1-(Sulfonyl)-N-phenylpyrrolidine-2-carboxamide derivatives and their uses

Abstract

1-(Sulfonyl)-N-phenylpyrrolidine-2-carboxamide derivatives and their uses The present invention relates to 1-(sulfonyl)-N-phenylpyrrolidine-2-carboxamide derivatives according to formula (I) for use in the prevention and / or treatment of a disease or disorder selected from autoimmune diseases, organ transplant rejection, cancer, and neurological diseases or disorders. The present invention further relates to new compounds encompassed by such formula (I).

Description

FIELD OF THE INVENTION

[0001] The present invention relates to medicinal chemistry. In particular, the present invention relates to 1-(sulfonyl)-N-phenylpyrrolidine aa-2-carboxamide derivatives of formula (I) for use as calcineurin inhibitors. The present invention further relates to new compounds falling within such formula (I). Background

[0002] The immune system is a complex network of cells and molecules that work together to protect the body from various diseases. Overactivity or inappropriate activity of the immune system is a serious and widespread medical problem. It contributes to acute and chronic immune disorders, such as allergic and atopic diseases, asthma, allergic rhinitis, allergic conjunctivitis, and atopic dermatitis, and in certain circumstances, the immune system also attacks and damages the body's own tissues, organs, and cells, resulting in autoimmune diseases (AD).

[0003] Nearly any part of the body can be targeted by the immune system, including the heart, brain, nerves, muscles, skin, eyes, lungs, gastrointestinal tract, and blood vessels. Given the variability in the body parts targeted by the immune system, there is a wide range of AD. Currently, over 100 different types of AD have been identified. Common ADs include rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis, type 1 diabetes, psoriasis, and celiac disease (see WO 2017 / 212018, pages 3-7, for a complete list of ADs). Indeed, AD affects 5-10% of the global population, with women being 2-10 times more likely to suffer from AD than men. Some forms of AD are among the top 10 leading causes of death in women aged 65 and over (Autoimmune Diseases - Modern Diseases, Study for the ENVI Commission, European Parliament, 2017, and references cited therein, available at http: / / www.europarl.europa.eu / cmsdata / 133620 / ENVI%202017-09%20WS%20Autoimmune%20diseases%20%20PE%20614.174%20(Publication).pdf).

[0004] Overactivity or inappropriate activity of the immune system also contributes to transplant rejection and graft-versus-host disease. Rejection occurs when the organ transplant recipient's immune system recognizes the donor's organ as foreign and attempts to eliminate it. Common transplant organs include, but are not limited to, kidneys, livers, hearts, lungs, and pancreases. Therefore, immunosuppressants prevent graft-versus-host reactions or tissue rejection during allografts or xenografts.

[0005] NFAT proteins (also called NFATc), a specific family of transcription factors, are expressed in immune cells and play a key role in eliciting immune responses. NFAT proteins are activated by calcineurin, which then induces the transcription of cytokine genes required for immune responses. The immunosuppressants cyclosporine A, tacrolimus (FK506), and voclosporin are potent inhibitors of cytokine gene transcription in activated immune cells and have been reported to inhibit calcineurin, thereby preventing it from activating NFAT. However, these drugs can exhibit nephrotoxicity and neurotoxicity after long-term use. Because calcineurin is ubiquitously expressed in many tissues, drug inhibition of calcineurin activity against substrates other than NFAT may contribute to the observed toxicity.

[0006] Calcineurin (CN or PPP3, formerly known as PP2B) is a Ca 2 CN is a serine / threonine protein phosphatase regulated by ATP and calmodulin. CN regulates development, memory, cardiac function, and immune responses by dephosphorylating various protein substrates, including the cytoplasmic nuclear factor of activated T cells (NFATc) family of transcription factors. NFATc proteins have been shown to be direct substrates of calcineurin. Dephosphorylated NFATc proteins are translocated into the nucleus, where they induce the expression of numerous cytokine and chemokine genes in cooperation with other transcription factors, resulting in T cell activation.

[0007] Calcineurin plays an important role in many physiological processes, including homeostasis, angiogenesis, adipogenesis, osteogenesis, chondrocyte differentiation, circulatory development, pancreatic beta cell proliferation, hair follicle cell differentiation and remodeling, and the activity of cells of the immune and nervous systems, including schizophrenia, diabetes, Down's syndrome, cardiac hypertrophy, and neurodegenerative diseases such as Alzheimer's disease, psychotic disorders, and epilepsy, among others (Erdmann, F et al., Calcineurin inhibitors: status quo and perspectives. BioMol Concepts, 2011, vol. 2, pp. 65-78; Kipanyula, MJ et al., The emerging roles of the calcineurin-nuclear factor of activated T-lymphocyte pathway in nervous system functions and diseases, J. Aging Research, 2016, Article ID 5081021, http: / / dx.doi.org / 10.1155 / 2016 / 5081021).

[0008] The immunosuppressants cyclosporine A (CsA) and tacrolimus (FK506) are the cornerstones of immunosuppressive therapy. These drugs sterically hinder the phosphatase activity of CN against NFATc and all other substrates by complexing with internal immunophilins and blocking the binding site of the peptide LxVP on CN. Both of these drugs have a narrow therapeutic window. Consequently, despite their benefits, serious side effects, including neurotoxicity, renal dysfunction, hypertension, and diabetes, are associated with long-term administration (review: Azzi, JR et al., Calcineurin inhibitors: 40 Years Later, Can't Live Without... J. Immunol. 2013 Dec 15, 191(12) 5785–5791; DOI: https: / / doi.org / 10.4049 / jimmunol.1390055).

[0009] Siebert, M et al. (Novel inhibitors of the Calcineurin / NFATc hub-aternatives to CsA and FK506? Cell Communication and Signaling, 2009, 7:25 doi:10.1189 / 1478-811X-7-25) and Erdmann, F. et al. (see above) review novel inhibitors of the calcineurin / NFATc signaling pathway. They further disclose that compounds that can distinguish between the inhibition of calcineurin and PPlases (peptidyl-prolyl cis-trans isomerases) as well as other substrates of NFATc and calcineurin may cause fewer side effects in clinical applications. To date, none of the substrates disclosed in these studies have been introduced into clinical trials.

[0010] According to prior art, the CN docking site of NFATc is mainly formed by the PxlxlT motif, the main anchor site, and the LxVP motif, which properly position the substrate phosphorylation residue relative to the catalytic active site (Li, H. et al., Interaction of Calcineurin with Substrates and Targeting Proteins. Trends in Cell Biology, 2011, 21(2), 91-103; Matsoukas, Minos et al. (2015). Identification of Small-Molecule Inhibitors of Calcineurin-NFATc signaling that mimic the PxIxIT motif of Calcineurin binding partners.Science Signaling.8.10.1126 / scisignal.2005918&Fig.S1 in supplementary information).

[0011] Perez-Riba et al. (WO2016207212 A1) and Matsoukas (supra) disclose non-peptide inhibitors of the calcineurin-NFAT signaling pathway that interrupt CNA-NFTAc interaction without blocking normal calcineurin phosphatase activity specifically through binding of the disclosed compounds to the PxlxlT binding domain of calcineurin.

[0012] WO 02 / 102380 A1 discloses mono- or bicyclic carbocycles and heterocycles useful as inhibitors of factor Xa for the treatment and prevention of thromboembolic disorders.

[0013] US2004 / 0006062 discloses silphonylaminovalerolactam and its derivatives useful as inhibitors of trypsin-like serine proteases, specifically factor Xa. These compounds do not contain the core structure contained in the compounds of formula (I) of the present invention.

[0014] EP3587416A1 discloses 2-oxopiperidin-3-yl derivatives and their use as calcineurin inhibitors. These compounds do not contain the core structure contained in the compounds of formula (I) of the present invention.

[0015] WO 02 / 00651 discloses inhibitors of trypsin-like serine protease enzymes, in particular factor Xa, as anticoagulants for the treatment and prevention of thromboembolic diseases.

[0016] US8455660B2 discloses the compound 1-(sulfonyl)-N-phenylpyrrolidine-2-carboxamide, which is potentially useful in drug discovery applications, and a method for its synthesis is also revealed.

[0017] US2012 / 0122950A1 discloses the compound 1-(sulfonyl)-N-phenylpyrrolidine-2-carboxamide, which has the potential to be used in drug discovery. The document discloses methods for obtaining the compounds and cites their potential use as anti-inflammatory or anticoagulant agents, among many other uses (cited in original). However, this anti-inflammatory activity is not supported by any examples and is only presented as a potential use. Therefore, this patent document is merely speculative, without supporting any potential use of the disclosed compounds.

[0018] Hanahan, D. and Weinberg, RA (2000). The Hallmarks of Cancer. Cell, 100(1), 57-70. DOI: 10.1016 / S0092-8674(00)81683-9. This article highlights the biological complexity of the disease and identifies six fundamental characteristics of cancer that go beyond inflammation. These characteristics include evasion of apoptosis, persistent replication, angiogenesis, invasion, metastasis, and resistance to growth inhibition.

[0019] Cooper, GS and Stroehla, BC (2003). The epidemiology of autoimmune diseases. Autoimmunity Reviews, 2(3), 119-125. DOI: 10.1016 / S1568-9972(03)00006-5. This review article provides insight into the diverse nature of autoimmune diseases, emphasizing distinct pathogenic mechanisms beyond inflammation, including genetic factors and immune dysregulation.

[0020] Therefore, there is a need to develop alternative strategies to interrupt the Ca2+-calcineurin-NFAT signaling pathway that are more specific for the calcineurin / NFAT pathway and overcome the problems associated with methods based on the use of currently available immunosuppressants. Furthermore, there is a need for immunosuppressants that selectively inhibit the calcineurin-NFAT interaction and do not inhibit the enzymatic activity of calcineurin toward other substrates.

[0021] The applicant of the present invention has surprisingly found that compounds of formula (I) compete for NFATc binding to CN through a site distinct from the above-mentioned PxlxlT- and LxVP-binding motifs in CN. The aforementioned pocket can be considered as a novel binding site for calcineurin inhibition. Summary of the invention

[0022] In a first aspect, the present invention relates to a 1-(sulfonyl)-N-phenylpyrrolidine-2-carboxamide derivative of formula (I) for use in the prevention and / or treatment of a disease or disorder, wherein the disease or disorder is selected from autoimmune diseases, organ transplant rejection, cancer, and neurological diseases or disorders.

[0023] In a second aspect, the present invention relates to new compounds encompassed by formula (I): [Brief explanation of the drawings]

[0024] FIG. 1 shows the effect of INK compounds (see references in the Examples) on NFATc activity in HEK293T cells.

[0025] FIG. 2 shows that INK compounds (see references in the Examples) do not affect CN phosphatase activity on pNPP (p-nitrophenyl phosphate) substrate.

[0026] FIG. 3 shows compounds that do not inhibit calcineurin phosphatase activity towards RII substrates.

[0027] FIG. 4 shows the synthetic scheme for compound SLB-605.

[0028] FIG. 5 shows the assay of compound SLB-605 against rhabdomyosarcoma cell lines (RD(a) and RH4(b)).

[0029] FIG. 6 shows the assay of compound SLB-605 against neuroblastoma cell lines (SKNBE2 (a) and SHSY5Y (b)).

[0030] FIG. 7 shows the assay of compound SLB-605 on osteoblast cell lines.

[0031] Figure 8 shows the assay of compound SLB-605 against medulloblastoma cell lines.

[0032] In a first aspect, the present invention relates to 1-(sulfonyl)-N-phenylpyrrolidine-2-carboxamide derivatives, also referred to by the term "compounds" of formula I,

[0033] [ka]

[0034] including stereoisomers and pharmaceutically acceptable salts thereof, R 1 is hydrogen; alkyl optionally substituted with Het; benzoyl optionally substituted with alkyl or cyano; or Het, R 2 is aryl optionally substituted with 1, 2, or 3 substituents each independently selected from alkyl, halo, cyano, amido, and carboxyl; benzyl optionally substituted with 1, 2, or 3 substituents each independently selected from alkyl, halo, cyano, amido, and carboxyl; Each Het as a group or part of a group is a monocyclic ring of 5 or 6 atoms; or a bicyclic ring structure consisting of a 6-membered ring fused to a 4-, 5-, or 6-membered ring; the rings (monocyclic or bicyclic) are each saturated, partially unsaturated, or fully unsaturated, and at least one of the rings contains from 1 to 4 heteroatoms each independently selected from nitrogen, oxygen, and sulfur, and any of the rings (monocyclic or bicyclic) is selected from halo, hydroxyl, nitro, cyano, C 1~6 Alkyl, and C 1~6and optionally substituted with 1, 2, or 3 substituents each independently selected from the group consisting of alkoxy.

[0035] In a preferred embodiment, in the compound of formula (I) used as a calcineurin inhibitor as shown above, R 1 is hydrogen; C optionally substituted with Het 1~6 Alkyl; C 1~6 benzoyl optionally substituted with alkyl or cyano; or Het, R 2 is C 1~6 aryl optionally substituted with 1, 2, or 3 substituents each independently selected from alkyl, halo, cyano, amido, and carboxyl; or C 1~6 benzyl optionally substituted with 1, 2, or 3 substituents each independently selected from alkyl, halo, cyano, and carboxyl; Each Het as a group or part of a group is a monocyclic ring with 5 or 6 atoms; or a bicyclic ring structure consisting of a 6-membered ring fused to a 5-membered or 6-membered ring, each of which is saturated, partially unsaturated, or fully unsaturated, and at least one of the rings contains 1 to 4 heteroatoms each independently selected from nitrogen, oxygen, and sulfur, and any of the rings is selected from halo, hydroxyl, cyano, C 1~6 and optionally substituted with 1, 2, or 3 substituents each independently selected from the group consisting of alkyl.

[0036] In a further preferred embodiment, in the compound of formula (I) used as a calcineurin inhibitor as shown above, R 1 is hydrogen; methyl optionally substituted with Het; or benzoyl optionally substituted with cyano, R 2is aryl optionally substituted with one, two, or three substituents each independently selected from methyl, halo, cyano, amido, and carboxyl; or benzyl optionally substituted with one, two, or three substituents each independently selected from methyl, halo, and cyano; Each Het as a group or part of a group is a bicyclic ring structure consisting of a 5-membered ring or a 6-membered ring fused to a 6-membered ring, each of which is partially unsaturated, and at least one of which contains 1 to 4 heteroatoms each independently selected from nitrogen, oxygen, and sulfur.

[0037] In a further preferred embodiment, in the compound of formula (I) used as a calcineurin inhibitor as shown above, R 1 teeth,

[0038] [ka]

[0039] [ka]

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[0042] and

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[0044] Selected from R 2 teeth,

[0045]

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[0055]

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[0057] [ka]

[0058] is selected from.

[0059] As used above and hereinafter, the following definitions apply unless otherwise stated.

[0060] The term "halo" is generic to fluoro, chloro, bromo, and iodo.

[0061] The terms "heterocyclic ring" or "heterocycle" are used interchangeably herein and refer to any compound in which multiple atoms form a ring via multiple covalent bonds, and the ring contains at least one atom other than a carbon atom. Heterocyclic bases are specifically contemplated to include 4-, 5-, and 6-membered rings containing at least 1 to 4 heteroatoms, each independently selected from nitrogen, oxygen, and sulfur, as non-carbon atoms (e.g., imidazole, pyrrole, triazole, dihydropyrimidine). Because heterocycles can be fused (i.e., covalently bonded) to other rings or heterocycles, the terms "fused heterocycle" or "fused heterocyclic base" are further contemplated to be used herein. Fused heterocycles include 5-membered rings fused to 6-membered rings (e.g., purine, pyrrolo[2,3-d]pyrimidine), fused to other 6- or more-membered rings (e.g., pyrido[4,5-d]pyrimidine, benzodiazepine). It is specifically contemplated that the heterocyclic base may comprise a six-membered ring fused to a heterocyclic base. It is further contemplated that the heterocyclic base may be aromatic and may contain one or more double or triple bonds. It is further contemplated that the heterocyclic base and the fused heterocyclic ring may be further substituted at one or more positions. Any one of the rings may be substituted with a halogen, hydroxy, nitro, cyano, carboxyl, C 1~6 Alkyl, C 1~6 Alkoxy, C 1~6Alkoxy C 1~6 Alkyl, C 1~6 Alkylcarbonyl, Amino, Mono C 1~6 Alkylamino or diC 1~6 Alkylamino, azide, mercapto, polyhalo C 1~6 Alkyl, Polyhalo C 1~6 Alkoxy, and C 3~7 Optionally substituted with 1, 2, or 3 substituents each independently selected from the group consisting of cycloalkyl.

[0062] "Het" is defined herein as a group or part of a group that is a monocyclic ring with 5, 6, or 7 ring atoms optionally fused to a bicyclic ring structure, including aryl, or a 5-, 6-, or 7-membered ring bonded to a 4-, 5-, or 6-membered ring; the rings are each saturated, partially unsaturated, or fully unsaturated; at least one of the rings contains from 1 to 5 heteroatoms each independently selected from nitrogen, oxygen, and sulfur; any of the rings may be selected from the group consisting of OH; F; Cl; Br; I; methyl; an optionally substituted alkyl chain; an optionally substituted alkenyl chain; an optionally substituted alkynyl chain; an optionally substituted cycloalkyl; an optionally substituted cycloalkenyl; an optionally substituted cycloalkynyl; OR 6 ;NR 6 R 7 ;CN;COR 6 ;CONR 6 R 7 ;CO2R 6 ;C(S)OR 6 ;OCONR 6 R 7 ;OCOR 6 ;OCO2R 6 ;OC(S)OR 6 ;N3;NO2;NHCONR 6 R 7 ;NHCOR 6 ;NR 6 CO2R 7 ;NHCO2R 6 ;NHC(S)OR 6 ;NO2;SR 6 ;SO3H;SO2R6 ;SO2NR 6 R 7 Optionally substituted with 1, 2, 4, or 4 substituents each independently selected from the group consisting of: an aryl; a Het; an optionally substituted alkyl, alkenyl, alkynyl chain, an aryl, or an optionally substituted aryl bonded to a Het; and an optionally substituted heterocycle bonded to an optionally substituted alkyl, alkenyl, or alkynyl chain.

[0063] The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that are interconvertible via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via migration of a proton, such as keto-enol and imine-enamine isomerizations. Valence tautomers include interconversions via reorganization of some of the bonding electrons.

[0064] For therapeutic use, salts of compounds of formula (I) are compounds whose counterions are pharmaceutically acceptable. However, salts of acids and chlorides that are not pharmaceutically acceptable may also find use, for example, in the preparation or purification of pharmaceutically acceptable compounds. All salts, whether pharmaceutically acceptable or not, are within the scope of the present invention.

[0065] The aforementioned pharmaceutically acceptable acid and base addition salts are intended to include therapeutically effective non-toxic acid and base addition salt forms that the compounds of Formula (I) can form. Pharmaceutically acceptable acid addition salts can be conveniently obtained by treating the base form with such a suitable acid. Examples of suitable acids include inorganic acids such as hydrohalic acids, e.g., hydrochloric acid or hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; and organic acids such as acetic acid, propanoic acid, hydroxyacetic acid, lactic acid, pyruvic acid, oxalic acid (i.e., ethanedioic acid), malonic acid, succinic acid (i.e., butanedioic acid), maleic acid, fumaric acid, malic acid (i.e., hydroxybutanedioic acid), tartaric acid, methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, cyclamic acid, salicylic acid, p-aminosalicylic acid, pamoic acid, and the like.

[0066] Conversely, such salt forms can also be converted into the free base form by treatment with an appropriate base.

[0067] Compounds of formula (I) containing an acidic proton can also be converted into non-toxic metal or amine addition salt forms by treatment with appropriate organic and inorganic bases. Suitable base salt forms include, for example, ammonium salts, alkali and alkaline earth metal salts such as lithium salts, sodium salts, potassium salts, magnesium salts, calcium salts, etc.; salts with organic bases such as benzathine, N-methyl-D-glucamine, hydrabamine, etc.; and salts with amino acids such as arginine and lysine.

[0068] The term "addition salts" as used above also comprises solvates which the compounds of formula (I) or any of the salts thereof are able to form. Such solvates are, for example, hydrates, alcoholates and the like.

[0069] The term "quaternary amine" as used herein defines a quaternary ammonium salt that can be formed by reacting a basic nitrogen of either a compound of Formula (I) or a compound of Formula (I) with a suitable quaternizing agent, such as an optionally substituted alkyl halide, aryl halide, or arylalkyl halide, e.g., methyl iodide or benzyl iodide. Other reactants with good leaving groups can be used, such as alkyl trifluoromethanesulfonate, alkyl methanesulfonate, and alkyl p-toluenesulfonate. Quaternary amines have a positively charged nitrogen. Pharmaceutically acceptable counterions include chloro, bromo, iodo, trifluoroacetate, and acetate. The counterion of choice can be introduced using ion exchange resins.

[0070] The N-oxide forms of the compounds are intended to include compounds of formula (I) wherein one or more nitrogen atoms have been oxidized to the so-called N-oxides.

[0071] It will be understood that the compounds of formula (I) may possess metal binding, chelating and complex-forming properties and may therefore exist as metal complexes or metal chelates, and such metallated derivatives of the compounds of formula (I) are intended to be included within the scope of the present invention.

[0072] Some of the compounds of formula (I) may also exist in tautomeric forms, and such forms, although not explicitly indicated in the above formula, are intended to be included within the scope of the present invention.

[0073] The compounds described herein may have asymmetric centers and may occur as racemates, racemic mixtures, individual diastereomers, or enantiomers, and all isomeric forms are intended to be included in the present invention. Compounds of the present invention having chiral centers may exist in and be separated into optically active racemic forms.

[0074] The term "alkyl" as used herein refers to any straight, branched, or cyclic hydrocarbon in which all carbon-carbon bonds are single bonds. An alkyl chain may be any of the following: OH; NH; F; Cl; Br; I; methyl; an optionally substituted alkyl chain; an optionally substituted alkenyl chain; an optionally substituted alkynyl chain; an optionally substituted cycloalkyl; an optionally substituted cycloalkenyl; an optionally substituted cycloalkynyl; OR 6 ;NR6R7;CN;COR6;CONR6R7;CO2R6;C(S)OR6;OCONR6R7;OCOR6;OCO2R6;OC(S)OR6;N3;NO2;NHCONR6R7;NHCOR6;NR6CO2R7;NHC2R6;NHC(S)OR6;NO2;SR6;SO3H;SO2R6;SO2NR6R7;aryl;Het; optionally substituted alkyl, alkenyl, alkynyl chain, aryl, or optionally substituted aryl bonded to Het; and optionally substituted alkyl, alkenyl, or optionally substituted heterocycle bonded to an alkyl chain.

[0075] The terms "alkenyl" and "optionally substituted alkenyl" are used interchangeably herein and refer to any straight-chain, branched, or cyclic alkyl with at least one carbon-carbon double bond. An alkenyl chain is selected from the group consisting of OH; NH; F; Cl; Br; I; methyl; optionally substituted alkyl chains; optionally substituted alkenyl chains; optionally substituted alkynyl chains; optionally substituted cycloalkyl; optionally substituted cycloalkenyl; optionally substituted cycloalkynyl; OR; NR; R; CN; COR; CONR; R; C(S)OR; OCONR; R; OCOR; OCO; OC(S)OR; N3; NO2; NHCONR6R7; NHCOR6; NR6CO2R7; NHC2R6; NHC(S)OR6; NO2; SR6; SO3H; SO2R6; SO2NR6R7; aryl; Het; optionally substituted alkyl, alkenyl, alkynyl chain, aryl, or optionally substituted aryl bonded to Het; optionally substituted heterocycle bonded to an optionally substituted alkyl, alkenyl, or alkynyl chain.

[0076] Furthermore, "alkynyl" and "optionally substituted alkynyl" are used interchangeably herein and refer to any straight, branched, or cyclic alkyl or alkenyl with at least one carbon-carbon triple bond. An alkynyl chain is selected from the group consisting of OH; NH; F; Cl; Br; I; methyl; optionally substituted alkyl chains; optionally substituted alkenyl chains; optionally substituted alkynyl chains; optionally substituted cycloalkyl; optionally substituted cycloalkenyl; optionally substituted cycloalkynyl; OR; NR; R; CN; COR; CONR; R; C(S)OR; OCONR; R; OCOR; OCO; OC(S)OR; N3; NO2; NHCONR6R7; NHCOR6; NR6CO2R7; NHC2R6; NHC(S)OR6; NO2; SR6; SO3H; SO2R6; SO2NR6R7; aryl; Het; optionally substituted alkyl, alkenyl, alkynyl chain, aryl, or optionally substituted aryl bonded to Het; optionally substituted heterocycle bonded to an optionally substituted alkyl, alkenyl, or alkynyl chain.

[0077] The term "aryl" as used herein refers to a group or part of a group of phenyl, naphthalenyl, anthracenyl, phenanthrenyl, biphenyl optionally fused to Het, and is, respectively, OH; F; Cl; Br; I; methyl; an optionally substituted alkyl chain, an optionally substituted alkenyl chain, an optionally substituted alkynyl chain, an optionally substituted cycloalkyl; an optionally substituted cycloalkenyl; an optionally substituted cycloalkynyl; OR 6 ;NR 6 R 7 ;CN;COR 6 ;CONR 6 R 7 ;CO2R 6 ;C(S)OR 6 ;OCONR 6 R 7 ;OCOR 6 ;OCO2R 6 ;OC(S)OR6 ;N3;NHCONR 6 R 7 ;NHCOR 6 ;NR 6 CO2R 7 ;NHCO2R 6 ;NHC(S)OR 6 ;NO2;SR 6 ;SO3H;SO2R 6 ;SO2NR 6 R 7 ; aryl; Het; optionally substituted aryl attached to an optionally substituted alkyl, alkenyl, alkynyl chain, aryl, or Het; optionally substituted with 1, 2, 3, or 4 optionally substituted substituents attached to an optionally substituted alkyl, alkenyl, or alkynyl chain.

[0078] As used herein, the term "substituted" refers to the replacement of an atom or chemical group (e.g., H, NH, or OH) with a functional group, and in particular functional groups include nucleophilic groups (e.g., -NH, -OH, -SH, -NC, etc.), electrophilic groups (e.g., C(O)OR, C(X)OH, etc.), polar groups (e.g., -OH), non-polar groups (e.g., aryl, alkyl, alkenyl, alkynyl, etc.), ionic groups (e.g., -NH, + ), and halogens (e.g., -F, -Cl), and all chemically reasonable combinations thereof. Accordingly, the terms "functional group" and "substituent" are used interchangeably herein and are intended to include nucleophilic groups (e.g., -NH, -OH, -SH, -NC, -CN, etc.), electrophilic groups (e.g., C(O)OR, C(X)OH, C(halogen)OR, etc.), polar groups (e.g., -OH), non-polar groups (e.g., aryl, alkyl, alkenyl, alkynyl, etc.), ionic groups (e.g., -NH + ), and halogens.

[0079] Functional groups such as -OH and -NH may incorporate protecting groups (abbreviated as PG), such as those known to those skilled in the art (GREENE, PROTECTIVE. GROUPS IN ORGANIC. SYNTHESIS, 4th ed., PETER GMWUTS. AND THEODORA W. GREENE, 2007, Wiley-Interscience). For example, hydroxyl protection, including 1,2-diols (Greene, supra, pp. 16-366), can take the form of ethers, esters, cyclic acetals, cyclic ketals, and silyl derivatives, such as methyl ethers, methoxymethyl ethers, methylthiomethyl ethers, t-butylthiomethyl ethers, (phenyldimethylsilyl) ethers, benzyloxymethyl ethers, p-methoxybenzyloxymethyl ethers, p-nitrobenzyloxymethyl ethers, o-nitrobenzyloxymethyl ethers, (4-methoxyphenoxy)methyl ethers, guaiacol methyl ethers, t-butoxymethyl ethers, 4-pentenyloxymethyl ethers, siloxymethyl ethers, 2-methoxyethoxymethyl ethers, 2,2,2-trimethyl ... chloroethoxymethyl ether, bis(2-chloroethoxy)methyl ether, 2-(triethylsilyl)ethoxymethyl ether, methoxymethyl ether, tetrahydropyranyl ether, 3-bromotetrahydropyranyl ether, tetrahydrothiopyranyl ether, 1-methoxycyclohexyl ether, 4-methoxytetrahydropyranyl ether, 4-methoxytetrahydrothiopyranyl ether, 4-methoxytetrahydrothiopyranyl S,S-dioxide ether, 1-[(2-chloro-4-methyl)phenyl]-4-methoxypiperidin-4-yl ether, 1,4-dioxan-2-yl ether, 1,4-dioxan-2-yl ether, tetrahydrothiofuranyl ether, 2,3,3a,4,5,6,7,7a-octahydro-7,8,8-trimethyl-4,7-Methanobenzofuran-2-yl ether, 1-ethoxyethyl ether, 1-(2-chloroethoxy)ethyl ether, 1-hydroxyethyl ether, 2-bromoethyl ether, 1-[2-(trimethylsilyl)ethoxy]ethyl ether, 1-(2-cyanoethoxy)ethyl ether, prenyl ether, cinnamyl ether, propargyl ether, p-nitrophenyl ether, 1-methyl-1-methoxyethyl ether, 1-methyl-1-benzyloxyethyl ether, 1-methyl-1-benzyloxy-2-fluoroethyl ether, 2,2,2-trichloroethyl ether, 2-trimethylsilylethyl ether, 2-(phenylselenyl)ethyl ether, t-butyl ether, allyl ether, p-chlorophenyl ether, p-methoxyphenyl ether, 2,4-dinitrophenyl ether, benzyl ether, p-methoxybenzyl ether, 3,4-dimethoxybenzyl ether, 2,6-dimethoxybenzyl, o-nitrobenzyl ether , p-nitrobenzyl ether, p-bromobenzyl ether, p-chlorobenzyl ether, 2,6-dichlorobenzyl ether, 2,4-dinitrobenzyl ether, fluorous benzyl ether, trimethylsilylxylyl ether, p-phenylbenzyl ether, cumyl ether, p-azidobenzyl ether, 2,6-difluorobenzyl ether, p-cyanobenzyl ether, p-phenylbenzyl ether, 2-picolyl ether, 4-picolyl ether, 3-methyl-2-picolyl N-oxide ether, diphenyl methyl ether, p,p'-dinitrobenzhydryl ether, 5-dibenzosuberyl ether, triphenyl methyl ether, α-naphthyl diphenyl methyl ether, p-methoxyphenyl diphenyl methyl ether, di(p-methoxyphenyl)phenyl methyl ether, tri(p-methoxyphenyl)phenyl methyl ether, 4-(4'-bromophenacryloxyphenyl)diphenyl methyl ether, 4,4',4"-tris(4,5-Dichlorophthalimidophenyl)methyl ether, pentadienylnitrobenzyl, p-azidobenzyl ether, p-(methylsulfinyl)benzyl ether, 2-naphthylmethyl ether, 2-quinolinylmethyl ether, 1-pyrenylmethyl ether, 4-methoxydiphenylmethyl ether, 4-phenyldiphenylmethyl ether, α-naphthyldiphenylmethyl ether, p-methoxyphenyldiphenylmethyl ether, anthryl ether, 9-phenylthioxanthyl ether, etc.; trimethylsilyl ether, triethylsilyl ether, triisopropylsilyl ether, dimethylhexylsilyl ether, 2-norbornyldimethylsilyl ether, t-butyldimethylsilyl ether, t-butyldiphenylsilyl ether, tribenzylsilyl ether, tri-p-xylylsilyl ether, triphenylsilyl ether, diphenylmethylsilyl ether, di-t-butylmethylsilyl ether, bis(t-butyl)-1-pyrene Silyl ethers such as 2-hydroxystyryl ether, tris(trimethylsilyl)silyl ether, (2-hydroxystyryl)dimethylsilyl ether, t-butoxydiphenylsilyl ether, 1,1,3,3-tetraisopropyl-3-[2-(triphenylmethoxy)ethoxy]disiloxan-1-yl ether, and fluorous silyl ethers; formates, benzoyl formates, acetates, chloroacetates, dichloroacetates, trichloroacetates, and trichloroacetimides. Date, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, phenoxyacetate, p-chlorophenylacetate, phenylacetate, diphenylacetate, 3-phenylpropionate, bisfluoropropanoyl ester, 4-pentenoate, levulinate, pivalate, adamantoate, crotonate, 4-methoxycrotonate, benzoate, p-phenylbenzoate, mesitoate, 4-bromobenzoate, 2,5-Difluorobenzoate, p-nitrobenzoate, picolinate, nicotinate, 2-(azidomethyl)benzoate, 4-azidobutyrate, (2-azidomethyl)phenylacetate, 2-{[(tritylthio)oxy]methyl}benzoate, 2-(allyloxy)phenylacetate, 2-(prenyloxymethyl)benzoate, 4-benzyloxybutyrate, 4-trialkylsiloxybutyrate, 4-acetoxy-2,2-dimethylbutyrate, 2,2-dimethyl-4-pentanoate, 2-iodobenzoate, 4-nitro-4-methylpentanoate, o-(dibromomethyl)benzoate, 2-formylbenzenesulfonate, 4-( Methylthiomethoxy)butyrate, 2-(methylthiomethoxymethyl)benzoate, 2-(chloroacetoxymethyl)benzoate, 2-[(2-chloroacetoxy)ethyl]benzoate, 2-[2-(benzyloxy)ethyl]benzoate, 2-[2-(4-methoxybenzyloxy)ethyl]benzoate, 2,6-dichloro-4-methylphenoxyacetate, 2,6-dichloro-4-(1,1,3,3-tetramethylbutyl)phenoxyacetate, chlorodiphenylacetate, isobutyrate, monosuccinate, tiglotte, o-(methoxycarbonyl)benzoate, p-benzoate, α-napthoate, nitrate, alkyl N,N,N',Esters such as N'-tetramethylphosphorodiamidate and 2-chlorobenzoate; sulfonates such as sulfates, allyl sulfonates, methanesulfonates, benzyl sulfonates, tosylates, and 2-trifluoromethyl sulfonates; alkyl methyl carbonates, methoxymethyl carbonates, 9-fluoromethyl carbonates, ethyl carbonates, bromoethyl carbonates, 2-(trimethylsilyl)ethyl carbonates, isobutyl carbonates, t-butyl carbonates, vinyl carbonates, allyl carbonates, and propargyl carbonates. Carbonates such as thiocarbonate, p-nitrophenyl carbonate, benzyl carbonate, 2-dansylethyl carbonate, phenacyl carbonate, methyldithiocarbonate, and S-benzylthiocarbonate; carbamates such as dimethylthiocarbamate and N-phenylcarbamate; methylene acetal, ethylidene acetal, t-butyl methylidene acetal, 1-t-butylethylidine ketal, 1-phenylethididene ketal, 2-(methoxycarbonyl)ethylidene acetal, 2-(t-butylcarbonyl)ethylidene acetal, phenyl Cyclic acetals and ketals such as phenylsulfonylethylidene acetal, 3-(benzyloxy)propylidene acetal, isopropylidene acetal or acetonide, cyclopentylidene acetal, benzylidene acetal, p-methoxybenzylidene acetal, mesitylene acetal, naphthaldehyde acetal, 9-anthracene acetal, and benzophenone ketal; chiral ketones such as camphor ketal and menthone ketal; methoxymethylene acetal, ethoxymethylene acetal, dimethoxymethylene orthoester, methylene Cyclic orthoesters such as methyldisiloxane orthoesters, phthalide orthoesters, 1,2-dimethoxyethylidene orthoesters, 2-oxacyclopentylidene orthoesters, butane 2,3-bisacetal, cyclohexane-1,2-diacetal, and dispiroketal; di-t-butylsilylene group, 1,3-(1,1,3,3-tetraisopropyldisiloxanylidene) derivatives, 1,1,3,3-tetra-t-butoxydisiloxanylidene derivatives, methylene-bis-(diisopropylsilanoxanylidene), 1,1,4,4-tetraphenyl-1,Silyl derivatives such as 4-disilanilidene, o-xylyl ether, and 3,3'-oxybis(dimethoxytrityl) ether; cyclic carbonates; and cyclic borates such as methyl boronate and ethyl boronate, but are not limited to these.

[0080] The term "optionally substituted" when referring to alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl, and heterocycle includes oxo, ethylenedioxy, alkanoyloxy, alkoxy, alkylthio, carboxyl, halogen, thienyl, acetyl, 1-oxopropyl, 2-oxopropyl, 2-oxobutyl, 3-oxobutyl, 3-oxopentyl, 4-oxopentyl, 4-oxohexyl, 5-oxohexyl, ethylenedioxymethyl, 1,1-ethylenedioxyethyl, 2,2-ethylenedioxyethyl, 1,1-ethylenedioxypropyl, 2,2-ethylenedioxypropyl, 3,3-ethylenedioxypropyl, 1,1-ethylenedioxybutyl, 2,2-ethylenedioxybutyl, 3,3-ethylenedioxybutyl, 4,4-ethylenedioxybutyl, 3,3-ethylenedioxypentyl, 4,4-ethylenedioxyhexyl, 5,5-ethylenedioxyhexyl, acetyloxymethyl, 2-acetyloxyethyl, 3-acetyloxypropyl, 3-acetyloxybutyl, 4-acetyloxybutyl, 3-propynyloxybutyl, 3-butyryloxybutyl, 3-valeryloxypentyl, 3-hexanoylhexyl, 4-acetyloxypentyl, 5-acetyloxypentyl, 4-acetyloxyhexyl, 5-acetyloxyhexyl, 6-acetyloxyhexyl, methoxymethyl, ethoxymethyl, propoxymethyl, butoxymethyl, pentyloxymethyl, hexyloxymethyl, 1-methoxyethyl, 2-methoxyethyl, 2-ethoxyethyl, 2-propoxyethyl, 2-butoxyethyl, 2-pentyloxyethyl, 2-hexyloxyethyl, 3-methoxypropyl, 3-ethoxypropyl, 2-methoxybutyl, 4-ethoxybutyl, 3-methoxypentyl, 5-ethoxypentyl, 4-methoxyhexyl The invention is intended to encompass the group including methyl, 6-ethoxyhexyl, methylthiomethyl, ethylthiomethyl, propylthiomethyl, butylthiomethyl, pentylthiomethyl, hexylthiomethyl, 1-methylthioethyl, 2-methylthioethyl, 2-ethylthioethyl, 2-methylthiopropyl, 3-methylthiopropyl, 3-ethylthiobutyl, 4-butylthiobutyl, 5-methylthiopentyl, 6-ethylthiohexyl, carboxymethyl, 1-carboxyethyl, 2-carboxyethyl, 2-carboxypropyl, 3-carboxypropyl, 4-carboxybutyl, 5-carboxypentyl, 6-carboxyhexyl, fluoromethyl, bromomethyl, chloromethyl, iodomethyl, 2-chloroethyl, 2-bromopropyl, 3-iodopropyl, 4-fluorobutyl, 5-chloropentyl, 6-bromohexyl, 2-thienylmethyl, 1-(2-thienyl)ethyl, 2-(2-thienyl)ethyl, and the like.

[0081] The term "amino acid" refers to any of a class of organic compounds containing at least one amino group, -NH-, and one carboxyl group, -COOH. These compounds can be naturally occurring amino acids present in peptides and can contain any substitution in the amino group, carboxyl group, or side chain. They can also exhibit a different chirality of the naturally occurring amino acids in peptides or have a different backbone, linear, or cyclic, but must exhibit at least one amino group and one carboxyl group, as described above. Amino acids can incorporate functional or protecting groups known to those skilled in the art (T.W. Greene, supra). Suitable amino acids include, but are not limited to, alanine, valine, leucine, and isoleucine.

[0082] Preferably, the organic solvent employed in this synthesis process is selected from methanol, ethanol, propanol, isopropanol, t-butanol, n-butanol, ethyl acetate, isopropyl acetate, butyl acetate, dichloromethane, toluene, tetrahydrofuran, 2-methyltetrahydrofuran, acetonitrile, acetone, cyclopentyl methyl ether, methyl ethyl ketone, methyl isobutyl ketone, dimethylamide, dimethylformamide, and dimethyl sulfoxide.

[0083] As mentioned above, the compounds of formula (I) are useful as calcineurin inhibitors, particularly for use in the prevention and / or treatment of diseases or disorders selected from autoimmune diseases, organ transplant rejection, cancer, and neurological diseases or disorders, preferably for the prevention and / or treatment of autoimmune diseases and / or cancer. Preferably, the autoimmune disease is selected from the group consisting of arthritis, anemia, psoriasis, dermatitis, urticaria, sclerosis, multiple sclerosis, inflammatory bowel disease, Crohn's disease, colitis, endocarditis, endometriosis, episcleritis, respiratory distress syndrome, meningitis, iritis, choroiditis, spondylitis, sudden hearing loss, rhinitis, encephalitis, uveitis, autoimmune myocarditis, leukocyte adhesion deficiency, lupus erythematosus, systemic lupus erythematosus, diabetes, tuberculosis, sarcoidosis, granulomatosis with polyangiitis, vasculitis, autoimmune aplastic anemia, autoimmune neutropenia, CNS inflammatory disorders, Behcet's disease, pemphigoid, pemphigus, Reiter's disease, nephritis, IgM Polyneuropathy, thrombocytopenia, hypothyroidism, hypoparathyroidism, thyroiditis, Graves' disease, autoimmune polyglandular syndrome, paraneoplastic neurological syndrome, encephalomyelitis, myasthenia gravis, autoimmune hepatitis, Guillain-Barré syndrome, Berger's disease, skin diseases, celiac disease, amyotrophic lateral sclerosis, coronary artery disease, autoimmune hearing loss, polychondritis, paraneoplastic neurological syndrome, channelopathies, epilepsy, migraine, autism, fibromyalgia, multiple endocrine neoplasia, presenile dementia, Chagas' disease, rheumatic fever, recurrent miscarriage, erythema multiforme, alveolitis, leprosy, malaria, leishmaniasis, dengue fever, endocarditis, fibrosis , biliary tract inflammation, human immunodeficiency virus (HIV) infection, Alzheimer's disease, Epstein-Barr virus infection, keratoconjunctivitis, bronchitis, chronic obstructive airway disease, idiopathic facial paralysis, chronic fatigue syndrome, rheumatic fever, hypogonadism, pancreatitis, vitiligo, acquired immune deficiency syndrome, multiple organ dysfunction syndrome, autoimmune atrophic gastritis, rheumatic disease, myocarditis, nephrotic syndrome, sinusitis, eosinophilia, hyperalgesia, meningitis, diabetes-related disease, valvulitis, Addison's disease, juvenile dermatomyositis, scleroderma, perioral dermatitis, steroid-resistant nephrotic syndrome, and focal segmental glomerulosclerosis (FSGS).More preferably, the autoimmune disease is selected from arthritis, anemia, psoriasis, dermatitis, sclerosis, multiple sclerosis, meningitis, rhinitis, lupus erythematosus, diabetes, CNS inflammatory disorders, pemphigus, nephritis, autoimmune hepatitis, celiac disease, migraine, fibromyalgia, endocarditis, coronary artery disease, epilepsy, Chagas' disease, rheumatic fever, malaria, Alzheimer's disease, Epstein-Barr virus infection, bronchitis, chronic obstructive airway disease, chronic fatigue syndrome, pancreatitis, acquired immune deficiency syndrome, multiple organ dysfunction syndrome, myocarditis, meningitis, diabetes-related disorders, Addison's disease, juvenile dermatomyositis, scleroderma, perioral dermatitis, steroid-resistant nephrotic syndrome, and focal segmental glomerulosclerosis (FSGS).

[0084] In certain embodiments, the compounds of formula (I) are useful as calcineurin inhibitors, particularly for use in the prevention and / or treatment of cancer, including, but not limited to, Hedgehog signaling pathway-related cancers such as sarcoma, blastoma, nervous system cancer, rhabdomyosarcoma, neuroblastoma, medulloblastoma, intracranial tumors, and brain cancers including brain metastases, glioma, glioblastoma, pediatric brain tumor, embryonal tumor, pineoblastoma, oligodendroglioma, pituitary tumor, Ewing's sarcoma, osteosarcoma, breast cancer, lung cancer, pancreatic cancer, meningioma, ependymoma, craniopharyngioma, choroid plexus papilloma, astrocytic tumor, acoustic neuroma, and soft tissue sarcoma.

[0085] As mentioned above, calcineurin (CN or PPP3, formerly known as PP2B) is a serine / threonine protein phosphatase regulated by Ca2+ and calmodulin. CN regulates development, memory, cardiac function, and immune responses by dephosphorylating various protein substrates, including the cytoplasmic nuclear factor of activated T cells (NFATc) family of transcription factors. NFATc proteins have been shown to be direct substrates of calcineurin. Dephosphorylated NFATc proteins are translocated into the nucleus, where they induce the expression of numerous cytokine and chemokine genes in cooperation with other transcription factors, resulting in T cell activation. It also plays an important role in many physiological processes, including homeostasis, angiogenesis, myogenesis, adipogenesis, osteogenesis, chondrocyte differentiation, circulatory system development, pancreatic beta cell proliferation, hair follicle cell differentiation and remodeling, and activity of cells of the immune and nervous systems, including schizophrenia, diabetes, Down's syndrome, cardiac hypertrophy, and neurodegenerative diseases such as Alzheimer's disease, psychotic disorders, and epilepsy, among others.

[0086] The compounds of formula (I) have surprisingly been found to be inhibitors of the calcineurin-NFAT signaling pathway. NFAT proteins are expressed in immune cells and play a key role in eliciting immune responses. NFAT proteins are activated by calcineurin, and the activated NFAT proteins then induce the transcription of cytokine genes required for immune responses. NFAT or NFATc proteins (nuclear factor of activated T cells) refer to members of a family of transcription factors, including NFAT1, NFAT2, NFAT3, and NFAT4, with several isoforms. Any other NFATc proteins whose activation is dependent on calcineurin are also meant to be included herein. NFATc proteins can be, for example, mammalian proteins, such as human or murine. NFAT1, NFAT2, and NFAT4 are expressed in immune cells, such as T lymphocytes, and play a role in eliciting immune responses. The NFATc protein is involved in the transcriptional regulation of cytokine genes, such as IL-2, IL-3, IL-4, TNF-α, and IFN-γ, during immune responses.

[0087] This action as inhibitors of the calcineurin-NFAT signaling pathway also makes the compounds of formula (I) useful for inhibiting or preventing the activity of the immune system, allowing the compounds of formula (I) to act as immunosuppressants and, surprisingly, in other indications such as cancer.

[0088] Although cancer involves inflammatory processes during its development, its biological complexity and multiple triggering factors make it difficult to classify as an inflammatory disease. Therefore, cancer is a complex disease that goes beyond inflammatory processes. Numerous studies support this assertion. For example, an article by Hanahan and Weinberg titled "The Hallmarks of Cancer" in the 2000 issue of Cell identified six fundamental characteristics of cancer that go beyond inflammation. Furthermore, cancer involves complex genetic and epigenetic changes that go beyond inflammation. While inflammation is an important factor in cancer development, its multifaceted nature and biological diversity distinguish it from purely inflammatory diseases. This assertion is strengthened by the fact that anti-inflammatory agents tested for improving cancer treatment are used as boosters in combination with chemotherapy, the actual anti-cancer treatment, rather than as monotherapy. The present inventors have surprisingly demonstrated that selected compounds of formula (I) exhibit anti-cancer activity in in vitro assays. This finding is particularly relevant considering that the reference compounds used as calcineurin inhibitors (cyclosporin A, tacrolimus, and voclosporin) have not shown anticancer activity and are suspected of being tumorigenic.

[0089] Similarly, while autoimmune diseases are complex conditions characterized by abnormal responses of the immune system against the body's own tissues and cells, inflammation is a common feature in many autoimmune diseases, and they are not classified as inflammatory diseases alone. As noted in a review article by Cooper and Stroehla entitled "The epidemiology of autoimmune diseases" published in Autoimmunity Reviews in 2003, autoimmune diseases encompass a wide range of pathologies with distinct pathogenic mechanisms beyond inflammation, including genetic predisposition and immune dysregulation. Therefore, it is essential to recognize that autoimmune diseases, as distinct from purely inflammatory conditions, include a broader spectrum of immune dysfunction beyond inflammation.

[0090] The present invention therefore relates to immunosuppressants (compounds of formula (I)) that selectively inhibit the interaction between calcineurin and NFAT without inhibiting the enzymatic activity of calcineurin towards other substrates, thereby reducing toxic effects and thus demonstrating reduced side effects compared to gold standard treatments using cyclosporin A or tacrolimus.

[0091] Although the compound of formula (I) potently inhibits NFTAc activation in human HEK293T cells at several concentrations, the compound of formula (I) does not block CN-NFATc interaction through the PxIxIT motif, i.e., the compound of formula (I) behaves as a potential calcineurin inhibitor useful as an immunosuppressant.

[0092] Furthermore, compounds of formula (I) do not affect the phosphatase activity of CNA toward pNPP or Rll substrates, i.e., compounds of formula (I) avoid the serious side effects associated with current immunosuppressants such as cyclosporine or tacrolimus, which bind to calcineurin through the catalytic site.

[0093] In summary, the compounds of formula (I) exhibit more efficient and selective activity as calcineurin inhibitors than current basis drugs such as cyclosporine and tacrolimus.

[0094] In a second aspect, the present invention provides compounds of formula I as defined above, including stereoisomers and pharmaceutically acceptable salts thereof, R1 is

[0095] [ka]

[0096] and R2 is

[0097] [ka]

[0098] and R3, R4, and R6 are independently H, alkyl, halo, or carboxyl; For compounds where R5 is H, halo, or cyano.

[0099] In a preferred embodiment, the compound of formula (I) is R3, R4, and R6 are independently H, C 1~3 alkyl, halo, or carboxyl; R5 is H or halo; At least two of R3, R4, R5, and R6 are compounds different from hydrogen.

[0100] In a preferred embodiment, the compound of formula (I) is R3 is methyl; R4 is H, F, or Cl; R5 is H or F; R6 is H, F, or carboxyl; At least two of R4, R5, and R6 are different from hydrogen; At least one of R4, R5, and R6 is a halogen.

[0101] These new compounds are obtained according to general chemical syntheses found in the literature. An example of a synthetic scheme is shown in Figure 4, in which the compound SLB-605 is obtained. The present invention also includes the use of these new preferred compounds as calcineurin inhibitors, in particular for the prevention and / or treatment of diseases or disorders selected from autoimmune diseases, organ transplant rejection, cancer, and neurological diseases or disorders, preferably for the prevention and / or treatment of autoimmune diseases and / or cancer, including any of the preferred options contained herein.

[0102] The present disclosure also relates to a method for preventing / treating a disease or disorder selected from autoimmune diseases, organ transplant rejection, cancer, and neurological diseases or disorders, preferably a method for preventing and / or treating autoimmune diseases and / or cancer, which, according to embodiments and preferred options contained herein, comprises administering a compound of formula (I) or any of the embodiments, preferably preferred embodiments, contained herein in a therapeutically effective amount, i.e., dosage. The terms "therapeutically effective dosage" and "therapeutically effective amount," in the context of the present invention, refer to an amount of the subject compound that results in prevention, delay of onset of symptoms, or improvement of symptoms. The therapeutically effective amount, as well as the therapeutically effective number of doses, can be determined by conventional methods as discussed below.

[0103] Unless the context dictates otherwise, any of the embodiments disclosed herein can occur alone or in combination with any other embodiment disclosed herein, in other words, for example, preferred options of a stated feature can be combined with more preferred or less preferred options of other features.

[0104] The present invention is illustrated hereinafter by examples which are not intended to limit its scope. Example Example 1A Synthesis of Racemic INK-150.46 (also referred to as SLB-0605)

[0105] [Table 1]

[0106] Step a) tert-Butyl 2-(phenylcarbamoyl)pyrrolidine-1-carboxylate (rac-11) To a stirred solution of racemic Boc-proline (207 mg, 0.962 mmol) in anhydrous CHCl (5 mL) at room temperature, 1-[bis(dimethylamino)methylene]-1H,1,2,3-triazolo[4,5-b]pyridinium 3-hexafluorophosphate oxide (HATU, 439 mg, 1.15 mmol, 1.2 eq) was added. After 10 min, aniline (0.13 mL, 1.44 mmol, 1.5 eq) and triethylamine (0.27 mL, 1.92 mmol, 2 eq) were added dropwise, and the reaction mixture was stirred at room temperature until total consumption of starting material (16 h). The mixture was evaporated to dryness and purified chromatographically on SiO using hexane / EtOAc 93 / 7 to give compound rac-11 (275 mg, 100% purity, 98% yield). 1 H NMR (400MHz, CDCl3) δ9.46(s, 1H), 7.52(d, J=7.6Hz, 2H), 7.31(t, J=7.6Hz, 2H), 7.09(t, J=6.5H) z, 1H), 4.56-4.20(m, 1H), 3.64-3.22(m, 2H), 2.72-2.09(m, 1H), 2.08-1.75(m, 3H), 1.49(s, 9H) HPLC-MS Ret (Method B): 1.79 min; ESI+-MS m / z: 191 (M-Boc)+ Chiral HPLC: Under chiral HPLC conditions, the two enantiomers were successfully separated. ReT (Method Dbis, Column Chiralpak IC): 6.44 min and 9.16 min. Step b) N-phenylpyrrolidine-2-carboxamide (rac-13) To a stirred solution of rac-11 (275 mg, 0.947 mmol) in 6 mL of CHCl was added 2 mL of trifluoroacetic acid (21.8 mmol, 23 eq) at room temperature, and the mixture was sealed and stirred for 0.5 h. The crude product was then evaporated to dryness to give an oily residue of the orange organic salt rac-13, which was used without further purification (quantitative yield). 1H NMR (400MHz, CDCl3) δ11.26(s, 1H), 9.97(s, 1H), 7.52(d, J=7.6Hz, 2H), 7.31-7.26(m, 2H), 7.50-7.26(m, 1H), 7.12(t, J=7.4 Hz, 1H), 5.14-4.74(m, 1H), 3.58-3.39(m, 2H), 2.53(dq, J=7.0, 14.3Hz, 1H), 2.15(dq, J=6.7, 13.0Hz, 1H), 2.09-1.98(m, 2H) HPLC-MS Ret (Method B): 1.23 min; ESI+-MS m / z: 191 (M-Boc)+ Chiral HPLC: Under chiral HPLC conditions, the two enantiomers were successfully separated. ReT (Method Cbis, Column Chiralpak IC): 6.74 min and 8.82 min. Step c) 1-((3-chloro-5-fluoro-2-methylphenyl)sulfonyl)-N-phenylpyrrolidine-2-carboxamide (rac-3) To a stirred solution of rac-13 (288 mg, 0.947 mmol) in anhydrous DMF (4 mL) was added anhydrous EtN (0.46 mL, 3.31 mmol, 3.5 eq) at 0 °C. After 10 min, compound 2 (see Figure 4) was added (345 mg, 1.42 mmol, 1.5 eq) at the same temperature. The mixture was stirred at 0 °C for 2 h and then evaporated to dryness. This was purified on silica gel using hexane / EtOAc from 95 / 5 to 85 / 15 as the eluent to give the desired product rac-3 as a white solid (350 mg, 100% purity, 93% yield). 1 H NMR (400MHz, CDCl3) δ8.60(s, 1H), 7.79(d, J=7.4Hz, 1H), 7.61-7.53(m, 2H), 7.40-7.27(m, 3H), 7.19-7.09(m, 1 H), 4.47-4.31(m, 1H), 3.66-3.54(m, 1H), 3.50-3.36(m, 1H), 2.54-2.43(m, 1H), 2.39(s, 3H), 2.01-1.78(m, 3H) HPLC-MS Ret (Method B): 2.19 min; ESI+-MS m / z: 397(M+H)+ Chiral HPLC: Under chiral HPLC conditions, the two enantiomers were successfully separated. ReT (Method Cbis, Column Chiralpak IC): 6.97 min and 7.67 min Step d) N-((1H-Benzo[d][1,2,3]triazol-1-yl)methyl)-1-((3-chloro-5-fluoro-2-methylphenyl)sulfonyl)-N-phenylpyrrolidine-2-carboxamide (rac-SLB-0605). To a stirred solution of sodium hydride (60% purity, 106 mg, 2.65 mmol, 3 eq) in anhydrous DMF (2 mL) at 0 °C was added a solution of rac-3 (350 mg, 0.882 mmol) in anhydrous DMF (5 mL). After stirring at the same temperature for 1 h, a solution of compound 4 (see Figure 4) (443 mg, 2.65 mmol, 3 eq) in anhydrous DMF (4.5 mL) was added and the mixture was allowed to warm to room temperature overnight. After 16 h, it was evaporated to dryness and purified by silica gel using hexane / EtOAc from 95 / 05 to 70 / 30 as eluent to give the desired product rac-SLB-0605 as a white solid (387 mg, 100% purity, 83% yield). 1 H NMR (400MHz, CDCl3) δ8.05(dt, J=0.9, 8.4Hz, 1H), 7.91(dt, J=0.9, 8.4Hz, 1H), 7.63(d, J=7.4H) z, 1H), 7.53(ddd, J=1.0, 7.0, 8.3Hz, 1H), 7.45-7.31(m, 4H), 7.04(d, J=9.7Hz, 1H), 7.12-6.75( m, 2H), 6.62(d, J=13.8Hz, 1H), 6.37(d, J=13.8Hz, 1H), 4.34(dd, J=4.9, 7.9Hz, 1H), 3.75-3.60( m, 1H), 3.45-3.31(m, 1H), 2.33(s, 3H), 2.17-2.01(m, 1H), 1.88-1.75(m, 2H), 1.75-1.64(m, 1H) HPLC-MS Ret (Method B): 2.33 min; ESI+-MS m / z: 528(M+H)+ Chiral HPLC: The two enantiomers were successfully separated under chiral HPLC conditions. ReT (Method D, Column Chiralpak IA): 11.05 min and 14.08 min Example 1B Synthesis of INK-150.46 (also referred to as SLB-0605)-S enantiomer Step a) Benzyl (S)-4-((tert-butoxycarbonyl)amino)-5-oxo-5-(phenylamino)pentanoate (7, see Figure 4) To a stirred solution of Boc-L-glutamic acid 5-benzyl ester (5 (see Figure 4), 1.007 g, 2.985 mmol) in anhydrous CHCl (15 mL) at 0 °C was added dicyclohexylcarbodiimide (DCC, 739 mg, 3.582 mmol). After 15 min, aniline (0.28 mL, 3.10 mmol) dissolved in anhydrous CHCl (5 mL) was added dropwise to the reaction mixture over 10 min at 0 °C. The orange suspension was stirred at 0 °C for 15 min and then at room temperature until total consumption of the starting material (18 h). The mixture was filtered to remove insoluble material, and the resulting filtrate was evaporated to dryness and purified chromatographically on SiO using hexane / ETOAc 85 / 15 to give compound 7 (see Figure 4) (1.231 g, 100% purity, 100% yield). 1 H NMR (400MHz, CDCl3) δ8.35(s, 1H), 7.50(dd, J=1.0, 8.6Hz, 2H), 7.40-7.27(m, 7H), 7.11(t, J=7.4Hz, 1H), 5.33(d, J=7.5Hz, 1H), 5.15(d, J=2.5Hz, 2H) ), 4.38-4.17(m, 1H), 2.65(ddd, J=6.4, 7.7, 16.8Hz, 1H), 2.51(dt, J=6.7, 17.0Hz, 1H), 2.33-2.15(m, 1H), 2.01(dq, J=6.8, 14.4Hz, 1H), 1.45(s, 9H) HPLC-MS Ret (Method B): 2.25 min; ESI+-MS m / z: 435(M+Na)+ Chiral HPLC: This compound was subjected to different chiral HPLC conditions and in all cases only one peak was observed. Ret (Method C, Column Chiralpak AS-H): 5.86 min Step b) tert-butyl (S)-(5-hydroxy-1-oxo-1-(phenylamino)pentan-2-yl)carbamate (8, see Figure 4) To a stirred suspension of NaBH (448 mg, 11.83 mmol) in EtOH (0.14 L) at 0 °C, ground CaCl (689 mg, 6.21 mmol) was carefully added over 1 min. Compound 7 (see Figure 4) (1.220 g, 2.958 mmol) dissolved in EtOH (20 mL + 5 mL + 5 mL) was then added portionwise over 10 min. The solution was stirred for 3.5 h and allowed to warm to room temperature. The crude product was neutralized with saturated aqueous NH Cl at 0 °C, and the aqueous phase was extracted with EtOAc. The organic phase was washed with saturated aqueous NaCl, dried over anhydrous Na SO , and evaporated to dryness. The resulting oily residue was purified by chromatography on SiO using hexane / ETOAc (35:65) to give compound 8 (see Figure 4) (909 mg, 100%). 1 H NMR (400MHz, CDCl3)δ 8.67(s, 1H), 7.51(dd, J=1.1, 8.6Hz, 2H), 7.37-7.27(m, 2H), 7.13-7.05(m, 1H), 5.42(d, J=7.2Hz, 1H), 4.49-4.29(m, 1H), 3.87-3.70(m, 2H), 2.42(s, 1H), 1.99(dt, J=6.9, 13.7Hz, 1H), 1.84(dq, J=7.4, 13.9Hz, 1H), 1.74-1.63(m, 2H), 1.45(s, 9H) HPLC-MS Ret (Method B): 1.52 min; ESI+-MS m / z: 331(M+Na)+ Chiral HPLC: This compound was subjected to different chiral HPLC conditions and in all cases only one peak was observed. Ret (Method D, Column Chiralpak IB): 5.34 min Step c) (S)-4-((tert-butoxycarbonyl)amino)-5-oxo-5-(phenylamino)pentyl methanesulfonate (9, see Figure 4) To a stirred solution of compound 8 (see Figure 4) (0.898 g, 2.912 mmol) in anhydrous CHCl (30 mL) at 0 °C was added anhydrous EtN (0.61 mL, 4.37 mmol, 1.5 eq). After 10 min, methanesulfonyl chloride (0.27 mL, 3.49 mmol, 1.2 eq) was added dropwise at 0 °C. The mixture was stirred at 0 °C for 10 min and at room temperature for 2.5 h. The crude product was then purified by chromatography on SiO using hexane / ETOAc (50:50) to give 9 (see Figure 4) (1.111 g, 99% yield), which was used in the following step. 1H NMR (400MHz, CDCl3) δ8.23(s, 1H), 7.52(dd, J=1.1, 8.6Hz, 2H), 7.37-7.28(m, 2H), 7.11(t, J=7.4Hz, 1H), 5.21(d, J=5.5Hz, 1H), 4.56-4.42 (m, 1H), 4.41-4.26(m, 1H), 4.31(dt, J=5.1, 10.3Hz, 1H), 3.05(s, 3H), 2.16-2.05(m, 1H), 2.00-1.84(m, 2H), 1.84-1.71(m, 1H), 1.46(s, 9H) HPLC-MS Ret (Method B): 1.80 min; ESI+-MS m / z: 409(M+Na)+ Chiral HPLC: This compound was subjected to different chiral HPLC conditions and in all cases only one peak was observed. Ret (Method D, Column Chiralpak AD-H): 18.37 min Step d) (S)-tert-butyl 2-(phenylcarbamoyl)pyrrolidine-1-carboxylate (11, see Figure 4) To a stirred solution of 9 (see Figure 4) (1.109 g, 2.87 mmol) in anhydrous tetrahydrofuran (24 mL) at 50 °C under an argon atmosphere, potassium tert-butoxide (0.34 g, 3.01 mmol, 1.05 eq) was added and the mixture was continued stirring for 1 h at 50 °C. The crude reaction was evaporated to dryness and the residue was purified chromatographically on SiO using hexane / ETOAc (85:15) to give 11 (see Figure 4) (405 mg, 49% yield). 1H NMR (400MHz, CDCl3) δ7.56 (d, J=7.7Hz, 2H), 7.32 (t, J=7.5Hz, 2H), 7.10 (t, J=6.8Hz, 1H), 4.47-4.20 (m, 1H) ), 3.71-3.38(m, 2H), 2.41-2.17(m, 1H), 2.03(dq, J=5.8, 6.3, 11.2Hz, 2H), 1.97-1.77(m, 1H), 1.37(s, 9H) HPLC-MS Ret (Method B): 1.79 min; ESI+-MS m / z: 313(M+Na)+ Chiral HPLC: This compound was subjected to the same chiral HPLC conditions used in the racemic route for this intermediate, and peaks corresponding to the two enantiomers were observed in a ratio of 94.6:5.4 (89.2% ee). Ret (Method Dbis, Column Chiralpak IC): 6.45 min and 9.20 min, ratio 94.6:5.4 (89.2%ee) Upon increasing the chromatographic polarity (from 25 to 40% EtOAc), the six-membered isomer (10, see Figure 4) was also separated (230 mg, 28% yield). Under standard HPLC conditions (Method B), both compounds 10 and 11 (see Figure 4) eluted very closely, and the separation was excellent. 1 H NMR (400MHz, CDCl3) δ7.43-7.35(m, 2H), 7.29-7.20(m, 3H), 5.54(s, 1H), 4.26(dt, J=5.6, 11.3Hz, 1H), 3.70(td , J=1.7, 6.4Hz, 2H), 2.61(dq, J=5.3, 11.3Hz, 1H), 2.16-1.98(m, 2H), 1.71(tt, J=8.4, 12.3Hz, 1H), 1.46(s, 9H) HPLC-MS Ret (Method B): 1.75 min; ESI+-MS m / z: 291(M+H)+ Step e) (S)—N-phenylpyrrolidine-2-carboxamide (trifluoroacetate salt, 13, see FIG. 4) To a stirred solution of 11 (see Figure 4) (395 mg, 1.36 mmol) in CHCl (10 mL) was added trifluoroacetic acid (2.4 mL, 31.29 mmol, 23 eq) at room temperature, and the mixture was sealed and stirred for 1 h. The crude product was evaporated to dryness to give an orange oily residue of organic salt 13 (see Figure 4), which was used without further purification (quantitative yield). 1 H NMR (400MHz, CDCl3) δ11.54(s, 1H), 10.06(s, 1H), 7.48(dd, J=1.1, 8.6Hz, 2H), 7.45-7.29(m, 1H), 7.30-7.24(m, 2H), 7.18-7. 02(m, 1H), 4.87(t, J=7.5Hz, 1H), 3.49-3.30(m, 2H), 2.60-2.43(m, 1H), 2.14(td, J=6.7, 13.0, 13.6Hz, 1H), 2.08-1.93(m, 2H) HPLC-MS Ret (Method B): 1.23 min; ESI+-MS m / z: 191(M+H)+ Chiral HPLC: This compound was subjected to the same chiral HPLC conditions used in the racemic route for this intermediate, and peaks corresponding to the two enantiomers were observed in a ratio of 94.1:5.9 (88.2% ee). Ret (Method Cbis, Column Chiralpak IC): 6.74 min and 8.83 min, ratio 94.1:5.9 (88.2%ee) Step f) (S)-1-((3-chloro-5-fluoro-2-methylphenyl)sulfonyl)-N-phenylpyrrolidine-2-carboxamide (3, see Figure 4) To a stirred solution of 13 (see Figure 4) (389.8 g, 1.28 mmol) in anhydrous DMF (5 mL) was added anhydrous EtN (0.62 mL, 4.48 mmol, 3.5 eq) at 0 °C. After 10 min, compound 2 (see Figure 4) was added (467 mg, 1.92 mmol, 1.5 eq) at 0 °C, and the mixture was stirred at the same temperature for 2 h. Then, the solvent was removed under vacuum, and the residue was purified by silica gel using hexane / ETOAc 95 / 5 to 85 / 15 as eluent to give the desired product 3 as a white solid (508 mg, 100% purity, 100% yield) (see Figure 4). 1 H NMR (400MHz, CDCl3) δ8.60(s, 1H), 7.79(dd, J=3.5, 7.3Hz, 1H), 7.64-7.53(m, 2H), 7.40-7.27(m, 3H), 7.18-7.09( m, 1H), 4.44-4.33(m, 1H), 3.65-3.52(m, 1H), 3.48-3.37(m, 1H), 2.53-2.42(m, 1H), 2.39(s, 3H), 2.05-1.79(m, 3H) HPLC-MS Ret (Method B): 2.19 min; ESI+-MS m / z: 397(M+H)+ Chiral HPLC: This compound was subjected to the same chiral HPLC conditions used in the racemic route for this intermediate, and peaks corresponding to the two enantiomers were observed in a ratio of 5.9:94.1 (88.2% ee). Ret (Method Cbis, Column Chiralpak IC): 6.98 min and 7.98 min, ratio 5.9:94.1 (88.2%ee) Step g) (S)—N-((1H-benzo[d][1,2,3]triazol-1-yl)methyl)-1-((3-chloro-5-fluoro-2-methylphenyl)sulfonyl)-N-phenylpyrrolidine-2-carboxamide (SLB-0605) To a stirred solution of sodium hydride (154 mg, 3.84 mmol) in anhydrous DMF (2.5 mL) at 0 °C under an argon atmosphere was added a solution of 3 (see Figure 4) (508 mg, 1.28 mmol) in anhydrous DMF (7 mL). After 1 h at 0 °C, anhydrous DMF (5 After addition of 1-(chloromethyl)-1H-benzotriazole (644 mg, 3.84 mmol, 3 eq) in 1 mL of HCl, the reaction mixture was allowed to reach room temperature and stirred for 18 h. Afterwards, the solvent was removed under vacuum and purified on silica gel using hexane / ETOAc 95 / 5 to 75 / 25 as eluent to give the desired product SLB-0605 as a white solid (604 mg, 100% purity, 89% yield). 1 H NMR(400 MHz, CDCl3)δ8.05(dt, J=0.9, 8.4Hz, 1H), 7.91(dt, J=0.8, 8.4Hz, 1H), 7.63(d, J=7.5Hz, 1H), 7.53(d dd, J=1.0, 7.0, 8.2Hz, 1H), 7.47-7.32(m, 4H), 7.04(d, J=9.7Hz, 1H), 7.11-6.77(m, 2H), 6.62(d, J=1 3.8Hz, 1H), 6.37(d, J=13.8Hz, 1H), 4.35(dd, J=4.8, 7.8Hz, 1H), 3.67(dt, J=7.1, 9.4Hz, 1H), 3.38(d t, J=6.7, 9.3Hz, 1H), 2.33(s, 3H), 2.18-2.05(m, 1H), 1.84-1.74(m, 2H), 1.69(dt, J=6.6, 12.3Hz, 1H) HPLC-MS Ret (Method B): 2.33 min; ESI+-MS m / z: 528(M+H)+ Chiral HPLC: This compound was subjected to the same chiral HPLC conditions used in the racemic route for this final compound, and two peaks were observed in a ratio of 5.9:94.1 (88.2% ee). Ret (Method D, Column Chiralpak IA): 11.07 minutes and 14.10 minutes, ratio 5.9:94.1 (88.2%ee) Analysis method: Method A (not adopted): Column ZORBAX Extend-C18 RRHD2.1×50mm, 1.8μm, temperature 35℃; flow rate 0.61mL / min; A:NH4HCO3 10 mM B: MeCN; gradient 0.3 min 98% A to 2.65 min 98% A to 100% B; isocratic 2.05 min 100% B Method B: Column: ZORBAX Extend-C18 RRHD 2.1 x 50 mm, 1.8 μm, temperature: 35 °C; flow rate: 0.61 mL / min; A: 10 mM NH₄HCO₃, B: MeCN, C: MeOH + 0.1% formic acid; gradient: 0.3 min 98% A, 2.7 min 98% A to 0:95:5 A:B:C; 0.1 min 0:95:5 A:B:C to 100% B; isocratic: 2 min 100% B Method C (not adopted): Column ZORBAX Extend-C18 RRHD4.6mm×250mm, 5μm, temperature: room temperature; eluent n-Heptane / EtOH / DEA50 / 50 / 0.1 v / v / v, flow rate: 0.8ml / min Method Cbis:Column ZORBAX Extend-C18 RRHD4.6mm×250mm, 5μm, temperature: room temperature; eluent n-Heptane / EtOH / DEA70 / 30 / 0.1 / v / v, flow rate: 0.8ml / min Method D: Column ZORBAX Extend-C18 RRHD4.6mm×250mm, 5μm, temperature: room temperature; eluent n-Heptane / IPA / DEA50 / 50 / 0.1 v / v / v, flow rate: 0.8ml / min Method Dbis:Column ZORBAX Extend-C18 RRHD4.6mm×250mm, 5μm, Temperature: Room temperature; Eluent n-Heptane / IPA / DEA70 / 30 / 0.1 v / v / v, Flow rate: 0.8ml / min Example 2 Compounds of formula (I) inhibit NFATc activity in HEK293T cells To assess the effect of compounds of Formula (I) on NFATc activity, HEK293T cells were transfected with the 3xNFAT-luc reporter gene and Renilla-luc plasmid as a control for gene transfection. After 24 hours, cells were stimulated with ionomycin (Io), PMA, and CaCl2 for 4 hours to elevate intracellular calcium levels and activate the cellular CN-NFATc signaling pathway. As a positive control for the inhibition of CN activity, cells were treated with 1 μM CsA 30 minutes before Io / PMA / CaCl2 stimulation. Cells were then treated with compounds of Formula (I) at several concentrations for 2 hours and analyzed using a dual-luciferase reporter assay system at a light unit according to the manufacturer's protocol (Figure 1).

[0107] The results in Figure 1 demonstrate that compounds INK-150.35, INK-150.46, and INK-150.48 potently inhibit NFATc activation in HEK293T cells, occurring at compound concentrations of at least 100 μM, 50 μM, and 20 μM.

[0108] [Table 2]

[0109] Example 3 Compounds of formula (I) do not inhibit calcineurin phosphatase activity towards pNPP substrate CN dephosphorylation activity is typically assessed against the small molecule pNPP to determine the accessibility of the small molecule to the active site of the CN enzyme, and against the RII peptide from the regulatory RII subunit of cAMP-dependent protein kinase as a large substrate that binds to CNA prior to dephosphorylation by the CN active site.

[0110] To determine the functional effects of compounds of formula (I) on CN phosphatase activity toward pNPP substrate, a biochemical assay was performed. The assay evaluates the ability of recombinant human CN (amino acids 2-347) conjugated to glutathione S-transferase (GST-CN) to dephosphorylate pNPP substrate. First, GST-CN was expressed in large quantities in bacteria, purified, and then CN activity toward pNPP substrate was analyzed in the absence of any compound. Next, 50 μM of each compound was analyzed to determine their potential inhibitory ability toward CN phosphatase activity (Figure 2). Example 4 Compounds of formula (I) do not inhibit calcineurin phosphatase activity towards RII substrates The compound INK-150.00, which inhibits NFATc activity, was also analyzed for its ability to inhibit CN full-length protein against the RII substrate using a CN phosphatase assay kit (Enzo Life Sciences) according to the manufacturer's instructions.

[0111] Compounds of formula (I) do not appear to affect the phosphatase activity of CN towards RII substrates (Figure 3).

[0112] In conclusion, the compound of formula (I) does not affect the phosphatase activity of CN against pNPP or RII substrates. The compound of formula (I) shows potent inhibitory effects on NFATc activity in human 293T cells at several concentrations.

[0113] However, the compound of formula (I) competes for NFATc binding to CN through a site different from the aforementioned PxlxlT and LxVP binding motifs in CN. Example 5 Assay of compound SLB-605 against several cancer cell lines The compound INK-150.46 (ie, SLB-605) was analyzed against several cancer cell lines in proliferation assays.

[0114] All measurements, except for D283 cells, were performed using an in vitro assay to measure cell growth based on methyl violet staining after 5 days of incubation with the above-mentioned drugs or control vehicle (DMSO). The procedure was performed in 96-well plates. After one wash with phosphate-buffered saline (PBS), cells were stained with methyl violet (0.5% w / v in water) for 20 minutes at room temperature. After five washes with PBS and the addition of acetic acid (10% in water), plates were read in a spectrophotometer at 590 nm. All conditions were tested in six independent wells, and results were expressed as the average of all replicates.

[0115] The D283 cell assay was performed in a 12-well plate. Cells were seeded at low density and cultured in EMEM medium for 24 hours. The floating cells in each well were collected by centrifugation of the recovered medium, and the corresponding treatment was administered and returned to the well. After 5 days of treatment, all cells recovered and were directly counted using an EVE® automated cell counter (NanoEntek).

[0116] As can be seen, compound SLB-605 showed good inhibition of cell culture in the strains assayed.

[0117] See Figures 5, 6, 7, 8 and the table below.

[0118] [Table 3]

[0119] The table above shows the inhibitory effects of compounds on several cancer cell lines. The results are expressed as % inhibition, with higher values ​​indicating better results. Figure 5 visually illustrates the inhibitory effect of compound SLB-605 on two different rhabdomyosarcoma (RMS) cell lines, RD and RH4. Dimethyl sulfoxide (DMSO) was used as a control. N1 and n2 represent two independent experiments. Results are expressed as % proliferation values, with smaller values ​​indicating better results.

[0120] Figure 6 visually illustrates the inhibitory effect of compound SLB-605 on two different neuroblastoma (NB) cell lines, SKNBE2 and SHSY5Y. Dimethyl sulfoxide (DMSO) was used as a control. N1 and n2 represent two independent experiments. Results are expressed as % proliferation values, with smaller values ​​indicating better results.

[0121] Figure 7 visually illustrates the inhibitory effect of compound SLB-605 on the osteosarcoma (OS) cell line U2OS. Dimethyl sulfoxide (DMSO) was used as a control. Results are expressed as % proliferation values, with lower values ​​indicating better results.

[0122] Figure 8 visually illustrates the inhibitory effect of compound SLB-605 on the medulloblastoma (MB) cell line D283. Dimethyl sulfoxide (DMSO) was used as a control. N1 and n2 represent two independent experiments. Results are expressed as % proliferation values, with smaller values ​​indicating better results.

Claims

1. 1. A compound for use in the prevention and / or treatment of a disease or disorder selected from autoimmune diseases, organ transplant rejection, cancer, and neurological diseases and disorders, comprising: Formula I 【Chemical 1】 and represented by formula I, including stereoisomers and pharmaceutically acceptable salts thereof, R １ is hydrogen; alkyl optionally substituted with Het; benzoyl optionally substituted with alkyl or cyano; or Het, R ２ is aryl optionally substituted with 1, 2, or 3 substituents each independently selected from alkyl, halo, cyano, amido, and carboxyl; benzyl optionally substituted with 1, 2, or 3 substituents each independently selected from alkyl, halo, cyano, amido, and carboxyl; Each Het as a group or part of a group is a monocyclic ring of 5 or 6 atoms; or a bicyclic ring structure consisting of a 6-membered ring fused to a 4-, 5-, or 6-membered ring; each of said rings is saturated, partially unsaturated, or fully unsaturated, and at least one of said rings contains 1 to 4 heteroatoms each independently selected from nitrogen, oxygen, and sulfur, and any of said rings is selected from halo, hydroxyl, nitro, cyano, C １～６ Alkyl, and C １～６ The compound optionally substituted with one, two, or three substituents each independently selected from the group consisting of alkoxy.

2. R １ is hydrogen; C optionally substituted with Het １～６ Alkyl; C １～６ benzoyl optionally substituted with alkyl or cyano; or Het, R ２ is C １～６ aryl optionally substituted with 1, 2, or 3 substituents each independently selected from alkyl, halo, cyano, amido, and carboxyl; or C １～６ benzyl optionally substituted with 1, 2, or 3 substituents each independently selected from alkyl, halo, cyano, and carboxyl; each Het as a group or part of a group is a monocyclic ring with 5 or 6 atoms; or a bicyclic ring structure consisting of a 6-membered ring fused to a 5-membered ring or a 6-membered ring, each of which is saturated, partially unsaturated, or fully unsaturated, and at least one of the rings contains 1 to 4 heteroatoms each independently selected from nitrogen, oxygen, and sulfur, and any of the rings is selected from halo, hydroxyl, cyano, C １～６ 10. The compound for use according to claim 1, optionally substituted with 1, 2, or 3 substituents each independently selected from the group consisting of alkyl.

3. R １ is hydrogen; methyl optionally substituted with Het; or benzoyl optionally substituted with cyano, R ２ is aryl optionally substituted with one, two, or three substituents each independently selected from methyl, halo, cyano, amido, and carboxyl; or benzyl optionally substituted with one, two, or three substituents each independently selected from methyl, halo, and cyano; 3. The compound for use according to any one of claims 1 and 2, wherein each Het as a group or part of a group is a bicyclic ring structure consisting of a 6-membered ring fused to a 5-membered ring or a 6-membered ring, each of said rings being partially unsaturated, and at least one of said rings containing 1 to 4 heteroatoms each independently selected from nitrogen, oxygen, and sulfur.

4. R １ teeth, 【Chemistry 2】 【Chemistry 3】 【Chemistry 4】 【Chemistry 5】 and 【Chemistry 6】 Selected from R ２ teeth, 【Chemistry 7】 【Chemistry 8】 【Chemistry 9】 【Chemistry 10】 【Chemistry 11】 【Chemistry 12】 【Chemistry 13】 【Chemistry 14】 【Chemistry 15】 【Chemistry 16】 【Chemistry 17】 and 【Chemistry 18】 The compound according to any one of claims 1 to 3, selected from the group consisting of:

5. R １ teeth, 【Chemistry 19】 and R ２ teeth, 【Chemistry 20】 The compound according to any one of claims 1 to 3, wherein

6. The compound for use according to any one of claims 1 to 5, wherein the disease or disorder is cancer.

7. The compound for use according to any one of claims 1 to 5, wherein the disease or disorder is an autoimmune disease.

8. The autoimmune diseases include arthritis, anemia, psoriasis, dermatitis, urticaria, sclerosis, multiple sclerosis, inflammatory bowel disease, Crohn's disease, colitis, endocarditis, endometriosis, episcleritis, respiratory distress syndrome, meningitis, iritis, choroiditis, spondylitis, sudden hearing loss, rhinitis, encephalitis, uveitis, autoimmune myocarditis, leukocyte adhesion deficiency, lupus erythematosus, systemic lupus erythematosus, diabetes, tuberculosis, sarcoidosis, granulomatosis with polyangiitis, vasculitis, and autoimmune aplastic anemia. , autoimmune neutropenia, CNS inflammatory disorders, Behçet's disease, pemphigoid, pemphigus, Reiter's disease, nephritis, IgM polyneuropathy, thrombocytopenia, hypothyroidism, hypoparathyroidism, thyroiditis, Graves' disease, polyglandular autoimmune syndrome, paraneoplastic neurological syndrome, encephalomyelitis, myasthenia gravis, autoimmune hepatitis, Guillain-Barré syndrome, Berger's disease, skin disorders, celiac disease, amyotrophic lateral sclerosis, coronary artery disease, autoimmune hearing loss, multiple myeloma Osteitis, paraneoplastic neurological syndrome, channelopathy, epilepsy, migraine, autism, fibromyalgia, multiple endocrine neoplasia, presenile dementia, Chagas disease, rheumatic fever, recurrent miscarriage, erythema multiforme, alveolitis, leprosy, malaria, leishmaniasis, dengue fever, endocarditis, fibrosis, cyclitis, human immunodeficiency virus (HIV) infection, Alzheimer's disease, Epstein-Barr virus infection, keratoconjunctivitis, bronchitis, chronic obstructive airway disease, idiopathic facial paralysis, chronic 8. The compound for use according to claim 7, selected from fatigue syndrome, rheumatic fever, hypogonadism, pancreatitis, vitiligo, acquired immune deficiency syndrome, multiple organ dysfunction syndrome, autoimmune atrophic gastritis, rheumatic diseases, myocarditis, nephrotic syndrome, sinusitis, eosinophilia, hyperalgesia, meningitis, diabetes-related diseases, valvulitis, Addison's disease, juvenile dermatomyositis, scleroderma, perioral dermatitis, steroid-resistant nephrotic syndrome, and focal segmental glomerulosclerosis (FSGS).

9. 9. The compound for use according to claim 8, wherein the autoimmune disease is selected from arthritis, anemia, psoriasis, dermatitis, sclerosis, multiple sclerosis, meningitis, rhinitis, lupus erythematosus, diabetes, CNS inflammatory disorders, pemphigus, nephritis, autoimmune hepatitis, celiac disease, migraine, fibromyalgia, endocarditis, coronary artery disease, epilepsy, Chagas' disease, rheumatic fever, malaria, Alzheimer's disease, Epstein-Barr virus infection, bronchitis, chronic obstructive airway disease, chronic fatigue syndrome, pancreatitis, acquired immune deficiency syndrome, multiple organ dysfunction syndrome, myocarditis, meningitis, diabetes-related diseases, Addison's disease, juvenile dermatomyositis, scleroderma, perioral dermatitis, steroid-resistant nephrotic syndrome, and focal segmental glomerulosclerosis (FSGS).

10. 7. The compound for use according to claim 6, wherein the cancer comprises sarcoma, blastoma, nervous system cancer, rhabdomyosarcoma, neuroblastoma, medulloblastoma, intracranial tumors and brain metastases, brain cancer including glioma, glioblastoma, pediatric brain tumor, embryonal tumor, pineoblastoma, oligodendroglioma, pituitary tumor, Ewing's sarcoma, osteosarcoma, breast cancer, lung cancer, pancreatic cancer, meningioma, ependymoma, craniopharyngioma, choroid plexus papilloma, astrocytic tumor, acoustic neuroma, and soft tissue sarcoma.

11. A compound of formula I, R １ teeth, 【Chemical 21】 and R ２ teeth, 【Chemical 22】 and R ３ , R ４ , and R ６ are independently H, alkyl, halo, or carboxyl; R ５ is H, halo, or cyano.

12. R ３ , R ４ , and R ６ are independently H, C １～３ alkyl, halo, or carboxyl; R ５ is H or halo, R ３ , R ４ , R ５ , and R ６ 12. The compound of claim 11, wherein at least two of are different from hydrogen.

13. R ３ is methyl, R ４ is H, F, or Cl, R ５ is H or F, R ６ is H, F, or carboxyl; R ４ , R ５ , and R ６ At least two of these are different from hydrogen, R ４ , R ５ , and R ６ 13. The compound according to any one of claims 11 and 12, wherein at least one of is halogen.

14. R ３ is methyl, R ４ is Cl, R ５ is H, R ６ The compound according to any one of claims 11 to 13, wherein

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