EA044992B1 - CHEMICAL COMPOUNDS - Google Patents

Description

The present invention relates to specific new compounds and directly related prodrugs or pharmaceutically acceptable salts thereof that have inflammasome inhibitory activity and accordingly can be used in methods of treating a human or animal body. The present invention also relates to methods for preparing these compounds, to pharmaceutical compositions containing them, and to their use for the treatment of disorders in which inflammasome activity is involved, such as autoinflammatory and autoimmune diseases.

BACKGROUND OF THE INVENTION

Autoimmune diseases are associated with the overproduction of pro-inflammatory factors. One of these is interleukin-1 (IL-1), produced by activated macrophages, monocytes, fibroblasts and other components of the innate immune system such as dendritic cells. It is involved in a variety of cellular activities, including cell proliferation, differentiation and apoptosis (Seth L. al. Rev. Immunol. 2009. 27:621-68).

IL-1 family cytokines are highly active and, as important mediators of inflammation, are primarily associated with acute and chronic inflammation (Sims J. et al. Nature Reviews Immunology 10, 89102 (February 2010)). Overproduction of IL-1 is thought to be a mediator of some autoimmune and autoinflammatory diseases. Autoinflammatory diseases are characterized by recurrent and unprovoked inflammation in the absence of autoantibodies, infection, or antigen-specific T cells.

Proinflammatory cytokines of the IL-1 superfamily include IL-1a, IL-1e, IL-18 and IL-36a, β, λ and are produced in response to pathogens and other cellular stressors as part of the host innate immune response. Unlike many other secreted cytokines, which are processed and released by the standard cellular secretory apparatus consisting of the endoplasmic reticulum and Golgi apparatus, members of the IL-1 family lack the leader sequences required for entry into the endoplasmic reticulum and are therefore retained intracellularly after translation. In addition, IL-1e, IL-18 and IL-36a, β, λ are synthesized as procytokines that require proteolytic activation to become optimal ligands for binding to their cognate receptors on target cells.

In the case of IL-1a, ΓΕ-1β and IL-18, it is now understood that a multimeric protein complex known as the inflammatory site is responsible for the activation of the IL-1e and IL-18 profiles and for the extracellular release of these cytokines. The inflammasome complex typically consists of a sensor molecule such as an NLR (nucleotide oligomerizing domain (NOD)-like receptor), an adapter molecule ASC (a poptosis-associated speck-like regulatory protein containing a CARD (Capsase Recruitment Domain)), and procaspase-1. In response to various “danger signals,” including pathogen-associated patterns (PAMs) and danger-associated patterns (DAMPs), inflammatory oligomerization subunits form an intracellular supermolecular structure. PAMPs include molecules such as peptidoglycan, viral DNA or RNA, and bacterial DNA or RNA. DAMPs, on the other hand, consist of a wide range of endogenous sterile triggers, including sodium urate crystals, silica gel, alum, asbestos, fatty acids, ceramides, cholesterol crystals, and beta-amyloid peptide aggregates. Assembly of the inflammasome platform facilitates the autocatalysis of procaspase-1 to produce a highly active cysteine protease responsible for the activation and release of pro-IL-1 β and pro-IL-18. Thus, the release of these highly inflammatory cytokines is achieved only in response to the detection and response of inflammatory sensors to specific molecular danger signals.

In humans, the 22 NLR proteins are classified into four NLR subfamilies according to their N-terminal domains. NLRA contains a CARD-AT domain, NLRB (NAIP) contains a BIR domain, NLRC (including NOD1 and NOD2) contains a CARD domain, and NLRP contains a pyrin domain. Multiple members of the NLR family are associated with inflammasome formation, including NLRP1, NLRP3, NLRP6, NLRP7, NLRP12, and NLRC4 (IPAF).

Two other structurally distinct inflammasome structures containing a PYHIN domain (pyrin and HIN domain-containing protein), namely Absent In Melanoma 2 (AIM2) and IFNλ-inducible protein 16 (IFI16) (Latz et al., Nat Rev Immunol 2013 13 (6) 397-311) serve as sensors of intracellular DNA.

The requirement for assembly of the inflammasome platform to achieve activation and release of IL-1 and IL-18 from monocytes and macrophages ensures their neat organization through a 2-step process. First, the cell must encounter a priming ligand (such as the LPS ligand of the TLR4 receptor or an inflammatory cytokine such as TNFa), which leads to NFkB-dependent transcription of NLRP3, pro-IL-1e, and pro-IL-18. Newly translated procytokines remain intracellular and inactive unless the producing cells encounter a second signal leading to inflammasome activation and procaspase-1 maturation.

In addition to the proteolytic activation of pro-IL-1e and pro-IL-18, active caspase-1 also triggers a form of inflammatory cell death known as pyroptosis through the cleavage of gasdermin D. Pyroptosis allows the externalization of mature forms of IL-1 and IL-18 along with releasing molecules

- 1,044,992 alarmins (compounds that stimulate inflammation and activate innate and adaptive immunity), such as high mobility group 1 protein (HMGB1), IL-33 and IL-1a.

Although inflammasome activation appears to have evolved as an important component of host immunity to pathogens, the NLRP3 inflammasome is unique in its ability to be activated in response to endogenous sterile danger signals. Many such sterile signals have been elucidated, and their formation is associated with specific disease states. For example, uric acid crystals found in patients with gout are effective triggers for NLRP3 activation. Likewise, cholesterol crystals found in patients with atherosclerosis may also promote NLRP3 activation. Recognition of the role of sterile danger signals as activators of NLRP3 has led to the implication of IL-1 and IL-18 in various pathophysiological features, including metabolic, physiological, inflammatory, hematological and immunological disorders.

The link to human disease is best illustrated by the finding that mutations in the NLRP3 gene, which result in gain-of-function mutations, mediate a range of autoinflammatory conditions known collectively as cryopyrin-associated periodic syndromes (CAPS), including familial cold autoinflammatory syndrome ( FCAS), Muckle-Wells syndrome (MWS), and neonatal multisystem inflammatory disease (NOMID) (Hoffman et al., Nat Genet. 29(3) (2001) 301-305). Similarly, sterile transmitter-induced activation of NLRP3 has been implicated in a wide range of disorders, including joint degeneration (gout, rheumatoid arthritis, osteoarthritis), cardiometabolic (type 2 diabetes, atherosclerosis, hypertension), central nervous system (Alzheimer's disease, Parkinson's disease, multiple sclerosis ), gastrointestinal (Crohn's disease), lungs (non-alcoholic fatty liver disease, non-alcoholic hepatosteatosis, idiopathic pulmonary fibrosis).

Current treatment options for diseases implicated by IL-1 include the IL-1 receptor antagonist anakinra, the IL-1 receptor type 1 Fc-containing soluble fusion construct, the IL-1 receptor accessory protein rilonacept, and the anti-IL-1β monoclonal antibody canakinumab. . For example, canakinumab is licensed for CAPS, tumor necrosis factor receptor-associated periodic syndrome (TRAPS), hyperimmunoglobulin D syndrome (HIDS)/mevalonate kinase deficiency (MKD), familial Mediterranean fever (FMF), and gout.

Several small molecules have been reported to inhibit NLRP3 inflammasome function. For example, glyburide is a specific inhibitor of NLRP3 activation, albeit at micromolar concentrations that are unlikely to be achievable in vivo. Non-specific agents such as parthenolide, Bay 11-7082 and 3,4-methylenedioxy-b-nitrostyrene have been reported to impair NLRP3 activation but are expected to have limited therapeutic utility due to their common structural feature of being an olefin , activated by substitution with an electron-withdrawing group; which can lead to the undesirable formation of covalent adducts with protein groups containing thiols. A number of natural products, such as β-hydroxybutyrate, sulforaphane, quercetin, and salvianolic acid, have also been reported to inhibit NLRP3 activation. Similarly, numerous effectors/modulators of other molecular targets have been reported to impair NLRP3 activation, including the G protein-coupled receptor TGR5 agonists, the sodium-glucose cotransport inhibitor epigliflozin, the dopamine receptor antagonist A-68930, the serotonin reuptake inhibitor fluoxetine, the non-steroidal anti-inflammatory drugs drugs fenamate and β-adrenergic receptor blocker nebivolol. The feasibility of these molecules as therapeutic agents for the chronic treatment of NLRP3-dependent inflammatory diseases remains to be established. A number of sulfonylurea-containing molecules have previously been identified as potent and selective inhibitors of pro-IL-1e post-translational processing (Perregaux et al., J Pharmacol. Exp. Ther. 299, 187-197, 2001). The example molecule CP-456 773 from this work was recently characterized as a specific inhibitor of NLRP3 activation (Coll et al., Nat Med 21.3 (2015): 248-255.).

The invention arose from the need to provide additional compounds to specifically modulate NLRP3-dependent cellular processes. In particular, compounds with physicochemical, pharmacological and pharmaceutical properties improved over existing compounds are desirable.

The essence of the invention

According to a first aspect, the present invention relates to a compound of formula (I), or a pharmaceutically acceptable salt thereof:

Where

R1 is selected from an unsubstituted hexahydroindacene ring:

- 2 044992

where # denotes a bond to a nitrogen atom of formula (I);

R2 represents H,

R ₃ is a C _1-4 alkyl-R ₇ wherein R 7 is selected from a 5 or 6 membered monocyclic aryl ring system containing no heteroatom or containing 1, 2 or 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, and the ring system R7 is unsubstituted or substituted with 1 substituent selected from C _1-6 alkyl, C1. ₆ alkylhydroxy, OH, COCH ₃ , amino, cyano, and where R8 is a 5 or 6 membered monocyclic heteroaryl ring containing 1 or 2 heteroatoms independently selected from oxygen, nitrogen and sulfur, wherein said ring is unsubstituted or substituted with C _{1- 6} alkyl or oxo; And

R4 represents H, a C _1-6 alkyl or a C _3-6 monocyclic alkyl group.

Applicants have discovered that the compounds of the present invention are potent inhibitors of the NLRP3 inflammasome and are expected to be useful in the treatment of diseases in which inflammasome activity is involved.

In some embodiments, R ₃ is methyl or ethyl R ₇ .

Preferably, R7 is monocyclic aryl.

Preferably, R ₇ is a monocyclic aryl with a cyano substituent.

Preferably, R ₇ is a 5 or 6 membered monocyclic aryl ring containing 1, 2 or 3 heteroatoms independently selected from oxygen, nitrogen and sulfur.

In some embodiments, R ₄ is methyl or ethyl.

In accordance with another aspect of the present invention, there is provided a pharmaceutical composition comprising a compound as defined herein, or a pharmaceutically acceptable salt thereof, in admixture with a pharmaceutically acceptable diluent or carrier.

In accordance with another aspect of the present invention, there is provided a method of inhibiting the activity of the NLRP3 inflammasome in vitro or in vivo, wherein the method comprises contacting a cell with an effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein.

In accordance with a further aspect of the present invention, there is provided a method of treating a disease or disorder in which a patient in need of such treatment exhibits inflammasome activity, wherein said method comprises administering to said patient a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, specified herein, or a pharmaceutical composition as defined herein.

According to a further aspect of the present invention, there is provided a method of treating an autoinflammatory disorder, an autoimmune disorder, a neurodegenerative disease or a malignancy in a patient in need of such treatment, wherein said method comprises administering to said patient a therapeutically effective amount of a compound of formula (I), a pharmaceutically acceptable salt thereof, as defined herein, or a pharmaceutical composition as defined herein.

In accordance with another aspect of the present invention, there is provided a method of treating an autoinflammatory disorder and/or an autoimmune disorder selected from cryopyrin-associated autoinflammatory syndromes (CAPS), including familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), chronic infantile neurological cutaneous articular syndrome (CINCA), neonatal multisystem inflammatory disease (NOMID), familial Mediterranean fever and non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), gout, rheumatoid arthritis, osteoarthritis, Crohn's disease, COPD, fibrosis, obesity, diabetes 2 type, multiple sclerosis and neuroinflammation arising from protein misfolding diseases, such as prion diseases, in a patient in need of such treatment, wherein said method comprises administering to said patient a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined herein, or a pharmaceutical composition as defined herein.

According to another aspect of the present invention, there is provided a method of treating a neurodegenerative disease, such as Parkinson's disease or Alzheimer's disease, in a patient in need of such treatment, wherein said method comprises administering to said patient a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof, as defined herein, or a pharmaceutical composition as defined herein

- 3 044992 document.

In one embodiment, the composition is for use in the treatment of cancer. In especially preferred embodiments, the malignancy is selected from metastatic cancer, gastrointestinal cancer, skin cancer, non-small cell lung cancer, and colorectal adenocarcinoma.

According to a further aspect of the present invention, there is provided a compound of formula (I) or a pharmaceutically acceptable salt or pharmaceutical composition thereof as defined herein for use in the treatment of an autoinflammatory disorder, an autoimmune disorder, a neurodegenerative disease or a malignancy. In a specific embodiment, the autoinflammatory or autoimmune disorder is a cryopyrin-associated autoinflammatory syndrome (CAPS), such as, for example, familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), chronic infantile neurological cutaneous articular syndrome (CINCA) , neonatal multisystem inflammatory disease (NOMID), familial Mediterranean fever and non-alcoholic fatty liver disease (NAFLD), gout, rheumatoid arthritis, Crohn's disease, COPD, fibrosis, obesity, type 2 diabetes, multiple sclerosis, or neuroinflammation arising from diseases associated with protein misfolding, such as prion diseases. In a further embodiment, the neurodegenerative disease is Parkinson's disease or Alzheimer's disease, NASH and osteoarthritis.

In accordance with a further aspect of the present invention there is provided the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof as defined herein in the manufacture of a medicament for the treatment of an autoinflammatory disorder, an autoimmune disorder, a neurodegenerative disease or a malignancy. Accordingly, the autoinflammatory or autoimmune disorder is cryopyrin-associated autoinflammatory syndrome (CAPS), such as, for example, familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), chronic infantile neurological cutaneous articular syndrome (CINCA), multisystem inflammatory disease neonatal age (NOMID), familial Mediterranean fever and non-alcoholic fatty liver disease (NAFLD), NASH, osteoarthritis, gout, rheumatoid arthritis, Crohn's disease, COPD, fibrosis, obesity, type 2 diabetes or neuroinflammation arising from diseases associated with malfolding proteins such as prion diseases. Accordingly, the neurodegenerative disease is Parkinson's disease, or Alzheimer's disease, or multiple sclerosis.

Unless otherwise specified, all technical and scientific terms used in this description have the same meaning as commonly understood by one skilled in the art to which the present invention relates. Although methods and materials similar or equivalent to those described herein may be used in the practice or testing of the present invention, suitable methods and materials are described below. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. In the event of any conflict, this description, including definitions, will control. In addition, the substances, methods and examples are illustrative only and are not intended to be limiting.

Other features and advantages of the invention will be apparent from the following detailed description and claims.

Detailed description

The link to human disease is best illustrated by the finding that mutations in the NLRP3 gene, which result in gain-of-function mutations, mediate a range of autoinflammatory conditions known collectively as cryopyrin-associated periodic syndromes (CAPS), including familial cold autoinflammatory syndrome ( FCAS), Muckle-Wells syndrome (MWS), and neonatal multisystem inflammatory disease (NOMID) (Hoffman et al., Nat Genet. 29(3) (2001) 301-305). Similarly, sterile transmitter-induced activation of NLRP3 has been implicated in a wide range of disorders, including joint degeneration (gout, rheumatoid arthritis, osteoarthritis), cardiometabolic (type 2 diabetes, atherosclerosis, hypertension), central nervous system (Alzheimer's disease, Parkinson's disease, multiple sclerosis ), gastrointestinal (Crohn's disease), lungs (non-alcoholic fatty liver disease, non-alcoholic hepatosteatosis, idiopathic pulmonary fibrosis).

Definitions.

Unless otherwise specified, terms used in the specification and claims have the following meanings set forth below.

It should be understood that references to treatment or healing include alleviation of established symptoms of a condition. Treating or curing a status, disorder or condition thus includes: (1) preventing or delaying the onset of clinical symptoms of the status, disorder

- 4 044992 or a condition developing in a person who may have or is predisposed to a status, disorder or condition, but is not yet experiencing or exhibiting clinical or subclinical symptoms of the status, disorder or condition, (2) inhibition of the status, disorder or condition, that is, stopping, reducing or delaying the progression of the disease or its relapse (in the case of maintenance treatment) or at least one clinical or subclinical symptom thereof, or (3) reducing or attenuating the disease, that is, causing regression of a status, disorder or condition, or, at least one of its clinical or subclinical symptoms.

A therapeutically effective amount means an amount of a compound that, when administered to a mammal to treat a disease, is sufficient to effect such treatment of the disease. The therapeutically effective amount will vary depending on the compound, the disease and its severity, as well as age, weight, etc. mammal being treated.

As used herein, the term alkyl includes both straight and branched chain alkyl groups such as propyl, isopropyl and tert-butyl. However, references to individual alkyl groups, such as propyl, are specific to the straight chain embodiment only, and references to individual branched chain alkyl groups, such as isopropyl, are specific to the branched chain embodiment only. For example, _{C1_6} alkyl includes _{C1_4} alkyl, _{C1_3} alkyl, propyl, isopropyl and tert-butyl.

An alkylene, alkenylene or alkynylene group is an alkyl, alkenyl or alkynyl group that is located between and serves to connect two other chemical groups. Thus, C1_ ₆ alkylene means a linear saturated divalent hydrocarbon radical containing from one to six carbon atoms, or a branched saturated divalent hydrocarbon containing from three to six carbon atoms, for example, methylene, ethylene, propylene, 2-methylpropylene, pentylene, etc. similar.

C _2-6 alkenylene means a linear divalent hydrocarbon radical containing from two to six carbon atoms, or a branched divalent hydrocarbon radical containing from three to six carbon atoms containing at least one double bond, for example, as in ethenylene, 2,4 -pentadienylene and the like.

C ₂ _ ₆ alkynylene means a linear divalent hydrocarbon radical containing from two to six carbon atoms, or a branched divalent hydrocarbon radical containing from three to six carbon atoms containing at least one triple bond, for example, as in ethynylene, propynylene and butynylene etc.

C _3-8 cycloalkyl means a hydrocarbon ring containing from 3 to 8 carbon atoms, for example cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl or bicycloheptyl.

The term halogen refers to fluorine, chlorine, bromine and iodine.

Suitable meanings for the term C _1-6 alkoxy include methoxy, ethoxy, propoxy, isopropoxy and butoxy.

Suitable meanings for the term _C1-3 alkylamino include methylamino, ethylamino, propylamino and isopropylamino.

Suitable meanings for the term di-[C1_ ₃ alkyl]amino include dimethylamino, diethylamino, N-ethyl-N-methylamino and diisopropylamino.

The term aryl means a cyclic or polycyclic aromatic ring having from 5 to 12 carbon atoms. The term aryl includes both monovalent species and divalent species. Examples of aryl groups include, but are not limited to, phenyl, biphenyl, naphthyl and the like. For clarity, aryl is phenyl.

The term 5-membered monocyclic heteroaryl ring system is used to define a ring system in which the ring system optionally contains 1, 2 or 3 heteroatoms independently selected from oxygen, nitrogen and sulfur. Suitable examples include furyl, thiophenyl, pyrrolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, imidazolyl, oxadiazolyl, thiadiazolyl, triazolyl and tetrazolyl.

The phrase compound of the invention means the compounds described herein, both generally and specifically.

Compounds of the invention.

For the avoidance of doubt, it should be understood that where a group is qualified herein as defined above or as defined above, the said group covers the first and broadest definition found as well as any and all specific definitions for that group.

Specific compounds of the invention include, for example, compounds of formula (I) or pharmaceutically acceptable salts thereof, wherein, unless otherwise indicated, each of R1, _R2 , _R3 , R4 and any related substituent groups has any meaning as defined above.

The various functional groups and substituents constituting the compounds of formula (I) are generally selected such that the molecular weight of the compound does not exceed 1000 daltons. Most commonly, the molecular weight of the compound is less than 900, such as less than 800, or less than 750, or less

- 5 044992

700, or less than 650 daltons. More conveniently, the molecular weight is less than 600 and, for example, is 550 daltons or less.

A suitable pharmaceutically acceptable salt of the compound of the present invention is, for example, an acid addition salt of a compound of the present invention that is sufficiently basic, for example, an acid addition salt with, for example, an inorganic or organic acid, for example, hydrochloric, hydrobromic, sulfuric, phosphoric , trifluoroacetic, formic, citric, methanesulfonate or maleic acid. Further, a suitable pharmaceutically acceptable salt of the compound of the present invention that is sufficiently acidic is an alkali metal salt, such as a sodium or potassium salt, an alkaline earth metal salt, such as a calcium or magnesium salt, an ammonium salt, or a salt with an organic base, which produces a pharmaceutically acceptable a cation, for example a salt with methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris-(2-hydroxyethyl)amine.

It will be understood that the compounds of formula (I) and any pharmaceutically acceptable salts thereof include stereoisomers, mixtures of stereoisomers, polymorphs of all isomeric forms of these compounds.

Compounds that have the same molecular formula but differ in the nature or sequence of addition of their atoms or the arrangement of these atoms in space are called isomers. Isomers that differ in the arrangement of their atoms in space are called stereoisomers. Stereoisomers that are not mirror images of each other are called diastereomers, and those that are not mirror images when superimposed are called enantiomers. If a compound has an asymmetric center, for example it is bound to four different groups, then a pair of enantiomers is possible. An enantiomer can be characterized by the absolute configuration of an asymmetric center and is described by the R- and S-sequence rules of Cahn and Prelog or by the method in which the molecule rotates the plane of polarized light, designated as right-handed rotation or left-handed rotation (that is, as (+) or (-) isomers, respectively). The chiral compound may be present as a single enantiomer or as a mixture of enantiomers. A mixture containing equal proportions of enantiomers is called a racemic mixture.

The compounds of the present invention may have one or more asymmetric centers; therefore, such compounds can be obtained as individual (R)- or (S)-stereoisomers or as mixtures thereof. Unless otherwise indicated, the description or name of a particular compound in the specification and claims includes both individual enantiomers and mixtures thereof, racemic or otherwise. Methods for determining stereochemistry and separating stereoisomers are well known in the art (see discussion in Chapter 4 of Advanced Organic Chemistry, 4th edition J. March, John Wiley and Sons, New York, 2001), for example, by synthesis from optically active starting materials or by separation of the racemic form. Some of the compounds of the invention may have geometric isomeric centers (E- and Z-isomers). It should be understood that the present invention covers all optical, diastereoisomeric and geometric isomers and mixtures thereof that have inflammasome inhibitory activity.

The present invention also covers compounds of the present invention, as defined herein, that contain one or more isotopic substitutions.

It should also be understood that some compounds of formula (I) may exhibit polymorphism, and that the invention covers all such forms or mixtures thereof that have inflammasome inhibitory activity. It is well known that crystalline substances can be analyzed using traditional methods such as X-ray powder diffractometry, differential scanning calorimetry, thermogravimetric analysis, Fourier transform infrared spectroscopy (DRIFT), near-infrared spectroscopy (NIR), solution nuclear magnetic resonance spectroscopy and/or solid. The water content of such crystalline substances can be determined using Karl Fischer analysis.

The compounds of formula (I) may exist in various tautomeric forms, and references to compounds of formula I include all such forms. For the avoidance of doubt, when a compound may exist as one of several tautomeric forms and only one is specifically described or shown, all others are nevertheless covered by formula (I). Examples of tautomeric forms include keto, enol, and enolate forms, as in the following tautomeric pairs: keto/enol (shown below), imine/enamine, amide/imino alcohol, amidine/amidine, nitroso/oxime, thioketone/ethiol, and nitro/aci-nitro.

,O \ OH n ⁺ h O'-s-s'c=c' - c=c ^z

I ^{4 x 4} n ⁺ / \ keto enol enolate

Compounds of formula (I) containing an amino function can also form Noxides. Reference herein to a compound of formula I which contains an amino group also includes an N-oxide. When a compound contains several amino functional groups, one or more nitrogen atoms can be oxidized to form an N-oxide. Specific examples of Noxides are N-oxides of a tertiary amine or the nitrogen atom of a nitrogen-containing heterocycle. N-oxides can be prepared by treating the corresponding amine with an oxidizing agent such as hydrogen peroxide or a peracid (eg, peroxycarboxylic acid), see, for example, Advanced Organic Chemistry, Jerry March, 4th Edition, Wiley Interscience, pages. More specifically, N-oxides can be prepared by the procedure of L. W. Deady (Syn. Comm. 1977, 7, 509-514), in which the amine compound is reacted with mchloroperoxybenzoic acid (mCPBA), for example, in an inert solvent such as dichloromethane.

The compounds of formula (I) can be administered in the form of a prodrug, which is cleaved in the human or animal body to release the compound of the present invention. A prodrug can be used to alter the physical properties and/or pharmacokinetic properties of a compound of the present invention. A prodrug can be formed when a compound of the present invention contains a suitable group or substituent to which a property-modifying group can be attached. Examples of prodrugs include in vivo cleavable ester derivatives, which can be formed at the carboxy group or hydroxy group in the compound of formula (I), and in vivo cleavable amide derivatives, which can be formed at the carboxy group or amino group in the compound of formula (I).

Accordingly, the present invention includes those compounds of formula (I) as defined above when they are made available by organic synthesis and when they are made available in the human or animal body by cleavage of their prodrugs. Accordingly, the present invention includes those compounds of formula (I) that are produced by organic synthetic means, as well as those compounds that are produced in the human or animal body by metabolism of a precursor compound, that is, the compound of formula (I) may be a synthetically produced compound or metabolically produced compound.

A suitable pharmaceutically acceptable prodrug of a compound of formula (I) is one that is based on reasonable medical judgment as being suitable for administration to a human or animal body without undesirable pharmacological activity and without undue toxicity. Various forms of the prodrug have been described, for example, in the following documents:-

a) Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et al. (Academic Press, 1985);

b) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985);

c) A Textbook of Drug Design and Development, edited by Krogsgaard-Larsen and H. Bundgaard, Chapter 5 Design and Application of Prodrugs, by H. Bundgaard p. 113-191 (1991);

d) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992);

e) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285 (1988);

f) N. Kakeya, et al., Chem. Pharm. Bull. 32, 692 (1984);

g) T. Higuchi and V. Stella, ProDrugs as Novel Delivery Systems, A.C.S. Symposium Series, Volume 14; and

h) E. Roche (editor), Bioreversible Carriers in Drug Design, Pergamon Press, 1987.

A suitable pharmaceutically acceptable prodrug of a compound of formula (I) which has a carboxy group is, for example, its in vivo cleavable ester. The in vivo cleavable ester of a compound of formula (I) containing a carboxy group is, for example, a pharmaceutically acceptable ester that is cleaved in the human or animal body to form the parent acid. Suitable pharmaceutically acceptable esters at the carboxy group include _{C1_6} alkyl esters such as methyl, ethyl and t-butyl, _C1 . ₆ α-alkoxymethyl esters, such as methoxymethyl esters, C ₁ . ₆ alkanoyloxymethyl esters such as pivaloyloxymethyl esters, 3-phthalidyl esters, C ₃ . ₈ cycloalkylcarbonyloxy-C ₁ . ₆ alkyl esters such as cyclopentylcarbonyloxymethyl and 1-cyclohexylcarbonyloxyethyl esters, 2-oxo1,3-dioxolenylmethyl esters such as 5-methyl-2-oxo-1,3-dioxolen-4-ylmethyl esters and C ₁ . ₆ alkoxycarbonyloxy-C ₁ . ₆ alkyl esters such as methoxycarbonyloxymethyl and 1-methoxycarbonyloxyethyl esters.

A suitable pharmaceutically acceptable prodrug of a compound of formula (I) which has a hydroxy group is, for example, its in vivo cleavable ester or ether. An in vivo cleavable ester or ether of a compound of formula (I) containing a hydroxy group is, for example, a pharmaceutically acceptable ester or ether that is cleaved in the human or animal body to form the parent hydroxy compound. Suitable groups forming a pharmaceutically acceptable hydroxy ester include inorganic esters such as phosphate esters (including phosphoramide cyclic esters). Other suitable groups forming a pharmaceutically acceptable hydroxy ester include C1_ ₁₀ alkanoyl groups such as acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl groups, C ₁ . ₁₀ alkoxycarbonyl groups such as ethoxycarbonyl, N, N-( _C1.6 ) ₂ -carbamoyl, 2-dialkylaminoacetyl and 2 _- carboxyacetyl groups. Examples of ring substituents on the phenyl acetyl and benzoyl groups include aminomethyl, N-alkylaminomethyl, N,Ndialkylaminomethyl, morpholinomethyl, piperazin-1-ylmethyl and 4-(C _1-4 alkyl)piperazin-1-ylmethyl. Suitable groups forming a pharmaceutically acceptable hydroxy ether include α-acyloxyalkyl groups such as acetoxymethyl and pivaloyloxymethyl groups.

A suitable pharmaceutically acceptable prodrug of a compound of formula (I) which has a carboxy group is, for example, its in vivo cleavable amide, for example an amide formed with an amine such as ammonia, C ₁ . ₄ αlkylamine such as methylamine, (C ₁ . ₄ αlkyl) ₂ amine such as dimethylamine, N-ethyl-N-methylamine or diethylamine, C1. ₄ alkoxy-C ₂ . ₄ alkylamine such as 2-methoxyethylamine, phenyl-C1_ ₄ alkylamine such as benzylamine, and amino acids such as glycine or an ester thereof.

A suitable pharmaceutically acceptable prodrug of a compound of formula (I) which possesses an amino group is, for example, its in vivo cleavable amide derivative. Suitable pharmaceutically acceptable amides at the amino group include, for example, the amide formed with C ₁ . ₁ oalkanoyl groups such as acetyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl groups. Examples of ring substituents on the phenylacetyl and benzoyl groups include aminomethyl, N-alkylaminomethyl, N,N-dialkylaminomethyl, morpholinomethyl, piperazin-1-ylmethyl and 4-(C1_ ₄ alkyl)piperazin-1-ylmethyl.

The in vivo effect of a compound of formula (I) may be due in part to one or more metabolites that are formed in the human or animal body following administration of a compound of formula (I). As stated above, the in vivo effects of a compound of formula (I) may also be exerted by metabolism of the precursor (prodrug) compound.

Although the present invention may relate to any compound or specific group of compounds defined herein by optional, preferred or suitable features or otherwise with respect to specific embodiments, the present invention may also relate to any compound or specific group of compounds that specifically exclude the specified optional, preferred or suitable features or specific embodiments. The feature of the invention relates to specific structural groups on R1, which falls within the scope of the claims as defined herein. In some cases, specific groups define structures that are not relevant to the present invention and thus may be rejected. Such structures can be rejected when R1 corresponds to phenyl directly substituted by at least 2 groups, including: 1 halogen group and 1 methyl group; 2 or more halogen groups; or 2 methyl groups.

Accordingly, the present invention excludes any individual compounds that do not have biological activity as defined herein.

General methods of obtaining.

The compounds of the present invention can be prepared by any suitable method known in the art. Specific methods for preparing these compounds are described below in the accompanying examples.

In the description of synthetic methods presented herein, and in any reference synthetic methods that are used to prepare starting materials, it should be understood that all proposed reaction conditions, including solvent selection, reaction atmosphere, reaction temperature, run time, and workup methods, may be selected by a specialist in this field.

One skilled in the art of organic synthesis will appreciate that the functional groups present in the various parts of the molecule must be compatible with the reagents and reaction conditions used.

It will be understood that during the synthesis of the compounds of the present invention in the methods described herein or during the synthesis of particular starting materials, it may be desirable to protect certain substituent groups to prevent their undesirable reaction. An experienced chemist will understand when such protection is required and how such protecting groups can be created and subsequently removed. For examples of protecting groups, see one of the many general reviews on this topic, such as Protective Groups in Organic Synthesis by Theodora Green (publisher: John Wiley & Sons). Protecting groups can be removed by any convenient method described in the literature or known to the skilled chemist depending on the need for removal of the protecting group in question, such methods being selected to ensure removal of the protecting group with minimal impact on groups elsewhere in the molecule. Thus, if the reactants include, for example, groups such as amino, carboxy or hydroxy, it may be desirable to protect the group in some of the reactions mentioned herein.

For example, a suitable protecting group for an amino or alkylamino group is, for example, an acyl group such as an alkanoyl group such as acetyl, an alkoxycarbonyl group such as a methoxycarbonyl, ethoxycarbonyl or tert-butoxycarbonyl group, an arylmethoxycarbonyl group such as benzyloxycarbonyl, or an aroyl group such as , benzoyl. Conditions

- 8 044992 deprotections for the above protective groups necessarily vary depending on the choice of protecting group. Thus, for example, an acyl group such as an alkanoyl or alkoxycarbonyl group or an aroyl group can be removed, for example, by hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium or sodium hydroxide. Alternatively, an acyl group such as a tert-butoxycarbonyl group may be removed, for example by treatment with a suitable acid such as hydrochloric, sulfuric or phosphoric acid or trifluoroacetic acid, and an arylmethoxycarbonyl group such as a benzyloxycarbonyl group may be removed, for example hydrogenation over a catalyst such as palladium on carbon, or treatment with a Lewis acid such as boron tris(trifluoroacetate). A suitable alternative protecting group for the primary amino group is, for example, a phthaloyl group, which can be removed by treatment with an alkylamine, for example dimethylaminopropylamine, or hydrazine.

A suitable protecting group for the hydroxy group is, for example, an acyl group, eg an alkanoyl group such as acetyl, an aroyl group such as benzoyl, or an arylmethyl group such as benzyl. The deprotection conditions for the above protecting groups necessarily vary depending on the choice of protecting group. Thus, for example, an acyl group, such as an alkanoyl or aroyl group, can be removed, for example, by hydrolysis with a suitable base such as an alkali metal hydroxide, for example lithium hydroxide, sodium hydroxide, or ammonia. Alternatively, an arylmethyl group, such as a benzyl group, can be removed, for example, by hydrogenation over a catalyst such as palladium on carbon.

A suitable protecting group for the carboxy group is, for example, an esterifying group, for example a methyl or ethyl group, which can be removed, for example, by hydrolysis with a base such as sodium hydroxide, or, for example, a tert-butyl group, which can be removed, for example , treatment with an acid, for example an organic acid such as trifluoroacetic acid, or, for example, a benzyl group, which can be removed, for example, by hydrogenation over a catalyst such as palladium on carbon.

Once a compound of formula (I) has been synthesized using any of the methods defined herein, those methods may further include the additional steps of:

(i) removing any protecting groups present;

(ii) converting a compound of formula (I) into another compound of formula (I);

(iii) obtaining a pharmaceutically acceptable salt, hydrate or solvate; and/or (iv) obtaining a prodrug thereof.

The resulting compounds of formula (I) can be isolated and purified using methods well known in the art.

Conveniently, the reaction of the compounds is carried out in the presence of a suitable solvent, which is preferably inert under the appropriate reaction conditions. Examples of suitable solvents include, but are not limited to, hydrocarbons such as hexane, petroleum ether, benzene, toluene or xylene; chlorinated hydrocarbons such as trichlorethylene, 1,2dichloroethane, carbon tetrachloride, chloroform or dichloromethane; alcohols such as methanol, ethanol, isopropanol, n-propanol, n-butanol or tert-butanol; ethers such as diethyl ether, diisopropyl ether, tetrahydrofuran (THF), 2-methyltetrahydrofuran, cyclopentyl methyl ether (CPME), methyl tert-butyl ether (MTBE) or dioxane; glycol ethers such as ethylene glycol monomethyl or monoethyl ether or ethylene glycol dimethyl ether (diglyme); ketones such as acetone, methyl isobutyl ketone (MIBK) or butanone; amides such as acetamide, dimethylacetamide, dimethylformamide (DMF) or N-methylpyrrolidinone (NMP); nitriles such as acetonitrile; sulfoxides such as dimethyl sulfoxide (DMSO); nitro compounds such as nitromethane or nitrobenzene; esters such as ethyl acetate or methyl acetate, or mixtures of these solvents or mixtures with water.

The reaction temperature is typically from about -100°C to 300°C, depending on the reaction stage and conditions used.

Reaction times typically range from a fraction of a minute to several days, depending on the reactivity of the compounds involved and the respective reaction conditions. The appropriate reaction time is easily determined by methods known in the art, for example, by monitoring the reaction. Based on the above reaction temperatures, suitable reaction times are usually in the range of 10 minutes to 48 hours.

In addition, using the methods described herein, in combination with conventional techniques in the art, additional compounds of the present invention can be easily prepared. Those skilled in the art will readily appreciate that known variations of the conditions and methods of the following preparation methods can be used to prepare these compounds.

As will be appreciated by one skilled in the art of organic synthesis, the compounds of the present invention are readily available through a variety of synthetic routes, some of which are illustrated in the accompanying examples. One skilled in the art will readily understand which reagents and reaction conditions must be used and how they must be applied and adapted in any particular case - wherever necessary or useful - to prepare the compounds of the present invention. Besides,

- 9 044992 some of the compounds of the present invention can be easily synthesized by reacting other compounds of the present invention under suitable conditions, for example, by converting one particular functional group present in the compound of the present invention or in a suitable precursor molecule to another, using standard synthetic methods such as reduction, oxidation, addition or substitution reactions; these methods are well known to those skilled in the art. Likewise, one skilled in the art will employ - when necessary or useful - synthetic protecting (or protecting) groups; suitable protecting groups, as well as methods for their introduction and removal, are well known to one skilled in the art of chemical synthesis and are described in more detail, for example, P.G.M. Wuts, T.W. Greene, Greene's Protective Groups in Organic Synthesis, 4th edition (2006) (John Wiley & Sons).

Pharmaceutical compositions.

According to a further aspect of the invention, there is provided a pharmaceutical composition which contains a compound of the present invention as defined above, or a pharmaceutically acceptable salt, hydrate or solvate thereof, in combination with a pharmaceutically acceptable diluent or carrier.

The compositions of the present invention may be in a form suitable for oral administration (for example, tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical application (for example, as creams, ointments, gels, or aqueous or oily solutions or suspensions), for administration by inhalation (eg, as a fine powder or liquid aerosol), for administration by insufflation (eg, as a fine powder), or for parenteral administration (eg , as a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular, intraperitoneal or intramuscular administration or as a suppository for rectal administration).

The compositions of the invention can be prepared by conventional methods using conventional pharmaceutical excipients well known in the art. Thus, compositions intended for oral administration may contain, for example, one or more coloring agents, sweeteners, flavoring agents and/or preservatives.

An effective amount of a compound of the present invention for use in therapy is an amount sufficient to treat or prevent an inflammasome-related condition referred to herein, slow its progression, and/or reduce symptoms associated with the condition.

The amount of active ingredient that is combined with one or more excipients to form a unit dosage form will necessarily vary depending on the individual being treated and the particular route of administration. For example, a formulation intended for oral administration to humans will typically contain, for example, 0.5 mg to 0.5 g of active agent (more preferably, 0.5 to 100 mg, e.g., 1 to 30 mg) in combination with a suitable and convenient amount of excipients, which may vary from about 5 to about 98 percent by weight of the total composition.

The dose size for therapeutic purposes of the compounds of the invention will naturally vary depending on the nature and severity of the conditions, the age and sex of the animal or patient and the route of administration in accordance with well known principles of medicine.

When using a compound of the present invention for therapeutic or prophylactic purposes, it is typically administered so as to achieve a daily dose in the range of, for example, 0.1 mg/kg to 75 mg/kg body weight, given in divided doses if required. In general, lower doses will be used when administered parenterally. Thus, for example, for intravenous or intraperitoneal administration, a dose in the range of, for example, 0.1 mg/kg to 30 mg/kg body weight will typically be used. Likewise, for administration by inhalation, a dose in the range of, for example, 0.05 mg/kg to 25 mg/kg body weight will be used. Oral administration, especially in tablet form, may also be suitable. Typically, unit dosage forms will contain from about 0.5 mg to 0.5 g of a compound of the present invention.

Therapeutic uses and applications.

The present invention relates to compounds that act as inhibitors of inflammasome activity. The present invention therefore provides a method of inhibiting inflammasome activity in vitro or in vivo, wherein the method comprises contacting a cell with an effective amount of a compound or a pharmaceutically acceptable salt thereof, as defined herein.

The effectiveness of the compounds of the invention can be determined using conventional assay/disease models in accordance with standard practice of ascertaining the same as described in the art and found in current general knowledge.

The present invention also relates to a method of treating a disease or disorder in which the activity of an inflammasome is involved in a patient in need of such treatment, wherein said

- 10 044992 the method includes administering to a specified patient a therapeutically effective amount of a compound or a pharmaceutically acceptable salt or pharmaceutical composition thereof as defined herein.

In general, the compounds of the present invention that inhibit the maturation of IL-1 family cytokines are effective in all therapeutic indications that are mediated by or associated with increased levels of active forms of cytokines belonging to the IL-1 family. cytokines (Sims J. et al. Nature Reviews Immunology 10, 89-102 (February 2010).

The following examples of diseases and corresponding references are given: autoinflammatory and autoimmune diseases such as CAPS (Dinarello CA. Immunity. 2004 Mar;20(3):243-4; Hoffman HM. al. Reumatologia 2005; 21(3)), gout, rheumatoid arthritis (Gabay C et al. Arthritis Research & Therapy 2009, 11:230; Schett G. et al. Nat Rev Rheumatol. 2016 Jan; 12(1): 14-24.), Crohn's disease (Jung Mogg Kim Korean J Gastroenterol Vol. 58 No. 6, 300-310), COPD (Mortaz E. et al. Tanaffos. 2011; 10(2): 9-14.), fibrosis (Gasse P. et al. Am J Respir Crit Care Med . 2009 May 15;179(10):903-13), obesity, type 2 diabetes ((Dinarello CA. et al. Curr Opin Endocrinol Diabetes Obes. 2010 Aug;17(4):314-21)), multiple sclerosis (see EAE model in Coll RC. et al. Nat Med. 2015 Mar;21(3):248-55) and many others (Martinon F. et al. Immunol. 2009. 27:229-65), such such as Parkinson's disease or Alzheimer's disease (Michael T. et al. Nature 493, 674-678 (31 January 2013); Halle A. et al., Nat Immunol. 2008 Aug;9(8):857-65; Saresella M. et al. Mol Neurodegener. 2016 Mar 3;11:23) and even some cancer disorders.

Accordingly, the compounds of the present invention can be used to treat a disease selected from the group consisting of an autoinflammatory disease, an autoimmune disease, a neurodegenerative disease and a malignancy. Said autoinflammatory and autoimmune disease is suitably selected from the group consisting of NASH, osteoarthritis, malignancy, cryopyrin-associated periodic syndrome (CAPS) (such as, for example, familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), chronic infantile neurological cutaneous articular syndrome (CINCA)/neonatal multisystem inflammatory disease (NOMID), familial Mediterranean fever and non-alcoholic fatty liver disease (NAFLD), gout, rheumatoid arthritis, Crohn's disease, COPD, fibrosis, obesity, type 2 diabetes , multiple sclerosis, and neuroinflammation that occurs in protein misfolding diseases such as prion diseases. Said neurodegenerative disease is suitably selected from Parkinson's disease and Alzheimer's disease.

Accordingly, the compounds of the present invention can be used to treat a disease selected from the group consisting of cryopyrin-associated periodic syndrome (CAPS), such as, for example, familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), chronic Infantile Neurological Cutaneous Articular Syndrome (CINCA), Neonatal Multisystem Inflammatory Disease (NOMID), Familial Mediterranean Fever and Non-Alcoholic Fatty Liver Disease (NAFLD), Gout, Rheumatoid Arthritis, Crohn's Disease, COPD, Fibrosis, Obesity, Type 2 Diabetes, Multiple sclerosis, neuroinflammation that occurs in protein misfolding diseases such as prion disease, Parkinson's disease and Alzheimer's disease.

Treatment for cancer; connections with the inflammasome.

It has long been noted that chronic inflammasome reactions are associated with various types of cancer. During malignant transformation or cancer therapy, inflammasomes can be activated in response to danger signals, and this activation can be either beneficial or detrimental in malignancy.

IL-1 expression is elevated in a variety of cancers (including breast, prostate, colon, lung, head and neck, and melanoma), and patients with IL-1-producing tumors generally have a worse prognosis (Lewis, Anne M., et al., Interleukin-1 and cancer progression: the emerging role of interleukin-1 receptor antagonist as a novel therapeutic agent in cancer treatment. Journal of translational medicine 4.1 (2006): 48).

Cancers arising from epithelial cells (carcinoma) or epithelium in glands (adenocarcinoma) are heterogeneous; made up of many different types of cells. This may include fibroblasts, immune cells, adipocytes, endothelial cells, and pericytes, among others, which may be cytokine/chemokine secreters (Grivennikov, Sergei I, Florian R. Greten, and Michael Karin. Immunity, inflammation, and cancer. Cell 140, 6 (2010): 883-899). This may lead to inflammation associated with malignancy through the infiltration of immune cells. The presence of leukocytes in tumors is known, but only recently has it become apparent that the inflammatory microenvironment is an important component of all tumors.

Most tumors (>90%) are the result of somatic or environmental mutations rather than germline mutations, and many environmental causes of malignancy are associated with chronic inflammation (20% of cancers are associated with chronic infection, 30% with smoking/inhalation of pollutants and 35% with nutritional factors (20% of all types

- 11 044992 cancers are associated with obesity) (Aggarwal, Bharat V., R. V. Vijayalekshmi, and Bokyung Sung. Targeting inflammatory pathways for prevention and therapy of cancer: short-term friend, long-term foe. Clinical Cancer

Research 15.2 (2009): 425-430).

Gastrointestinal cancer.

Gastrointestinal (GI) cancers are often associated with chronic inflammation. For example, H. pylori infection is associated with gastric cancer (Amieva, Manuel, and Richard M. Peek. Pathobiology of Helicobacter pylori-Induced Gastric Cancer. Gastroenterology 150.1 (2016): 64-78). Colorectal cancer is associated with inflammatory bowel disease (Bernstein, Charles N., et al. Cancer risk in patients with inflammatory bowel disease. Cancer 91.4 (2001): 854-862). Chronic inflammation in the stomach leads to upregulation of IL-1 and other cytokines (Basso D, et al., (1996). Helicobacter pylori infection enhances mucosal interleukin-1 beta, interleukin-6, and the soluble receptor of interleukin-2. Int J Clin Lab Res 26: 207210), and polymorphisms in the IL-1 gene may increase the risk of gastric cancer (Wang P, et al., (2007) Association of interleukin-1 gene polymorphisms with gastric cancer: a meta-analysis. Int J Cancer 120:552–562).

In 19% of gastric cancers, caspase-1 expression is decreased, which correlates with stage, lymph node metastasis, and survival (Jee et al., 2005). Mycoplasma hyorhinis is associated with the development of gastric cancer, and activation of the NLRP3 inflammasome may be associated with its activation of gastric cancer metastasis (Xu et al., 2013).

Malignant neoplasms of the skin.

Ultraviolet radiation poses the greatest environmental risk for skin cancer, which is caused by DNA damage, immunosuppression and inflammation. The most malignant skin cancer, melanoma, is characterized by increased activity of inflammatory cytokines, all of which can be regulated by IL-1 (Lazar-Molnar, Eszter, et al. Autocrine and paracrine regulation by cytokines and growth factors in melanoma. Cytokine 12.6 ( 2000): 547-554). Systemic inflammation induces increased metastasis and growth of melanoma cells through IL-1-dependent mechanisms in vivo. Thymoquinone suppresses metastasis of melanoma cells by inhibition of NLRP3 inflammasome. Toxicology and applied pharmacology 270.1 (2013): 70 -76).

Glioblastoma.

NLRP3 promotes radiotherapy resistance in glioma. Ionizing radiation can induce NLRP3 expression, whereas NLRP3 inhibition reduced tumor growth and prolonged mouse survival after radiation therapy. Therefore, inhibition of the NLRP3 inflammasome may provide a therapeutic strategy for radiation-resistant glioma (Li, Lianling and Yuguang Liu). Aging-related gene signature regulated by Nlrp3 predicts glioma progression. American journal of cancer research 5.1 (2015): 442).

Metastasis.

In a broader sense, applicants believe that NLRP3 is involved in the development of metastasis, and therefore modulation of NLRP3 should actually block this. IL-1 is involved in tumor genesis, tumor invasiveness, metastasis, interactions with the tumor host (Apte, Ron N., et al. The involvement of IL-1 in tumorigenesis, tumor invasiveness, metastasis and tumor-host interactions. Cancer and Metastasis Reviews 25.3 (2006): 387-408) and angiogenesis (Voronov, Elena, et al. IL-1 is required for tumor invasiveness and angiogenesis. Proceedings of the National Academy of Sciences 100.5 (2003): 2645-2650) .

The IL-1 gene is frequently expressed in metastases from patients with several types of human cancer. For example, IL-1 mRNA was highly expressed in more than half of all human metastatic tumor samples tested, including, but not limited to, non-small cell lung cancer, colorectal adenocarcinoma, and melanoma tumor samples (Elaraj, Dina M., et al. The role of interleukin 1 in growth and metastasis of human cancer xenografts. Clinical Cancer Research 12.4 (2006): 1088-1096), and IL1RA inhibits xenograft growth in IL-1-producing tumors, but without antiproliferative effect in vitro.

In addition, IL-1 signaling is a biomarker for predicting breast cancer patients at increased risk of developing bone metastasis. In mouse models, IL-1 and its receptor are upregulated in breast cancer cells that metastasize to bone compared to cells that do not. In a mouse model, the IL-1 receptor antagonist anakinra reduced proliferation and angiogenesis in addition to having significant effects on the tumor environment by reducing markers of bone turnover, IL-1 and TNF-alpha (Holen, Ingunn et al. IL-1 drives breast cancer growth and bone metastasis in vivo.Oncotarget (2016).

IL-18 induced MMP-9 production in the human leukemia cell line HL-60, thereby promoting extracellular matrix degradation and cancer cell migration and invasiveness (Zhang, Bin, et al. IL-18 increases invasiveness of HL-60 myeloid leuk cellsemia: up-regulation of matrix metalloproteinases-9 (MMP-9) expression. Leukemia research 28.1 (2004): 91-95). In addition, IL-18 may support the development of tumor metastasis in the liver by inducing VCAM-1 expression in the sinusoidal endothelium of the liver (Carrascal, Maria Teresa, et al. Interleukin-18 binding protein reduces b16 melanoma he- 12 044992 pathic metastasis by neutralizing adhesion and growth factors of sinusoidal endothelium. Cancer Research

63.2 (2003): 491-497).

CD36.

The fatty acid scavenger receptor CD36 has a dual role in initiating pro-IL-1 gene transcription and inducing assembly of the NLRP3 inflammasome complex. CD36 and the TLR4-TLR6 heterodimer recognize oxLDL, which initiates a signaling pathway leading to transcriptional activation of NLRP3 and προ-IL-I (signal 1). CD36 also mediates the internalization of oxLDL into the lysosomal compartment, where crystals are formed that induce lysosomal rupture and NLRP3 inflammasome activation (signal 2) (Kagan, J. and Horng T., NLRP3 inflammasome activation: CD36 serves double duty. Nature immunology 14.8 ( 2013): 772-774).

A subpopulation of human oral carcinoma cells express high levels of the fatty acid scavenger receptor CD36 and have the unique ability to initiate metastasis. Palmitic acid or a high-fat diet increased the metastatic potential of CD36+ cells. Neutralizing anti-CD36 antibodies blocked metastasis in orthotopic mouse models of human oral cancer. The presence of CD36+ metastasis-initiating cells correlates with poor prognosis for many types of carcinomas. It has been suggested that dietary lipids may promote metastasis (Pasqual, G, Avgustinova, A., Mejetta, S, Martin, M, Castellanos, A, Attolini, CS-O, Berenguer, A., Prats, N, Toll, A, Hueto , JA, Bescos, C, Di Croce, L, and Benitah, S.A. 2017 Targeting metastasis-initiating cells through the fatty acid receptor CD36 Nature 541:41-45).

In hepatocellular carcinoma, exogenous palmitic acid activated an epithelial-mesenchymal transition (EMT)-like program and induced migration, which was reduced by the CD36 inhibitor, sulfo-N-succinimidyl oleate (Nath, Aritro, et al. Elevated free fatty acid uptake via CD36 promotes epithelial- mesenchymal transition in hepatocellular carcinoma. Scientific reports 5, 2015). Body mass index was not associated with degree of EMT, highlighting that CD36 and free fatty acids are actually important.

Cancer stem cells (CSCs) use CD36 to promote their maintenance. Oxidized phospholipids, ligands of CD36, were present in glioblastoma, and proliferation of CSCs, but not CSCs, increased upon exposure to oxidized LDL. CD36 also correlates with patient prognosis.

Chemotherapy resistance.

In addition to direct cytotoxic effects, chemotherapeutic agents exploit the host immune system, which promotes antitumor activity. However, gemcitabine and 5-FU have been shown to activate NLRP3 in myeloid suppressor cells, leading to the production of IL-1, which reduces antitumor efficacy. Mechanistically, these agents destabilized the lysosome, releasing cathepsin B to activate NLRP3. IL-1 stimulated the production of IL-17 from CD4+ T cells, which in turn blunted the effectiveness of chemotherapy. Greater antitumor effects for both gemcitabine and 5-FU were observed when tumors were established in NLRP3'/' or Caps1 ^-/ ' mice or WT mice treated with IL-1RA. Consequently, activation of NLRP3 myeloid-derived suppressor cells limits the antitumor efficacy of gemcitabine and 5-FU (Bruchard, Melanie, et al. Chemotherapy-triggered cathepsin B release in myeloid-derived suppressor cells activates the Nlrp3 inflammasome and promotes tumor growth. Nature medicine 19.1 (2013): 57-64). Therefore, the compounds of the present invention may be useful in chemotherapy for the treatment of a number of cancers.

The compounds of the present invention, or pharmaceutically acceptable salts thereof, may be administered alone as monotherapy or may be administered in addition to one or more other agents and/or treatments. Such combination treatment can be carried out by simultaneous, sequential or separate doses of the individual components of therapy.

For example, therapeutic efficacy may be enhanced by administration of an adjuvant (ie, the adjuvant alone may have only minimal therapeutic benefit, but when combined with another therapeutic agent, the overall therapeutic benefit to the individual is increased). Alternatively, by way of example only, the benefit experienced by an individual may be increased by administering a compound of formula (I) with another therapeutic agent (which also includes a therapeutic regimen) that also has a therapeutic benefit.

In cases where the compound of the present invention is administered in combination with other therapeutic agents, the compound of the present invention may not require the same route of administration as other therapeutic agents and, due to different physical and chemical characteristics, may be entered in another way. For example, a compound of the present invention may be administered orally to generate and maintain good blood levels, while another therapeutic agent may be administered intravenously. Initial administration may be carried out in accordance with established protocols known in the art, and then, based on the observed effects, the skilled practitioner may adjust the dosage, routes of administration, and timing of administration.

- 13 044992

The specific choice of another therapeutic agent will depend on the diagnosis of the treating physicians and their judgment of the individual's condition and the appropriate treatment protocol. In accordance with this aspect of the invention, there is provided a combination for use in the treatment of a disease in which inflammasome activity is involved, comprising a compound of the present invention as defined above, or a pharmaceutically acceptable salt thereof, and another suitable agent.

In accordance with a further aspect of the invention there is provided a pharmaceutical composition which contains a compound of the invention or a pharmaceutically acceptable salt thereof in combination with a suitable pharmaceutically acceptable diluent or carrier.

Besides the use in therapeutic medicine? The compounds of formula (I) and their pharmaceutically acceptable salts can also be used as pharmacological agents in the development and standardization of in vitro and in vivo test systems for assessing the effects of inflammasome inhibitors in laboratory animals such as dogs, rabbits, monkeys, rats and mice , as part of the search for new therapeutic agents.

In any of the above pharmaceutical compositions, method, method, use, drug and manufacturing features of the present invention, any alternative embodiments of the macromolecules of the present invention described herein are also applicable.

Routes of administration.

The compounds of the invention, or pharmaceutical compositions containing these compounds, can be administered to a subject by any convenient route of administration, systemically/peripherally or locally (ie, at the site of desired action).

Routes of administration include, but are not limited to, oral (eg, oral administration); buccal; sublingual; transdermal (including, for example, bandage, patch, etc.); transmucosal (including, for example, bandage, patch, etc.); intranasal (for example, using a nasal spray); administration to the eyes (for example, eye drops); pulmonary (for example, by inhalation or insufflation therapy using, for example, an aerosol, for example, through the mouth or nose); rectal (for example, using a suppository or enema); vaginal (for example, with a pessary); parenteral, for example, by injection, including subcutaneous, intradermal, intramuscular, intravenous, intraarterial, intracardiac, intrathecal, intraspinal, intracapsular, subcapsular, intraorbital, intraperitoneal, intratracheal, subcuticular, intraarticular, subarachnoid and intrasternal; by implanting a depot or reservoir, for example, subcutaneously or intramuscularly.

In describing the present invention, the following examples are offered by way of illustration and not limitation.

Specific examples.

The invention will now be described with reference to the following illustrative examples. Some abbreviations that may appear in this section are defined as follows:

ACN - acetonitrile,

Boc - tert-butoxycarbonyl,

TFA - trifluoroacetic acid,

MeOH - methanol,

HCl - hydrochloric acid,

DCM - dichloromethane,

TLC - thin layer chromatography,

DMSO - dimethyl sulfoxide,

HPLC - high performance liquid chromatography,

EtOAc - ethyl acetate,

FCC - flash column chromatography,

THF - tetrahydrofuran,

NaOH - sodium hydroxide,

UPLC - ultra-performance liquid chromatography,

Ar - argon,

IV - starting material,

LC-MS - liquid chromatography-mass spectrometry,

Et ₃ N - triethylamine,

RM - reaction mixture, eq. - equivalents, k.t. - room temperature/ambient temperature, h - hours,

Pd ₂ (dba) ₃ - tris(dibenzylideneacetone)dipalladium(0),

Me ₄ tBuXPhos - methanesulfonato(2-di-tert-butylphosphino-3,4,5,6 tetramethyl-2',4',6'-triisopropyl- 14 044992

1,1-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(11),

HPLC - high performance liquid chromatography.

The compounds of the present invention can be prepared in accordance with the methods shown in the following schemes and examples using appropriate materials, and are further illustrated in the following specific examples. Analytical data for compounds obtained from the following examples are presented. Unless otherwise stated, all starting materials were obtained from commercial sources and used without further purification. Unless otherwise stated, all temperatures are expressed in °C and all reactions are carried out at room temperature. The compounds are typically purified by silica gel chromatography, preparative thin layer chromatography, or preparative HPLC.

1H NMR is recorded on 400 MHz spectrometers. Chemical shifts (δ) are presented in ppm. relative to the residual solvent signal (δ=2.5 ppm for ¹ H NMR in DMSO-d6). The ¹ H NMR data are presented as follows: chemical shift (multiplicity, binding constants and number of hydrogen atoms).

Multiplicity in abbreviated form is presented as follows: s (singlet), d (doublet), t (triplet) and m (multiplet).

LC-MS analyses.

UPLC-MS:

Equipment: Shimadzu LC-MS 2020 column: Waters Acquity UPLC HSS C18, 50 mm x 2.1 mm x 1.8 µm Eluents:

(A) 0.1% formic acid in ACN (B) 0.1% formic acid in water

Autosampler: injection volume: 1 µl

Pump:

Time [min] Flow [ml/min] %IN 0.00 0.5 95 0.00 0.5 95 4.00 0.5 5 5.00 0.5 5 5.20 0.5 95 6.00 0.5 95

Column compartment: column temperature: 25°C, analysis time: 6 min

Detector: Wavelength: 200-300 nm (254, 230, 270, 280 nm)

HPLC-MS:

Equipment: MS Bruker Amazon SL; LC Dionex Ultimate 3000; HPLC with UV-Vis detector or DAD column: Kinetex XB C18 4.6x50 mm 2.6 µm

Eluents:

(A) 0.1% formic acid-water solution (B) 0.1% formic acid-ACN solution Autosampler: injection volume: 1 µL

Column compartment: column temperature: 25°C, analysis time: 12 min Detector: wavelength 200-300 nm (220, 254, 280 nm)

- 15 044992

General methods.

General method A.

To a stirred solution of the amino ester (or amino ester hydrochloride with 1 eq. Et ₃ N) in ACN was added dropwise a solution of intermediate A in ACN. The reaction mixture was stirred overnight, then filtered. The resulting precipitate was washed with ACN and dried under reduced pressure to obtain the desired product. General method B.

Thionyl chloride (20 equiv.) was added dropwise to the methanol solution cooled to 0°C and the reaction mixture was stirred at 0°C for 30 minutes. The amino acid was added and the reaction mixture was stirred at ambient temperature overnight. The reaction mixture was evaporated under reduced pressure to obtain the desired product. General method C.

4 M HCl in dioxane (20 eq.) was added dropwise to a solution of Boc protected starting material in MeOH. The reaction mixture was stirred until no more starting material was observed by TLC and then evaporated to give the desired product.

Intermediate connections.

The intermediates below were prepared as follows: Intermediate A.

4-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (intermediate A).

Stage 3

Stage 1.

-Chloro-1 -(2,3-dihydro-1H-inden-5-yl)propan-1-one.

A suspension of aluminum chloride (12.4 g, 0.093 mol) in DCM (50 ml) under argon was cooled to -10°C with vigorous stirring. To this was added dropwise a solution of 3chloropropionyl chloride (11 g, 0.093 mol) and indan (10 g, 0.085 mol) in DCM (15 ml) over 0.5 h, the temperature was maintained between -15°C and -5° WITH. The reaction mixture was allowed to warm to room temperature and stirred overnight. The reaction mixture was added dropwise to cold (0°C) 2M HCl over 30 min, maintaining the temperature between 0°C and 10°C. The layers were separated and the aqueous phase was washed with DCM (3x30 ml). The combined organic layers were washed sequentially with water, saturated sodium bicarbonate solution and brine. The organic phases were dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure to about 30 ml. Hexane (50 ml) was added and evaporation continued, the procedure was repeated twice. After

- 16 044992 subsequent addition of hexane (50 ml) the slurry was filtered and dried to obtain 3-chloro-1-(2,3dihydro-1H-inden-5-yl)propan-1-one as a tan solid.

Yield=81%.

MS ES+: non-ionized.

1H NMR (400 MHz, DMSO-d6) δ 7.84 (d, 1H), 7.78-7.76 (m, 1H), 7.37 (d, J=8 Hz, 1H), 3.92 (t, J=6 Hz, 2H), 3.51 (t, J=6 Hz, 2H), 2.92 (t, J=8 Hz, 4H), 2.09-2.01 (m, 2H ).

Stage 2.

Mixture of 8-nitro-1,2,3,5,6,7-hexahydro-s-indacen-1-one, 4-nitro-1,2,3,5,6,7-hexahydro-8-indacen-1one and 5-nitro-1,2,3,6,7,8-hexahydro-s-indacene-3-one-3-chloro-1-(2,3-dihydro-1H-inden-5-yl)propan- 1-one (82 g, 0.39 mol) was added portionwise to concentrated sulfuric acid (71 ml, 1.34 mol). The resulting mixture was heated to a temperature of 60°C for 2 days. The reaction mixture was cooled to 0°C and a mixture of nitric acid (26 ml, 0.59 mol) and sulfuric acid (26 ml, 0.49 mol) was added dropwise. The reaction mixture was stirred at a temperature between 0°C and 5°C for 1 hour. The reaction mixture was slowly added to the water-DCM mixture while cooling in an ice bath. The layers were separated and the aqueous layer was extracted with DCM. The combined organic layers were washed successively with brine and saturated sodium bicarbonate. The organic layers were dried over Na ₂ SO ₄ and filtered. The crude mixture was purified using FCC (hexane/ethyl acetate). The products were then purified by crystallization from MeOH to obtain the desired products.

8-Nitro-1,2,3,5, 6,7-hexahydro-s-indacen-1-one.

Yield = 36%.

MS ES+: 218.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.67 (s, 1H), 3.15-3.08 (m, 2H), 3.04 (t, J=8 Hz, 2H), 2 .90 (t, J=8 Hz, 2H), 2.77-2.71 (m, 2H), 2.17-2.10 (m, 2H).

4-Nitro-1,2,3,5,6,7-hexahydro-s-ind acene-1-one.

Yield=5%.

MS ES+: 218.

1H NMR (400 MHz, DMSO- _d6 ) δ 7.82 (s, 1H), 3.41-3.36 (m, 2H), 3.34-3.29 (m, 3H), 3.02 (t, J=8 Hz, 2H), 2.77-2.69 (m, 2H), 2.17-2.10 (m, 2H).

5-Nitro-1,2,3,6,7,8-hexahydro-s-indacen-3-one.

Yield = 4%.

MS ES+: 218.

1H NMR (400 MHz, DMSO-d6) δ 8.09 (s, 1H), 3.39 (t, J=8 Hz, 2H), 3.14-3.09 (m, 2H), 3.01 (t, J=8 Hz, 2H), 2.81-2.73 (m, 2H), 2.23-2.15 (m, 2H).

Stage 3.

1,2,3,5,6,7-G hexahydro-s-indacene-4-amine.

A mixture of 8-nitro-1,2,3,5,6,7-hexahydro-s-indacen-1-one and 4-nitro-1,2,3,5,6,7-hexahydro-s-indacen-1-one (7.00 g, 0.032 mol) was suspended in MeOH (70 ml). Treated with 20% palladium hydroxide on carbon (50% wet, 1.72 g, 0.012 mol), then methanesulfonic acid (3.41 g, 0.035 mol). The mixture was hydrogenated at 35 psi for 5 hours. The catalyst was removed by filtration and washed with MeOH. The filtrate was diluted with water (350 ml) and then the pH was adjusted to 11 with 2N NaOH. The resulting slurry was filtered and the crude solids were recrystallized from MeOH/water (9:1) to give 1,2,3,5,6,7-hexahydro-s-indacene-4-amine as colorless crystalline needles.

Yield = 73%.

MS ES+: 174.1.

1H NMR (400 MHz, DMSO-d6) δ 6.35 (s, 1H), 4.52 (s, 2H), 2.72 (t, J=7 Hz, 4H), 2.59 (t, J =7 Hz, 4H), 2.00-1.93 (m, 4H).

Stage 4.

4-Isocyanato-1,2,3,5,6,7-hexahydro-s-indacene (intermediate A).

Into a stirred solution of 1,2,3,5,6,7-hexahydro-s-indacene-4-amine (1.1 g, 6.35 mmol) and Et ₃ N (0.973 ml, 6.98 mmol) in THF (20 ml) triphosgene (0.64 g, 2.16 mmol) was added in one portion. The mixture was heated to reflux for 4 hours, then cooled to room temperature. The THF was evaporated and the residue was treated with pentane and filtered through a pad of silica gel. Evaporation of the solvent in vacuo gave 4-isocyanato-1,2,3,5,6,7-hexahydro-s-indacene as a white solid.

Yield = 71%.

MS ES+: non-ionized.

1H NMR (400 MHz, chloroform-d) δ 6.96 (s, 1H), 2.94-2.89 (m, 8H), 2.22-2.03 (m, 4H).

Intermediate B.

Ethyl 1H,2H,3H,6H,7H,8H,9H-cyclopenta[a]naphthalene-5-amine (intermediate B).

- 17 044992

Stage 3

INTERMEDIATE CONNECTION B

Stage 1.

-Chloro-1-(5,6,7,8-tetrahydronaphthalene-2-yl)propan-1-one.

A suspension of aluminum chloride (5.58 g, 0.042 mol) in DCM (30 ml) under argon was cooled to -10°C with vigorous stirring. A solution of 3-chloropropionyl chloride (3.6 ml, 0.038 mol) and tetralin (5 g, 0.038 mol) in DCM (10 ml) was added dropwise over 0.5 h, the temperature was maintained between -15°C and - 5°C. The reaction mixture was allowed to warm to room temperature and stirred overnight. The reaction mixture was added dropwise to cold (0°C) 2M HCl over 30 min, maintaining the temperature between 0°C and 10°C. The layers were separated and the aqueous phase was washed with DCM (3x20 ml). The combined organic layers were washed sequentially with water, saturated sodium bicarbonate solution and brine. The organic phases were dried over Na ₂ SO ₄ , filtered and evaporated under reduced pressure to obtain 3-chloro-1(2,3-dihydro-1H-inden-5-yl)propan-1-one as a yellow solid.

Yield = 91%.

MS ES+: non-ionized.

¹ H NMR (400 MHz, DMSO-d6) δ 7.84 (d, 1H), 7.69-7.66 (m, 2H), 7.20 (d, J=8 Hz, 1H), 3, 91 (t, J=6 Hz, 2H), 3.49 (t, J=6 Hz, 2H), 2.78 (d, J=4 Hz, 4H), 1.77-1.72 (m, 4H).

Stage 2.

A mixture of 9-nitro-1H,2H,3H,5H,6H,7H,8H-cyclopenta[b]naphthalin-1-one, 4-nitro-1H,2H,3H,5H,6H,7H,8Hcyclopenta[b]naphthaline -1-one and 5-nitro-1H,2H,3H,6H,7H,8H,9H-cyclopenta[a]naphthalin-3-one.

3-Chloro-1-(5,6,7,8-tetrahydronaphthalene-2-yl)propan-1-one (7.52 g, 34 mmol) was added portionwise to concentrated sulfuric acid (36 ml). The resulting mixture was heated to a temperature of 60°C for 2 days. The reaction mixture was cooled to 0°C and a mixture of nitric acid (2.4 ml, 52 mmol) and sulfuric acid (2.4 ml) was added dropwise. The reaction mixture was stirred at a temperature between 0°C and 5°C for 1 hour. The reaction mixture was slowly added to the water-DCM mixture while cooling in an ice bath. The layers were separated and the aqueous layer was extracted with DCM. The combined organic layers were washed successively with brine and saturated sodium bicarbonate. The organic layers were dried over Na ₂ SO ₄ and filtered. The crude mixture was purified using FCC (hexane/ethyl acetate) to obtain a mixture of 9-nitro-III,2H,3H,5H,6H,7H,8Hcyclopenta[b]naphthalin-1-one, 4-nitro-1H,2H,3H, 5H,6H,7H,8H-cyclopenta[b]naphthalene-1-one and 5-nitro1H,2H,3H,6H,7H,8H,9H-cyclopenta[a]naphthalene-3-one in the form of a yellow semi-solid.

Yield = 13%.

MS ES+: 232.

Stage 3.

1H,2H,3H,6H,7H,8H,9H-cyclopenta[a]naphthaline-5-amine (intermediate B).

A mixture of 9-nitro-1H,2H,3H,5H,6H,7H,8H-cyclopentα[b]naphthalin-1-one, 4-nitro-1H,2H,3H,5H,6H,7H,8Hcyclopent[b] naphthalene-1-one and 5-nitro-1H,2H,3H,6H,7H,8H,9H-cyclopenta[a]naphthalene-3-one (0.992 g, 2.3 mmol) were suspended in MeOH (40 ml). Treat with 20% palladium hydroxide on carbon (50% wet, 0.389 g, 0.21 mmol), then methanesulfonic acid (0.32 ml, 4.8 mmol). The mixture was hydrogenated at 35 psi overnight. The catalyst was removed by filtration and washed with MeOH. The filtrate was diluted with water (50 ml) and then the pH was adjusted to 11 with 2M NaOH. The resulting slurry was filtered and the crude solids were purified with FCC (hexane/ethyl acetate) to give 1H,2H,3H,6H,7H,8H,9H-cyclopenta[a]naphthalene-5-amine as a brown oil.

Yield = 19%.

MS ES+: 188.4.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 6.35 (s, 1H), 4.42 (s, 2H), 2.69 (t, J=8 Hz, 2H), 2.58 (t , J=7 Hz, 2H), 2.48 (t, J=6 Hz, 2H), 2.32 (t, J=6 Hz, 2H), 1.95-1.88 (m, 2H), 1.76-1.60 (m, 4H).

Some intermediates identified herein may be new

- 18 044992 and they can be offered as an additional feature of the invention.

Additional source materials.

The starting materials for the preparation of the compounds of the present invention can be obtained by the methods described in the examples, or by methods known per se, as described in the literature on synthetic organic chemistry and known to the person skilled in the art, or can be obtained commercially. The starting materials for the processes can, if desired, also be formed in situ, without isolating them from the reaction mixture, but instead directly converting them further into the compounds of the invention or intermediates. On the other hand, as a rule, the reaction can be carried out in stages.

The following additional starting materials were used in the preparation of the compounds of the invention, and the process for their preparation is included below.

Methyl 2-amino-3-(2-hydroxyphenyl)propanoate hydrochloride.

IV: 2-amino-3-(2-hydroxyphenyl)propanoic acid.

General method B.

The product was used in the next step without further purification.

MS ES+: 196.

-[3-(Bromomethyl)phenyl]ethan-1-one.

A solution of meta-tolylethanone (5 g, 37 mmol), N-bromosuccinimide (1 eq., 6.6 g, 37 mmol) and benzoic acid peroxyanhydride (0.2 eq., 1.8 g, 7.5 mmol) in acetonitrile was stirred at a temperature of 85°C in an argon atmosphere overnight. The solvent was removed under reduced pressure. The residue was purified with FCC (0-5% EtOAc in hexane) to give the desired product as a yellow oil.

Yield = 58%.

MS ES+: non-ionized.

^1H NMR (400 MHz, chloroform-d) δ 8.00 (t, J=2 Hz, 1H), 7.94-7.88 (m, 1H), 7.65-7.59 (m, 1H ), 7.48 (t, J=8 Hz, 1H), 4.56 (s, 2H), 2.64 (s, 3H).

1,3-Diethyl 2-[(3-acetylphenyl)methyl]-2-acetamidopropanedioate.

Suspension of 1-[(3-bromomethyl)phenyl]ethan-1-one (2.5 g, 11.7 mmol), diethyl acetamidomalonate (1 eq., 2.55 g, 11.7 mmol), K2CO3 (1, 2 eq., 1.95 g, 14.1 mmol), potassium iodide (0.25 eq., 487 mg, 2.9 mmol) and Cs ₂ CO ₃ (1.2 eq., 4.59 g, 14 .1 mmol) in acetonitrile (100 ml) was heated to reflux and stirred overnight. The reaction mixture was cooled to room temperature, filtered through a pad of celite and concentrated in vacuo. Purification with FCC (0-50% EtOAc in hexane) gave the desired product as a white solid.

Yield = 67%.

MS ES+: 350.0.

3-(3-acetylphenyl)-2-aminopropanoic acid hydrochloride.

- 19 044992

A suspension of 1,3-diethyl 2-[(3-acetylphenyl)methyl]-2-acetamidopropanedioate (2.74 g, 7.84 mmol) in 6 M HCl (80 ml) was heated to reflux for 16 h. The reaction mixture was allowed to cool to room temperature. The solvent was evaporated and the solid was filtered, washed twice with diethyl ether and dried in vacuo to give the corresponding product as a white solid.

Yield = 98%.

MS ES ^- : 207.0.

Methyl 3-(3-acetylphenyl)-2-aminopropanoate.

IV: 3-(3-acetylphenyl)-2-aminopropanoic acid hydrochloride.

General method B.

The product was then purified with FCC (0-7% MeOH in DCM) to obtain the desired product as a white solid.

Yield = 8%.

MS ES+: 222.0.

Methyl (2R)-2-amino-3-(4-cyanophenyl)propanoate.

IV: (2R)-2-amino-3-(4-cyanophenyl)propanoic acid.

General method B.

The product was further partitioned into water pH=8-9 and EtOAc, separated and the organic products were dried (Na ₂ SO ₄ ) and concentrated. Subsequent purification with FCC (DCM/MeOH) gave the desired product.

Yield = 63%.

MS ES+: 205.

Methyl (2R)-2-amino-3-(3-cyanophenyl)propanoate hydrochloride.

IV: (2R)-2-amino-3-(3-cyanophenyl)propanoic acid.

General method B.

- 20 044992

Yield = 67%.

MS ES ⁺ : 205.

Methyl 2-amino-3-(3-bromophenyl)propanoate.

IV: 2-amino-3-(3-bromophenyl)propanoic acid.

General method B.

The product was further partitioned into pH 8-9 water and EtOAc, separated and the organics were dried (Na ₂ SO ₄ ) and concentrated to obtain the desired product.

Yield = 57%.

MSES ⁺ : 257.9; 259.9.

Methyl 3-(3-bromophenyl)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}propanoate.

IV: methyl 2-amino-3-(3-bromophenyl)propanoate.

General method A.

Yield = 74%.

MS ES ⁺ : 457;459.

Ή NMR (400 MHz, DMSO-I6) δ 7.83 (s, 1H), 7.47-7.42 (m, 1H), 7.42-7.38 (m, 1H), 7.27 ( t, J=8 Hz, 1H), 7.23-7.18 (m, 1H), 6.87 (s, 1H), 6.40 (d, J=8 Hz, 1H), 4.53- 4.48 (m, 1H), 3.66 (s, 3H), 3.11-3.07 (m, 1H), 2.992.93 (m, 1H), 2.79 (t, J=7 Hz , 4H), 2.63 (t, J=7 Hz, 4H), 1.98-1.91 (m, 4H).

Methyl (2R)-2-amino-3-(pyridin-3-yl)propanoate hydrochloride.

IV: 3-(3-pyridyl)-D-alanine.

General method B.

Yield = 63%.

MS ES ⁺ : 181.0.

Methyl 3-(3-bromophenyl)-2-{[(tert-butoxy)carbonyl]amino}propanoate.

Methyl 2-amino-3-(3-bromophenyl)propanoate hydrochloride (550 mg, 1.867 mmol) and Et ₃ N (0.520 ml, 1.2 eq., 3.734 mmol) were dissolved in dioxane (25 ml). A solution of di-tert-butyl dicarbonate (489 mg, 2 eq., 2.240 mmol) in dioxane (25 ml) was added dropwise. The reaction mixture was stirred at room temperature until the starting material was no longer observed by TLC. Raw

- 21 044992 the product was purified using FCC (hexane/EtOAc) to obtain the desired product.

Yield = 60%.

MS ES+: does not ionize.

Methyl 2-{[(tert-butoxy)carbonyl]amino}-3-[3-(1H-pyrazol-3-yl)phenyl]propanoate.

Methyl 3-(3-bromophenyl)-2-{[(tert-butoxy)carbonyl]amino}propanoate (100 mg, 0.28 mmol), (1Hpyrazol-3-yl)boronic acid (47 mg, 1.5 eq. ., 0.42 mmol), [1,1'bis(diphenylphosphino)ferrocene]dichloropalladium(P) (12 mg, 0.05 eq., 0.014 mmol) and Na ₂ CO ₃ (89 mg, 3 eq., 0.837 mmol) was dissolved in ACN (2 ml) with one drop of water. The reaction mixture was irradiated in a microwave reactor at 90°C for 1 hour. The crude reaction mixture was filtered through celite, washed with MeOH and concentrated under reduced pressure. The crude product was purified using FCC (DCM/MeOH) to obtain the desired product.

Yield = 26%.

MS ES+: 346.

Methyl 2-amino-3-[3-(1H-pyrazol-5-yl)phenyl]propanoate hydrochloride.

IV: methyl 2-{[(tert-butoxy)carbonyl]amino}-3-[3-(1H-pyrazol-3-yl)phenyl]propanoate.

General method C.

The crude product was used in the next step without further purification.

Methyl (2R)-2-amino-3-(3-hydroxyphenyl)propanoate hydrochloride.

IV: (2R)-2-amino-3-(3-hydroxyphenyl)propanoic acid.

General method B.

Yield = 74%.

MS ES+: 195.9.

(2R)-3-(3-Bromophenyl)-2-{[(tert-butoxy)carbonyl]amino}propanoate.

(2R)-3-(3-Bromophenyl)-2-{[(tert-butoxy)carbonyl]amino}propanoic acid (2.5 g, 7.26 mmol) and

- 22 044992

K ₂ CO ₃ (1.2 g, 1.2 eq., 8.72 mmol) was suspended in DMF (30 ml) and stirred at room temperature for 30 min, then the reaction mixture was cooled to 0°C. MeI (3.1 g, 3 eq., 21.8 mmol) was added dropwise, after completion of the reaction by TLC, water (150 mL) was added and the mixture was extracted with Et ₂ O. The organic phase was evaporated to give the desired product.

Yield = 88%.

MS ES+: non-ionized.

¹ H NMR (400 MHz, DMSO-d6) δ 7.46 (s, 1H), 7.43-7.40 (m, 1H), 7.33 (d, J=8 Hz, 1H), 7, 28-7.22 (m, 2H), 4.23-4.17 (m, 1H), 3.63 (s, 3H), 3.05-3.00 (m, 1H), 2.87- 2.81 (m, 1H), 1.33 (s, 9H).

Methyl (2R)-2-{[(tert-butoxy)carbonyl]amino}-3-[3-(1H-pyrazol-3-yl)phenyl]propanoate.

Methyl (2R)-3-(3-bromophenyl)-2-{[(tert-butoxy)carbonyl]amino}propanoate (300 mg, 0.84 mmol), (1H-pyrazol-3-yl)boronic acid (141 mg, 1.5 eq., 1.26 mmol), bis(diphenylphosphino)ferrocene]dichloropalladium(P) (34 mg, 0.05 eq., 0.042 mmol) and Na ₂ CO ₃ (266 mg, 3 eq., 2.51 mmol) was suspended in ACN (10 ml) and water (1 ml). The reaction mixture was irradiated in a microwave reactor at 90°C for 1 hour. The reaction mixture was filtered through Celite, washed with MeOH and concentrated under reduced pressure to obtain the desired product, used in the next step without purification.

Yield = 76%.

MS ES+: 246.

Methyl (2R)-2-amino-3-[3-(1H-pyrazol-5-yl)phenyl]propanoate hydrochloride.

IV: methyl (2R)-2-{[(tert-butoxy)carbonyl]amino}-3-[3-(1H-nirazol-3-yl)phenyl]nponanoate. General method C

The crude product was used in the next step without purification.

Methyl (2R)-2-amino-3-(3-bromophenyl)propanoate hydrochloride.

IV: methyl (2R)-3-(3-bromophenyl)-2-{[(tert-butoxy)carbonyl] amino} propanoate.

General method C.

The crude product was used in the next step without purification.

Yield = 97%.

MS ES+: 258; 260.

(2R)-3-(3-bromophenyl)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}propanoate.

- 23 044992

Br Br

IV: methyl (2R)-2-amino-3-(3-bromophenyl)propanoate hydrochloride.

General method A.

Yield = 89%.

MS ES+: 457;459.

(2R)-3-{[(tert-butoxy)carbonyl]amino}-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4yl)carbamoyl]amino}propanoate.

IV: methyl (2R)-2-amino-3-{[(tert-butoxy)carbonyl]amino}propanoate hydrochloride.

General method A.

Yield = 92%.

MS ES+: 418.

^1H NMR (400 MHz, DMSO-b6) δ 7.96 (s, 1H), 6.98 (t, J=6 Hz, 1H), 6.88 (s, 1H), 6.38 (d, J=8 Hz, 1H), 4.30-4.25 (m, 1H), 3.63 (s, 3H), 3.27 (t, J=6 Hz, 2H), 2.79 (t, J=7 Hz, 4H), 2.74-2.64 (m, 4H), 1.99-1.92 (m, 4H), 1.38 (s, 9H).

Methyl (2R)-2-{[(tert-butoxy)carbonyl] amino}-3-(3-acetamidophenyl)propanoate.

O

O

(2R)-3-(3-bromophenyl)-2-{[(tert-butoxy)carbonyl]amino}propanoate (50 mg, 0.14 mmol), K ₃ PO ₄ (62 mg, 0. 29 mmol, 2.1 eq.), Pd ₂ (dba) ₃ (6 mg, 0.007 mmol, 0.05 eq.) and Me ₄ tBuXPhos (17 mg, 0.035 mmol, 0.25 eq.). The tube was hermetically sealed, evacuated and filled with argon (three times). A solution of acetamide (17 mg, 2 eq., 0.28 mmol) in tert-butanol (5 ml) was added to the test tube. The reaction mixture was stirred at 110°C for 24 hours. The reaction mixture was filtered through Celite, washed with MeOH and concentrated under reduced pressure. The crude product was purified by FCC to obtain the desired product.

Yield = 27%.

MS ES+: 337.

Methyl (2R)-2-amino-3-(3-acetamidophenyl)propanoate hydrochloride.

IV: methyl (2R)-2-{[(tert-butoxy)carbonyl] amino}-3-(3-acetamidophenyl)propanoate.

- 24 044992

General method C.

The crude product was used in the next step without purification.

Methyl amino-3-(1-methyl-1H-pyrazol-4-yl)propanoate dihydrochloride.

IV: 2-amino-3-(1-methyl-1H-pyrazol-4-yl)propanoic acid.

General method B.

The product was used in the next step without purification.

Yield = 94%.

MS ES+: 184.

Methyl(2R)-2-{[(tert-butoxy)carbonyl]amino}-3-[3-(2-oxopyrrolidin-1-yl)phenyl]propanoate.

(2R)-3-(3-bromophenyl)-2-{[(tert-butoxy)carbonyl]amino}propanoate (50 mg, 0.14 mmol), K3PO4 (62 mg, 0.293 mmol, 2. 1 eq.), Pd ₂ (dba) ₃ (6 mg, 0.007 mmol, 0.05 eq.) and Me ₄ tBuXPhos (17 mg, 0.035 mmol, 0.25 eq.).

The tube was hermetically sealed, evacuated and filled with argon (three times). A solution of pyrrolidone (23 mg, 0.28 mmol, 2 eq.) in tert-butanol (5 ml) was added. The reaction mixture was stirred at 110°C for 24 hours. The reaction mixture was filtered through Celite, washed with MeOH and concentrated under reduced pressure to obtain the desired product, used in the next step without purification.

Yield = 59%.

MS ES+: 363.

Methyl (2R)-2-amino-3-{3-[(2-oxocyclopentyl)amino]phenyl}propanoate.

IV: methyl(2R)-2-{[(tert-butoxy)carbonyl]amino}-3-[3-(2-oxopyrrolidin-1-yl)phenyl]propanoate.

General method C.

The crude product was used in the next step without purification.

3-(1,2,3,5,6,7-hexαhydro-s-indacen-4-yl)-5-({3-[(1H-pyrazol-3-yl)amino]phenyl}methyl)imidazolidine2, 4-dion.

- 25 044992

Methyl (2R)-3-(3-bromophenyl)-2-{[(1,2,3,5,6,7hexahydro-s-indacen-4-yl)carbamoyl]amino}propanoate (75) was placed in a hermetically sealed test tube mg, 0.139 mmol), 3-aminopyrazole (14 mg, 0.164 mmol), tBuONa (33 mg, 0.293 mmol, 2.1 eq.), 2-di-tert-butylphosphino-2',4',6'triisopropylbiphenyl, tBuXPhos ligand (4 mg, 0.008 mmol, 0.05 eq.) and chloro[2-(di-tert-butylphosphino)2',4',6'-triisopropyl-1,G-biphenyl][2-(2- aminoethyl)phenyl)]palladium(II) tBuXPhos 1G precatalyst (6 mg, 0.008 mmol, 0.05 eq.) The tube was sealed, evacuated and filled with argon (three times) and then tert-butanol (2 ml) was added. The reaction mixture was stirred at 110°C for 24 hours. The reaction mixture was filtered through Celite, washed with MeOH and concentrated under reduced pressure. The crude product was purified by FCC to obtain the desired product.

Yield = 85%.

MS ES+: 427.

(2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-{3-[(1H-pyrazol-3yl)amino ]phenyl} propanoic acid.

3-(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)-5 -({3-[(1 H-pyrazol-3-yl)amino]phenyl}methyl)imidazolidine2 ,4-dione (60 mg, 0.141 mmol) was suspended in 5 M NaOH (2 ml) and stirred at room temperature overnight. The reaction mixture was evaporated to obtain the desired product, which was used in the next step without purification.

Yield = 80%.

MS ES+: 446.4.

(2R)-2-Amino-3-[3-(1H-pyrazol-3-yl)phenyl]propanoic acid dihydrochloride.

A mixture of methyl (2R)-2-{[(tert-butoxy)carbonyl]amino}-3-[3-(1H-pyrazol-3-yl)phenyl]propanoate (203 mg, 0.60 mmol) and 6 M HCl ( 10 ml) was heated at reflux overnight. The reaction mixture was allowed to cool to room temperature, diluted with water (50 ml) and washed with diethyl ether (50 ml). The aqueous phase was then evaporated under reduced pressure to obtain the desired product as a yellow solid.

Output = 100%.

MS ES+: 232.1.

^1H NMR (400 MHz, DMSO-d6) δ 8.59-8.43 (m, 3H), 7.80 (d, J=2 Hz, 1H), 7.79-7.77 (m, 1H ), 7.76-7.72 (m, 1H), 7.39 (t, J=8 Hz, 1H), 7.27-7.21 (m, 1H), 6.75 (d, J= 2 Hz, 1H), 4.25-4.17 (m, 1H), 3.22-3.17 (m, 2H).

2-{[(1,2,3,5,6,7-G hexahydro-s-indacen-4-yl)carbamoyl] amino} acetic acid.

Methyl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}acetate (140 mg, 0.5 mmol) was suspended in MeOH (1 ml). 1M NaOH (5 ml) was added and the reaction mixture was stirred at room temperature overnight, then concentrated in vacuo. The residue was acidified to pH 2 with 2M HCl and the resulting residue was filtered and washed with water to obtain the desired product as a white solid.

- 26 044992

Yield = 73%.

MS ES ⁺ : 275.0.

^1H NMR (400 MHz, DMSO) δ 12.47 (s, 1H), 7.90 (s, 1H), 6.88 (s, 1H), 6.26 (t, J=6 Hz, 1H) , 3.76 (d, J=6

Hz, 2H), 2.80 (t, J=7 Hz, 4H), 2.70 (t, J=7 Hz, 4H), 2.01-1.91 (m, 4H).

Methyl (2R)-2-amino-3-methoxypropanoate hydrochloride.

IV: (2R)-2-amino-3-methoxypropanoic acid.

General method B.

The product was used in the next step without further purification.

MS ES+: 175 [M+ACN].

2-{[(tert-Butoxy)carbonyl]amino}-3-[3-( 1 H-pyrazol-3-yl)phenyl]propanoic acid.

reactor was charged with (2R)-2-[(tert-butoxy)carbonylamino]-3-(3-bromophenyl)propanoic acid (1.6 g, 4.66 mmol, 1 eq.), 1H-pyrazole-3- boronic acid (1.3 g, 3.6 mmol, 2.5 eq.), Na ₂ CO ₃ (618 mg, 5.83 mmol, 4 eq.) and MeCN:H ₂ O (10:1, 22 ml ). The reaction mixture was purged with argon and Pd(dppf)Cl ₂ (341 mg, 0.46 mmol, 0.1 eq.) was added. The reaction mixture was heated at 90° C. under microwave irradiation for 1 hour. Then filtered through a pad of celite, washed with MeOH and the filtrate was concentrated in vacuo to give the title compound as a brown solid.

Yield = 51%.

MS ES+: 332.2.

Ethyl (2R)-2-amino-3-[3-(1H-pyrazol-3-yl)phenyl]propanoate hydrochloride.

2-{[(tert-Butoxy)carbonyl]amino}-3-[3-(1H-pyrazol-3-yl)phenyl]propanoic acid (250 mg, 1 mmol, 1 eq.) was dissolved in EtOH and the reaction mixture was cooled to a temperature of 0°C. Thionyl chloride (0.029 mL, 0.83 mmol, 1.1 eq.) was added and the reaction mixture was stirred at 70° C. overnight. After cooling to room temperature, the mixture was concentrated in vacuo. Et ₂ O was added and the resulting brown solid was filtered. The solid was dissolved in MeOH and filtered through an SCX cartridge, washing with MeOH and eluting the product with 1 M NH3 in MeOH. The filtrate was evaporated to give the title compound as a pale brown solid.

Yield = 37%.

MS ES+: 260.2.

2-methoxyethyl (2R)-2-amino-3-(pyridin-3-yl)propanoate hydrochloride.

- 27 044992

Thionyl chloride (21 µl, 1.1 eq.). The reaction mixture was heated at 60°C for 2 hours in an argon atmosphere. The reaction mixture was then allowed to cool to room temperature, poured into saturated aqueous NaHCO ₃ and the mixture was extracted twice with DCM. The combined organics were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title compound as a white solid.

Yield = 71%.

MS ES+: 225.3.

¹ H NMR (400 MHz, DMSO-d6) δ 8.42 (d, J=2 Hz, 1H), 8.41 (d, J=2 Hz, 1H), 7.68-7.58 (m, 1H), 7.33-7.27 (m, 1H), 4.15-4.11 (m, 1H), 3.63-3.59 (m, 1H), 3.53-3.44 ( m, 3H), 3.40-3.28 (m, 2H), 3.25 (s, 3H), 2.92-2.85 (m, 1H), 2.84-2.76 (m, 1H).

Cyclobutyl (2R)-2-amino-3-(pyridin-3-yl)propanoate.

(2R)-2-amino-3-(pyridin-3-yl)propanoic acid (100 mg, 0.60 mmol, 1 eq.) and cyclobutanol (860 mg, 12.03 mmol, 20 eq.) were suspended in toluene. Para-toluenesulfonic acid monohydrate (343 mg, 1.80 mmol, 3 eq.) was added and the mixture was heated to reflux for 2 hours. The reaction mixture was concentrated under reduced pressure and the residue was dissolved in DCM/water (1:1). The mixture was neutralized with saturated aqueous NaHCO ₃ and the aqueous layer was extracted twice with DCM. The combined organics were washed with brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title compound as a brown oil.

Yield = 46%.

MS ES+: 222.3.

(2R)-2-{[(tert-Butoxy)carbonyl]amino}-3-(pyridin-3-yl)propanoic acid.

(2R)-2-Amino-3-(pyridin-3-yl)propanoic acid (1.5 g, 9 mmol, 1 eq.) was dissolved in dioxane (30 ml) and water (30 ml), then the resulting solution was treated sodium bicarbonate (3 g, 36.1 mmol, 4 eq.). The resulting mixture was cooled to 0°C and a solution of Boc anhydride (2.36 g, 11 mmol, 1.2 eq.) in dioxane (10 ml) was added dropwise. The reaction mixture was stirred at 0°C for 1 hour and allowed to warm to room temperature overnight. The dioxane was evaporated under reduced pressure and the resulting aqueous solution was washed twice with ethyl acetate. The aqueous layer was neutralized with 10% aqueous potassium bisulfate and the solution was extracted three times with n-butanol. The combined organic layers were dried over sodium sulfate, filtered and evaporated in vacuo to give the title compound as a light yellow oil.

Yield = 51%.

MS ES+: 267.2.

^1H NMR (400 MHz, DMSO-Y6) δ 12.74 (s, 1H), 8.46-8.43 (m, 1H), 8.43-8.39 (m, 1H), 7.67 (d, J=8 Hz, 1H), 7.35-7.28 (m, 1H), 7.17 (d, J=8 Hz, 1H), 4.16-4.08 (m, 1H) , 3.10-3.02 (m, 1H), 2.87-2.78 (m, 1H), 1.31 (s, 9H).

Cyclopropylmethyl (2R)-2-{[(tert-butoxy)carbonyl]amino}-3-(pyridin-3-yl)propanoate.

- 28 044992

(2R)-2-{[(tert-butoxy)carbonyl]amino}-3-(pyridin-3-yl)propanoic acid (300 mg, 1.12 mmol, 1 eq.) and DMAP (14 mg, 0.113 mmol , 0.1 eq.) was dissolved in dry DCM (12 ml). The reaction mixture was cooled to 0°C and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (281 mg, 1.46 mmol, 1.3 eq.) was added, followed by cyclopropylmethanol (119 μl, 1.46 mmol, 1.3 eq.). The reaction mixture was stirred at room temperature under argon for 18 hours. Ethyl acetate was added and the insoluble material was filtered off. The filtrate was concentrated in vacuo to give the title compound as an oil.

Yield = 56%.

MS ES+: 321.3.

Cyclopropylmethyl (2R)-2-amino-3-(pyridin-3-yl)propanoate.

Cyclopropylmethyl (2R)-2-{[(tert-butoxy)carbonyl]amino}-3-(pyridin-3-yl)propanoate (200 mg, 0.52 mmol) was dissolved in DCM (1 ml) and TFA (2 ml ). The reaction mixture was stirred at room temperature overnight. Then diluted with DCM and neutralized with saturated aqueous NaHCO ₃ solution. The aqueous layer was extracted twice with DCM and the combined organics were dried over Na ₂ SO ₄ , filtered and concentrated in vacuo to give the title compound as a yellow oil.

Yield = 59%.

MS ES+: 221.3.

Cyclopentyl (2R)-2-{[(tert-butoxy)carbonyl] amino }-3 -(pyridin-3-yl)propanoate.

To a solution of (2K)-2-{[(tert-butoxy)carbonyl]amino}-3-(pyridin-3-yl)propanoic acid (150 mg, 0.56 mmol, 1 eq.) in DMF (4 ml) at 0°C, cyclopentanol (256 μl, 2.81 mmol, 5 eq.), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (140 mg, 0.73 mmol, 1.3 eq.) and DMAP (7 mg, 0.056 mmol, 0.1 eq.). The reaction mixture was stirred at room temperature overnight, then diluted with ethyl acetate. The mixture was filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by FCC (NH2 modified silica gel) with hexane:EtOAc (4:1) to give the title compound as a colorless oil.

Yield = 31%.

MS ES+: 335.3.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.45-8.40 (m, 2H), 7.67 (d, J=8 Hz, 1H), 7.37-7.29 (m, 2H), 5.10-5.02 (m, 1H), 4.17-4.08 (m, 1H), 3.03-2.96 (m, 1H), 2.93-2.86 ( m, 1H), 1.85-1.70 (m, 2H), 1.65-1.55 (m, 3H), 1.53-1.40 (m, 3H), 1.33 (s, 9H).

Di-TFA salt of cyclopentyl (2R)-2-amino-3-(pyridin-3-yl)propanoate.

Cyclopentyl (2R)-2-{[(tert-butoxy)carbonyl]amino}-3-(pyridin-3-yl)propanoate (100 mg, 0.26 mmol,

- 29 044992 eq.) was dissolved in DCM (1 ml) and TFA (2 ml) was added. The reaction mixture was stirred overnight at room temperature and concentrated under reduced pressure to give the title compound as a brown oil.

Yield = 78%.

MS ES+: 235.3.

(2R)-2-Amino-3-(pyridin-3-yl)propanoic acid hydrochloride.

In a pressure vessel, (2R)-2-amino-3-(pyridin-3-yl)propanoic acid (2 g, 12.0 mmol, 1 eq.) was suspended in dry EtOH (30 ml). The mixture was cooled to 0°C in an ice bath and conc. sulfuric acid (0.5 ml). The vessel was sealed and the mixture was stirred at 85°C for 18 hours. After being allowed to cool to room temperature, the reaction mixture was evaporated to one quarter of its volume and poured into sat. NaHCO ₃ solution. The mixture was extracted four times with CHCl ₃ and the combined organic layers were dried over sodium sulfate and filtered. 4M HCl in dioxane (12 ml) was added to the filtrate. The resulting solution was evaporated to give a colorless oil, which was dissolved in EtOH (10 ml). The solution was added to vigorously stirred Et ₂ O (100 ml) and stirring was continued for 1 hour until the resulting oil solidified into a white solid. The solid was filtered, washed with Et ₂ O and dried in vacuo to give the title compound as a white powder.

Yield = 61%.

MS ES+: 195.3.

¹ H NMR (400 MHz, DMSO-d6) δ 9.11-8.91 (m, 4H), 8.86 (d, J=6 Hz, 1H), 8.56 (d, J=8 Hz, 1H), 8.08-8.00 (m, 1H), 4.55-4.45 (m, 1H), 4.27-4.10 (m, 2H), 3.47 (d, J= 7 Hz, 2H), 1.17 (t, J=7 Hz, 2H).

Methyl 2-amino-3-(4-methyl-1H-pyrazol-1-yl)propanoate hydrochloride.

A solution of 2-amino-3-(4-methyl-1H-pyrazol-1-yl)propanoate (70 mg, 0.34 mmol) in 3M hydrochloric acid in methanol (5 ml) was stirred at room temperature for 16 hours. Solvent removed in vacuo to obtain the desired product.

Yield = 92%.

MS ES+: 184.

Ethyl 2-amino-3-(pyrimidin-2-yl)propanoate dihydrochloride.

In a vessel, 2-amino-3-(pyrimidin-2-yl)propanoic acid dihydrochloride (0.10 g, 0.417 mmol) was suspended in EtOH (1 ml) and cooled to 0°C. Added conc. H2SO4 (0.1 ml), the vessel was hermetically sealed and heated at 80°C for 18 hours. The reaction mixture was poured into sat. aqueous NaHCO ₃ solution and extracted four times with CHCl ₃ . The combined organics were dried over Na ₂ SO ₄ and filtered. 4M HCl in dioxane (2 ml) was added to the filtrate with stirring. This solution was evaporated to give a colorless oil, which was then dissolved in a minimal amount of EtOH (approx. 2 ml). The solution was added to rapidly stirred MeCN (10 ml) to crystallize the product. The solid was filtered and dried under vacuum to obtain the desired product as an off-white solid.

Yield = 66%.

- 30 044992

MS ES ⁺ : 196.3.

Methyl (2R)-2-amino-3-(5-methoxypyridin-3-yl)propanoate dihydrochloride.

Methyl (2R)-2-{[(tert-butoxy)carbonyl]amino}-3-(5-methoxypyridin-3-yl)propanoate (0.26 g, 0.8 mmol) was dissolved in 4 M HCl in dioxane (5 ml) and stirred at room temperature for 4 hours. The solvent was evaporated and the solid was dissolved in water and washed with EtOAc. The aqueous phase was freeze-dried to obtain the desired product as a brown solid.

Yield = 37%.

MS ES ⁺ : 211.2.

Ethyl 2-[(diphenylmethylidene)amino] acetate.

Glycine ethyl ester hydrochloride (1.00 g, 7.16 mmol) was dissolved in dry DCM (40 ml). Benzophenone imine (1.20 mL, 7.16 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 18 hours. The reaction mixture was filtered through Celite, washed with DCM and the filtrate was concentrated in vacuo. The resulting oil was triturated in hexane to obtain the desired product as a white solid.

Yield = 97%.

MS ES ⁺ : 268.

Ethyl 2-[(diphenylmethylidene)amino]-3 -(3-methyl-1,2,4-oxadiazol-5-yl)propanoate.

In a dry flask under nitrogen atmosphere, 2M LDA in THF (0.59 mL, 1.18 mmol) was cooled to 78°C. A solution of ethyl 2-[(diphenylmethylidene)amino]acetate in THF (12 ml) was added and the reaction mixture was stirred at -78°C for 30 minutes. 5-(chloromethyl)-3-methyl1,2,4-oxadiazole (0.12 mL, 1.18 mmol) was added dropwise and the reaction mixture was stirred at -78°C for 1 hour, then at room temperature for 18 hours. The reaction mixture was quenched with sat. NH ₄ Cl solution, diluted with water and extracted with EtOAc. Organic products were dried over Na ₂ SO ₄ , filtered and evaporated. The crude was purified using FCC (silica gel, 20% EtOAc in hexane) to obtain the desired product.

Yield = 20%.

MS ES ⁺ : 364.

Ethyl 2-amino-3-(3-methyl-1,2,4-oxadiazol-5-yl)propanoate hydrochloride.

Ethyl 2-[(diphenylmethylidene)amino]-3-(3-methyl-1,2,4-oxadiazol-5-yl)propanoate (80 mg, 0.23 mmol) was dissolved in diethyl ether (2 ml) and cooled to temperature 0°C. 1M aqueous HCl (1.1 mL, 1.1 mmol) was added dropwise and the reaction mixture was allowed to warm to room temperature. The reaction mixture was stirred for 72 hours and then concentrated to remove the organic solvent. The aqueous phase was diluted with 1M HCl and washed with Et ₂ O. The aqueous phase was concentrated in vacuo and freeze dried to obtain the desired product as a yellow solid.

Yield = 80%.

MS ES+: 200.

Methyl (2R)-2-{[(tert-butoxy)carbonyl] amino}-3-(pyridazin-3-yl)propanoate.

Zinc powder (0.12 g, 1.8 mmol) was added to a dry flask purged with nitrogen. Dry DMF (1.0 ml) was added followed by iodine (43 mg, 0.2 mmol). The color of the solution changed from colorless to yellow and then returned to colorless. Methyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-iodopropanoate (0.20 g, 0.60 mmol) was added, followed by iodine (43 mg, 0.2 mmol). The solution was stirred at ambient temperature with observed heat evolution. To this solution were added Pd ₂ (dba) ₃ (28 mg, 0.04 mmol), SPhos (25 mg, 0.12 mmol) and 3-bromopyridazine (0.25 g, 1.6 mmol). The reaction mixture was stirred at room temperature under nitrogen atmosphere for 18 hours. The reaction mixture was filtered twice and purified using FCC (silica gel, EtOAc/hexane) to obtain the desired product.

Yield = 43%.

MS ES+: 282.

Methyl (2R)-2-amino-3-(pyridazin-3-yl)propanoate hydrochloride.

General method C.

Yield = 97%.

MS ES+: 182.

Ethyl 2-[(diphenylmethylidene)amino]-3-(1,2-oxazol-4-yl)propanoate.

The dry flask was filled with 2M LDA in THF/hexane/toluene (0.59 mL, 1.18 mmol) and cooled to -78°C under nitrogen. A solution of ethyl 2[(diphenylmethylidene)amino]acetate (0.30 g, 1.12 mmol) in dry THF (16 ml) was added dropwise. The reaction mixture was stirred at -78°C for 30 min, then 4-(bromomethyl)-1,2-oxazole (0.19 g, 1.18 mmol) was added and stirring was continued at -78°C for 1 hours. The reaction mixture was allowed to warm to room temperature and stirred for 16 hours. The reaction mixture was cooled on ice water and quenched with sat. NH4Cl solution. Organic products were extracted with EtOAc, dried over sodium sulfate, filtered and concentrated. The desired product was obtained and used directly without further purification.

Yield = 51%.

MS ES+: 349.1.

Ethyl 2-amino-3-(1,2-oxazol-4-yl)propanoate hydrochloride.

- 32 044992

Ethyl 2-[(diphenylmethylidene)amino]-3-(1,2-oxazol-4-yl)propanoate (0.30 g, 0.86 mmol) was dissolved in diethyl ether (4 ml) and cooled to 0°C . 1M hydrochloric acid (2.0 mL, 1.0 mmol) was added dropwise and the reaction mixture was allowed to warm to room temperature and stirred for 16 hours. The reaction mixture was diluted with water and washed with diethyl ether. The aqueous phase was dried under vacuum to obtain the desired product, used as such.

Yield = 94%.

MS ES+: 185.2.

Ethyl 2-[(diphenylmethylidene)amino]-3-(1,2-oxazol-3-yl)propanoate.

The dry flask was filled with 2M LDA in THF/hexane/toluene (0.78 mL, 1.56 mmol) and cooled to -78°C under nitrogen. A solution of ethyl 2-[(diphenylmethylidene)amino]acetate (0.40 g, 1.49 mmol) in dry THF (14 ml) was added dropwise. The reaction mixture was stirred at 78°C for 30 minutes, then 3-(bromomethyl)isoxazole (0.148 ml, 1.56 mmol) was added and stirring was continued at -78°C for 1 hour. The reaction mixture was allowed to warm to room temperature. temperature and stirred for 16 hours. The reaction mixture was cooled on ice water and quenched with sat. NH ₄ Cl solution. Organic products were extracted with EtOAc, dried over sodium sulfate, filtered and concentrated. The desired product was obtained as a yellow oil and used directly without further purification.

Yield = 25%.

MS ES+: 349.1.

Ethyl 2-amino-3-(1,2-oxazol-3-yl)propanoate hydrochloride.

Ethyl 2-[(diphenylmethylidene)amino]-3-(1,2-oxazol-3-yl)propanoate (0.13 g, 0.39 mmol) was dissolved in diethyl ether (2 ml) and cooled to 0°C . 1M hydrochloric acid (1.9 mL, 1.9 mmol) was added dropwise; the reaction mixture was allowed to warm to room temperature and stirred for 16 hours. The reaction mixture was diluted with 1M hydrochloric acid and washed with diethyl ether. The aqueous phase was dried under vacuum to obtain the desired product as a yellow solid, used as such.

Yield = 81%.

MS ES+: 185.

6-(Prop-1-en-2-yl)-2,3-dihydro-1H-inden-5-amine.

6-Bromo-2,3-dihydro-1H-inden-5-amine (1.50 g, 7.1 mmol) and K3PO4 (3.75 g, 17.7 mmol) were placed in a test tube. Toluene (12 ml) and water (6 ml) were added. Palladium acetate (0.16 g, 0.7 mmol), tricyclohexylphosphine (0.20 g, 0.7 mmol) and isopropenyl boronic acid pinacol ester (1.78 g, 10.6 mmol) were then added, the tube was sealed and heated at 105°C for 16 hours. The reaction mixture was filtered through celite, the organic solvent was evaporated, and the resulting suspension was partitioned between EtOAc and brine. The organic phase was concentrated and purified using FCC (silica gel, 4:1 hexane/EtOAc) to obtain the desired product.

Yield = 5%.

MS ES+: 174.3.

6-(Propan-2-yl)-2,3-dihydro-1H-ind en-5-amine.

- 33 044992

6-(Prop-1-en-2-yl)-2,3-dihydro-1H-inden-5-amine (0.267 g, 1.54 mmol) was dissolved in MeOH (5 ml). Added 10% moisture. Pd/C (16 mg) and the reaction mixture was purged with argon. The reaction mixture was hydrogenated using a Parr hydrogenation apparatus for 6 hours. The reaction mixture was filtered through celite and concentrated to give the desired product, which was used without further purification.

Yield = 82%.

MS ES+: 176.3.

5-Isocyanato-6-(propan-2-yl)-2,3-dihydro-1H-indene.

6-(Propan-2-yl)-2,3-dihydro-1H-inden-5-amine (222 mg, 1.27 mmol) was dissolved in THF (10 ml) and triethylamine (0.19 ml, 0. 14 mmol). The reaction mixture was then treated with triphosgene (128 mg, 0.4 mmol), the reaction mixture was refluxed for 4 hours and cooled to room temperature. The solvent was removed in vacuo, the residue was dissolved in pentane and filtered through silica gel. The filtrate was evaporated to obtain the desired product.

Yield = 26%.

MS ES+ in MeOH: 234.3 (carbamate).

Methyl (2R)-2-{[(tert-butoxy)carbonyl]αmino}-3-(pyrimidin-5-yl)propanoate.

Zinc powder (0.12 g, 1.8 mmol) was added to a dry flask purged with nitrogen. Dry DMF (1.0 ml) was added followed by iodine (43 mg, 0.2 mmol). The color of the solution changed from colorless to yellow and then returned to colorless. Methyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-iodopropanoate (0.20 g, 0.60 mmol) was added, followed by iodine (43 mg, 0.2 mmol). The solution was stirred at ambient temperature with observed heat evolution. To this solution were added Pd ₂ (dba) ₃ (28 mg, 0.04 mmol), SPhos (24 mg, 0.12 mmol) and 5-iodopyrimidine (0.33 g, 1.6 mmol). The reaction mixture was stirred at room temperature under nitrogen atmosphere for 18 hours. The reaction mixture was filtered twice and then purified using FCC (silica gel, EtOAc/hexane) to obtain the desired product.

Yield = 80%.

MS ES+: 282.

Methyl (2R)-2-amino-3-(pyrimidin-5-yl)propanoate hydrochloride.

General method C.

Yield = 94%.

MS ES+: 182.2.

Methyl (2R)-2-{[(tert-butoxy)carbonyl] amino}-3-(pyrazin-2-yl)propanoate.

- 34 044992

Oh oh

Zinc powder (0.24 g, 3.6 mmol) was added to a dry flask purged with nitrogen. Dry DMF (2 ml) was added followed by iodine (86 mg, 0.4 mmol). The color of the solution changed from colorless to yellow and then returned to colorless. Methyl (2S)-2-{[(tert-butoxy)carbonyl]amino}-3-iodopropanoate (0.40 g, 1.2 mmol) was added, followed by iodine (86 mg, 0.4 mmol). The solution was stirred at ambient temperature with observed heat evolution. To this solution were added Pd ₂ (dba) ₃ (28 mg, 0.04 mmol), SPhos (24 mg, 0.12 mmol) and 2-iodopyrazine (0.32 g, 1.5 mmol). The reaction mixture was stirred at room temperature under nitrogen atmosphere for 18 hours. The reaction mixture was filtered twice and then purified using FCC (silica gel, EtOAc/hexane) to obtain the desired product.

Yield = 90%.

MS ES+: 282.

Methyl (2R)-2-amino-3-(pyrazin-2-yl)propanoate hydrochloride.

General method C.

Yield = 66%.

MS ES+: 182.1.

Ethyl 2-[(diphenylmethylidene)amino]-3-(pyridazin-4-yl)propanoate.

The dry flask was filled with 2M LDA in THF/hexane/toluene (1.2 mL, 4.86 mmol) and cooled to -78°C under nitrogen. A solution of ethyl 2[(diphenylmethylidene)amino]acetate (0.65 g, 2.43 mmol) in dry THF (25 ml) was added dropwise. The reaction mixture was stirred at -78°C for 30 minutes, then 4(bromomethyl)pyridazine hydrobromide (0.65 g, 2.55 mmol) and triethylamine (0.356 ml, 2.55 mmol) were added and stirring was continued at - 78°C for 1 hour. The reaction mixture was allowed to warm to room temperature and stirred for 16 hours. The reaction mixture was cooled in ice water and quenched with sat. NH ₄ Cl solution. Organic products were extracted with EtOAc, dried over sodium sulfate, filtered and concentrated. The crude product was purified using FCC (silica gel, 20% (EtOAc+1% Et ₃ N) in hexane to obtain the desired product.

Yield = 13%.

MS ES ⁺ : 360.1.

Ethyl 2-amino-3-(pyridazin-4-yl)propanoate hydrochloride.

Ethyl 2-[(diphenylmethylidene)amino]-3-(pyridazin-4-yl)propanoate (0.14 g, 0.38 mmol) was dissolved in diethyl ether (2.5 ml). 1M hydrochloric acid (1.0 ml, 1.0 mmol) was added and the reaction mixture was stirred for 16 hours. The reaction mixture was diluted with 1M hydrochloric acid and washed with diethyl ether. The aqueous phase was dried under vacuum to obtain the desired product as a brown oil, used as such.

Yield = 83%.

- 35 044992

MS ES ⁺ : 196.

(Pyrimidin-4-yl)methyl methanesulfonate.

A solution of (pyrimidin-4-yl)methanol (0.20 g, 1.82 mmol) in DCM (4 ml) was cooled to 0°C and treated with triethylamine (0.506 ml, 3.63 mmol) and methanesulfonic acid (0.281 ml , 3.63 mmol). The reaction mixture was allowed to warm to room temperature and stirred for 4 hours. The reaction mixture was diluted with DCM, washed successively with water and brine, dried over sodium sulfate and concentrated to give the desired product, used directly.

Yield = 91%.

MS ES ⁺ : 188.9.

Ethyl 2-[(diphenylmethylidene)amino]-3-(pyrimidin-4-yl)propanoate.

The dry flask was filled with 2M LDA in THF/hexane/toluene (2.25 mL, 4.50 mmol) and cooled to -78°C under nitrogen. A solution of ethyl 2[(diphenylmethylidene)amino]acetate (0.60 g, 2.24 mmol) in dry THF (20 ml) was added dropwise. The reaction mixture was stirred at -78°C for 30 minutes, then (pyrimidin-4-yl)methyl methanesulfonate (0.44 g, 2.36 mmol) was added and stirring was continued for 1 hour at -78°C. The reaction mixture was allowed to warm to room temperature and stirred for 16 hours. The reaction mixture was cooled on ice water and quenched with sat. NH ₄ Cl solution. The organic products were extracted with EtOAc, dried over sodium sulfate, filtered and concentrated to obtain the desired product, used directly in the next step.

Yield = 14%.

MS ES ⁺ : 360.1.

Ethyl 2-amino-3-(pyrimidin-4-yl)propanoate hydrochloride.

Ethyl 2-[(diphenylmethylidene)amino]-3-(pyrimidin-4-yl)propanoate (0.46 g, 1.71 mmol) was dissolved in diethyl ether (3 ml). 1M hydrochloric acid (3 ml, 3 mmol) was added and the reaction mixture was stirred for 16 hours. The reaction mixture was diluted with 1M hydrochloric acid and washed with diethyl ether. The aqueous phase was dried under vacuum to obtain the desired product, used as such.

Yield = 25%.

MS ES ⁺ : 196.

Examples of compounds according to the invention.

The following compounds of the invention were prepared as follows, with the necessary steps and starting materials required as previously described.

2A

Methyl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(3-hydroxyphenyl)propanoate.

- 36 044992

IV: methyl 2-amino-3-(3-hydroxyphenyl)propanoate hydrochloride.

General method A.

Yield = 8%.

MS ES+: 395.2.

1H NMR (400 MHz, DMSO-b ₆ ) δ 9.31 (s, 1H), 7.88 (s, 1H), 7.08 (t, J=8 Hz, 1H), 6.87 (s, 1H), 6.67-6.61 (m, 1H), 6.61-6.55 (m, 2H), 6.29 (d, J=8 Hz, 1H), 4.48-4.43 (m, 1H), 3.64 (s, 3H), 2.99-2.83 (m, 2H), 2.79 (t, J=7 Hz, 4H), 2.65 (t, J= 8 Hz, 4H), 1.98-1.91 (m, 4H).

2B

Methyl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(2-hydroxyphenyl)propanoate.

IV: methyl 2-amino-3-(2-hydroxyphenyl)propanoate hydrochloride.

General method A.

Yield = 16%.

MS ES+: 395.

1H NMR (400 MHz, DMSO-a ₆ ) δ 9.44 (s, 1H), 7.81 (s, 1H), 7.09-7.00 (m, 2H), 6.86 (s, 1H ), 6.79 (d, J=7 Hz, 1H), 6.73-6.70 (m, 1H), 6.27 (d, J=8 Hz, 1H), 4.50-4.45 (m, 1H), 3.60 (s, 3H), 3.04-2.82 (m, 2H), 2.78 (t, J=7 Hz, 4H), 2.67-2.55 ( m, 4H), 1.97-1.90 (m, 4H).

2C

Methyl 3-(3-acetylphenyl)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}propanoate.

IV: methyl 3-(3-acetylphenyl)-2-aminopropanoate.

General method A.

Yield = 71%.

MS ES+: 421.3.

1H NMR (400 MHz, CDCl ₃ ) δ 7.82 (d, J=8 Hz, 1H), 7.66 (s, 1H), 7.35 (t, J=8 Hz, 1H), 7.26 (s, 1H), 7.03 (s, 1H), 5.88 (s, 1H), 4.91-4.82 (m, 2H), 3.75 (s, 3H), 3.27- 3.20 (m, 1H), 3.11-3.04 (m, 1H), 2.88 (t, J=7 Hz, 4H), 2.81-2.72 (m, 2H), 2 .71-2.61 (m, 2H), 2.57 (s, 3H), 2.08-1.97 (m, 4H).

2D

Methyl(2R)-3-(4-cyanophenyl)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}propanoate.

IV: methyl (2R)-2-amino-3-(4-cyanophenyl)propanoate.

General method A.

- 37 044992

Yield = 15%.

MS ES+: 404.2.

1H NMR (400 MHz, DMSO-de) δ 7.83-7.74 (m, 3H), 7.42 (d, J=8 Hz, 2H), 6.87 (s, 1H), 6.42 (d, J=8 Hz, 1H), 4.59-4.53 (m, 1H), 3.66 (s, 3H), 3.19-3.15 (m, 1H), 3.07- 3.01 (m, 1H), 2.78 (t, J=7 Hz, 4H), 2.61-2.54 (m, 4H), 1.94 (quint, J=7 Hz, 4H).

2E

Methyl (2R)-3-(3-cyanophenyl)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}propanoate.

IV: methyl (2R)-2-amino-3-(3-cyanophenyl)propanoate hydrochloride.

General method A.

The product was then purified by crystallization from MeOH to give the title compound as a white solid.

Yield = 5%.

MS ES+: 404.2.

¹ H NMR (400 MHz, DMSO-de) δ 7.81 (s, 1H), 7.72 (d, J=7 Hz, 1H), 7.66 (s, 1H), 7.58-7, 50 (m 2H), 6.87 (s, 1H), 6.42 (d, J=8 Hz, 1H), 4.58-4.52 (m, 1H), 3.67 (s, 3H) , 3.17-3.14 (m, 1H), 3.04-2.99 (m, 1H), 2.78 (t, J=7 Hz, 4H), 2.60 (t, J=7 Hz, 4H), 1.98-1.90 (m, 4H).

2F

Methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(pyridin-2-yl)propanoate.

IV: methyl (2R)-2-amino-3-(pyridin-2-yl)propanoate hydrochloride.

General method A.

Yield = 23%.

MS ES+: 380.

1H NMR (400 MHz, DMSO-de) δ 8.48-8.46 (m, 1H), 7.89 (s, 1H), 7.73-7.71 (m, 1H), 7.30- 7.22 (m, 2H), 6.87 (s, 1H), 6.45 (d, J=8 Hz, 1H), 4.69-4.62 (m, 1H), 3.33 (s , 3H), 3.21-3.13 (m, 2H), 2.78 (t, J=7 Hz, 4H), 2.662.56 (m, 4H), 1.97-1.89 (m, 4H).

2G

Methyl 2-{[(1,2,3,5,6,7-hexαhydro-s-indacen-4-yl)carbamoyl]amino}-3-[3-(hydroxymethyl)phenyl]propanoate.

In a hermetically sealed test tube in anhydrous dioxane in an Ar atmosphere at room temperature, (tributylstannyl)methanol (52 mg, 0.164 mmol, 1.5 eq.), tetrakis(triphenylphosphine) palladium(0) (6.3 mg, 0.005 mmol, 0 .05 eq.) and methyl 3-(3-bromophenyl)-2{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}propanoate (50 mg, 0.109 mmol). The reaction mixture was stirred at 80° C. for 18 hours. After cooling to room temperature, the reaction mixture was concentrated and purified by flash chromatography (DCM/MeOH) to obtain the desired product as a yellow solid.

Yield = 45%.

- 38 044992

MS ES ⁺ : 409.2.

1H NMR (400 MHz, DMSO-d ₆ ) δ 7.86 (s, 1H), 7.26 (t, J=8 Hz, 1H), 7.19 (d, J=8 Hz, 1H), 7 .13 (s, 1H), 7.05 (d, J=7 Hz, 1H), 6.87 (s, 1H), 6.33 (d, J=8 Hz, 1H), 5.16 (t , J=5 Hz, 1H), 4.52-4.43 (m, 3H), 3.64 (s, 3H), 3.06-3.02 (m, 1H), 2.98-2, 92 (m, 1H), 2.79 (t, J=7 Hz, 4H), 2.64 (t, J=7 Hz, 4H), 2.00-1.89 (m, 4H).

2H

Methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(pyridin-3-yl)propanoate.

IV: methyl (2R)-2-amino-3-(pyridin-3-yl)propanoate hydrochloride.

General method A.

The product was then purified using preparative HPLC.

Yield = 6%.

MS ES ⁺ : 380.3.

1H NMR (400 MHz, DMSO) δ 8.46-8.44 (m, 1H), 8.41 (d, J=2 Hz, 1H), 7.85 (s, 1H), 7.63-7 .61 (m, 1H), 7.35-7.32 (m, 1H), 6.87 (s, 1H), 6.45 (d, J=8 Hz, 1H), 4.55-4, 49 (m, 1H), 3.66 (s, 3H), 3.13-3.08 (m, 1H), 3.01 2.95 (m, 1H), 2.78 (t, J=7 Hz, 4H), 2.61 (t, J=7 Hz, 4H), 1.98-1.90 (m, 4H).

2I

Methyl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-[3-(1-methyl-1H-imidazol-5yl)phenyl ]propanoate

In a hermetically sealed test tube in anhydrous dioxane in an Ar atmosphere at room temperature, 1-methyl-5-(tributylstannyl)imidazole (61 mg, 0.164 mmol, 1.5 equiv.), tetrakis(triphenylphosphine)palladium(0) (6.3 mg, 0.005 mmol, 0.05 eq.) and methyl 3-(3-bromophenyl)-2{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino }propanoate (50 mg, 0.109 mmol). The reaction mixture was stirred at 80° C. for 18 hours. After cooling to room temperature, the reaction mixture was concentrated and purified by preparative HPLC to obtain the desired product as a yellow gum.

Yield = 10%.

MS ES ⁺ : 459.4.

1H NMR (400 MHz, DMSO-d6) δ 9.05 (s, 1H), 7.84-7.73 (m, 2H), 7.52-7.45 (m, 3H), 7.41- 7.34 (m, 1H), 6.87 (s, 1H), 6.46 (d, J=8 Hz, 1H), 4.61-4.54 (m, 1H), 3.83 (s , 3H), 3.67 (s, 3H), 3.19-3.14 (m, 1H), 3.07-3.01 (m, 1H), 2.77 (t, J=7 Hz, 4H), 2.60-2.55 (m, 4H), 1.98-1.89 (m, 4H).

2J

Methyl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-[3-(1H-pyrazol-5yl)phenyl]propanoate.

- 39 044992

IV: methyl 2-amino-3-[3-(1H-pyrazol-5-yl)phenyl]propanoate hydrochloride.

General method A.

Yield = 18%.

MS ES+: 445.3.

1H NMR (400 MHz, DMSO- _de ) δ 12.86 (s, 1H), 7.85 (s, 1H), 7.78 (s, 1H), 7.73-7.58 (m, 2H ), 7.33 (t, J=7 Hz, 1H), 7.19-7.06 (m, 1H), 6.85 (s, 1H), 6.67 (s, 1H), 6.37 (d, J=8 Hz, 1H), 4.58-4.49 (m, 1H), 3.66 (s, 3H), 3.13-3.08 (m, 1H), 3.03- 2.97 (m, 1H), 2.77 (t, J=7 Hz, 4H), 2.61 (t, J=7 Hz, 4H), 1.96-1.86 (m, 4H).

2K

Methyl(2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(3-hydroxyphenyl)propanoate.

OH OH

IV: methyl (2K)-2-amino-3-(3-hydroxyphenyl)propanoate hydrochloride.

General method A.

Yield = 50%.

MS ES+: 395.1.

1H NMR (400 MHz, DMSO-de) δ 9.33 (s, 1H), 7.89 (s, 1H), 7.08 (t, J=8 Hz, 1H), 6.87 (s, 1H ), 6.66-6.61 (m, 1H), 6.61-6.56 (m, 2H), 6.30 (d, J=8 Hz, 1H), 4.48-4.43 ( m, 1H), 3.64 (s, 3H), 2.97-2.93 (m, 1H), 2.90-2.85 (m, 1H), 2.79 (t, J=7 Hz , 4H), 2.68-2.60 (m, 4H), 1.98-191 (m, 4H).

2L

Methyl (2H)-2-{[(1,2,3,5,6,7-hexahydro-8-indacen-4-yl)carbamoyl]amino}-3-[3-(1H-pyrazol-3yl)phenyl ]propanoate

IV: methyl (2R)-2-amino-3-[3-(1H-pyrazol-5-yl)phenyl]propanoate hydrochloride.

General method A.

The product was then purified by flash chromatography to give the title compound.

Yield = 32%.

MS ES+: 445.

1H NMR (400 MHz, DMSO- _de ) δ 12.86 (s, 1H), 7.88-7.50 (m, 4H), 7.41-7.28 (m, 1H), 7.19 -7.06 (m, 1H), 6.85 (s, 1H), 6.67 (s, 1H), 6.37 (d, J=8 Hz, 1H), 4.61-4.47 ( m, 1H), 3.66 (s, 3H), 3.13-3.09 (m, 1H), 3.03-2.98 (m, 1H), 2.77 (t, J=7 Hz , 4H), 2.61 (t, J=7 Hz, 4H), 1.95-1.87 (m, 4H).

- 40 044992

2M

Methyl(2R)-2-{[(1,2,3,5,6,7-hexahydro-s-inαcen-4-yl)carbamoyl]amino}-3-[3-(hydroxymethyl)phenyl]propanoate.

In a hermetically sealed test tube, (tributylstannyl)methanol (153 mg, 0.44 mmol, 1 equiv.), tetrakis(triphenylphosphine)palladium(0) (25.2 mg, 0.005 mmol) and methyl (2R)-3-(3-bromophenyl)-2-{[(1,2,3,5,6,7hexahydro-s-indacen-4-yl)carbamoyl]amino}propanoate (200 mg, 0.44 mmol). The reaction mixture was stirred at 80° C. for 18 hours. After cooling to room temperature, the reaction mixture was concentrated and purified by flash chromatography (DCM/MeOH) to obtain the title compound as a yellow solid.

Yield = 10%.

MS ES+: 409.

Ή NMR (400 MHz, DMSO-Y6) δ 7.86 (s, 1H), 7.26 (t, J=8 Hz, 1H), 7.19 (d, J=8 Hz, 1H), 7, 13 (s, 1H), 7.05 (d, J=8 Hz, 1H), 6.87 (s, 1H), 6.34 (d, J=8 Hz, 1H), 5.16 (t, J=6 Hz, 1H), 4.52-4.43 (m, 3H), 3.64 (s, 3H), 3.06-3.02 (m, 1H), 2.98-2.93 (m, 1H), 2.79 (t, J=7 Hz, 4H), 2.64 (t, J=7 Hz, 4H), 1.98-1.91 (m, 4H).

2N

Methyl(2R)-3-amino-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}propαnoαt.

IV: methyl (2R)-3-{[(tert-butoxy)carbonyl]amino}-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4yl)carbamoyl] amino} propanoate

General method C.

Yield = 82%.

MS ES+: 318.2.

¹ H NMR (400 MHz, DMSO-d6) δ 8.22 (s, 1H), 8.14 (s, 3H), 6.90 (s, 1H), 6.83 (d, J=8 Hz, 1H), 4.54-4.49 (m, 1H), 3.70 (s, 3H), 3.25-3.21 (m, 1H), 3.16-3.03 (m, 1H) , 2.80 (t, J=7 Hz, 4H), 2.72 (t, J=7 Hz, 4H), 2.00-1.93 (m, 4H).

2O

Methyl(2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-4-(methylsulfanyl)butanoate.

IV: D-methionine methyl ester hydrochloride General method A.

Yield = 72%.

MS ES+: 363.2.

¹ H NMR (400 MHz, DMSO-a ₆ ) δ 7.78 (s, 1H), 6.88 (s, 1H), 6.52 (d, J=8 Hz, 1H), 4.35-4 .30 (m, 1H), 3.66 (s, 3H), 2.79 (t, J=7 Hz, 4H), 2.68 (t, J=7 Hz, 4H), 2.06 (s , 3H), 2.00-1.85 (m, 6H).

2P

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Methyl(2R)-3-(3-acetamidophenyl)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}propanoate.

IV: methyl (2R)-2-amino-3-(3-acetamidophenyl)propanoate hydrochloride.

General method A.

The title compound was then purified by FCC to obtain the title compound as a white solid.

Yield = 11%.

MS ES+: 436.4.

¹ H NMR (400 MHz, DMSO-d^ δ 7.91 (s, 1H), 6.93 (t, J=8 Hz, 1H), 6.87 (s, 1H), 6.47-6, 40 (m, 1H), 6.406.35 (m, 1H), 6.32-6.24 (m, 2H), 5.00 (s, 1H), 4.46-4.34 (m, 1H) , 3.64 (s, 2H), 2.91-2.75 (m, 6H), 2.67-2.63 (m, 4H), 1.98-1.91 (m, 4H).

2Q

Methyl 2-{[(1,2,3,5,6,7-hexahydro-8-indacen-4-yl)carbamoyl]amino}-3-(1-methyl-1H-pyrazol4-yl)propanoate.

IV: Methyl 2-amino-3-(1-methyl-1H-pyrazol-4-yl)propanoate dihydrochloride.

General method A.

The title compound was then purified by FCC to obtain the title compound as a white solid.

Yield = 5%.

MS ES+: 383.3.

Ή NMR (400 MHz, DMSO-a ₆ ) δ 7.91 (s, 1H), 7.45 (s, 1H), 7.22 (d, J=1 Hz, 1H), 6.88 (s, 1H), 6.30 (d, J=8 Hz, 1H), 4.45-4.35 (m, 1H), 3.79 (s, 3H), 3.66 (s, 3H), 2, 91-2.82 (m, 2H), 2.79 (t, J=7 Hz, 4H), 2.67 (t, J=7 Hz, 4H), 1.99-1.92 (m, 4H ).

2R.

Methyl(2R)-2-{[(1,2,3,5,6,7-hexαhydro-s-indacen-4-yl)carbamoyl]amino}-3-[3-(2-oxopyrrolidin1-yl)phenyl ]propanoate

- 42 044992

IV: methyl (2R)-2-amino-3-{3-[(2-oxocyclopentyl)amino]phenyl}propanoate.

General method A.

The title compound was then purified by FCC to obtain the title compound as a white solid.

Yield = 38%.

MS ES+: 462.8.

¹ H NMR (400 MHz, DMSO-b ₆ ) δ 7.85 (s, 1H), 7.62-7.54 (m, 1H), 7.46 (s, 1H), 7.30 (t, J=8 Hz, 1H), 6.95 (d, J=8 Hz, 1H), 6.87 (s, 1H), 6.33 (d, J=8 Hz, 1H), 4.53-4 .48 (m, 1H), 3.89-3.76 (m, 2H), 3.66 (s, 3H), 3.10-2.90 (m, 2H), 2.78 (t, J =7 Hz, 4H), 2.62 (t, J=7 Hz, 4H), 2.10-2.02 (m, 2H), 1.97-1.90 (m, 4H).

2S

1,5-dimethyl(2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}pentanedioate.

IV: 1,5-dimethyl (2R)-2-aMUHoneHTaHguoama hydrochloride.

General method A.

Yield = 34%.

MS ES+: 375.2.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 7.80 (s, 1H), 6.88 (s, 1H), 6.49 (d, J=8 Hz, 1H), 4.28-4 .22 (m, 1H), 3.65 (s, 3H), 3.60 (s, 3H), 2.80 (t, J=7 Hz, 4H), 2.68 (t, J=7 Hz , 4H), 2.45-2.35 (m, 2H), 2.05-1.92 (m, 5H), 1.91-1.80 (m, 1H).

2T

Ethyl 2-[({1H,2H,3H,6H,7H,8H,9H-cyclopenta[a]naphthalene-5-yl}carbamoyl)amino]acetate.

1H,2H,3H,6H,7H,8H,9H-cyclopenta[a]naphthalene-5-amine (20 mg, 0.107 mmol) was dissolved in ACN (1 ml). A solution of ethyl 2-isocyanatoacetate (17 mg, 1.2 eq., 0.128 mmol) in ACN (1 mL) was added and the reaction mixture was stirred at room temperature overnight. The resulting precipitate was filtered, washed with ACN and dried under reduced pressure to obtain the title compound as a white solid.

Yield = 97%.

MS ES+: 317.

Ή NMR (400 MHz, DMSO-d ₆ ) δ 7.69 (s, 1H), 7.34 (s, 1H), 6.68 (t, J=6 Hz, 1H), 4.14-4, 08 (m, 2H), 3.84 (d, J=6 Hz, 2H), 2.78 (t, J=8 Hz, 2H), 2.67 (t, J=7 Hz, 2H), 2 .55 (t, J=5 Hz, 2H), 2.52-2.46 (m, 6H), 2.01-1.94 (m, 2H), 1.76-1.66 (m, 4H ), 1.21 (t, J=7 Hz, 3H).

2U

Methyl(2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-ul)carbamoyl]amino}-3-{3-[(1H-pyrazol-3yl) amino]phenyl}propanoate.

IV: (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-{3-[(1H-pyrazol-3yl )amino]phenyl}propanoic acid.

General method B.

The product was then purified by preparative HPLC to obtain the desired product as a white solid.

- 43 044992

Yield = 38%.

MS ES ⁺ : 460.

1H NMR (400 MHz, DMSO-b ₆ ) δ 8.40 (s, 1H), 7.90 (s, 1H), 7.57 (d, J=2 Hz, 1H), 7.18-7, 13 (m, 2H), 7.127.07 (m, 1H), 6.86 (s, 1H), 6.53 (d, J=7 Hz, 1H), 6.26 (d, J=8 Hz, 1H), 5.86 (d, J=2 Hz, 1H), 4.47 (m, 1H), 3.65 (s, 3H), 2.97-2.86 (m, 2H), 2, 78 (t, J=7 Hz, 4H), 2.66-2.62 (m, 4H), 1.97-1.90 (m, 4H).

2V (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl] amino }-3-[3-(1H-pyrazole-3 - yl)phenyl] propanoic acid.

(2R)-2-Amino-3-[3-(1H-pyrazol-3-yl)phenyl]propanoic acid dihydrochloride (180 mg, 0.59 mmol) was dissolved IN 1 M NaOH (0.7 ml, 1.20 mmol ) and cooled to a temperature of 0°C. The resulting solution was then treated dropwise with a solution of intermediate A (120 mg, 0.60 mmol) in acetone (1.4 ml). After stirring at room temperature for 24 hours, a further portion of Intermediate A (120 mg, 0.60 mmol) in acetone (1.4 ml) was added. The reaction mixture was stirred at room temperature for an additional 24 hours. The reaction mixture was filtered and the collected solid triturated in acetone. It was then purified by preparative HPLC to obtain the title compound as a white powder.

Yield = 7%.

MS ES ⁺ : 431.1.

1H NMR (400 MHz, DMSO-a ₆ ) δ 12.92 (s, 2H), 7.87 (s, 1H), 7.70 (s, 1H), 7.67-7.61 (m, 2H ), 7.32 (t, J=8 Hz, 1H), 7.14 (d, J=8 Hz, 1H), 6.84 (s, 1H), 6.64 (d, J=2 Hz, 1H), 6.25 (d, J=8 Hz, 1H), 4.46-4.41 (m, 1H), 3.173.12 (m, 1H), 3.01-2.96 (m, 1H ), 2.77 (t, J=7 Hz, 4H), 2.62 (t, J=7 Hz, 4H), 1.94-1.87 (m, 4H).

2X

Ethyl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}acetate.

2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}acetic acid (80 mg, 0.3 mmol) was suspended in dioxane (1 ml) and then treated with 1,1'-carbonyldiimidazole (61 mg, 0.37 mmol). After stirring at room temperature for 15 minutes, ethanol (2 ml, 33.5 mmol) was added and the reaction mixture was refluxed for 4 hours. The reaction mixture was evaporated, triturated in water, filtered and recrystallized from boiling ethanol to give the above The title compound is in the form of a white powder.

Yield = 22%.

MS ES ⁺ : 303.1.

1H NMR (400 MHz, DMSO) δ 7.92 (s, 1H), 6.89 (s, 1H), 6.33 (t, J=6 Hz, 1H), 4.13-4.07 (m , 2H), 3.81 (d, J=6 Hz, 2H), 2.80 (t, J=7 Hz, 4H), 2.70 (t, J=7 Hz, 4H), 2.09- 1.84 (m, 4H), 1.21 (t, J=7 Hz, 3H).

2W

1,4-dimethyl(2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}butanedioate.

- 44 044992

IV: 1,4-dimethyl (2R)-2-aminobutanedioate hydrochloride.

General method A.

Yield = 46.7%.

MS ES+: 361.0.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.02 (s, 1H), 6.88 (s, 1H), 6.57 (d, J=8.5 Hz, 1H), 4.64 -4.54 (m, 1H), 3.65 (s, 3H), 3.63 (s, 3H), 2.87-2.74 (m, 6H), 2.67 (t, J=7 ,3 Hz, 4H), 2.02-1.88 (m, 4H).

2Y

Methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-phenylpropanoate.

IV: methyl (2R)-2-amino-3-phenylpropanoate hydrochloride.

General method A.

Yield = 44%.

MS ES+: 379.1.

¹ H NMR (400 MHz, DMSO) δ 7.85 (s, 1H), 7.49-7.05 (m, 5H), 6.87 (s, 1H), 6.33 (d, J=8 Hz, 1H), 4.534.44 (m, 1H), 3.64 (s, 3H), 3.11-3.01 (m, 1H), 3.0-2.92 (m, 1H), 2 .79 (t, J=7 Hz, 4H), 2.63 (t, J=7 Hz, 4H), 2.031.87 (m, 4H).

2Z

Methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(4-hydroxyphenyl)propanoate.

IV: methyl (2R)-2-amino-3-(4-hydroxyphenyl)propanoate hydrochloride.

General method A.

Yield = 33%.

MS ES+: 395.3.

¹ H NMR (400 MHz, DMSO) δ 9.25 (s, 1H), 7.86 (s, 1H), 6.96 (d, J=8 Hz, 2H), 6.87 (s, 1H) , 6.68 (d, J=8 Hz, 2H), 6.24 (d, J=8 Hz, 1H), 4.44-4.37 (m, 1H), 2.96-2.83 ( m, 2H), 2.79 (t, J=7 Hz, 4H), 2.64 (t, J=7 Hz, 4H), 2.0-1.90 (m, 4H).

2AA.

Propan-2-yl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}acetate.

IV: propane hydrochloride -2-yl 2-aminoacetate.

General method A.

Yield = 58%.

MS ES+: 317.2.

¹ H NMR (400 MHz, DMSO) δ 7.91 (s, 1H), 6.89 (s, 1H), 6.32 (t, J=6 Hz, 1H), 4.97-4.88 ( m, 1H), 3.78 (d, J=6 Hz, 2H), 2.80 (t, J=7 Hz, 4H), 2.71 (t, J=7 Hz, 4H), 2.0 -1.92 (m, 4H), 1.22 (d, J=6 Hz, 6H).

2BB

Methyl (2R)-3-carbamoyl-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}propanoate.

- 45 044992

IV: methyl (2R)-2-amino-3-carbamoylpropanoate hydrochloride.

General method A.

Yield = 50%.

MS ES+: 346.1.

^1H NMR (400 MHz, DMSO) δ 8.07 (s, 1H), 7.46 (s, 1H), 6.96 (s, 1H), 6.87 (s, 1H), 6.44 ( d, J=8 Hz, 1H), 4.50 (s, 1H), 3.62 (s, 3H), 2.79 (t, J=7 Hz, 4H), 2.74-2.64 ( m, 5H), 2.56 (d, J=4 Hz, 1H), 2.04-1.89 (m, 4H).

2SS

Methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(thiophen-2-yl)propanoate.

IV: methyl (2R)-2-amino-3-(thiophen-2-yl)propanoate hydrochloride.

General method A.

Yield = 81%.

MS ES+: 385.1.

^1H NMR (400 MHz, DMSO) δ 7.99 (s, 1H), 7.41-7.38 (m, 1H), 7.0-6.97 (m, 1H), 6.88 (s , 2H), 6.43 (d, J=8 Hz, 1H), 4.55-4.48 (m, 1H), 3.67 (s, 3H), 3.31-3.20 (m, 2H), 2.80 (t, J=7 Hz, 4H), 2.68 (t, J=7 Hz, 4H), 2.00-1.92 (m, 4H).

2DD

Methyl 2-{[(1,2,3,5,6,7-hexahydro-s-ingacen-4-yl)carbamoyl]amino}-3-(1H-imidazol-1-yl)propanoate.

IV: methyl 2-amino-3-(1H-imidazol-1-yl)propanoate hydrochloride.

General method A.

Yield = 64%.

MS ES+: 369.2.

^1H NMR (400 MHz, DMSO) δ 7.97 (s, 1H), 7.53 (s, 1H), 7.07 (s, 1H), 6.90 (d, J=4 Hz, 2H) , 6.51 (d, J=8 Hz, 1H), 4.65-4.55 (m, 1H), 4.46-4.27 (m, 2H), 3.69 (s, 3H), 2.80 (t, J=7 Hz, 4H), 2.67 (t, J=7 Hz, 4H), 2.00-1.93 (m, 4H).

2EE

Methyl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}acetate.

IV: intermediate A and methyl 2-aminoacetate hydrochloride.

General method A.

Yield = 68%.

MS ES+: 289.0.

- 46 044992 ¹ H NMR (400 MHz, DMSO) δ 7.92 (s, 1H), 6.89 (s, 1H), 6.34 (t, J=6 Hz, 1H), 3.84 (d , J=6 Hz, 2H), 3.64 (s, 3H), 2.80 (t, J=7 Hz, 4H), 2.70 (t, J=7 Hz, 4H), 2.09- 1.84 (m, 4H).

2FF (2S)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-phenylpropanoic acid.

IV: (2S)-2-amino-3-phenylpropanoic acid.

General method A.

Yield = 62%.

MS ES+: 365.1;

¹ H NMR (400 MHz, DMSO-a ₆ ) δ 12.76 (s, 1H), 7.88 (s, 1H), 7.36-7.26 (m, J=7 Hz, 2H), 7 ,27-7.17 (m, 3H), 6.86 (s, 1H), 6.23 (d, J=8 Hz, 1H), 4.49-4.39 (m, 1H), 3, 13-3.05 (m, 1H), 2.98-2.90 (m, 1H), 2.79 (t, J=7 Hz, 4H), 2.65 (t, J=7 Hz, 4H ), 2.00-1.90 (m, 4H).

2GG

Methyl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-phenylpropanoate.

Mixture: methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-phenylpropanoate and methyl (2S)-2- {[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl] amino} -3-phenylpropanoate.

2NN

Methyl(2R)-3-(3-aminophenyl)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}propanoate.

IV: intermediate A and methyl (2R)-2-amino-3-(3-acetamidophenyl)propanoate.

General method A.

Yield = 10%.

MS ES+: 394.7.

¹ H NMR (400 MHz, DMSO-b ₆ ) δ 7.91 (s, 1H), 6.93 (t, J=8 Hz, 1H), 6.87 (s, 1H), 6.47-6 .40 (m, 1H), 6.406.35 (m, 1H), 6.34-6.23 (m, 2H), 5.00 (s, 1H), 4.46-4.34 (m, 1H ), 3.64 (s, 2H), 2.91-2.75 (m, 6H), 2.65 (t, J=7.8 Hz, 4H), 2.01-1.89 (m, 4H).

2II

Methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-methoxypropanoate.

IV: methyl (2R)-2-amino-3-methoxypropanoate hydrochloride.

General method A.

Yield = 75%.

MS ES+: 333.1.

Ή NMR (400 MHz, DMSO-b ₆ ) δ 7.99 (s, 1H), 6.87 (s, 1H), 6.51 (d, J=9 Hz, 1H), 4.45-4, 37 (m, 1H), 3.773.70 (m, 1H), 3.66 (s, 3H), 3.61-3.52 (m, 1H), 3.28 (s, 3H), 2.79 (t, J=7 Hz, 4H), 2.68 (t, J=7 Hz, 4H), 2.03-1.89 (m, 4H).

2JJ

Methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-hydroxypropanoate.

- 47 044992

IV: methyl hydrochloride (2R)-2-aMUHO-3-rugpoKCunponaHoama.

General method A.

The compound was then purified by trituration with minimal DMSO. The resulting solid was filtered, washed successively with ACN and Et ₂ O and dried in vacuo to obtain the title compound as a white powder.

Yield = 68%.

MS ES+: 319.1.

¹ H NMR (400 MHz, DMSO-d6) δ 8.04 (s, 1H), 6.88 (s, 1H), 6.46 (d, J=8 Hz, 1H), 5.17 (t, J=4 Hz, 2H), 4.36-4.19 (m, 1H), 3.90-3.73 (m, 1H), 3.66 (s, 3H), 2.80 (t, J =6 Hz, 4H), 2.75-2.63 (m, 4H), 2.05-1.88 (m, 4H).

2KK

Ethyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-[3-(1H-pyrazol-4yl)phenyl ]propanoate

IV: ethyl (2R)-2-amino-3-[3-(1H-pyrazol-3-yl)phenyl]propanoate hydrochloride.

General method A.

The compound was then purified by preparative TLC (hexane:ethyl acetate 4:1) to give the title compound as a yellow solid.

Yield = 1%.

MS ES+: 459.2.

Ή NMR (400 MHz, DMSO-d ₆ ) δ 12.93 (broad s, 1H), 8.24 (broad s, 1H), 7.76-7.59 (m, 3H), 7.33 (t , J=8 Hz, 1H), 7.14 (d, J=8 Hz, 1H), 6.84 (s, 1H), 6.80 (wide s, 1H), 6.67 (d, J= 2 Hz, 1H), 4.52-4.41 (m, 1H), 4.14-4.03 (m, 2H), 3.12-2.96 (m, 2H), 2.77 (t , J=7 Hz, 4H), 2.62 (t, J=7 Hz, 4H), 1.98-1.90 (m, 4H), 1.16 (t, J=7 Hz, 3H).

2LL (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(pyridin-3-yl)propanoic acid.

To a suspension of (2R)-2-amino-3-(pyridin-3-yl)propanoic acid (200 mg, 1.2 mmol, 1 eq.) in acetone/H ₂ O (1:1, 8 ml) was added Et ₃ N (252 µl, 1.8 mmol, 1.5 eq) and the mixture was stirred for 5 min. A solution of intermediate A (264 mg, 1.3 mmol, 1.1 eq) in THF (2 ml) was added and the reaction mixture was stirred overnight at room temperature. The volume of the mixture was reduced by half in vacuo and the resulting white precipitate was filtered off, washed successively with water, ACN and Et ₂ O and dried in vacuo to obtain the title compound as a white powder.

Yield = 68%.

MS ES+: 366.3.

Ή NMR (400 MHz, DMSO-b ₆ ) δ 12.91 (s, 1H), 8.44 (d, J=4 Hz, 1H), 8.41 (s, 1H), 7.86 (s, 1H), 7.61 (d, J=7 Hz, 1H), 7.37-7.29 (m, 1H), 6.86 (s, 1H), 6.32 (d, J=8 Hz, 1H), 4.50-4.38 (m, 1H), 3.18-3.06 (m, 1H), 3.012.91 (m, 1H), 2.78 (t, J=7 Hz, 4H ), 2.63 (t, J=7 Hz, 4H), 1.98-1.90 (m, 4H).

2MM

2-Methoxyethyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(pyridin-3yl)propanoate.

- 48 044992

IV: 2-methoxyethyl (2R)-2-amino-3-(pyridin-3-yl)propanoate hydrochloride.

General method A.

Yield = 28%.

MS ES+: 424.5.

1H NMR (400 MHz, DMSO-d6) δ 8.47-8.44 (m, 1H), 8.42 (d, J=2 Hz, 1H), 7.92 (s, 1H), 7.68 -7.59 (m, 1H), 7.38-7.30 (m, 1H), 6.87 (s, 1H), 6.50 (d, J=8 Hz, 1H), 4.58- 4.49 (m, 1H), 4.26-4.14 (m, 2H), 3.59-3.48 (m, 2H), 3.28 (s, 3H), 3.14-3. 06 (m, 1H), 3.05-2.96 (m, 1H), 2.79 (t, J=7 Hz, 4H), 2.62 (t, J=7 Hz, 4H), 1, 98-1.90 (m, 4H).

2NN

Cyclobutyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(pyridin-3yl)propanoate.

IV: cyclobutyl (2R)-2-amino-3-(pyridin-3-yl)propanoate.

General method A.

The crude product was then purified by preparative HPLC (HCOOH buffer) to obtain the title compound as a white solid.

Yield = 2%.

MS ES+: 420.4.

1H NMR (400 MHz, DMSO-d6) δ 8.51-8.35 (m, 2H), 7.91 (s, 1H), 7.68-7.59 (m, 1H), 7.38- 7.29 (m, 1H), 6.87 (s, 1H), 6.49 (d, J=8 Hz, 1H), 4.96-4.85 (m, 1H), 4.50-4 .39 (m, 1H), 3.12-3.04 (m, 1H), 3.04-2.95 (m, 1H), 2.78 (t, J=7 Hz, 4H), 2, 63 (t, J=7 Hz, 4H), 2.30-2.20 (m, 2H), 1.99-1.89 (m, 4H), 1.80-1.68 (m, 1H) , 1.67-1.52 (m, 1H).

2OO

Cyclopropylmethyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(pyridin3-yl)propanoate.

IV: cyclopropylmethyl (2R)-2-amino-3-(pyridin-3-yl)propanoate.

General method A.

The product was purified by preparative HPLC (HCOOH buffer) to obtain the title compound as a white solid.

Yield = 8%.

MS ES+: 420.4.

1H NMR (400 MHz, DMSO-d ₆ ) δ 8.48-8.40 (m, 2H), 7.93 (s, 1H), 7.64 (d, J=8 Hz, 1H), 7, 38-7.29 (m, 1H), 6.87 (s, 1H), 6.51 (d, J=8 Hz, 1H), 4.56-4.47 (m, 1H), 3.91 (d, J=7 Hz, 2H), 3.13-3.05 (m, 1H), 3.05-2.97 (m, 1H), 2.78 (t, J=7 Hz, 4H) , 2.63 (t, J=7 Hz, 4H), 2.02-1.85 (m, 4H), 1.13-0.99 (m, 1H), 0.56-0.47 (m , 2H), 0.34-0.21 (m, 2H).

2PP

Cyclopentyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(pyridin-3yl)propanoate.

IV: cyclopentyl (2R)-2-amino-3-(pyridin-3yl)propanoate ditrifluoromethanesulfonic acid salt.

General method A.

The crude product was purified by preparative HPLC (HCOOH buffer) to obtain the title compound as a white solid.

Yield = 9%.

MS ES+: 434.5.

1H NMR (400 MHz, DMSO-d ₆ ) δ 8.45 (d, J=5 Hz, 1H), 8.43-8.38 (m, 1H), 7.85 (s, 1H), 7, 63 (d, J=8 Hz, 1H), 7.40-7.29 (m, 1H), 6.88 (s, 1H), 6.42 (d, J=8 Hz, 1H), 5, 12-5.03 (m, 1H), 4.47-4.37 (m, 1H), 3.11-2.94 (m, 2H), 2.79 (t, J=7 Hz, 4H) , 2.63 (d, J=7 Hz, 4H), 1.95 2.02-1.88 (m, 4H), 1.87-1.72 (m, 2H), 1.67-1, 46 (m, 6H).

2QQ

Methyl (2R)-3-cyano-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}propanoate.

Methyl (2R)-3-carbamoyl-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}propanoate (Example 2BB) (103 mg, 0 .3 mmol, 1 eq.) was suspended in anhydrous DCM (3 ml). Para-tosyl chloride (239 mg, 1.2 mmol, 4.2 eq) was added followed by pyridine (240 µl, 3 mmol, 10 eq) and the reaction mixture was stirred at room temperature under argon for 72 h. the mixture was evaporated and the resulting solid was washed successively with DCM, H ₂ O and Et ₂ O. The crude product was purified by preparative HPLC (formic acid buffer) to give the title compound.

Yield = 13%.

MS ES+: 350.3 [M+Na]+.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.09 (s, 1H), 6.90 (s, 1H), 6.82 (d, J=8 Hz, 1H), 4.61-4 .53 (m, 1H), 3.69 (s, 3H), 3.11-2.95 (m, 2H), 2.80 (t, J=7 Hz, 4H), 2.70 (t, J=7 Hz, 4H), 2.91-1.93 (m, J=7 Hz, 4H).

2RR

Ethyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(pyridin-3-yl)propanoate.

IV: (2R)-2-amino-3-(pyridin-3-yl)propanoic acid hydrochloride.

General method A.

Yield = 69% MS ES+: 394.5.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.45 (d, J=3 Hz, 2H), 8.42 (s, 1H), 7.88 (s, 1H), 7.63 (d , J=7 Hz, 1H), 7.37-7.28 (m, 1H), 6.87 (s, 1H), 6.46 (d, J=8 Hz, 1H), 4.54-4 .41 (m, 1H), 4.15-4.07 (m, 1H), 3.14-2.94 (m, 2H), 2.79 (t, J=7 Hz, 4H), 2, 62 (t, J=7 Hz, 4H), 1.98-1.90 (m, J=7 Hz, 4H), 1.17 (t, J=7 Hz, 3H).

2SS (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-phenylpropanoic acid.

Methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-phenylpropanoate [intermediate 2Y] (31 mg, 0 .08 mmol) was dissolved in MeOH (2 ml) and water (2 ml), then treated with LiOH-H ₂ O (6 mg, 0.14 mmol). The reaction mixture was stirred at room temperature for 18 hours.

2TT

Methyl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(4-methyl-1H-pyrazol1 -yl)propanoate.

IV: methyl 2-amino-3-(4-methyl-1H-pyrazol-1-yl)propanoate hydrochloride. General method A.

- 50 044992

Yield = 13%.

MS ES+: 383.3.

¹ H NMR (400 MHz, DMSO-d ₆ ) δ 8.05 (s, 1H), 7.38 (t, J=1 Hz, 1H), 7.26 (t, J=1 Hz, 1H), 6.89 (s, 1H),

6.36 (d, J=8 Hz, 1H), 4.67-4.58 (m, 1H), 4.50-4.35 (m, 2H), 3.66 (s, 3H), 2 .80 (t, J=7 Hz, 4H), 2.66 (t, J=7 Hz,

4H), 2.03-1.89 (m, 7H).

2UU

Ethyl (2R)-2-({[2,6-bis(propan-2-yl)phenyl]carbamoyl}amino)-3-(pyridin-3-yl)propanoate.

Methyl (2R)-2-amino-3-(3-cyanophenyl)propanoate hydrochloride (50 mg, 0.217 mol), 4-N,N-dimethylaminopyridine (ca. 2 mg) and MeCN (1 ml) were placed in the vessel. A solution of 2,6diisopropylphenyl isocyanate (43 mg, 0.217 mmol) in MeCN (1 mL) was added, followed by triethylamine (0.076 mL, 0.54 mmol). The vessel was hermetically sealed and stirred at room temperature for 18 hours. The resulting solution was diluted with DCM, washed with water, dried over Na ₂ SO ₄ and evaporated. The crude product was purified using FCC (silica gel, 0-100% EtOAc in hexane) to obtain the desired product as a white solid.

Yield = 48%.

MS ES+: 398.3.

1H NMR (400 MHz, DMSO-a ₆ ) δ 8.45 (d, J=5 Hz, 2H), 7.64 (d, J=7 Hz, 1H), 7.57 (s, 1H), 7 ,40-7.30 (m, 1H), 7.19 (t, J=8 Hz, 1H), 7.10 (s, 1H), 7.08 (s, 1H), 6.55 (d, J=7 Hz, 1H), 4.55-4.45 (m, 1H), 4.09 (kV, J=7 Hz, 2H), 3.19-3.05 (m, 2H), 3, 05-2.95 (m, 2H), 1.17 (t, J=7 Hz, 3H), 1.09 (d, J=6 Hz, 12H).

2VV

Ethyl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(pyrimidin-2-yl)propanoate.

IV: ethyl 2-amino-3-(pyrimidin-2-yl)propanoate dihydrochloride.

General method A.

Yield = 43%.

MS ES+: 395.5.

1H NMR (400 MHz, DMSO-b ₆ ) δ 9.77 (s, 1H), 8.74 (d, J=5 Hz, 2H), 7.94 (s, 1H), 7.39 (t, J=5 Hz, 1H), 6.87 (s, 1H), 6.51 (d, J=9 Hz, 1H), 4.83-4.72 (m, 1H), 4.10-4, 00 (m, 3H), 3.13-3.02 (m, 6H), 2.78 (t, J=l Hz, 4H), 2.66-2.58 (m, 4H), 2.00 -1.86 (m, 4H), 1.19 (t, J=7 Hz, 9H), 1.10 (t, J=7 Hz, 3H). In combination with triethylamine hydrochloride.

2WW

Methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(5-methoxypyridin-3yl)propanoate.

IV: methyl (2R)-2-amino-3-(5-methoxypyridin-3-yl)propanoate dihydrochloride.

General method A.

The product was purified by acid preparative HPLC, then purified by preparative TLC (silica gel, 9:1 DCM/MeOH isocratic).

Yield = 15%.

MS ES+: 410.5.

Ή NMR (300 MHz, CDCl ₃ ) δ 8.19 (s, 1H), 7.91 (s, 1H), 7.05 (s, 1H), 6.96 (s, 1H), 5.85 ( s, 1H), 4.92-4.82 (m, 2H), 3.82 (s, 3H), 3.75 (s, 3H), 3.23-3.15 (m, 1H), 3 .05-2.97 (m, 1H), 2.89 (t, J=8 Hz, 4H), 2.77-2.67 (m,

- 51 044992

4H), 2.10-2.00 (m, 4H).

2XX

Ethyl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(3-methyl-1,2,4-oxadiazol-5 yl )propanoate

IV: ethyl 2-amino-3-(3-methyl-1,2,4-oxadiazol-5-yl)propanoate hydrochloride.

General method A.

The crude product was purified by acid preparative HPLC to obtain the desired product.

Yield = 23%.

MS ES+: 399.5.

1H NMR (400 MHz, DMSO-Y6) δ 8.00 (s, 1H), 6.89 (s, 1H), 6.61 (d, J=8 Hz, 1H), 4.75-4.67 (m, 1H), 4.164.07 (m, 2H), 3.46-3.35 (m, 2H), 2.79 (t, J=7 Hz, 4H), 2.65 (t, J= 7 Hz, 4H), 2.32 (s, 3H), 2.01-1.90 (m, 4H), 1.17 (t, J=7 Hz, 3H).

2YY

Methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(pyridazin-3yl)propanoate.

IV: methyl (2R)-2-amino-3-(pyridazin-3-yl)propanoate hydrochloride.

General method A.

The crude product was purified by acid preparative HPLC to obtain the desired product as a white solid.

Yield = 23%.

MS ES+: 381.4.

Ή NMR (400 MHz, DMSO-Y6) δ 9.12 (dd, J=5, 2 Hz, 1H), 7.90 (s, 1H), 7.66-7.58 (m, 2H), 6 .86 (s, 1H), 6.53 (d, J=8 Hz, 1H), 4.76-4.71 (m, 1H), 3.63 (s, 3H), 3.44-3, 32 (m, 2H), 2.79-2.75 (m, 4H), 2.60-2.56 (m, 4H), 1.96-1.89 (m, 4H).

2ZZ

Ethyl 2-{[(1,2,3,5,6,7-hexahydro-8-indacen-4-yl)carbamoyl]amino}-3-(1,2-oxazol-4-yl)propanoate.

O. .SNz

IV: ethyl 2-amino-3-(1,2-oxazol-4-yl)propanoate hydrochloride.

General method A.

The crude product was purified by acid preparative HPLC to obtain the desired product as a white solid.

Yield = 16%.

MS ES+: 384.8.

^1H NMR (400 MHz, DMSO- _d6 ) δ 8.73 (s, 1H), 8.47 (s, 1H), 7.88 (s, 1H), 6.88 (s, 1H), 6 .49 (d, J=8 Hz, 1H), 4.49-4.40 (m, 1H), 4.11 (q, J=8 Hz, 2H), 2.97-2.84 (m, 2H), 2.81-2.78 (m, 4H), 2.68-2.64 (m, 4H), 1.99-1.92 (m, 4H), 1.19 (t, J= 8 Hz, 3H).

2AB

Ethyl 2-{[(1,2,3,5,6,7-hexahydro-8-indacen-4-yl)carbamoyl]amino}-3-(1,2-oxazol-3-yl)propanoate.

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IV: ethyl 2-amino-3-(1,2-oxazol-3-yl)propanoate hydrochloride.

General method A.

Yield = 66%.

MS ES+: 384.

1H NMR (400 MHz, DMSO-66) δ 8.85 (d, J=2 Hz, 1H), 7.95 (s, 1H), 6.88 (s, 1H), 6.51-6.48 (m, 2H), 4.624.53 (m, 1H), 4.17-4.06 (m, 2H), 3.19-3.06 (m, 2H), 2.81-2.78 (m , 4H), 2.68-2.64 (m, 4H), 2.00-1.89 (m, 4H), 1.18 (t, J=7 Hz, 3H).

2AC

Ethyl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(1,3-oxazol-2-yl)propanoate.

IV: ethyl 2-amino-3-(1,3-oxazol-2-yl)propanoate hydrochloride.

General method A.

Yield = 21%.

MS ES+: 384.4.

¹ H NMR (400 MHz, DMSO-b ₆ ) δ 8.05 (s, 1H), 8.03 (s, 1H), 7.15 (s, 1H), 6.88 (s, 1H), 6 .53 (d, J=8 Hz, 1H), 4.69-4.63 (m, 1H), 4.14-4.04 (m, 2H), 3.23 (d, J=6 Hz, 2H), 2.82-2.76 (m, 4H), 2.68-2.63 (m, 4H), 1.99-1.90 (m, 4H), 1.16 (t, J= 7 Hz, 3H).

2AD

Ethyl (2R)-2-({[6-(propan-2-yl)-2,3-dihydro-1H-inden-5-yl]carbamoyl}amino)-3-(pyridin-3yl)propanoate.

A solution of 5-isocyanato-6-(propan-2-yl)-2,3-dihydro-1H-indene (30 mg, 0.15 mmol), ethyl (2R)-2-amino-3-(pyridine-3) hydrochloride -yl)propanoate (34 mg, 0.15 mmol) and DMAP (small end of spatula) in acetonitrile (3 ml) were treated with triethylamine (52 μl, 0.37 mmol). The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was concentrated, diluted with 1M HCl and extracted with DCM. The organic phase was dried over sodium sulfate and evaporated. The resulting solid was suspended in hexane, filtered and washed with Et ₂ O. The crude product was then purified using prep. TLC (silica gel, EtOAc/hexane) to obtain the desired product.

Yield = 22%.

MS ES+: 396.2.

1H NMR (400 MHz, DMSO-b ₆ ) δ 8.52-8.48 (m, 2H), 7.77-7.72 (m, 2H), 7.46-7.40 (m, 1H) , 7.25 (s, 1H), 7.07 (s, 1H), 6.72 (d, J=9 Hz, 1H), 4.58-4.49 (m, 1H), 4.15- 4.06 (m, 2H), 3.15-2.96 (m, 3H), 2.82-2.71 (m, 4H), 2.01-1.91 (m, 2H), 1, 20-1.09 (m, 9H).

2AE

Methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(pyrimidin-5yl)propanoate.

- 53 044992

IV: methyl (2H)-2-amino-3-(pyrimidin-5-yl)propanoate hydrochloride.

General method A.

The crude product was purified by acid preparative HPLC to obtain the desired product as a white solid.

Yield = 85%.

MS ES ⁺ : 381.5.

^X H NMR (400 MHz, DMSO-s1 ₆ ) δ 9.07 (s, 1H), 8.66 (s, 2H), 7.84 (s, 1H), 6.87 (s, W), 6 .53 (d, J=8 Hz, 1H), 4.60-4.53 (m, 1H), 3.68 (s, ZI), 3.15 (dd, J=14, 5 Hz, 1H) , 3.01-2.95 (m, 1H), 2.80-2.76 (m, 4H), 2.61-2.57 (m, 4H), 1.98-1.90 (m, 4H).

2AF

Methyl yl)propanoate.

(2R)-2-{ [(1,2,3,5,6,7-hexahydro-8-indacen-4-yl)carbamoyl]amino}-3-(pyrazin-2-

IV: methyl (2B)-2-amino-3-(pyrazin-2-yl)propanoate hydrochloride.

General method A.

The crude product was purified by acid preparative HPLC to obtain the desired product as a white solid.

Yield = 3%.

MS ES ⁺ : 381.5.

Ή NMR (400 MHz, DMSO-th ₆ ) δ 8.60-8.55 (m, 2H), 8.52 (d, J=3 Hz, 1H), 7.88 (s, 1H), 6, 87 (s, 1H), 6.51 (d, J=8 Hz, 1H), 4.73-4.65 (m, 1H), 3.64 (s, ZH), 3.30-3.16 (m, 2H), 2.80-2.76 (m, 4H), 2.61-2.57 (m, 4H), 1.961.90 (m, 4H).

2AG

Ethyl 2-{[(1,2,3,5,6,7-hexahydro-8-indacen-4-yl)carbamoyl]amino}-3-(pyridazin-4-yl)propanoate.

IV: ethyl 2-amino-3-(pyridazin-4-yl)propanoate hydrochloride.

General method A.

The crude product was purified by acid preparative HPLC to obtain the desired product as a white solid.

Yield = 21%.

MS ES ⁺ : 395.4.

*H NMR (400 MHz, DMSO-th ₆ ) δ 9.14 (d, J=5 Hz, 1H), 9.11 (s, 1H), 7.83 (s, 1H), 7.56-7 .54 (m, 1H), 6.88 (s, 1H), 6.51 (d, J=8 Hz, 1H), 4.61-4.53 (m, 1H), 4.12 (q, J=1 Hz, 2H), 3.15 (dd, J=14.5 Hz, 1H), 3.06-3.0 (m, 1H), 2.80-2.76 (m, 4H), 2.62-2.58 (m, 4H), 1.98-1.90 (m, 4H), 1.19 (t, J=7 Hz, ZN).

2AN

Ethyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-8-indacen-4-yl)carbamoyl]amino}-3-(pyrimidin-2yl)propanoate.

IV: ethyl 2-amino-3-(pyrimidin-2-yl)propanoate dihydrochloride.

The racemic compound was synthesized according to the method detailed in Example 2VV. The racemate was separated by chiral HPLC to obtain the desired product as a white solid.

Yield = 14%.

MS ES ⁺ : 395.

-54044992 ¹ H NMR (400 MHz, DMSO-th ₆ ) δ 8.74 (d, J=5 Hz, 2H), 7.92 (s, 1H), 7.40 (t, J=5 Hz, 1H ), 6.87 (s, 1H),

6.49 (d, J=9 Hz, 1H), 4.85-4.69 (m, 1H), 4.05 (q, J=7 Hz, 2H), 3.32-3.29 (m , 2H), 2.78 (t, J=7 Hz, 4H), 2.71-2.56 (m, 4H), 2.04-1.83 (m, 4H), 1.10 (t, J=7 Hz, 3H).

2AI

Ethyl 2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(pyrimidin-4-yl)propanoate.

IV: ethyl 2-amino-3-(pyrimidin-4-yl)propanoate hydrochloride.

General method A.

The crude product was purified by acid preparative HPLC to obtain the desired product as a white solid.

Yield =10%.

MS ES+: 395.4.

1H NMR (400 MHz, DMSO-a ₆ ) δ 9.09 (d, J=1 Hz, 1H), 8.72 (d, J=5 Hz, 1H), 7.92 (s, 1H), 7 .49-7.42 (m, 1H), 6.87 (s, 1H), 6.53 (d, J=8 Hz, 1H), 4.73-4.66 (m, 1H), 4. 09 (q, J=7 Hz, 2H), 3.23-3.16 (m, 1H), 3.06-3.0 (m, 1H), 2.81-2.76 (m, 4H) , 2.64-2.58 (m, 4H), 1.98-1.88 (m, 4H), 1.15 (t, J=7 Hz, 3H).

2AJ

2-{[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)carbamoyl]amino}-3-(pyrimidin-2-yl)propanoic acid.

IV: 2-amino-3-(pyrimidin-2-yl)propanoic acid dihydrochloride.

General method A.

Yield = 26%.

MS ES+: 367.1.

¹ H NMR (300 MHz, DMSO-th ₆ ) δ 12.58 (br s, 1H), 8.73 (d, J=5 Hz, 2H), 7.89 (s, 1H), 7.38 (t, J=5 Hz, 1H), 6.86 (s, 1H), 6.39 (d, J=9 Hz, 1H), 4.77-4.68 (m, 1H), 2.80 -2.75 (m, 4H), 2.64-2.58 (m, 4H), 1.98-1.88 (m, 4H). 2 protons are shielded by a peak of water or DMSO.

Summary table of described structures

Example No. Structure Name 2A / NN Af sn ₃ \ / NN ¹¹ k—' o methyl 2-{[(1,2,3,5,6,7-hexahydrozind acen-4-yl)carbamoyl] amino } -3 -(3 hydroxyphenyl)propanoate

- 55 044992

2B & A HA Ζ Ν Ν DG CH ₃ \ NN ¹¹ <—' 0 Methyl 2-{ [(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl] amino } -3 -(2hydroxyphenyl)propanoate 2C /^° ZNN Tf sn ₃ \ / NN ¹¹ >—' o methyl 3-(3-acetylphenyl)-2-{[(1,2,3,5,6,7hexahydro-8-indacene-4- yl)carbamoyl] amino } propanoate 2D & A YES ZN ^N IT ^sn z \ / NN ¹¹ \--- ¹ O methyl (2K)-3-(4-cyanophenyl)-2{[(1,2,3,5,6,7-hexahydro-8-ind acene-4yl)carbamoyl] amino } propanoate 2E N \ / n n ¹¹ >—' o methyl (2K)-3-(3-cyanophenyl)-2{[(1,2,3,5,6,7-hexahydro-8-ind acen-4yl)carbamoyl] amino } propanoate 2F 1—k nA / \ ΐ \ Ml and A AA A A® Ζ Ν N Af CH ₃ \ / NN ¹¹ s—' 0 methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl] amino}-3 (pyridin-2-yl)propanoate 2G x Ax X1A Ax Z Ν N CH _E \--1 ⁿ no Methyl 2-{[(1,2,3,5,6,7-hexahydro-8indacen-4-yl)carbamoyl]amino}-3-[3(hydroxymethyl)phenyl]propanoate 2H l A ¹ » Ml S m^ AmA A A® Z 7^ Ν N CH ₃ \ NN ¹¹ k.—' o methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl ] amino } -3 (pyridin-3-yl)propanoate

- 56 044992

21 , ^Ν =\ Χ/ ^ν ~όη ₃ j5aX? / Ν Ν Tf sn ₃ \ / Η Η ¹¹ '—' ο methyl 2-{[(1,2,3,5,6,7-hexahydro-8- ind acen-4-yl)carbamoyl ] amino } -3 -[3 -(1 methyl-1 H-imidazol-5yl)phenyl]propanoate 2J Γ— ΝΗ j5a XT Ζ Ν Ν Tf CH ₃ \ / Η Η ¹¹ '--' 0 Methyl 2-{[(1,2,3,5,6,7-hekcahydro-sindacen-4-yl)carbamoyl]amino}-3-[3(1H-pyrazol-5-yl)phenyl]propanoate 2K )= ^ο I ζ^ ο =/ /=\ ? ο ^Ν ---- ^f s? methyl (2R)-2-{[(1,2,3,5,6,7-hexahydrozind acen-4-yl)carbamoyl] amino } -3 -(3 hydroxyphenyl)propanoate 2L M? J ΖχζΧ _Ί ·-^Ν ^ν 'Χζ ^Οχ ^ \ / Η Η ¹¹ '—' ο methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-sindacen-4-yl)carbamoyl]amino}-3-[3(1H-pyrazol-3-yl)phenyl ] propanoate 2M but X Λ V Ζ Ν ^Ν ΤΙ ^CH 3 \ / Η Η ¹¹ V-—' ο methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-sindacen-4-yl)carbamoyl]amino}-3-[3(hydroxymethyl)phenyl]propanoate 2Ν // ^ΝΗ 2 λ! α ΧΧ Ζ ν ν >Γ сн ₃ \ / Η Η ¹¹ '— ^J 0 methyl (2R)-3-amino-2-{[(1,2,3,5,6,7hexahydro-8-indacen-4yl)carbamoyl] amino } propanoate

- 57 044992

20 th, i—\ s ^v ch ₃ \ / n n H methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl ] amino } -4(methylsulfanyl) butanoate 2P O HgC^^NH Chl.X \ / H H II '—' o methyl (2R)-3-(3-aceτamidoφenyl)-2{[(1,2,3,5,6,7-hexahydro-8-ind acen-4yl)carbamoyl] amino } propanoate 2Q CH ₃ , 1 r—N XI a XI / T Ν N D CH ₃ \ / HH ¹¹ '---' 0 methyl 2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl] amino } -3 -(1 methyl-1H-pyrazol-4-yl)propanoate 2R χ l X \ / Η Η ¹¹ '----' 0 methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-sindacen-4-yl)carbamoyl]amino}-3-[3-(2- oxopyrrolidin-1-yl)phenyl] propanoate 2S sn ₃ °yi ix l iL < T ^NN p ^sn z \ / HH ¹¹ %—- ¹ 0 1,5-dimethyl (2R)-2-{[(l,2,3,5,6,7 hexahydro-s-indacen-4yl)carbamoyl] amino } pentanedioate 2T Oil I.o. ,ch _; 'ν ν γ< HH ¹¹ o ethyl 2-[({1H,2H,3H,6H,7H,8H,9Hcyclopenta[a]naphthalene-5yl} carbamoyl)amino] acetate

- 58 044992

2U n-nh o Cl I LT / N ν'>r CH ₃ \ / HH ¹¹ '—' o methyl (2R)-2-{[(1,2,3,5,6,7-hexahydrozind acen-4-yl)carbamoyl] amino } -3 - { 3 [(1H-pyrazol-3- yl)amino] phenyl } propanoate 2V n-nh A? HN ch A IL S LL,A _g? \° ⁿ \ / HH ¹¹ '—' o (2R)-2-{ [(1,2,3,5,6,7-hexahydrosindacen-4-yl)carbamoyl]amino}-3-[3(1H-pyrazol-3-yl)phenyl]propane acid 2W i—\ ⁰ dLL JL ,ch ₃ DA l Al Z ST N Ν'>r CH ₃ \ / HH AND 1,4-dimethyl (2R)-2-{[(l,2,3,5,6,7 hexahydro-s-indacen-4yl)carbamoyl]amino}butanedioate 2X IL I AA Ak .ch, ZNN AC\/HH ¹¹ 'o ethyl 2-{[(1,2,3,5,6,7-hexahydrosindacen-4-yl)carbamoyl] amino } acetate 2Y fL 2 Z Sr NN ^s CH, \ / HH AND >----' 0 methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl ] amino } -3 phenylpropanoate 2Z /Ch/ ^he Ol i lT ZN ν' CH ₃ \ / HH ¹¹ '— ^J o methyl (2R)-2-{[(l,2,3,5,6,7-geκcahydro-sind acen-4-yl)carbamoyl ] amino } -3 -(4hydroxyphenyl)propanoate 2AA IL 1 AA A\ Zk zA -0. -CH, ZNN >< Y \ HH ^{11 1} v—' o CH ₃ propan-2-yl 2-{[(1,2,3,5,6,7-hexahydros-ind acen-4-yl)carbamoyl]amino} acetate

- 59 044992

2BB ί—ί νη ₂ XX l ώ. /γ Ν Ν' γ CH ₃ \ / Η Η ¹¹ '— ^J 0 methyl (2H)-3-carbamoyl-2-{[(1,2,3,5,6,7hexahydro-8-indacene-4- yl)carbamoyl] amino } propanoate 2CC ry JO fX И ( ⁷ Z 7^ NN CH ₃ \ / NN ¹¹ ---' 0 methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl] amino } -3 (thiophen-2-yl)propanoate 2DD --- X fY S ί ZE ZE A. ZNN sn ₃ \ / NN ¹¹ methyl 2-{[(1,2,3,5,6,7-hexahydro-8- ind acen-4-yl)carbamoyl ] amino } -3 -(1Nimide azol-1 -yl)propanoate 2EE & A^o, Ζ ν ν сн _e \/НН ¹¹ ' ^—J о methyl 2-{[(1,2,3,5,6,7-hexahydro-8- indacen-4-yl)carbamoyl] amino } acetate 2FF )5a XX Z 7^ Ν N TE ^sn z \ / NN ¹¹ >—' o (2 S)-2-{ [(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl] amino } -3 phenylpropanoic acid 2GG J^aX? Z Ν N Tg ^sn z \ / NN ¹¹ '—' o methyl 2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl ] amino } -3 phenylpropanoate 2HH ex] ° ρ ^Χ '^ΐί:> ΉΗ ₂ XaJX Ж Z 7^ Ν Ν TE ^с з \ / Η Η ¹¹ -----' 0 methyl (2H)-3-(3-aminophenyl)-2{[(1,2,3,5,6,7-hexahydro-8-ind acen-4yl)carbamoyl] amino } propanoate 211 Ι--\ ΟΗ ₃ Хд а XX ζ Ν ν' Ττ ^сн \ / Η Η ¹¹ '— ¹ ο methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl ] amino } -3 methoxypropanoate

- 60 044992

2L A. / /He YES A Χίο. / Ν N SN _E \ / Η N ¹¹ '—' o methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl ] amino } -3 hydroxypropanoate 2KK ^ΗΝ ~l DD ΓΥ and \ / NN ¹¹ '--' 0 ethyl (2R)-2-{ [(1,2,3,5,6,7-hexahydro-8indacen-4-yl)carbamoyl]amino}-3-[3(1H-pyrazol-4-yl)phenyl ]propanoate 2LL gu A £1 ϊ l Av. X /He / ^NN TG \ / NN ¹¹ '' 0 (2R)-2-{[(l,2,3,5,6,7-geκcahydro-sind acen-4-yl)carbamoyl ] amino } -3 (pyridin-3-yl)propanoic acid 2MM g?· A L 1 u L o _Λ сн /у\ ^А / \ / НН ¹¹ '----' 0 2-methoxyethyl (2R)-2-{[(1,2,3,5,6,7hexahydro-8-indacene-4- yl)carbamoyl]amino}-3-(pyridin-3yl)propanoate 2NN D) /C fl S C \-J ⁿ n Ji i cyclobutyl (2R)-2-{[(l,2,3,5,6,7hexahydro-8-indacene-4- yl)carbamoyl]amino}-3-(pyridin-3yl)propanoate 200 V·Al\/NN ¹¹ V— ^/ 0 cyclopropylmethyl (2R)-2-{[(1,2,3,5,6,7hexahydro-8-indacene-4- yl)carbamoyl]amino}-3-(pyridine-3- yl)propanoate 2PP D) xS DA l DSch Z ^{Ν Ν} ιΓ Ar \ \ / NN ¹¹ \ / ---' O <7 cyclopentyl (2R)-2-{[(l,2,3,5,6,7hexahydro-8-indacene-4- yl)carbamoyl]amino}-3-(pyridin-3yl)propanoate 2QQ D l £ K Ν N CH ₃ \ / NN ¹¹ -----' 0 methyl (2R)-3-cyano-2-{[(1,2,3,5,6,7hexahydro-8-indacen-4yl)carbamoyl] amino } propanoate

- 61 044992

2RR XXI Z 7^ Ν Ν Y \ / HH ¹¹ G' 0 ethyl (2R)-2-{ [(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl] amino } -3 (pyridin-3-yl)propanoate 2SS ΓΛ ill £ Ay XX AX ^X a ^h Z 7^ ν ν' VJ ^HH S (2R)-2-{[(l,2,3,5,6,7-reKcarHapo-sind acen-4-yl)carbamoyl ] amino } -3 phenylpropanoic acid 2TT CH ₃ Ci] ⁰ AA XL X\ ^.o. ZNN CH ₃ VJ Η Η Π 3 methyl 2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl ] amino } -3 -(4methyl-1 H-pyrazol-1 -yl)propanoate 2UU A? A HA Ak \X-o ch ₃ 7^ Ν N If ³ / Η H ¹¹ hcA ^{0 3} CH ₃ ethyl (2R)-2-({[2.6-6Hc(nponaH-2- yl)phenyl]carbamoyl}amino)-3-(pyridin3-yl)propanoate 2VV l—Λ nX Hu o ^Et 3 ^NHCI DAL-X ⁰ ·-^ \ / HH ¹¹ '— ¹ o ethyl 2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl] amino } -3 (pyrimidin-2-yl)propanoate 2WW H-.C. A .X X< sn ₃ \ / HH ¹¹ methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-sindacen-4-yl)carbamoyl]amino}-3-(5methoxypyridin-3-yl)propanoate 2XX CH ₃ 1—λ NX / \ II N jli 11 ⁰ ХА X A___.ch ₃ Z 7^ Ν N AT \ / HH ¹¹ '—' o ethyl 2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl ] amino } -3 -(3 methyl-1,2,4-oxadiazol-5-yl) propanoate

- 62 044992

2YY /5 l.c? /γΑ N ⁴ ' CH ₃ \ / HH II '----' 0 methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl] amino} -3 (pyridazin-3-yl)propanoate 2ZZ ,____ ^-0 fl u As. A. JL A. .o. .ch. / y n A yk \ η h and X— ¹ 0 ethyl 2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl] amino }-3-(1,2-oxazol-4-yl)propanoate 2 AB 1—k n-° / \ Ji/ rY ϊ As JA JL A. .0. .ch. / 7^ NN 7< \ / η h AND X— ^J 0 ethyl 2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl] amino }-3-(1,2-oxazol-3-yl)propanoate 2AC AA A\ A/°4/ ^CH inn AA ^J \ / η h And X— ¹ o ethyl 2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl] amino } - 3 - (1,3 oxazol-2-yl)propanoate 2 AD LA Gn A As. Aks^N P 2 to As A _V A A·. A ^H ; N Ν'A< η h and A 0 НзС СНз ethyl (2R)-2-({ [6-(propan-2-yl)-2,3d ihydro-1 H-inden-5- yl]carbamoyl}amino)-3-(pyridin-3yl)propanoate 2AE ja JQ 1Ϊ S [^ A A. A vA ^0^ ANN A< СНз \ / η h and X— ¹ 0 methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl ] amino } -3 (pyrimidin-5-yl)propanoate 2AF A l ,^ ³ /уАг A γ cosh \ / η h and \— ¹ 0 methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl ] amino } -3 (pyrazin-2-yl)propanoate 2AG A? Xj XX a aX^ Z N N %< \ / η h AND X—' 0 ethyl 2-{[(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl] amino}-3 (pyridazin-4-yl)propanoate

- 63 044992

AN

2AI

2AJ

ethyl (2R)-2-{ [(1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl] amino} -3 (pyrimidin-2-yl)propanoate ethyl 2-{[ (1,2,3,5,6,7-hexahydro-8ind acen-4-yl)carbamoyl ] amino } -3 (pyrimidin-4-yl)propanoate

2-{[(1,2,3,5, 6,7-hexahydro-8-ind acen-4yl)carbamoyl]amino}-3-(pyrimidin-2yl)propanoic acid

Activity.

Determination of inhibitory activity in vitro.

The biological activity of the compounds of the present invention was determined using the assay described below.

PBMC analysis for IC50 determination.

The compounds of the present invention were tested for their inhibitory activity on IL-1 release upon activation of NLRP3 in peripheral blood mononuclear cells (PBMC).

PBMCs were isolated from buffy coats by density gradient centrifugation on Histopaque-1077 (Sigma, cat. no. 10771). The isolated cells were seeded into the wells of a 96-well plate and incubated for 3 hours with lipopolysaccharide (LPS). After replacing the medium, compounds of the present invention were added (one compound per well) and the cells were incubated for 30 minutes. Cells were then stimulated with either ATP (5 mM) or nigericin (10 μM) for 1 h, and cell culture medium from the wells was collected for further analysis.

The release of IL-1e into the medium was determined by quantifying IL-1e in the medium using the IL-1e enzyme-linked immunosorbent assay (ELISA) Ready-SET-Go!, eBioscience, cat. No. 88-7261-88. Briefly, in the first step, high affinity binding plates (Corning, Costar 9018 or NUNC Maxisorp Cat No. 44-2404) were coated overnight at 4°C with the specific capture antibody included in the kit (anti-human IL-1e ref. 14 -7018-68). The plates were then blocked with blocking buffer for 1 hour at room temperature (RT) and, after washing with buffer (PBS with 0.05% Tween 20), incubated with standard protein and culture medium. After 2 hours of incubation at room temperature, the plates were washed and incubated with the biotinylated detection antibody included in the kit (anti-human IL-1e Biotin ref. 337110-68) for 1 hour at room temperature. The plates were washed and incubated with HRPstreptavidin for 30 minutes at room temperature and washed again. The signal developed after the addition of 3,39,5,59-tetramethylbenzidine peroxidase (TMB) until color appeared and the reaction was stopped with 2M H ₂ SO ₄ . A microplate spectrophotometer (BioTek) was used to detect signals at 450 nm. The detection range of the IL-1e ELISA was 2–150 ng/mL.

Determination of IC ₅₀ values was previously performed using Graph Pad Prism software, and the measured IC ₅₀ values for the compounds of the present invention are shown in Table. 1 below.

- 64 044992

Table 1

Example No. PBMC, 1C ₅₀ , µM 2A 2.5 2B 0.06 2C 0.35 2D 1.4 2E 1.7 2F 4.2 2G 1.4 2H 0.88 21 0.96 2J 0.41 2K 1.3 2L 0.14 2M 0.38 2N 9.3 20 3.2 2P 0.14 2Q 2.4 2R 0.33 2S 3.5 2T 3.1 2U 0.31 2V 0.52 2W 2.9 2X 1.5

- 65 044992

2Y 1.4 2Z 0.10 2AA 1.8 2BB 32 2SS 0.5 2DD 6.2 2EE 3.7 2FF 9.1 2GG 6.4 2NN 4.4 211 5.3 2JJ 26 2 K K 0.33 2LL 2 2MM 2.9 2NN 0.6 200 0.47 2PP 0.24 2QQ <10 2RR 1.7 2SS 14 2TT 14 2UU 21 2VV 0.12 2WW 0.45 2XX 5.1 2YY 3.3 2ZZ 3.3 2AB 1.8 2AC 0.67 2AD 2.9 2AE 4.5 2AF 0.46 2AG 6.0 2 AN 0.036 2AI 6.8 2AJ 15

These results indicate that the compounds of the present invention are capable of inhibiting the release of IL-1e upon activation of the inflammasome.

Equivalents.

Details of one or more embodiments of the invention are set forth in the accompanying description. Although any methods and substances similar or equivalent to those described herein may also be used in the practice or study of the present disclosure,

- 66 044992 preferred methods and substances are described below. Other features, objects and advantages of the invention will be apparent from the description and claims. In the description and the accompanying claims, the singular forms also include the plural unless the context clearly indicates otherwise. Unless otherwise specified, all technical and scientific terms used herein have the same meaning as commonly understood by one skilled in the art to which this description relates. All patents and publications cited in this specification are incorporated by reference.

The above description is presented for purposes of illustration only and is not intended to limit the invention to the exact form described, but by the appended claims.

Claims (12)

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof: Where R ₁ is selected from an unsubstituted hexahydroindacene ring: ^# \ 7 where # denotes a bond with the nitrogen atom of formula (I); R2 represents H, R ₃ represents a C _1-4 alkyl-R ₇ , where R ₇ is selected from a 5 or 6 membered monocyclic aryl ring system containing no heteroatom or containing 1, 2 or 3 heteroatoms independently selected from oxygen, nitrogen and sulfur, and a ring the R ₇ system is unsubstituted or substituted with 1 substituent selected from C _1-6 alkyl, C _1-6 alkylhydroxy, OH, COCH ₃ , amino, cyano and R ₈ ; where R ₈ represents a 5 or 6 membered monocyclic heteroaryl ring containing 1 or 2 heteroatoms independently selected from oxygen, nitrogen and sulfur, where the specified ring is unsubstituted or substituted with C _1-6 alkyl or oxo; And R4 represents H, a C _1-6 alkyl or a C _3-6 monocyclic alkyl group. 2. A compound according to claim 1, wherein R ₃ is methyl-R ₇ or ethyl-R ₇ . 3. A compound according to claim 1 or 2, wherein R ₇ is a monocyclic aryl substituted with at least one hydroxyl group. 4. A compound according to claim 1 or 2, where R ₇ represents a monocyclic aryl with a cyano substituent. 5. A compound according to claim 1 or 2, where R ₇ represents a 5 or 6 membered monocyclic aryl ring containing 1, 2 or 3 heteroatoms independently selected from oxygen, nitrogen and sulfur. 6. A compound according to any of the preceding claims, wherein R4 represents C _1-4 alkyl or C _3-6 cycloalkyl. 7. A connection selected from the group consisting of: - 67 044992 Structure Name x5x XX ° ^H Z 7^ ^NN II sn ₃ \ / HH ¹¹ methyl 2—{[(1,2,3,5,6,7-hexahydro-sn dacen-4-yl)carbamoyl]amino}- 3-(3-hydroxyphenyl)propanoate XX a XX Z Ν N >T CH ₃ \ / HH ¹¹ ' o methyl 2-{ [(1,2,3,5,6,7-hexahydro-z-indacen-4-yl)carbamoyl]amino}-3 -(2-hydroxyphenyl)propanoate g>dCx XX a X°. XZ Eg NN E1 sn ₃ \ / HH ¹¹ '—' o methyl 3-(3-acetylphenyl)-2-{[(1,2,3,5,6,7-hexahydro8-indacen-4-yl)carbamoyl] amino}propanoate g\ Xa XX a XX ZNN ⁴ ER sn _e \ / HH ¹¹ \' o methyl (2R)-3 -(4-cyanophenyl)-2- {[(1,2,3,5,6, 7-hexahydro-3-indacen-4-yl)carbamoyl]amino}propanoate Ν\/n n ¹¹ '----' O methyl (2R)-3 -(3-cyanophenyl)-2-{[(1,2,3,5,6,7hexahydro-8-indacene-4- yl)carbamoyl] amino } propanoate 1--k M^X / \ ϊι 1 GT k Z 7^ ^NN IT ^sn z \ / Η Η ¹¹ ' O methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro -8-indacen-4yl)carbamoyl]amino}-3-(pyridin-2-yl)propanoate jd XXl°h XX aXJa ZNN ER CH ₃ \—/ ⁿⁿ 0 methyl 2-{ [(1,2,3, 5,6,7-hexahydro-8-indacen-4yl)carbamoyl]amino}-3-[3-(hydroxymethyl)phenyl]propanoate - 68 044992 \___j Η 0 Ν'' Η 0 0' ,Ν CH ₃ methyl (2K)-2-{[(1,2,3,5,6,7-hexahydro-8-indacen-4-yl) carbamoyl]amino}-3-(pyridin-3-yl)propanoate Ν = methyl 2—{[(1,2,3,5,6,7-hexahydro-8-indacene-4- „N CH ₃ yl)carbamoyl] amino }—3—[3—(1 -methyl- 1 H-imide azol-5yl)phenyl]propanoate \ ιί 0 JL II Ζ^/Ζ^ w γ° θΗ ₃ \___1 η Η 0 —Γ ΜΗ methyl 2—{[(1,2,3,5,6,7-hexahydro-8-indacene-4-λ,Ν yl )carbamoyl]amino}-3-[3-(1H-pyrazol-5yl)phenyl]propanoate Γ II 0 0. ,ΖγΖ 'CH _E \___/ Η Η ο he methyl (2K)-2-{[(1,2,3,5,6,7-hexahydro-8-indacen-4-yl)carbamoyl]amino}-3 -(3-hydroxyphenyl)propanoate Ν' 0. 'sn ₃ \____/ ^Η Η ο ί| methyl (2K)-2-{[(1,2,3,5,6,7-hexahydro-8-indacene-4- ΓΠ ο JI t y yl)carbamoyl]amino}-3-[3-(1H-pyrazol-3- /γΖ/ Zj Ζχ CH ^Ν ΝΗ yl)phenyl]propanoate \__/ Η Η ο no. methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-8-indacen-4-yl)carbamoyl]amino}-3-[3-(hydroxymethyl)phenyl]propanoate 0 oi ^сн ₃ ΖηΖ^Ν Ν' \___/ Η Η ο - 69 044992 0 methyl (2K)-3-(3-acetamidophenyl)-2-{[(1,2,3,5,6,7-n ₃ c' ^NH hexahydro-8-indacen-4yl)carbamoyl] amino } propanoate ° ( /ul' \___/ Η Η ^°'sn _e O CH ₃ methyl 2-{ [(1,2,3,5,6,7-hexahydro-8-indacene-4- o g Ν zCz ^N yl)carbamoyl] amino }-3-( 1 -methyl-1 H- pyrazol-4yl)propanoate γ__/ Η LG Η Y CH ₃ O methyl (2K)-2-{[(1,2,3,5,6,7-hexahydro-8-indacene-4- 0 N L yl)carbamoyl] amino }-3-[3-(2-oxopyrrolide in-1 yl)phenyl] propanoate Η Γ AU Ζ/^^Ν \__/ Η Ζ Η Y°Zh ₃ o N-NH methyl (2K)-2-{[(1,2,3,5,6,7-hexahydro-8-indacene-4- II A hn·^^ 1 Ί yl)carbamoyl]amino}-3-{3-[(1H-pyrazol-3yl)amino]phenyl} propanoate ⁰ 1 Ζ/Ζ^ζ rZ ] .0. f CH _E η ·· O N-NH (2R)-2-{ [(1,2,3,5,6,7-hexahydro-8-indacene-4- J // zZ yl)carbamoyl]amino}-3-[3-(1H-pyrazol-3yl)phenyl]propanoic acid Df(yZ >Z\ ^he H ¹¹ 0 >^ХХ ^0Н methyl (2R)-2-{[(l,2,3,5,6,7-heκcahydro-s-indacen-4- ⁰ X from yl)carbamoyl]amino}-3-( 4-hydroxyphenyl)propanoate /ug/ \___/ Η H if sn ₃ O - 70 044992 ΓΛ ΓΆ Ar s I ^x / 7^ ν ^Ν Γί сн ₃ \ / Η Η ¹¹ '--' 0 methyl (2K)-2-{[(1,2,3,5,6,7-hexahydro-8- indacen-4-yl)carbamoyl]amino}-3-(thiophen-2-yl)propanoate /--\ χΐ1Δ / 7^ Ν Ν ]< CH ₃ \ / Η Η ¹¹ '—' ο methyl 2—{[( 1,2,3,5,6,7-hexahydro-8-indacen-4yl)carbamoyl] amino} 3 (1 H-imidazol-1 yl)propanoate Γ|[ Ο /XJk Λ>Ο ₄ Ζ 7^ ν ν' γρ сн ₃ \ / Η Η ¹¹ '--' 0 methyl (2R)-3-(3-aminophenyl)-2-{[(1,2 ,3,5,6,7hexahydro-8-indacen-4-yl)carbamoyl] amino } propanoate ^ΗΝ- Λ ^ν ύζ ΓΥ 2 \ / Η Η ¹¹ '—' o ethyl (2R)-2-{ [(1, 2,3,5,6,7-hexahydro-8-indacen-4yl)carbamoyl]amino}-3-[3-(1H-pyrazol-4yl)phenyl]propanoate χ jQ ii 3 HDC X Α/°η Ζ ν ν yr \ Η Η ¹¹ '---- ¹ 0 (2R)-2-{ [(1,2,3,5,6,7-hexahydro-8-indacen-4-yl)carbamoyl]amino}-3 -(pyridin-3-yl)propanoic acid Χχ Gr ΑΆ Lk ,χΆ ^0' ^ ^sn Ζ Ν Ν τΓ Ο \ / Η Η ¹¹ '--' 0 2-methoxyethyl (2R)-2-{ [(1,2,3,5,6,7- hexahydro-8indacen-4-yl)carbamoyl]amino}-3-(pyridin-3yl)propanoate Χ> χθ £1 i /4 ^A N ^A N ^l V ^O VT VJ ^{Η n} ϊ U cyclobutyl (2R)-2-{ [(1,2,3,5,6,7-hexahydro-8indacen-4-yl )carbamoyl]amino}-3-(pyridin-3yl)propanoate ΛΥ Zy ^/ ^N' ^X ^'N ^V '°'·''' ^χΖ — V—/ ^{Η Η} ο cyclopropylmethyl (2R)-2-{[(1,2,3,5,6,7-hexahydro- 8indacen-4-yl)carbamoyl]amino}-3-(pyridin-3yl)propanoate - 71 044992 jQ γΎ S \ / η h ¹¹ 1 / '— ¹ o ' / cyclopentyl (2R)-2-{ [(1,2,3,5,6,7-hexahydro-sindacen-4-yl)carbamoyl ]amino}-3-(pyridin-3yl)propanoate jo jO XI a / γ N n'>< \ / HH ¹¹ <—' o ethyl (2R)-2-{ [(1,2,3,5,6 ,7-hexahydro-s-indacen-4yl)carbamoyl]amino}-3-(pyridin-3-yl)πpoπanoat CH ₃ o < ^N A As As ^,0. < ^NN V CH ₃ VJ HH P 3 methyl 2—{[(1,2,3,5,6,7-hexahydro-8-indacen-4yl)carbamoyl] amino }-3-(4-methyl-1 H- pyrazol-1 yl)propanoate A J- AJ\ A. ch, NN If ³ / Η H ¹¹ hcA ° ³ CH ₃ ethyl (2R)-2-({[2,6-6nc(nponaH-2-yl)phenyl]carbamoyl}amino)- 3-(pyridin-3yl)propanoate 1—λ ν'Ά·. Ay o ^e ' ₃ ^nhci AO-n-^nA ⁰ ^^, \ / Η Η ¹¹ '' 0 ethyl 2-{[(1,2,3,5,6,7-hexahydro-3-indacene-4- yl)carbamoyl]amino}-3-(pyrimidin-2-yl)propanoate H ₃ Cs Al O N ⁴ ' DS '' CHg \ / Η Η ¹¹ '—' o methyl (2R)-2-{[(1,2,3,5,6,7-hexahydro-8-indacen-4yl )carbamoyl]amino}-3-(5-methoxypyridin-3yl)propanoate CH ₃ GL 1 a 11 1 1 ⁰ AX X O\ /O. ,сн Ν Ν OT \ / НН ¹¹ '—' o ethyl 2-{[(1,2,3,5,6,7-hexahydro-8-indacen-4yl)carbamoyl]amino}-3-(3-methyl -1,2,4-oxac11a7o1-5yl)propanoate - 72 044992 Ua Z? ff'N N ⁴ ' UU ''CH ₃ \ / η η H '----' 0 methyl (2K)-2-{[(1,2,3,5,6,7-hexahydro-8-indacene -4yl)carbamoyl]amino}-3-(pyridazin-3-yl)propanoate,_____ ο LI i ζΜ JU JU J^ yu. ,cu. / 7^ Ν N UU \ / and and \--' 0 ethyl 2—{[(1,2,3,5,6,7-hexahydro-8-indacen-4-yl)carbamoyl]amino}-3 -(1,2-oxazol-4-yl)propanoate Г\ Ν- ⁰ . / \ rY ϊ K JU JU A, th, / 7^ Ν N UU \ / η h I '--- ^} 0 ethyl 2—{[(1,2,3,5,6,7-hexahydro-8- indacen-4-yl)carbamoyl]amino}-3-(1,2-oxazol-3-yl)propanoate >° I z Όί < P J ethyl 2—{[(1,2,3,5,6,7 -hexahydro-8-indacen-4-yl)carbamoyl]amino}-3-(1,3-oxazol-2-yl)propanoate Г~\\il X J^ ^Us^-N Ϊ1 2 U JU A -cl .sn> Y^N N'UU H H I /U 0 NZS SNz ethyl (2R)-2-({ [6-(propan-2-yl)-2,3-dihydro-1Ninden-5-yl]carbamoyl}amino)-3-(pyridin-3yl)propanoate JLL Λί fY S m^ X JU .JU vU A Ν N UU CH3 \ / η h I '---- ¹ 0 methyl (2R)-2-{[(1,2,3,5,6,7- hexahydro-8-indacen-4-yl)carbamoyl]amino}-3-(pyrimidin-5-yl)propanoate j5 a and rU Μ Y ^сНз \ / η h and \—ί o methyl (2R)-2-{ [(1,2,3,5,6,7-hexahydro-8-indacen-4yl)carbamoyl]amino}-3-(pyrazin-2-yl)propanoate I z °=\ z=\ O G z < V/o J ethyl 2-{ [(1,2,3,5,6,7-hexahydro-8-indacene-4- yl)carbamoyl]amino}-3-(pyridazin-4-yl)propanoate - 73 044992 N About SNz Ν About SNz Ν OH ethyl (2R)-2-{ [(1,2,3,5,6,7-hexahydro-s-indacen-4yl)carbamoyl]amino}-3-(pyrimidin-2-yl)πpoπanoate ethyl 2-{ [(1,2,3,5,6,7-hexahydro-8-indacen-4yl)carbamoyl]amino}-3-(pyrimidin-4-yl)propanoate 2-{[(1,2,3,5,6,7-hexahydro-8-ind acen-4yl)carbamoyl]amino}-3-(pyrimidin-2-yl)propanoic acid 8. A pharmaceutical composition containing a compound according to any of the preceding paragraphs or a pharmaceutically acceptable salt thereof in admixture with a pharmaceutically acceptable diluent or carrier. 9. A method of inhibiting the activity of the NLRP3 inflammasome in vitro or in vivo, wherein the method comprises contacting the cell with an effective amount of a compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof. 10. A method of treating a disease or disorder in which a patient in need of such treatment exhibits inflammasome activity, wherein said method comprises administering to said patient a therapeutically effective amount of a compound of any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof. I. The method of claim 10, wherein the disease or disorder is an autoinflammatory disorder, an autoimmune disorder, a neurodegenerative disease, or a malignancy. 12. The method of claim 11, wherein the malignancy is selected from gastrointestinal cancer, skin cancer, non-small cell lung cancer and colorectal adenocarcinoma. 13. The method of claim 11, wherein the disease or disorder is selected from cryopyrin-associated autoinflammatory syndrome (CAPS), including familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), chronic infantile neurological cutaneous articular syndrome (CINCA), multisystem neonatal inflammatory disease (NOMID), familial Mediterranean fever, non-alcoholic fatty liver disease (NAFLD), gout, rheumatoid arthritis, Crohn's disease, COPD, fibrosis, obesity, type 2 diabetes, multiple sclerosis, neuroinflammation arising from diseases associated with abnormal protein folding, Parkinson's disease, osteoarthritis, non-alcoholic steatohepatitis (NASH) and Alzheimer's disease.