ Plus; Column: Kinetex ® 2.6 pm XB C18 (4.6x50mm), 110A, mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile; see Table 2. Table 2-Method 1 conditions

[0207] Column temperature: 25 °C; UV detection: from 190 nm to 340 nm; MS conditions: Ionisation Mode: alternate-scan Positive and Negative Electrospray (ES+ / ES-), Scan Range: 100 to 1000 AMU. Method 2

[0208] Dionex UHPLC Ultimate 3000 with DAD detector / Thermo Scientific MSQ Plus; Column: Kinetex ® 2.6 pm XB C18 (4.6x50mm), 110A, mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile; see Table 3.

[0209] Table 3: Method 2 conditions Column temperature: 25 °C; UV detection: from 190 nm to 340 nm; MS conditions:

[0210] Ionisation Mode: alternate-scan Positive and Negative Electrospray (ES+ZES-), Scan Range: 100 to 1000 AMU.

[0211] Method 3

[0212] Dionex UHPLC Ultimate 3000 with DAD detector / Thermo Scientific MSQ Plus; Column: Kinetex ® 2.6 pm XB C18 (4.6x50mm), 110A, mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile; see Table 4. Table 4: Method 3 conditions

[0213] Column temperature: 25 °C; UV detection: from 190 nm to 350 nm; MS conditions: Ionisation Mode: alternate-scan Positive and Negative Electrospray (ES+ / ES-), Scan Range: 100 to 1000 AMU. Method 4

[0214] Dionex UHPLC Ultimate 3000 with DAD detector / Thermo Scientific MSQ Plus; Column: Kinetex ® 2.6 pm XB C18 (4.6x50mm), 110A, mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile; see Table 5.

[0215] Table 5: Method 4 conditions Column temperature: 25 °C; UV detection: from 190 nm to 350 nm; MS conditions:

[0216] Ionisation Mode: alternate-scan Positive and Negative Electrospray (ES+ZES-), Scan Range: 100 to 1000 AMU.

[0217] Method 5

[0218] Shimadzu LCMS-2020 Single Quadrupole Liquid Chromatograph Mass Spectrometer; Column: Acquity UPLC BEH - Waters, 1.7 pm C18 (2.1 x 100 mm), 130 A), mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile; see Table 6. Table 6: Method 5 conditions

[0219] Column temperature: 25 °C; UV detection: 220 nm and 254 nm; MS conditions: Ionisation Mode: alternate-scan Positive and Negative Electrospray (ES+ZES-), Scan Range: 100 to 1000 AMU. Method 6

[0220] Shimadzu LCMS-2020 Single Quadrupole Liquid Chromatograph Mass Spectrometer; Column: Acquity UPLC BEH - Waters, 1.7 pm C18 (2.1 x 100 mm), 130 A), mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile; see Table 7.

[0221] Table 7: Method 6 conditions Column temperature: 25 °C; UV detection: 220 nm and 254 nm; MS conditions: Ionisation

[0222] Mode: alternate-scan Positive and Negative Electrospray (ES+ / ES-), Scan Range: 100 to 1000 AMU.

[0223] Method 7

[0224] Waters ACQUITY UPLC I-Class PLUS System with Waters SQ Detector 2; Column: Acquity UPLC BEH C18 1.7 pm (2.1 x 100 mm), mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile; see Table 8. Table 8: Method 7 conditions

[0225] Column temperature: 40 °C; UV detection: from 200 nm to 400 nm; MS conditions: Ionisation Mode: alternate-scan Positive and Negative Electrospray (ES+ / ES-), Scan Range: 100 to 1000 AMU. Method 8

[0226] Waters ACQUITY UPLC I-Class PLUS System with Waters SQ Detector 2; Column: Acquity UPLC BEH C18 1.7 pm (2.1 x 100 mm), mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile; see Table 9.

[0227] Table 9: Method 8 conditions Column temperature: 40 °C; UV detection: from 200 nm to 400 nm; MS conditions:

[0228] Ionisation Mode: alternate-scan Positive and Negative Unispray, Scan Range: 100 to 1000 AMU.

[0229] Method 9

[0230] Dionex UHPLC Ultimate 3000 with DAD detector / Thermo Scientific MSQ Plus; Column: Kinetex ® 2.6 pm XB C18 (4.6x50mm), 110A, mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile; see Table 10. Table 10: Method 9 conditions

[0231] Column temperature: 25 °C; UV detection: from 190 nm to 350 nm; MS conditions: Ionisation Mode: alternate-scan Positive and Negative Electrospray (ES+ / ES-), Scan Range: 100 to 1000 AMU. Method 10

[0232] Dionex UHPLC Ultimate 3000 with DAD detector / Thermo Scientific MSQ Plus; Column: Kinetex ® 2.6 pm XB C18 (4.6x50mm), 110A, mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile; see Table 11.

[0233] Table 11: Method 10 conditions Column temperature: 25 °C; UV detection: from 190 nm to 350 nm; MS conditions:

[0234] Ionisation Mode: alternate-scan Positive and Negative Electrospray (ES+ZES-), Scan Range: 100 to 1000 AMU.

[0235] Method 11

[0236] Dionex UHPLC Ultimate 3000 with DAD detector / Thermo Scientific MSQ Plus; Column: Kinetex ® 2.6 pm XB C18 (4.6x50mm), 110A, mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile; see Table 12. Table 12: Method 11 conditions

[0237] Column temperature: 25 °C; UV detection: from 190 nm to 340 nm; MS conditions: Ionisation Mode: alternate-scan Positive and Negative Electrospray (ES+ZES-), Scan Range: 100 to 1000 AMU. Method 12

[0238] Dionex UHPLC Ultimate 3000 with DAD detector / Thermo Scientific MSQ Plus; Column: Kinetex ® 2.6 pm XB C18 (4.6x50mm), 110A, mobile phase A: 0.1% formic acid in water, mobile phase B: 0.1% formic acid in acetonitrile; see Table 13.

[0239] Table 13: Method 12 conditions Column temperature: 25 °C; UV detection: from 190 nm to 350 nm; MS conditions:

[0240] Ionisation Mode: alternate-scan Positive and Negative Electrospray (ES+ / ES-), Scan Range: 100 to 1000 AMU.

[0241] Method 13

[0242] Acquity UPLC coupled with SQD mass spectrometer; Column: Acquity BEH C18 (100mm x 2.1mm i.d., 1.7pm), mobile phase A: 0.1% (v / v) formic acid in water, mobile phase B: 0.1% (v / v) formic acid in acetonitrile; see Table 14. Table 14: Method 13 conditions

[0243] Column temperature: 40 °C; UV detection: from 210 nm to 400 nm; MS conditions: Ionisation Mode: alternate- scan Positive and Negative Electrospray (ES+ZES-), Scan Range: 100 to 1500 AMU.

[0244] Method 14

[0245] Kinetex®2.6 pm XB-C18 (4.6x50mm), 110A, column temperature was 25 °C, mobile phase A was 0.1 % v / v water solution of formic acid, mobile phase B 0.1 % v / v acetonitrile solution of formic acid. The flow rate was 1 mL / min. The UV detection range was 190-350 nm ± 4 nm and ES+ZES- range was 100 to 1000

[0246] AMU. See Table 15.

[0247] Table 15: Method 14 conditions

[0248] Method 15

[0249] Acquity CSH C18 column 50mm x 2.1mm 1.7pm, maintained at 40°C; Mobile Phase: Eluent B (ACN) in Eluent A (water +0.1% HCOOH) from 1% to 99.9% within 1.5 min. Flow rate: 1 mL / min. Wavelength: 210-400 nm DAD. UPLC + Waters PDA + Waters QDA.

[0250] Method 16 Acquity UPLC coupled with SQD mass spectrometer; Column: Acquity BEH Cl 8 (50mm x 2.1mm i.d., 1.7pm), mobile phase A: 0.1% (v / v) formic acid in water, mobile phase B: 0.1% (v / v) formic acid in acetonitrile; see Table 16.

[0251] Table 16: Method 16 conditions

[0252] Column temperature: 40 °C; UV detection: from 210 nm to 400 nm; MS conditions: Ionisation Mode: alternate-scan Positive and Negative Electrospray (ES+ZES-), Scan Range: 100 to 1500 AMU.

[0253] PREPARATION OF INTERMEDIATES:

[0254] Intermediates V

[0255] Intermediate V-l: ethyl (l?)-2-((4-(l-aminoethyl)phenyl)sulfonyl)acetate hydrochloride

[0256] Step 1: tert-butyl (l?)-(l-(4-bromophenyl)ethyl)carbamate

[0257] A solution of (7?)-l-(4-bromophenyl)ethan-l-amine (3.45 mL, 24 mmol), BOC2O (6.38 g, 29 mmol) and EtsN (6.70 mL, 48 mmol) in DCM (50 mL) was stirred overnight at rt. The mixture was diluted with DCM (150 mL) and washed with brine (2 X). Organic layer was dried over Na2SO4, filtered and concentrated under reduced pressure. Purification by FCC (gradient elution from 100:0 to 50:50 hexane / EtOAc) yielded title compound (4.69 g, 14 mmol, 59% yield).

[0258] LC-MS Method 5: RT 4.064 min ( 254 nm), MS ESI (+) m / z = 326.2 [M+Na]+.

[0259] Step 2: ethyl (l?)-2-((4-(l-((tert-butoxycarbonyl)amino)ethyl)phenyl)thio)acetate

[0260] A reaction tube was charged with tert-butyl ( / )-(! -(4-bromophenyl)ethyl)carbam ate (2 g, 6.66 mmol), Pd(dppf)C12 (487 mg, 0.67 mmol) and Xantphos (578 mg, 0.999 mmol), sealed with septum and purged with Ar (4 X vacuum-fill cycles). 1,4-dioxane (3 mL), DIPEA (2.32 mL, 13.32 mmol) and ethyl 2-mercaptoacetate (2.19 mL 19.99 mmol) were then added. The mixture was stirred on at 120 °C. Volatiles were evaporated, then the residue was suspended in water (10 mL) and extracted with EtOAc (3 X 15 mL). Combined organic extracts were washed with brine (20 mL), dried over Na2SO4 and concentrated under reduced pressure. Purification by FCC (isocratic elution with EtOAc / hexane 15:85) yielded title compound (1.45 g, 3.63 mmol, 55% yield).

[0261] LC-MS Method 5: RT 3.36 min (X 254 nm), MS ESI (+) m / z = 239.5 [M-Boc]+.

[0262] Step 3: ethyl (l?)-2-((4-(l-((tert-butoxycarbonyl)amino)ethyl)phenyl)sulfonyl)acetate

[0263] H2O2 (0.70 mL, 8.97 mmol) was slowly added to a stirred solution of ethyl (A)-2-((4-(l- ((tert-butoxycarbonyl)amino)ethyl)phenyl)thio)acetate (870 mg, 2.56 mmol) in AcOH (5 mL). Stirring went on at 60 °C. The reaction mixture was allowed to cool to rt, then volatiles were removed under reduced pressure. The residue was dissolved in EtOAc and washed with H2O. Concentration under reduced pressure followed. FCC (eluting with DCM / MeOH 95:5) yielded title compound as a white solid (440 mg, 0.569 mmol, 22% yield).

[0264] LC-MS Method 7: RT 1.60 min, MS (ESI) m / z = 370.48 [M-H]’.

[0265] Intermediate V-5: methyl (4-((l?)-l-((tert-butoxycarbonyl)amino)ethyl)benzoyl)-£- prolinate

[0266] (A)-4-(l-((tert-butoxycarbonyl)amino)ethyl)benzoic acid (6.93 g, 26.1 mmol) and methyl L- prolinate hydrochloride (5.19 g, 31.3 mmol) were dissolved in DCM (130 mL). TEA (14.56 mL, 104 mmol) was added followed by HATU (14.90 g, 39.2 mmol). The reaction mixture was stirred overnight at rt. The mixture was diluted with DCM and sat aq NaHCCh. The resulting mixture was stirred for 15 min, then layers were separated. Water layer was extracted twice with DCM. Combined organic layers were washed with water, then brine, dried over Na2SO4, filtered and concentrated under reduced pressure. Purification by FCC (DCM / MeOH 100:0 to 90: 10 gradient elution) yielded title compound (10.9 g, 89% purity, contamination by 11% w / w tetramethylurea, 25.8 mmol, 99% yield).

[0267] LC-MS Method 7: RT 1.57 min, MS (ESI) m / z = 377.4 [M+H]+. Intermediates VI

[0268] Intermediate VI- 1: ethyl (l?)-2-((4-(l-aminoethyl)phenyl)sulfonyl)acetate hydrochloride

[0269] 4 M HC1 in 1,4 dioxane (0.28 mL, 1.131 mmol) was added to a solution of (7?)-2-((4-(l- ((tert-butoxycarbonyl)amino)ethyl)phenyl)sulfonyl)acetate (420 mg, 1.131 mmol) in anhydrous DCM (10 mL). Stirring went on overnight at rt. Precipitate was filtered off, washed with DCM and dried under vacuum to yield title compound (282 mg, 0.926 mmol, 82% yield).

[0270] LC-MS Method 8: RT 1.24 min, MS (ESI) m / z = 255.42 [M-NH2]+.

[0271] Intermediate VI-5: methyl (4-((l?)-l-aminoethyl)benzoyl)-£-prolinate

[0272] 4N HC1 in 1,4-di oxane (36 mL, 144 mmol) was added to a solution of methyl (4-(( / ?)- l - ((tert-butoxycarbonyl)amino)ethyl)benzoyl)-Z-prolinate (10.85 g, 28.8 mmol) in DCM (60 mL). The reaction mixture was stirred at rt overnight. Dilution with DCM and sat aq NaHCCh followed and this mixture was stirred for 10 min. The layers were separated and water layer was extracted twice with DCM. Combined organic layers were washed with water and brine, then dried over Na2SO4, filtered and concentrated under reduced pressure. Purification by FCC (100:0 to 90: 10 DCM / MeOH gradient elution) yielded title compound as a yellow oil (5.35 g, 19.4 mmol, 67% yield).

[0273] LC-MS Method 7: RT 1.24 min, MS (ESI) m / z = 277.2 [M+H]+.

[0274] Intermediate VI-33 : methyl 4-(3-aminotetrahydrofuran-3-yl)benzoate

[0275] Step 1: methyl 4-(3-cyanotetrahydrofuran-3-yl)benzoate

[0276] To a solution of NaH (0.685 g, 17.12 mmol) in DMF (15 mL) was added methyl 4- (cyanomethyl)benzoate (1 g, 5.7 mmol) and l-chloro-2-(chloromethoxy)ethane (0.59 mL, 5.99 mmol) in DMF (15 mL) dropwise at -20 °C under argon atmosphere. The reaction mixture was stirred at rt on. The mixture was poured into water and extracted with ethyl acetate. The organic layers were washed with brine. Solvents were removed in vacuo and crude material was purified by FCC (hexane:ethyl acetate, 98:2 to 95:5) to give desired product (300 mg, 1.3 mmol, 23 % yield).

[0277] 'H NMR (300 MHz, DMSO-d6) 5 8.07 - 7.99 (m, 2H), 7.73 - 7.65 (m, 2H), 4.41 (dd, J = 9.0, 0.7 Hz, 1H), 4.14 - 4.05 (m, 2H), 3.91 (d, J = 8.9 Hz, 1H), 3.87 (s, 3H), 2.86 - 2.76 (m, 1H), 2.59 - 2.54 (m, 1H).

[0278] Step 2: methyl 4-(3-carbamoyltetrahydrofuran-3-yl)benzoate

[0279] To a solution of methyl 4-(3-cyanotetrahydrofuran-3-yl)benzoate (300 mg, 1.3 mmol) in DMSO (5 mL) were added potassium carbonate (71.7 mg, 0.52 mmol) and hydrogen peroxide (1.06 mL, 10.4 mmol). The reaction mixture was stirred at rt for 3 hr. Sodium sulfite was added to the RM and extracted with ethyl acetate. The organic layers were washed with water and concentrated.

[0280] Crude material was triturated with hexane to give title compound (280 mg, 1.12 mmol, 87 % yield).XH NMR (300 MHz, DMSO-d6) 5 7.97 - 7.89 (m, 2H), 7.51 - 7.43 (m, 2H), 7.35 (s, 1H), 7.12 (s, 1H), 4.49 (d, J = 8.6 Hz, 1H), 3.85 (s, 3H), 3.84 - 3.75 (m, 3H), 2.91 - 2.81 (m, 1H), 2.20 - 2.09 (m, 1H).

[0281] Step 3: methyl 4-(3-aminotetrahydrofuran-3-yl)benzoate

[0282] To a mixture of methyl 4-(3 -carbarn oyltetrahydrofuran-3-yl)benzoate (280 mg, 1.12 mmol) in Acetonitrile (5 mL) and Water (1.0 mL) was added Bis(trifluoroacetoxy)iodobenzene (580 mg, 1.35 mmol). The reaction mixture was stirred at rt on. Water and DCM were added, then aqueous layer was acidify and extracted with DCM. The organic layer was passed through phase separator. Solvents were removed in vacuo.

[0283] Crude material was used directly to the next step without further purification (153 mg, 0,69 mmol, 62 % yield).XH NMR (300 MHz, DMSO-d6) 5 7.96 - 7.88 (m, 2H), 7.70 - 7.62 (m, 2H), 4.10 - 3.98 (m, 1H), 3.94 (td, J = 8.3, 3.8 Hz, 1H), 3.85 (s, 3H), 3.79 - 3.69 (m, 2H), 2.27 - 2.15 (m, 1H), 2.11 (s, 2H), 2.09 - 2.01 (m, 1H).

[0284] Intermediates VII

[0285] Intermediate VII-39: 6-bromo-4-chloro-2-propylthieno [2, 3-d] pyrimidine

[0286] A solution of 6-bromo-4-chlorothieno[2,3-d]pyrimidine (300 mg, 1.20 mmol), (1 S)-(+)-10- camphorsulfonic acid (559 mg, 2.40 mmol), l,3-dioxoisoindolin-2-yl butyrate (421 mg, 1.8 mmol) and 4-CzIPN (95 mg, 0.12 mmol) in DMA (5 mL) was added, the mixture bubbled vigorously with nitrogen, sealed, and placed between two blue Kessil LED Photoredox light PR160L lamps (390-456 nm, 352mW / cm2). The mixture was stirred at 50°C overnight. The mixture was partitioned between ethyl acetate and water, the organic phase dried over Na2SO4 and filtrated. The solution was concentrated under reduced pressure and the resulting crude purified by FCC (Heptane: ethyl acetate from 100:0 to 0: 100). Proper fractions were collected to give title compound (25 mg, 0.09 mmol, 7 % yield).

[0287] LC-MS Method 15: RT 1.48 min ( 254 nm), MS (ESI) m / z = 293.01 [M+H]+

[0288] Intermediates VIII

[0289] Intermediates VIII in the following Table 17 were prepared from suitable reagents in analogy to the procedure followed for Intermediate 1-1.

[0290] Table 17: Intermediates VIII

[0291] Intermediate VIII-39: methyl (R)-4-(l-((6-bromo-2-propylthieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoate

[0292] DIPEA (0.075 ml, 0.43 mmol) was added to a mixture of Intermediate VII-39 (25 mg, 0.09 mmol)), PyBOP (45 mg, 0.09 mmol), DBU (0.258 ml, 1.72 mmol) and methyl (R)-4-(l- aminoethyl)benzoate hydrochloride (185 mg, 0.857 mmol) in dioxane (2 mL). The reaction mixture was stirred at 50 °C on.

[0293] The mixture was concentrated under reduced pressure and the residue partitioned between DCM and water. The organic phase was filtered through an hydrophobic phase separator and concentrated under reduced pressure. The crude was purified by FCC (heptane: ethyl acetate from 100:0 to 0: 100) to give title compound (20 mg, 0.05 mmol, 54 % yield).

[0294] LC-MS Method 15: RT 1.39 min ( 254 nm), MS (ESI) m / z = 436.01 [M+H]+

[0295] Intermediates X and XVII

[0296] The Intermediates in the following Table 18 were prepared from suitable reagents in analogy to the procedure followed for Intermediate I- 1. Table 18: Intermediates X and XVII

[0297] Intermediate X-45: ethyl 2-[4-[3-[(6-bromo-2-chloro-thieno[2,3-d]pyrimidin-4- yl)amino]tetrahydrofuran-3-yl]phenyl]-3-methyl-butanoate

[0298] Intermediate XVII-45 ethyl 2-[4-[3-[(2-chlorothieno[2,3-d]pyrimidin-4- yl)amino]tetrahydrofuran-3-yl]phenyl]-3-methyl-butanoate (140 mg, 0.30 mmol) was dissolved in THF (6 mL) and 1 -bromopyrrolidine-2, 5-dione (81.3 mg, 0.46 mmol) was added. Reaction mixture was stirred at rt overnight. Mixture was diluted with EtOAc and water. Organic layer was dried and concentrated to get crude, which was purified by FCC (EtOAc in cyclohexane 0-50%) to get desired product (125 mg, 0,23 mmol, 76 % yield). LC-MS Method 16: RT 1.52 min (X 254 nm), MS (ESI) m / z = 538 / 540 [M+H]+

[0299] The Intermediates in the following Table 19 were prepared from suitable reagents in analogy to the procedure followed for Intermediate 1-8.

[0300] Table 19: Intermediates XI Intermediate XIII-38: ethyl 2-(4-(3-carbamoyltetrahydrofuran-3-yl)phenyl)acetate

[0301] This intermediate was prepared following known procedures (WO 2017 156165) to give title compound (279 mg, 1.0 mmol, 65 % yield).

[0302] 'H NMR (300 MHz, DMSO-d6) 5 7.27 (d, J = 8.6 Hz, 2H), 7.22 (d, J = 8.4 Hz, 2H), 4.49 (d, J = 8.5 Hz, 1H), 4.14 - 4.01 (m, 2H), 4.05 - 3.94 (m, 1H), 3.84 - 3.75 (m, 1H), 3.70 (d, J = 8.3 Hz, 1H), 3.63 (s, 2H), 2.81 (d, J = 6.7 Hz, 1H), 2.09 (s, 1H), 1.18 (t, J = 7.1 Hz, 3H).

[0303] Intermediate XIV-38: ethyl 2-(4-(3-aminotetrahydrofuran-3-yl)phenyl)acetate

[0304] Starting from XIII-38, this intermediate was prepared following known procedures (WO 2017 156165) to give title compound (143 mg, 0.57 mmol, 58 % yield).

[0305] Crude material was purified by FCC (gradient from 50 to 100 AcOEt in Hex, then eluent was shifted to DCM:MeOH (1 :4)).

[0306] 'HNMR (300 MHz, DMSO-d6) 5 7.44 (d, J = 8.3 Hz, 2H), 7.31 (d, J = 8.3 Hz, 2H), 4.12 - 4.05 (m, 3H), 4.03 (d, J = 6.2 Hz, 1H), 3.95 (td, J = 8.8, 4.8 Hz, 1H), 3.78 (d, J = 9.3 Hz, 1H), 3.68 (s, 2H), 2.43 - 2.29 (m, 2H), 1.19 (t, J = 7.1 Hz, 3H).

[0307] Intermediates I

[0308] Intermediate 1-1: ethyl (l?)-2-((4-(l-((6-phenylthieno[3,2-J|pyrimidin-4- yl)amino)ethyl)phenyl)sulfonyl)acetate

[0309] DIPEA (0.17 mL, 0.973 mmol) was added to a stirred solution of 4-chloro-6- phenylthieno[3,2-t ]pyrimidine (80 mg, 0.324 mmol, commercially available) and intermediate VI-1 (97 mg, 0.315 mmol) in 1,4-dioxane (5 mL). Stirring went on overnight at 100 °C. Volatiles were evaporated under reduced pressure. FCC (DCMZEtOAc 100:0 to 90: 10 gradient elution) yielded title compound (95 mg, 0.181 mmol, 56% yield).

[0310] LC-MS Method 8: RT 1.51 min, MS (ESI) m / z = 482.48 [M+H]+. 1H NMR (300 MHz, DMSO-d6): 5 8.42 (d, J = 7.5 Hz, 1H), 8.36 (s, 1H), 7.88 (s, 4H), 7.83 (s, 1H), 7.70 (d, J = 8.4 Hz, 2H), 7.59 - 7.44 (m, 3H), 5.66 - 5.51 (m, 1H), 3.97 (q, J = 7.1 Hz, 2H), 1.58 (d, J = 7.1 Hz, 3H), 0.94 (t, J = 7.1 Hz, 3H).

[0311] The Intermediates in the following Table 20 were prepared from suitable reagents in analogy to the procedure followed for Intermediate I- 1.

[0312] Table 20: Intermediates I

[0313] Intermediate 1-8: methyl (4-((7?)-l-((6-(m-tolyl)thieno[3,2-J|pyrimidin-4- yl)amino)ethyl)benzoyl)-Z-prolinate

[0314] The mixture of Intermediate VIII-8 (150 mg, 0.307 mmol,), m-tolylboronic acid (62.5 mg, 0.460 mmol) and cesium carbonate (200 mg, 0.613 mmol) in THF (3 mL) and water (1 mL) was purged with Ar for 15 minutes. Subsequently, Pd(PPhs)4 (35.4 mg, 0.031 mmol) was added. The resulting solution was stirred 10 h at 100 °C. The reaction mixture was then extracted with DCM and washed with brine. Organic layer was dried over anhydrous Na2SO4 filtered and concentrated. Purification by FCC (DCM / MeOH 95:5 isocratic elution) yielded title compound (117 mg, 0.234 mmol, 76% yield).

[0315] LC-MS Method 8: RT 1.54 min (X 254 nm), MS ESI(+) m / z = 501.8 [M+H]+.

[0316] 1H NMR (300 MHz, DMSO-d6) 5 8.38 (s, 1H), 8.32 (d, J = 7.9 Hz, 1H), 7.78 (s, 1H), 7.69 (s, 1H), 7.64 (d, J = 7.9 Hz, 1H), 7.50 (s, 4H), 7.46 - 7.20 (m, 3H), 5.62 - 5.43 (m, 1H), 4.45 (dd, J = 8.2, 4.5 Hz, 1H), 3.65 (s, 3H), 3.51 (t, J = 6.1 Hz, 2H), 2.40 (s, 3H), 2.26 (d, J = 6.2 Hz, 1H), 1.92 - 1.79 (m, 4H), 1.57 (d, J = 7.0 Hz, 3H).

[0317] The Intermediates in the following Table 21 were prepared from suitable reagents in analogy to the procedure followed for Intermediate 1-8. Table 21: Intermediates I

[0318] The Intermediates in the following Table 22 were prepared from suitable reagents in analogy to the procedure followed for Intermediate V-5.

[0319] Table 22 Intermediate 1-32: methyl (4-((R)-l-((6-(2-fluoro-6-methoxyphenyl)-2-(pyridin-2- yl)thieno[2,3-d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L-prolinate

[0320] To a mixture of Intermediate XI-20 methyl (4-((R)-l-((2-chloro-6-(2-fluoro-6- methoxyphenyl)thieno[2,3-d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L-prolinate (50 mg, 0.09 mmol) and Tetrakis(triphenylphosphine)palladium(0) (10.15 mg, 8.8 pmol) under argon, a solution of 2-(tributylstannyl)pyridine (28.1 pl, 0.09 mmol) in anhydrous dioxane (0.5 mL) was added. The mixture was stirred under argon on in 120 °C. Reaction mixture was poured on sat. aq. KF solution and stirred Ih, then mixture was extracted with EtOAc. Organic layer was evaporated and extracted with ACN / Pentane. ACN fraction was evaporated and crude was purified by FCC (DCM to DCM: NFE-sat-MeOH 95:5) to give desired product (43 mg, 0.07 mmol, 80% yield).

[0321] LC-MS Method 7: RT 1.45 min ( 254 nm), MS (ESI) m / z = 612.7 [M+H]+

[0322] 'H NMR (300 MHz, DMSO-d6) 5 8.69 (d, J = 4.7 Hz, IH), 8.55 (d, J = 7.5 Hz, IH), 8.24 (d, J = 7.9 Hz, IH), 8.17 (s, IH), 7.90 (td, J = 7.5, 1.4 Hz, IH), 7.56 (dd, 4H), 7.49 - 7.42 (m, 2H), 7.12 - 6.99 (m, 2H), 5.72 (t, J = 7.1 Hz, IH), 4.49 - 4.35 (m, IH), 3.94 (s, 3H), 3.64 (s, 3H), 3.48

[0323] (t, J = 6.4 Hz, 2H), 2.25 - 2.16 (m, IH), 1.90 - 1.75 (m, 3H), 1.64 (d, J = 7.1 Hz, 3H).

[0324] The Intermediate in the following Table 23 was prepared from suitable reagents in analogy to the above procedure.

[0325] Table 23 Intermediate 1-35: methyl (4-((R)-l-((6-(2-fluoro-6-methoxyphenyl)-2-

[0326] (methylamino)thieno[2,3-d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L-prolinate

[0327] Step 1: methyl (4-((R)-l-((6-(2-fluoro-6-methoxyphenyl)-2-((4- methoxybenzyl)(methyl)amino)thieno[2,3-d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L- prolinate

[0328] The solution of Intermediate XI-20 methyl (4-((R)-l-((2-chloro-6-(2-fluoro-6- methoxyphenyl)thieno[2,3-d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L-prolinate (60 mg, 0.11 mmol) and l-(4-methoxyphenyl)-N-methylmethanamine (112 pL, 0.74 mmol) in Dioxane (3 mL) was stirred on at 120 °C.

[0329] The reaction mixture was extracted with EtOAc and washed with brine. Organic layer was dried over anhydrous Na2SO4, filtered and concentrated. The crude product was purified by FCC (Hex to Hexane:Ethyl Acetate 3:7) to give title compound (63 mg, 0.09 mol, 87 % yield).

[0330] LC-MS Method 7: RT 2.0 min ( 254 nm),MS (ESI) m / z = 684.2 / 685.2 [M+H]+

[0331] 'HNMR (300 MHz, DMSO-d6) 5 8.10 (d, J = 7.3 Hz, 1H), 7.88 (d, J = 1.4 Hz, 1H), 7.50 - 7.18 (m, 5H), 7.09 - 6.90 (m, 4H), 6.77 (d, J = 8.4 Hz, 2H), 5.38 (t, J = 7.0 Hz, 1H), 4.85 (d, J = 15.2 Hz, 1H), 4.45 (dd, J = 8.1, 4.6 Hz, 2H), 3.88 (s, 3H), 3.69 (s, 3H), 3.65 (s, 3H), 3.44 (dt, J = 10.5, 4.0 Hz, 2H), 2.90 (d, J = 8.7 Hz, 3H), 2.21 (s, 1H), 1.83 (d, J = 6.1 Hz, 3H), 1.53 (d, J = 7.1 Hz, 3H).

[0332] Step 2: methyl (4-((R)-l-((6-(2-fluoro-6-methoxyphenyl)-2-(methylamino)thieno[2,3- d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L-prolinate

[0333] To a solution of methyl (4-((R)-l-((6-(2-fluoro-6-methoxyphenyl)-2-((4- methoxybenzyl)(methyl)amino)thieno[2,3-d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L-prolinate (43 mg, 0.06 mmol) in DCM (3 mL), TFA (0.145 mL, 1.89 mmol) was added and stirred at 50 °C for 2 days. Reaction mixture was extracted with sat. aq. NaHCCh. Organic layer was dried over Na2SO4 and evaporated to give desired product (35mg, quantitative yield). LC-MS Method: RT 1.68 min (X 254 nm), MS (ESI) m / z =565.0 [M+H]+

[0334] The Intermediates I in the following Table 24 were prepared from suitable reagents in analogy to the procedure used applied to the synthesis of Intermediate VIII-39.

[0335] Table 24: Intermediates I

[0336] PREPARATION OF EXAMPLES:

[0337] Example (l?)-2-((4-(l-((6-phenylthieno[3,2-J|pyrimidin-4- yl)amino)ethyl)phenyl)sulfonyl)acetic acid 1 MNaOH (0.59 mL, 0.592 mmol) was added to a solution of intermediate 1-1, 95 mg, 0.197 mmol) in THF (5 mL). Stirring went on overnight at rt. Reaction mixture was concentrated under reduced pressure. The residue was redissolved in water, followed by washing with DCM. The aqueous layer was acidified to pH 6.5. The precipitate was washed with water (3 X), then dried in vacuo to yield title compound (8 mg, 0.017 mmol, 9% yield). LC-MS Method 3: RT 2.39 min, MS (ESI) m / z = 453.86 [M+H]+.

[0338] 1H NMR (300 MHz, DMSO-d6): 5 8.39 (d, J = 8.2 Hz, 2H), 7.91 - 7.84 (m, 2H), 7.84 - 7.78 (m, 3H), 7.61 - 7.43 (m, 5H), 5.57 (t, J = 7.2 Hz, 1H), 3.75 (s, 2H), 1.58 (d, J = 7.1 Hz, 3H).

[0339] In the following Table 25 the Examples were prepared from suitable reagents in analogy to the procedure followed for Example 1. Table 25

[0340]

[0341] Example 9: (4-((l?)-l-((6-(2,3-dihydrobenzofuran-7-yl)thieno[3,2-J|pyrimidin-4- yl)amino)ethyl)benzoyl)-Z-proline

[0342] A mixture of Intermediate VIII-8 (100 mg, 0.20 mmol), (2,3-dihydrobenzofuran-7- yl)boronic acid (50.3 mg, 0.31 mmol) and cesium carbonate (133 mg, 0.409 mmol) in THF (1533 pl) and water (511 pl) was purged with Ar for 15 minutes. Subsequently, tetrakis(triphenylphosphine)palladium(0) (11.81 mg, 10.22 pmol) was added. The resulting solution was stirred at 90 °C on. The reaction mixture was allowed to cool down to rt, then IM NaOH (2043 pl, 2.043 mmol) was added and reaction mixture was stirred at rt overnight. Volatiles were evaporated under reduced pressure, then water was added to the residue. 1 M HC1 was added until pH —2 / 3 was reached, and a precipitate formed. After trituration in water and drying overnight in Genevac equipment at 40 °C, title compound was obtained (48 mg, 0.093 mmol, 46% yield).

[0343] LC-MS Method 4: RT 3.23 min ( 254 nm), MS (ESI) m / z = 356.88 [M-H]

[0344] ‘H NMR (300 MHz, DMSO-d6) 5 12.86 (s, 1H), 9.10 (d, J = 7.8 Hz, 1H), 7.97 - 7.87 (m, 2H), 7.58 - 7.49 (m, 3H), 7.36 (d, J = 0.8 Hz, 1H), 7.30 - 7.23 (m, 1H), 7.19 (dd, J = 7.5, 0.9 Hz, 1H), 5.21 (p, J = 7.2 Hz, 1H), 3.97 (s, 3H), 1.51 (d, J = 7.1 Hz, 3H). The Examples in the following Table 26 were obtained from suitable reagents in analogy to the procedure followed for Example 9. Details of conditions are provided in table when significantly different from above.

[0345] Table 26

[0346]

[0347] Example 29: (4-((l?)-l-((6-(3-fluoro-5-methylphenyl)-2-morpholinothieno[2,3- |pyrimidin-4-yl)amino)ethyl)benzoyl)-Z-proline

[0348] To a solution of Intermediate XI-28 methyl (4-((R)-l-((2-chloro-6-(3-fluoro-5- methylphenyl)thieno[2,3-d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L-prolinate (60 mg, 0.11 mmol) and morpholine (0.066 mL, 0.76 mmol) in Dioxane (3 mL), DIPEA (0.038 mL, 0.217 mmol) was added and mixture was stirred on at 100 °C. To the reaction mixture 8.0 eq of 8M NaOH was added and stirred on in 60 °C. Organic solvent was evaporated and basic water solution acidified by IM HC1 with precipitation of a solid which was collected and was purified by prep-HPLC to give title compound (41 mg, 0.07 mmol, 64 % yield).

[0349] LC-MS Method 12: RT 3.82 min ( 254 nm), MS (ESI) m / z = 590.28 [M+H]+

[0350] 'H NMR (300 MHz, DMSO-d6) 5 12.55 (s, 1H), 8.07 (d, J = 7.2 Hz, 1H), 8.03 (s, 1H), 7.48 (s, 4H), 7.24 (s, 1H), 7.15 (d, J = 10.2 Hz, 1H), 6.98 (d, J = 9.6 Hz, 1H), 5.34 (t, J = 7.0 Hz, 1H), 4.37

[0351] (dd, J = 8.1, 4.5 Hz, 1H), 3.69 - 3.45 (m, 10H), 2.37 (s, 3H), 2.25 - 2.12 (m, 1H), 1.96 - 1.72 (m, 3H), 1.55 (d, J = 6.9 Hz, 3H).

[0352] The Example 31 in the following Table 27 was prepared from suitable reagents in analogy to the procedure followed for Example 29.

[0353] Table 27

[0354] Example 20: (4-((l?)-l-((6-(2-fluoro-6-methoxyphenyl)-2-(methylthio)thieno[2,3- |pyrimidin-4-yl)amino)ethyl)benzoyl)-£-proline

[0355] To the solution of Intermediate XI-20 methyl (4-((R)-l-((2-chloro-6-(2-fluoro-6- methoxyphenyl)thieno[2,3-d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L-prolinate (50 mg, 0.09 mmol) in 1,4-Dioxane (5 mL), sodium thiomethoxide (18.5 mg, 0.26 mmol) was added and mixture was stirred at 70 °C overnight. Reaction mixture was acidified by addition of IM HC1 and extracted with DCM. Organic layer was dried over Na2SO4 and evaporated, to give 50 mg of solid. The crude was purified by prep-HPLC to give title compound (11 mg, 0.02 mmol, 22 % yield). LC-MS Method 1 : RT 3.39 min (X 254 nm), MS (ESI) m / z = 566.9 [M+H]+

[0356] 1HNMR (300 MHz, DMSO-d6) 5 12.57 (s, 1H), 8.50 (d, J = 7.8 Hz, 1H), 8.05 (s, 1H), 7.55 - 7.33 (m, 5H), 7.05 (d, J = 8.0 Hz, 1H), 6.99 (d, J = 9.5 Hz, 1H), 5.49 (t, J = 7.1 Hz, 1H), 4.36 (d, J = 5.2 Hz, 1H), 3.91 (s, 3H), 3.48 (d, J = 6.6 Hz, 2H), 2.39 (s, 3H), 2.27 (s, 1H), 1.93 - 1.74 (m, 3H), 1.56 (d, J = 6.4 Hz, 3H).

[0357] Example 23: (4-((l?)-l-((6-(2-fluoro-6-methoxyphenyl)-2-methoxythieno[2,3- |pyrimidin-4-yl)amino)ethyl)benzoyl)-Z-proline

[0358] Intermediate XI-20 methyl (4-((R)-l-((2-chloro-6-(2-fluoro-6-methoxyphenyl)thieno[2,3- d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L-prolinate (100 mg, 0.18 mmol) was dissolved in dry MEOH (2 mL) and sodium methanolate (xx ml, 0.5 mmol) was added under argon and reaction mixture was stirred at 70 °C on. Then IM NaOH (2 ml, 2.0 mmol) was added and reaction mixture was stirred at rt overnight. Volatiles were evaporated under reduced pressure, then water was added to the residue. 1 M HC1 was added until pH —2 / 3 was reached, and a precipitate formed. Filtration yielded title compound (14 mg, 0.025 mmol, 14% yield).

[0359] LC-MS Method 4: RT 3.14 min ( 254 nm), MS (ESI) m / z = 550.86 [M+H]+

[0360] ‘H NMR (300 MHz, DMSO-d6) 5 12.51 (s, 1H), 8.44 (d, J = 7.7 Hz, 1H), 8.01 (s, 1H), 7.54 - 7.30 (m, 5H), 7.03 (d, J = 8.1 Hz, 1H), 6.98 (d, 1H), 5.47 (t, J = 7.3 Hz, 1H), 4.37 (dd, J = 8.2, 4.8 Hz, 1H), 3.90 (s, 3H), 3.78 (s, 3H), 3.60 - 3.40 (m, 2H), 2.25 - 2.15 (m, 1H), 1.98 - 1.71 (m, 3H), 1.57 (d, J = 6.9 Hz, 3H).

[0361] The Example 28 in the following Table 28 was prepared from suitable reagents in analogy to the procedure followed for Example 23.

[0362] Table 28

[0363] PHARMACOLOGICAL ACTIVITY OF THE COMPOUNDS OF THE INVENTION 3-Phosphoglycerate Dehydrogenase (PHGDH) Biochemical Assay

[0364] This enzymatic assay measures inhibitory activity of compounds of PHGDH enzyme, which catalyzes the reaction of 3 -phosphoglycerate and NAD+to 3-phosphohydroxypyruvate and NADH. The NADH, produced by the reaction, is used in a coupled reaction of reduction resazurin to resorufin by diaphorase; the resorufin is measured in a fluorescence intensity readout. The assay was performed using commercially available recombinant Human PHGDH, full length, C- terminal His-tag (BPS Biosciences 71079). The assay was adapted for two different NAD+final assay concentrations: low NAD: 4 pM, high NAD: 250 pM. Two mixes were prepared before running the protocol: Mix 1 : with PHGDH (final assay concentration 0.008 pg / ml) and NAD+(final assay concentration 4 pM or 250 pM) in the assay buffer (Tris 41.7 mM HC1, pH 7.5, 18.7 mM hydrazine sulfate pH 9, 0.8 mM EDTA, 0.004% Tween20) and Mix 2: with 0.17 mM PGA, 0.02 mM resazurin and 0.66 U / mL diaphorase in H2O.

[0365] The enzymatic reaction was performed by incubating each concentration of compounds with MIX 1 at 25 °C. After an incubation of 60 min, the MIX 2 was added and further incubated at 25°C for 180 min. After the incubation, the fluorescence intensity was read (Excitation @ X 554 nm / Emission @ X 593 nm). The assays were performed in 384-well format and validated using a selected reference compound, tested in 10-point concentration-response curve. All compounds were tested in 10-point concentration-response curve, 3 -fold dilutions, each in duplicate. The following controls were included in each assay plate: high control - DMSO vehicle control (1% final reaction volume) and low control - maximal inhibition control (3 pM reference inhibitor). Raw data were processed to obtain dose response curves for the compounds: Fluorescence data are normalized to high control and low control and IC50 values were determined in automated script for dose-response analysis, using 4 parameter model: log(inhibitor) vs. response. Results were expressed as pICso value (negative logarithm of ICso). QC criteria parameters: Z' > 0.5, Hill Slope range 0.5 to 5, S:B > 2.

[0366] Test compound potencies in low NAD are reported in Table 29 as pIC50 values.

[0367] Table 29: pICso values in Low NAD+concentration assay wherein the compounds are classified in term of potency with respect to their inhibitory activity on PHGDH enzyme according to the following classification criterion:

[0368] +: pICso comprised between 5.5 and 6.5

[0369] ++: pICso comprised between 6.5 and 7.5

[0370] +++: pICso higher than 7.5

[0371] As it can be appreciated, all the compounds of Table 29 show an inhibitory activity on PHGDH enzyme. In fact, it can be recognized that the symbol + indicate a sufficient or good level of activity, which can be even increased up to +++, thus confirming the high activity on PHGDH enzyme of the compounds of the invention.

[0372] Compounds according to the invention showed PHGDH inhibitory activity higher than 5.5 pICso (lowNAD+conditions assay), corresponding to < 3 pM in terms of inhibitory concentration. Most preferably, compounds showed values higher than 6.5, and even more preferably higher than 7.5, corresponding to < 0.3 pM, even more preferably < 30 nM, in terms of inhibitory concentration.

[0373] 13C3- Serine flux Assay in MDA-MB-468 Cells

[0374] This method describes a cellular serine flux assay based on13C3-Serine MS read out to profile PHGDH inhibitors. PHGDH is a key enzyme for Serine biosynthesis, and it is described that breast cancer cells (MDA-MB468) can be dependent on PHGDH for their growth and survival. Two different media, without glycine and serine, were used for the Serine flux assay in the cell plates:

[0375] 1) starting medium: modified RPMI 1640 (Teknova R9660) supplemented with AA powder, P / S, 5 mM / .-Glutamine, 10% FBS dialyzed, 2 g / L D-(+)-glucose. 2) assay medium: modified RPMI 1640 supplemented with AA powder, P / S, 5 mM L- Glutamine, 10% dialyzed FBS, 2 g / L13Ce-glucose, instead of unlabelled Z>-(+)-Glucose.

[0376] On assay day 0, MDA-MB-468 cells were seeded at a density of 50.000 cells / well in 100 pL starving medium lacking Serine and Glycine and incubated for 20-24 hours. Day 1, cells were washed twice with assay medium containing13Ce-Glucose before addition of compound dilutions; compound solutions were prepared by a further dilution in DMSO from stock to 30 mM, Serial dilution 1 :3 in DMSO was performed in a compound-plate, to generate 8 points Concentration- Response-Curve (CRC), each point in triplicate.

[0377] Compound-plates were used immediately after preparation on the day of the experiment. The following controls were included in assay plate: basal control / no inhibition (DMSO, 8- replicates / plate); QC / maximal inhibition (reference compound, 30 pM final, 8-replicates / plate). Assay cell plates were then incubated at 37 °C, 5% CO2 for 24 hours, when the conditioned culture medium was used for13C3-Serine detection. On Day 2, 50 pL / well of the cell supernatant was collected and transferred into a sample-plate for LC-MS analysis. Raw data were processed to obtain dose response curves for the compounds: data were normalized to high control and low control and IC50 values were determined in automated script for dose-response analysis, using 4 parameter model: log(inhibitor) vs. response.

[0378] Results were expressed as IC50 value, given in nM for each NCE or pICso (negative logarithm of IC50). QC criteria parameters: Z' > 0.5, Hill Slope range 0.5 to 5, S:B > 2.

Claims

CLAIMS1. A compound of formula (I):wherein at least one of Xi and X2 is S, and the other one is CH;Ri is H or selected from the group consisting of -(Ci-Ce)alkyl, -(Ci-C6)aminoalkyl, - ORs, -SRs, heteroaryl and -(C3-C7)heterocycloalkyl, wherein said -(Ci-C6)aminoalkyl, heteroaryl, -(C3-C7)heterocycloalkyl are optionally substituted by one or more group selected from -(Ci-Ce)alkyl or -ORs;R2 and Rj are independently H or -(Ci-Ce)alkyl, optionally substituted by -ORs, or are fused together in a -(C3-C7)heterocycloalkyl;R4 and R5 are independently H or -(Ci-Ce)alkyl;A is a ring selected from the group consisting of aryl and heteroaryl, wherein any of such aryl, heteroaryl are optionally substituted by one or more groups selected from halogen, -(Ci-Ce)alkyl, -ORs, -NH(CO)-(Ci-C6)alkyl, -(CO)-NR4R5;LI is a bond or selected from the group consisting of -(Ci-Ce)alkylene-, -SO2-(Ci- Ce)alkylene- and -(C0)-NReR7-, wherein when Li is -(C0)-NReR7- and Re and R7 are fused together in a -(C3- C7)heterocycloalkyl, the carboxyl group -(C0)-0-R4 of formula (I) is linked to anyone of the C atom of such -(C3-C7)heterocycloalkyl; and pharmaceutically acceptable salts thereof.

2. The compound of formula (I) according to claim 1, wherein R4 is H, represented by the general formula (la)whereinXi, X2, Ri R2, Rs, Rs, Re, R7, A and Li are as defined in claim 1; and pharmaceutically acceptable salts thereof.

3. The compound of formula (I) according to claim 1 or 2, wherein:Ri is H or selected from the group consisting of -(Ci-C4)alkyl, -(Ci-C4)aminoalkyl, - ORs, -SRs, pyridine, morpholine and piperazine, wherein said -(Ci-C4)aminoalkyl or piperazine optionally substituted by one or more -ORs or -(Ci-C4)alkyl;Rz and R3 are independently H or methyl, optionally substituted by -OH, or are fused together in a tetrahydrofurane; and pharmaceutically acceptable salts thereof.

4. The compound according to any one of claims 1 to 3, selected from at least one of(R)-2-((4-(l-((6-phenylthieno[3,2-d]pyrimidin-4- yl)amino)ethyl)phenyl)sulfonyl)acetic acid;(R)-2-(4-(l-((6-phenylthieno[3,2-d]pyrimidin-4-yl)amino)ethyl)phenyl)acetic acid;(R)-4-(l-((6-phenyl-2-(pyri din-2 -yl)thieno[2, 3-d]pyrimidin-4- yl)amino)ethyl)benzoic acid;(R)-4-(l-((6-phenyl-2-(pyridin-3-yl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoic acid;(4-((R)-l-((6-phenyl-2-(pyridin-2-yl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(2-methoxyphenyl)thieno[2,3-d]pyrimidin-4-yl)amino)ethyl)benzoyl)- L-proline;(4-((R)-l-((6-phenyl-2-(pyridin-3-yl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(m-tolyl)thieno[3,2-d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(2,3-dihydrobenzofuran-7-yl)thieno[3,2-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(3-chloro-5-methylphenyl)thieno[3,2-d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(2,3-difluorophenyl)thieno[3,2-d]pyrimidin-4-yl)amino)ethyl)benzoyl)- L-proline;(4-((R)-l-((6-(2-fluoro-6-methoxyphenyl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(benzofuran-7-yl)thieno[3,2-d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L- proline;(4-((R)-l-((6-(5-fluoro-2-methoxyphenyl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(2,3-dihydrobenzofuran-7-yl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((2-((2-methoxyethyl)amino)-6-(2-methoxyphenyl)thieno[2,3-d]pyrimidin- 4-yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(2-methoxyphenyl)-2-(pyridin-3-yl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(2-methoxyphenyl)-2-morpholinothieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(S)-4-(2-hydroxy-l-((6-phenylthieno[3,2-d]pyrimidin-4-yl)amino)ethyl)benzoic acid;(4-((R)-l-((6-(2-fluoro-6-methoxyphenyl)-2-(methylthio)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(2-fluoro-6-methoxyphenyl)-2-(pyridin-3-yl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(3-fluoro-5-methylphenyl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(2-fluoro-6-methoxyphenyl)-2-methoxythieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(2-methoxyphenyl)-2-(4-methylpiperazin-l-yl)thieno[2,3-d]pyrimidin- 4-yl)amino)ethyl)benzoyl)-L-proline;(R)-4-(l-((2-methyl-6-phenylthieno[2,3-d]pyrimidin-4-yl)amino)ethyl)benzoic acid;(4-((R)-l-((6-(3-fluoro-5-methylphenyl)thieno[3,2-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((2-methyl-6-phenylthieno[2,3-d]pyrimidin-4-yl)amino)ethyl)benzoyl)-L- proline;(4-((R)-l-((6-(3-fluoro-5-methylphenyl)-2-methoxythieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(3-fluoro-5-methylphenyl)-2-morpholinothieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(5-chlorothi ophen-2 -yl)thieno[2, 3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(2-fluoro-6-methoxyphenyl)-2-morpholinothieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(2-fluoro-6-methoxyphenyl)-2-(pyri din-2 -yl)thieno[2, 3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;4-(3-((6-phenylthieno[3,2-d]pyrimidin-4-yl)amino)tetrahydrofuran-3-yl)benzoic acid;(4-((R)-l-((6-(3-fluoro-5-methylphenyl)-2-(pyri din-2 -yl)thieno[2, 3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(2-fluoro-6-methoxyphenyl)-2-(methylamino)thieno[2,3-d]pyrimidin- 4-yl)amino)ethyl)benzoyl)-L-proline;(R)-4-(l-((6-(2-fluoro-6-methoxyphenyl)-2-(pyridin-3-yl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoic acid;(R)-4-(l-((6-(3-fluoro-5-methylphenyl)-2-(pyridin-3-yl)thieno[2,3-d]pyrimi din-4- yl)amino)ethyl)benzoic acid;2-(4-(3-((6-(2-fluoro-6-methoxyphenyl)-2-(pyridin-3-yl)thieno[2,3-d]pyrimidin-4- yl)amino)tetrahydrofuran-3-yl)phenyl)acetic acid;(R)-4-(l-((6-phenyl-2-propylthieno[2,3-d]pyrimidin-4-yl)amino)ethyl)benzoic acid;(4-((R)-l-((6-(2-(methylcarbamoyl)phenyl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(2-acetamidophenyl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(3-acetamidophenyl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;(4-((R)-l-((6-(3-(methylcarbamoyl)phenyl)thieno[2,3-d]pyrimidin-4- yl)amino)ethyl)benzoyl)-L-proline;3-methyl-2-(4-(3-((6-phenylthieno[3,2-d]pyrimidin-4-yl)amino)tetrahydrofuran-3- yl)phenyl)butanoic acid;2-(4-(3-((6-(2-fluoro-6-methoxyphenyl)-2-(pyridin-3-yl)thieno[2,3-d]pyrimidin-4- yl)amino)tetrahydrofuran-3-yl)phenyl)-3 -methylbutanoic acid; as single deuterate, enantiomer, diastereoisomer or mixtures thereof, in any proportion, or pharmaceutically acceptable salts and solvates thereof.

5. The compound of formula (I) according to claim 1-3, wherein Li is -(C0)-NReR7-, Re and R? are fused together in a pyrrolidine, represented by the general formula (Ic)wherein Xi, X2, Ri, Rz, Rs, Rt, Rs and A are as defined in claim 1-3; and pharmaceutically acceptable salts thereof.

6. The compound of formula (I) according to claim 5, wherein R4 is hydrogen.

7. The compound of formula (I) according to claim 1-6, wherein:A is an aryl, optionally substituted by one or more groups selected from halogen, -(Ci- C6)alkyl, -OR5, -NH(CO)-(Ci-C6)alkyl, -(CO)-NR4R5; and pharmaceutically acceptable salts thereof.

8. The compound of formula (I) according to claim 1-6, wherein:A is an heteroaryl, optionally substituted by one or more groups selected from halogen or -(Ci-Ce)alkyl; and pharmaceutically acceptable salts thereof.

9. A pharmaceutical composition comprising a compound according to any one of claims 1 to 8, in admixture with one or more pharmaceutically acceptable carrier or excipient.

10. A compound of formula (I) according to any one of claims 1-8 or a pharmaceutical composition according to claims 9 for use as a medicament.

11. A compound of formula (I) according to any one of claims 1-8 or a pharmaceutical composition according to claims 9 for use according to claim 10 in treating disease, disorder, or condition associated with dysregulation of 3 -phosphoglycerate dehydrogenase (PHGDH).

12. A compound of formula (I) according to claims 1 to 8 or a pharmaceutical composition according to claims 9 for use according to claim 10 in the prevention and / or treatment of fibrosis and / or diseases, disorders, or conditions that involve fibrosis.

13. A compound of formula (I) or a pharmaceutical composition for use according to claim 12 in the prevention and / or treatment of fibrosis including pulmonary fibrosis, idiopathic pulmonary fibrosis (IPF), hepatic fibrosis, sarcoidosis, familiar pulmonary fibrosis, chronichypersensitivity pneumonitis (CHP), kidney or renal fibrosis, ocular fibrosis, cardiac fibrosis, arterial fibrosis and systemic sclerosis.

14. A compound of formula (I) or a pharmaceutical composition for use according to claim 13 in the prevention and / or treatment idiopathic pulmonary fibrosis (IPF).

15. The pharmaceutical composition for use according to claim 10-14 for oral administration.

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