EA045073B1 - CHROMENE DERIVATIVES AS TCR-NCK INTERACTION INHIBITORS - Google Patents

Description

Field of technology to which the invention relates

The present invention relates to a group of compounds containing a chromene core and having the ability to inhibit lymphocyte proliferation by blocking the interaction of TCR with Nck. Thus, such compounds can be used to treat diseases, disorders or conditions in which such interaction contributes to complications such as transplant rejection reactions, immune or autoimmune disease or proliferation.

State of the art

Autoimmune and inflammatory diseases, such as asthma, multiple sclerosis (MS), allergies, rheumatoid arthritis (RA), Crohn's disease or psoriasis, are a group of heterogeneous diseases in which the adaptive immune system, particularly through T lymphocytes, attacks self-antigens body. It is generally accepted that T cells are at the center of all immunological mechanisms. T cells can recognize both foreign and self antigens and activate an immune response against them. T cells recognize antigens through the T cell antigen receptor (TCR), which is responsible for transmitting the signal to the cytoplasm. Indeed, the fact that the major histocompatibility complex (MHC) haplotype is the most important genetic risk factor for human autoimmune disease places T cells at the center of all immunopathological phenomena.

The T cell recognizes MHC-related antigen peptide (pMHC) through the TCR and is capable of translating small changes in pMHC chemistry into various quantitative and qualitative outputs. Although a variety of control mechanisms exist to prevent the activation of T cells bearing TCRs with significant affinity for MHC loaded with self-peptides, including suppression of potentially autoreactive T cells during their maturation in the thymus, these mechanisms are somewhat insufficient for patients who develop autoimmune diseases, and autoreactive T cells are activated and proliferate, overcoming homeostatic control mechanisms.

Upon stimulation, the TCR is activated and undergoes a conformational change, which leads to the recruitment of various proteins that form the TCR signalosome, responsible for signal transduction and cell activation. This complex includes the cytosolic protein non-catalytic region of the protein tyrosine kinase (Nck), which binds to the proline-rich sequence (PRS) motif present in the CD3ε subunit of the TCR. As a result, this TCR conformational change is stabilized and the activation signal is effectively transmitted.

Current therapies for immune diseases are immunosuppressive strategies distinct from tolerogenic/immunomodulatory approaches. Azathioprine, methotrexate, mycophenolate and cladribine are cytostatics. Other therapies cause T cell depletion (alemtuzumab, anti-CD25) or retention in lymph nodes (Fingolimod). Alternatively, indirect modulation of the immune system (BG-12) is also used as a strong strategy. Thus, despite the central role of the TCR signal for T cell activation in autoimmune diseases, recent attempts to modulate T cell activation have focused on modulating co-stimulatory signals, cytokine receptors, etc., but they have the corresponding disadvantage of specificity and have a large number of associated side effects.

To develop specific immunomodulatory therapies, many different groups of researchers have attempted to determine the role of Nck in T cell activation. Nck has been attributed an important role in the function of mature T cells based on studies in knockout mice, which lack Nck1 in all tissues and lack Nck2 under certain conditions only in T cells. In these models, the number of peripheral T cells expressing TCRs with low avidity for self-antigens was dramatically reduced and there was a general deterioration in T cell activation when stimulated with weak antigens. Moreover, the importance of Nck has also been addressed by generating bone marrow chimeras, in which the PRS motif (Nck binding site in the TCR) was shown to be important for the activation of mature T cells by weak agonists, but not by strong ones. Similarly, mutation of the PRS motif altered the ability of mice to activate the adaptive immune response in vivo. In addition, a high-affinity inhibitory peptide to the SH3.1 domain of Nck alters TCR signalosome assembly, suggesting that Nck recruitment is a critical early step in TCR signaling that is targeted for modulation of the immune system.

WO 2010/064707 describes a series of compounds derived from 2H-chromene for the prevention or treatment of disease caused by sphingosine-1-phosphate (S1P1)-mediated unwanted lymphocyte infiltration. WO 2012/042078 also describes chromene derivatives having inhibitory properties against the TCR-Nck interaction in T cells and their use for the treatment of autoimmune diseases, inflammatory diseases or transplant rejection.

Thus, it is desirable to obtain new compounds that have the ability to inhibit TCR-Nck interactions in T lymphocytes and are suitable drug candidates. Compounds must exhibit good activity in in vivo pharmacological studies, good oral absorption, and be metabolically stable and exhibit a favorable pharmacokinetic profile. In addition, such compounds should not be toxic and should provide minimal side effects.

Brief description of the invention

In the present invention, it has been found that the compounds of the present invention and pharmaceutically acceptable compositions thereof are effective as modulators of the interaction of TRC with Nck. Such compounds have the general formula I:

or is a pharmaceutically acceptable salt of the specified compound, where each variable is as defined and described herein. The compounds of the present invention and pharmaceutically acceptable compositions thereof are suitable for use in the treatment of a variety of diseases, disorders or conditions associated with T cell activation. Such diseases, disorders or conditions include the diseases, disorders or conditions described herein.

Detailed description of some implementation options

General Description of Some Embodiments of the Present Invention: Some embodiments of the present invention provide a compound of formula I:

I or a pharmaceutically acceptable salt thereof, wherein: R ¹ is R, halogen, -CN, -OR or -N(R)2; R ² is R, halogen, -C(O)N(R)2 or -N(R)2; R ³ represents hydrogen or an electron-withdrawing group;

each R independently represents hydrogen or an optionally substituted group selected from a C _1-6 aliphatic group, a 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring, phenyl, a 4-8 membered saturated or partially unsaturated monocyclic heterocyclic ring containing 1 -2 heteroatoms independently selected from nitrogen, oxygen or sulfur, a 5-6 membered monocyclic heteroaromatic ring containing 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur; or two R groups on the same nitrogen atom are combined with intervening atoms to form an optionally substituted 5-6 membered heterocyclic ring containing 0-1 heteroatom in addition to the nitrogen atom attached to said ring, such heteroatom being oxygen , nitrogen or sulfur;

L ¹ represents a covalent bond or C _1-4 bivalent straight or branched saturated or unsaturated hydrocarbon chain in which 1-2 methylene chain units are independently and optionally replaced by -O-, -C(O)-, -C(O) O-, -OC(O)-, -C(S)-, -C(R)2-, -C(F)2-, -N(R)-, -C(O)N(R)- , -RNC(O)-, -OC(O)N(R)-, -N(R)C(O)N(R) - or -Су-;

L ² represents a covalent bond or C _1-4 bivalent straight or branched saturated or unsaturated hydrocarbon chain in which 1-2 methylene chain units are independently and optionally replaced by -O-, -C(O)-, -C(O) O-, -OC(O)-, -C(S)-, -C(R) ₂ -, -CH(R)-, -C(F)2-, -N(R)-, -C( O)N(R)-, -RNC(O)-, -OC(O)N(R)- or -N(R)C(O)N(R)-; And

Cy is a bivalent optionally substituted 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring, optionally substituted phenylene, optionally substituted 4-8 membered saturated or partially unsaturated monocyclic heterocyclic ring containing 1-3 heteroatoms independently selected from nitrogen, oxygen or sulfur, an optionally substituted 5-6 membered monocyclic heteroaromatic ring containing 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

Some embodiments of the present invention provide a compound of formula I, wherein said compound is not:

- 2 045073

- 3 045073

Some embodiments of the present invention provide a compound of formula I, wherein said compound is not:

- 4 045073

- 5 045073

52

Connections and definitions.

The compounds of the present invention include those generally described herein and further illustrated by the classes, subclasses and types disclosed herein. Unless otherwise specified, the following definitions shall apply throughout this document. For the purposes of the present invention, the chemical elements are identified in accordance with the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, ^75th Ed. In addition, the general principles of organic chemistry are described in Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito: 1999, and March's Advanced Organic Chemistry, ^5th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons , New York: 2001, the entire contents of said references are incorporated herein by reference. The term aliphatic or aliphatic group as used herein means a straight-chain (i.e., straight-chain) or branched substituted or unsubstituted hydrocarbon chain that is fully saturated or that contains one or more units of unsaturation, or a monocyclic hydrocarbon or bicyclic hydrocarbon that is fully saturated or which contains one or more unsaturation units but is not aromatic (also referred to herein as carbocycle, cycloaliphatic or cycloalkyl) in which or which has one attachment site to the rest of the molecule. Unless otherwise specified, aliphatic groups contain 1-6 aliphatic carbon atoms. In some embodiments, the aliphatic groups contain 1-5 aliphatic carbon atoms. In other embodiments, the aliphatic groups contain 1-4 aliphatic carbon atoms. In other embodiments, the aliphatic groups contain 1-3 aliphatic carbon atoms, and in other embodiments, the aliphatic groups contain 1-2 aliphatic carbon atoms. In some embodiments, cycloaliphatic (or carbocycle or cycloalkyl) refers to a monocyclic C ₃ -C ₆ hydrocarbon that is fully saturated or that contains one or more units of unsaturation but is not aromatic, having one attachment site to the rest molecules. Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

- 6 045073

As used herein, the term bicyclic ring or bicyclic ring system refers to any bicyclic ring system, that is, a carbocyclic or heterocyclic ring system, that is saturated or has one or more units of unsaturation and has one or more common atoms between two rings in the ring system. Thus, the term includes any valid ring fused system, such as an orthocondensed or spirocyclic system. As used herein, the term heterobicyclic is a subcategory of the term bicyclic, wherein one or more heteroatoms are present on one or more rings of the bicyclic. Such heteroatoms may be present at ring junctions, may optionally be substituted, and may be selected from nitrogen (including N-oxides), oxygen, sulfur (including oxidized forms such as sulfones and sulfonates), phosphorus (including oxidized forms such as phosphates ), boron, etc. In some embodiments, the bicyclic group contains 7-12 ring members and 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. According to the International Union of Pure and Applied Chemistry (IUPAC) definition, a bridge is an unbranched chain of atoms, or an atom or valence bond, connecting two bridgeheads, where a bridgehead is any skeletal atom of a ring system that is bonded to three or more skeletal atoms (with the exception of hydrogen). In some embodiments, the bridged bicyclic group contains 7-12 ring members and 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Such bridged bicyclic groups are well known in the art and include the groups listed below, where each group is attached to the rest of the molecule at any substituted carbon or nitrogen atom. Unless otherwise indicated, the bridging bicyclic group is optionally substituted with one or more substituents specified for aliphatic groups. Additionally or alternatively, any substituted nitrogen of the bridging bicyclic group is optionally substituted. Examples of bicyclic rings include:

n

Examples of bridged bicyclic rings include:

The term lower alkyl refers to a C1-4 straight or branched alkyl group. Exemplary lower alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl and tert-butyl.

The term lower haloalkyl refers to a C1-4 straight or branched alkyl group that is substituted with one or more halogen atoms. The term heteroatom means one or more of oxygen, sulfur, nitrogen, phosphorus or silicon (including any oxidized form of nitrogen, sulfur, phosphorus or silicon; the quaternized form of any basic nitrogen or a substituted heterocyclic ring nitrogen, e.g. N (as in 3,4-dihydro -2H-pyrrolil), NH (as in pyrrolidinyl) or NR+ (as in N-substituted pyrrolidinyl)).

The term unsaturated as used herein means that the moiety contains one or more units of unsaturation.

As used herein, the term divalent C1-4 (or _C1-6 ) saturated or unsaturated, straight or branched hydrocarbon chain refers to bivalent alkylene, alkenylene and alkynylene chains that are straight or branched as defined herein. The term alkylene refers to a divalent alkyl group. An alkylene chain is a polymethylene group, i.e. -(CH ₂ )n-, where n is a positive integer, preferably 1 to 6, 1 to 4, 1 to 3, 1 to 2 or 2 to 3. The substituted alkylene chain is a polymethylene group, in which one or more methylene hydrogen atoms are replaced by a substituent. Suitable substituents include those described below for the substituted aliphatic group.

The term alkenylene refers to a divalent alkenyl group.

A substituted alkenylene chain is a polymethylene group containing at least one double bond in which one or more hydrogen atoms have been replaced by a substituent. Suitable substituents include those described below for the substituted aliphatic group.

As used herein, the term cyclopropylene refers to the bivalent cyclopropyl

- 7 045073 group with the following structure:

The term halogen means F, Cl, Br or I.

The term aryl, used alone or as part of a larger moiety such as aralkyl, aralkoxy or aryloxyalkyl, refers to monocyclic or bicyclic ring systems containing a total of five to fourteen ring units, at least one ring of which is aromatic. , and each ring in this system contains from 3 to 7 ring elements. The term aryl may be used interchangeably with the term aryl ring. In some embodiments, aryl refers to an aromatic ring system that includes, but is not limited to, phenyl, biphenyl, naphthyl, anthracyl, and the like, which may contain one or more substituents. Also included within the scope of the term aryl as used herein is a group in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthymidyl, phenanthridinyl or tetrahydronaphthyl and the like.

The terms heteroaryl and heteroar-, used alone or as part of a larger moiety such as heteroaralkyl or heteroaralkoxy, refer to groups containing from 5 to 10 ring atoms, preferably 5, 6 or 9 ring atoms; containing 6, 10 or 14 π electrons distributed in a cyclic structure; and containing, in addition to carbon atoms, from one to five heteroatoms. The term heteroatom refers to nitrogen, oxygen or sulfur and includes any oxidized form of nitrogen or sulfur and any quaternized form of basic nitrogen. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl and pteridinyl . The terms heteroaryl and heteroar- as used herein also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic or heterocyclyl rings, the radical or attachment site being on the heteroaromatic ring. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazine yl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl and pyrido [2,3-b]-1,4-oxazin-3(4H)-one. The heteroaryl group may be mono- or bicyclic. The term heteroaryl may be used interchangeably with the terms heteroaryl ring, heteroaryl group, or heteroaromatic, all of which include rings that are optionally substituted. The term heteroaralkyl refers to an alkyl group substituted with heteroaryl, wherein the alkyl and heteroaryl moieties are independently optionally substituted. As used herein, the terms heterocycle, heterocyclyl, heterocyclic radical and heterocyclic ring are used interchangeably and refer to a stable 5-7 membered monocyclic or 7-10 membered bicyclic heterocyclic moiety that is either saturated or partially unsaturated and contains, in addition to carbon atoms, one or more heteroatoms, preferably from one to four heteroatoms as defined above. When used in reference to the ring atom of a heterocycle, the term nitrogen includes substituted nitrogen. As an example, in a saturated or partially unsaturated ring containing 0-3 heteroatoms selected from oxygen, sulfur, or nitrogen, said nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl ) or +NR (as in N-substituted pyrrolidinyl). The heterocyclic ring may be connected to its side group at any heteroatom or carbon atom that provides a stable structure, and any of the ring atoms may be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanil, diazepinyl, oxazepine yl, thiazepinil, morpholinil and quinuclidinil. The terms heterocycle, heterocyclyl, heterocyclyl ring, heterocyclic group, heterocyclic moiety and heterocyclic radical are used interchangeably herein and also include groups in which the heterocyclyl ring is fused to one or more aryl, heteroaryl or cycloaliphatic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl or tetrahydroquinolinyl. The heterocyclyl group may be mono- or bicyclic. The term heterocyclylalkyl refers to an alkyl group substituted with heterocyclyl, wherein the alkyl and heterocyclyl moieties are independently optionally substituted.

As used herein, the term partially unsaturated refers to a ring moiety that contains at least one double or triple bond. The term partially unsaturated is intended to include rings containing multiple centers of unsaturation, but the term is not intended to include aryl or heteroaryl moieties as defined herein.

As described herein, the compounds of the present invention may contain optionally substituted moieties. In general, the term substituted, whether optionally preceded by a term or not, means that one or more hydrogen atoms of the specified moiety have been replaced by a suitable substituent. Unless otherwise indicated, an optionally substituted group may contain a suitable substituent at each substitutable position of the specified group, and when more than one position in any particular structure can be substituted by more than one substituent selected from a particular group, the specified substituent may be the same or different in every position. The combinations of substituents provided in the present invention are preferably combinations that provide stable or chemically feasible compounds. The term stable as used herein refers to compounds that are substantially unchanged by conditions to enable their production, detection, and in some embodiments, their isolation, purification, and use for one or more of the purposes disclosed herein.

Suitable monovalent substituents on the substituted carbon atom of the optionally substituted group are independently halogen; -(CH2) ₀ _ ₄ R°; -(CH2) ₀ _ ₄ OR°; -0(CH ₂ ) _o-4 R°, -O-(CH ₂ ) _0-4 C(o)OR°; -(CH ₂ ) _o-4 CH(OR°) ₂ ; -(CH ₂ ) ₀ _ ₄ SR°; -(CH ₂ ) ₀ _ ₄ Ph, which can be replaced by R°; -(CH ₂ ) ₀ . ₄ O(CH ₂ ) ₀ .iPh, which can be substituted with R°; -CH=CHPh, which may be substituted with R°; -(CH ₂ ) ₀ . ₄ O(CH ₂ ) _0.1 -pyridyl, which may be substituted with R°; -NO2; -CN; -N ₃ ; -(CH ₂ ) _o-4 N(R°) ₂ ; -(CH2)o-4N(R°)C(0)R°; -N(R°)C(S)R°; -(CH ₂ )o.:\(R4C(())\R< -N(R ^o )C(S)NR ^o 2; -(CH2)o-4N(R°)C(O)OR°; -N(R°)N(R°)C(O)R°; -N(R ^o )N(R ^o )C(O)NR ^o 2; -N(R°)N(R°)c( O)OR°; -(CH ₂ ) _o-4 C(0)R°; -C(S)R°; -(CH2)o-4C(0)OR°; -(CH2)o-4C(0 )SR°; -(CH2)o-4C(O)OSiR°3; -(CH2)o-40C(0)R°; -OC(0)(CH2)o-4SR°; SC(S)SR° ; -(CH2)o-4SC(0)R°; -(CH2)o-4C(0)NR°2; -C(S)NR°2; -C(S)SR°; -SC(S) SR°; -(CH2)o-40C(0)NR°2; -C(O)N(OR°)R°; -C(O)C(O)R°; -C(O)CH2C(O )R ^o ; -C(NOR°)R°; -(CH2)o-4SSR°; -(CH2)o-4S(0)2R°; -(CH2)o-4S(0)20R°; -( CH2)o-40S(0)2R°; -S(O)2NR°2; -S(O)(NR°)R°; -S(O)2N=C(NR°2)2; -(CH2 )o-4S(0)R°; -N(R ^o )S(O)2NR ^o 2; -N(R ^o )S(O)2R ^o ; -N(OR°)R°; -C(NH )NR ^o 2; -P(O)2R°; -P(O)R°2; -OP(O)R°2; -OP(O)(OR°)2; -SiR° ₃ ; -(С _1-4 straight or branched alkylene)ON(R ^o )2 or -(C1-4 straight or branched alkylene)C(O)O-N(R ^o )2, where each R° may be substituted as defined below, and independently represents hydrogen, a C _1-6 aliphatic group, -CH ₂ Ph, -O(CH ₂ ) _0-1 Ph, -CH ₂ -(5-6 membered heteroaryl ring) or 5-6 membered saturated, a partially unsaturated or aryl ring containing 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur or, regardless of the above definition, two independent R° variants together with the intervening atom(s) form a 3-12 membered saturated , a partially unsaturated or aryl mono or bicyclic ring containing 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, which may be substituted as defined below.

Suitable monovalent substituents on R° (or a ring formed by two independent variants of R° together with intervening atoms) are independently halogen, -(CH2)o-2R'; -(halogen R*); -(CH2)o-20H; -(CH2)o-20R*; -(CH2)o-2CK(OR*)2; -((halogen R*); -CN; -N3; -(CH2)o-2C(O)R·; -(CH2)o-2C(0)OH; -(CH ₂ K ₂ C(O)OR ·; -(CH ₂ KJ -(CH2)o-2SH; -(CH2K2NH2; -(CH ₂ ) _o-2 NHR*; -(CH ₂ ) _o-2 NR* ₂ ; -NO2, -SiR* ₃ ; -OSiR* ₃ ; -C(O)SR*; -(C _1-4 straight or branched alkylene)C(O)OR*; or -SSR* where each R* is unsubstituted or, if preceded by a halogen, R * substituted with only one or more halogens and independently selected from a C _1-4 aliphatic group, -CH ₂ Ph, -O(CH ₂ ) _0-1 Ph or a 5-6 membered saturated, partially unsaturated or aryl ring containing 0- 4 heteroatoms independently selected from nitrogen, oxygen or sulfur Suitable divalent substituents on the saturated carbon atom R° include =O and =S.

Suitable divalent substituents on the saturated carbon atom of an optionally substituted group include the following: =O; =S; =NNR*2; =NNHC(O)R*; = NNHC(O)OR*; =NNHS(O) ₂ R*; =NR*; =NOR*; -O(C(R* ₂ )) _2-3 O-; or -S(C(R* ₂ )) _2-3 S-; wherein R* is in each independent instance selected from hydrogen, a C _1-6 aliphatic group which may be substituted as defined below, or an unsubstituted 5-6 membered saturated, partially unsaturated or aryl ring containing 0-4 heteroatoms, independently selected from nitrogen, oxygen or sulfur. Suitable divalent substituents that are bonded to the vicinal substituted carbon atoms of an optionally substituted group include: -O(CR* ₂ ) _2-3 O-, wherein R* is in each independent instance selected from hydrogen, a C _1-6 aliphatic group which may be substituted as defined below, or an unsubstituted 5-6 membered saturated, partially unsaturated or aryl ring containing 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

Suitable substituents on the aliphatic group R* include halogen, -R*; -(halogen R*); -OH, -OR*; -((halogen R*); -CN; -C(O)OH; -C(O)OR*; -NH2; -NHR*; -NR*2; or -NO2; where each R* is unsubstituted or, if preceded by a halogen, R* is substituted with only one or more halogens and

- 9 045073 independently represents a C1- ₄ aliphatic group, -CH2Ph; -O(CH2) _0-1 Ph; or a 5-6 membered saturated, partially unsaturated or aryl ring containing 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

Suitable substituents on the nitrogen atom of the optionally substituted group being replaced include -R ¹ ; -NR^; -C(O)R ^f ; -C(O)OR ^f ; -C(O)C(O)R ^f ; -C(O)CH2C(O^; -8(0^; -SCO^NR^; -C(S)NR ^f 2; -C(NH)NR ¹ 2 or -N(R ¹ )S(O) 2R ¹ ; where each R ¹ independently represents hydrogen, a C1-6 aliphatic group, which may be substituted as defined below, unsubstituted -OPh or an unsubstituted 5-6 membered saturated, partially unsaturated or aryl ring containing 0-4 heteroatoms , independently selected from nitrogen, oxygen or sulfur, or, regardless of the above definition, two independent variants of R ¹ together with the intervening atom(s) form an unsubstituted 3-12 membered saturated, partially unsaturated or aryl mono- or a bicyclic ring containing 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Suitable substituents on the aliphatic group R ¹ are independently halogen, -R*; -(halogen R*); -OH; -OR*; -0 (halogen R*); -CN; -C(O)OH; -C(O)OR*; -NH2; -NHR*; -NR* ₂ or -NO2; where each R* is unsubstituted or, if preceded is halogen, R* is substituted with only one or more halogens, and independently represents a C1- ₄ aliphatic group, -CH ₂ Ph; -O(CH ₂ ) ₀ -1Ph or 5-6-membered saturated; a partially unsaturated or aryl ring containing 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. As used herein, the term pharmaceutically acceptable salt refers to those salts which, in the judgment of those skilled in the art of medicine, are suitable for use in contact with tissues of humans and lower animals without undesirable toxicity, irritation, allergic reaction, or the like, and having an adequate benefit/benefit ratio. risk. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, and the contents of said publication are incorporated herein by reference. Pharmaceutically acceptable salts of the compounds of the present invention include salts derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable, non-toxic acid addition salts are amino salts prepared using inorganic acids such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloric acid, or using organic acids such as acetic acid, oxalic acid , maleic acid, tartaric acid, citric acid, succinic acid or malonic acid, or other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, g facerophosphate, gluconate, hemisulfate, heptanoate , hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalene sulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate , propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate salts and the like.

Salts derived from suitable bases include alkali metal, alkaline earth metal, ammonium and N ⁺ ( _C1-4 alkyl) ₄ salts. Typical alkali or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium and the like. Other pharmaceutically acceptable salts, as appropriate, include salts containing non-toxic ammonium, quaternary ammonium cations and amine cations prepared using counterions such as halide, hydroxide, carboxylate, sulfate, phosphate, nitrate, lower alkyl sulfonate and arylsulfonate.

It is also intended that, unless otherwise indicated, the structures presented herein include all isomeric (eg, enantiomeric, diastereomeric, and geometric (or conformational)) forms of the stated structures; for example, R- and S-configurations for each center of asymmetry, Z- and E-isomers according to the position of the substituents relative to the double bond, and conformational Z- and E-isomers. Thus, individual stereochemical isomers, as well as mixtures of enantiomers, diastereomers and geometric (or conformational) isomers of the compounds of the present invention are included within the scope of the present invention. Unless otherwise indicated, all tautomeric forms of the compounds of the present invention are included within the scope of the present invention. In addition, it is also intended that, unless otherwise indicated, the structures presented herein also include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the structures of the present invention involving the replacement of hydrogen by deuterium or tritium or the replacement of carbon by ¹³ C- or ¹⁴ C-rich carbon are within the scope of the present invention. Such connections are suitable for applications such as

- 10 045073 measures, as analytical tools, as probes in biological research or as therapeutic agents according to the present invention. As used herein, the term inhibitor is defined as a compound that binds to and/or inhibits the interaction of TCRNck with measurable affinity. In some embodiments, the inhibitor has an IC50 and/or binding constant of less than about 100 μM, less than about 50 μM, less than about 1 μM, less than about 500 nM, less than about 100 nM, less than about 10 nM, or less than about 1 nM.

The terms measurable affinity and measurably inhibit as used herein mean to measurably alter TCR activity upon interaction when comparing a sample containing a compound of the present invention or a composition thereof, TCR and Nck, and an equivalent sample containing TCR and Nck and not containing the compound or composition thereof.

The term modulate and modulation as used herein means influencing or changing activity. Modulation includes stabilization, destabilization, amplification and suppression.

The term protein-protein interaction (PPI) as used herein means the physical contact of two or more protein molecules with high specificity. It will be apparent to one of ordinary skill in the art that such interactions include short transient interactions as well as stable interactions.

Description of illustrative embodiments.

Some embodiments of the present invention provide a compound of formula I:

or a pharmaceutically acceptable salt thereof, wherein

R ¹ is R, halogen, -CN, -OR or -N(R)2;

R2 is R, halogen, -C(O)N(R)2 or -N(R)2;

R ³ represents hydrogen or an electron-withdrawing group;

each R independently represents hydrogen or an optionally substituted group selected from Ci_ ₆ aliphatic group, 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring, phenyl, 4-8 membered saturated or partially unsaturated monocyclic heterocyclic ring containing 1-2 a heteroatom independently selected from nitrogen, oxygen or sulfur, a 5-6 membered monocyclic heteroaromatic ring containing 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur; or two R groups on the same nitrogen atom are combined with intervening atoms to form an optionally substituted 5-6 membered heterocyclic ring containing 0-1 heteroatom in addition to the nitrogen atom attached to said ring, such heteroatom being oxygen , nitrogen or sulfur;

L ¹ represents a covalent bond or C1_ ₄ bivalent straight or branched saturated or unsaturated hydrocarbon chain in which 1-2 methylene chain units are independently and optionally replaced by -O-, -C(O)-, -C(O)O- , -OS(O)-, -C(S)-, -C(R) ₂ -, -C(F) ₂ -, -N(R)-, -C(O)N(R)-, - RNC(O)-, -OC(O)N(R)-, -N(R)C(O)N(R) - or -Су-;

L ² represents a covalent bond or C1_ ₄ bivalent straight or branched saturated or unsaturated hydrocarbon chain in which 1-2 methylene chain units are independently and optionally replaced by -O-, -C(O)-, -C(O)O- , -OC(O)-, -C(S)-, -C(R) ₂ -, -CH(R)-, -C(F) ₂ -, -N(R)-, -C(O) N(R)-, -RNC(O)-, -OC(O)N(R)- or -N(R)C(O)N(R)-; and each Cy is independently a bivalent optionally substituted 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring, an optionally substituted phenylene, an optionally substituted 4-8 membered saturated or partially unsaturated monocyclic heterocyclic ring containing 1-3 heteroatoms independently selected from nitrogen, oxygen or sulfur, an optionally substituted 5-6 membered monocyclic heteroaromatic ring containing 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

As defined above and described herein, R ¹ is R, halogen, -CN, -OR or -N(R)2.

In some embodiments, R ¹ is hydrogen.

In some embodiments, R ¹ is halogen or -CN.

In some embodiments, R ¹ is R, -OR, or -N(R)2.

- 11 045073

some implementation options R ¹ some implementation options R ¹ some implementation options R ¹ represents -CH ₃ , -CD ₃ , -CH(CH ₃ ) ₂ , represents ^?

or

is R. In some embodiments, R is optionally substituted Ci. ₆ aliphatic group. In some embodiments, R is an optionally substituted Ci-balkyl. In some embodiments, R is C^alkyl, optionally substituted with halogen or -COOH. In some embodiments, R is C1. ₆ alkyl, 1-6 times substituted with halogen. In some embodiments, R is C1. ₆ alkyl, 1 times substituted -COOH.

In some embodiments, R ¹ is

In some embodiments, R ¹ is

In some embodiments, R ¹ is

In some embodiments, R ¹ is phenyl.

In some embodiments, R ¹ is

In some embodiments, R is selected from the options shown in table. 1 below.

As defined above and described herein, R ² is R, halogen, -C(O)N(R) ₂ or -N(R) ₂ .

In some embodiments, R ² is hydrogen. In some embodiments, R ² is R. In some embodiments, R ² is halogen. In some embodiments, R ² is -C(O)N(R) ₂ or -N(R) ₂ .

In some embodiments, R ² is -C(O)N(R)2 or -N(R)2, wherein each R is independently hydrogen or an optionally substituted group selected from Ci.6 aliphatic group, 3-8- membered saturated or partially unsaturated monocyclic carbocyclic ring, phenyl, 4-8 membered saturated or partially unsaturated monocyclic heterocyclic ring containing 1-2 heteroatoms independently selected from nitrogen, oxygen or sulfur, 5-6 membered monocyclic heteroaromatic ring containing 1 -4 heteroatoms independently selected from nitrogen, oxygen or sulfur. In some embodiments, R ² is -C(O)N(R)2 or -N(R) ₂ , wherein the two R groups at the nitrogen atom are combined with intervening atoms to form an optionally substituted 5-6 membered heterocyclic ring containing 0-1 heteroatom in addition to a nitrogen atom attached to said ring, such heteroatom being oxygen, nitrogen or sulfur. In some embodiments ^{, R2} is -N(R) ₂ , wherein said two R groups at the nitrogen atom are combined with intervening atoms to form a 5-6 membered heterocyclic ring containing no heteroatom in addition to the nitrogen atom attached to said ring , and such a 5-6 membered heterocyclic ring is optionally substituted 1-6 times by Ci. ₃ aliphatic group or halogen.

In some embodiments, R ² is -NCH ₂ CH ₃ or -N(CH ₂ ) ₂ N(CH ₃ ) ₂ .

In some embodiments, R ² is

In some embodiments, R ² is

or

In some embodiments, R ² is

In some embodiments, R ² is selected from the options shown in table. 1 below.

As defined above and described herein, R ³ represents hydrogen or an electron-withdrawing group.

In some embodiments, R ³ is hydrogen.

In other embodiments, R ³ is an electron-withdrawing group. Electron-withdrawing groups are well known to those skilled in the art and include those described in detail in March's Advanced Organic Chemistry, ^5th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001.

In some embodiments, R ³ is an electron-withdrawing group selected from halogen, -CN, -NO ₂ or a C _1-4 aliphatic group substituted with 1-9 halogen atoms.

In some embodiments, R ³ is halogen.

In some embodiments, R ³ is fluorine, chlorine, or bromine.

In some embodiments, R ³ is fluorine.

In some embodiments, R ³ is selected from ^or

F^F

In some embodiments, R ³ is

In some embodiments, R ³ is selected from the options shown in table. 1 below.

As defined above and described herein, R independently represents hydrogen or an optionally substituted group selected from C _1-6 aliphatic group, 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring, phenyl, 4-8 membered saturated or partially unsaturated a monocyclic heterocyclic ring containing 1-2 heteroatoms independently selected from nitrogen, oxygen or sulfur, a 5-6 membered monocyclic heteroaromatic ring containing 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur; or two R groups on the same nitrogen atom are combined with intervening atoms to form an optionally substituted 5-6 membered heterocyclic ring containing 0-1 heteroatom in addition to the nitrogen atom attached to said ring, such heteroatom being oxygen , nitrogen or sulfur.

In some embodiments, R is selected from the options shown in table. 1 below.

As defined above and described herein, L ¹ is a covalent bond or C1-4 bivalent straight or branched saturated or unsaturated hydrocarbon chain in which 1-2 methylene chain units are independently and optionally replaced by -O-,

-C(O)-, -C(O)O-, -OC(O)-, -C(S)-, -C(R)2-, -C(F)2-, -N(R) -, -C(O)N(R)-, -RNC(O)-, -OC(O)N(R)-, ¹

-N(R)C(O)N(R)- or -Су-. In some embodiments, L ¹ is a covalent bond. In some embodiments, L ¹ is a C1-4 bivalent straight or branched saturated or unsaturated hydrocarbon chain in which 1-2 methylene units of the chain are independently and optionally replaced with -O-, -C(O)-, -C(O) O-, -OC(O)-, -C(S)-, -C(R)2-, -C(F)2-, -N(R)-, -C(O)N(R)- , -RNC(O)-, -OC(O)N(R)- or -N(R)C(O)N(R)-.

In some embodiments, L ¹ is a C1-4 bivalent straight or branched saturated or unsaturated hydrocarbon chain in which 1-2 methylene units of the chain are independently and optionally replaced by -O-, -C(R) ₂ -, -C(O )N(R)- or -RNC(O)-.

In some embodiments, L ¹ is a C1-4 bivalent straight saturated or unsaturated hydrocarbon chain in which 1-2 methylene chain units are independently and optionally replaced with -O-, -C(R)2-, -C(O)N (R)- or -RNC(O)-.

In some embodiments, L ¹ is a C1-4 bivalent branched saturated or unsaturated hydrocarbon chain in which 1-2 methylene units of the chain are independently and optionally replaced by -O-, -C(R) ₂ -, -C(O)N (R)-or -RNC(O)-. In some embodiments, L ¹ is -O-.

In some embodiments, L ¹ is

In some embodiments, L ¹ is or o

In some embodiments, L ¹ is selected from the options shown in table. 1 below.

As defined above and described herein, L ² is a covalent bond or C1-4 bivalent straight or branched saturated or unsaturated hydrocarbon chain in which 1-2 methylene chain units are independently and optionally replaced by -O-,

-C(O)-, -C(O)O-, -OC(O)-, -C(S)-, -C(R)2-, -CH(R)-, -C(F)2 -, -N(R)-, -C(O)N(R)-, -RNC(O)-, -OC(O)N(R)- or -N(R)C(O)N(R )-.

In some embodiments, L ² is a covalent bond.

- 13 045073

In other embodiments, L ² is a C _1-4 bivalent straight or branched saturated or unsaturated hydrocarbon chain in which 1-2 methylene units of the chain are independently and optionally replaced by -O-, -C(O)-, -C(O )O-, -OS(O)-, -C(S)-, -C(R) ₂ -, -CH(R)-,

-C(F)2-, -N(R)-, -C(O)N(R)-, -RNC(O)-, -OC(O)N(R)- or -N(R)C (O)N(R)-.

In some embodiments, L ² is a C _1-4 bivalent straight or branched saturated or unsaturated hydrocarbon chain, where 1 methylene unit of the chain is replaced by -O-, -C(O)-, -C(O)O-, -OC (O)-, -C(R)2-, -C(F)2-, -N(R)-, -C(O)N(R)-, -RNC(O)-.

In some embodiments, L ² is a C _1-4 bivalent straight or branched saturated or unsaturated hydrocarbon chain.

In some embodiments, L ² is -C(O)-.

In some embodiments, L ² is -CH ₂ - or -(CH ₂ ) ₂ -.

In some embodiments, L ² is selected from the options shown in table. 1 below.

As generally defined above, each -Cy- is independently a bivalent optionally substituted 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring, an optionally substituted phenylene, an optionally substituted 4-8 membered saturated or partially unsaturated monocyclic heterocyclic ring containing 1 -3 heteroatoms independently selected from nitrogen, oxygen or sulfur, optionally substituted 5-6 membered monocyclic heteroaromatic ring containing 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, optionally substituted 8-10 membered bicyclic or bridged bicyclic a saturated or partially unsaturated heterocyclic ring containing 1-5 heteroatoms independently selected from nitrogen, oxygen or sulfur or an optionally substituted 8-10 membered bicyclic or bridged bicyclic heteroaromatic ring containing 1-5 heteroatoms independently selected from nitrogen, oxygen or sulfur .

In some embodiments, -Cy- is a bivalent, optionally substituted, 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring. In some embodiments, -Cy- is an optionally substituted phenylene. In some embodiments, -Cy- is an optionally substituted 4-8 membered saturated or partially unsaturated monocyclic heterocyclic ring containing 1-3 heteroatoms independently selected from nitrogen, oxygen or sulfur. In some embodiments, -Su is an optionally substituted 5-6 membered monocyclic heteroaromatic ring containing 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. In some embodiments, -Cy- is an optionally substituted 8-10 membered bicyclic or bridged bicyclic saturated or partially unsaturated heterocyclic ring containing 1-5 heteroatoms independently selected from nitrogen, oxygen or sulfur. In some embodiments, -Cy- is an optionally substituted 8-10 membered bicyclic or bridged bicyclic heteroaromatic ring containing 1-5 heteroatoms independently selected from nitrogen, oxygen or sulfur.

n/M g Y

In some embodiments, -Cy- is or +.

In some embodiments, -Su- is selected from the options shown in table. 1 below.

In some embodiments, the present invention provides a compound of Formula II:

R ³

R ¹

'R ² or a pharmaceutically acceptable salt thereof, wherein each R ¹ , R ² , R ³ , R, L ¹ , L ² and -Cy- are defined above and described in the embodiments herein, either individually or in combination .

In some embodiments, the present invention provides a compound of Formula III:

or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R , L ¹ , L ² , and -Cy- are each as defined above and described in the embodiments herein, either individually or in combination.

In some embodiments, the present invention provides a compound of formula IV- 14 045073 a, IV-b, IV-c or IV-d:

ants implemented herein, either individually or in combination.

In some embodiments, the present invention provides a compound of formula Va, V-b, V-c, or V-d:

or a pharmaceutically acceptable salt thereof, wherein each R ¹ and R is as defined above and described in the embodiments herein, either individually or in combination. In some embodiments, the present invention provides a compound of any of formulas VI-a, VI-b, VI-c and VI-d:

implementations in this document, either individually or in combination. In some embodiments, the present invention provides a compound of Formula VII:

R ³

VII or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ³ , R and L ² are each as defined above and described in the embodiments herein, either individually or in combination.

In some embodiments, the present invention provides a compound of Formula VIII:

- 15 045073

R ³

BUT

R ²

VIII or a pharmaceutically acceptable salt thereof, wherein R ² , R ³ , L ¹ and L ² are each as defined above and described in the embodiments herein, either individually or in combination.

In some embodiments, the present invention provides a compound of Formula IX:

embodiments herein, either individually or in combination.

In some embodiments, the present invention provides a compound of Formula X:

R ³

BUT

R ²

X or a pharmaceutically acceptable salt thereof, wherein R, R ² , R ³ and L ¹ are each as defined above and described in the embodiments herein, either individually or in combination.

In some embodiments, the present invention provides a compound of formula XIa or XI-b:

or a pharmaceutically acceptable salt thereof, wherein R, R ² , R ³ and L ¹ are each as defined above and described in the embodiments herein, either individually or in combination.

In some embodiments, the present invention provides a compound of formula XII:

or a pharmaceutically acceptable salt thereof, wherein R ² , R ³ , L ¹ and L ² are each as defined above and described in the embodiments herein, either individually or in combination.

In some embodiments, the present invention provides a compound of formula XIII:

R ³

R

'R ²

XIII or a pharmaceutically acceptable salt thereof, wherein R ¹ , R ² , R ³ and L ² are each as defined above and described in

- 16 045073 embodiments herein, either individually or in combination.

Examples of compounds according to the present invention are given in table. 1 below.

Table 1

Connection examples

1-13 1-14

- 17 045073

- 18 045073

- 19 045073

- 20 045073

1-82

In some embodiments, the present invention provides the compound listed in table. 1 above, or a pharmaceutically acceptable salt thereof.

Application, composition and administration.

Pharmaceutically acceptable compositions.

According to another embodiment, the present invention provides a composition comprising a compound of the present invention or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant or carrier. The amount of compound in the composition of the present invention is such that it is effective to measurably modulate the interaction between TCR and Nck in a biological sample or patient. In some embodiments, the amount of a compound in a composition of the present invention is such that it is effective to measurably modulate the interaction between TCR and Nck in a biological sample or patient. In some embodiments, a composition of the present invention is formulated for administration to a patient in need of such a composition. In some embodiments, the composition of the present invention is formulated for oral administration to a patient.

The term patient as used herein means an animal, preferably a mammal, and most preferably a human.

- 21 045073

The term pharmaceutically acceptable carrier, adjuvant or vehicle refers to a non-toxic carrier, adjuvant or vehicle that does not interfere with the pharmacological activity of the compound being formulated with it. Pharmaceutically acceptable carriers, adjuvants or carriers that may be used in the composition of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffers such as phosphates , glycine, sorbic acid, potassium sorbate, partial glycerides of a mixture of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, sodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinylpyrrolidone, substances based on cellulose, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene block polymers, polyethylene glycol and lanolin (wool fat).

The compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or by an implantable reservoir. The term parenteral as used herein includes subcutaneous, intravenous, intramuscular, intraarticular, intrasynovial, intrasternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion. Preferably the compositions are administered orally, intraperitoneally or intravenously. Sterile injectable forms of the compositions of the present invention may be an aqueous or oily suspension. These suspensions can be prepared in accordance with methods known in the art, using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic diluent or solvent suitable for parenteral administration, for example a solution in 1,3-butanediol. Acceptable carriers and solvents that may be used include water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile fixed oils are usually used as a solvent or suspending medium.

For this purpose, any light (bland) fixed oil can be used, including synthetic mono or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives can be used to prepare injectable forms, examples being natural pharmaceutically acceptable oils such as olive oil or castor oil, in particular in their polyoxyethylated forms. These oily solutions or suspensions may also contain a long-chain alcohol solvent or dispersant, such as carboxymethylcellulose or similar dispersants, which are commonly used in pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants, such as the Tween, Span, and other emulsifying agents or bioavailability enhancers that are commonly used to prepare pharmaceutically acceptable solid, liquid, or other dosage forms, may also be used to formulate the formulations.

The pharmaceutically acceptable compositions of the present invention can be administered orally in any orally acceptable dosage form, including, but not limited to, capsules, tablets, aqueous suspensions or solutions. For oral tablets, carriers typically include lactose and corn starch. Lubricants such as magnesium stearate are also commonly added. For oral administration in capsule form, suitable diluents include lactose and dried corn starch. When aqueous suspensions for oral administration are required, the active ingredient is combined with emulsifying and suspending agents. Certain sweetening, flavoring or coloring agents may also be added if desired.

Alternatively, the pharmaceutically acceptable compositions of the present invention may be administered in the form of suppositories for rectal administration. They can be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature and liquid at rectal temperature and will thus melt in the rectum, releasing the drug. Such materials include cocoa butter, beeswax and polyethylene glycols.

Pharmaceutically acceptable compositions of the present invention can also be administered topically, particularly when the target for treatment includes areas or organs readily accessible for topical administration, including diseases of the eye, skin or lower gastrointestinal tract. Suitable topical formulations can be readily prepared for each of these areas or organs.

Topical administration to the lower gastrointestinal tract can be achieved using a rectal suppository formulation (see above) or a suitable enema formulation. Topical transdermal patches can also be used.

For topical use, the proposed pharmaceutically acceptable compositions may be

- 22 045073 are prepared in the form of a suitable ointment containing the active ingredient suspended or dissolved in one or more vehicles. Carriers for topical application of the compounds of the present invention include, but are not limited to, mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water. Alternatively, the pharmaceutically acceptable compositions provided may be formulated as a suitable lotion or cream containing the active ingredients suspended or dissolved in one or more pharmaceutically acceptable carriers. Suitable carriers include, but are not limited to, mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutically acceptable compositions of the invention may be formulated as micronized suspensions in isotonic, sterilized pH-adjusted saline or as solutions in isotonic, sterilized pH-adjusted saline, with or without the addition of a preservative such as benzalkonium chloride. Alternatively, for ophthalmic use, pharmaceutically acceptable compositions may be formulated as an ointment such as petroleum jelly.

Pharmaceutically acceptable compositions of the present invention can also be administered by nasal aerosol or by inhalation. Such compositions are prepared according to techniques well known in the pharmaceutical formulation art and can be prepared as saline solutions using benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorocarbons and/or other conventional solubilizing or dispersing agents.

Most preferably, the pharmaceutically acceptable compositions of the present invention are formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, the pharmaceutically acceptable compositions of the present invention are administered without food. In other embodiments, the pharmaceutically acceptable compositions of the present invention are administered with food. The amount of a compound of the present invention that can be combined with carrier substances to formulate a single dosage form will vary depending on the host being treated and the particular route of administration. Preferably, the proposed compositions should be prepared in such a way that a dose of 0.01-100 mg/kg body weight/day of inhibitor can be administered to a patient receiving said compositions. It should also be understood that the specific dosage and treatment regimen for any particular patient will depend on many different factors, including the potency of the particular compound used, age, body weight, general health, gender, diet, time of administration, rate of elimination, drug combination, opinion the attending physician and the severity of the specific disease being treated. The amount of a compound of the present invention in a composition will also depend on the particular compound in the composition.

Uses of Compounds and Pharmaceutically Acceptable Compositions The compounds and compositions described herein are generally useful for modulating TCR signaling and T cell activation by modulating the interaction between TCR and Nck.

The activity of the compound used according to the present invention as a modulator of TCR-Nck interaction can be studied in vitro, in vivo or in a cell line. In vitro assays include, for example, assays for determining T-lymphocyte proliferation (eg, Tse, W.T. et al., Transplantation, 2003, 75(3): 389-97, the contents of which are incorporated herein by reference); determining actin cytoskeleton polymerization induced in T cells following TCR stimulation (eg, Fuller, C.L. et al., Immunol. Rev. 2003, 292: 220-36, the contents of which are incorporated herein by reference); and determining TCR stimulation-induced cytokine secretion by T cells (eg, Finco, D. et al., Cytokine, 2014, 66(2): 143-55, the contents of which are incorporated herein by reference). In vivo assays include standard animal models of immune and autoimmune diseases that are well known and state of the art, such as, for example, delayed-type hypersensitivity models (e.g., Kudlacz, E. et al., Am. J. Transplant., 2004, 4(1): 51-7, the contents of the specified source are incorporated into this description by reference); rheumatoid arthritis models (eg, Holmdahl, R. et al., APMIS, 1989, 97(7): 575-84, the contents of which are incorporated herein by reference); models of multiple sclerosis (experimental autoimmune encephalomyelitis) (eg, Gonzalez-Rey, E. et al., Am. J. Pathol. 2006, 168(4): 1179-88, the contents of which are incorporated herein by reference); and models of graft rejection (see, for example, the various animal models described in the above references, incorporated herein by reference, in relation to the treatment of graft rejection). As used herein, the terms treatment, treat and treating refer to achieving regression, alleviation, delay in onset or suppression of the progression of a disease or disorder or one or more of its symptoms described herein. In some countries

- 23 045073 ant implementation treatment can be prescribed after the development of one or more symptoms. In other embodiments, treatment may be administered in the absence of symptoms. For example, treatment may be administered to a predisposed individual prior to the onset of symptoms (eg, based on a history of symptoms and/or genetic or other predisposition factors). Treatment may also be continued after symptoms have disappeared, for example to prevent or delay their recurrence.

The present compounds are modulators of the TCR-Nck interaction and are accordingly suitable for the treatment of one or more disorders associated with TCR activity. Thus, in some embodiments, the present invention provides a method of treating a TCR-Nck mediated disorder, comprising the step of administering to a patient in need thereof a compound of the present invention or a pharmaceutically acceptable composition thereof. As used herein, the terms TCR-Nck-mediated disorders, diseases and/or conditions mean any disease or other deleterious condition in which the TCR has been shown to play a role. Accordingly, another embodiment of the present invention relates to the treatment or reduction of the severity of one or more diseases in which the TCR has been identified as having a role. Such TCR-Nck-mediated disorders include, but are not limited to, autoimmune and inflammatory disorders; transplant related disorders; proliferative disorders and neurological disorders. (See, for example, O'Sea, J. et al., Nat. Rev. Drug Doscpv. 2004, 3(7): 555-64; Cetkovic-Cvrlje, M. et al., Curr. Pharm. Des. 2004, 10(15): 1767-84; Cetkovic-Cvrlje, M. et al., Arch. Immunol. Ther. Exp. 2004, 52(2): 69-82).

Autoimmune and inflammatory disorders.

As used herein, the term autoimmune and inflammatory disorder refers to those diseases, pathologies or conditions that occur when the body's systems are attacked by that body's own immune system. Targets of autoimmune interactions can range from the cellular level (eg, myelin basic protein in multiple sclerosis or thyroid-stimulating hormone receptor in Graves' disease) to organ-specific effects in rheumatoid arthritis or Crohn's disease to systemic effects seen in systemic lupus erythematosus. Some of the phenomena that have been recognized as causes of autoimmune diseases include overexpression of cytokines such as TNF-α, IL2 or IL-2 receptor in inflammatory bowel disease, or underexpression (IL-10 underexpression in type 1 diabetes), expression defects alleles (human leukocyte antigen class I locus B alleles 27 (HLA Class I B27) in ankylosing spondylitis), altered expression of proteins involved in the regulation of apoptosis (underexpression of Fas in autoimmune lymphoproliferative syndrome type I (ALPS 1). For more information about autoimmune diseases, see Harrison's Principles of Internal Medicine, 16th ed., McGraw-Hill, N.Y., 2005, chapter 295.

In some embodiments, the autoimmune or inflammatory disorder is Addison's disease, agammaglobulinemia, alopecia areata, alopecia universalis, amyloidosis, ankylosing spondylitis, anti-glomerular basement membrane antibody/anti-tubular basement membrane antibody (anti-GBM/anti-TBM) nephritis. , antiphospholipid syndrome, autoimmune angioedema, autoimmune vegetative dystonia, autoimmune encephalomyelitis, autoimmune hepatitis, autoimmune inner ear disease (Meniere's disease), autoimmune myocarditis, autoimmune pancreatitis, autoimmune retinopathy, autoimmune urticaria, axonal and neuronal neuropathy, Balo's disease, Behçet's disease benign mucosal pemphigoid, bullous pemphigoid, Castleman disease, celiac disease, Chagas disease, chronic inflammatory demyelinating polyneuropathy, chronic recurrent multifocal osteomyelitis, Churg-Strauss syndrome, cicatricial pemphigoid, Cogan syndrome, cold agglutinin disease, congenital heart block, Coxsackie myocarditis, CREST- syndrome (calcinosis, Raynaud's disease, esophageal dysmotility, sclerodactyly and telangiectasia), Crohn's disease, dermatitis herpetiformis, dermatomyositis, Devic's disease (neuromyelitis of the optic nerve), discoid lupus, Dressler's syndrome, endometriosis, eosinophilic esophagitis, eosinophilic fasciitis, erythema nodosum, essential mixed cryoglobulinemia, Evans syndrome, fibromyalgia, fibrosing alveolitis, giant cell arteritis, giant cell myocarditis, glomerulonephritis, Goodpasture's syndrome, granulomatosis with polyangiitis, Graves' disease, Guillain-Barré syndrome, Hashimoto's thyroiditis, hemolytic anemia, Henoch-Schönlein purpura, gestational herpes or gestational pemphigoid , hypogammaglobulinemia, IgA nephropathy, IgG4-related sclerosing disease, immune thrombocytopenic purpura, inclusion body myositis, inflammatory bowel disease, interstitial cystitis, juvenile arthritis, juvenile diabetes, juvenile myositis, Kawasaki disease, Lambert-Eaton syndrome, leukocytoclastic vasculitis, red lichen planus, lichen sclerosus, woody conjunctivitis, IgA-dependent linear dermatosis, lupus erythematosus, chronic Lyme disease, Lyme neuroborreliosis, microscopic polyangiitis, mixed connective tissue disease, Moray ulcer, Mucha-Habermann disease,

- 24 045073 multiple sclerosis, myasthenia gravis, myositis, neuromyelitis of the optic nerve, neutropenia, ocular cicatricial pemphigoid, optic neuritis, palindromic rheumatism, PANDAS syndrome (childhood autoimmune neuropsychiatric disorders associated with streptococcal infections), paraneoplastic cerebellar degeneration, paroxysmal nocturnal hemoglobinuria, Parry-Romberg syndrome, parsplanitis, Personage-Turner syndrome, pemphigus, peripheral neuropathy, perivenous encephalomyelitis, pernicious anemia, POEMS syndrome (polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, skin lesions), polyarteritis nodosa, polyglandular syndromes I, II and III types, polymyalgia rheumatica, polymyositis, post-infarction syndrome, post-pericardiotomy syndrome, primary biliary cirrhosis, primary sclerosing cholangitis, progesterone dermatitis, psoriasis, psoriatic arthritis, true erythrocyte aplasia, pyoderma gangrenosum, Raynaud's phenomenon, reactive arthritis, reflex sympathetic dystrophy , relapsing polychondritis , retroperitoneal fibrosis, rheumatic fever, rheumatoid arthritis, sarcoidosis, Schmidt's syndrome, scleritis, scleroderma, Sjögren's syndrome, autoimmune response against sperm and testicular antigens (sperm and testicular autoimmunity), stiff person syndrome, subacute bacterial endocarditis (SBE) ), Susak syndrome, sympathetic ophthalmia, Takayasu arteritis, temporal arteritis, thrombocytopenic purpura (TTP), Tolosa-Hunt syndrome, transverse myelitis, type 1 diabetes, ulcerative colitis, undifferentiated connective tissue disease, uveitis, vasculitis, vitiligo and Wegener's granulomatosis (or granulomatosis with polyangiitis).

Disorders associated with transplantation.

In some embodiments, the disorder is related to the transplantation. In some embodiments, the transplant disorder is graft rejection. In some embodiments, the transplant disorder is graft-versus-host disease.

Proliferative disorders.

In some embodiments, the disorder is a proliferative disorder. In some embodiments, the proliferative disorder is cancer.

As used herein, the term cancer refers to cells that have the ability to grow autonomously, that is, an abnormal status or condition characterized by rapid cell proliferation. The term includes all types of cancerous tumors or oncogenic processes, tissues with metastases or cells, tissues or organs that have undergone malignant transformation, regardless of histopathological type or stage of invasiveness. The term tumor as used herein refers to cancer cells, eg, a formation from cancer cells. The term carcinoma is known in the art and refers to malignancies of epithelial or endocrine tissues, including respiratory system carcinomas, gastrointestinal carcinomas, genitourinary carcinomas, testicular carcinomas, breast carcinomas, prostate carcinomas, endocrine system carcinomas and melanomas. Adenocarcinoma refers to carcinoma that develops from glandular tissue, or carcinoma in which the tumor cells form recognizable glandular structures. The term sarcoma is known in the art and refers to malignant tumors of mesenchymal origin.

Cancers include, but are not limited to, leukemias (eg, acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, acute myeloid leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroid leukemia, chronic leukemia, chronic myelocytic leukemia h, chronic lymphocytic leukemia), polycythemia vera, lymphoma (eg, Hodgkin's disease or non-Hodgkin's lymphoma), Waldenström's macroglobulinemia, multiple myeloma, heavy chain disease, and solid tumors such as sarcomas and carcinomas (eg, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer, ovarian cancer, prostate cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma , sweat cell carcinoma glands, sebaceous carcinoma, papillary carcinoma, papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms tumor, cervical cancer, uterine cancer, ovarian cancer chka , lung carcinoma, small cell lung carcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma, meningioma, melanoma, neuroblastoma and retinoblastoma).

In some embodiments, the proliferative disorder is a hematologic cancer. In some embodiments, the proliferative disorder is leukemia. In some embodiments, the leukemia is a T-cell leukemia.

Neurological disorders.

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In some embodiments, the disorder is a neurological disorder. In some embodiments, the neurological disorder is Balo's disease, chronic inflammatory demyelinating polyneuropathy, Devic's optic neuromyelitis, acute Marburg multiple sclerosis, multiple sclerosis, Schilder's disease, or perivenous encephalomyelitis.

Suitable compositions.

The pharmaceutically acceptable compositions of the present invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (e.g., by powders, ointments, or drops), buccally, as an oral or nasal spray, or the like. depending on the severity of the infection being treated. In some embodiments, the compounds of the present invention may be administered orally or parenterally in doses ranging from about 0.01 mg/kg to about 50 mg/kg, and preferably from about 1 mg/kg to about 25 mg/kg of the subject's body weight in day, one or more times a day, to obtain the desired therapeutic effect. Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compounds, liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed, peanut, corn, germ, olive, castor and sesame oils), glycerin, tetrahydrofurfuryl alcohol, polyethylene glycols and sorbitan fatty acid esters and mixtures thereof. In addition to inert diluents, oral compositions may also contain adjuvants such as wetting agents, emulsifying and suspending agents, sweeteners, flavoring agents and flavoring agents.

Injectable preparations, for example sterile injectable aqueous or oleaginous suspensions, can be prepared in accordance with the prior art using suitable dispersing or wetting agents and suspending agents. The sterile injectable preparation may also be a sterile injectable solution, suspension or emulsion in a non-toxic parenterally acceptable diluent or solvent, for example a solution in 1,3-butanediol. Acceptable media and solvents that may be used include water, Ringer's solution as defined in the United States Pharmacopoeia (U.S.P.), and isotonic sodium chloride solution. In addition, sterile non-volatile oils are usually used as a solvent or suspending medium. For this purpose, any soft, fixed oil can be used, including synthetic mono- or diglycerides. In addition, fatty acids such as oleic acid are used to prepare injectable drugs.

Injectable formulations can be sterilized, for example, by filtration through a bacteria-retaining filter or by administering sterilizing agents in the form of sterile solid compositions that can be dissolved or dispersed in sterile water or other sterile injection medium prior to use. To prolong the effect of the compound of the present invention, it is often desirable to slow down the absorption of the compound by subcutaneous or intramuscular injection. This can be achieved by using a liquid suspension of crystalline or amorphous material with low water solubility. The rate of absorption of a compound is thus dependent on its rate of dissolution, which in turn may depend on crystal size and crystalline form. Alternatively, delayed absorption of the parenterally administered form of the compound is achieved by dissolving or suspending the compound in an oily medium. Injectable depot forms are prepared by forming microcapsule matrices of the compound in biodegradable polymers such as polylactide-polyglycolide. Depending on the ratio of the amounts of compound to polymer and the nature of the particular polymer used, the rate of release of the compound can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Injectable depot formulations are also prepared by enclosing the compound in liposomes or microemulsions that are compatible with body tissues.

Compositions for rectal or vaginal administration are preferably suppositories, which can be prepared by mixing the compounds of the present invention with suitable non-irritating excipients or carriers, such as cocoa butter, polyethylene glycol or suppository wax that is solid at ambient temperature but liquid at body temperature and therefore soften in the rectum or vaginal cavity, releasing the active compound.

Solid dosage forms for oral administration include capsules, tablets, pills, powders and granules. In such solid dosage forms, the active compound is mixed with at least one inert pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate, and/or a) excipients or bulking agents, such as starches, lactose, sucrose, glucose, mannitol and silicic acid, b) binders such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose and gum, c) humectants such as glycerin, d) disintegrants such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, some silicates and sodium carbonate, f) dissolution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may also contain buffering agents.

Solid compositions of a similar type can also be used as fillers in soft and hard gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like. Solid dosage forms in the form of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings and other coatings well known in the field of pharmaceutical formulations. They may optionally contain opacifying agents and may also be formulated such that they release the active ingredient(s) exclusively or preferentially in a specific part of the intestinal tract, optionally in a delayed manner. Examples of inclusion compositions that can be used include polymeric substances and waxes. Solid compositions of this type can also be used as fillers in soft and hard gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like. The active compounds may also be in the form of microcapsules with one or more of the excipients mentioned above. Solid dosage forms in the form of tablets, dragees, capsules, pills and granules can be prepared with coatings and shells, such as enteric coatings, controlled release coatings, and other coatings well known in the field of pharmaceutical formulations. In such solid dosage forms, the active compound may be mixed with at least one inert diluent such as sucrose, lactose or starch. Such dosage forms may also contain, as is commonly practiced, additional substances other than inert diluents, such as tabletting lubricants and other tabletting auxiliaries such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets and pills, dosage forms may also contain buffering agents. They may optionally contain opacifying agents and may also be formulated such that they release the active ingredient(s) exclusively or preferentially in a specific part of the intestinal tract, optionally in a delayed manner. Examples of inclusion compositions that can be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants or patches. The active ingredient is mixed under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservatives or buffers that may be required. A composition for ophthalmic use, eye drops and ear drops are also included within the scope of the present invention. In addition, the present invention provides for the use of transdermal patches, which have the additional advantage of providing controlled delivery of the compound to the body. Such dosage forms can be prepared by dissolving or dispersing the compound in a suitable medium. Absorption enhancers can also be used to increase the flow of the compound through the skin. The rate can be controlled either by providing a rate-regulating membrane or by dispersing the compound in a polymer matrix or gel.

The term biological sample as used herein includes, without limitation, cell cultures or extracts thereof; material obtained by biopsy from a mammal, or extracts thereof; and blood, saliva, ooze, feces, seminal fluid, tears or other body fluids or extracts thereof.

Methods of treatment.

In other embodiments, the present invention provides a method of treating a TCR-Nck interaction disorder in a patient in need thereof, comprising the step of administering to said patient a compound of the present invention or a pharmaceutically acceptable composition thereof. In some embodiments, a method of modulating TCR-Nck is used to treat autoimmune and inflammatory disorders; disorders associated with transplantation; proliferative disorders and neurological disorders. In some embodiments, the method of modulating TCR-Nck is used to treat alopecia areata (see, e.g., Petukhova, L. et al., Nature, 2010, 466(7302): 113-17). Accordingly, some embodiments of the present invention provide a method of treating alopecia areata in a patient in need thereof, comprising the step of administering to said patient a compound of the invention.

- 27 045073 or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat ankylosing spondylitis (see, e.g., Smith, J.A., Curr Allergy Asthma Rep. 2015, 15(1): 489). Accordingly, some embodiments of the present invention provide a method of treating ankylosing spondylitis in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat asthma (see, e.g., Robinson, D.S., J. Allergy Clin. Immunol., 2010, 126(6): 1081-91). Accordingly, some embodiments of the present invention provide a method of treating asthma in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat autoimmune hepatitis (see, for example, Manns, M.P. et al., Hepatology, 2010, 51(6), 2193-213. Accordingly, in some embodiments of the present invention, a method is provided treating autoimmune hepatitis in a patient in need thereof, comprising the step of administering to said patient the proposed compound or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat autoimmune lymphoproliferative syndrome (ALPS) (see, for example, Sneller, M.C. et al., Curr. Opin. Rheumatology, 2003, 15(4) 417-21). Accordingly, some embodiments of the present invention provide a method of treating ALPS in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof. In some embodiments, the method of modulating TCR-Nck is used in autoimmune myocarditis (see, for example, Caforio, A.L. and Iliceto, S., Curr. Opin. Cardiol., 2008, 23(3): 219-26). Accordingly, some embodiments of the present invention provide a method of treating autoimmune myocarditis in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof. In some embodiments, the method of modulating TCR-Nck is used for autoimmune orchitis (see, e.g., Silva, C.A. et al., Autoimmun Rev., 2014, 13(4-5): 431-34). Accordingly, some embodiments of the present invention provide a method of treating autoimmune orchitis in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, the method of modulating TCR-Nck is used in autoimmune pancreatitis (see, e.g., Fan, B. G. and Andren-Sandberg, A., N. Am. J. Med. Sci. 2009, 1(2): 148-51 ). Accordingly, some embodiments of the present invention provide a method of treating autoimmune pancreatitis in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat atopic dermatitis (see, e.g., Nograles, K.E. et al., J. Allergy Clin. Immunol., 2009, 123(6): 1244-52). Accordingly, some embodiments of the present invention provide a method of treating systemic atopic dermatitis in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof. In some embodiments, a method of modulating TCR-Nck is used to treat Behçet's disease (see, e.g., Direskeneli, H., Genetics Research International, 2013, Article ID 249157 doi:10.1155/2013/249157). Accordingly, some embodiments of the present invention provide a method of treating Behçet's disease in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof. In some embodiments, a method of modulating TCR-Nck is used to treat Castleman's disease (see, for example, Al-Maghrabi, J. et al., Histopathology, 2005, 48(3): 233-38). Accordingly, some embodiments of the present invention provide a method of treating Castleman's disease in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat celiac disease (see, e.g., Mazzarella, G., World J. Gastroenterol., 2015, 21(24): 7349-56). Accordingly, some embodiments of the present invention provide a method of treating celiac disease in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat chronic inflammatory demyelinating polyneuropathy (see, e.g., Notturno, F. et al., J. Neuroimmunol. 2008, 197(2): 124-7). Accordingly, some embodiments of the present invention provide a method of treating chronic inflammatory demyelinating polyneuropathy in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

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In some embodiments, a method of modulating TCR-Nck is used to treat Cogan's syndrome (see, e.g., Greco, A. et al., Autoimmunity Rev. 2013, 12(3): 396-400). Accordingly, some embodiments of the present invention provide a method of treating Cogan's syndrome in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat Churg-Strauss syndrome (see, e.g., Guida, G. et al., Clin. Immunol., 2008 128(1): 94-102). Accordingly, some embodiments of the present invention provide a method of treating Churg-Strauss syndrome in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat Crohn's disease (see, e.g., Roche, J.K. et al., J. Clin. Invest. 1985, 75(2):522-530; Marks, D. J. and Segal, A. W. J. Pathol. 2008, 214(2): 260-66; Cobrin, G. M. and Abreu, M. T. Immunol. Rev. 2005, 206(1): 277-95). Accordingly, some embodiments of the present invention provide a method of treating Crohn's disease in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat Evans syndrome (see, for example, Teachery, D.T. et al., Blood, 2004, 105(6):2443-48). Accordingly, some embodiments of the present invention provide a method of treating Evans syndrome in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat inclusion body myositis (see, e.g., Kitazawa, M. et al., J. Neuroscience, 2009, 29(19): 6132-41). Accordingly, some embodiments of the present invention provide a method of treating inclusion body myositis in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof. In some embodiments, a method of modulating TCR-Nck is used to treat inflammatory bowel disease (see, e.g., Zenewicz, L.A. et al., Trends Mol. Med., 2009, 15(5): 199-207). Accordingly, some embodiments of the present invention provide a method of treating inflammatory bowel disease in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat Kawasaki disease (see, for example, Onouchi, Y. et al., Nature Genetics, 2008, 40: 35-42). Accordingly, some embodiments of the present invention provide a method of treating Kawasaki disease in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat Lyme disease (chronic) (see, e.g., Singh, S. K. and Girschick, H. J. Paediatric Rheumatology, 2004, 10(7): 598-614; Raveche, E. S. et al. , J Clin Microbiol 2005, 43(2): 850-56). Accordingly, some embodiments of the present invention provide a method of treating Lyme disease (chronic) in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat multiple sclerosis (see, e.g., Babbe, H. et al., J. Exp. Med., 2000, 192(3): 393-404; Dai, K. Z. et al., Genes Immun. 2001, 2(5): 263-8). Accordingly, some embodiments of the present invention provide a method of treating multiple sclerosis in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat myasthenia gravis (see, e.g., Meriggioli, M.N. and Sanders, D.B.S., Lancet Neurology, 2009, 8(5): 475-90). Accordingly, some embodiments of the present invention provide a method of treating myasthenia gravis in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat psoriasis (see, e.g., Cai, Y. et al., Cell Mol. Immunol., 2012, 9(4): 302-09).

Accordingly, some embodiments of the present invention provide a method of treating psoriasis in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat psoriatic arthritis (see, e.g., Choy, E., Curr. Rheumatol. Rep. Exp., 2007, 9(6): 437-41). Accordingly, some embodiments of the present invention provide a method of treating psoriatic arthritis in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

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In some embodiments, a method of modulating TCR-Nck is used to treat rheumatoid arthritis (see, e.g., Soret, A.R. et al., Clin. Exp. Rheumatol., 2007, 25(5): S4-11). Accordingly, some embodiments of the present invention provide a method of treating rheumatoid arthritis in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat systemic lupus erythematosus (see, e.g., Crispin, J.C. et al., J. Immunol., 2008, 181(12): 8761-66; Linterman, M.A. et al. , J Exp Med 2009, 206(3): 561-76). Accordingly, some embodiments of the present invention provide a method of treating systemic lupus erythematosus in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat type I diabetes (see, for example, Roep, VO, Diabetologia, 46(3): 305-21). Accordingly, some embodiments of the present invention provide a method of treating type I diabetes in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat ulcerative colitis (see, for example, Kappeler, A. and Mueller, C, Histol Histopathol., 2000, 15(1): 167-72). Accordingly, some embodiments of the present invention provide a method of treating ulcerative colitis in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat uveitis (see, e.g., Horai, R. et al., Immunity, 2015, 43(2): 343-53). Accordingly, some embodiments of the present invention provide a method of treating uveitis in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat vitiligo (see, for example, Van den Wijngaard, R. et al., Lab Invest. 2000, 80(8): 1299-309). Accordingly, some embodiments of the present invention provide a method of treating vitiligo in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat transplant rejection (see, e.g., Issa, F. et al., Expert Rev. Clin. Immunol. 2010, 6(1): 155-69). Accordingly, some embodiments of the present invention provide a method of treating transplant rejection in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof. In some embodiments, a method of modulating TCR-Nck is used to treat granulomatosis with polyangiitis (Wegener's granulomatosis) (see, e.g., Morgan, M.D. et al., Arthritis & Rheumatism, 2011, 63(7): 2127-37). Accordingly, some embodiments of the present invention provide a method of treating granulomatosis with polyangiitis (Wegener's granulomatosis) in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

In some embodiments, a method of modulating TCR-Nck is used to treat hematologic cancer. Accordingly, some embodiments of the present invention provide a method of treating hematologic cancer in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof. In some embodiments, a method of modulating TCR-Nck is used to treat graft rejection (see, e.g., Issa, F. et al., Expert Rev. Clin. Immunol. 2010, 6(1): 155-69). In some embodiments, a method of modulating TCR-Nck is used to treat graft-versus-host disease (see, e.g., W.D., Nature Rev. Immunology, 2007, 7: 340-52). Accordingly, some embodiments of the present invention provide a method of treating transplant-related disorder in a patient in need thereof, comprising the step of administering to said patient a compound of the invention or a pharmaceutically acceptable salt thereof.

Combinations.

A compound of the present invention may be administered alone or in combination with one or more other therapeutic compounds, and possible combination therapy may take the form of fixed combinations, or administration of a compound of the present invention and one or more other therapeutic compounds in parallel (staggered), or independently from each other, or combined administration of fixed combinations and one or more other therapeutic compounds. As used herein, additional therapeutic agents that are typically administered to treat a particular disease or condition are known to be suitable for the disease or condition being treated.

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These additional agents may be administered separately from the proposed combination therapy, as part of a multiple dosing regimen. Alternatively, these agents may be part of a single dosage form, being mixed with a compound of the present invention into a single composition. When administered as part of a multiple dosing regimen, the two active agents may be administered simultaneously, sequentially, or within a period of time relative to each other, typically within five hours of each other.

As used herein, the terms combination, combination and related terms refer to the simultaneous or sequential administration of therapeutic agents in accordance with the present invention. For example, the combination of the present invention may be administered with another therapeutic agent simultaneously or sequentially in separate unit dosage forms or together in a unit dosage form. Accordingly, the present invention provides a single unit dosage form containing a compound of the present invention, an additional therapeutic agent and a pharmaceutically acceptable carrier, adjuvant or carrier. The amount of a compound of the present invention and the additional therapeutic agent (in compositions that contain the additional therapeutic agent as described above) that can be combined with the carrier materials to form a unit dosage form will vary depending on the host being treated and the particular route introduction. Preferably, the compositions of the present invention should be formulated such that a dose of 0.01-100 mg/kg body weight/day of a compound of the present invention can be administered.

In those compositions that contain an additional therapeutic agent, the additional therapeutic agent and the compound of the present invention may act synergistically. Thus, the amount of additional therapeutic agent in such compositions will be less than that required for monotherapy, when only one such therapeutic agent is used. In such compositions, said additional therapeutic agent may be administered in an amount of 0.01-1000 μg/kg body weight/day.

The amount of additional therapeutic agent present in the composition of the present invention will not exceed the amount that would normally be administered in a composition containing said therapeutic agent as the sole active agent. Preferably, the amount of additional therapeutic agent in the compositions disclosed herein will be from about 50 to 100% of the amount typically present in a composition containing such agent as the sole therapeutically active agent.

In one embodiment, the present invention provides a composition comprising a compound of Formula I and one or more additional therapeutic agents. Said therapeutic agent may be administered concomitantly with a compound of Formula I, or it may be administered before or after administration of a compound of Formula I. Suitable therapeutic agents are further described in detail below. In some embodiments, a compound of Formula I may be administered at intervals of up to 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours. , 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, or 18 hours before administration of the specified therapeutic agent. In other embodiments, a compound of Formula I may be administered at intervals of up to 5 minutes, 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours. , 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, 16 hours, 17 hours, or 18 hours after administration of the specified therapeutic agent.

In another embodiment, the present invention provides a method of treating a TCR-Nck mediated disease, disorder or condition by administering to a patient in need thereof a compound of Formula I and one or more additional therapeutic agents. Such additional therapeutic agents may include small molecules or recombinant biological agents and include, for example, acetaminophen, nonsteroidal anti-inflammatory drugs (NSAIDs) such as aspirin, ibuprofen, naproxen, etodolac (Lodine®) and celecoxib, colchicine (Colcrys®), corticosteroids , such as prednisone, prednisolone, methylprednisolone, hydrocortisone and the like, probenecid, allopurinol, febuxostat (Uloric®), sulfasalazine (Azulfidine®), antimalarials such as hydroxychloroquine (Plaquenil®) and chloroquine (Aralen®), methotrexate (Rheumatrex ®), gold salts such as aurothioglucose (Solganal®), aurothiomalate (Myochrysine®) and auranofin (Ridaura®), D-penicillamine (Depen® or Cuprimine®), azathioprine (Imuran®), cyclophosphamide (Cytoxan®), chlorambucil (Leukeran®), cyclosporine (Sandimmune®), leflunomide (Arava®), and anti-TNF agents such as etanercept (Enbrel®), infliximab (Remicade®), golimumab (Simponi®), certolizumab pegol (Cimzia®), and adalimumab ( Humira®), anti-IL-1 agents such as anakinra (Kineret®) and rilonacept (Arcalyst®), canakinumab (Ilaris®), Janus kinase inhibitors (anti-Jak) such as tofacitinib, antibodies such as rituximab (Rituxan®), anti-T cell agents such as abatacept (Orencia®), anti-IL-6 agents such as tocilizumab (Actemra®), diclofenac, cortisone, hyaluronic acid

- 31 045073 slots (Synvisc® or Hyalgan®), monoclonal antibodies such as tanezumab, anticoagulants such as heparin (Calcinparine® or Liquaemin®) and warfarin (Coumadin®), antidiarrheals such as diphenoxylate (Lomotil®) and loperamide (Imodium®), bile acid binding agents such as cholestyramine, alosetron (Lotronex®), lubiprostone (Amitiza®), laxatives such as magnesium hydroxide, polyethylene glycol (MiraLax®), Dulcolax®, Correctol® and Senokot®, anticholinergics or antispasmodics such as dicyclomine (Bentyl®), Singulair®, beta-2 agonists such as albuterol (Ventolin® HFA, Proventil® HFA), levosalbuterol (Xopenex®), metaproterenol (Alupent®), pirbuterol acetate (Maxair ®), terbutaline sulfate (Brethaire®), salmeterol xinafoate (Serevent®) and formoterol (Foradil®), anticholinergic agents such as ipratropium bromide (Atrovent®) and tiotropium (Spiriva®), inhaled corticosteroids such as beclomethasone dipropionate (Beclovent®) ®, Qvar® and Vanceril®), triamcinolone acetonide (Azmacort®), mometasone (Asthmanex®), budesonide (Pulmicort®) and flunisolide (Aerobid®), Afviar®, Symbicort®, Dulera®, sodium cromoglycate (Intal®), methylxanthines such as theophylline (Theo-Dur®, Theolair®, Slo-bid®, Uniphyl®, Theo24®) and aminophylline, IgE antibodies such as omalizumab (Xolair®), nucleoside reverse transcriptase inhibitors such as zidovudine (Retrovir® ), abacavir (Ziagen®), abacavir/lamivudine (Epzicom®), abacavir/lamivudine/zidovudine (Trizivir®), didanosine (Videx®), emtricitabine (Emtriva®), lamivudine (Epivir®), lamivudine/zidovudine (Combivir® ), stavudine (Zerit®) and zalcitabine (Hivid®), non-nucleoside reverse transcriptase inhibitors such as delavirdine (Rescriptor®), efavirenz (Sustiva®), nevirapine (Viramune®) and etravirine (Intelence®), nucleotide reverse transcriptase inhibitors, such as tenofovir (Viread®), protease inhibitors such as amprenavir (Agenerase®), atazanavir (Reyataz®), darunavir (Prezista®), fosamprenavir (Lexiva®), indinavir (Crixivan®), lopinavir and ritonavir (Kaletra®) , nelfinavir (Viracept®), ritonavir (Norvir®), saquinavir (Fortovase® or Invirase®) and tipranavir (Aptivus®), fusion inhibitors such as enfuvirtide (Fuzeon®) and maraviroc (Selzentry®), integrase inhibitors such as raltegravir (Isentress®), doxorubicin (Hydrodaunorubicin®), vincristine (Oncovin®), bortezomib (Velcade®), and dexamethasone (Decadron®) in combination with lenalidomide (Revlimid®), or any combination(s) thereof. In another embodiment, the present invention provides a method of treating rheumatoid arthritis, comprising administering to a patient in need thereof a compound of formula I and one or more additional therapeutic agents selected from nonsteroidal anti-inflammatory drugs (NSAIDs) such as aspirin, ibuprofen, naproxen, etodolac (Lodine ®) and celecoxib, corticosteroids such as prednisone, prednisolone, methylprednisolone, hydrocortisone and the like, sulfasalazine (Azulfidine®), antimalarials such as hydroxychloroquine (Plaquenil®) and chloroquine (Aralen®), methotrexate (Rheumatrex®), salts gold such as aurothioglucose (Solganal®), aurothiomalate (Myochrysine®) and auranofin (Ridaura®), D-penicillamine (Depen® or Cuprimine®), azathioprine (Imuran®), cyclophosphamide (Cytoxan®), chlorambucil (Leukeran®) , cyclosporine (Sandimmune®), leflunomide (Arava®) and anti-TNF agents such as etanercept (Enbrel®), infliximab (Remicade®), golimumab (Simponi®), certolizumab pegol (Cimzia®) and adalimumab (Humira®), anti-IL-1 agents such as anakinra (Kineret®) and rilonacept (Arcalyst®), antibodies such as rituximab (Rituxan®), anti-T cell agents such as abatacept (Orencia®), and anti- IL-6 agents such as tocilizumab (Actemra®).

In some embodiments, the present invention provides a method of treating osteoarthritis, comprising administering to a patient in need thereof a compound of formula I and one or more additional therapeutic agents selected from acetaminophen, nonsteroidal anti-inflammatory drugs (NSAIDs), such as aspirin, ibuprofen, naproxen, etodolac ( Lodine®) and celecoxib, diclofenac, cortisone, hyaluronic acid (Synvisc® or Hyalgan®) and monoclonal antibodies such as tanezumab.

In some embodiments, the present invention provides a method of treating systemic lupus erythematosus, comprising administering to a patient in need thereof a compound of Formula I and one or more additional therapeutic agents selected from acetaminophen, nonsteroidal anti-inflammatory drugs (NSAIDs), such as aspirin, ibuprofen, naproxen, etodolac (Lodine®) and celecoxib, corticosteroids such as prednisone, prednisolone, methylprednisolone, hydrocortisone and the like, antimalarials such as hydroxychloroquine (Plaquenil®) and chloroquine (Aralen®), cyclophosphamide (Cytoxan®), methotrexate (Rheumatrex® ), azathioprine (Imuran®), and anticoagulants such as heparin (Calcinparine® or Liquaemin®) and warfarin (Coumadin®).

In some embodiments, the present invention provides a method of treating Crohn's disease, ulcerative colitis, or inflammatory bowel disease, comprising administering to a patient in need thereof a compound of Formula I and one or more additional therapeutic agents selected from mesalamine (Asacol®), sulfasalazine (Azulfidine®), antidiarrheals such as diphenoxylate (Lomotil®) and loperamide (Imodium®), bile acid binders such as cholestyramine, alosetron (Lotronex®), lubiprostone (Amitiza®), laxatives such as

- 32 045073 as magnesium hydroxide, polyethylene glycol (MiraLax®), Dulcolax®, Correctol® and Senokot®, and anticholinergics or antispasmodics such as dicyclomine (Bentyl®), anti-TNF drugs, steroids and antibiotics such as Flagyl ( Flagyl®) or ciprofloxacin.

In some embodiments, the present invention provides a method of treating asthma, comprising administering to a patient in need thereof a compound of Formula I and one or more additional therapeutic agents selected from Singulair®, beta-2 agonists, such as albuterol (Ventolin® HFA, Proventil® HFA ), levosalbuterol (Xopenex®), metaproterenol (Alupent®), pirbuterol acetate (Maxair®), terbutaline sulfate (Brethaire®), salmeterol xinafoate (Serevent®) and formoterol (Foradil®), anticholinergic agents such as ipratropium bromide (Atrovent ®) and tiotropium (Spiriva®), inhaled corticosteroids such as prednisone, prednisolone, beclomethasone dipropionate (Beclovent®, Qvar® and Vanceril®), triamcinolone acetonide (Azmacort®), mometasone (Asthmanex®), budesonide (Pulmicort®), flunisolide (Aerobid®), Afviar®, Symbicort® and Dulera®, sodium cromoglycate (Intal®), methylxanthines such as theophylline (Theo-Dur®, Theolair®, Slo-bid®, Uniphyl®, Theo24®) and aminophylline, and IgE antibodies such as omalizumab (Xolair®).

In another embodiment, the present invention provides a method of treating a hematologic malignancy, comprising administering to a patient in need thereof a compound of formula I and one or more additional therapeutic agents selected from rituximab (Rituxan®), cyclophosphamide (Cytoxan®), doxorubicin (Hydrodaunorubicin®), vincristine (Oncovin®), prednisone, hedgehog signaling pathway inhibitor, BTK inhibitor, JAK/panJAK inhibitor, PI3K inhibitor, SYK inhibitor and combinations thereof. In another embodiment, the present invention provides a method of treating or reducing the severity of a disease, comprising administering to a patient in need thereof a compound of formula I and a BTK inhibitor, the disease being selected from inflammatory bowel disease, arthritis, systemic lupus erythematosus (SLE), vasculitis, idiopathic thrombocytopenic purpura (ITP), rheumatoid arthritis, psoriatic arthritis, osteoarthritis, Still's disease, juvenile arthritis, diabetes, myasthenia gravis, Hashimoto's thyroiditis, Ord's thyroiditis (an atrophic form of autoimmune thyroiditis), Graves' disease, autoimmune thyroiditis, Sjögren's syndrome, multiple sclerosis, systemic sclerosis, Lyme neuroborreliosis, Guillain-Barré syndrome, acute disseminated encephalomyelitis, Addison's disease, opsoclonus-myoclonus syndrome, ankylosing spondylosis, antiphospholipid syndrome, aplastic anemia, autoimmune hepatitis, autoimmune gastritis, pernicious anemia, celiac disease, Goodpasture's syndrome, idiopathic thrombocytopenic puritis puras, optic neuritis , scleroderma, primary biliary cirrhosis, Reiter's syndrome, Takayasu's arteritis, temporal arteritis, autoimmune hemolytic anemia with warm agglutinin syndrome, Wegener's granulomatosis, psoriasis, alopecia universalis, Behçet's disease, chronic fatigue, autonomic dystonia, membranous glomerulonephropathy, endometriosis, inter sticial cystitis, ordinary pemphigus, bullous pemphigus, neuromyotonia, scleroderma, vulvodynia, hyperproliferative disease, organ or tissue transplant rejection, acquired immunodeficiency syndrome (AIDS, also known as HIV), type 1 diabetes, graft-versus-host disease, transplantation, transfusion, anaphylaxis, allergies (eg , allergies to pollen, latex, drugs, food, insect poisons, animal dander, animal dander, dust mites or cockroach calix), type I hypersensitivity, allergic conjunctivitis, allergic rhinitis and atopic dermatitis, asthma, appendicitis, atopic dermatitis, asthma, allergies, blepharitis, bronchiolitis, bronchitis, bursitis, cervicitis, cholangitis, cholecystitis, chronic transplant rejection, colitis, conjunctivitis, Crohn's disease, cystitis, dacryoadenitis, dermatitis, dermatomyositis, encephalitis, endocarditis, endometritis, enteritis, enterocolitis, epicondylitis, epi didymite , fasciitis, fibrositis, gastritis, gastroenteritis, Henoch-Schönlein purpura, hepatitis, hidradenitis suppurativa, IgA nephropathy, interstitial lung disease, laryngitis, mastitis, meningitis, myelitis, myocarditis, myositis, nephritis, oophoritis, orchitis, osteitis, otitis, pancreatitis , mumps, pericarditis, peritonitis, pharyngitis, pleurisy, phlebitis, pneumonitis, pneumonia, polymyositis, proctitis, prostatitis, pyelonephritis, rhinitis, salpingitis, sinusitis, stomatitis, synovitis, tendonitis, tonsillitis, ulcerative colitis, uveitis, vaginitis, vasculitis or vulvitis, B-cell proliferative disorder, eg, diffuse large B-cell lymphoma, follicular lymphoma, chronic lymphocytic lymphoma, chronic lymphocytic leukemia, acute lymphocytic leukemia, B-cell prolymphocytic leukemia, lymphoplasmacytic lymphoma/Waldenström's macroglobulinemia, lymphoma s of the marginal zone of the spleen, multiple myeloma (also known like plaslet myeloma), non-podkhkinsky lymphoma, Hodgzhkin lymphoma, plasmocytoma, extranodal B-cell lymphoma of the marginal zone, nodal v-cell lymphoma of the marginal zone, mantle-cell lymphoma, large-cell-cell lymphoma (timus), intravascular coarse cereal-cereal. V-cell lymphoma, primary effusion lymphoma, Burkitt's lymphoma/leukemia or lymphomatoid granulomatosis, breast cancer, prostate cancer, or mast cell cancer (eg, mastocytoma, mast cell leukemia, mast cell sarcoma,

- 33 045073 systemic mastocytosis), bone cancer, colorectal cancer, pancreatic cancer, bone and joint diseases including, but not limited to, rheumatoid arthritis, seronegative spondyloarthropathy (including ankylosing spondylitis, psoriatic arthritis and Reiter's disease), Behçet's disease, Sjögren's syndrome, systemic sclerosis, osteoporosis, bone cancer, bone metastases, thromboembolic disorder (eg, myocardial infarction, angina pectoris, reocclusion after angioplasty, restenosis after angioplasty, reocclusion after coronary artery bypass grafting, restenosis after coronary artery bypass grafting, stroke, transient ischemia, peripheral arterial occlusive disease, pulmonary embolism, deep vein thrombosis), pelvic inflammatory disease, urethritis, skin sunburn, sinusitis, pneumonitis, encephalitis, meningitis, myocarditis, nephritis, osteomyelitis, myositis, hepatitis, gastritis, enteritis, dermatitis, gingivitis, appendicitis, pancreatitis, cholecystitis, agammaglobulinemia, psoriasis, allergies, Crohn's disease, irritable bowel syndrome, ulcerative colitis, Sjögren's disease, tissue transplant rejection, hyperacute organ transplant rejection, asthma, allergic rhinitis, chronic obstructive pulmonary disease (COPD), autoimmune polyglandular disease (also known as autoimmune polyglandular disease) syndrome), autoimmune alopecia, pernicious anemia, glomerulonephritis, dermatomyositis, multiple sclerosis, scleroderma, vasculitis, autoimmune hemolytic and thrombocytopenic conditions, Goodpasture's syndrome, atherosclerosis, Addison's disease, Parkinson's disease, Alzheimer's disease, diabetes, septic shock, systemic lupus erythematosus (SLE ), rheumatoid arthritis, psoriatic arthritis, juvenile arthritis, osteoarthritis, chronic idiopathic thrombocytopenic purpura, Waldenström's macroglobulinemia, myasthenia gravis, Hashimoto's thyroiditis, atopic dermatitis, degenerative joint disease, vitiligo, autoimmune hypopituitarism, Guillain-Barre syndrome, Behçet's disease, scleroderma, mushroom mycosis, acute inflammatory reactions (such as acute respiratory distress syndrome and ischemia/reperfusion injury) and Graves' disease.

In another embodiment, the present invention provides a method of treating or reducing the severity of a disease, comprising administering to a patient in need thereof a compound of formula I and a PI3K inhibitor, the disease being selected from cancer, a neurodegenerative disorder, an angiogenic disorder, a viral disease, an autoimmune disease, an inflammatory disorder, with hormone disease, conditions associated with organ transplantation, immunodeficiency disorders, destructive bone disease, proliferative disorder, infectious disease, condition associated with cell death, thrombin-induced platelet aggregation, chronic myelogenous leukemia (CML), chronic lymphocytic leukemia (CLL) , liver disease, pathological immune conditions accompanied by activation of T cells, cardiovascular disorders and central nervous system disorders.

In some embodiments, the present invention provides a method of treating or reducing the severity of a disease, comprising administering to a patient in need thereof a compound of Formula I and a Bcl-2 inhibitor, wherein the disease is an inflammatory disorder, an autoimmune disorder, a proliferative disorder, an endocrine disorder, a neurological disorder, or related to transplantation.

The compound of the present invention can also be used to obtain benefits in combination with antiproliferative compounds. Such antiproliferative compounds include, but are not limited to, aromatase inhibitors; antiestrogens; topoisomerase I inhibitors; topoisomerase II inhibitors; compounds having activity against microtubules; alkylating compounds; histone deacetylase inhibitors; compounds that induce cell differentiation processes; cyclooxygenase inhibitors; MMP inhibitors; mTOR inhibitors; antitumor antimetabolites; platinum compounds; compounds targeting/reducing protein kinase or lipid kinase activity and other antiangiogenic compounds; compounds that target, reduce or inhibit the activity of protein phosphatase or lipid phosphatase; gonadorelin agonists; antiandrogens; methionine aminopeptidase inhibitors; matrix metalloproteinase inhibitors; bisphosphonates; biological response modifiers; antiproliferative antibodies; heparanase inhibitors; inhibitors of oncogenic Ras isoforms; telomerase inhibitors; proteasome inhibitors; compounds used in the treatment of hematological malignancies; compounds that target, reduce or inhibit Flt-3 activity; Hsp90 inhibitors such as 17-AAG (17-allylaminogeldanamycin, NSC330507), 17-DMAG (17dimethylaminoethylamino-17-demethoxygeldanamycin, NSC707545), IPI-504, CNF1010, CNF2024, CNF1010 from Conforma Therapeutics; temozolomide (Temodal®); kinesin spindle protein inhibitors such as SB715992 or SB743921 from GlaxoSmithKline, or pentamidine/chlorpromazine from CombinatoRx; MEK inhibitors such as ARRY142886 from Array BioPharma, AZD6244 from AstraZeneca, PD181461 from Pfizer and leucovorin. The term aromatase inhibitor as used herein refers to a compound that inhibits the production of estrogens, eg, the conversion of the substrates androstenedione and testosterone to estrone and estradiol, respectively. The term includes, but is not limited to, steroids, in particular atamestane, exemestane and formestane and, in particular, non-steroidal compounds, in

- 34 045073 in particular aminoglutethimide, rogletimide, pyridoglutethimide, trilostane, testolactone, ketoconazole, vorozole, fadrozole, anastrozole and letrozole. Exemestane is marketed under the trade name Aromasin™. Formestane is marketed under the trade name Lentaron™. Fadrozole is marketed under the trade name Afema™. Anastrozole is marketed under the trade name Arimidex™. Letrozole is marketed under the trade names Femara™ or Femar™. Aminoglutethimide is marketed under the trade name Orimeten™. The combination according to the present invention containing a chemotherapeutic agent that is an aromatase inhibitor is particularly suitable for the treatment of hormone receptor-positive tumors such as breast tumors.

The term antiestrogen as used herein refers to a compound that antagonizes the effect of estrogens at the estrogen receptor level. The term includes, but is not limited to, tamoxifen, fulvestrant, raloxifene and raloxifene hydrochloride. Tamoxifen is marketed under the trade name Nolvadex™. Raloxifene hydrochloride is marketed under the trade name Evista™. Fulvestrant is available for administration under the trade name Faslodex™.

The term antiandrogen as used herein refers to any substance that is capable of inhibiting the biological effects of androgenic hormones and includes, but is not limited to, bicalutamide (Casodex™). As used herein, the term gonadorelin agonist includes, but is not limited to, abarelix, goserelin, and goserelin acetate. Goserelin is available for administration under the trade name Zoladex™.

The term topoisomerase I inhibitor as used herein includes, but is not limited to, topotecan, hymatecan, irinotecan, camptothecin and analogs thereof, 9-nitrocamptothecin and the macromolecular camptothecin conjugate PNU-166148. Irinotecan is available for administration, for example, in the form in which it is marketed, for example, under the brand name Camptosar™. Topotecan is marketed under the trade name Hycamptin™.

The term topoisomerase II inhibitor as used herein includes, but is not limited to, anthracyclines such as doxorubicin (including a liposomal formulation such as Caelyx™), daunorubicin, epirubicin, idarubicin and nerubicin, anthraquinones, mitoxantrone and losoxantrone, and the podophyllotoxins etoposide and teniposide. Etoposide is marketed under the trade name Etopophos™. Teniposide is marketed under the trade name VM 26-Bristol. Doxorubicin is marketed under the trade name Acriblastin™ or Adriamycin™. Epirubicin is marketed under the trade name Farmorubicin™. Idarubicin is marketed under the trade name Zavedos™. Mitoxantrone is marketed under the trade name Novantron.

The term microtubule-active agent refers to microtubule-stabilizing, microtubule-destabilizing, and microtubule polymerization inhibitor compounds, including, but not limited to, taxanes such as paclitaxel and docetaxel; vinca alkaloids such as vinblastine or vinblastine sulfate, vincristine or vincristine sulfate and vinorelbine; discodermolides; colchicine and epothilones and their derivatives. Paclitaxel is marketed under the trade name Taxol™. Docetaxel is marketed under the trade name Taxotere™. Vinblastine sulfate is marketed under the trade name Vinblastin R.P™. Vincristine sulfate is marketed under the trade name Farmistin™.

The term alkylating agent as used herein includes, but is not limited to, cyclophosphamide, ifosfamide, melphalan or nitrosourea (BCNU or Gliadel). Cyclophosphamide is marketed under the trade name Cyclostin™. Ifosfamide is marketed under the trade name Holoxan™. The term histone deacetylase inhibitors or HDAC inhibitors as used herein refers to compounds that inhibit histone deacetylase and that have antiproliferative activity. These include, but are not limited to, suberoylanilide hydroxamic acid (SAHA). The term antitumor antimetabolite as used herein includes, but is not limited to, 5-fluorouracil or 5-FU, capecitabine, gemcitabine, DNA demethylating compounds such as 5-azacytidine and decitabine, methotrexate and edatrexate, and folate antagonists such as pemetrexed. Capecitabine is marketed under the trade name Xeloda™. Gemcitabine is marketed under the trade name Gemzar™.

The term platinum compound as used herein includes, but is not limited to, carboplatin, cisplatin, cisplatin and oxaliplatin. Carboplatin can be administered, for example, in the form in which it is marketed, for example, under the trade name Carboplat™. Oxaliplatin is available for administration, for example, in the form in which it is marketed, for example, under the trade name Eloxatin™. As used herein, the term compounds that target/reduce protein kinase or lipid kinase activity; or protein phosphatase or lipid phosphatase activity; or other antiangiogenic compounds, includes, but is not limited to, protein tyrosine kinase inhibitors, and/or serine and/or threonine kinase inhibitors or lipid kinase inhibitors, such as a) compounds that target, reduce or inhibit the activity of platelet receptors

- 35 045073 growth factor (PDGFR), such as compounds that target, reduce or inhibit the activity of PDGFR, in particular, compounds that inhibit the TRF receptor, such as an N-phenyl-2-pyrimidinamine derivative such as imatinib, SU101, SU6668 and GFB-111; b) compounds that target, reduce or inhibit the activity of fibroblast growth factor receptors (FGFR); c) compounds that target, reduce or inhibit insulin-like growth factor receptor I (IGF-IR) activity, such as compounds that target, reduce or inhibit IGF-IR activity, in particular compounds that inhibit IGF receptor I kinase activity , or antibodies that target the extracellular domain of IGF receptor I or its growth factors; d) compounds that target, reduce or inhibit the activity of members of the Trk receptor tyrosine kinase family, or ephrin B4 inhibitors; f) compounds that target, reduce or inhibit the activity of members of the Axl family of receptor tyrosine kinases; f) compounds that target, reduce or inhibit the activity of Ret receptor tyrosine kinase; g) compounds that target, reduce or inhibit Kit/SCFR receptor tyrosine kinase activity, such as imatinib; h) compounds that target, reduce or inhibit the activity of C-kit receptor tyrosine kinases that are part of the PDGFR family, such as compounds that target, reduce or inhibit the activity of members of the c-Kit receptor tyrosine kinase family, in particular compounds that inhibit c-Kit receptor, such as imatinib; i) compounds that target, reduce or inhibit the activity of c-Abl family members, their gene fusion products (e.g. BCR-Abl kinase) and mutant variants, such as compounds that target, reduce or inhibit the activity of c-Abl family members and products fusions of their genes, such as an N-phenyl-2-pyrimidinamine derivative such as imatinib or nilotinib (AMN107); PD180970; AG957; NSC 680410; PD173955 from ParkeDavis; or dasatinib (BMS-354825); j) compounds that target, reduce or inhibit the activity of members of the protein kinase C (PKC) family and the Raf family of serine/threonine kinases, members of the MEK, SRC, JAK/pan-JAK, FAK, PDK1, PKB/Akt, Ras/MAPK, PI3K families , SYK, TYK2, BTK and TEC and/or members of the cyclin-dependent kinase (CDK) family, including staurosporine derivatives such as midostaurin; examples of other compounds include UCN-01, safingol, BAY 43-9006, bryostatin 1, perifosine; ilmofosine; RO 318220 and RO 320432; GO 6976; Isis 3521; LY333531/LY379196; isoquinoline compounds; FTI; PD184352 or QAN697 (PI3K inhibitor) or AT7519 (CDK inhibitor); k) compounds that target, reduce or inhibit the activity of protein tyrosine kinase inhibitors, such as compounds that target, reduce or inhibit the activity of protein tyrosine kinase inhibitors, including imatinib mesylate (Gleevec™) or tyrphostin, such as Tyrphostin A23/RG-50810; AG 99; Tyrphostin AG 213; Tyrphostin AG 1748; Tyrphostin AG 490; Tyrphostin B44; (+) enantiomer of Tyrphostin B44; Tyrphostin AG 555; AG 494; Tyrphostin AG 556, AG957 and adaphostin, adamantyl ester (4-{[(2,5-dihydroxyphenyl)methyl]amino}benzoic acid; NSC 680410, adaphostin); l) compounds that target, reduce or inhibit the activity of members of the epidermal growth factor receptor tyrosine kinase family (EGFR1 ErbB2, ErbB3, ErbB4 as homo- or heterodimers) and mutant variants thereof, such as compounds that target, reduce or inhibit the activity of members the epidermal growth factor (EGF) receptor family, in particular, compounds, proteins or antibodies that inhibit members of the EGF receptor tyrosine kinase family, such as EGF receptor, ErbB2, ErbB3 and ErbB4, or bind to EGF or EGF-related ligands, CP 358774, ZD 1839, ZM 105180.; trastuzumab (Herceptin™), cetuximab (Erbitux™), Iressa, Tarceva, OSI-774, CI-1033, EKB-569, GW-2016, E1.1, E2.4, E2.5, E6.2, E6. 4, E2.11, E6.3 or E7.6.3 and 7H-pyrrolo[2,3-d]pyrimidine derivatives; m) compounds that target, reduce or inhibit the activity of the c-Met receptor, such as compounds that target, reduce or inhibit the activity of c-Met, in particular compounds that inhibit the kinase activity of the c-Met receptor or antibodies that target the extracellular domain c-Met or HGF binding, n) compounds that target, reduce or inhibit the kinase activity of one or more members of the JAK family (JAK1/JAK2/JAK3/TYK2 and/or pan-JAK), including, but not limited to, PRT- 062070, SB-1578, baricitinib, pacritinib, momelotinib, VX-509, AZD-1480, TG-101348, tofacitinib and ruxolitinib; o) compounds aimed at reducing or inhibiting the kinase activity of PI3 kinase (PI3K), including, but not limited to, ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, buparlisib, pictrelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765 and idelalisib; And; and q) compounds that target, reduce or inhibit the signaling effects of hedgehog (Hh) or smoothed receptor (SMO) protein pathways, including, but not limited to, cyclopamine, vismodegib, itraconazole, erizmodegib and IPI-926 (saridegib).

The term PI3K inhibitor as used herein includes, but is not limited to, compounds having inhibitory activity against one or more enzymes of the phosphatidylinositol 3-kinase family, including, but not limited to, PI3Ka, PI3Ky, PI3Kδ, PI3Ke, PI3KC2a, PI3K -C2e, PI3K-C2y, Vps34, p110-a, p110-v, p110-y, p110-δ, p85-a, p85-v, p55-y, p150, p101 and p87. Examples of PI3K inhibitors that may be used in the present invention include, but are not limited to:

- 36 045073 are limited to them, ATU-027, SF-1126, DS-7423, RBI-05204, GSK-2126458, ZSTK-474, buparlisib, picterlisib, PF-4691502, BYL-719, dactolisib, XL-147, XL- 765 and idelalisib. The term Bcl2 inhibitor as used herein includes, but is not limited to, compounds having inhibitory activity against B-cell lymphoma protein 2 (Bcl-2), including, but not limited to, ABT-199, ABT-731, ABT-737 , apogosipol, pan-Blc-2 inhibitors Ascenta, curcumin (and its analogs), dual-acting Bcl-2/Bcl-xL inhibitors (Infinity Pharmaceuticals/Novartis Pharmaceuticals), Genasense (G3139), HA14-1 (and its analogs; see WO 2008/118802), navitoclax (and its analogs, see US 7390799), NH1 (Shenayng Pharmaceutical University), obatoclax (and its analogs, see WO 2004/106328), S-001 (Gloria Pharmaceuticals), compounds TW series (Univ. of Michigan) and venetoclax. In some embodiments, the Bcl-2 inhibitor is a small molecule drug. In some embodiments, the Bcl-2 inhibitor is a peptidomimetic. The term BTK inhibitor as used herein includes, but is not limited to, compounds having Bruton's tyrosine kinase (BTK) inhibitory activity, including, but not limited to, AVL-292 and ibrutinib. The term SYK inhibitor as used herein includes, but is not limited to, compounds having spleen tyrosine kinase (SYK) inhibitory activity, including, but not limited to, PRT-062070, R-343, R-333, Excellair, PRT-062607 and fostamatinib.

Other examples of BTK inhibitory compounds and conditions that can be treated with such compounds in combination with the compounds of the present invention can be found in WO 2008/039218 and WO 2011/090760, the entire contents of which are incorporated herein by reference.

Other examples of SYK inhibitory compounds and conditions that can be treated with such compounds in combination with the compounds of the present invention can be found in WO 2003/063794, WO 2005/007623 and WO 2006/078846, the entire contents of which are incorporated herein by reference. .

Other examples of PI3K inhibitory compounds and conditions that can be treated with such compounds in combination with compounds of the present invention can be found in WO 2004/019973, WO 2004/089925, WO 2007/016176, US 8138347, WO 2002/088112, WO 2007 /084786, WO 2007/129161, WO 2006/122806, WO 2005/113554 and WO 2007/044729, the entire contents of which are incorporated herein by reference.

Other examples of JAK inhibitory compounds and conditions that can be treated with such compounds in combination with the compounds of the present invention can be found in WO 2009/114512, WO 2008/109943, WO 2007/053452, WO 2000/142246 and WO 2007/070514. the entire contents of said references are incorporated herein by reference.

Others include compounds whose activity is mediated by a mechanism other than protein kinase or lipid kinase inhibition, such as Thalomid™ and TNP-470.

Examples of proteasome inhibitors suitable for use in combination with the compounds of the present invention include, but are not limited to, bortezomib, disulfiram, epigallocatechin-3-gallate (EGCG), salinosporamide A, carfilzomib, ONX-0912, CEP-18770 and MLN9708.

Compounds that target, reduce or inhibit the activity of protein phosphatase or lipid phosphatase are, for example, phosphatase 1, phosphatase 2A or CDC25 inhibitors, such as okadaic acid or a derivative thereof.

Compounds that induce cell differentiation processes include, but are not limited to, retinoic acid, α-γ- or δ-tocopherol, or α-γ-or δ-tocotrienol.

The term cyclooxygenase inhibitor as used herein includes, but is not limited to, Cox-2 inhibitors, 5-alkyl-2-arylaminophenylacetic acid and derivatives thereof, such as celecoxib (Celebrex™), rofecoxib (Vioxx™), etoricoxib, valdecoxib or 5-alkyl -2-arylaminophenylacetic acid, such as 5-methyl-2-(2'-chloro-6'-fluoroanilino)phenylacetic acid, lumiracoxib. The term bisphosphonates as used herein includes, but is not limited to, etridonic acid, clodronic acid, tiludronic acid, pamidronic acid, alendronic acid, ibandronic acid, risedronic acid, and zoledronic acid. Etridonic acid is marketed under the trade name Didronel™. Clodronic acid is marketed under the trade name Bonefos™. Tiludronic acid is marketed under the trade name Skelid™. Pamidronic acid is marketed under the trade name Aredia™. Alendronic acid is marketed under the trade name Fosamax™. Ibandronic acid is marketed under the trade name Bondranat™. Risedronic acid is marketed under the trade name Actonel™. Zoledronic acid is marketed under the trade name Zometa™. The term mTOR inhibitors refers to compounds that inhibit mammalian target of rapamycin (mTOR) and that have antiproliferative activity, such as sirolimus (Rapamune®), everolimus (Certican™), CCI-779 and ABT578. The term heparanase inhibitor as used herein refers to compounds that target, reduce or inhibit the degradation of heparin sulfate. Term

- 37 045073 includes, but is not limited to, PI-88. The term biological response modifier as used herein refers to lymphokine or interferons.

The term inhibitor of oncogenic Ras isoforms, such as H-Ras, K-Ras or N-Ras, as used herein refers to compounds that target, reduce or inhibit the oncogenic activity of Ras; an example is a farnesyltransferase inhibitor such as L-744832, DK8G557 or R115777 (Zamestra™). As used herein, the term telomerase inhibitor refers to compounds that target, reduce or inhibit telomerase activity. Compounds that target, reduce or inhibit telomerase activity are, in particular, compounds that inhibit the telomerase receptor, such as telomestatin.

The term methionine aminopeptidase inhibitor as used herein refers to compounds that target, reduce or inhibit the activity of methionine aminopeptidase. Compounds that target, reduce or inhibit methionine aminopeptidase activity include, but are not limited to, benhamide or a derivative thereof.

The term proteasome inhibitor as used herein refers to compounds that target, reduce or inhibit proteasome activity. Compounds that target, reduce or inhibit proteasome activity include, but are not limited to, Bortezomib (Velcade™) and MLN 341. The term matrix metalloproteinase inhibitor or (MMP inhibitor) as used herein includes, but is not limited to, collagen inhibitors from class of peptidomimetics and non-peptidomimetics, tetracycline derivatives, for example, the hydroxamate inhibitor from the class of peptidomimetics, batimastat and its analogue, which is bioavailable when administered orally, marimastat (BB-2516), prinomastat (AG3340), metastat (NSC 683551), BMS-279251, BAY 129566, TAA211, MMI270B or AAJ996.

The term compounds used in the treatment of hematological malignancies as used herein includes, but is not limited to, FMS-like tyrosine kinase inhibitors, which are compounds that target, reduce or inhibit the activity of FMS-like tyrosine kinase receptors (Flt-3R); interferon, 1-β-Οarabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitors, which are compounds that target, reduce or inhibit anaplastic lymphoma kinase activity.

Compounds that target, reduce or inhibit the activity of FMS-like tyrosine kinase receptors (Flt-3R) are, in particular, compounds, proteins or antibodies that inhibit members of the Flt-3R receptor kinase family, such as PKC412, midostaurin, a staurosporine derivative, SU11248 and MLN518.

The term HSP90 inhibitors as used herein includes, but is not limited to, compounds that target, reduce or inhibit the ATPase activity inherent in HSP90; involved in the degradation of, targeting, reducing, or inhibiting HSP90-dependent proteins through the ubiquitinin-proteasome pathway. Compounds that target, reduce or inhibit the ATPase activity inherent in HSP90 are, in particular, compounds, proteins or antibodies that inhibit the ATPase activity of HSP90, such as 17allylamino,17-demethoxygeldanamycin (17AAG), a derivative of geldanamycin; other geldanamycin-related compounds; radicicol and HDAC inhibitors. The term antiproliferative antibodies as used herein includes, but is not limited to, trastuzumab (Herceptin™), Trastuzumab-DMI, Erbitux, bevacizumab (Avastin™), rituximab (Rituxan®), PRO64553 (anti-CD40) and 2C4. Antibodies include intact monoclonal antibodies, polyclonal antibodies, polyspecific antibodies derived from at least 2 intact antibodies, and antibody fragments, provided they exhibit the desired biological activity.

For the treatment of acute myeloid leukemia (AML), the compounds of the present invention can be used in combination with standard treatments for leukemia, in particular in combination with treatments used for the treatment of AML. In particular, the compounds of the present invention can be administered in combination with, for example, farnesyltransferase inhibitors and/or other drugs suitable for use in the treatment of AML, such as daunorubicin, adriamycin, Ara-C, VP-16, teniposide, mitoxantrone, idarubicin , carboplatin and PKC412.

Other antileukemic compounds include, for example, Ara-C, a pyrimidine analogue, which is a 2'-alpha-hydroxyribose (arabinoside) derivative of deoxycytidine. Also included are the purine analogue of hypoxanthine, 6-mercaptopurine (6-MP), and fludarabine phosphate. Compounds that target, reduce, or inhibit histone deacetylase (HDAC) activity, such as sodium butyrate and suberoylanilide hydroxamic acid (SAHA), inhibit the activity of enzymes known as histone deacetylases. Specific HDAC inhibitors include MS275, SAHA, FK228 (formerly FR901228), trichostatin A and the compounds disclosed in US Pat. No. 6,552,065, including, but not limited to, N-hydroxy-3-[4-[[[2-(2-methyl -1H-indol-3-yl)ethyl]amino]methyl]phenyl]-2E2-propenamide or a pharmaceutically acceptable salt thereof and N-hydroxy-3-[4-[(2-hydroxyethyl){2-(1Nindol-3- yl)ethyl]amino]methyl]phenyl]-2E-2-propenamide or a pharmaceutically acceptable salt thereof, in

- 38 045073 particularly lactate salt. As used herein, somatostatin receptor antagonists refer to compounds that target, act on, or inhibit the somatostatin receptor, such as octreotide and SOM230. Approaches based on tumor cell damage include approaches such as ionizing radiation. The term ionizing radiation, as referred to above and below, means ionizing radiation that occurs in the form of either electromagnetic radiation (such as X-rays and gamma rays) or particles (such as alpha and beta particles). Ionizing radiation has been proposed for, without limitation, radiation therapy and is known in the art. See Hellman, Principles of Radiation Therapy, Cancer, in Principles and Practice of Oncology, Devita et al., Eds., 4th Edition, Vol. 1, pp. 248-275 (1993).

Also included are EDG binders and ribonucleotide reductase inhibitors. The term EDG binders as used herein refers to a class of immunosuppressants that modulate lymphocyte recycling, such as FTY720. The term ribonucleotide reductase inhibitors refers to pyrimidine or purine nucleoside analogues, including, but not limited to, fludarabine and/or cytosine arabinoside (ara-C), 6-thioguanine, 5-fluorouracil, cladribine, 6-mercaptopurine (particularly in combination with ara -C against ALL) and/or pentostatin. Ribonucleotide reductase inhibitors are, in particular, hydroxyurea or 2hydroxy-1H-isoindole-1,3-dione derivatives. Also included, in particular, are compounds, proteins or monoclonal antibodies against vascular endothelial growth factor (VEGF) such as 1-(4-chloroanilino)-4-(4pyridylmethyl)phthalazine or its pharmaceutically acceptable salt, 1-(4-chloroanilino succinate )-4-(4pyridylmethyl)phthalazine; Angiostatin™; Endostatin™; anthranilic acid amides; ZD4190; ZD6474; SU5416; SU6668; bevacizumab; or anti-VEGF antibodies or anti-VEGF receptor antibodies such as rhuMAb and RHUFab, a VEGF aptamer such as Macugon; FLT-4 inhibitors, FLT-3 inhibitors, anti-VEGFR-2 IgG1 antibody, angiozyme (RPI 4610) and bevacizumab (Avastin™).

Photodynamic therapy as used herein refers to a therapy that uses certain chemicals known as photosensitizing compounds to treat or prevent cancer. Examples of photodynamic therapy include treatment with compounds such as Visudyne™ and sodium porfimer.

Angiostatic steroids herein refer to compounds that block or inhibit angiogenesis, such as, for example, anecortave, triamcinolone, hydrocortisone, 11α-epihydrocortisol, cortexolone, 17α-hydroxyprogesterone, corticosterone, deoxycorticosterone, testosterone, estrone and dexamethasone.

Implants containing corticosteroids include compounds such as fluocinolone and dexamethasone.

Other chemotherapeutic compounds include, but are not limited to, plant alkaloids, hormonal compounds and antagonists; biological response modifiers, preferably lymphokines or interferons; antisense oligonucleotides or derivatives of oligonucleotides; shRNA or siRNA; or other compounds or compounds with a different or unknown mechanism of action. The structure of active compounds was identified by code numbers, nonproprietary names or trademarks, which can be obtained from the current edition of the standard manual The Merck Index or from databases such as Patents International (eg, IMS World Publications).

The compound of the present invention can also be used in combination with known therapeutic processes, for example the administration of hormones or radiation. In some embodiments, the proposed compound is used as a radiosensitizer, in particular for the treatment of tumors that exhibit poor sensitivity to radiotherapy.

The compounds of the present invention or pharmaceutical compositions thereof may also be included in compositions for coating implantable medical devices such as prostheses, artificial valves, vascular grafts, stents and catheters. Vascular stents, for example, are used to avoid restenosis (re-narrowing of the vessel wall after injury). However, for patients using stents or other implantable devices, there is a risk of clot formation or platelet activation. These undesirable effects can be prevented or reduced by precoating the device with a pharmaceutically acceptable composition containing a kinase inhibitor. Implantable devices coated with the compound of the present invention are another embodiment of the present invention.

Examples

The following examples are intended to illustrate the present invention and should not be construed as limiting the present invention. Unless otherwise indicated, one or more tautomeric forms of the compounds according to the examples described herein can be prepared in situ and/or isolated. All tautomeric forms of the compounds according to the following examples are to be considered disclosed herein. Temperature values are given in degrees

- 39 045073

Celsius. Unless otherwise specified, all evaporation procedures are carried out under reduced pressure, preferably between about 15 mmHg. Art. and 100 mm Hg. Art. (= 20-133 mbar). The structures of the final products, intermediates and starting materials are confirmed by standard analytical methods, e.g. microanalysis and spectroscopic characterizations, e.g. MS, IR,

NMR. The abbreviations used are traditional in the art.

It will be appreciated that while the general methods reflect the synthesis of the individual compounds of the present invention, the following general methods and other methods known to one of ordinary skill in the art can be used to prepare all compounds, subclasses and species of each of these compounds as described herein .

Certain compounds described herein may also be prepared using synthetic methods described in earlier US patents, such as US Patent Nos. 8,614,231, 9,120,764, 10,131,647 and 10,106,518, each of which is incorporated herein by reference in its entirety.

List of abbreviations used in the experimental part:

4A MS: molecular sieves with pore size 4A,

AcOH: acetic acid,

Anhydrous: anhydrous,

Water: water,

BH ₃ -THF: complex of borane and tetrahydrofuran,

BINAP: (2,2'-bis(diphenylphosphino)-1,1'-binaphthyl),

Bn: benzyl,

Boc: tert-butoxycarbonyl, (Boc) ₂ O: di-tert-butyl dicarbonate,

BrettPhos: 2-(dicyclohexylphosphino)3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl,

CbzCl: benzyl chloroformate,

Cbz-OSU: N-(benzyloxycarbonyloxy)succinimide,

CHIRAL HPLC: chiral high performance liquid chromatography,

CMBP: (cyanomethylene)tributylphosphorane,

Conc.: concentrated,

CuCN: copper cyanide, day: day, DAST: diethylaminosulfur trifluoride, DavePhos: 2-dicyclohexylphosphino-2'-(C/M-dimethylamino)biphenyl, dba: dibenzylideneacetone,

DBU: 1,8-diazobicyclo[5.4.0]undec-7-ene,

EDC: 1,2-dichloroethane,

DCM: dichloromethane,

DEA: diethylamine,

DIBAL-H: diisobutylaluminum hydride,

DIPEA: N,N-diisopropylethylamine,

DMA: N,N-dimethylacetamide,

DMAP: 4-dimethylaminopyridine,

DMF: N,N-dimethylformamide,

DMPU: 1,3-dimethyl-3,4,5,6-tetrahydro-2-pyrimidinone,

DMSO: dimethyl sulfoxide,

DPPA: diphenylphosphoryl azide, dppf: 1,1'-bis(diphenylphosphino)ferrocene,

EA: ethyl acetate,

EDCI: 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride,

EDTA: ethylenediaminetetraacetic acid,

EI: enantiomeric excess,

ESI: electrospray ionization,

Et ₃ N: triethylamine,

Et ₂ O: diethyl ether,

EtOAc: ethyl acetate,

EtOH: ethanol,

Fmoc: fluorenylmethyloxycarbonyl,

Fmoc-OSu: N-(9-fluorenylmethoxycarbonyloxy)succinimide, h: hours,

HATU: N,N,N',N'-tetramethyl-O-(7-azabenzotriazol-1-yl)uranium hexafluorophosphate,

HOBt: Hydroxybenzotriazole,

HPLC: high performance liquid chromatography,

- 40 045073

HCl: hydrochloric acid,

IBX: 2-iodoxybenzoic acid,

IPA: isopropyl alcohol,

JackiePhos: 2-{bis[3,5-bis(trifluoromethyl)phenyl]phosphino}-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'biphenyl, bis(3,5-bis (trifluoromethyl)phenyl)(2',4',6'-triisopropyl-3,6-dimethoxybiphenyl-2-yl)phosphine,

LDA: lithium diisopropylamide, M: molar, mCPBA: meta-chloroperoxybenzoic acid, Me: methyl, MeCN: acetonitrile, MeOH: methanol, MgO: magnesium oxide, min: minutes, ml: milliliters, mM: millimolar, mmol: millimoles, MOM : methoxymethyl, MsCl: mesyl chloride, MTBE: methyl tert-butyl ether, NMP: N-methyl-2-pyrrolidone, n-BuLi: n-butyllithium, NBS: N-bromosuccinimide, NIS: N-iodosuccinimide, NMO: 4- methylmorpholine N-oxide, NMP: N-methylpyrrolidone, NMR: nuclear magnetic resonance, °C: degrees Celsius, PBS: phosphate buffered saline, Pd/C: palladium on carbon, Pd ₂ (dba) ₃ : tris(dibenzylideneacetone)dipalladium (0), PE: petroleum ether, prep-HPLC: preparative high performance liquid chromatography, P(o-tol) ₃ : tri(o-tolyl)phosphine, PTFE: polytetrafluoroethylene, Rel: relative, rt: room temperature ,

RuPhos: 2-dicyclohexylphosphino-2',6'-diisopropoxybiphenyl, us: saturated, SFC: supercritical fluid chromatography, SGC: silica gel chromatography, STAB: sodium triacetoxyborohydride, TBAB: tetra-n-butylammonium bromide, TBAF: tetra-n- butylammonium fluoride, TBSCl: tert-butyldimethylsilyl chloride, tBuOK: potassium tert-butoxide, tBuONa: sodium tert-butoxide, TEA: triethylamine, TEBAC: benzyltriethylammonium chloride, Tf: trifluoromethanesulfonate, TFUA: trifluoromethanesulfonic acid anhydride, TFA: trifluoroacetic acid, TIPS:triisopropylsilyl , TLC: thin layer chromatography, THF: tetrahydrofuran, TMSCN: trimethylsilyl cyanide, pTSA: p-toluenesulfonic acid, TsOH: p-toluenesulfonic acid, XantPhos: 4,5-bis(diphenylphosphino)-9,9-dimethylxanthene, XPhos: 2-dicyclohexylphosphino -2',4',6'-triisopropylbiphenyl. General methods for preparing the compounds of the present invention. The preparation of illustrative non-limiting examples of the proposed compounds is described below. The compounds of the present invention can be prepared or isolated according to

- 41 045073 with general synthetic and/or semi-synthetic methods known to those skilled in the art for similar compounds, and the methods described in detail in the Examples section of this specification.

In the diagrams below, when a particular protecting group (PG), leaving group (LG) or conversion condition is depicted, one of ordinary skill in the art will recognize that other protecting groups, leaving groups and conversion conditions are also suitable and the present invention is also included. Such groups and transformations are described in detail in March's Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, M. B. Smith and J. March, ^5th Edition, John Wiley & Sons, 2001, Comprehensive Organic Transformations, R. S. Larock, ^2nd Edition, John Wiley & Sons, 1999, and Protecting Groups in Organic Synthesis, TW Greene and PGM Wuts, ^3rd edition, John Wiley & Sons, 1999, the entire contents of each of these references are incorporated herein by reference.

As used herein, the phrase leaving group (LG) includes, but is not limited to, halogens (e.g., fluoride, chloride, bromide, iodide), sulfonates (e.g., mesylate, tosylate, benzenesulfonate, brosylate, nosylate, triflate), diazonium, and the like .

As used herein, the phrase oxygen protecting group includes, for example, carbonyl protecting groups, hydroxyl protecting groups, etc. Hydroxyl protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and PGM Wuts, ^3rd edition, John Wiley & Sons, 1999, the entire contents of which are incorporated herein by links. Examples of suitable hydroxyl protecting groups include, but are not limited to, esters, allyl ethers, ethers, silyl ethers, alkyl ethers, arylalkyl ethers and alkoxyalkyl ethers. Examples of such esters include formates, acetates, carbonates and sulfonates. Specific examples include formate, benzoyl formate, chloroacetate, trifluoroacetate, methoxyacetate, triphenylmethoxyacetate, p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate, 4,4(ethylenedithio)pentanoate, pivaloate (trimethylacetyl), crotonate, 4-methoxycrotonate, benzoate, pbenyl benzoate, 2,4,6-trimethylbenzoate, carbonates such as methyl, 9-fluorenylmethyl, ethyl, 2,2,2trichloroethyl, 2-(trimethylsilyl)ethyl, 2-(phenylsulfonyl)ethyl, vinyl, allyl and p-nitrobenzyl. Examples of such silyl ethers include trimethylsilyl, triethylsilyl, tbutyldimethylsilyl, t-butyldiphenylsilyl, triisopropylsilyl and other trialkylsilyl ethers. Alkyl ethers include methyl, benzyl, pmethoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, allyl and allyloxycarbonyl ethers or derivatives. Alkoxyalkyl ethers include acetals such as methoxymethyl, methylthiomethyl, (2-methoxyethoxy)methyl, benzyloxymethyl, beta(trimethylsilyl)ethoxymethyl, and tetrahydropyranyl ethers. Examples of arylalkyl ethers include benzyl, p-methoxybenzyl (MPM), 3,4-dimethoxybenzyl, onitrobenzyl, p-nitrobenzyl, p-halobenzyl, 2,6-dichlorobenzyl, pcyanobenzyl, and 2- and 4-picolyl ethers.

Amine protecting groups are well known in the art and include those described in detail in Protecting Groups in Organic Synthesis, T. W. Greene and PGM Wuts, ^3rd edition, John Wiley & Sons, 1999, the entire contents of which are incorporated herein. via link. Suitable amino protecting groups include, but are not limited to, aralkylamines, carbamates, cyclic imides, allylamines, amides and the like. Examples of such groups include t-butyloxycarbonyl (BOC), ethyloxycarbonyl, methyloxycarbonyl, trichloroethyloxycarbonyl, allyloxycarbonyl (Alloc), benzyloxocarbonyl (CBZ), allyl, phthalimide, benzyl (Bn), fluorenylmethylcarbonyl (Fmoc), formyl, acetyl, chloroacetyl, dichloroacetyl, trichloroacetyl , phenylacetyl, trifluoroacetyl, benzoyl and the like. All starting materials, building blocks, reagents, acids, bases, dehydrating agents, solvents and catalysts used for the synthesis of the compounds of the present invention are either commercially available or can be prepared by organic synthesis methods known to one of ordinary skill in the art. (Houben-Weyl 4th Ed. 1952, Methods of Organic Synthesis, Thieme, Volume 21). In addition, the compounds of the present invention can be prepared by organic synthesis methods known to one of ordinary skill in the art, as shown in the following examples.

One skilled in the art will appreciate that the various functional groups present in the compounds of the present invention, such as aliphatic groups, alcohols, carboxylic acids, esters, amides, aldehydes, halogens and nitriles, can be interconverted by methods well known in the art. in the art, including, but not limited to, reduction, oxidation, esterification, hydrolysis, partial oxidation, partial reduction, halogenation, dehydration, partial hydration and hydration. (See, for example, March's Advanced Organic Chemistry, ^5th Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001). Such interconversions may require the use of the above procedures, and some methods for synthesizing the compounds of the present invention are described below.

- 42 045073

Examples

Synthesis scheme 1.

Step 1: synthesis of 3-(4-(trifluoromethoxy)phenoxy)propanoic acid.

Chloropropanoic acid (24.49 g, 224.7 mmol) in 2N. NaOH (100 ml) was added dropwise over 10 min at 100°C to a well stirred mixture of 4-(trifluoromethoxy)phenol (compound 1; 20 g, 112.35 mmol). The reaction mixture was stirred for 4 hours at 100°C. After completion, the reaction mixture was cooled to room temperature and acidified to pH 2-3 using 1N. HCl. The resulting solid was collected by filtration to obtain 4 g of crude compound 2, which was directly used in the next step without any further purification.

Step 2: synthesis of 6-(trifluoromethoxy)chroman-4-one (compound 3).

Trifluoromethanesulfonic acid (2.14 ml) was added dropwise over 10 min to a well stirred mixture of compound 2 (4 g, 16.06 mmol) in TFA (4.56 g, 40.0 mmol) at 0°C. The reaction mixture was stirred for 24 hours. Once complete, the reaction mixture was poured into cold H ₂ O (100 ml) and extracted with EtOAc (2x150 ml). The combined organic layers were washed with H ₂ O (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography (hexane:EtOAc; 95:5) to give compound 3 (2.8 g).

Step 3: synthesis of 4-(4-fluorophenyl)-6-(trifluoromethoxy)-2H-chromene.

n-BuLi (4.9 mL, 1.6 M in hexane, 8.0 mmol) was added dropwise over 10 min at -78°C to a well-stirred mixture of 4-fluorobromobenzene (1.0 g, 8.0 mmol ) in THF (20 ml). The reaction mixture was stirred at -78°C for 30 minutes. Compound 3 (4.5 g, 30.4 mmol) was dissolved in THF (20 ml) and added dropwise to the reaction mixture over 10 min at -78°C. The reaction temperature was gradually increased to -50°C and stirred until the starting material was consumed by TLC.

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After 4 h, the reaction temperature was brought to -30°C. Once complete, aqueous NH4Cl (50 ml) was added to the reaction mixture and extracted with EtOAc (2x150 ml). The combined organic layers were washed with H ₂ O (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The organic layer was evaporated under reduced pressure to obtain a crude product which was directly used in the next step without any purification or analysis. To this crude product was added 20% aqueous H2SO4 (5 ml) and 1,4-dioxane (5 ml) and the reaction mixture was refluxed for 6 hours at 95°C, then the contents were washed with NaOH (1%, 20 ml ) and subjected to extraction with EtOAc (2x100 ml). The combined organic layers were washed with H ₂ O (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography (hexane) to give compound 4 (0.525 g).

Step 5: synthesis of 4-(4-fluorophenyl)-6-(trifluoromethoxy)-2H-chromene-3-carbaldehyde.

DMF (368 mg, 5.03 mmol) was added dropwise to POCl ₃ (770 mg, 5.03 mmol) in a round bottom flask and the contents were heated to 40°C for 45 min. A solution of compound 4 (520 mg, 1.677 mmol) in DMF (1 ml) was added dropwise to the same reaction flask and the contents were heated to 60°C for 15 hours. The reaction mixture was cooled to room temperature and diluted with saturated aqueous NaHCO ₃ ( pH ~ 8) and subjected to extraction with EtOAc (2x50 ml). The combined organic layers were washed with H2O (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography (hexane) to give compound 5 (0.455 g).

Step 6: synthesis of 4-(4-fluorophenyl)-6-(trifluoromethoxy)-2H-chromene-3-carboxylic acid.

NaH ₂ PO ₄ -2H ₂ O (551 mg, 3.99 mmol), NaClO ₂ (359 mg, 3.99 mmol) and 2-methyl-2-butene (932 mg, 13.3 mmol) were added to the well stirred a solution of compound 5 (450 mg, 1.33 mmol) in t-BuOH (25 ml), followed by water (2 ml) at room temperature. The reaction mixture was then stirred for 20 minutes. The reaction was monitored by TLC; upon completion, the contents were evaporated and the pH of the reaction was adjusted to a value of 2-3 by adding 2N. HCl. A precipitated solid obtained and was isolated by filtration and dried to give compound 6 (0.463 g).

Step 7: synthesis of N-(2-(dimethylαamino)ethyl)-4-(4-fluorophenyl)-6-(trifluoromethoxy)-2H-chromene-3carboxamide (I-10).

HOBt (175 mg, 1.29 mmol) was added to a mixture of compound 6 (460 mg, 1.29 mmol), diamine (138 mg, 1.95 mmol), EDC.HCl (374 mg, 1.95 mmol) and DIPEA (838 mg, 6.49 mmol) in THF (5 ml) at room temperature. The reaction mixture was then irradiated (microwave radiation) for 3 minutes. Once complete, the contents were poured into cold H2O (20 ml) and extracted with EtOAc (2x30 ml). The combined organic layers were washed with H ₂ O (20 ml) and brine (10 ml), and separated and then dried over anhydrous Na2SO4. The crude product was purified by preparative HPLC to give I-10 (31 mg).

Example 2: 1-((4-(4-fluorophenyl)-6-(trifluoromethoxy)-2H-chromen-3-yl)methyl)-4-methylpiperazine.

F

1-11

Step 1: synthesis of 3-(chloromethyl)-4-(4-fluorophenyl)-6-(trifluoromethoxy)-2H-chromene.

Sodium borohydride (0.5 eq.) in toluene (20 ml) was added dropwise at room temperature to a mixture of compound 5 (3 g, 1 eq.) in methanol (2 ml). The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC; Once complete, the reaction mixture was poured into cold water (100 ml) and extracted with ethyl acetate (2x30 ml). The combined organic layers were washed with water (20 ml) and brine (10 ml), then dried over anhydrous Na2SO4. The organic layer was evaporated under reduced pressure to obtain 2.7 g of the desired product. This substance was directly used in the next step without any purification or analysis. The resulting crude compound (2.7 g) was dissolved in toluene (30 ml). To this mixture was added dropwise thionyl chloride (1.78 g, 1.4 eq.) at 0°C and stirred at room temperature for 2 hours. The reaction was monitored by TLC; Once complete, the reaction mixture was poured into ice water (25 ml) and extracted with ethyl acetate (2x25 ml). The combined organic layers were washed with water (30 ml) and brine (10 ml), and dried over anhydrous Na2SO4. The organic layer was evaporated under reduced pressure to obtain 2.4 g of crude compound 7. This material was directly used in the next step without any purification or analysis.

Step 2: synthesis of 1-((4-(4-fluorophenyl)-6-(trifluoromethoxy)-2H-chromen-3-yl)methyl)-4-44 045073 methylpiperazine (I-11).

A mixture of compound 7 (0.2 g, 1 eq.), K ₂ CO ₃ (0.2 g, 3 eq.) and pyrrolidine (0.05 g, 1.2 eq.) in diisopropyl ether (4 ml) was stirred at room temperature for 15 hours. The reaction was monitored by TLC; Once complete, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (2x30 ml). The combined organic layers were washed with water (20 ml) and brine (10 ml), and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography to obtain 22 mg of I-11.

Example 3: 4-((4-(4-fluorophenyl)-6-(trifluoromethoxy)-2H-chromen-3-yl)methyl)morpholine.

Synthesis of 4-((4-(4-fluorophenyl)-6-(trifluoromethoxy)-2H-chromen-3-yl)methyl)morpholine (I-16).

Mixture of compound 7 (0.4 g, 1 eq.), K ₂ CO ₃ (0.5 g, 3 eq.) and morpholine (0.13 g, 1.2 eq.) in diisopropyl ether (4 ml) stirred at room temperature for 15 hours. The reaction was monitored by TLC; Once complete, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (2x30 ml). The combined organic layers were washed with water (20 ml) and brine (10 ml), and then dried over anhydrous sodium sulfate. The crude product was purified by column chromatography to obtain 20 mg of I-16.

Example 4: N-((4-(4-fluorophenyl)-6-(trifluoromethoxy)-2H-chromen-3-yl)methyl)ethanamine.

Synthesis of N-((4-(4-fluorophenyl)-6-(trifluoromethoxy)-2H-chromen-3-yl)methyl)ethanamine (17).

A mixture of compound 7 (0.4 g, 1 eq.), K ₂ CO ₃ (0.5 g, 3 eq.) and ethylamine-HCl (0.12 g, 1.2 eq.) in diisopropyl ether (4 ml ) was stirred at room temperature for 15 hours. The reaction was monitored by TLC; Once complete, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (2x30 ml). The combined organic layer was washed with water (20 ml) and brine (10 ml), and then dried over anhydrous sodium sulfate. The crude product was purified by column chromatography to give 22 mg of 17.

Synthesis scheme 2.

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Example 5: 1-((4-(4-fluorophenyl)-6-(trifluoromethyl)-2H-chromen-3-yl)methyl)pyrrolidine.

F

1-17

Step 1: synthesis of 3-(4-(trifluoromethyl)phenoxy)propanoic acid.

3-Bromopropionic acid (7.0 g, 0.046 mol) was added dropwise over 10 min to a well stirred mixture of compound 8 (5.0 g, 0.033 mol) in DMF (100 ml) at 0°C, then given allowing the temperature to slowly rise to room temperature. Once complete, aqueous NH ₄ Cl (200 ml) was added and the reaction mixture was extracted with EtOAc (2x150 ml). The combined organic layers were washed with H ₂ O (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The organic layer was evaporated under reduced pressure to obtain 2.9 g of crude compound 9, which was directly used in the next step without any purification or analysis.

Step 2: synthesis of 6-(trifluoromethyl)chroman-4-one.

Trifluoromethanesulfonic acid (1.64 ml) was added dropwise over 10 min to a well stirred mixture of compound 9 (2.9 g, 0.012 mmol) in TFA (2.38 ml, 0.03 mol) at 0°C. The reaction mixture was stirred at this temperature for 24 hours. Once complete, the contents were poured into cold H2O (100 ml) and extracted with EtOAc (2x100 ml). The combined organic layers were washed with H ₂ O (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography (hexane:EtOAc; 95:5) to give compound 10 (1-5 g).

Step 3: synthesis of 4-(4-fluorophenyl)-6-(trifluoromethyl)-2H-chromene.

n-BuLi (16.0 mL, 1.6 M in hexane, 6.94 mol) was added dropwise over 10 min to a well-stirred mixture of 3-fluorobromobenzene (1.2 g, 6.94 mol) in THF (20 ml) at -78°C. The reaction temperature was allowed to slowly rise to -50°C and then the mixture was stirred for 1 hour. Compound 10 (1.5 g, 6.94 mol) was dissolved in THF (20 ml) and added dropwise to the reaction mixture at 78° C for 10 minutes and the reaction mixture was allowed to warm to room temperature and then stirred for 2 hours. Once complete, aqueous NH ₄ Cl (200 ml) was added to the reaction mixture, then extracted with EtOAc (2x150 ml). The combined organic layers were washed with H ₂ O (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The organic layer was evaporated under reduced pressure to obtain 2 g of crude product, which was directly used in the next step without any purification or analysis. To this crude product were added an aqueous solution of H ₂ SO ₄ (15 ml, 20%) and 1,4-dioxane (15 ml) and the mixture was refluxed for 2 hours at 105°C. The contents were then washed with aqueous NaOH (20 ml, 1%) and extracted with EtOAc (2x100 ml). The combined organic layers were washed with H ₂ O (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography (hexane:EtOAc; 98:2) to give compound 11 (1.8 g).

Step 4: synthesis of 4-(4-fluorophenyl)-6-(trifluoromethyl)-2H-chromene-3-carbaldehyde.

DMF (1.18 ml, 15.3 mol) and POCl ₃ (1.14 ml, 12.2 mol) were stirred at 0°C for 30 min. Compound 11 (1.8 g, 6.12 mol) in DCM (5 mL) was then added to the well stirred reaction mixture at 0°C. The reaction mixture was refluxed at 45°C for 7 hours. Once complete, the reaction mixture was poured into cold H ₂ O (50 ml) and extracted with EtOAc (2x100 ml). The combined organic layers were washed with saturated sodium bicarbonate (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The organic layer was evaporated under reduced pressure to yield 2 g of crude 12, which was purified by column chromatography (hexane:EtOAc; 98:2) to yield 12 (0.6 g).

Step 5: synthesis of (4-(4-fluorophenyl)-6-(trifluoromethyl)-2H-chromen-3-yl)methanol.

Sodium borohydride (52 mg, 1.39 mol) was added portionwise to a well stirred mixture of compound 12 (0.3 g, 0.9 mol) in methanol (10 ml) at room temperature. The reaction mixture was then stirred at room temperature for 1 hour. The reaction was monitored by TLC; Once complete, the reaction mixture was poured into saturated sodium bicarbonate solution (50 ml) and extracted with ethyl acetate (2x50 ml). The combined organic layers were washed with water (50 ml) and brine (20 ml), separated and then dried over Na ₂ SO ₄ . The organic layer was evaporated under reduced pressure then purified by column chromatography (hexane:EtOAc; 85:15) to give compound 13 (0.3 g).

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Step 6: synthesis of 3-(chloromethyl)-4-(4-fluorophenyl)-6-(trifluoromethyl)-2H-chromene.

Compound 13 (0.3 g) was dissolved in toluene (6 ml) and thionyl chloride (0.1 ml, 1.2 mol) was added dropwise at 0°C. The reaction mixture was then stirred at room temperature for 1 hour and monitored by TLC. Once complete, the mixture was poured into ice water (25 ml) and extracted with EtOAc (2x25 ml). The combined organic layers were washed with water (30 ml) and brine (10 ml), separated and dried over anhydrous Na ₂ SO ₄ . The organic layer was evaporated under reduced pressure to obtain (0.3 g) crude 14, which was directly used in the next step without any purification or analysis.

Step 7: synthesis of 1-((4-(4-fluorophenyl)-6-(trifluoromethyl)-2H-chromen-3-yl)methyl)pyrrolidine (I-17).

A mixture of compound 14 (0.3 g, 0.87 mol), K2CO3 (0.36 g, 2.62 mol) and pyrrolidine (93 mg, 1.31 mol) was stirred in diisopropyl ether (15 ml) at room temperature. The reaction was monitored by TLC; Once complete, the mixture was poured into cold water (20 ml) and extracted with EtOAc (2x30 ml).

The combined organic layers were washed with water (20 ml) and brine (10 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography (hexane:EtOAc; 85:15) to give I-17 (28 mg).

Example 6: (4-(4-fluorophenyl)-6-(trifluoromethyl)-2H-chromen-3-yl)(4-methylpiperazin-1 -yl)methanone.

Step 1: synthesis of 4-phenyl-6-(trifluoromethyl)-2H-chromene-3-carboxylic acid.

Water (0.9 ml) was added dropwise at room temperature to a well stirred mixture of compound 12 (0.3 g, 0.96 mol) Bt-BuOH (10 ml), 2-methyl-2-butene (1.08 g , 9.62 mol), sodium chlorite (0.26 g, 2.88 mol) and NaH ₂ PO ₄ -H ₂ O (0.34 g, 2.88 mol), then stirred for 1.5 hours. The reaction was monitored by TLC; after completion, the mixture was evaporated and the pH of the mixture was adjusted to a value of 2-3 by adding 2N. HCl. The resulting solid precipitated and was isolated by filtration and dried to give compound 15 (0.2 g).

Step 2: synthesis of (4-(4-fluorophenyl)-6-(trifluoromethyl)-2H-chromen-3-yl)(4-methylpiperazin-1yl)methanone (I-26).

Blend of compound 15 (0.2 g, 0.59 mol), EDC (0.17 g, 0.88 mol), N-methylpiperazine (89 mg, 0.88 mol), HOBt (90 mg, 0.59 mol) and DIPEA (0.3 ml, 1.77 mol) were stirred in THF (10 ml) at room temperature. The reaction was monitored by TLC; Once complete, the mixture was poured into ice water and stirred for 30 minutes. The resulting solid was filtered and washed with water. The crude product was purified by column chromatography (DCM:MeOH; 96:4) to give I-26 (22 mg).

Synthesis scheme 3.

Step 1: synthesis of 4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6-ol.

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BBr3 (11.4 g, 1.3 eq.) was added dropwise at 0°C to a solution of 1-((4-(4-fluorophenyl)-6-methoxy-2Hchromen-3-yl)methyl)pyrrolidine 27 (10 g, 1 eq.) in dry DCM (150 ml), then stirred at room temperature overnight. The reaction was monitored by TLC; Once complete, the reaction mixture was poured into ice water and extracted with DCM (2x150 ml). The combined organic layers were washed with water (2x70 ml) and brine (20 ml), and dried over Na ₂ SO ₄ . The crude product was purified by column chromatography (40% ethyl acetate in hexanes) to give 4.7 g M-1.

Step 2: synthesis of 3-((4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6-yl)oxy)propanenitrile (I-32).

K2CO3 (380 mg, 2.7 mmol) was added to a well stirred mixture of compound M-1 (0.3 g, 0.92 mmol) in acrylonitrile (1.5 ml) and DMF (1.5 ml). The reaction mixture was refluxed for 24 hours at 85°C. Once complete, the mixture was poured into cold H2O (200 ml) and subjected to extraction with toluene (2x300 ml). The combined organic layers were washed with H ₂ O (200 ml) and brine (100 ml), separated and then dried over anhydrous Na2SO4. The organic layer was evaporated under reduced pressure to obtain 150 mg of crude I-32, which was directly used in the next step without any further purification.

Example 8: 3-((4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6-yl)oxy)propanoic acid. F ό

1-33

Synthesis of 3-((4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6-yl)oxy)propanoic acid (I-33).

A mixture of compound I-32 (150 mg) in conc. HCl (0.75 ml) was refluxed for 1 hour at 100°C. Once complete, the mixture was poured into cold H ₂ O (200 ml) and the resulting solid was collected by filtration. Purify by column chromatography to give I-33 (100 mg).

Synthesis scheme 4.

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Example 9: 6-fluoro-4-(4-fluorophenyl)-2H-chromen-3-yl)(4-methylpiperazin-1-yl)methanone.

F

1-58

Step 1: synthesis of 3-(4-fluorophenoxy)propanenitrile.

K ₂ CO ₃ (44.4 g, 322 mmol) was added to a well stirred mixture of 4-fluorophenol (compound 17; 12 g, 107.1 mmol) in acrylonitrile (120 ml). The reaction mixture was refluxed for 24 hours at 85°C. Once complete, the mixture was poured into cold H2O (200 ml) and subjected to extraction with toluene (2x300 ml).

The combined organic layers were washed with H2O (200 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The organic layer was evaporated under reduced pressure to obtain 7.2 g of crude compound 18, which was directly used in the next step without any further purification.

Step 2: synthesis of 6-fluorochroman-4-one.

Trifluoromethanesulfonic acid (5.78 ml) was added dropwise over 10 min to a well stirred mixture of compound 18 (7.2 g, 43.6 mmol) in TFA (19.89 g, 174.4 mmol) at 0°C. The reaction mixture was stirred for 24 hours. Once complete, the mixture was poured into cold H ₂ O (100 ml) and extracted with EtOAc (2x150 ml). The combined organic layers were washed with H2O (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography (hexane:EtOAc; 95:5) to give compound 19 (4.6 g).

Step 3: synthesis of 4-(4-fluorophenyl)-6-(trifluoromethoxy)-2H-chromene.

n-BuLi (26 mL, 1.6 M in hexane, 42.0 mmol) was added over 10 min to a well-stirred mixture of 4-fluorobromobenzene (7.3 g, 42.0 mmol) in THF (60 mL) at - 78°C.

The reaction mixture was stirred at -78°C for 30 minutes. Compound 19 (4.6 g, 27.8 mmol) was dissolved in THF (20 ml) and added dropwise over 10 min to the reaction mixture at -78°C, then the reaction temperature was gradually increased to -50°C. The reaction was monitored by TLC; after the starting material was consumed (4 hours), the reaction temperature was raised to -30°C and upon completion, aqueous NH ₄ Cl (50 ml) was added and the reaction mixture was then extracted with EtOAc (2x150 ml). The combined organic layers were washed with H ₂ O (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The organic layer was evaporated under reduced pressure to obtain a crude product which was directly used in the next step without any purification or analysis. To this crude product was added an aqueous solution of H2SO4 (20 ml, 20%) and 1,4-dioxane (50 ml) and the reaction mixture was refluxed for 6 hours at 95°C. The mixture was then washed with aqueous NaOH (20 ml, 1%) and extracted with EtOAc (2x100 ml). The combined organic layers were washed with H ₂ O (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography (hexane) to give compound 20 (3.9 g).

Step 4: synthesis of 6-fluoro-4-(4-fluorophenyl)-2H-chromene-3-carbaldehyde.

DMF (2.92 g, 40.0 mmol) was added dropwise to POCl ₃ (4.90 g, 31.92 mmol) in a round bottom flask, then the mixture was heated to 40°C for 45 min. A solution of compound 20 (3.9 g, 15.98 mmol) in DCM (10 mL) was added dropwise to the same reaction flask and the mixture was heated to 60°C for 15 h. The reaction mixture was cooled to room temperature, diluted with saturated NaHCO ₃ solution (pH ~ 8) and subjected to extraction with EtOAc (2x50 ml). The combined organic layers were washed with H ₂ O (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography (hexane) to give compound 21 (1.5 g).

Step 5: synthesis of 6-fluoro-4-(4-fluorophenyl)-2H-chromene-3-carboxylic acid.

To a well stirred solution of compound 21 (2.6 g, 9.5 mmol) in t-BuOH (52 ml) was added NaH ₂ PO ₄ -2H ₂ O (3.97 g, 28.7 mmol), NaClO ₂ (2 .6 g, 28.7 mmol) and 2-methyl-2-butene (6.72 g, 96.0 mmol) followed by water (2.6 mL) at room temperature, then stirred for 20 min. The reaction was monitored by TLC; upon completion, the mixture was evaporated and the pH was adjusted to a value of 2-3 by adding 2N. HCl. The resulting precipitated solid was isolated by filtration and dried to give compound 22 (1.463 g).

Step 6: synthesis of (6-fluoro-4-(4-fluorophenyl)-2H-chromen-3-yl)(4-methylpiperazin-1-yl)methanone (I-58).

HOBt (159 mg, 1.04 mmol) was added to a mixture of compound 22 (300 mg, 1.04 mmol), Nmethylpiperazine (156 mg, 1.56 mmol), EDC-HCl (299 mg, 1.56 mmol) and DIPEA (400 mg, 3.13 mmol) in

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THF (10 ml) at room temperature. The reaction mixture was then irradiated (microwave radiation) for 3 minutes. Once complete, the reaction mixture was poured into cold H ₂ O (20 ml) and extracted with EtOAc (2x30 ml). The combined organic layers were washed with H ₂ O (20 ml) and brine (10 ml) and separated, and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography to obtain 25 mg of I-58.

Example 10: 1-((6-fluoro-4-(4-fluorophenyl)-2H-chromen-3-yl)methyl)pyrrolidine.

Synthesis of 1 -((6-fluoro-4-(4-fluorophenyl)-2H-chromen-3-yl)methyl)pyrrolidine (I-62).

HOBt (159 mg, 1.04 mmol) was added to a solution of a mixture of compound 22 (300 mg, 1.04 mmol), pyrrolidine (110 mg, 1.56 mmol), EDC-HCl (299 mg, 1.56 mmol) and DIPEA (400 mg, 3.13 mmol) in THF (10 ml) at room temperature. The reaction mixture was then irradiated (microwave radiation) for 3 minutes. Once complete, the reaction mixture was poured into cold H2O (20 ml) and extracted with EtOAc (2x30 ml). The combined organic layers were washed with H2O (20 ml) and brine (10 ml) and separated, and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography to obtain 28 mg of I-62.

Example 11: N ¹ -((6-fluoro-4-(4-fluorophenyl)-2H-chromen-3-yl)methyl)-N ² ,N ² -dimethylethane-1,2-diamine.

Synthesis of N ¹ -((6-fluoro-4-(4-fluorophenyl)-2H-chromen-3-yl)methyl)-N ² ,N ² -dimethylethane-1,2-diamine (I-60).

HOBt (159 mg, 1.04 mmol) was added to a solution of a mixture of compound 22 (300 mg, 1.04 mmol), diamine (140 mg, 1.56 mmol), EDC-HCl (299 mg, 1.56 mmol) and DIPEA (400 mg, 3.13 mmol) in THF (10 ml) at room temperature. The reaction mixture was then irradiated (microwave radiation) for 3 minutes. Once complete, the reaction mixture was poured into cold H2O (20 ml) and extracted with EtOAc (2x30 ml). The combined organic layers were washed with H ₂ O (20 ml) and brine (10 ml) and separated, and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography to obtain 22 mg of I-60.

Example 12: 1-((6-fluoro-4-(4-fluorophenyl)-2H-chromen-3-yl)methyl)-4-methylpiperazine.

Step 1: synthesis of 3-(chloromethyl)-6-fluoro-4-(4-fluorophenyl)-2H-chromene.

To a well stirred mixture of compound 22 (1.5 g, 1 eq.) and sodium borohydride (0.2 g, 1 eq.) in toluene (20 ml), methanol (10 ml) was added dropwise at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored by TLC; Once complete, the reaction mixture was poured into cold water (100 ml) and extracted with ethyl acetate (2x30 ml). The combined organic layer was washed with water (20 ml) and brine (10 ml), and dried over anhydrous sodium sulfate. The organic layer was evaporated under reduced pressure to obtain 0.6 g of the desired product. This substance was directly used in the next step without any purification or analysis. The resulting crude compound (0.6 g) was dissolved in toluene (6 ml) and thionyl chloride (0.344 g, 1.4 eq.) was added dropwise at 0°C. The reaction mixture was then heated and stirred at room temperature for 2 hours. The reaction was monitored by TLC; Once complete, the reaction mixture was poured into ice water (25 ml) and extracted with ethyl acetate (2x25 ml). The combined organic layers were washed with water (30 ml) and brine (10 ml), and dried over anhydrous Na ₂ SO ₄ . The organic layer was evaporated under reduced pressure to obtain 1.5 g of crude compound 23. This material was directly used in the next step without any purification or analysis.

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Step 2: synthesis of 1-((6-fluoro-4-(4-fluorophenyl)-2H-chromen-3-yl)methyl)-4-methylpiperazine (I-59).

A mixture of compound 23 (0.3 g, 1 eq.), K ₂ CO ₃ (0.43 g, 3 eq.), N-methylpiperazine (0.134 g, 1.3 eq.) in diisopropyl ether (10 ml) was stirred at room temperature for 15 hours. The reaction was monitored by TLC; Once complete, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (2x30 ml). The combined organic layers were washed with water (20 ml) and brine (10 ml), and dried over anhydrous Na ₂ SO ₄ . The crude product was purified by preparative TLC to yield 25 mg of I-59.

Example 13: 1-((6-fluoro-4-(4-fluorophenyl)-2H-chromen-3-yl)methyl)pyrrolidine.

Synthesis of 1-((6-fluoro-4-(4-fluorophenyl)-2H-chromen-3-yl)methyl)pyrrolidine (I-63).

A mixture of compound 23 (0.3 g, 1 eq.), K ₂ CO ₃ (0.43 g, 3 eq.), pyrrolidine (0.09 g, 1.2 eq.) in diisopropyl ether (4 ml) was stirred at room temperature for 15 hours. The reaction was monitored by TLC; Once complete, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (2x30 ml). The combined organic layers were washed with water (20 ml) and brine (10 ml), and dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography to obtain 28 mg of I-63.

Example 14: N ¹ -((6-fluoro-4-(4-fluorophenyl)-2H-chromen-3-yl)methyl)-N ² ,N ² -dimethylethane-1,2-diamine.

F

1-61

Synthesis of N ¹ -((6-fluoro-4-(4-fluorophenyl)-2H-chromen-3-yl)methyl)-N ² ,N ² -dimethylethane-1,2-diamine (I-61).

To a mixture of compound 23 (0.25 g, 1 eq.) and ethylamine-HCl (0.08 g, 1.2 eq.) in dry MeOH (5 ml) was added sodium borohydride (0.017 g, 0.5 eq.) and the mixture was stirred at room temperature for 0.5 hours. The reaction was monitored by TLC; Once complete, the reaction mixture was poured into cold water (20 ml) and extracted with ethyl acetate (2x30 ml). The combined organic layers were washed with water (20 ml) and brine (10 ml), and dried over anhydrous Na ₂ SO ₄ . The crude product was purified by preparative TLC to yield 15 mg of I-61.

Synthesis scheme 5.

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Example 15: 4-((4-(4-fluorophenyl)-3-(pyrrolidin-1 -ylmethyl)-2H-chromen-6-yl)oxy)-2-methylbutan-2-ol.

Step 1: synthesis of 4-bromo-2-methylbutan-2-ol.

To a well stirred mixture of ethyl 3-bromopropanoate (compound 24; 2.0 g, 11.04 mmol) in diethyl ether (25 mL), methyl magnesium bromide (11.04 mL, 33.14 mmol) was added dropwise over 10 min at -20°C. The reaction mixture was stirred at room temperature for 2 hours. Once complete, the reaction mixture was poured into aqueous NH ₄ Cl (25 ml) and extracted with EtOAc (2x25 ml). The combined organic layers were washed with H ₂ O (100 ml) and brine (100 ml), separated and then dried over anhydrous Na ₂ SO ₄ . Evaporate under reduced pressure to obtain 2.1 g of crude compound 25, which is directly used in the next step without any further purification.

Step 2: synthesis of 4-((4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6-yl)oxy)-2methylbutan-2-ol (I-35).

Compound 25 (0.24 g, 1.4 mmol) and K ₂ CO ₃ (44.4 g, 322 mmol) were added to a well stirred mixture of compound M1 (0.1 g, 0.3 mmol) in acetonitrile (10 ml ). The mixture was refluxed for 24 hours at 85°C. Once complete, the reaction mixture was poured into cold H ₂ O (200 ml) and extracted with ethyl acetate (2x20 ml). The combined organic layers were washed with H ₂ O (20 ml) and brine (20 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by preparative TLC to give I-35 (20 mg).

Synthesis scheme 6.

Example 16: (4-(4-fluorophenyl)-6-(3-hydroxy-3-methylbutoxy)-2H-chromen-3-yl)(pyrrolidin-1 yl)methanone.

Step 1: synthesis of 4-(4-fluorophenyl)-6-hydroxy-2H-chromene-3-carbaldehyde.

BBr ₃ (2.17 g, 1.3 eq.) was added dropwise to a well stirred solution of 4-(4-fluorophenyl)6-methoxy-2H-chromene-3-carbaldehyde (compound 26; 5 g, 1 eq.) in dry DCM (50 ml) at 0°C. The reaction mixture was then stirred at room temperature overnight. The reaction was monitored by TLC; Once complete, the reaction mixture was poured into ice water and extracted with DCM (2x150 ml). The combined organic layers were washed with water (2x70 ml) and brine (20 ml), and dried over Na ₂ SO ₄ . The crude product was purified by column chromatography (40% ethyl acetate in hexanes) to provide 1.6 g of compound 27.

Step 2: synthesis of 4-(4-fluorophenyl)-6-hydroxy-2H-chromene-3-carboxylic acid.

NaH ₂ PO ₄ -2H ₂ O (2.44 g, 17.7 mmol), NaClO ₂ (1.6 g, 17.7 mmol) and 2-methyl-2-butene (6.25 ml, 59.0 mmol) was added to a well stirred solution of compound 27 (1.6 g, 5.9 mmol) in t-BuOH (15 ml) at room temperature, then water (1.6 ml) was added. The reaction mixture was stirred at room temperature for 20 minutes and monitored by TLC. After completion, the reaction mixture was evaporated and the pH was adjusted to a value of 2-3 by adding 2N. HCl. The resulting precipitated solid was isolated by filtration and dried to give compound 28 (670 mg).

Step 3: synthesis of (4-(4-fluorophenyl)-6-hydroxy-2H-chromen-3-yl)(pyrrolidin-1-yl)methanone.

HOBt (358 mg, 2.34 mmol) was added to a solution of a mixture of compound 28 (670 mg, 2.34 mmol), pyrrolidene (240 mg, 3.56 mmol), EDC-HCl (673 mg, 3.56 mmol) and DIPEA (1.2 mL, 7.0 mmol) in THF (10 mL) at room temperature. The reaction mixture was then irradiated (microwave radiation) for 3 minutes. Once complete, the mixture was poured into cold H2O (20 ml) and extracted with EtOAc (2x30 ml). The combined organic layers were washed with H2O (20 ml) and brine (10 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified to give compound 29 (200 mg).

Step 4: synthesis of (4-(4-fluorophenyl)-6-(3-hydroxy-3-methylbutoxy)-2H-chromen-3-yl)(pyrrolidin-1yl)methanone (I-42).

Compound 25 (0.39 g, 2.3 mmol) and K ₂ CO ₃ (0.1 g, 0.76 mmol) were added to a well stirred mixture of compound 29 (0.2 g, 0.58 mmol) in acetonitrile ( 10 ml). The reaction mixture was refluxed for 24 hours at 85°C. Once complete, the mixture was poured into cold H2O (200 ml) and extracted with ethyl acetate (2x20 ml). The combined organic layers were washed with H2O (20 ml) and brine (20 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by column chromatography to give I-42 (25 mg).

Synthesis scheme 7.

Step 1: synthesis of 3-((4-(4-fluorophenyl)-3-(pyrrolidine-1-carbonyl)-2H-chromen-6-yl)oxy)propanenitrile.

K ₂ CO ₃ (0.24 g, 2.94 mmol) was added to a well stirred mixture of (4-(4-fluorophenyl)-6hydroxy-2H-chromen-3-yl)(pyrrolidin-1-yl)methanone (compound 29 ; 0.2 g, 0.58 mmol) in acrylonitrile (5 ml). The reaction mixture was refluxed for 24 hours at 85°C. Once complete, the mixture was poured into cold H ₂ O (200 ml) and extracted with ethyl acetate (2x25 ml). The combined organic layers were washed with H2O (20 ml) and brine (20 ml), separated and then dried over anhydrous Na2SO4. The crude product was purified by column chromatography (hexane:EtOAc; 50:50) to give compound 30 (0.1 g).

Step 2: synthesis of 3-((4-(4-fluorophenyl)-3-(pyrrolidine-1-carbonyl)-2H-chromen-6-yl)oxy)propanoic acid (I-40).

A mixture of compound 29 (0.1 g) in concentrated HCl solution (0.5 ml) was refluxed for 1 hour at 100°C. Once complete, the mixture was poured into cold H ₂ O (200 ml) and the resulting solid was collected by filtration. Further purification by column chromatography gave I-40 (25 mg).

Synthesis scheme 8.

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Example 18: 1 -(2-((4-(4-fluorophenyl)-3 -(pyrrolidin-1 -ylmethyl)-2H-chromen-6-yl)oxy)ethyl)piperidine.

F

1-39

Step 1: synthesis of 1-((6-(2-bromoethoxy)-4-(4-fluorophenyl)-2H-chromen-3-yl)methyl)pyrrolidine.

Na ₂ CO ₃ (4.78 g, 45.2 mmol) was added to a well stirred mixture of compound M1 (2.45 g, 9.04 mmol) and dibromoethane (0.93 ml, 10.8 mmol) in THF (20 ml).

The reaction mixture was refluxed for 24 hours at 65°C. Once complete, the reaction mixture was poured into cold H2O (200 ml) and extracted with ethyl acetate (2x25 ml). The combined organic layers were washed with H2O (20 ml) and brine (20 ml), separated and then dried over anhydrous Na ₂ SO ₄ .

The crude product was purified by column chromatography (hexane:EtOAc; 40:60) to give compound 31 (0.5 g).

Step 2: synthesis of 1-(2-((4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6yl)oxy)ethyl)piperidine (I-39).

K ₂ CO ₃ (0.75 g, 0.55 mmol) was added to a well stirred mixture of compound 31 (0.16 g, 0.42 mmol) and piperidine (0.16 ml, 1.69 mmol) in acetonitrile (10 ml) The reaction mixture was refluxed for 24 hours at 85°C.

Once complete, the reaction mixture was poured into cold H ₂ O (200 ml) and extracted with ethyl acetate (2x25 ml). The combined organic layers were washed with H ₂ O (20 ml) and brine (20 ml), separated and then dried over anhydrous Na2SO4.

The crude product was purified by preparative TLC to give compound I-39 (18 mg).

Synthesis scheme 9.

Synthesis of 5-(2-((4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6-yl)oxy)ethyl)-1H-tetrazole (I-34).

A catalytic amount of CuI was added to a well stirred mixture of I-32 (0.1 g, 0.26 mmol) and NaN ₃ (0.13 g, 0.52 mmol) in DMF (10 ml). The reaction mixture was heated for 5 hours at 120°C. Once complete, the reaction mixture was poured into cold H ₂ O (200 ml) and extracted with ethyl acetate (2x25 ml). The combined organic layers were washed with H2O (20 ml) and brine (20 ml), separated and then dried over anhydrous Na ₂ SO ₄ . The crude product was purified by preparative HPLC to give compound I-34 (15 mg).

Synthesis scheme 10.

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Example 20: 3-((4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6-yl)oxy)-Nhydroxypropanimidamide.

F

1-36

Synthesis of 3 -((4-(4-fluorophenyl)-3 -(pyrrolidin-1 -ylmethyl)-2H-chromen-6-yl)oxy)-N-hydroxypropanimidamide (I-36).

A solution of I-32 (300 mg, 0.79 mmol), hydroxylamine hydrochloride (110 mg, 1.58 mmol) and sodium carbonate (84 mg, 0.79 mmol) in ethanol (5 ml) and water (1 ml) was heated reflux overnight. The reaction mixture was evaporated under reduced pressure until ethanol was completely removed. This product was collected by filtration and air-dried to give I-36 (100 mg).

Example 21: 3-(2-((4-(4-fluorophenyl)-3-(pyrrolidin-1 -ylmethyl)-2H-chromen-6-yl)oxy)ethyl)-1,2,4oxadiazol-5 (2H )-He.

Synthesis of 3-(2-((4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6-yl)oxy)ethyl)-1,2,4oxadiazol-5(2H)- she (I-37).

A solution of I-36 (100 mg, 0.24 mmol) and carbonyldiimidazole (47 mg, 0.29 mmol) in THF (5 ml) was refluxed for 4 hours. The solvent was removed and the residue was purified by preparative TLC, in resulting in compound I-37 (20 mg).

Example 22: Stability Study in Microsomes.

Stability studies in microsomes were performed using mouse, rat and human cell lines according to techniques well known in the art. The results of the stability study in microsomes are presented in Table. 2 for selected compounds according to the present invention. The connection numbers correspond to the connection numbers in the table. 1. Compounds corresponding to designation A have a half-life of more than thirty (> 30) minutes; compounds designated as B have a half-life of ten (10) to thirty (30) minutes; and compounds designated C have a half-life of less than ten (<10) minutes. N/A means the value is undefined.

table 2

Stability in microsomes

Connection no. Stability in microsomes (Τι/ ₂ min) in mice Stability in microsomes (Τι/ ₂ min) in rats Stability in microsomes (Τι/ ₂ min) in humans 1-1 WITH WITH WITH I-2 WITH WITH IN I-3 WITH WITH IN I-4 IN IN A I-5 V IN IN I-6 A WITH A I-7 WITH WITH IN I-8 WITH WITH A I-9 IN WITH IN 1-10 A A A 1-11 A A A 1-12 A A IN 1-13 WITH WITH IN 1-14 WITH WITH IN 1-15 WITH WITH IN 1-16 IN WITH A

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1-17 A V A 1-18 BUT BUT BUT 1-19 WITH With V I-20 WITH With V 1-21 WITH With V I-22 WITH With V I-23 WITH V V I-24 WITH With With I-25 WITH With V I-26 WITH With V I-27 WITH With With I-28 WITH With With I-29 WITH V With I-30 WITH V A 1-31 WITH With V I-32 IN With V I-33 A A A I-34 A A A I-35 IN With A I-36 IN V A I-37 A V A I-38 BUT BUT BUT I-39 A A A I-40 A A A 1-41 A A WITH I-42 WITH IN WITH I-43 WITH A WITH I-44 WITH WITH WITH I-45 BUT BUT BUT I-46 BUT BUT BUT I-47 BUT BUT BUT I-48 BUT BUT BUT I-49 BUT BUT BUT I-50 BUT BUT BUT 1-51 BUT BUT BUT I-52 BUT BUT BUT I-53 BUT BUT BUT I-54 BUT BUT BUT I-55 BUT BUT BUT I-56 BUT BUT BUT I-57 BUT BUT BUT I-58 With With A I-59 V With A I-60 BUT BUT BUT 1-61 With With IN I-62 BUT BUT BUT I-63 V With A

Example 23: Competitive binding.

Inhibition of the interaction of the SH3 domain of Nck with the proline-rich sequence of the CD3ε TCR subunit was determined by optical readout using a biosensor based on the interferometric optical detection method (IODM) using validated methods (WO 2010/026269; WO 2014/020159; Laguna, M. et al. , Sensors, 2015, 15(8): 19819-29; and Holgado, M. et al., Sensors, 2014, 14(2): 3675-89).

Briefly, a biophotonic cell (BICELL) based on Fabry-Perot interferometers was used as a photonic converter. BICELL was prepared using SU8 in cyclopentanone according to known methods (eg, Laguna, M.F. et al., Sensors, 2014, 14(4): 6695-6700). The cured film was then treated with 95% sulfuric acid for 10 s. Next, streptavidin was immobilized on the sensor surface in phosphate-buffered saline (PBS; pH 7.4) for 20 min at 37°C, then the surface was blocked with 5% skim milk in PBS (pH 7.4). Biotinylated CD3ε peptide (SEQ ID NO: 001) in PBS (pH 7.4) was incubated with streptavidin-coated film to impart biofunctionality to the touch surface (20 min at 37°C).

Competitive binding assays were performed to determine the affinity response of the inhibitors. Screening was performed using a 1:1 ratio between Nck SH3 domain and inhibitors. Optical reading of data from the biosensor was carried out using a Fourier transform spectrometer in the visible and infrared range (FT-VIS-IR). The results of competitive binding for selected compounds according to the present invention are presented in table. 3. The connection numbers correspond to the connection numbers in the table. 1. Compounds meeting the A designation provide an increase in relative optical power (ΔIROP) less than or equal to thirty percent (<30%); compounds designated B provide a ΔIROP of thirty to fifty percent (>30% and <50%); compounds corresponding to designation C provide AIROP from fifty to seventy percent (> 50% and < 70%); and connections corresponding to the designation D provide AIROP

- 56 045073 more than seventy percent (> 70%).

Table 3

Synthesis scheme 11.

MeO

^CN

K ₂ CO3/DMF

1-33

Stage-10

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Example 24: 3-((4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6-yl)oxy)propanoic acid.

1-33

Step 1: synthesis of 3-(4-methoxyphenoxy)propanenitrile.

4-Methoxyphenol (125 g, 1.0 mol) and acrylonitrile (198 ml, 3.0 mol) were placed in a reactor and K ₂ CO ₃ (417 g, 3.0 mol) was added, followed by t-butanol (90 ml) at 25°C. The mass was heated to 80-85°C for 24 hours. The reaction mixture was then cooled to room temperature and filtered to remove inorganic substances. The clear filtrate was quenched with ice water (300 ml) and the aqueous layer was extracted with DCM (300 ml x2). The combined organic layers were dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo to give 3-(4methoxyphenoxy)propanenitrile (160 g, 90%).

Step 2: synthesis of 6-methoxychroman-4-one.

3-(4-Methoxyphenoxy)propanenitrile (160 g, 0.90 mol) and TFA (277 ml, 3.6 mol) were added to the reaction vessel and cooled to 0°C. Trifluoromethanesulfonic acid (120 ml, 1.35 mol) was added slowly through an additional funnel over 20-30 minutes. After addition was complete, the reaction mixture was stirred at 25-30°C for 22-24 hours. The reaction mixture was then quenched with water (480 ml). The resulting mixture was extracted with EtOAc (500 ml x 2). The combined organic layers were washed with brine, dried over anhydrous Na2SO4 and concentrated in vacuo to give a brown gum like 6-methoxy-4-chromanone. Diisopropyl ether (800 ml) was added to the 6-methoxy-4-chromanone resin and stirred for 5-10 minutes. The mixture was cooled to 0°C and stirred for 30 minutes. The reaction mixture was then filtered using a funnel to separate a pale yellow solid. The compound was dried under vacuum at 25-30°C, resulting in 6-methoxychroman-4-one (120 g, 75%).

Step 3: synthesis of 4-(4-fluorophenyl)-6-methoxy-2H-chromene.

The reaction vessel was charged with 1-bromo-4-fluorobenzene (153 ml) and THF (1.125 L) and cooled to 78°C - -70°C. Bu-Li in hexane (1.6 M; 1.0 L) was added dropwise to the reaction mixture at -78°C to -70°C through an additional funnel over 15-20 minutes. 6methoxychroman-4-one (225 g, 1.26 mol) dissolved in THF (1.125 L) was added dropwise to the reaction mixture at -78°C to -70°C through an additional funnel over 15-20 min and reaction the mixture was stirred for 2 hours at the same temperature. After completion of the reaction, a 10% NH ₄ Cl solution was added to the reaction mixture at -78°C -70°C, and the reaction mixture was gradually warmed to room temperature. The reaction mixture was extracted with ethyl acetate (1.125 L) and the combined organic layers were dried over anhydrous Na ₂ SO ₄ . The solvent was distilled off, resulting in a resinous mass of 4-(4-fluorophenyl)-6methoxy chroman-4-ol. The crude alcohol was dissolved in 1,4-dioxane and a 20% H2SO4 solution (1.125 L) was added. The mixture was refluxed and maintained for 1 hour. After completion of the reaction, the reaction mixture was cooled to room temperature and extracted with ethyl acetate (1.125 L). The organic phase was dried over anhydrous Na ₂ SO ₄ and the solvent was completely distilled off, resulting in a brown resinous mass of crude 4-(4-fluorophenyl)-6methoxy-2H-chromene. Methanol (500 ml) was added to the crude 4-(4-fluorophenyl)-6-methoxy-2H-chromene mass and cooled to 0°C. The precipitated solids were filtered to give pure 4-(4-fluorophenyl)-6-methoxy-2H-chromene (160 g, 49%).

Step 4: synthesis of 4-(4-fluorophenyl)-6-methoxy-2H-chromene-3-carbaldehyde.

DMF (75 ml) was added to the reactor and cooled to 0°C. POCl ₃ (73 ml) was slowly added while maintaining the temperature between 0-5°C and stirred for 30 minutes. 4-(4Fluorophenyl)-6-methoxy-2H-chromene (100 g, 0.39 mol) in DCM (250 ml) was added dropwise at 0-10°C. The reaction mixture was allowed to warm to room temperature and then heated to 45°C for 7 hours. Once complete, the reaction mixture was cooled to 20-25°C and then poured into ice water, extracted with ethyl acetate (400 ml x 2), then organic layers dried over anhydrous Na ₂ SO ₄ . The solvent was completely distilled off, resulting in a yellow solid as the crude product. This product was recrystallized in diisopropyl ether to give pure 4-(4-fluorophenyl)-6-methoxy-2H-chromene-3-carbaldehyde (77 g, 69%).

Step 5: synthesis of (4-(4-fluorophenyl)-6-methoxy-2H-chromen-3-yl)methanol.

Methanol (50 ml) was added to a solution of 4-(4-fluorophenyl)-6-methoxy-2H-chromene-3-carbaldehyde (100 g, 0.35 mol) in toluene (400 ml) at 25-30°C. The reaction mixture was cooled to 5-10°C and sodium borohydride was added to the reaction mass in small portions at 5-10°C for 10 minutes. After addition was complete, the reaction mixture was allowed to warm to room temperature and stirred for 1 hour. At the end of this time, 200 ml of water was added to the mixture and the organic layer was washed with brine, dried over anhydrous Na ₂ SO ₄ and concentrated in vacuo. resulting in the title compound (4-(4-fluorophenyl)-6-methoxy-2H-chromen-3yl)methanol (90 g, 90%).

Step 6: synthesis of 3-(chloromethyl)-4-(4-fluorophenyl)-6-methoxy-2H-chromene.

Thionyl chloride (33 ml) was added to a mixture of (4-(4-fluorophenyl)-6-methoxy-2H-chromen-3-yl)methanol (90 g, 0.31 mmol) in toluene (200 ml) at 5-10° C and then stirred at 5-10°C for 1 hour. The reaction mixture was quenched with water (2500 ml) and stirred for 15 minutes, and the organic layer was dried over anhydrous Na2SO4 and concentrated in vacuo to give 3-(chloromethyl )-4-(4fluorophenyl)-6-methoxy-2H-chromene in the form of a light brown resinous mass (83 g, 88%).

Step 7: synthesis of 1-((4-(4-fluorophenyl)-6-methoxy-2H-chromen-3-yl)methyl)pyrrolidine.

Pyrrolidine (33 ml) was added to a solution of 3-(chloromethyl)-4-(4-fluorophenyl)-6-methoxy-2H-chromene (100 g, 0.33 mmol) in diisopropyl ether (400 ml) and the mixture was cooled to 5 -10°C. Anhydrous potassium carbonate (138 g, 1.0 mol) was added at 5-10°C and the reaction mixture was then heated to 55-60°C for 1 hour. After this time, the reaction mass was cooled to 25-30°C and filtered . The clear filtrate was washed with water (250 ml) and the organic layer was dried over anhydrous sodium sulfate and concentrated to obtain a light brown gummy mass of 1-((4(4-fluorophenyl)-6-methoxy-2H-chromene-3- yl)methyl)pyrrolidine (100 g, 89%).

Step 8: synthesis of 1-((4-(4-fluorophenyl)-6-methoxy-2H-chromen-3-yl)methyl)pyrrolidine hydrochloride.

HCl in diisopropyl ether (250 ml) was added dropwise to a solution of 1-((4-(4-fluorophenyl)-6methoxy-2H-chromen-3-yl)methyl)pyrrolidine (100 g, 0.29 mol) in diisopropyl ether (400 ml) between 5-10°C for 1 hour, then stirred for 30 minutes at 5-10°C. The precipitated solid was filtered and dried under vacuum at 40-45°C to give 1-((4-(4-fluorophenyl)-6methoxy-2H-chromen-3-yl)methyl)pyrrolidine hydrochloride (100 g, 91 %).

Step 9: synthesis of 4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6-ol.

Pure BBr ₃ (100 g, 38 ml, 0.4 mol) was added to a solution of 1-((4-(4-fluorophenyl)-6-methoxy-2Hchromen-3-yl)methyl)pyrrolidine hydrochloride (100 g, 0. 27 mol) in dry DCM (1500 ml) at 0-10°C and after addition was complete, the mixture was stirred at room temperature for 16 hours. The DCM layer was decanted, separating the resinous mass. This product was quenched with methanol and diluted with water to precipitate the product.

The precipitated solid was filtered and dried by drainage to give 4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6-ol (75 g, 86%).

Step 10: synthesis of 3-((4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6-yl)oxy)propanenitrile.

DMF (125 ml) and potassium carbonate (25.5 g, 185.0 mmol) were added to a solution of 4-(4-fluorophenyl)-3(pyrrolidin-1-ylmethyl)-2H-chromen-6-ol (25 g, 61.7 mmol) in acrylonitrile (250 ml). The mixture was heated to 80-85°C for 24 hours. After this time, the reaction mixture was concentrated under vacuum. The resulting residue was quenched with water and extracted with ethyl acetate (300 ml). The organic layer was dried over anhydrous sodium sulfate and concentrated to give a light brown gummy mass, which was further purified by column chromatography to give 3-((4-(4-fluorophenyl)-3-(pyrrolidine- 1-ylmethyl)-2H-chromen-6yl)oxy)propanenitrile as a light brown solid (11 g, 47%).

Step 11: synthesis of 3-((4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2H-chromen-6yl)oxy)propanoic acid hydrochloride.

Concentrated HCl (1.0 L) was added to 3-((4-(4-fluorophenyl)-3-(pyrrolidin-1-ylmethyl)-2Hchromen-6-yl)oxy)propanenitrile (100 g, 26 mol) and the reaction the mixture was heated to 95-100°C for 4 hours. The reaction mixture was then cooled to room temperature and stirred for 3 hours. The precipitated solids were filtered and dried by drainage to obtain crude I-33 (60 g) with HPLC purity 92-95% a/a.

Example 25: CD3-mediated phosphorylation of ZAP70 in Jurkat cells.

Inhibition of CD3-mediated phosphorylation of ZAP70 in Jurkat cells was determined by cell-specific homogeneous time-resolved fluorescence (HTRF) assay using a total ZAP-70 HTRF assay kit. The materials used for the HTRF analysis are given in Table. 4 below.

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Table 4

Materials for pZAP70 inhibition

Materials Catalog number Provider Cell culture medium RPMI 1640 R6504 Sigma-Aldrich PBS 10010031 Thermo Fisher Scientific B.S.A. B6917 Sigma-Aldrich Microplates with fine wells (white) ProxiPlate-384 Plus 6008281 PerkinElmer Anti-CD3 monoclonal antibody (OCT3) functional class 16-0037-81 Thermo Fisher Scientific Histopaque-1077 10771 Sigma-Aldrich Total ZAP-70 Determination Kit Using HTRF 64ZATPEG Cisbio HBSS 14025092 Thermo fisher

Briefly, Jurkat cells (~1.3 x 10 ⁶ cells/ml) were collected on the day of the experiment. Cells were washed with PBS, resuspended in assay buffer (RPMI 1640 medium plus 10% fetal bovine serum), and counted.

Cell counts were performed by diluting 10 μl of cells with 90 μl of medium/PBS (1:10). Twenty (20) µl of the diluted cell suspension was then added to 20 µl of trypan blue solution (1:1) and mixed thoroughly to avoid aerosol formation. The cell culture mixture was then placed in a hemocytometer until the cover slip was sufficiently filled and the suspension was allowed to settle in the hemocytometer for at least 10 min before being counted. Viable cells (colorless) and dead cells (blue) were counted in four (4) 1 mm squares at the corners, including cells touching the top line or the vertical perimeter line of each corner area, but not including cells touching either the bottom line or the right line vertical perimeter of the area of any angle.

The number of viable cells per ml was calculated as follows.

Cells/ml = Cells in all four squares x 10 x 2 x 10 ⁴ /4

10 ⁴ = conversion factor for volume of 1 ml; 10 = cell suspension dilution factor; 2 = trypan blue dilution factor

Total cell count = number of cells/ml x total volume (ml) of cell suspension % cell viability = (Counted number of viable cells/total number of counted cells (viable + dead)) x 100

Jurkat cells were seeded into assay medium in a total volume of 4 μl/well (40K cells per well) in a 384-well flat-bottomed fine-well plate. Cells were then incubated at 37°C/5% CO ₂ for 30 min. Next, the cells were treated in duplicate with test compounds (3x) and reference compounds (3x) in a total volume of 4 μl, respectively. The dilution scheme for the compound is illustrated in Table. 5 below.

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Table 5

Illustrative scheme for diluting compounds

100% DMSO Ref. conc.(mM) 20 mM — 2 µl (20mM) + 98.25 µl (100% DMSO) — f X*”^. X X* x^^\ x^^\ x* 3.125-fold dilution (16 µl compound + 34 µl DMSO 100% DMSO Razb. ref. solution (µM) 399 133.0000 44.3333 14.7778 4.9259 1.6420 0.5473 0.1824 0.0608 0.0203 0.0068 1 µl compound + 132 µl RPMI medium 1 1 1 0.75% DMSO Intermediate disag. 3.0000 1.0000 0.3333 0.1111 0.0370 0.0123 0.0041 0.0014 0.0005 0.0002 0.0001 4 µl of compound in a total reaction volume of 12 µl X X 7 X 0.25% DMSO Final concentration (µM) 1.000000 0.333333 0.111111 0.037037 0.012346 0.004115 0.001372 0.000457 0.000152 0.000051 0.000017

The plate was then transferred to a 37°C/5% CO2 incubator for 2 min. After incubation, cells were stimulated with 20 μg/ml anti-CD3 antibody (1 mg/ml stock) and assay media was used to bring the total volume to 4 μl per well. The plate was then transferred to a 37°C/5% CO ₂ incubator for 2.5 min. After incubation, 4 μl of the lysis buffer provided in the kit (4x) was added to each well. Next, the contents of the plate were incubated at room temperature for 1 hour with gentle shaking. After incubation, 4 μl of detection mixture was added to each well, the plate was tightly sealed using a plate sealer, and left overnight at room temperature in the dark. An Envision plate reader was set to Eu ³⁺ cryptate and fluorescence emission was determined at two different wavelengths (620 nm and 665 nm).

Excitation filter: UV2(TRF) 320 nm.

Emission filter: 665 nm.

^2nd emission filter: europium, 615 nm.

Definition height (mm): 6.5.

Exciting light (%): 100.

Latency (μs): 50.

Time window (μs): 400.

Time between flashes (µs): 2000.

Number of flashes: 100.

Number of flashes for the 2nd detector: 100.

The data is presented as a signal at two wavelengths:

Signal _bb5 im

₆₂₀ nm signal

The signal ratio is calculated for each individual well:

Standard deviation ---------------------x 100

Average ratio

Data were plotted using Graph Pad Prism. The significance of the differences between the untreated group and the compound treated groups was assessed using one-way analysis of variance and a Bonferroni post hoc test comparing all columns in the graph.

The results of determining IC ₅₀ in relation to ZAP70 are presented in table. 6 for selected compounds according to the present invention. The connection numbers correspond to the connection numbers in the table. 1. Compounds designated A have an IC ₅₀ of less than one nanomol/L (< 1 nM). Compounds designated B have an IC ₅₀ of one and less than ten nanomol/L (1 nM < x < 10 nM). Compounds corresponding to the designation C have an IC ₅₀ of ten or less than fifty nanomol/l (10 nM < x < 50 nM). Compounds designated D have an IC ₅₀ greater than fifty nanomol/L (>50 nM). BUT means not defined.

The results for determining the maximum percentage of ZAP70 inhibition are presented in table. 7 for selected compounds according to the present invention. The connection numbers correspond to the connection numbers in the table. 1. Compounds meeting the designation A provide a maximum percentage of inhibition greater than fifty percent (>50%). Compounds corresponding to the designation B provide a maximum percentage of inhibition of up to fifty percent or more than forty percent (50% > x > 40%). Compounds corresponding to the designation C provide a maximum percentage of inhibition of up to forty percent or more than thirty percent (40% > x > 30%). Compounds meeting the designation D provide a maximum percentage of inhibition of thirty percent or less (<30%).

Claims (20)

1. A compound selected from any of the compounds of formulas IV-a, IV-b, IV-c or IV-d: IV-c IV-d or a pharmaceutically acceptable salt thereof, wherein R ¹ is R, halogen, -CN, -OR, -NHR or -N(R)2; each R independently represents hydrogen, a C _1-6 straight or branched saturated or unsaturated hydrocarbon chain, a 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring, phenyl, a 4-8 membered saturated or partially unsaturated monocyclic heterocyclic ring containing 1 -2 heteroatoms independently selected from nitrogen, oxygen or sulfur, a 5-6 membered monocyclic heteroaromatic ring containing 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, where each substituent is from the specified hydrocarbon chain, carbocyclic ring, phenyl, heterocyclic ring and heteroaromatic ring optionally substituted 1-6 times by deuterium, halogen, -OH, -CN, -COOH, -C(=N-OH)NH ₂ , C _1-3 straight or branched saturated or unsaturated hydrocarbon chain, optional 1-3 times substituted by deuterium, halogen or -OH, a 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring or phenyl, or two R at the same nitrogen atom are combined with the atoms located between them to form a 5-6 membered heterocyclic a ring containing 0-1 heteroatom in addition to a nitrogen atom attached to said ring, wherein said heteroatom is oxygen, nitrogen or sulfur, wherein said heterocyclic ring is optionally substituted 1-6 times by deuterium, halogen, -OH, -CN, - COOH, -C(=N-OH)NH ₂ , C _1-3 straight or branched saturated or unsaturated hydrocarbon chain, optionally 1-3 times substituted by deuterium, halogen or -OH, 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring or phenyl; L ² represents a covalent bond or a C _1-4 bivalent straight or branched saturated or unsaturated hydrocarbon chain in which 1-2 methylene chain units are independently and optionally replaced by -O-, -C(O)-, - C(O)O-, -OC(O)-, -C(S)-, -C(R)2-, -CH(R)-, -C(F) ₂ -, -N(R)-, -C(O)N(R)-, -N(R)C(O)-, -OC(O)N(R)- or -N(R)C(O)N (R)-; and wherein said compound is not a compound selected from: F F F 12 3 F F F 2. A compound according to claim 1, selected from any of the compounds of formulas V-a, VI-a, V-c or VI-c: - 64 045073 R ³ or a pharmaceutically acceptable salt thereof. 3. Compound of formula IX: IX or a pharmaceutically acceptable salt thereof, where R ² is R, halogen, -C(O)N(R) ₂ or -N(R) ₂ ; R ³ is hydrogen, halogen, -CN or -NO ₂ ; each R independently represents hydrogen, a C _1-6 straight or branched saturated or unsaturated hydrocarbon chain, a 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring, phenyl, a 4-8 membered saturated or partially unsaturated monocyclic heterocyclic ring containing 1 -2 heteroatoms independently selected from nitrogen, oxygen or sulfur, a 5-6 membered monocyclic heteroaromatic ring containing 1-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, where each substituent is from the specified hydrocarbon chain, carbocyclic ring, phenyl, heterocyclic ring and heteroaromatic ring optionally substituted 1-6 times by deuterium, halogen, -OH, -CN, -COOH, -C(=N-OH)NH ₂ , C _1-3 straight or branched saturated or unsaturated hydrocarbon chain, optional 1-3 times substituted by deuterium, halogen or -OH, a 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring or phenyl, or two R at the same nitrogen atom are combined with the atoms located between them to form a 5-6 membered heterocyclic a ring containing 0-1 heteroatom in addition to a nitrogen atom attached to said ring, wherein said heteroatom is oxygen, nitrogen or sulfur, wherein said heterocyclic ring is optionally substituted 1-6 times by deuterium, halogen, -OH, -CN, - COOH, -C(=N-OH)NH ₂ , C _1-3 straight or branched saturated or unsaturated hydrocarbon chain, optionally 1-3 times substituted by deuterium, halogen or -OH, 3-8 membered saturated or partially unsaturated monocyclic carbocyclic ring or phenyl; L ² represents a covalent bond or C _1-4 bivalent straight or branched saturated or unsaturated hydrocarbon chain in which 1-2 methylene chain units are independently and optionally replaced by -O-, -C(O)-, -C(O) O-, -OC(O)-, -C(S)-, -C(R) ₂ -, -CH(R)-, -C(F) ₂ -, -N(R)-, -C( O)N(R)-, -N(R)C(O)-, -OC(O)N(R)- or -N(R)C(O)N(R)-. 4. The compound according to claim 3 or a pharmaceutically acceptable salt thereof, where R ² represents -N(R) ₂ , where two R at the nitrogen atom are combined with the atoms located between them to form a 5-6 membered heterocyclic ring containing no heteroatom in addition to a nitrogen atom attached to said ring, and wherein said 5-6 membered heterocyclic ring is optionally substituted 1-6 times by a C _1-3 straight or branched saturated or unsaturated hydrocarbon chain or halogen. 5. The compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein R ³ represents fluorine. 6. A compound according to claim 3 or a pharmaceutically acceptable salt thereof, wherein L ² represents a C _1-4 bivalent straight or branched saturated or unsaturated hydrocarbon chain, in which 1 methylene chain unit is optionally replaced by -C(O)-. 7. Compound of formula IX: - 65 045073 or a pharmaceutically acceptable salt thereof, wherein R ² represents -N(R) ₂ , where two R at the same nitrogen atom are combined with the atoms located between them to form a 5-6-membered heterocyclic ring that does not contain a heteroatom in addition to the nitrogen atom attached to the specified ring, and wherein said 56-membered heterocyclic ring is optionally substituted 1-6 times by a C _1-3 straight or branched saturated or unsaturated hydrocarbon chain or halogen; R ³ represents halogen; L ² represents a C _1-4 bivalent straight or branched saturated or unsaturated hydrocarbon chain in which 1 methylene unit of the chain is optionally replaced by -C(O)-. 8. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, where R ² represents 9. The compound according to claim 8 or a pharmaceutically acceptable salt thereof, where R ² represents ⁴ - 10. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein R ³ represents fluorine. 11. A compound according to claim 7 or a pharmaceutically acceptable salt thereof, wherein L ² represents -C(O)-. 12. The compound according to claim 7 or a pharmaceutically acceptable salt thereof, where L ² represents -CH ₂ or -(CH ₂ ) ₂ -. 13. A connection selected from the group consisting of: - 66 045073 - 67 045073 - 68 045073 - 69 045073 - 70 045073 - 71 045073 - 72 045073 or a pharmaceutically acceptable salt thereof. 14. A compound of formula or a pharmaceutically acceptable salt thereof. 15. A pharmaceutically acceptable composition comprising a compound according to any one of claims 1 to 14 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient, adjuvant or carrier. 16. A method of treating a TCR-Nck mediated disease, disorder or condition in a patient in need thereof, comprising administering to said patient a compound of any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable composition thereof of claim 15. 17. A method of treating a disease, disorder or condition in a patient in need thereof, comprising administering to said patient a compound of any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable composition thereof of claim 15, wherein said disease, the disorder or condition is selected from rheumatoid arthritis, psoriatic arthritis, psoriasis, type I diabetes, complications of diabetes, multiple sclerosis, systemic lupus erythematosus, cutaneous lupus erythematosus, atopic dermatitis, mast cell-mediated allergic reactions, autoimmune hepatitis, myasthenia gravis, ankylosing spondylitis, Crohn's disease, leukemia, lymphomas and thromboembolic and allergic complications associated with leukemia and lymphomas. 18. The method of claim 17, wherein the disease, disorder or condition is leukemia or lymphoma. 19. The method according to claim 17, where the leukemia is selected from acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia, acute promyelocytic leukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acute erythroid leukemia, chronic leukemia, chronic myelocytic leukemia for, chronic lymphocytic leukemia, B-cell prolymphocytic leukemia, Burkitt leukemia and mast cell leukemia. 20. The method according to claim 17, where the lymphoma is selected from diffuse large B-cell lymphoma, - 73 045073 follicular lymphoma, chronic lymphocytic lymphoma, lymphoplasmacytic lymphoma, marginal zone lymphoma of the spleen, non-Hodgkin lymphoma, Hodgkin lymphoma, extranodal B-cell marginal zone lymphoma, nodal B-cell marginal zone lymphoma, mantle cell lymphoma, large B-cell lymphoma of the mediastinum ( thymus), intravascular large B-cell lymphoma, primary effusion lymphoma, Burkitt's lymphoma, or lymphomatoid granulomatosis.