HK40067957A - Thienopyrimidine derivatives as lpa receptor 2 inhibitors - Google Patents

Description

Thienopyrimidine derivatives as LPA receptor 2 inhibitors

Technical Field

The present invention relates generally to compounds that inhibit lysophosphatidic acid receptors (hereinafter LPA inhibitors); the present invention relates to compounds which are thienopyrimidine derivatives, to processes for preparing such compounds, to pharmaceutical compositions containing them and to their therapeutic use.

The compounds of the present invention may be used, for example, in the treatment of a number of disorders associated with the mechanisms of the LPA receptor.

Background

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

These LPA-mediated processes are involved in nervous system function, vascular development, immune system function, cancer, reproduction, fibrosis, and obesity (see, e.g., Yung et al, J Lipid Res.2014 7 months; 55(7): 1192-. LPA species are formed depending on their precursor phospholipid, which can generally vary with acyl chain length and saturation. The term LPA generally denotes 18:1 oleoyl-LPA (1-acyl-2-hydroxy-sn-glycerol 3-phosphate), which is the most abundant form of LPA in human plasma, with 16:0-, 18:2-, and 18:1-LPA (see, e.g., Sano et al, J Biol chem.2002Dec 13; 277(50): 21197-. All LPA species are produced from membrane phospholipids through two major metabolic pathways. Depending on the site of synthesis, membrane phospholipids are converted to the corresponding lysophospholipids by the action of phospholipase a1(PLA1), phospholipase a2(PLA2) or PLA1 and lecithin-cholesterol acyltransferase (LCAT). Autotaxin (ATX) then acts on lysophospholipids and converts them to LPA species. The second pathway first converts phospholipids to phosphatidic acid by the action of phospholipase D. PLA1 or PLA2 then metabolized the phosphatidic acid to lysophosphatidic acid (see, e.g., Riaz et al, Int J Mol Sci.2016, 2 months; 17(2): 215).

ATX activity is the major source of plasma extracellular LPA, but the source of tissue LPA contributing to the signaling sink may not only involve ATX but also other enzymes. The biological functions of LPA are mediated by at least six recognized cell surface receptors.

All LPA receptors are rhodopsin-like 7-TM proteins that signal through at least two of the four G α subunit families (G α 12/13, G α q/11, G α i/o and G α S). LPA receptors often trigger the response of a variety of heterotrimeric G proteins, producing different results in an environment and cell type dependent manner. G α 12/13-mediated LPA signaling regulates cell migration, invasion and cytoskeletal remodelling through the activation of RHO pathway proteins. RAC activation downstream of G α i/o-PI3K also regulates a similar process, but the most prominent functions of LPA-induced G α i/o are mitogenic signaling through the RAF-MEK-MAPK cascade and survival signaling through the PI3K-AKT pathway. LPA-coupled G α q/11 proteins regulate Ca2+ homeostasis primarily via PLC and the second messengers IP3 and DAG. Finally, G.alpha.S can activate adenylate cyclase and increase cAMP concentration following LPA stimulation (see, e.g., Riaz et al, Int J Mol Sci.2016, 2 months; 17(2): 215).

LPAs, especially LPA1, LPA2 and LPA3, have been involved in migration, invasion, metastasis, proliferation and survival and differ in their tissue distribution and downstream signaling pathways.

LPA1 is a 41-kD protein that is widely expressed in all examined human adult tissues, albeit at different levels, and the importance of LPA1 signaling in development and adult life has been demonstrated by numerous protocols (see, e.g., Ye et al, 2002, neuroreport.12.3.d.; 13(17): 2169-75). Extensive expression of LPA1 was observed in adult mice, with significant presence in at least brain, uterus, testis, lung, small intestine, heart, stomach, kidney, spleen, thymus, placenta, and skeletal muscle. LPA1 is also widely expressed in humans, and expression is more spatially restricted during embryonic development. LPA1 couples and activates three types of G proteins: g alpha i/o, G alpha q/11 and G alpha 12/13. LPA1 activation induces a range of cellular responses: cell proliferation and survival, cell migration, cytoskeletal changes, Ca2+ mobilization, adenylate cyclase inhibition, and activation of mitogen-activated protein kinases, phospholipase C, Akt, and Rho pathways (see, e.g., Choi et al, Annu Rev Pharmacol toxicol.2010; 50: 157-86).

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

LPA2 mainly activates the same pathway triggered by LPA1 in terms of signaling activity, with some exceptions, i.e., its unique cross-talk (cross-talk) behavior. For example, LPA2 promotes cell migration through interaction with the focal adhesion molecule TRIP6 (see, e.g., Lai YJ,2005, mol.cell.biol.25:5859-68), and several PDZ proteins and zinc finger proteins have also been reported to interact directly with the carboxy-terminal tail of LPA2 (see, e.g., Lin FT,2008, biochim.biophysis.acta 1781: 558-62).

Human LPA3 is a 40-kD protein and has sequence homology to LPA1 (about 54%) and LPA2 (about 49%). In adults LPA3 is highly expressed in the heart, pancreas, prostate and testes. Moderate levels of expression are also found in the brain, lung and ovary. Like LPA1 and LPA2, the signaling activity of LPA3 stems from its coupling to G α i/o and G α q/11 (see, e.g., Ishii et al, Mol Pharmacol 58:895-902, 2000). Each LPA has a number of important regulatory functions throughout the body.

Since LPA signaling has been closely associated with many disease states, there has been great interest in developing specific LPA inhibitors (see, e.g., Stoddard et al, Biomol Ther (Seoul)2015, month 1; 23(1): 1-11). Various studies have demonstrated a positive role for LPA in the pathogenesis of Pulmonary Fibrosis (PF), a destructive disease characterized by damage to alveolar epithelial cells, accumulation of myofibroblasts, and deposition of extracellular matrix proteins, leading to loss of lung function and death (see, e.g., Wilson MS, Wynn TA (2009), muccosal Immunol 2: 103-.

Evidence suggests that there is a significant increase in lysophosphatidic acid levels in bronchoalveolar lavage fluid of patients with PF, where it acts to mediate fibroblast migration in damaged lungs via LPA1 (see, e.g., Tager et al, Nat med.2008, 1 month; 14(1): 45-54). Furthermore, mice lacking LPA1 or LPA2 were significantly protected from fibrosis and death in a mouse model of bleomycin-induced pulmonary fibrosis (see, e.g., Huang et al, Am J Respir Cell Mol biol.2013, 12 months; 49(6): 912-.

In vitro, LPA1 is known to induce proliferation and differentiation of lung fibroblasts (see, e.g., Shiomi et al, Wund Repair Regen.2011, 3-4 months; 19(2):229-240), and to enhance the contraction of fibroblast-mediated released collagen gels (see, e.g., Mio et al, Journal of Laboratory and Clinical Medicine, Vol.139, No. 1, 2002, 1 month, pages 20-27). In human lung fibroblasts, knockdown of LPA2 attenuates LPA-induced TGF- β 1 expression and differentiation of lung fibroblasts into myofibroblasts, resulting in decreased expression of different profibrotic markers such as FN, α -SMA and collagen, as well as decreased activation of the extracellular regulated kinases 1/2, Akt, Smad3 and p38 mitogen-activated protein kinase (see, e.g., Huang et al, Am J Respir Cell Mol biol.2013, 12 months; 49(6): 912-) -922). Furthermore, Xu et al demonstrated that expression of LPA2 is also upregulated in the lungs of bleomycin-challenged mice, where it is able to induce activation of the TGF- β pathway through the RhoA and Rho kinase pathways, TGF- β being a key cytokine playing an important role in the development of the disease (see, e.g., Xu et al, Am J pathol, 2009, 4 months; 174(4): 1264-79). Oral administration of LPA1 antagonist significantly reduced bleomycin-induced pulmonary fibrosis in mice in an in vivo preclinical model (Tager et al, Nat med.2008, month 1; 14(1): 45-54; Swaney et al, Br J pharmacol.2010, month 8; 160(7):1699-1713), and intraperitoneal injection of LPA1/3 antagonist improved radiation-induced pulmonary fibrosis (see, e.g., Gan et al, 2011, Biochem biophysis Res commu 409: 7-13). In a renal fibrosis model, LPA1 administration of an LPA1 antagonist inhibits renal interstitial fibrosis (see, e.g., Pradere et al, J Am Soc Nephrol 2007; 18: 3110-3118).

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

WO2019126086 and WO2019126087(Bristol-Myers Squibb) disclose cyclohexanoic acid iso-isomers as LPA1 antagonistsAzole-oxazines for use in the treatment of disorders or conditions associated with dysregulation of lysophosphatidic acid receptor 1.

WO2019126099(Bristol-Myers Squibb) discloses isotypes as LPA1 antagonistsAn oxazole N-linked carbamoyl cyclohexanoic acid for use in the treatment of a disorder or condition associated with dysregulation of lysophosphatidic acid receptor 1.

WO2019126090(Bristol-Myers Squibb) discloses triazole N-linked carbamoyl cyclohexanoic acids as LPA1 antagonists. The compounds are selective LPA1 receptor inhibitors and are useful for treating disorders or conditions associated with dysregulation of lysophosphatidic acid receptor 1.

WO2017223016(Bristol-Myers Squibb) discloses carbamoyloxymethyltriazole cyclohexanoic acid as LPA1 antagonist for the treatment of fibrosis, including idiopathic pulmonary fibrosis.

WO2012028243(Merck) discloses pyrazolopyridinone derivatives according to formula (I) as LPA2 receptor antagonists and processes for their preparation for the treatment of various diseases.

Amgen inc. at "Discovery of effective LPA2(EDG4) antagonists as potential anti-cancer agents (Discovery of patent LPA2(EDG4) as potential anti-cancer agents)" Bioorg Med Chem lett.2008, 2/1; LPA2 antagonists are disclosed in (18), (3) 1037-41. Key compounds were evaluated in vitro for inhibition of LPA 2-mediated Erk activation and HCT-116 cell proliferation. These compounds can be used as tool compounds to evaluate the anti-cancer effect of blocking LPA2 signaling.

It is noteworthy that antagonizing LPA receptors may be useful in the treatment of fibrosis and diseases, disorders, and conditions caused by fibrosis, and that even more antagonizing LPA2 may be particularly effective in the treatment of the above-mentioned diseases, disorders, and conditions.

Over the past few years, several efforts have been made to develop novel LPA1 receptor antagonists that can be used to treat a variety of diseases, and some of those compounds have also shown efficacy in humans.

Thus, there remains the potential to develop inhibitors of the receptor LPA2 that may be useful in the treatment of diseases or disorders associated with dysregulation of the LPA receptor, in particular fibrosis.

In this respect, the prior art does not describe or suggest that the thienopyrimidine derivatives of general formula (I) of the present invention have antagonistic activity on the receptor LPA2, which represents a solution to the above-mentioned need.

Disclosure of Invention

In a first aspect, the invention relates to compounds of formula (I)

Wherein

R is H or selected from (C)₁-C₄) Alkyl, (C)₁-C₄) Haloalkyl and 5-6 membered heteroaryl, wherein each of said heteroaryl may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, halo (halo) and (C)₁-C₄) A haloalkyl group;

R₁is H or (C)₁-C₄) An alkyl group;

R₂is H or is selected from (C)₁-C₄) Alkyl, halo, (C)₁-C₄) Haloalkyl and (C)₃-C₈) A cycloalkyl group;

R₃is H or (C)₁-C₄) An alkyl group;

a is selected from 5-6 membered heteroaryl and arylWherein each of said heteroaryl and aryl groups may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, -C (O) R₁、-C(O)OR₁、-C(O)R₁、(C₁-C₄) Haloalkyl, halo, -NR_AC(O)R₁、-NR_AC(O)OR₁、-NR_AC(O)-(C₁-C₄) alkylene-OR₁、-NR_AC(O)R_C、-NR_AC(O)NR_AR_B、-N(C_1-C₄) alkylene-NR_AR_BAryl and heteroaryl, optionally substituted by one or more (C)₁-C₄) Alkyl and (C)₁-C₄) Haloalkyl is substituted, or

When a is aryl, it may be fused to a second saturated or unsaturated ring optionally containing one or more heteroatoms selected from N, O and S to form a bicyclic ring system, optionally substituted with one or more heteroatoms selected from-c (o) R₁、(C₁-C₄) Alkyl and oxo;

R_Cselected from heteroaryl, aryl, (C)₃-C₈) Cycloalkyl and (C)₄-C₈) Heterocycloalkyl, wherein said heteroaryl, aryl, heterocycloalkyl and cycloalkyl may optionally be substituted by one or more (C)₁-C₄) Alkyl and-C (O) OR₁Substitution;

R_Aand R_BIndependently at each occurrence is H or is selected from (C)₁-C₄) Alkyl, (C)₃-C₈) Cycloalkyl, (C)₁-C₆) Haloalkyl and halo, or

R_AAnd R_BMay form, together with the nitrogen atom to which they are attached, a 4-6 membered saturated heterocyclic ring system optionally containing another heteroatom selected from N, S or O, which may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, (C)₁-C₄) A haloalkyl group and a halo group,

provided that when R is methyl and A is aryl, the aryl is not substituted with one or more methyl groups and chlorine.

In a second aspect, the present invention relates to a pharmaceutical composition comprising a compound of formula (I) in admixture with one or more pharmaceutically acceptable carriers or excipients.

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

In another aspect, the present invention relates to compounds of formula (I) for use in the treatment of a disease, disorder or condition associated with dysregulation of lysophosphatidic acid receptor 2(LPA 2).

In another aspect, the present invention relates to compounds of formula (I) for use in the prevention and/or treatment of fibrosis and/or a disease, disorder or condition involving fibrosis.

In another aspect, the present invention relates to compounds of formula (I) for use in the prevention and/or treatment of Idiopathic Pulmonary Fibrosis (IPF).

Detailed Description

Unless otherwise indicated, the compounds of formula (I) of the present invention are also intended to include stereoisomers, tautomers, or pharmaceutically acceptable salts or solvates thereof.

The term "pharmaceutically acceptable salt" as used herein denotes derivatives of the compounds of formula (I) wherein the parent compound is suitably modified as follows: any free acidic or basic groups that may be present are converted to the corresponding addition salts with any base or acid conventionally considered pharmaceutically acceptable.

Suitable examples of such salts may thus include inorganic or organic acid addition salts of basic residues such as amino groups and inorganic or organic base addition salts of acidic residues such as carboxyl groups.

Cations of inorganic bases that may suitably be used to prepare the salts include ions of alkali or alkaline earth metals such as potassium, sodium, calcium or magnesium.

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

The term "solvate" refers to a physical association of a compound of the invention with one or more solvent molecules (whether organic or inorganic). The physical association includes hydrogen bonding. In some cases, the solvate will be capable of isolation, for example, when one or more solvent molecules are incorporated into the crystal lattice of the crystalline solid. The solvate may comprise a stoichiometric or non-stoichiometric amount of solvent molecules.

The term "stereoisomers" refers to isomers that are structurally identical but differ in the arrangement of their atoms in space. Enantiomers and diastereomers are examples of stereoisomers.

The term "enantiomer" means one of a pair of molecular species that are mirror images of each other and that do not overlap.

The term "diastereomer" refers to a stereoisomer that is not a mirror image.

The term "racemate" or "racemic mixture" means a composition consisting of equimolar amounts of two enantiomeric species, wherein the composition is not optically active.

The symbols "R" and "S" represent the configuration of the substituent around the chiral carbon atom. The isomer descriptors "R" and "S" are used herein to indicate the atomic configuration relative to the core molecule and are intended to be used as defined in the literature (IUP AC Recommendations 1996, Pure and Applied Chemistry,68:2193- > 2222 (1996)).

The term "tautomer" refers to each of two or more isomers of a compound that exist together in equilibrium and are readily interchangeable by the migration of atoms or groups within the molecule.

The term "halogen" or "halogen atom" or "halo" as used herein includes fluorine, chlorine, bromine and iodine atoms.

The term "5-membered heterocyclyl" denotes a mono-saturated or unsaturated group containing one or more heteroatoms selected from N and O.

Term "(C)_x-C_y) Alkyl "(where x and y are integers) represents a straight or branched alkyl group having from x to y carbon atoms. Thus, for example, when x is 1 and y is 6, the term includes methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl and n-hexyl.

Term "(C)_x-C_y) Alkylene "(wherein x and y are integers) represents C having a total of 2 unsaturated valences_x-C_yAlkyl residues, such as divalent methylene residues.

Expression "(C)_x-C_y) Haloalkyl "(wherein x and y are integers) represents" C "as defined above_x-C_yAn alkyl "group in which one or more hydrogen atoms are replaced by one or more halogen atoms, which may be the same or different.

Said "(C)_x-C_y) Examples of haloalkyl "groups may thus include halogenated, polyhalogenated and perhalogenated alkyl groups in which all hydrogen atoms are replaced by halogen atoms, such as trifluoromethyl.

Term "(C)_x-C_y) Cycloalkyl "(where x and y are integers) represents a saturated cyclic hydrocarbon group containing the indicated number of ring carbon atoms. Examples include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl.

The term "aryl" denotes a monocyclic ring system having 6 ring atoms, wherein the ring is aromatic. Examples of suitable aryl monocyclic systems include, for example, phenyl.

The term "heteroaryl" denotes a monocyclic or bicyclic aromatic group containing one or more heteroatoms selected from S, N and O, and includes groups having two such monocyclic rings, or one such monocyclic ring and one monocyclic aryl ring, which are fused by a common bond.

Term "(C)_x-C_y) Heterocycloalkyl "(wherein x and y are integers) denotes a saturated or partially unsaturated monocyclic ring (C)_x-C_y) Cycloalkyl in which at least one ring carbon atom is replaced by at least one heteroatom (e.g. N, S or O) or may carry an-oxo (═ O) substituent. The heterocyclic ringGroups may be further optionally substituted at available positions in the ring (i.e., on a carbon atom, or on a heteroatom available for substitution). Substitutions on carbon atoms include spiro disubstitution and substitution on 2 adjacent carbon atoms, in both cases thereby forming an additional fused 5-6 membered heterocyclic ring.

Term "(C)_x-C_y) Aminoalkyl "(wherein x and y are integers) denotes" (C) as defined above₁-C₆) An alkyl "group in which one or more hydrogen atoms are replaced by one or more amino groups.

Term "(C)_x-C_y) Hydroxyalkyl "(wherein x and y are integers) represents" (C) as defined above₁-C₆) An alkyl "group in which one or more hydrogen atoms are replaced by one or more hydroxyl (OH) groups.

Term "(C)_x-C_y) Alkoxy "or" (C)_x-C_y) Alkoxy "(where x and y are integers) represents a straight or branched chain hydrocarbon of the indicated number of carbons, which is linked to the rest of the molecule through an oxygen bridge.

A dash ("-") that is not between 2 letters or symbols is intended to represent a point of attachment for a substituent.

The carbonyl group is preferably represented herein as-C (O) -as a replacement for other commonly represented substituents such as-CO-, - (CO) -or-C (═ O) -.

In general, the groups in parentheses are pendant groups, not included in the chain, and parentheses are used to help disambiguate the linear formula when deemed useful; for example sulfonyl-SO₂-may also be represented by-S (O)₂To disambiguate, for example, with respect to sulfinyl-S (O) O-.

Whenever a basic amino or quaternary ammonium group is present in the compound of formula I, a physiologically acceptable anion may be present, selected from chloride, bromide, iodide, trifluoroacetate, formate, sulfate, phosphate, methanesulfonate, nitrate, maleate, acetate, citrate, fumarate, tartrate, oxalate, succinate, benzoate, p-toluenesulfonate, pamoate and napadisylate. Likewise, in the presence of acidic groups such as COOH groups, corresponding salts of physiological cations may also be present, including, for example, alkali or alkaline earth metal ions.

As indicated above, the present invention relates to a series of compounds represented by general formula (I), in particular as described hereinafter, which have inhibitory activity on the receptor LPA 2.

Advantageously, antagonist action at the receptor LPA2 may be effective in treating diseases where LPA receptors play a relevant role in pathogenesis, such as fibrosis and diseases, disorders and conditions caused by fibrosis.

Unlike similar compounds of the prior art, such as the compounds disclosed in Merck WO2012028243 and Amgen compounds, the compounds of formula (I) of the present invention are much more active on LPA 2.

Merck and Amgen compounds showed maximum potency expressed as half maximal Inhibitory Concentration (IC) to LPA2 at about 500nm₅₀)。

As indicated in the experimental part, in particular in table 2, the compounds of formula (I) of the present invention show a significant potency at less than about 500nm in their inhibitory activity on the receptor LPA2, demonstrating that they are able to antagonize the isoform (isoform) of the LPA2 receptor with greater potency than the compounds of the prior art, said isoform being involved in fibrosis and diseases caused by fibrosis.

Advantageously, the compounds of the present invention, characterized by very high potency, can be administered in humans at lower doses relative to the compounds of the prior art, thereby reducing the adverse events that typically occur when higher doses of drugs are administered.

Thus, the skilled person is particularly aware of the compounds of the present invention when studying suitable and effective compounds useful in the treatment of fibrosis, in particular idiopathic pulmonary fibrosis.

Thus, in one aspect, the invention relates to compounds of formula (I) as LPA2 antagonists

Wherein

R is H or selected from (C)₁-C₄) Alkyl, (C)₁-C₄) Haloalkyl and 5-6 membered heteroaryl, wherein each of said heteroaryl may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, halo, (C)₁-C₄) A haloalkyl group;

R₁is H or (C)₁-C₄) An alkyl group;

R₂is H or is selected from (C)₁-C₄) Alkyl, halo, (C)₁-C₄) Haloalkyl, (C)₃-C₈) A cycloalkyl group;

R₃is H or (C)₁-C₄) An alkyl group;

a is selected from 5-6 membered heteroaryl and aryl, wherein each of said heteroaryl and aryl may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, -C (O) R₁、-C(O)OR₁、-C(O)R₁、(C₁-C₄) Haloalkyl, halo, -NR_AC(O)R₁、-NR_AC(O)OR₁、-NR_AC(O)-(C₁-C₄) alkylene-OR₁、-NR_AC(O)R_C、-NR_AC(O)NR_AR_B、-N(C_1-C₄) alkylene-NR_AR_BAryl and heteroaryl, optionally substituted by one or more (C)₁-C₄) Alkyl and (C)₁-C₄) Haloalkyl is substituted, or

When a is aryl, it may be fused to a second saturated or unsaturated ring optionally containing one or more heteroatoms selected from N, O and S to form a bicyclic ring system, optionally substituted with one or more heteroatoms selected from-c (o) R₁、(C₁-C₄) Alkyl and oxo;

R_Cselected from heteroaryl, aryl, (C)₃-C₈) Cycloalkyl and (C)₄-C₈) Hetero compoundCycloalkyl, wherein said heteroaryl, aryl, heterocycloalkyl and cycloalkyl may optionally be substituted by one or more (C)₁-C₄) Alkyl and-C (O) OR₁Substitution;

R_Aand R_BIndependently at each occurrence is H or is selected from (C)₁-C₄) Alkyl, (C)₃-C₈) Cycloalkyl, (C)₁-C₆) Haloalkyl, halo, or

R_AAnd R_BMay form, together with the nitrogen atom to which they are attached, a 4-6 membered saturated heterocyclic ring system optionally containing another heteroatom selected from N, S or O, which may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, (C)₁-C₄) A halogenated alkyl group, a halogen-substituted,

provided that when R is methyl and A is aryl, the aryl is not substituted with one or more methyl groups and chlorine.

In a preferred embodiment, a is selected from 5-6 membered heteroaryl and aryl, wherein each of said heteroaryl and aryl may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, -C (O) R₁、-C(O)OR₁、-C(O)R₁、(C₁-C₄) Haloalkyl, halo, -NR_AC(O)R₁、-NR_AC(O)OR₁、-NR_AC(O)-(C₁-C₄) alkylene-OR₁、-NR_AC(O)R_C、-NR_AC(O)NR_AR_B、-N(C_1-C₄) alkylene-NR_AR_BAryl and heteroaryl (selected from iso-and iso-aryl)Oxazole, pyridine, thiazole,Oxazole, 1,2,4-Oxadiazole, 1,3,4-Diazoles and pyrazoles) optionally substituted with one or more (C)₁-C₄) Alkyl and (C)₁-C₄) Haloalkyl substitution; or

When a is aryl, it may be fused to a second saturated or unsaturated ring optionally containing one or more heteroatoms selected from N, O and S to form a bicyclic ring system, optionally substituted with one or more heteroatoms selected from-c (o) R₁、(C₁-C₄) Alkyl and oxo.

In a preferred embodiment, when a is a 5-6 membered heteroaryl, said 5-6 membered heteroaryl is selected from the group consisting of thiazole, thiophene and furan.

In a preferred embodiment, when R_CWhen it is heteroaryl, said heteroaryl is optionally substituted by one or more (C)₁-C₄) Alkyl and-C (O) OR₁Substituted heteroAnd (3) azole.

R is H or selected from (C)₁-C₄) Alkyl, (C)₁-C₄) Haloalkyl and 5-6 membered heteroaryl, wherein each of said heteroaryl may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, halo, (C)₁-C₄) A haloalkyl group;

R₁is H or (C)₁-C₄) An alkyl group;

R₂is H or is selected from (C)₁-C₄) Alkyl, halo, (C)₁-C₄) Haloalkyl, (C)₃-C₈) A cycloalkyl group;

R₃is (C)₁-C₄) An alkyl group;

a is selected from 5-6 membered heteroaryl and aryl, wherein each of said heteroaryl and aryl may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, -C (O) R₁、-C(O)OR₁、(C₁-C₄) Haloalkyl, halo, -NR_AC(O)R₁、-NR_AC(O)OR₁、-NR_AC(O)-(C₁-C₄) alkylene-OR₁、-NR_AC(O)R_CAryl and heteroaryl, optionally substituted by one or more (C)₁-C₄) Alkyl and (C)₁-C₄) Haloalkyl is substituted, or

When a is aryl, it may be fused to a second saturated or unsaturated ring optionally containing one or more heteroatoms selected from N, O and S to form a bicyclic ring system, optionally substituted with one or more heteroatoms selected from-c (o) R₁And (C)₁-C₄) Radical substitution of alkyl;

R_Cis selected from heteroaryl and (C)₄-C₈) Heterocycloalkyl, wherein said heteroaryl and heterocycloalkyl may optionally be substituted by one or more (C)₁-C₄) Alkyl and-C (O) OR₁The substitution is carried out by the following steps,

R_Ais H or (C)₁-C₄) An alkyl group, a carboxyl group,

provided that when R is methyl and A is aryl, the aryl is not substituted with one or more methyl groups and chlorine.

According to a preferred embodiment, the present invention relates to at least one compound listed in table 1 below and pharmaceutically acceptable salts thereof; those compounds were active against LPA2, as shown in table 2.

TABLE 1 list of preferred compounds of formula (I)

The compounds of the present invention can be prepared in a variety of ways known to those skilled in the art of organic synthesis. Those skilled in the art of organic synthesis will appreciate that the functional groups present on the molecule should be consistent with the proposed transformations. This sometimes requires modification of the order of the synthetic steps to obtain the desired compounds of the invention. The compounds of formula (I), including all compounds listed above, can generally be prepared according to the procedures outlined in the schemes shown below using generally known methods.

Scheme 1

The compound of formula (II) may be reacted with the nitrogen-based nucleophile of formula (III) in the presence of a suitable base, such as N, N-diisopropylethylamine in a suitable solvent, such as acetonitrile, to provide compound (IV) containing the Boc-protected amino group. Deprotection under well known procedures yields compound (V) which is finally reacted with a suitable sulfonyl chloride (VI) to give the compound of formula (I).

Examples 4-21,23-30 of the present invention can be prepared following the synthetic route outlined in scheme 1.

In another embodiment of the present invention, wherein R ═ CF₃And R is₂Compound (IXa) can be prepared from commercially available dihalothienopyrimidine (VI) according to scheme 2 as follows: formed by Grignard, converted to the corresponding iodide and subsequently triflated using a suitable reagent such as, for example, methyl 2, 2-difluoro-2- (fluorosulfonyl) acetate. Compound (VIII) is then converted to the final compound (IXa) following the synthetic sequence previously described in scheme 1. Examples 1 and 49 of the present invention can be prepared according to the synthetic route outlined in scheme 2.

Scheme 2

For where R ═ CF₃And R2 ≠ H, compound (IXb) can be synthesized as outlined in scheme 3. A compound of formula (X) (corresponding to intermediate 7-5 in WO 2013078765) can be reacted with a nitrogen-based nucleophile of formula (III) to provide compound (XI) containing a Boc-protected amino group. In the case of R2 ═ Cl, compound (XI) was converted directly to the final compound (IX) following the synthetic sequence outlined in scheme 1. For compounds where R2 ≠ H, in the presence of a suitable palladium catalyst such as [1, 1' -bis (diphenylphosphino) ferrocene]The Cl intermediate (XI) was subjected to Suzuki coupling in the presence of palladium (II) dichloride to introduce the appropriate R2 substituent and then the same synthetic sequence as outlined in scheme 1 was performed to give the final compound (IXb). Examples 32 and 34 of the present invention can be prepared according to the synthetic route outlined in scheme 3.

Scheme 3

In another embodiment of the present invention, wherein R ═ aryl, heteroaryl, intermediate (II) is obtained following the synthetic route outlined in scheme 4. Dihalothienopyrimidine (VI) may be converted to methylsulfanyl) -derivative (XIII), which undergoes Suzuki coupling with commercially available boronic acid to provide intermediate (XIV). Subsequent reaction of compound (XI) with sulfuryl dichloride affords intermediate (II), which is converted to the final compound (I) according to the synthetic sequence previously described in scheme 1. Examples 2, 39, 40, 42-45, 47, 53-54 of the present invention can be prepared according to the synthetic route outlined in scheme 4.

Scheme 4

Alternatively, according to scheme 5, for compounds wherein R ═ aryl, heteroaryl, the Intermediate (IV) can be obtained as follows: nucleophilic substitution on compound (VI) with a nitrogen-based nucleophile of formula (III) followed by Suzuki coupling with a commercially available boronic acid. Compound (IV) is then converted to the final compound (I) following the synthetic sequence previously described in scheme 1.

Scheme 5

In another embodiment of the present invention, compound (II) can be synthesized as reported in scheme 6, wherein R2 ≠ H.

Scheme 6

The commercially available compound (XVI) is cyclized with the appropriate nitrile (XVII) and subsequently with a suitable chlorinating agent such as POCl₃Chlorination to provide intermediate (II). Compound (II) is then converted to the final compound (I) following the synthetic sequence previously described in scheme 1. Examples 22, 31, 35-38 of the present invention can be prepared according to the synthetic route outlined in scheme 6.

For compounds where R2 ≠ H and R ═ aryl, heteroaryl, the synthetic pathway outlined in scheme 7 gave the Intermediate (IV).

Scheme 7

The commercially available compound (XIX) is cyclized with the appropriate nitrile (XIV), followed by a suitable chlorinating agent such as POCl₃Chlorination provides intermediate (XXI). Nucleophilic substitution with a nitrogen-based nucleophile of formula (III) followed by Suzuki coupling with a commercially available boronic acid gives Intermediate (IV). Compound (IV) is then converted to the final compound (I) according to the synthetic sequence previously described in scheme 1). Examples 3, 46 and 48 of the present invention can be prepared following the synthetic route outlined in scheme 7.

In another embodiment of the invention, wherein a is N-acylated aminothiazole, R4 and R5 ═ H, CH₃Compound (XXVI) can be obtained according to scheme 8. Intermediate (V) is reacted with N-acetylthiazolesulfonyl chloride (XXII), followed by deacetylation under acid conditions and finally acetylation with the appropriate acid chloride in the presence of a base such as N, N-dimethyl-4-pyridylamine to give the final compound (XXVI). Examples 33, 41, 50-52, 55 of the present invention can be prepared according to the synthetic route outlined in scheme 8.

Scheme 8

It has surprisingly been found that the compounds of formula (I) of the present invention effectively inhibit the receptor LPA 2. Advantageously, inhibition of LPA2 may result in effective treatment of diseases or conditions in which LPA receptors are involved.

In particular in this regard, it has now been found that the compounds of formula (I) of the present invention have antagonist drug efficacy, expressed as the half maximal Inhibitory Concentration (IC) to LPA2 of less than or equal to 1000nM₅₀) As shown in the experimental part of the present invention.

Preferably, the compounds of the invention have an IC for LPA2 of less than or equal to 100nM₅₀。

More preferably, the compounds of the invention have an IC for LPA2 of less than or equal to 10nM₅₀。

In one aspect, the present invention relates to a compound of formula (I) for use as a medicament.

In a preferred embodiment, the present invention relates to compounds of formula (I) for use in the treatment of disorders related to the mechanisms of the LPA receptor.

In another embodiment, the present invention relates to compounds of formula (I) for use in the treatment of a disease, disorder or condition associated with dysregulation of lysophosphatidic acid receptor 2(LPA 2).

In one embodiment, the present invention relates to compounds of formula (I) useful for the prevention and/or treatment of fibrosis and/or diseases, disorders or conditions involving fibrosis.

The term "fibrosis" or "fibrotic disorder" as used herein denotes a condition associated with abnormal accumulation of cells and/or fibronectin and/or collagen and/or increased fibroblast recruitment, and includes, but is not limited to, fibrosis of various organs or tissues such as heart, kidney, liver, joint, lung, pleural tissue, peritoneal tissue, skin, cornea, retina, musculoskeletal and digestive tract.

Preferably, the compounds of formula (I) of the present invention are useful for the treatment and/or prevention of fibrosis such as pulmonary fibrosis, Idiopathic Pulmonary Fibrosis (IPF), liver fibrosis, kidney fibrosis, eye fibrosis, cardiac fibrosis, arterial fibrosis and systemic sclerosis.

More preferably, the compounds of formula (I) of the present invention are useful for the treatment of Idiopathic Pulmonary Fibrosis (IPF).

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

In one aspect, the present invention relates to the use of a compound of formula (I) for the manufacture of a medicament for the treatment of disorders related to the mechanisms of the LPA receptors.

In another aspect, the present invention relates to a method for the prevention and/or treatment of disorders or conditions associated with dysregulation of lysophosphatidic acid receptor 2(LPA2), comprising administering to a patient in need of such treatment a therapeutically effective amount of a compound of formula (I).

In another aspect, the present invention relates to the use of a compound of formula (I) according to the present invention, or a pharmaceutically acceptable salt thereof, for the treatment of disorders related to the mechanisms of the LPA receptor.

In another aspect, the present invention relates to the use of a compound of formula (I) for the treatment of a disease, disorder or condition associated with dysregulation of receptor 2(LPA 2).

As used herein, "safe and effective amount" with respect to a compound of formula (I) or a pharmaceutically acceptable salt or other pharmaceutically active agent thereof refers to the amount of such compound: it is sufficient to treat the condition of the patient, but low enough to avoid serious side effects, although it can be routinely determined by the skilled artisan.

The compound of formula (I) or a pharmaceutically acceptable salt thereof may be administered once or according to a dosing regimen in which several doses are administered at varying intervals over a given period of time. Typical daily dosages may vary depending upon the route of administration selected.

The invention also relates to pharmaceutical compositions comprising a compound of formula (I) in admixture with at least one or more pharmaceutically acceptable carriers or excipients.

In one embodiment, the invention relates to pharmaceutical compositions of compounds of formula (I), such as those described in Remington's pharmaceutical Sciences Handbook, XVII edition, Mack pub.

Administration of the compounds of the present invention and pharmaceutical compositions thereof may be accomplished in accordance with the patient's needs, for example, orally, nasally, parenterally (subcutaneously, intravenously, intramuscularly, intrasternally and by infusion) and by inhalation.

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

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

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

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

The compounds of the present invention may be administered alone or in combination with various pharmaceutically acceptable carriers, diluents (such as sucrose, mannitol, lactose, starch) and known excipients including suspending agents, solubilizers, buffers, binders, disintegrants, preservatives, colorants, flavorants, lubricants and the like.

In another embodiment, the pharmaceutical compositions comprising the compounds of formula (I) are liquid oral dosage forms such as aqueous and non-aqueous solutions, emulsions, suspensions, syrups, and elixirs. Such liquid dosage forms may also contain suitable known inert diluents such as water and suitable known excipients such as preservatives, wetting agents, sweeteners, flavoring agents, and agents for emulsifying and/or suspending the compounds of the present invention.

In another embodiment, the pharmaceutical composition comprising a compound of formula (I) is an inhalable formulation such as an inhalable powder, a propellant-containing metered dose aerosol or a propellant-free inhalable formulation.

For administration as a dry powder, single-or multidose inhalers known from the prior art can be used. In this case, the powder may be filled in gelatin, plastic or other capsules, cartridges (cartridges) or blister packs or in a reservoir.

A diluent or carrier that is chemically inert to the compounds of the invention, such as lactose or any other additive suitable for improving the respirable fraction, may be added to the powdered compounds of the invention.

Inhalation aerosols comprising a propellant gas, such as a hydrofluoroalkane, may comprise the compounds of the invention in solution or in dispersed form. The propellant-driven formulation may also comprise other ingredients, such as co-solvents, stabilizers or optionally other excipients.

Propellant-free inhalable formulations comprising the compounds of the invention may be in the form of solutions or suspensions in aqueous, alcoholic or hydroalcoholic media, and they may be delivered by spray or ultrasonic nebulizers or by soft mist nebulizers as known in the art.

The compounds of the present invention may be administered as the sole active agent or in combination with other pharmaceutically active ingredients.

The dosage of the compounds of the present invention will depend on a variety of factors including the particular disease to be treated, the severity of the symptoms, the route of administration, and the like.

The invention also relates to a device comprising a pharmaceutical composition comprising a compound of formula (I) according to the invention, in the form of a single-dose or multi-dose dry powder inhaler or a metered dose inhaler.

All preferred groups or embodiments described above for the compounds of the formula I can be combined with one another and apply mutatis mutandis.

The various aspects of the invention described herein are illustrated by the following examples, which are not intended to limit the invention in any way.

Preparation of intermediates and examples

The chemical Name of the compound was generated using Structure To Name Enterprise 10.0Cambridge Software.

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

Abbreviation-meaning

AcOEt ═ ethyl acetate

CsCO₃Cesium carbonate (MCH)

CyHex ═ cyclohexane

DCM ═ dichloromethane

DMF ═ dimethylformamide

DMSO ═ dimethyl sulfoxide

h is hour

HCl ═ hydrochloric acid

HCOOH ═ formic acid

H₂O is water

K₃PO₄Phosphoric acid tripotassium salt

LC-MS (liquid chromatography-mass spectrometry)

MeCN ═ acetonitrile

MW ═ microwave

NaCl sodium chloride

Na₂CO₃Sodium carbonate (sodium bicarbonate)

NaHCO₃Sodium bicarbonate

Na₂SO₃Sodium sulfite

Na₂SO₄Sodium ═ sulfate

NH₄Cl ═ ammonium chloride

NH₃Ammonia (II) is

N₂Is nitrogen

Pd(dppf)Cl₂1, 1' -bis (diphenylphosphino) ferrocene]Palladium dichloride (II)

Pd(PPh₃)₄Tetra (triphenylphosphine) -palladium (0)

POCl₃Phosphorus oxychloride (V)

r.t. room temperature

THF ═ tetrahydrofuran

UPLC (ultra performance liquid chromatography)

General experimental details

Analytical method

Apparatus, materials and methods for analysis

¹ H-NMR

On a Varian MR-400 spectrometer operating at 400MHz (proton frequency)¹H-NMR spectrum, which is provided with: self-shielded Z-gradient coil 5mm 1H/nX broadband probe for reverse detection, deuterium digitally locked channel unit, quadrature digital detection unit with transmitter offset frequency shift, either on Agilent VNMRS-500 or on Bruker Avance 400 or on Bruker Avance 300 spectrometer¹H-NMR spectrum. Chemical shifts are reported as 6 ppm values relative to Trimethylsilane (TMS) as an internal standard. Coupling constants (J values) are given in hertz (Hz) and multiplicities are reported using the following abbreviations (s ═ singlet, d ═ doublet, t ═ triplet, q ═ quartet, m ═ multiplet, br.s ═ broad singlet, and nd ═ undetermined).

LC/UV/MS

Estimating LC/MS retention time is subject to+0.5min of experimental error. LCMS can be recorded under the following conditions: diode array DAD chromatographic traces, mass chromatograms and mass spectrograms can be taken from the following systems: micromass ZQ with ES ionization mode operating in positive and/or negative electron injection^TMOr Waters SQD Single quadrupole Mass spectrometer coupled UPLC/PDA/MS acquisition^TMSystems, and/or methods, for operating with ZQ in positive and/or negative ES ionization mode^TMA single quadrupole coupled Fractionlynx system used in analytical mode, or Waters Alliance e2695 with photodiode detector 2998 coupled to Column Oven and Mass Spectrometer ZQ operating in positive and/or negative ES ionization mode. Quality control method of operation used under low pH conditions or under high pH conditions:

method 1, low pH conditions, column: Acquity CSH C182.1x50mm 1.7 μm, column temperature 40 ℃; mobile phase solvent A was milliQ water + 0.1% HCOOH, and mobile phase solvent B was MeCN + 0.1% HCOOH. The flow rate was 1 mL/min. The gradient is t 0min 97% A3% B, t 1.5min 0.1% a 99.9% B, t 1.9min 0.1% a 99.9% B and t 2min 97% A3% B. The ultraviolet detection range is 210-350nm and the ES +/ES-range is 100-1500 AMU.

Method 2, high pH conditions, column: Acquity Kinetex 1.7 μm EVO C18100A, 2.1X50mm, column temperature 40 ℃; mobile phase solvent A was 10mM NH adjusted to pH 10 with ammonia₄HCO₃The aqueous solution, mobile phase solvent B was MeCN. The flow rate was 1 mL/min. The gradient table is t 0min 97% A3% B, t 1.5min 0.1% a 99.9% B, t 1.9min 0.1% a 99.9% B and t 2min 97% A3% B. The ultraviolet detection range is 210-350nm and the ES +/ES-range is 100-1500 AMU.

Method 3, low pH condition, column, Acquity CSH C182.1x50mm 1.7 μm, column temperature 40 deg.C; mobile phase solvent A was milliQ water/MeCN 95:5+ 0.05% HCOOH, and mobile phase solvent B was MeCN/milliQ water 95:5+ 0.05% HCOOH. The flow rate was 1 mL/min. The gradient is t 0min 99% A1% B, t 1.5min 0.1% a 99.9% B, t 1.9min 0.1% a 99.9% B and t 2min 99% A1% B. The ultraviolet detection range is 210-400nm and the ES +/ES-range is 100-1200 AMU.

Method 4, low pH conditions, column Phenomenex Gemini-NX C18,150x2.0mm,3 μm, Gemini-NX C18 with safety protector (guard), 4x2.0mm,3 μm, column temperature 25 deg.C; mobile phase solvent a was water filtered through a 0.22 μm nylon filter + 0.1% formic acid, and mobile phase solvent B was acetonitrile filtered through a 0.22 μm nylon filter + 0.1% formic acid. The flow rate was 0.2 mL/min. The gradient table is t 0min 95% A5% B, t 10min 20% a 80% B, t 30min 20% a 80B. The UV detection lambda is 210nm and the ES +/ES-range is 50-900 Da.

Method 5, low pH conditions, column of Acquity CSH C182.1x50mm 1.7 μm at 40 deg.C; mobile phase solvent A was milliQ water/MeCN 95:5+ 0.05% HCOOH, and mobile phase solvent B was MeCN/milliQ water 95:5+ 0.05% HCOOH. The flow rate was 1 mL/min. The gradient table is t 0min 99% A1% B, t 3.5min 0.1% a 99.9% B, t 3.9min 0.1% a 99.9% B and t 4min 99% A1% B. The ultraviolet detection range is 210-400nm and the ES +/ES-range is 100-1200 AMU.

Example 1

Methyl N- [ 4-methyl-5- ({4- [ (2S) -2- { [7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] carbamate

Step 1: preparation of 4-chloro-7-iodothieno [3,2-d ] pyrimidine (intermediate 1)

Reacting 7-bromo-4-chlorothieno [3,2-d]Pyrimidine (1g,4mmol) was dissolved in THF (10mL), cooled at 0 deg.C and a 2.0M solution of isopropyl magnesium chloride in THF (3mL,6mmol) was added dropwise. After stirring at 0 ℃ for 30min, the mixture was treated dropwise with a solution of iodine (1.53g,6mmol) in THF (20 mL). After 1 hour at 0 ℃ the reaction was quenched by addition of saturated NH₄Aqueous Cl was quenched and partitioned between AcOEt and water. The organic phase was saturated with Na₂SO₃And (4) washing the solution. Subjecting the organic layer to Na₂SO₄Dried, filtered and concentrated under reduced pressure to give a crudePurified by flash chromatography eluting with MeCN (from 0% to 40%) in water to provide the title compound as a brown solid (850mg,2.87mmol, 71% yield).

LC-MS (ESI) M/z (M +1):297 (method 1)

¹H NMR(400MHz,DMSO-d6)δppm 9.16(s,1H),8.84(s,1H)

Step 2: preparation of 4-chloro-7- (trifluoromethyl) thieno [3,2-d ] pyrimidine (intermediate 2)

Copper (I) iodide (576.28mg,3mmol) was dissolved in DMF (15mL) and methyl 2, 2-difluoro-2- (fluorosulfonyl) acetate (0.73mL,5.7mmol) was added. The resulting solution was stirred at room temperature for 5 minutes. Intermediate 1(850mg,2.87mmol) was added and the reaction was stirred at 80 ℃ overnight and then at room temperature for 3 days. The mixture was filtered and the solid was washed with ethyl acetate. The filtrate was washed with a saturated solution of NaCl (3 times). Passing the organic phase over Na₂SO₄Dried, filtered and evaporated under reduced pressure. The crude was purified by flash chromatography eluting with AcOEt in cyclohexane (from 0% to 10%) to give the title compound as a yellowish solid (459mg,1.9mmol, 67% yield).

LC-MS (ESI): M/z (M +1):239 (method 1)

¹H NMR(400MHz,DMSO-d6)δppm 9.29(s,1H),9.21(s,1H)

And step 3: preparation of tert-butyl 4- [ (2S) -2- [ [7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino ] propyl ] piperazine-1-carboxylate (intermediate 3)

To a solution of intermediate 2(459mg,1.9mmol) in MeCN (15mL) was added 4- [ (2S) -2-aminopropyl)]Piperazine-1-carboxylic acid tert-butyl ester (468mg,1.9mmol) followed by N, N-diisopropylethylamine (0.37mL,2.12mmol) was added. The solution was brought to 40 deg.CHeat overnight. The mixture was diluted with water and AcOEt and the aqueous phase was extracted with AcOEt (2 times). The combined organic layers were passed over Na₂SO₄Dried, filtered and concentrated under reduced pressure. The crude was purified by flash chromatography eluting with AcOEt in cyclohexane (from 0% to 60%) to provide the title compound as a white solid (450mg,1mmol, 52% yield).

LC-MS (ESI): M/z (M +1):446 (method 1)

¹H NMR(400MHz,DMSO-d₆)δppm 8.78(s,1H),8.50(s,1H),7.94(d,J＝8.1Hz,1H),4.61(p,J＝6.9Hz,1H),3.15-3.29(m,4H),2.53-2.59(m,1H),2.34-2.40(m,5H),1.38(s,9H),1.22(d,J＝6.6Hz,3H)

And 4, step 4: preparation of N- [ (2S) -1-piperazin-1-ylpropan-2-yl ] -7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-amine (intermediate 4)

To a solution of intermediate 3(450mg,1mmol) in DCM (12mL) was added 2,2, 2-trifluoroacetic acid (1.16mL,15 mmol). The reaction was stirred at rt overnight. The mixture was then concentrated under reduced pressure and the residue was purified using an SCX column (cartridge) using 2M NH₃A solution in methanol was eluted to give the title compound as a yellow solid (340mg,0.98mmol, 97% yield).

LC-MS (ESI): M/z (M +1):346 (method 2)

¹H NMR(400MHz,DMSO-d₆)δppm 8.78(d,J＝1.1Hz,1H),8.49(s,1H),7.90(d,J＝8.1Hz,1H),4.60(p,J＝6.7Hz,1H),2.56-2.70(m,4H),2.46-2.48(m,2H),2.20-2.41(m,5H),1.21(d,J＝6.6Hz,3H)

And 5: preparation of methyl N- [ 4-methyl-5- ({4- [ (2S) -2- { [7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] carbamate (example 1).

To a solution of intermediate 4(340mg,0.98mmol) in pyridine/DCM 3:2(5mL) was added methyl N- [5- (chlorosulfonyl) -4-methyl-1, 3-thiazol-2-yl ] carbamate (94.05mg,0.350mmol), and the reaction was stirred at room temperature for 12 hours. The solvent was removed under vacuum and the crude product was purified by flash chromatography, eluting with from 0% to 25% AcOEt in cyclohexane, to provide the title compound as a white solid (22.6mg,0.039mmol, 13.47% yield).

LC-MS (ESI): M/z (M +1):580 (method 1)

¹H NMR(400MHz,DMSO-d₆)δppm 12.37(br s,1H),8.76(s,1H),8.48(s,1H),7.91(d,J＝8.0Hz,1H),4.53(dt,J＝13.9,7.1Hz,1H),3.73(s,3H),2.97(br s,4H),2.52-2.60(m,5H),2.41(s,3H),2.32-2.38(m,1H),1.18(d,J＝6.6Hz,3H)

The examples in the table below were prepared from commercially available reagents using a similar method to example 1.

Example 2

N- [5- ({4- [ (2S) -2- { [7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) thieno [3,2-d]Pyrimidin-4-yl]Amino } propyl group]Piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl]Carbamic acid methyl ester

Step 1: preparation of 7-bromo-4- (methylsulfanyl) thieno [3,2-d ] pyrimidine (intermediate 5)

To a solution of 7-bromo-4-chlorothio [3,2-d ] pyrimidine (100mg,0.4mmol) in THF (1.5mL) was added sodium thiomethoxide (70mg,1mmol) at 0 deg.C. The mixture was stirred at room temperature overnight. Ice water was added and the resulting solid was filtered and dried to provide the title compound as a white solid (85mg,0.33mmol, 83% yield).

LC-MS (ESI): M/z (M +1):262.9 (method 1)

¹H NMR(400MHz,DMSO-d6)δppm 9.10(s,1H),8.59(s,1H),2.78(s,3H)

Step 2: preparation of 3, 5-dimethyl-4- (4-methylsulfanyl thieno [3, 2-d)]Pyrimidin-7-yl) -1,2-Azole (intermediate 6)

To intermediate 5(85mg,0.33mmol) and 3, 5-dimethyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) -1,2-To a suspension of oxazole (128mg,0.5mmol) in water (0.7mL) was added cesium carbonate (249mg,0.66 mmol). With N₂After degassing, Pd (PPh) was added₃)₄(44mg,0.04mmol) and the reaction was heated at 100 ℃ for 16 h. After cooling, AcOEt was added and the crude mixture was washed with a saturated solution of NaCl. The organic layer was concentrated under reduced pressure. The crude was purified by flash chromatography eluting with from 0% to 20% AcOEt in cyclohexane to give the title compound (26.5mg,0.096mmol, 29% yield).

LC-MS (ESI): M/z (M +1):278 (method 1)

And step 3: preparation of 4- (4-chlorothieno [3, 2-d)]Pyrimidin-7-yl) -3, 5-dimethyl-1, 2-Azole (intermediate 7)

To a solution of intermediate 6(226.5mg,0.82mmol) in DCM (2.5mL)Sulfuryl dichloride (0.26mL,3.27mmol) was added to the solution and the reaction was stirred at 25 ℃ for 15 min. Adding saturated NaHCO successively₃Solution and DCM, then the organic layer was separated and concentrated under reduced pressure to give the title compound (323mg, crude) which was used in the next step without further purification.

LC-MS (ESI) M/z (M +1) 266.3 (method 1)

And 4, step 4: preparation of 4- [ (2S) -2- [ [7- (3, 5-dimethyl-1, 2-Oxazol-4-yl) thieno [3,2-d]Pyrimidin-4-yl]Amino group]Propyl radical]Piperazine-1-carboxylic acid tert-butyl ester (intermediate 8)

Following the procedure used for the synthesis of intermediate 4, starting from 4- (4-chlorothieno [3, 2-d)]Pyrimidin-7-yl) -3, 5-dimethyl-1, 2-The title compound was prepared starting from oxazole (intermediate 7,323mg, crude) to provide the title compound (247mg,0.52mmol, 64% yield).

LC-MS (ESI): M/z (M +1):472.9 (method 2)

And 5: preparation of 7- (3, 5-dimethyl-1, 2-Oxazol-4-yl) -N- [ (2S) -1-piperazin-1-ylpropan-2-yl]Thieno [3,2-d]Pyrimidine-4-amine hydrochloride (intermediate 9)

To 4- [ (2S) -2- [ [7- (3, 5-dimethyl-1, 2-Oxazol-4-yl) thieno [3,2-d]Pyrimidin-4-yl]Amino group]Propyl radical]Piperazine-1-carboxylic acid tert-butyl esterButyl ester (intermediate 8,247mg,0.52mmol) in 1, 4-bisTo a solution in alkane (3mL), 4M HCl in 1, 4-bisSolution in alkane (0.65mL,2.61 mmol). The reaction was stirred at rt for 3 h. The mixture was then concentrated under reduced pressure to give the title compound (300mg, crude).

LC-MS (ESI) M/z (M +1):373.4 (method 2)

And 5: preparation of N- [5- ({4- [ (2S) -2- { [7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) thieno [3,2-d]Pyrimidin-4-yl]Amino } propyl group]Piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl]Carbamic acid methyl ester (example 2)

Following the procedure used for the synthesis of example 1, starting from 7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) -N- [ (2S) -1-piperazin-1-ylpropan-2-yl]Thieno [3,2-d]Pyrimidin-4-amine hydrochloride (intermediate 9,65mg,0.158mmol) and N- [5- (chlorosulfonyl) -4-methyl-1, 3-thiazol-2-yl]Methyl carbamate (43.46mg,0.160mmol) the title compound was prepared starting from methyl carbamate to provide the title compound as a white solid (26.5mg,0.044mmol, 28% yield).

LC-MS (ESI) M/z (M +1):607.08 (method 2)

¹H NMR(400MHz,DMSO-d₆)δppm 12.33(br s,1H),8.42(s,1H),8.13(s,1H),7.67(d,J＝8.1Hz,1H),4.44-4.62(m,1H),3.75(s,3H),2.99(br s,4H),2.56(br s,5H),2.15-2.46(m,9H),1.18(d,J＝6.4Hz,3H)

The examples in the table below were prepared from commercially available reagents using a similar method to example 2.

Example 3

2-cyclopropyl-7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) -N- [ (2S) -1- (4- { [5- (3-methyl-1, 2-)Azol-5-yl) thiophen-2-yl]Sulfonyl-piperazin-1-yl) -propan-2-yl]Thieno [3,2-d]Pyrimidin-4-amines

Step 1: preparation of 7-bromo-2-cyclopropyl-3H-thieno [3,2-d ] pyrimidin-4-one (intermediate 10)

To a solution of methyl 3-amino-4-bromothiophene-2-carboxylate (1.18g,5mmol) in cyclopropanecarbonitrile (11mL,165.39mmol) was added 2N HCl (2.5mL,5 mmol). The mixture was heated at 130 ℃ for 21 h periods under MW irradiation. Adding saturated NaHCO₃The solution was extracted 3 times with AcOEt; the combined organic layers were passed over Na₂SO₄Dried, filtered and evaporated under vacuum. The crude was purified by flash chromatography eluting with cyclohexane/AcOEt from 9/1 to 1/1 to provide the title compound as a light yellow solid (163mg,0.6mmol, 12% yield).

LC-MS (ESI) M/z (M +1):273 (method 1)

¹H NMR(400MHz,DMSO-d₆)δppm 12.83(br.s.,1H),8.30(s,1H),1.97-2.08(m,1H),1.02-1.14(m,4H)

Step 2: preparation of 7-bromo-4-chloro-2-cyclopropylthieno [3,2-d ] pyrimidine (intermediate 11)

Intermediate 10(163mg,0.6mmol) in POCl₃The solution in (10mL) was stirred at 100 ℃ overnight. Evaporation of POCl₃Then adding H₂O (5mL) and NaHCO₃(5mL), and the mixture was added. The combined organic layers were passed over Na₂SO₄Dried, filtered and evaporated to give the title compound (150mg,0.52mmol, 73% yield), which was used in the next step without further purification.

LC-MS (ESI): M/z (M +1):290.9 (method 1)

¹H NMR(400MHz,DMSO-d₆)δppm 8.71(s,1H),2.31-2.42(m,1H),1.06-1.21(m,4H)

And step 3: preparation of tert-butyl 4- [ (2S) -2- [ (7-bromo-2-cyclopropylthieno [3,2-d ] pyrimidin-4-yl) amino ] propyl ] piperazine-1-carboxylate (intermediate 12)

The title compound was prepared following the procedure used for the synthesis of intermediate 3, starting from 7-bromo-4-chloro-2-cyclopropylthieno [3,2-d ] pyrimidine (intermediate 11,150mg,0.52mmol) to provide the title compound as a white solid (248mg,0.5mmol, 96.5% yield).

LC-MS (ESI) M/z (M +1) 498.1 (method 1)

¹H NMR(400MHz,DMSO-d6)δppm 8.21(s,1H),7.62(d,J＝7.92Hz,1H),4.40-4.59(m,1H),4.04(q,J＝7.12Hz,1H),3.19-3.29(m,4H),2.35-2.46(m,4H),2.30(dd,J＝12.21,7.37Hz,1H),2.02-2.10(m,1H),1.39(s,9H),1.19-1.27(m,3H),0.88-1.08(m,4H)

And 4, step 4: preparation of 4- [ (2S) -2- [ [ 2-cyclopropyl-7- (3, 5-dimethyl-1, 2-Oxazol-4-yl) thieno [3,2-d]Pyrimidin-4-yl]Amino group]Propyl radical]Piperazine-1-carboxylic acid tert-butyl ester (intermediate 13)

To a solution of intermediate 12(248mg,0.5mmol) in THF (5mL) and water (2mL) was added 3, 5-dimethyl-4- (4,4,5, 5-tetramethyl-1, 3, 2-dioxaborolan-2-yl) iso-pentaneOxazole (446mg,2mmol), Pd (dppf) Cl₂(36.67mg,0.05mmol) and K₃PO₄(215mg,1 mmol). The mixture was stirred at 80 ℃ overnight. Brine was added and the mixture was extracted 3 times with AcOEt. The combined organic layers were passed over Na₂SO₄Dried, filtered and evaporated to dryness to give the crude product, which was purified by flash column chromatography eluting with cyclohexane/AcOEt from 6/4 to 2/8 to give the title compound as a yellow oil (430mg,0.84mmol, 84% yield).

LC-MS (ESI): M/z (M +1):513.2 (method 1)

¹H NMR(400MHz,DMSO-d₆)δppm 8.06(s,1H),7.92(s,1H),7.50(d,J＝7.70Hz,1H),4.45-4.62(m,1H),3.92(s,6H),3.27(t,J＝4.62Hz,3H),2.36-2.45(m,6H),2.32(dd,J＝12.21,7.37Hz,1H),2.22(s,3H),1.95-2.03(m,2H),1.39(s,8H),1.21-1.27(m,3H),1.14-1.21(m,3H),1.08(s,36H),0.86-0.97(m,4H)

And 5: preparation of 2-cyclopropyl-7- (3, 5-dimethyl-1, 2-Oxazol-4-yl) -N- [ (2S) -1-piperazin-1-ylpropan-2-yl]Thieno [3,2-d]Pyrimidine-4-amine hydrochloride (intermediate 14)

Following the procedure used for the synthesis of intermediate 9, starting from 4- [ (2S) -2- [ [ 2-cyclopropyl-7- (3, 5-dimethyl-1, 2-Oxazol-4-yl) thieno [3,2-d]Pyrimidin-4-yl]Amino group]Propyl radical]Tert-butyl piperazine-1-carboxylate (intermediate 13,430mg,0.84mmol) the title compound was prepared starting from to provide the title compound as a light yellow solid (394mg,0.81mmol, 97% yield).

LC-MS (ESI): M/z (M +1):413.1 (method 2)

¹H NMR(400MHz,DMSO-d₆)δppm 9.47-9.78(m,1H),8.20-8.46(m,1H),4.78-4.99(m,1H),3.67-4.03(m,11H),3.31-3.52(m,4H),2.35(s,3H),2.16(s,2H),1.10-1.35(m,4H),1.08(s,10H)

Step 6: preparation of 2-cyclopropyl-7- (3, 5-dimethyl-1, 2-Oxazol-4-yl) -N- [ (2S) -1- (4- { [5- (3-methyl-1, 2-)Azol-5-yl) thiophen-2-yl]Sulfonyl } piperazin-1-yl) propan-2-yl]Thieno [3,2-d]Pyrimidine-4-amines (example 3)

Following the procedure used for the synthesis of example 1, starting from 2-cyclopropyl-7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) -N- [ (2S) -1-piperazin-1-ylpropan-2-yl]Thieno [3,2-d]Pyrimidin-4-amine hydrochloride(intermediate 14,394mg,0.81mmol) and 5- (3-methyl-1, 2-)Oxazol-5-yl) thiophene-2-sulfonyl chloride (235mg,0.89mmol) the title compound was prepared to provide the title compound as a white solid (77mg,0.12mmol, 15% yield).

LC-MS (ESI) M/z (M +1):640 (method 2)

¹H NMR(400MHz,DMSO-d₆)δppm 8.02(s,1H),7.77(d,J＝4.0Hz,1H),7.70(d,J＝4.0Hz,1H),7.46(d,J＝7.8Hz,1H),6.97(s,1H),4.38-4.53(m,1H),2.98(br s,4H),2.56-2.63(m,4H),2.48-2.63(m,1H),2.35(s,3H),2.29(s,3H),2.26-2.36(m,1H),2.18(s,3H),1.93-2.02(m,1H),1.17(d,J＝6.6Hz,3H),0.83-0.92(m,4H)

The examples in the table below were prepared from commercially available reagents using a similar method to example 3.

Example 4

5- ({4- [ (2S) -2- ({ 7-methylthiophen-o [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) thiophene-2-carboxylic acid methyl ester

Step 1: preparation of tert-butyl 4- [ (2S) -2- ({ 7-methylthio [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazine-1-carboxylate (intermediate 15)

The title compound was prepared following the procedure used for the synthesis of intermediate 3, starting from commercially available 4-chloro-7-methylthioeno [3,2-d ] pyrimidine (1.0g,5.42mmol) to provide the title compound (1.61g,4.11mmol, 76% yield) as a white (whitenish) solid.

LC-MS (ESI) M/z (M +1):392.3 (method 2)

Step 2: preparation of 7-methyl-N- [ (2S) -1-piperazin-1-ylpropan-2-yl ] thieno [3,2-d ] pyrimidin-4-amine (intermediate 16)

The title compound was prepared following the procedure used for the synthesis of intermediate 4 starting from tert-butyl 4- [ (2S) -2- ({ 7-methylthioeno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazine-1-carboxylate (intermediate 15,1.61g,4.11mmol) to afford the title compound as a yellowish solid (1.12g,3.84mmol, 98% yield).

LC-MS (ESI) M/z (M +1):292.2 (method 2)

And step 3: preparation of methyl 5- ({4- [ (2S) -2- ({ 7-methylthio [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) thiophene-2-carboxylate

(example 4)

To (7-methyl-N- [ (2S) -1-piperazin-1-ylpropan-2-yl)]Thieno [3,2-d]To a solution of pyrimidin-4-amine trihydrochloride (intermediate 16,50mg,0.125mmol) in dry pyridine (0.7ml) was added methyl 5- (chlorosulfonyl) thiophene-2-carboxylate (30mg,0.125mmol) and the solution was sonicated for 5 min. After evaporation of the pyridine, the residue was treated with heptane (35ml) and the solvent was evaporated in vacuo. The crude was taken up in saturated NaHCO₃Partition between solution (10mL) and EtOAc (30mL) and pass the organic layer over Na₂SO₄Dried and evaporated to dryness to give the title compound as a yellow solid.

LC-MS (ESI) M/z (M +1):496.18 (method 3)

¹H NMR (400MHz, acetone-d₆)δppm 8.47(s,1H)7.83(d,J＝3.91Hz,1H)7.57(d,J＝4.03Hz,1H)7.52(d,J＝1.22Hz,1H)6.42(br d,J＝7.09Hz,1H)4.56-4.67(m,1H)3.92(s,3H)3.03(br s,4H)2.59-2.73(m,5H)2.47(dd,J＝12.53,6.42Hz,1H)2.35(d,J＝1.10Hz,3H)1.27(d,J＝6.48Hz,3H)

The examples in the table below were prepared from commercially available reagents using a similar method to example 4.

Example 22

N- [5- ({4- [ (2S) -2- ({ 2-cyclopropyl-7-methylthio-eno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] acetamide

Step 1: preparation of tert-butyl 4- [ (2S) -2- ({ 2-cyclopropyl-7-methylthio [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazine-1-carboxylate (intermediate 17)

To 4- [ (2S) -2-aminopropyl group]Piperazine-1-carboxylic acid tert-butyl ester hydrochloride (0.763g,2.73mmol) and 4-chloro-2-cyclopropyl-7-methylthioeno [3,2-d]A mixture of pyrimidine (0.613g,2.73mmol) in 96% EtOH (6mL) was added DIPEA (1.43mL,8.18mmol) and the mixture was heated at 85 ℃ for 35 h. The volatiles were removed under reduced pressure and the residue partitioned between DCM and water. The aqueous phase was extracted with DCM and the combined organic layers were extracted with saturated NaHCO₃Washed, dried over sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography, eluting with (DCM: MeOH ═ 99.5:0.5 to 95:5) to provide the title compound as a light yellow solid (0.870g,2.011mmol, 74% yield).

LC-MS (ESI): M/z (M +1):432.2 (method 4)

Step 2: preparation of 2-cyclopropyl-7-methyl-N- [ (2S) -1- (piperazin-1-yl) propan-2-yl ] thieno [3,2-d ] pyrimidin-4-amine (intermediate 18)

To 4- [ (2S) -2- ({ 2-cyclopropyl-7-methylthioeno [3,2-d ] cooled to 5 deg.C]Pyrimidin-4-yl } amino) propyl]To a solution of piperazine-1-carboxylic acid tert-butyl ester (intermediate 17,0.868g,2.011mmol) in DCM (15mL) was added 4M HCl in bisA solution in alkane (5.52mL,14.08mmol) and the mixture was warmed to room temperature and stirred overnight. Adding additional 4M HCl in twoA solution in alkane (0.503mL) and the mixture was stirred at room temperature for another 24 h. The volatiles were removed under reduced pressure to give 2-cyclopropyl-7-methyl-N- [ (2S) -1- (piperazin-1-yl) propan-2-yl as multiple HCl salts]Thieno [3,2-d]Pyrimidin-4-amine (0.970 g). 0.300g of this salt was dissolved in MeOH, loaded onto an SCX column (5g), and washed with MeOH. The product is treated with 2M NH₃A solution in MeOH eluted and evaporated to afford the title compound as the free base (0.200g,0.604mmol, 96% yield).

LC-MS (ESI): M/z (M +1):332.0 (method 4)

And step 3: preparation of N- [5- ({4- [ (2S) -2- ({ 2-cyclopropyl-7-methylthio-eno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] acetamide (example 22)

To the cooled to 0 ℃ 2-cyclopropyl-7-methyl-N- [ (2S) -1- (piperazin-1-yl) propan-2-yl]Thieno [3,2-d]To a solution of pyrimidin-4-amine (intermediate 18,0.070g,0.211mmol) and pyridine (0.051mL,0.634mmol) in DCM (3mL) was added 2-acetamido-1, 3-thiazole-5-sulfonyl chloride (0.051g,0.211mmol) and the mixture was warmed to room temperature and stirred for 3h. The mixture was partitioned between DCM and water. The organic phase was washed with saturated NaHCO₃Washed, dried over sodium sulfate, filtered and concentrated. The crude was purified by flash chromatography, eluting with (DCM: MeOH ═ 99.5:0.5 to 95:5) to provide the title compound as a light yellow foam (0.087g,0.162mmol, 77% yield).

LC-MS (ESI): M/z (M +1):536.1 (method 4)

¹H NMR(300MHz,DMSO-d₆)δppm 12.74(s,1H),7.95(s,1H),7.60(s,1H),7.21(d,1H),4.32-4.50(m,1H),2.92(br s,4H),2.38-2.65(m,5H),2.22-2.34(m,4H),2.20(s,3H),1.95-2.08(m,1H),1.16(d,3H),0.81-1.02(m,4H)

The examples in the table below were prepared from commercially available reagents using a method similar to that of example 22.

Example 32

N- [5- ({4- [ (2S) -2- { [ 2-chloro-7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] carbamic acid methyl ester

Step 1: preparation of tert-butyl 4- [ (2S) -2- { [ 2-chloro-7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazine-1-carboxylate (intermediate 19)

The title compound was prepared following the procedure used for the synthesis of intermediate 3, starting from 2, 4-dichloro-7- (trifluoromethyl) thieno [3,2-d ] pyrimidine [ prepared using the procedure described in WO 2013078765 compound 7-5 ] (50mg,0.183mmol) to provide the title compound (83mg,0.172mmol, 94% yield) as a white solid.

LC-MS (ESI) M/z (M +1):480.11 (method 5)

¹H NMR(400MHz,Ac-d₆)δppm 8.64(s,1H),7.42(br d,J＝7.45Hz,1H),4.55-4.73(m,1H),3.30(br t,J＝4.60Hz,4H),2.37-2.65(m,6H),1.41(s,9H),1.33(d,J＝6.58Hz,3H)

Step 2: preparation of 2-chloro-N- [ (2S) -1- (piperazin-1-yl) propan-2-yl ] -7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-amine (intermediate 20)

The title compound was prepared following the procedure used for the synthesis of intermediate 4, starting from tert-butyl 4- [ (2S) -2- { [ 2-chloro-7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazine-1-carboxylate (intermediate 19,83mg,0.17mmol) to provide the title compound as a yellowish solid (64mg,0.168mmol, 98% yield).

LC-MS (ESI) M/z (M +1):380.06 (method 5)

And step 3: preparation of methyl N- [5- ({4- [ (2S) -2- { [ 2-chloro-7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] carbamate (example 32)

To 2-chloro-N- [ (2S) -1- (piperazin-1-yl) propan-2-yl]-7- (trifluoromethyl) thieno [3,2-d]To a solution of pyrimidin-4-amine (intermediate 20,42mg,0.086mmol) in dry pyridine (0.7ml) was added N- [5- (chlorosulfonyl) -4-methyl-1, 3-thiazol-2-yl]Methyl carbamate (30mg,0.112mmol) and the solution was sonicated for 5 min. After evaporation of the pyridine, the residue was treated with heptane (35ml) and the solvent was evaporated under vacuum. The crude material was dissolved in DMF (1ml) and purified by reverse phase flash chromatography (gradient A: B from 100:0 to 0:100 with 12 column volumes, eluent A: H₂O, ACN, HCOOH 95:5:0.1, eluent B, H₂ACN HCOOH5:95: 0.1). Appropriate fractions were combined and evaporated to giveThe title compound (20.1mg,0.033mmol, 37.9% yield) as a white solid.

LC-MS (ESI) M/z (M +1):614.10 (method 5)

1H NMR (600MHz, acetone-d)₆)δppm 8.55(s,1H)7.40(br d,J＝7.87Hz,1H)4.64-4.73(m,1H)3.84(s,3H)3.09(br s,4H)2.80(br s,2H)2.73(dd,J＝12.76,8.82Hz,1H)2.59-2.68(m,2H)2.48(dd,J＝12.70,5.30Hz,1H)2.41(s,3H)1.29(d,J＝6.68Hz,3H)2.41(s,3H)1.29(d,J＝6.68Hz,3H)

Example 33

3- { [ 4-methyl-5- ({4- [ (2S) -2- ({ 7-methylthiothieno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] carbamoyl } azetidine-1-carboxylic acid tert-butyl ester

Step 1: preparation of 4-methyl-5- ({4- [ (2S) -2- ({ 7-methylthio [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-amine (intermediate 21)

Reacting N- [ 4-methyl-5- ({4- [ (2S) -2- ({ 7-methylthio-eno [3,2-d ]]Pyrimidin-4-yl } amino) propyl]Piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl]Acetamide (1.48g 2.90, mmol) dissolved in 6mL of H₂SO₄In an aqueous solution (50% w/w), the solution was stirred at 60 ℃ for 24 h. 30mL of 2N NaOH in water was added and a large amount of solid precipitated. The solid was filtered off with suction, washed with water and dried. The solid was thus recrystallized from IPA to provide the title compound (0.90g,1.92mmol, 66% yield).

LC-MS (ESI) M/z (M +1):467.98 (method 3)

Step 2: preparation of tert-butyl 3- { [ 4-methyl-5- ({4- [ (2S) -2- ({ 7-methylthio-eno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] carbamoyl } azetidine-1-carboxylate (example 34)

4-methyl-5- ({4- [ (2S) -2- ({ 7-methylthio-eno [3, 2-d)]Pyrimidin-4-yl } amino) propyl]Piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-amine (intermediate 21,87mg,0.186mmol), 1- [ (tert-butoxy) carbonyl]Azetidine-3-carboxylic acid (90mg,0.446mmol) and HATU (116mg,0.305mmol) were dissolved in DMF (1.5mL) and N-ethyl-N-isopropylpropan-2-amine (0.060mL,0.346mmol) was added in one portion. The solution was stirred at 50 ℃. The mixture was partitioned between DCM and water. The organic phase was washed with saturated NaHCO₃Washed, dried over sodium sulfate, filtered and concentrated. The crude was purified by flash chromatography on the direct phase (gradient a: B from 100:0 to 0:100 with 10 column volumes, eluent a: DCM, eluent B: DCM/MeOH ═ 90:10) to afford the title compound (71mg,0.109mmol, 65% yield) as an off-white foam.

LC-MS (ESI) M/z (M +1):651.33 (method 3)

¹H NMR (400MHz, acetone-d₆)δppm 11.37(br s,1H)8.46(s,1H)7.50(s,1H)6.40(br d,J＝7.45Hz,1H)4.61(spt,J＝6.91Hz,1H)4.02-4.21(m,4H)3.78(quin,J＝7.29Hz,1H)2.99-3.11(m,4H)2.59-2.70(m,5H)2.47(br d,J＝6.36Hz,1H)2.44(s,3H)2.33(s,3H)1.41(s,9H)1.26(d,J＝6.58Hz,3H)

Example 34

N- [5- ({4- [ (2S) -2- { [ 2-cyclopropyl-7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] carbamic acid methyl ester

Step 1: preparation of tert-butyl 4- [ (2S) -2- { [ 2-cyclopropyl-7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazine-1-carboxylate (intermediate 22)

In a 20mL microwave vial, 4- [ (2S) -2- { [ 2-chloro-7- (trifluoromethyl) thieno [3,2-d ] was loaded]Pyrimidin-4-yl]Amino group} propyl radical]Piperazine-1-carboxylic acid tert-butyl ester (100mg,0.208mmol), 2-cyclopropyl-4, 4,5, 5-tetramethyl-1, 3, 2-dioxaborolane (0.076ml,0.417mmol), and Na₂CO₃(66.2mg,0.625mmol) and suspended in 10ml of bisAlkane/water 2: 1. The mixture was heated at 100 ℃ and Pd (dppf) Cl was added₂(15.25mg,0.021mmol)。

The vial was sealed and the reaction was stirred at 110 ℃ overnight. The mixture was partitioned between AcOEt and water. The organic phase was washed with saturated NaHCO₃Washed, dried over sodium sulfate, filtered and concentrated, and the crude material is purified by flash chromatography (gradient a: B from 100:0 to 40:60 with 10 column volumes, eluent a: n-heptane, eluent B: acetone).

The appropriate fractions were combined and evaporated to give the title compound as an off-white solid (34.8mg,0.072mmol, 34.4% yield).

LC-MS (ESI) rt 0.80 min; m/z (M +1):486.18 (method 3)

Step 2-cyclopropyl-N- [ (2S) -1- (piperazin-1-yl) propan-2-yl ] -7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-amine (intermediate 23)

The title compound was prepared following the procedure used for the synthesis of intermediate 4, starting from tert-butyl 4- [ (2S) -2- { [ 2-cyclopropyl-7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazine-1-carboxylate (intermediate 22,35mg,0.17mmol) to provide the title compound as a yellowish solid (27mg,0.070mmol, 98% yield).

LC-MS (ESI) rt 0.48 min; m/z (M +1):386.13 (method 3)

And 3, step 3: preparation of methyl N- [5- ({4- [ (2S) -2- { [ 2-cyclopropyl-7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] carbamate (example 34)

To a solution of 2-cyclopropyl-N- [ (2S) -1- (piperazin-1-yl) propan-2-yl ] -7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-amine (intermediate 23,27mg,0.070mmol) in dry pyridine (0.7ml) was added methyl N- [5- (chlorosulfonyl) -4-methyl-1, 3-thiazol-2-yl ] carbamate (23mg,0.084mmol) and the solution was sonicated for 5 min. After evaporation of the pyridine, the residue was treated with heptane (35ml) and the solvent was evaporated in vacuo. The crude was purified by flash chromatography in the direct phase (gradient A: B from 100:0 to 0:100 with 10 column volumes, eluent A: DCM, eluent B: DCM: EtOH 80: 20). The appropriate fractions were combined and evaporated to give the title compound as an off-white solid (38.9mg,0.063mmol, 90% yield).

LC-MS (ESI) with rt 1.47 min; m/z (M +1):620.11 (method 5)

¹H NMR (400MHz, acetone-d₆)δppm 10.75-11.02(m,1H)8.43(s,1H)6.72(br d,J＝7.23Hz,1H)4.55-4.68(m,1H)3.85(s,3H)3.11(br t,J＝4.60Hz,4H)2.62-2.88(m,6H)2.43(s,3H)2.08-2.16(m,1H)1.29(d,J＝6.58Hz,3H)0.82-1.09(m,4H)

Example 36

N- [5- ({4- [ (2S) -2- ({ 2-ethyl-7-methylthio-eno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] acetamide

Step 1: preparation of tert-butyl 4- [ (2S) -2- ({ 2-ethyl-7-methylthio [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazine-1-carboxylate (intermediate 24)

To 4- [ (2S) -2-aminopropyl group]Piperazine-1-carboxylic acid tert-butyl ester hydrochloride (0.714g,2.55mmol) and 4-chloro-2-ethyl-7-methylthioeno [3,2-d]To a mixture of pyrimidine (0.512g,2.41mmol) in 96% EtOH (7.2mL) was added DIPEA (1.26mL) and the mixture was heated at 85 ℃ for 36 h. The volatiles were removed under reduced pressure and the residue was takenThe residue was partitioned between DCM and water. The aqueous phase was extracted with DCM and the combined organic layers were extracted with saturated NaHCO₃Washed, dried over sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography on silica gel column (DCM to DCM: MeOH ═ 95:5) to afford the title compound (yield considered quantitative).

LC-MS (ESI) M/z (M +1):420.3 (method)

Step 2-Ethyl-7-methyl-N- [ (2S) -1- (piperazin-1-yl) propan-2-yl ] thieno [3,2-d ] pyrimidin-4-amine (intermediate 25)

To the crude 4- [ (2S) -2- ({ 2-ethyl-7-methylthioeno [3,2-d ] cooled to 5 deg.C]Pyrimidin-4-yl } amino) propyl]To a solution of piperazine-1-carboxylic acid tert-butyl ester (intermediate 24, theory 2.4mmol) in DCM (16mL) was added 4M HCl in bisA solution in alkane (4mL,16.0mmol) and the mixture was warmed to room temperature and stirred overnight. Volatiles were removed under reduced pressure and the residue was dissolved in MeOH, loaded onto an SCX column (2 × 10g) and washed with MeOH. The product is treated with 2M NH₃A solution in MeOH was eluted and the volatiles were evaporated to afford the title compound as the free base (0.600g,1.88mmol, 78% yield over two steps).

LC-MS (ESI): M/z (M +1):320.1 (method 4)

And step 3: preparation of N- [5- ({4- [ (2S) -2- ({ 2-ethyl-7-methylthio-eno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] acetamide (example 36)

To the cooled to 0 ℃ 2-ethyl-7-methyl-N- [ (2S) -1- (piperazin-1-yl) propan-2-yl]Thieno [3,2-d]To a solution of pyrimidin-4-amine (intermediate 25,0.104g,0.33mmol) and pyridine (0.082mL,0.99mmol) in DCM (3mL) was added 2-acetamido-4-methyl-1, 3-thiazole-5-sulfonyl chloride (0.083g,0.33mmol) and the mixture was warmed to room temperature and stirred for 3 h. The mixture was diluted with DCM and washed with waterWater was washed and the aqueous phase was extracted with DCM. The combined organic layers were washed with saturated NaHCO₃Washed, dried over sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography eluting with DCM: MeOH 99.5:0.5 to 96:4 to provide the title compound as a white foam (0.101g,0.188mmol, 58% yield).

LC-MS (ESI) M/z (M +1) 538.2 (method 4)

¹H NMR(300MHz,DMSO-d6)δppm 12.62(s,1H),7.62(d,J＝0.98Hz,1H),7.25(d,J＝8.03Hz,1H),4.42-4.65(m,1H),2.94(br s,4H),2.70(q,J＝7.57Hz,2H),2.46-2.65(m,5H),2.45(s,3H),2.22-2.38(m,4H),2.18(s,3H),1.24(t,J＝7.57Hz,3H),1.17(d,J＝6.53Hz,3H)

The examples in the table below were prepared from commercially available reagents using a method similar to that of example 36.

Example 37

N- [5- ({4- [ (2S) -2- ({2, 7-Dimethylthio [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] carbamic acid methyl ester

Step 1: preparation of tert-butyl 4- [ (2S) -2- ({2, 7-dimethylthieno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazine-1-carboxylate (intermediate 26)

To 4- [ (2S) -2-aminopropyl group]Piperazine-1-carboxylic acid tert-butyl ester hydrochloride (1.197g,4.278mmol) and 4-chloro-2, 7-bisMethylthioeno [3,2-d]To a mixture of pyrimidine (0.850g,4.278mmol) in EtOH (9mL) was added DIPEA (2.236mL,12.835mmol) and the mixture was heated at 85 ℃ for 32 h. The mixture was cooled to room temperature, the volatiles were removed under reduced pressure, and the residue was partitioned between DCM and water. The aqueous phase was extracted with DCM and the combined organic layers were extracted with saturated NaHCO₃And brine, dried over sodium sulfate, filtered and concentrated. The residue was purified by flash chromatography on a Biotage silica gel Ultra 25g column (DCM: MeOH 99.5:0.5 to 95:5) to provide the title compound as a light yellow foam (74% yield).

LC-MS (ESI): M/z (M +1):406.0 (method 4)

Step 2, 7-dimethyl-N- [ (2S) -1- (piperazin-1-yl) propan-2-yl ] thieno [3,2-d ] pyrimidin-4-amine (intermediate 27)

To 4- [ (2S) -2- ({2, 7-dimethylthieno [3,2-d ] cooled to 5 deg.C]Pyrimidin-4-yl } amino) propyl]To a solution of piperazine-1-carboxylic acid tert-butyl ester (intermediate 26,1.5g,3.7mmol) in DCM (30mL) was added 4M HCl in bisA solution in alkane (6.5mL,25.9mmol) and the mixture was warmed to room temperature and stirred overnight. The volatiles were removed under reduced pressure to give (S) -2, 7-dimethyl-N- (1- (piperazin-1-yl) propan-2-yl) thieno [3, 2-d) as multiple HCl salts]Pyrimidin-4-amine (1.6 g). 0.300g of this salt was dissolved in MeOH, loaded onto an SCX column (5g), and washed with MeOH. The product is treated with 2M NH₃A solution in MeOH in DCM was eluted and evaporated to afford the title compound as the free base as a light yellow foam (0.220 g).

LC-MS (ESI) M/z (M +1) 306.0 (method 4)

And step 3: preparation of methyl N- [5- ({4- [ (2S) -2- ({2, 7-dimethylthieno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] carbamate (example 37)

To the cooled to 0 ℃ 2, 7-dimethyl-N- [ (2S) -1- (piperazin-1-yl) propan-2-yl group]Thieno [3,2-d]To a solution of pyrimidin-4-amine (intermediate 27,0.080g,0.262mmol) and pyridine (0.064mL,0.786mmol) in DCM (3.5mL) was added N- [5- (chlorosulfonyl) -4-methyl-1, 3-thiazol-2-yl]Methyl carbamate (0.071g,0.262mmol) and the mixture was warmed to room temperature and stirred for 3 h. The mixture was diluted with DCM and washed with water. The organic phase was washed with saturated NaHCO₃Washed, dried over sodium sulfate, filtered and concentrated. The crude was purified by flash chromatography on a silica gel Sepachrom 4g column (DCM: MeOH 99.5:0.5 to 95:5) to provide the title compound as a pink foam (0.082g,0.152mmol, 58% yield).

LC-MS (ESI): M/z (M +1):540.1 (method 4)

¹H NMR(300MHz,DMSO-d6)δppm 12.60(s,1H),7.62(s,1H),7.25(d,1H),4.41-4.57(m,1H),3.78(s,3H),2.95(br s,4H),2.46-2.67(m,5H),2.43(s,3H),2.21-2.36(m,4H),1.16(d,J＝6.50Hz,3H)

EXAMPLE 41

N- [5- ({4- [ (2S) -2- { [7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) thieno [3,2-d]Pyrimidin-4-yl]Amino } propyl group]Piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl]-N-methyl carbamic acid methyl ester

Step 1: preparation of 4-methyl-2- (methylamino) -1, 3-thiazole-5-sulfonyl chloride (intermediate 28)

To a solution of N, 4-dimethyl-1, 3-thiazol-2-amine (250mg,1.95mmol) in DCM (2.5mL) was slowly added chlorosulfonic acid (1.95mL,29.25mmol) at 25 ℃. The reaction was then stirred at 25 ℃ for 2h and then quenched with cold water and extracted with DCM (2 times). The solvent was removed under vacuum and the crude product (53mg,0.23mmol, 12% yield) was used in the next step without any further purification.

LC-MS (ESI): M/z (M +1):223 (method 1)

And 2, step: preparation of 7- (3, 5-dimethyl-1, 2-Azol-4-yl) -N- [ (2S) -1- [4- [ [ 4-methyl-2- (methylamino) -1, 3-thiazol-5-yl]Sulfonyl radical]Piperazin-1-yl]Propane-2-yl]Thieno [3,2-d]Pyrimidine-4-amine (intermediate 29)

Following the procedure used for the synthesis of example 1, starting from 7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) -N- [ (2S) -1-piperazin-1-ylpropan-2-yl]Thieno [3,2-d]The title compound was prepared starting from pyrimidin-4-amine hydrochloride (intermediate 9,70mg,0.17mmol) and 4-methyl-2- (methylamino) -1, 3-thiazole-5-sulfonyl chloride (intermediate 28,53mg,0.23mmol) to provide the title compound (36.4mg,0.06mmol, 38% yield).

LC-MS (ESI) M/z (M +1) 563.2 (method 1)

And step 3: preparation of N- [5- ({4- [ (2S) -2- { [7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) thieno [3,2-d]Pyrimidin-4-yl]Amino } propyl group]Piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl]-methyl N-methylcarbamate (example 41)

At 0 ℃ to 7- (3, 5-dimethyl-1, 2-Azol-4-yl) -N- [ (2S) -1- [4- [ [ 4-methyl-2- (methylamino) -1, 3-thiazol-5-yl]Sulfonyl radical]Piperazin-1-yl]Propane-2-yl radical]Thieno [3,2-d]Pyrimidin-4-amine (intermediate 29,36.4mg,0.06mmol) in DCM (1mL) was added N, N-dimethyl-4-pyridylamine (19.8mg,0.16mmol) followed by methyl chloroformate (0.01mL,0.130mmol) and the mixture stirred at room temperature for 2 h. The solvent was removed under reduced pressure and the crude was purified by flash chromatography eluting with 0% to 2% MeOH in DCM to give the title compound (example 41,24mg,0.039mmol, 59.7% yield).

LC-MS (ESI) M/z (M +1):621.06 (method 2)

¹H NMR(400MHz,DMSO-d6)δppm 8.41(s,1H),8.13(s,1H),7.66(d,J＝8.3Hz,1H),4.55(dt,J＝14.3,6.9Hz,1H),3.85(s,3H),3.46(s,3H),2.98(br s,4H),2.54-2.62(m,5H),2.47(s,3H),2.17-2.42(m,7H),1.17(d,J＝6.6Hz,3H)

Example 50

2-methoxy-N- [ 4-methyl-5- ({4- [ (2S) -2- { [7- (pyridin-3-yl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] acetamide

Step 1: preparation of tert-butyl 4- [ (2S) -2- [ (7-bromothieno [3,2-d ] pyrimidin-4-yl) amino ] propyl ] piperazine-1-carboxylate (intermediate 30)

The title compound was prepared following the procedure used for the synthesis of intermediate 3, starting from 7-bromo-4-chlorothieno [3,2-d ] pyrimidine (1g,4.01mmol) to provide the title compound (1.64g,3.6mmol, 90% yield).

LC-MS (ESI) M/z (M +1):458.1 (method 1)

Step 2: preparation of tert-butyl 4- [ (2S) -2- [ (7-pyridin-3-ylthieno [3,2-d ] pyrimidin-4-yl) amino ] propyl ] piperazine-1-carboxylate (intermediate 31)

To 4- [ (2S) -2- [ (7-bromothieno [3,2-d ]]Pyrimidin-4-yl) amino]Propyl radical]Piperazine-1-carboxylic acid tert-butyl ester (intermediate 30,250mg,0.55mmol), K₃PO₄To a suspension of (232mg,1.1mmol) in water (0.5mL) and THF (1.5mL) was added 3-pyridylboronic acid (101mg,0.82 mmol). After degassing with nitrogen, Pd (dppf) Cl is added₂(12mg,0.02mmol) and the tube sealed. The reaction was heated at 80 ℃ for 24 h. Water was added and the mixture was extracted with AcOEt. The organic layer was dried over sodium sulfate and concentrated under reduced pressure. The crude was purified by flash chromatography on a 28g NH column eluting with a solution from 0% to 65% AcOEt in cyclohexane to provide the title compound (158mg,0.35mmol, 64% yield).

LC-MS (ESI) M/z (M +1):455.2 (method 2)

And step 3: preparation of N- [ (2S) -1-piperazin-1-ylpropan-2-yl ] -7-pyridin-3-ylthieno [3,2-d ] pyrimidin-4-amine hydrochloride (intermediate 32)

The title compound was prepared following the procedure used for the synthesis of intermediate 9, starting from tert-butyl 4- [ (2S) -2- [ (7-pyridin-3-ylthieno [3,2-d ] pyrimidin-4-yl) amino ] propyl ] piperazine-1-carboxylate (intermediate 31,158.6mg,0.350mmol) to afford the title compound (165mg, crude) which was used in the next step without further purification.

LC-MS (ESI): M/z (M +1):355 (method 2)

And 4, step 4: preparation of N- [ 4-methyl-5- [4- [ (2S) -2- [ (7-pyridin-3-ylthieno [3,2-d ] pyrimidin-4-yl) amino ] propyl ] piperazin-1-yl ] sulfonyl-1, 3-thiazol-2-yl ] acetamide (intermediate 33)

The title compound was prepared following the procedure used for the synthesis of example 1, starting from N- [ (2S) -1-piperazin-1-ylpropan-2-yl ] -7-pyridin-3-ylthieno [3,2-d ] pyrimidin-4-amine hydrochloride (intermediate 32,136mg,0.349mmol) to provide the title compound (191.4mg,0.33mmol, 96% yield) as a white solid.

LC-MS (ESI) M/z (M +1):573.1 (method 2)

And 5: preparation of N- [ (2S) -1- [4- [ (2-amino-4-methyl-1, 3-thiazol-5-yl) sulfonyl ] piperazin-1-yl ] propan-2-yl ] -7-pyridin-3-ylthieno [3,2-d ] pyrimidin-4-amine (intermediate 34)

The compound N- [ 4-methyl-5- [4- [ (2S) -2- [ (7-pyridin-3-yl-thieno [3, 2-d)]Pyrimidin-4-yl) amino]Propyl radical]Piperazin-1-yl]Sulfonyl-1, 3-thiazol-2-yl]Acetamide (intermediate 33,191.4mg,0.33mmol) was dissolved in a mixture of sulfuric acid (1.89mL,34mmol) and water (1.9mL) and the mixture was stirred at 80 ℃ for 16 h. The mixture was cooled at 0 ℃ and water was added followed by 5M NaOH solution until pH 6. Then saturated NaHCO was added dropwise₃Solution, and a white precipitate formed. The precipitate was filtered off on a buchner funnel and washed with water to give the title compound (140mg,0.26mmol, 79% yield), which was used in the next step without further purification.

LC-MS (ESI) M/z (M +1):531.1 (method 2)

Step 6: preparation of 2-methoxy-N- [ 4-methyl-5- ({4- [ (2S) -2- { [7- (pyridin-3-yl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] acetamide (example 50)

The title compound was prepared following the procedure used for the synthesis of example 41, starting from N- [ (2S) -1- [4- [ (2-amino-4-methyl-1, 3-thiazol-5-yl) sulfonyl ] piperazin-1-yl ] propan-2-yl ] -7-pyridin-3-ylthieno [3,2-d ] pyrimidin-4-amine (intermediate 34,70mg,0.132mmol) and 2-methoxyacetyl chloride (0.01mL,0.160mmol) to provide the title compound (40mg,0.066mmol, 50% yield).

LC-MS (ESI): M/z (M +1):603.1 (method 2)

¹H NMR(400MHz,DMSO-d6)δppm 12.60(br s,1H),9.19(d,J＝1.5Hz,1H),8.56(dd,J＝4.7,1.6Hz,1H),8.51(s,1H),8.47(s,1H),8.42-8.46(m,1H),7.67(d,J＝7.9Hz,1H),7.46-7.56(m,1H),4.47-4.63(m,1H),4.15(s,2H),3.33(s,3H),2.99(br s,4H),2.54-2.63(m,5H),2.46(s,3H),2.35-2.42(m,1H),1.19(d,J＝6.6Hz,3H)

The examples in the table below were prepared from commercially available reagents using a method similar to example 50.

Example 52

N- [5- ({4- [ (2S) -2- { [7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] carbamic acid methyl ester

Step 1: preparation of N- [5- [4- [ (2S) -2- [ [7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino ] propyl ] piperazin-1-yl ] sulfonyl-1, 3-thiazol-2-yl ] acetamide (intermediate 35)

The title compound was prepared following the procedure used for the synthesis of example 1 starting from N- [ (2S) -1-piperazin-1-ylpropan-2-yl ] -7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-amine (intermediate 4,40mg,0.116mmol) to provide the title compound as a white solid (45mg,0.079mmol, 71% yield).

LC-MS (ESI) M/z (M +1) 550.2 (method 1)

Step 2: preparation of N- [ (2S) -1- [4- [ (2-amino-1, 3-thiazol-5-yl) sulfonyl ] piperazin-1-yl ] propan-2-yl ] -7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-amine (intermediate 36)

The title compound was prepared following the procedure for the synthesis of intermediate 31, starting from N- [5- [4- [ (2S) -2- [ [7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino ] propyl ] piperazin-1-yl ] sulfonyl-1, 3-thiazol-2-yl ] acetamide (intermediate 35,45mg,0.079mmol) to afford the title compound as a white solid (34mg,0.067mmol, 84% yield).

And step 3: preparation of methyl N- [5- ({4- [ (2S) -2- { [7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] carbamate (example 52)

The title compound was prepared following the procedure used for the synthesis of example 41, starting from N- [ (2S) -1- [4- [ (2-amino-1, 3-thiazol-5-yl) sulfonyl ] piperazin-1-yl ] propan-2-yl ] -7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-amine (intermediate 36,34mg,0.067mmol) and methyl chloroformate (0.01mL,0.130mmol) to provide the title compound (14mg,0.024mmol, 36% yield).

LC-MS (ESI): M/z (M +1):566 (method 2)

¹H NMR(400MHz,DMSO-d₆)δppm 12.47(s,1H),8.75(q,J＝1.1Hz,1H),8.48(s,1H),7.90(d,J＝8.1Hz,1H),7.87(s,1H),4.54(quin,J＝6.8Hz,1H),3.77(s,3H),2.93(br s,4H),2.52-2.61(m,5H),2.38(dd,J＝12.4,6.9Hz,1H),1.17(d,J＝6.4Hz,3H)

The examples in the table below were prepared from commercially available reagents using a similar method to that of example 52.

(1) Pharmacological Activity of the Compounds of the invention

In vitro assay

The effectiveness of the compounds of the invention as LPA2 antagonists on human recombinant LPA2 expressed in CHO cells can be determined using a FLIPR assay in 384-well format.

DMEM/F-12(1:1) mixture containing 2mM Glutamax supplemented with 10% fetal bovine serum, 1mM sodium pyruvate, 11mM Hepes and 1X penicillin/streptomycin in 5% CO₂The CHO-hLPA2 cell line was cultured in a humidified incubator. CHO hLPA2 cells were seeded in black-walled transparent bottom 384-well plates (#781091, Greiner Bio-One GmbH) at a density of 7,500 cells/well in 50. mu.l medium and humidity controlled CO at 37 ℃₂Overnight in an incubator. Serial dilutions of compounds were made in 100% DMSO at 200X final concentration (1:3 or 1:4,11 point CRC). Compounds were diluted 1:50 with assay buffer (20mM HEPES,145mM NaCl,5mM KCl,5.5mM glucose, 1mM MgCl2 and 2mM CaCl2, pH 7.4, containing 0.01% Pluronic F-127) before the experiment to obtain a solution (4X, 2% DMSO) corresponding to 5 times the final concentration in the assay. The final concentration of DMSO in the assay was 0.5% per well. The medium was removed by aspiration and the cells were then incubated for 30min (cell load) in an incubator at 37 ℃ with 30. mu.l of a loading solution containing 5. mu.M cytoplasmic Ca2+ indicator Cal-520 AM in assay buffer containing 2.5mM probenecid. The loaded cell plates were transferred into the FLIPR instrument and calcium responses were monitored during the online addition protocol. To test compounds, 10 μ l/well of 4X antagonist solution was added to the cells after cell loading. After 30min pre-incubation (at 37 ℃), 10 μ Ι/well of 5X concentrated LPA EC80 was added and the Ca2+ mobilization response was followed during the online addition protocol. The peak fluorescence values in the cells from which the baseline fluorescence was subtracted were output and analyzed to determine IC, respectively₅₀The value is obtained. Calcium responses were expressed as the maximum percent inhibition of EC80 agonist response.

The raw data obtained in the unstimulated control (DMSO, no LPA) was set to "100% inhibition", while the raw data obtained in the negative control (i.e., in the absence of compound and stimulated with LPA EC 80) was set to "0% inhibition".

Raw data (peak heights expressed in relative fluorescence units) were normalized and converted to "percent inhibition". Curve fitting and pIC Using XLfit software with a four parameter logistic model₅₀(-LogIC₅₀) And (6) estimating.

The results for various compounds are provided in table 2 below, wherein the compounds are classified according to their potency for inhibitory activity against LPA2 isoforms:

TABLE 2

Wherein the compounds are classified according to their potency for inhibitory activity against LPA2 isoforms according to the following classification criteria:

LPA receptor 2(LPA2)

+：LPA2 IC₅₀Less than 1000nM

++：LPA2 IC₅₀Is contained between about 100nM and 10nM

+++：LPA2 IC₅₀Less than about 10 nM.

As shown in table 2, the compounds of the present invention showed good activity as antagonists of LPA 2.

Comparative example A

(S) - (4-methyl-5- ((4- (2- ((5-methyl-7- (pyridin-3-yl) -5H-pyrrolo [3,2-d ] pyrimidin-4-yl) amino) propyl) piperazin-1-yl) sulfonyl) thiazol-2-yl) carbamic acid methyl ester

The activity of comparative example a has been tested in an in vitro assay for determining activity at LPA2 receptor as described above.

In contrast to the compounds of formula (I) according to the invention, comparative example A shows an IC of more than 1 μm, even more than 2 μm₅₀And thus the compound is inactive to the receptor LPA 2.

The above results demonstrate that the backbone (scaffold) of the compounds of formula (I) of the present invention comprising a thienopyrimidine moiety linked to piperazine through an amino-alkyl linker unexpectedly results in a series of compounds active towards the receptor LPA 2.

Claims (13)

1. A compound of formula (I)

Wherein

R is H or selected from (C)₁-C₄) Alkyl, (C)₁-C₄) Haloalkyl and 5-6 membered heteroaryl, wherein each of said heteroaryl may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, halo, (C)₁-C₄) A haloalkyl group;

R₁is H or (C)₁-C₄) An alkyl group;

R₂is H or is selected from (C)₁-C₄) Alkyl, halo, (C)₁-C₄) Haloalkyl and (C)₃-C₈) A cycloalkyl group;

R₃is H or (C)₁-C₄) An alkyl group;

a is selected from 5-6 membered heteroaryl and aryl, wherein each of said heteroaryl and aryl may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, -C (O) R₁、-C(O)OR₁、-C(O)R₁、(C₁-C₄) Haloalkyl, halo, -NR_AC(O)R₁、-NR_AC(O)OR₁、-NR_AC(O)-(C₁-C₄) alkylene-OR₁、-NR_AC(O)R_C、-NR_AC(O)NR_AR_B、-N(C₁-C₄) alkylene-NR_AR_BAryl and heteroaryl, optionally substituted by one or more (C)₁-C₄) Alkyl and (C)₁-C₄) Haloalkyl is substituted, or

When A is aryl, it may be fusedTo a second saturated or unsaturated ring optionally containing one or more heteroatoms selected from N, O and S to form a bicyclic ring system, said bicyclic ring system optionally substituted with one or more heteroatoms selected from-C (O) R₁、(C₁-C₄) Alkyl and oxo;

R_Cselected from heteroaryl, aryl, (C)₃-C₈) Cycloalkyl and (C)₄-C₈) Heterocycloalkyl, wherein said heteroaryl, aryl, heterocycloalkyl and cycloalkyl may optionally be substituted by one or more (C)₁-C₄) Alkyl and-C (O) OR₁Substitution;

R_Aand R_BIndependently at each occurrence is H or is selected from (C)₁-C₄) Alkyl, (C)₃-C₈) Cycloalkyl group, (C)₁-C₆) Haloalkyl and halo, or

R_AAnd R_BMay form, together with the nitrogen atom to which they are attached, a 4-6 membered saturated heterocyclic ring system optionally containing another heteroatom selected from N, S and O, which may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, (C)₁-C₄) A haloalkyl group and a halo group,

provided that when R is methyl and A is aryl, the aryl is not substituted with one or more methyl groups and chlorine.

2. A compound of formula (I) according to claim 1, wherein

R is H or selected from (C)₁-C₄) Alkyl, (C)₁-C₄) Haloalkyl and 5-6 membered heteroaryl, wherein each of said heteroaryl may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, halo and (C)₁-C₄) A haloalkyl group;

R₁is H or (C)₁-C₄) An alkyl group;

R₂is H or is selected from (C)₁-C₄) Alkyl, halo, (C)₁-C₄) HalogenAlkyl and (C)₃-C₈) A cycloalkyl group;

R₃is (C)₁-C₄) An alkyl group;

a is selected from 5-6 membered heteroaryl and aryl, wherein each of said heteroaryl and aryl may be optionally substituted with one or more groups selected from (C)₁-C₄) Alkyl, -C (O) R₁、-C(O)OR₁、(C₁-C₄) Haloalkyl, halo, -NR_AC(O)R₁、-NR_AC(O)OR₁、-NR_AC(O)-(C₁-C₄) alkylene-OR₁、-NR_AC(O)R_CAryl and heteroaryl, optionally substituted by one or more (C)₁-C₄) Alkyl and (C)₁-C₄) Haloalkyl is substituted, or

When a is aryl, it may be fused to a second saturated or unsaturated ring optionally containing one or more heteroatoms selected from N, O and S to form a bicyclic ring system, optionally substituted with one or more heteroatoms selected from-c (o) R₁And (C)₁-C₄) Radical substitution of alkyl;

R_Cis selected from heteroaryl and (C)₄-C₈) Heterocycloalkyl, wherein said heteroaryl and heterocycloalkyl may optionally be substituted by one or more (C)₁-C₄) Alkyl and-C (O) OR₁Substitution;

R_Ais H or (C)₁-C₄) An alkyl group, a carboxyl group,

provided that when R is methyl and A is aryl, the aryl is not substituted with one or more methyl groups and chlorine.

3. A compound of formula (I) according to claims 1 and 2, wherein a is selected from 5-6 membered heteroaryl and aryl, wherein each of said heteroaryl and aryl may be optionally substituted by one or more groups selected from (C)₁-C₄) Alkyl, -C (O) R₁、-C(O)OR₁、-C(O)R₁、(C₁-C₄) Haloalkyl, halo, -NR_AC(O)R₁、-NR_AC(O)OR₁、-NR_AC(O)-(C₁-C₄) alkylene-OR₁、-NR_AC(O)R_C、-NR_AC(O)NR_AR_B、-N(C₁-C₄) alkylene-NR_AR_BIs selected from the group consisting ofOxazole, pyridine, thiazole,Oxazole, 1,2,4-Oxadiazole, 1,3,4-Heteroaryl and aryl radicals of diazoles and pyrazoles, optionally substituted by one or more (C)₁-C₄) Alkyl and (C)₁-C₄) Haloalkyl substitution; or A is aryl which may be fused to a second saturated or unsaturated ring optionally containing one or more heteroatoms selected from N, O and S to form a bicyclic ring system optionally substituted with one or more heteroatoms selected from-C (O) R₁、(C₁-C₄) Alkyl and oxo.

4. A compound of formula (I) according to any of claims 1 to 3, wherein when a is 5-6 membered heteroaryl, said 5-6 membered heteroaryl is selected from thiazole, thiophene and furan.

5. A compound of formula (I) according to any of claims 1 to 4, wherein when R_CWhen it is heteroaryl, said heteroaryl is optionally substituted by one or more (C)₁-C₄) Alkyl and-C (O) OR₁Substituted heteroAnd (3) azole.

6. A compound of formula (I) according to claims 1-5, selected from at least one of the following:

methyl N- [ 4-methyl-5- ({4- [ (2S) -2- { [7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] carbamate,

n- [5- ({4- [ (2S) -2- { [7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) thieno [3,2-d]Pyrimidin-4-yl]Amino } propyl group]Piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl]The amino-formic acid methyl ester is obtained,

2-cyclopropyl-7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) -N- [ (2S) -1- (4- { [5- (3-methyl-1, 2-Azol-5-yl) thiophen-2-yl]Sulfonyl-piperazin-1-yl) -propan-2-yl]Thieno [3,2-d]A pyrimidine-4-amine, a salt thereof,

5- ({4- [ (2S) -2- ({ 7-methylthiophen-o [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) thiophene-2-carboxylic acid methyl ester,

7-methyl-N- [ (2S) -1- {4- [4- (2-methyl-1, 3-thiazol-4-yl) benzenesulfonyl ] piperazin-1-yl } propan-2-yl ] thieno [3,2-d ] pyrimidin-4-amine,

n- [ 5-fluoro-2-methyl-4- ({4- [ (2S) -2- ({ 7-methylthio [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) phenyl ] acetamide,

n- [ (2S) -1- [4- (3-bromobenzenesulfonyl) piperazin-1-yl ] propan-2-yl ] -7-methylthieno [3,2-d ] pyrimidin-4-amine,

n- [ (2S) -1- {4- [ (4, 5-dichlorothien-2-yl) sulfonyl ] piperazin-1-yl } propan-2-yl ] -7-methylthieno [3,2-d ] pyrimidin-4-amine,

n- [ (2S) -1- [4- (3-chloro-4-fluorobenzenesulfonyl) piperazin-1-yl ] propan-2-yl ] -7-methylthio [3,2-d ] pyrimidin-4-amine,

n- [ (2S) -1- {4- [ (4-bromo-5-chlorothien-2-yl) sulfonyl ] piperazin-1-yl } propan-2-yl ] -7-methylthieno [3,2-d ] pyrimidin-4-amine,

n- [ (2S) -1- [4- (4-bromo-3-chlorobenzenesulfonyl) piperazin-1-yl ] propan-2-yl ] -7-methylthieno [3,2-d ] pyrimidin-4-amine,

n- [ (2S) -1- {4- [ 4-chloro-3- (trifluoromethyl) benzenesulfonyl ] piperazin-1-yl } propan-2-yl ] -7-methylthieno [3,2-d ] pyrimidin-4-amine,

4-bromo-5- ({4- [ (2S) -2- ({ 7-methylthieno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) thiophene-2-carboxylic acid methyl ester,

7-methyl-N- [ (2S) -1- {4- [ (2-phenyl-1, 3-thiazol-5-yl) sulfonyl ] piperazin-1-yl } propan-2-yl ] thieno [3,2-d ] pyrimidin-4-amine,

7-methyl-N- [ (2S) -1- {4- [ (2-phenylpyrimidin-5-yl) sulfonyl ] piperazin-1-yl } propan-2-yl ] thieno [3,2-d ] pyrimidin-4-amine,

n- [4- ({4- [ (2S) -2- ({ 7-methylthiophen-o [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) phenyl ] pyridine-4-carboxamide,

7-methyl-N- [ (2S) -1- {4- [4- (4-methyl-1, 3-thiazol-2-yl) benzenesulfonyl ] piperazin-1-yl } propan-2-yl ] thieno [3,2-d ] pyrimidin-4-amine,

n- [ 2-chloro-5-fluoro-4- ({4- [ (2S) -2- ({ 7-methylthio [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) phenyl ] acetamide,

n- [ 4-methyl-5- ({4- [ (2S) -2- ({ 7-methylthio-eno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] acrylamide,

methyl N- [ 4-methyl-5- ({4- [ (2S) -2- ({ 7-methylthiophen-o [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] carbamate,

7-methyl-N- [ (2S) -1- (4- { [5- (1, 2-)Azol-5-yl) thiophen-2-yl]Sulfonyl-piperazin-1-yl) -propan-2-yl]Thieno [3,2-d]A pyrimidine-4-amine, a salt thereof,

n- [5- ({4- [ (2S) -2- ({ 2-cyclopropyl-7-methylthio-eno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] acetamide,

7-methyl-N- [ (2S) -1- {4- [ (2-methyl-1, 3-benzothiazol-6-yl) sulfonyl ] piperazin-1-yl } propan-2-yl ] thieno [3,2-d ] pyrimidin-4-amine,

1- [5- ({4- [ (2S) -2- ({ 7-methylthiophen-o [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -2, 3-dihydro-1H-indol-1-yl ] ethan-1-one,

7-methyl-N- [ (2S) -1- [4- ({1- [5- (trifluoromethyl) pyridin-2-yl ] -1H-pyrazol-4-yl } sulfonyl) piperazin-1-yl ] propan-2-yl ] thieno [3,2-d ] pyrimidin-4-amine,

n- [ 4-methyl-5- ({4- [ (2S) -2- ({ 7-methylthieno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] carboxamide,

7-methyl-N- [ (2S) -1- (4- { [5- (2-methyl-1, 3-thiazol-4-yl) thiophen-2-yl ] sulfonyl } piperazin-1-yl) propan-2-yl ] thieno [3,2-d ] pyrimidin-4-amine,

n- [ (2S) -1- (4- { [5- (3, 4-dimethyl-1, 2-Azol-5-yl) thiophen-2-yl]Sulfonyl } piperazin-1-yl) propan-2-yl]-7-methylthiothieno [3,2-d]A pyrimidine-4-amine, a salt thereof,

7-methyl-N- [ (2S) -1- [4- ({5- [ 1-methyl-5- (trifluoromethyl) -1H-pyrazol-3-yl ] thiophen-2-yl } sulfonyl) piperazin-1-yl ] propan-2-yl ] thieno [3,2-d ] pyrimidin-4-amine,

7-methyl-N- [ (2S) -1- (4- { [5- (pyridin-2-yl) thiophen-2-yl ] sulfonyl } piperazin-1-yl) propan-2-yl ] thieno [3,2-d ] pyrimidin-4-amine,

methyl N- [5- ({4- [ (2S) -2- ({ 2-cyclopropyl-7-methylthio-eno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] carbamate,

methyl N- [5- ({4- [ (2S) -2- { [ 2-chloro-7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] carbamate,

3- { [ 4-methyl-5- ({4- [ (2S) -2- ({ 7-methylthieno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] carbamoyl } azetidine-1-carboxylic acid tert-butyl ester,

methyl N- [5- ({4- [ (2S) -2- { [ 2-cyclopropyl-7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] carbamate,

n- [5- ({4- [ (2S) -2- ({ 2-cyclopropyl-7-methylthio-eno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] acetamide,

n- [5- ({4- [ (2S) -2- ({ 2-ethyl-7-methylthio-eno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] acetamide,

methyl N- [5- ({4- [ (2S) -2- ({2, 7-dimethylthieno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] carbamate,

methyl N- [5- ({4- [ (2S) -2- ({ 2-ethyl-7-methylthio-eno [3,2-d ] pyrimidin-4-yl } amino) propyl ] piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl ] carbamate,

7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) -N- [ (2S) -1- (4- { [5- (3-methyl-1, 2-)Azol-5-yl) thiophen-2-yl]Sulfonyl-piperazin-1-yl) -propan-2-yl]Thieno [3,2-d](ii) a pyrimidin-4-amine,

7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) -N- [ (2S) -1- (4- { [5- (3, 4-dimethyl-1, 2-)Azol-5-yl) thiophen-2-yl]Sulfonyl } piperazin-1-yl) propan-2-yl]Thieno [3,2-d]A pyrimidine-4-amine, a salt thereof,

N-[5- ({4- [ (2S) -2- { [7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) thieno [3,2-d]Pyrimidin-4-yl]Amino } propyl group]Piperazin-1-yl } sulfonyl) -4-methyl-1, 3-thiazol-2-yl]-methyl N-methylcarbamate,

7- (2, 4-dimethyl-1, 3-thiazol-5-yl) -N- [ (2S) -1- (4- { [5- (3-methyl-1, 2-)Azol-5-yl) thiophen-2-yl]Sulfonyl } piperazin-1-yl) propan-2-yl]Thieno [3,2-d]A pyrimidine-4-amine, a salt thereof,

n- [ (2S) -1- (4- { [5- (3-methyl-1, 2-Azol-5-yl) thiophen-2-yl]Sulfonyl } piperazin-1-yl) propan-2-yl]-7- (pyridin-4-yl) thieno [3,2-d](ii) a pyrimidin-4-amine,

n- [ (2S) -1- (4- { [5- (3-methyl-1, 2-)Azol-5-yl) thiophen-2-yl]Sulfonyl } piperazin-1-yl) propan-2-yl]-7- (pyridin-3-yl) thieno [3,2-d]A pyrimidine-4-amine, a salt thereof,

n- [ (2S) -1- (4- { [5- (3-methyl-1, 2-)Azol-5-yl) thiophen-2-yl]Sulfonyl } piperazin-1-yl) propan-2-yl]-7- (1,3, 5-trimethyl-1H-pyrazol-4-yl) thieno [3,2-d]A pyrimidine-4-amine, a salt thereof,

7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) -2-ethyl-N- [ (2S) -1- (4- { [5- (3-methyl-1, 2-)Azol-5-yl) thiophen-2-yl]Sulfonyl-piperazines-1-yl) propan-2-yl]Thieno [3,2-d]A pyrimidine-4-amine, a salt thereof,

n- [ (2S) -1- (4- { [5- (3-methyl-1, 2-)Azol-5-yl) thiophen-2-yl]Sulfonyl } piperazin-1-yl) propan-2-yl]-7- [ 1-methyl-3- (trifluoromethyl) -1H-pyrazol-4-yl]Thieno [3,2-d]A pyrimidine-4-amine, a salt thereof,

7- (3, 5-dimethyl-1, 2-)Oxazol-4-yl) -N- [ (2S) -1- (4- { [5- (3-methyl-1, 2-)Azol-5-yl) thiophen-2-yl]Sulfonyl-piperazin-1-yl) -propan-2-yl]-2- (propan-2-yl) thieno [3,2-d](ii) a pyrimidin-4-amine,

n- [5- ({4- [ (2S) -2- { [7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] acetamide,

2-methoxy-N- [ 4-methyl-5- ({4- [ (2S) -2- { [7- (pyridin-3-yl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] acetamide,

5-methyl-N- [ 4-methyl-5- ({4- [ (2S) -2- { [7- (pyridin-3-yl) thieno [3,2-d]Pyrimidin-4-yl]Amino } propyl group]Piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl]-1,2-The oxazole-3-carboxylic acid amide is,

methyl N- [5- ({4- [ (2S) -2- { [7- (trifluoromethyl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] carbamate,

methyl N- [ 4-methyl-5- ({4- [ (2S) -2- { [7- (pyridin-3-yl) thieno [3,2-d ] pyrimidin-4-yl ] amino } propyl ] piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl ] carbamate,

n- [ (2S) -1- [4- (3, 4-dichlorobenzenesulfonyl) piperazin-1-yl ] propan-2-yl ] -7- (pyridin-3-yl) thieno [3,2-d ] pyrimidin-4-amine,

5-methyl-N- [ 4-methyl-5- ({4- [ (2S) -2- { [7- (trifluoromethyl) thieno [3,2-d]Pyrimidin-4-yl]Amino } propyl group]Piperazin-1-yl } sulfonyl) -1, 3-thiazol-2-yl]-1,2-Oxazole-3-carboxamide.

7. A pharmaceutical composition comprising a compound of formula (I) according to any one of claims 1-6 in admixture with one or more pharmaceutically acceptable carriers or excipients.

8. The pharmaceutical composition of claim 7 for oral administration.

9. A compound of formula (I) according to any one of claims 1 to 6 or a pharmaceutical composition according to claims 7 and 8 for use as a medicament.

10. A compound of formula (I) or a pharmaceutical composition for use according to claim 9 in the treatment of a disease, disorder or condition associated with dysregulation of lysophosphatidic acid receptor 2(LPA 2).

11. A compound of formula (I) or a pharmaceutical composition for use according to claims 9 and 10 in the prevention and/or treatment of fibrosis and/or a disease, disorder or condition involving fibrosis.

12. A compound of formula (I) or a pharmaceutical composition for use according to claim 11 in the prevention and/or treatment of fibrosis, including pulmonary fibrosis, Idiopathic Pulmonary Fibrosis (IPF), liver fibrosis, kidney fibrosis, eye fibrosis, cardiac fibrosis, arterial fibrosis and systemic sclerosis.

13. A compound or pharmaceutical composition of formula (I) for use according to claim 12 in the prevention and/or treatment of Idiopathic Pulmonary Fibrosis (IPF).