Novel lymphoid-specific tyrosine phosphatase (LYP) inhibitors

Abstract

The present disclosure relates to novel lymphoid-specific tyrosine phosphatase (LYP (encoded by the PTPN22 gene)) inhibitors, as well as methods for making and using the novel LYP inhibitors. Thus, compounds according to the present disclosure can be used to treat diseases or disorders associated with PTPN22 gene polymorphisms, including type 1 diabetes, rheumatoid arthritis, systemic lupus erythematosus, Graves' disease, Addison's disease, vitiligo, juvenile arthritis, Hashimoto's thyroiditis, and other additional rare diseases. Furthermore, these LYP inhibitors may be useful in a novel class of cancer immunotherapy. The compounds can be injected or administered orally.

Description

[Technical Field]

[0001] Government Rights This invention was made with government support under Grant No. RO1 CA207288 awarded by the National Institutes of Health (NIH). The government has certain rights in this invention.

[0002] Technical Field The present disclosure relates to novel lymphoid-specific tyrosine phosphatase (LYP) inhibitors and methods of making and using the novel lymphoid-specific tyrosine phosphatase (LYP) inhibitors. [Background technology]

[0003] background This section introduces aspects that may be helpful in facilitating a better understanding of the present disclosure. Accordingly, these statements are to be read and understood in this light, and not as admissions about what is prior art or what is not.

[0004] Lymphoid-specific tyrosine phosphatase (LYP) is a non-receptor protein tyrosine phosphatase (PTP) (encoded by the PTPN22 gene). It is a 110-kDa protein consisting of an N-terminal PTP domain and a non-catalytic C-terminal segment with several Pro-rich motifs. Human genetic studies have shown that single-nucleotide polymorphisms in PTPN22 are frequently mutated in patients with autoimmune diseases (e.g., type 1 diabetes, rheumatoid arthritis, systemic lupus erythematosus, Graves' disease, Addison's disease, vitiligo, juvenile arthritis, and Hashimoto's thyroiditis). Biochemical studies suggest that LYP inhibits T cell activation, possibly through dephosphorylation of T cell receptor (TCR)-associated Lck and ZAP-70 kinases. Given its important role in regulating TCR signaling pathways, LYP has recently emerged as a potential target for the treatment of a wide range of autoimmune diseases.

[0005] LYP has also been found to be a critical negative regulator of antitumor T cell responses. Adoptive cell therapy studies have shown that CD8+ T cells lacking PTPN22 are superior at eliminating established tumors. A major impact of PTPN22 deficiency is that it confers an enhanced ability to produce proinflammatory cytokines and kill tumors expressing low-affinity tumor-associated antigens to both effector and memory CD8+ T cells. Furthermore, the absence of PTPN22 promotes macrophage polarization toward a proinflammatory M1 phenotype and increases CD40 expression on dendritic cells (resulting in higher proliferation of cocultured CD4+ T cells). This evidence provides a strong rationale for targeting LYP in T cells for cancer immunotherapy.

[0006] Therefore, given the strong association of LYP with autoimmunity, small molecule LYP inhibitors may have therapeutic value for treating diseases or disorders associated with PTPN22 gene polymorphisms. Furthermore, the successful development of small molecule inhibitors of LYP would also provide a new class of cancer immunotherapeutic agents and facilitate the development of novel combination strategies with LYP inhibitors.

[0007] However, the design and synthesis of inhibitors of LYP with optimal potency, selectivity, and pharmacological properties remains a challenging endeavor. Summary of the Invention [Means for solving the problem]

[0008] Abstract The present disclosure relates to novel lymphoid-specific tyrosine phosphatase (LYP) inhibitors and methods of making and using the novel lymphoid-specific tyrosine phosphatase (LYP) inhibitors.

[0009] In one embodiment, the present disclosure provides a compound of formula I: [ka] or a stereoisomer, tautomer, solvate, derivative, or pharmaceutically acceptable salt thereof, wherein: R 1 and R 2 independently represent hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, alkylcarbonyl, provided that said alkylcarbonyl is not methylcarbonyl, optionally substituted cycloalkylcarbonyl, optionally substituted cycloalkylalkylcarbonyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted arylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted alkoxyalkyl, optionally substituted hydroxyalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylcarbonyl, optionally substituted heterocyclylalkylcarbonyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted amino, optionally substituted alkyl-SO2-, optionally substituted aryl-SO2-, optionally substituted heterocyclyl-SO2-, optionally substituted amino-SO2-, or R 1 and R 2 together with the N atom to which they are attached form a 5-10 membered heterocyclic ring optionally containing a second heteroatom selected from nitrogen or oxygen, wherein said heterocyclic ring is optionally substituted; R 3 is H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heterocyclyl; R 4 and R 6represent independently hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and heteroaryl; optionally substituted aralkyl, or optionally substituted heteroaralkyl; (R 5 ) n when n is 1 or 2, represents 1 to 2 of -H, -F, -Cl, -Br, -I, -CFH2, -CF2H, -CF3, -CN, -OH, -NO2, -NH2, -SO2CH3, SO2NH2, -SON2NHCH3, optionally substituted -CO2-alkyl, optionally substituted NH(alkyl) or N(alkyl)2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkoxy, optionally substituted S-alkyl, optionally substituted cycloalkyl, optionally substituted heterocycle, optionally substituted aryl or optionally substituted heteroaryl; X is O or H, where -C=X- is -CH2- when X is H, where R 1 and R 2 is optionally substituted arylcarbonyl when X is H, and R 1 and R 2 One of them is R 3 to form a ring; Y is CR 5 Or N. BRIEF DESCRIPTION OF THE DRAWINGS [Brief explanation of the drawings]

[0010] [Figure 1] Figure 1 illustrates the selectivity data of Example 1 for LYP and its inhibitory activity against a panel of 16 mammalian PTPs. Selectivity profiling revealed that Example 1 exhibits 7-10 fold greater selectivity for LYP over similar phosphatases.

[0011] [Figure 2] FIG. 2 illustrates the kinetic analysis showing Example 1 as a competitive inhibitor of LYP with Ki=0.50±0.03 μM.

[0012] [Figure 3] Figures 3 and 4 illustrate in vivo pharmacokinetic data based on mass spectrometry quantification at 0 hours, 1.0 hours, 1.5 hours, 2.0 hours, 2.5 hours, 3.0 hours, 6.0 hours, and 24.0 hours from the time of intraperitoneal injection of Example 1 for three mice. [Figure 4] Figures 3 and 4 illustrate in vivo pharmacokinetic data based on mass spectrometry quantification at 0 hours, 1.0 hours, 1.5 hours, 2.0 hours, 2.5 hours, 3.0 hours, 6.0 hours, and 24.0 hours from the time of intraperitoneal injection of Example 1 for three mice.

[0013] [Figure 5] FIG. 5 illustrates that Example 1 demonstrates effective in vivo activity in a mouse model for cancer immunotherapy. DETAILED DESCRIPTION OF THE INVENTION

[0014] Detailed Description For the purposes of promoting an understanding of the principles of the present disclosure, reference will now be made to the embodiments illustrated in the drawings and specific language will be used to describe the same, nevertheless it will be understood that no limitation of the scope of this disclosure is thereby intended.

[0015] In this disclosure, the term "about" may allow for a degree of variability in a value or range (e.g., within 10%, within 5%, or within 1% of a stated value or a stated boundary of a range).

[0016] In this disclosure, the term "substantially" may allow for a degree of variability in a value or range (e.g., within 90%, within 95%, or within 99% of a stated value or of a stated boundary of a range).

[0017] The term "substituted," as used herein, refers to a functional group in which one or more hydrogen atoms contained therein are replaced with one or more non-hydrogen atoms. The term "functional group" or "substituent," as used herein, refers to a group that can be present on a molecule or can be substituted on a molecule. Examples of substituents or functional groups include, but are not limited to, halogens (e.g., F, Cl, Br, and I); oxygen atoms in groups such as hydroxyl groups, alkoxy groups, aryloxy groups, aralkyloxy groups, oxo(carbonyl) groups, carboxyl groups (including carboxylic acids, carboxylates, and carboxylic acid esters); sulfur atoms in groups such as thiol groups, alkyl and aryl sulfide groups, sulfoxide groups, sulfone groups, sulfonyl groups, and sulfonamide groups; nitrogen atoms in groups such as amines, azides, hydroxylamines, cyanos, nitro groups, N-oxides, hydrazides, and enamines; and other heteroatoms in various other groups.

[0018] Non-limiting examples of substituents that may be attached to a substituted carbon (or other, such as nitrogen) atom include: F, Cl, Br, I, OR, OC(O)N(R), CN, NO, NO, ONO, azido, CF, OCF, R, O(oxo), S(thiono), C(O), S(O), methylenedioxy, ethylenedioxy, N(R), SR, SOR, SO, R, SO, N(R), SO, R, (CH). 0-2 P(O)OR2, C(O)R, C(O)C(O)R, C(O)CH2C(O)R, C(S)R, C(O)OR, OC(O)R, C(O)N(R)2, OC(O)N(R)2, C(S)N(R)2, (CH2) 0-2 N(R)C(O)R, (CH2) 0-2 N(R)C(O)OR, (CH2) 0-2N(R)N(R), N(R)N(R)C(O)R, N(R)N(R)C(O)OR, N(R)N(R)CON(R), N(R)SOR, N(R)SON(R), N(R)C(O)OR, N(R)C(O)R, N(R)C(S)R, N(R)C(O)N(R), N(R)C(S)N(R), N(COR)COR, N(OR)R, C(=NH)N(R), C(O)N(OR)R, or C(=NOR)R, where R is hydrogen or a carbon-based moiety, which may itself be further substituted; e.g., where R is , hydrogen, alkyl, acyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl, or heteroarylalkyl, where any alkyl, acyl, cycloalkyl, aryl, aralkyl, heterocyclyl, heteroaryl, or heteroarylalkyl or R can be independently mono- or polysubstituted; or where two R groups are attached to one nitrogen atom or to adjacent nitrogen atoms to form, together with the nitrogen atom(s), a heterocyclyl that can be mono- or independently polysubstituted.

[0019] The term "alkyl", alone or in combination, denotes a straight-chain or branched alkyl group having 1 to 8 carbon atoms, preferably a straight-chain or branched alkyl group having 1 to 6 carbon atoms, and particularly preferably a straight-chain or branched alkyl group having 1 to 4 carbon atoms. Examples of straight-chain or branched C1-C8 alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, the isomeric pentyls, isomeric hexyls, isomeric heptyls, and isomeric octyls, preferably methyl and ethyl, and most preferably methyl.

[0020] The term "cycloalkyl," alone or in combination, refers to a cycloalkyl ring having 3 to 8 carbon atoms and preferably a cycloalkyl ring having 3 to 6 carbon atoms. Examples of C3-C8 cycloalkyl are cyclopropyl, methyl-cyclopropyl, dimethyl-cyclopropyl, cyclobutyl, methyl-cyclobutyl, cyclopentyl, methyl-cyclopentyl, cyclohexyl, methyl-cyclohexyl, dimethyl-cyclohexyl, cycloheptyl and cyclooctyl, preferably cyclopropyl and especially cyclopentyl.

[0021] The term "alkoxyl", alone or in combination, denotes a radical of the formula alkyl-O-, wherein the term "alkyl" has the meaning given above (e.g., methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, isobutoxy, sec-butoxy and tert-butoxy, 2-hydroxyethoxy, 2-methoxyethoxy, preferably methoxy and ethoxy, and most preferably methoxy).

[0022] The term "aryl," as used herein, refers to a substituted or unsubstituted cyclic aromatic hydrocarbon that does not contain heteroatoms in the ring. Thus, aryl groups include, but are not limited to, phenyl, azulenyl, heptalenyl, biphenyl, indacenyl, fluorenyl, phenanthrenyl, triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenylenyl, anthracenyl, and naphthyl groups. In some embodiments, an aryl group contains from about 6 to about 14 carbons (C6-C8) in the ring portion of the group. 14 ) or 6 to 10 carbon atoms (C6-C 10) Aryl groups can be unsubstituted or substituted as defined herein. The term "aryl," alone or in combination, further refers to a phenyl or naphthyl group, preferably a phenyl group optionally bearing one or more, especially one to three, substituents, each independently selected from halogen, trifluoromethyl, amino, alkyl, alkoxy, aryloxy, alkylcarbonyl, cyano, carbamoyl, alkoxycarbamoyl, methylenedioxy, carboxy, alkoxycarbonyl, aminocarbonyl, alkylaminocarbonyl, dialkylaminocarbonyl, hydroxyl, nitrol, and the like. Preferred substituents for aryl, preferably phenyl, are independently selected from halogen, trifluoromethyl, alkyl, alkoxy, cyano, and nitro. Examples of aryl are phenyl, cyanophenyl, methoxyphenyl, fluorophenyl, and methylphenyl.

[0023] The term "aralkyl", alone or in combination, denotes an alkyl or cycloalkyl group as defined above, substituted with one or more, preferably one or two, particularly preferably one, aryl group, and wherein the term aryl is defined as above. Examples are benzyl, benzyl substituted with hydroxyl, alkoxy or halogen, preferably fluorine.

[0024] The term "heterocyclyl," alone or in combination, refers to a saturated, partially unsaturated, or aromatic 4- to 10-membered heterocycle containing one or more, preferably one or two, heteroatoms selected from nitrogen, oxygen, and sulfur, where oxygen and especially nitrogen are preferred. If desired, it may be substituted at one or more carbon atoms by halogen, alkyl, alkoxy, oxo, alloxyalkyl, hydroxyalkyl, etc., and / or on the secondary nitrogen atom (i.e., -NH-) by alkyl, cycloalkyl, arakoxycarbonyl, alkanoyl, phenyl, or phenylalkyl, or on the tertiary nitrogen atom (i.e., =N-) by oxide (halogen, alkyl, cycloalkyl, and alkoxy are preferred). Examples of such heterocyclyl groups are pyridinyl, furyl, pyrrolidinyl, piperidinyl, morpholinyl, piperazinyl, 3,4-dihydro-1H-isoquinolinyl, thiophenyl, and azepanyl, where each of the rings can be optionally substituted with one or more, preferably one, substituents independently selected from alkyl and halogen. Pyrrolidinyl, pyridinyl, furyl, thiophenyl, and chloro-pyridinyl are particularly preferred.

[0025] The term "carbocyclyl," alone or in combination, refers to a partially unsaturated 4- to 10-membered carbocyclic ring, wherein one or more carbon atoms are optionally substituted with halogen, alkyl, cycloalkyl, alkoxy, oxo, or aryl, preferably alkyl. An example of a carbocyclyl is indanyl.

[0026] The term "amino," alone or in combination, refers to a primary, secondary, or tertiary amino group attached through a nitrogen atom, wherein a secondary amino group has an alkyl or cycloalkyl substituent, and a tertiary amino group has two similar or different alkyl or cycloalkyl substituents, or two nitrogen substituents together form a ring (e.g., -NH, methylamino, ethylamino, dimethylamino, diethylamino, methyl-ethylamino, pyrrolidinyl, and piperidino). Particularly preferred is primary amino.

[0027] The term "cycloalkylalkyl", alone or in combination, denotes an alkyl group substituted with one or more, preferably one, cycloalkyl groups, wherein the terms alkyl and cycloalkyl have the meanings given above.

[0028] The term "cycloalkylalkylcarbonyl," alone or in combination, refers to a cycloalkylalkyl-C(O)- group, where cycloalkylalkyl is defined as above.

[0029] The term "cycloalkylalkoxy", alone or in combination, denotes an alkoxy group substituted with one or more, preferably one, cycloalkyl groups, wherein the terms alkoxy and cycloalkyl have the meanings given above.

[0030] The term "cycloalkylalkoxyalkyl", alone or in combination, denotes an alkyl group substituted with one or more, preferably one, cycloalkylalkoxy groups, wherein the terms alkyl and cycloalkylalkoxy have the meanings given above.

[0031] The term "heterocyclylalkylcarbonyl," alone or in combination, refers to a heterocyclylalkyl-C(O)- group, where heterocyclylalkyl is defined as above.

[0032] The term "aralkylcarbonyl," alone or in combination, refers to an aralkyl-C(O)- group, where aralkyl is defined as above.

[0033] The term "alkylcarbonyl," alone or in combination, refers to an alkyl-C(O)- group, where alkyl is defined as above.

[0034] The term "cycloalkylcarbonyl," alone or in combination, refers to a cycloalkyl-C(O)- group, where cycloalkyl is defined as above.

[0035] The term "arylcarbonyl," alone or in combination, refers to an aryl-C(O)- group, where aryl is defined as above.

[0036] The term "alkoxyalkyl", alone or in combination, denotes an alkyl group substituted with one or more, preferably one, alkoxy groups, wherein the terms alkyl and alkoxy have the meanings given above.

[0037] The term "hydroxyalkyl," alone or in combination, refers to an alkyl group that is substituted with one or more, preferably one, hydroxyl groups, wherein the terms alkyl and hydroxy have the meanings given above.

[0038] The term "heterocyclylalkyl", alone or in combination, denotes an alkyl group substituted with one or more, preferably one, heterocyclyl groups, wherein the terms alkyl and heterocyclyl have the meanings given above.

[0039] The term "heterocyclylcarbonyl," alone or in combination, refers to a heterocyclyl-C(O)- group, where heterocyclyl is defined as above.

[0040] The term "carbocyclylalkyl", alone or in combination, denotes an alkyl group substituted with one or more, preferably one, carbocyclyl groups, wherein the terms alkyl and carbocyclyl have the meanings given above.

[0041] The term "halogen" denotes fluorine, chlorine, bromine or iodine, preferably fluorine, chlorine or bromine, and especially chlorine.

[0042] The compounds of formula I may also be solvated, e.g., hydrated. Solvation may occur during the manufacturing process or may occur, for example, as a result of the hygroscopic properties of the initially anhydrous compound (of formula I) (hydration). The term pharmaceutically acceptable salt also includes pharmaceutically usable solvates.

[0043] More specifically, for example, the -COOH group of the compound according to Formula I can be esterified. The alkyl and aralkyl esters mentioned above are examples of suitable esters. Methyl, ethyl, propyl, butyl, and benzyl esters are preferred esters. Methyl and ethyl esters are particularly preferred.

[0044] Further examples of suitable pharmaceutically acceptable esters are compounds of formula I, in which hydroxyl groups can be esterified. Examples of such esters are formate, acetate, propionate, butyrate, isobutyrate, valerate, 2-methylbutyrate, isovalerate and N,N-dimethylaminoacetate. Preferred esters are acetate and N,N-dimethylaminoacetate.

[0045] "Heteroaryl" refers to an aromatic ring containing at least one heteroatom, such as N, S, O, or Se. The heteroaryl in the present disclosure can be any heteroaryl. The heteroaryl in the present disclosure can be, but is not limited to, pyrrolidinyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, chromanyl, indolinyl, isoindolinyl, furanyl, pyrrolidinyl, pyridinyl, pyrazinyl, pyrimidinyl, triazinyl, thiophenyl, tetrahydrofuranyl, pyrrolyl, oxazolyl, oxadiazolyl, imidazolyl, triazolyl, tetrazolyl, benzoxazolinyl, benzothiazolinyl, benzimidazolidinyl group, or any combination thereof.

[0046] The term "halo," "halogen," or "halide" group, as used herein, alone or as part of another substituent, means, unless otherwise stated, a fluorine, chlorine, bromine, or iodine atom. The compounds described herein may contain one or more chiral centers or may otherwise exist as multiple stereoisomers. In one embodiment, the invention described herein is not limited to any particular stereochemical requirement, and it should be understood that the compounds, as well as compositions, methods, uses, and medicaments comprising them, may be optically pure or may be any of various stereoisomeric mixtures (including racemic mixtures and other mixtures of enantiomers, other mixtures of diastereomers, etc.). It should also be understood that such mixtures of stereoisomers may contain a single stereochemical configuration at one or more chiral centers, as well as a mixture of stereochemical configurations at one or more other chiral centers.

[0047] Similarly, the compounds described herein may contain geometric centers (e.g., cis, trans, E, and Z double bonds). In another embodiment, it should be understood that the invention described herein is not limited to any particular geometric isomer requirement, and that the compounds, and compositions, methods, uses, and medicaments comprising them, may be pure or any of various geometric isomeric mixtures. It should also be understood that such mixtures of geometric isomers may contain a single configuration at one or more double bonds, as well as mixtures of geometries at one or more other double bonds.

[0048] The term "optionally substituted" or "optionally substituents," as used herein, means that the group in question is either unsubstituted or substituted with one or more of the specified substituents. When the group in question is substituted with more than one substituent, the substituents may be the same or different. When the term "independently" is used, "independently are" and "independently selected from" mean that the groups in question may be the same or different. Certain of the terms defined herein may occur more than one time in the structures, and upon such occurrence, each term shall be defined independently of the other.

[0049] The present invention relates to a novel compound that can inhibit the activity of LYP.Therefore, the compound according to the present invention can be used to treat diseases or disorders associated with PTPN22 gene polymorphism, including type 1 diabetes, rheumatoid arthritis, systemic lupus erythematosus, Graves' disease, Addison's disease, vitiligo, juvenile arthritis, Hashimoto's thyroiditis, and other rare diseases.In addition, these LYP inhibitors can be useful in a new class of cancer immunotherapy.The compound can be injected or orally administered.

[0050] Experimental Section

[0051] The present invention also includes processes for preparing the compounds of the present invention. In the reactions described, it may be necessary to protect reactive functional groups (e.g., hydroxy, amino, imino, thio, or carboxy groups, which are desired in the final product) to prevent unwanted participation in the reaction. Conventional protecting groups can be used in accordance with standard practice (see, for example, TW Greene and PGM Wuts in "Protective Groups in Organic Chemistry", John Wiley and Sons, 1991).

[0052] General synthetic procedures and reagents: Unless otherwise specified, all reagents were purchased from commercial suppliers and used directly without further purification. Analytical thin layer chromatography (TLC) was performed on 0.25 mm silica gel 60-F. 254 Column chromatography was performed using KP-SIL silica gel (Biotage, USA), and flash column chromatography was performed on Biotage pre-packed columns using an automated flash chromatography system, Biotage Isolera One. 1 H and 13 C NMR spectra were recorded on a Bruker AVANCE 500 MHz instrument. Proton magnetic resonance spectra ( 1 H NMR chemical shifts were quoted in parts per million (ppm) referenced to the appropriate solvent peak or 0.0 ppm for tetramethylsilane (TMS). The following abbreviations were used to describe peak splitting patterns, where appropriate: br = broad, s = singlet, d = doublet, t = triplet, q = quartet, m = multriplet, dd = doublet of doublet. Coupling constants, J, were reported in hertz (Hz). C NMR chemical shifts were reported in ppm referenced to the center line at 39.52 for DMSO-d. Low-resolution mass spectra and purity data were obtained using an Agilent Technologies 6470 series triple quadrupole LC / MS. High-resolution mass spectra (HRMS) were recorded on an Agilent mass spectrometer using ESI-TOF (electrospray ionization-time of flight).

[0053] Compounds of Formula I were prepared according to the following general synthesis as illustrated in Scheme 1. Where appropriate, protecting groups are used as needed according to established synthetic procedures known to those skilled in the art and may or may not be removed upon completion of the synthesis. Starting materials are synthesized according to methods known in the art or are commercially available. [ka] Scheme 1: General synthesis of compounds of formula I

[0054] Example 1: (S)-6-(2-([1,1'-biphenyl]-4-ylcarboxamido)propanamido)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid [ka]

[0055] Step A and Step B: Ethyl 6-nitro-4-oxo-1,4-dihydroquinoline-3-carboxylate

[0056] Starting from p-nitroaniline and diethyl ethoxymethylenemalonate, the advanced intermediate ethyl 6-nitro-4-oxo-1,4-dihydroquinoline-3-carboxylate was obtained according to a previously reported protocol. See Al-As'ad, RM, El-abadelah, MM, Sabri, SS, Zahra, JA & Voelter, W. Synthesis of 6-Ethyl-1,2,9-trioxopyrrolo[3,2-f]quinoline-8-carboxylic Acid. Z Naturforsch B 68, 700-706, doi:10.5560 / Znb.2013-3009 (2013).

[0057] Step C: Ethyl 6-amino-4-oxo-1,4-dihydroquinoline-3-carboxylate

[0058] To a solution of 6-nitro-4-oxo-1,4-dihydroquinoline-3-carboxylate (2.0 g, 7.63 mmol) in dimethylformamide (DMF, 40 ml) was added 10% Pd / C (0.2 g). Hydrogenation was carried out at 100° C. under 1 atm pressure. After stirring for 12 hours, removal of the catalyst and solvent gave a solid residue, which was then washed with ethyl acetate (40 ml) to give ethyl 6-amino-4-oxo-1,4-dihydroquinoline-3-carboxylate (1.5 g, 85% yield). 1H NMR (500 MHz, DMSO) δ12.03 (s, 1H), 8.32 (s, 1H), 7.33 (d, J = 8.7 Hz, 1H), 7.26 (d, J= 2.6 Hz, 1H), 6.99 (dd, J = 8.7, 2.6 Hz, 1H), 5.45 (s, 2H), 4.18 (q, J= 7.1 Hz, 2H), 1.26 (t, J = 7.1 Hz, 3H). LC-MS (ESI): C 12 H 13 m / z of N2O3 [M + H] + Calculated value: 233.09, Measured value: 233.10.

[0059] Step D: (S)-ethyl 6-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-oxo-1,4-dihydroquinoline-3-carboxylate

[0060] Fmoc-L-Ala-OH (2.0 g, 6.42 mmol), HOBt (1.13 g, 8.35 mmol), and HBTU (3.17 g, 8.35 mmol) were dissolved in dry dimethylformamide (DMF, 40 ml). The mixture was stirred at room temperature for 15 minutes. Ethyl 6-amino-4-oxo-1,4-dihydroquinoline-3-carboxylate (1.34 g, 5.78 mmol) and N,N-diisopropylethylamine (3.4 ml, 19.27 mmol) were then added, and the resulting mixture was stirred at room temperature overnight. DMF was removed by rotary evaporation, and then ethyl acetate and water were added. The formed precipitate was collected by filtration and purified by column chromatography eluting with dichloromethane / methanol 10:1 v / v to give the Fmoc-protected intermediate as a light brown solid (2.2 g, 65% yield). 1H NMR (500 MHz, DMSO) δ 12.28 (d, J = 6.4 Hz, 1H), 10.27(s, 1H), 8.47 (d, J = 6.5 Hz, 1H), 8.39 (d, J = 2.1 Hz, 1H), 7.95(dd, J = 8.9, 2.3 Hz, 1H), 7.88 (d, J = 7.6 Hz, 2H), 7.75 - 7.68(m, 3H), 7.57 (d, J = 8.9 Hz, 1H), 7.44 - 7.37 (m, 2H), 7.35 - 7.28 (m,2H), 4.28 - 4.26 (m, 2H), 4.21 - 4.17 (m, 4H), 1.32 (d, J = 7.1 Hz, 3H),1.26 (t, J = 7.1 Hz, 3H). LC-MS (ESI): C 30 H 28 m / z of N3O6 [M + H] + Calculated value: 526.20, Measured value: 526.30.

[0061] Step E: (S)-Ethyl 6-(2-aminopropanamido)-4-oxo-1,4-dihydroquinoline-3-carboxylate

[0062] The Fmoc-protected intermediate (S)-ethyl 6-(2-((((9H-fluoren-9-yl)methoxy)carbonyl)amino)propanamido)-4-oxo-1,4-dihydroquinoline-3-carboxylate (2.1 g, 4.0 mmol) was dissolved in DMF (30 ml). Piperidine (7.5 ml) was added and the reaction mixture was stirred at room temperature for 1 hour. Concentration in vacuo gave a brown solid which was washed with ethyl acetate to give the title compound (0.83 g, 69% yield). 1 H NMR (500 MHz, DMSO) δ 8.49 (s, 1H),8.43 (d, J = 2.4 Hz, 1H), 7.98 (dd, J = 8.9, 2.4 Hz, 1H), 7.58(d, J = 8.9 Hz, 1H), 4.21 (q, J = 7.1 Hz, 2H), 3.52 - 3.47 (m,1H), 1.27 (t, J = 7.1 Hz, 3H), 1.25 (d, J = 6.9 Hz, 3H). LC-MS(ESI): C 15 H 18 m / z of N3O4 [M + H] + Calculated value: 304.13, Measured value: 304.20.

[0063] Step F: (S)-ethyl 6-(2-([1,1'-biphenyl]-4-ylcarboxamido)propanamido)-4-oxo-1,4-dihydroquinoline-3-carboxylate

[0064] Biphenyl-4-carboxylic acid (0.40 g, 2.02 mmol), HOBt (0.35 g, 2.62 mmol), and HBTU (1.0 g, 2.62 mmol) were dissolved in dry dimethylformamide (DMF, 20 ml). The mixture was stirred at room temperature for 15 minutes. (S)-ethyl 6-(2-aminopropanamido)-4-oxo-1,4-dihydroquinoline-3-carboxylate (0.55 g, 1.82 mmol) and N,N-diisopropylethylamine (1.07 ml, 6.05 mmol) were then added, and the resulting mixture was stirred at room temperature overnight. DMF was removed by rotary evaporation, and then ethyl acetate and water were added. The formed precipitate was collected by filtration and washed with ethyl acetate to give (S)-ethyl 6-(2-([1,1′-biphenyl]-4-ylcarboxamido)propanamido)-4-oxo-1,4-dihydroquinoline-3-carboxylate (0.76 g, 78% yield). 1 H NMR (500 MHz, DMSO) δ12.30 (d, J = 6.7 Hz, 1H), 10.36 (s, 1H), 8.74 (d, J = 7.0 Hz,1H), 8.48 (d, J = 6.7 Hz, 1H), 8.41 (d, J = 2.3 Hz, 1H), 8.03 (d,J = 8.4 Hz, 2H), 8.00 (dd, J = 8.9, 2.4 Hz, 1H), 7.79 (d, J= 8.4 Hz, 2H), 7.75 (d, J = 7.2 Hz, 2H), 7.59 (d, J = 8.9 Hz,1H), 7.50 (t, J = 7.6 Hz, 2H), 7.41 (t, J = 7.4 Hz, 1H), 4.67 -4.61 (m, 1H), 4.21 (q, J = 7.1 Hz, 2H), 1.48 (d, J = 7.2 Hz, 3H),1.28 (t, J = 7.1 Hz, 3H). LC-MS (ESI): C 28 H 26 m / z of N3O5 [M + H] + Calculated value: 484.19, Measured value: 484.20.

[0065] Step G: (S)-6-(2-([1,1'-biphenyl]-4-ylcarboxamido)propanamido)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid (Example 1)

[0066] To a solution of the compound (S)-ethyl 6-(2-([1,1'-biphenyl]-4-ylcarboxamido)propanamido)-4-oxo-1,4-dihydroquinoline-3-carboxylate (500 mg, 1.03 mmol) in methanol (20 ml) and HO (20 ml) was added KOH (580 mg, 10.34 mmol). The resulting mixture was stirred at 60°C for 16 hours. The mixture was brought to 0°C and carefully acidified with 1N HCl to pH = 1. The formed precipitate was collected by filtration and purified by HPLC to give the desired product as an off-white solid (422 mg, 90% yield). 1 H NMR (500 MHz, DMSO) δ 10.53 (s,1H), 8.83 (d, J = 6.8 Hz, 1H), 8.78 (d, J = 6.8 Hz, 1H), 8.65 (d,J = 2.4 Hz, 1H), 8.10 (dd, J = 9.1, 2.4 Hz, 1H), 8.04 (d, J= 8.5 Hz, 2H), 7.84 - 7.77 (m, 3H), 7.76 - 7.70 (m, 2H), 7.50 (t, J =7.6 Hz, 2H), 7.45 - 7.39 (m, 1H), 4.68 - 4.61 (m, 1H), 1.49 (d, J = 7.2Hz, 3H). 13C NMR (126 MHz, DMSO) δ 177.99 (s), 172.03 (s), 166.55(s), 166.08 (s), 144.05 (s), 142.89 (s), 139.15 (s), 137.34 (s), 135.40 (s),132.67 (s), 129.04 (s), 128.30 (s), 128.08 (s), 126.88 (s), 126.41 (s), 126.19(s), 124.95 (s), 120.41 (s), 113.21 (s), 107.07 (s), 50.13 (s), 17.61 (s).LC-MS (ESI): C 26 H 20 m / z of N3O5 [M - H] - Calculated: 454.14, Found: 454.30. HRMS (ESI-TOF): C 26 H 20 m / z of N3O5 [M - H] - Calculated: 454.1403, Found: 454.1413; Purity: >95% (UV, λ = 254 nm).

[0067] Examples 2-51 were prepared according to the procedure described above for Example 1. All Examples 1-51 are listed in Table 1. Table 1: Examples 1 to 51 [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6] [Table 1-7] [Table 1-8]

[0068] Example 52: (S)-6-(2-([1,1'-biphenyl]-4-ylcarboxamido)propanamido)-1-(3,4-dichlorobenzyl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid [ka] Scheme 2. Synthesis of Example 52

[0069] Process A: (S)-Ethyl 6-(2-([1,1'-biphenyl]-4-ylcarboxamido)propanamido)-1-(3,4-dichlorobenzyl)-4-oxo-1,4-dihydroquinoline-3-carboxylate

[0070] To a solution of the compound (S)-ethyl 6-(2-([1,1'-biphenyl]-4-ylcarboxamido)propanamido)-4-oxo-1,4-dihydroquinoline-3-carboxylate (200 mg, 0.41 mmol) in DMF (20 ml), K2CO3 (172 mg, 1.24 mmol) and 4-(bromomethyl)-1,2-dichlorobenzene (110 mg, 0.45 mmol) were added. The solution was maintained at 80 °C for 2 hours. After cooling to room temperature, the excess K2CO3 was then filtered and the solvent was evaporated. The oily residue was purified by column to give the desired product as an off-white solid (232 mg, 87% yield). LC-MS (ESI): C 35 H 30 m / z of Cl2N3O5 [M + H] + Calculated value: 642.16, Measured value: 642.10.

[0071] Step B: (S)-6-(2-([1,1'-biphenyl]-4-ylcarboxamido)propanamido)-1-(3,4-dichlorobenzyl)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid

[0072] To a solution of the compound (S)-ethyl 6-(2-([1,1'-biphenyl]-4-ylcarboxamido)propanamido)-1-(3,4-dichlorobenzyl)-4-oxo-1,4-dihydroquinoline-3-carboxylate (100 mg, 0.15 mmol) in methanol (4 ml) and HO (4 ml) was added KOH (87 mg, 1.56 mmol). The resulting mixture was stirred at 60°C for 16 hours. The mixture was brought to 0°C and carefully acidified with 1N HCl to pH = 1. The formed precipitate was collected by filtration and purified by HPLC to give the desired product as an off-white solid (81 mg, 84% yield). LC-MS (ESI): C 33 H 24 m / z of Cl2N3O5 [M - H] - Calculated: 612.11, Found: 612.20. HRMS (ESI-TOF): C 26 H 20 m / z of N3O5 [M - H] - Calculated: 612.1093, Found: 612.1101; Purity: >95% (UV, λ = 254 nm).

[0073] Examples 53-74 were prepared according to the procedure described above for Example 52. Examples 52-74 are all listed in Table 2. Table 2: Examples 52-74 [Table 2-1] [Table 2-2] [Table 2-3] [Table 2-4]

[0074] Compounds of formula I, where X is H, were prepared according to the following general synthetic scheme 3. Where appropriate, protecting groups are used as needed according to established synthetic procedures known to those skilled in the art and may or may not be removed upon completion of the synthesis. Starting materials are synthesized according to methods known in the art or are commercially available. [ka] Scheme 3. Method for synthesizing compounds of formula I where X is H

[0075] Example 75: (S)-6-(2-([1,1'-biphenyl]-4-ylcarboxamido)propanamido)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid [ka]

[0076] Process A: (S)-Ethyl 6-((2-((tert-butoxycarbonyl)amino)propyl)amino)-4-oxo-1,4-dihydroquinoline-3-carboxylate

[0077] A mixture of ethyl 6-amino-4-oxo-1,4-dihydroquinoline-3-carboxylate (200 mg, 0.86 mmol), (S)-tert-butyl (1-oxopropan-2-yl)carbamate (150 mg, 0.86 mmol), and acetic acid (1 mL) in anhydrous methanol (15 mL) was refluxed with stirring for 16 hours. After cooling, NaBHCN (109 mg, 1.72 mmol) was added, and the mixture was stirred in an ice-water bath for 1 hour and then refluxed for an additional 3 hours. The solvent was evaporated, and methanol (15 mL) was added to the residue again. The formed precipitate was collected by filtration and purified by column chromatography eluting with dichloromethane / methanol 10:1 v / v to give the Boc-protected intermediate (S)-ethyl 6-((2-((tert-butoxycarbonyl)amino)propyl)amino)-4-oxo-1,4-dihydroquinoline-3-carboxylate as a light brown solid (320 mg, 95% yield). LC-MS (ESI): C 20 H 28 m / z of N3O5 [M + H] + Calculated value: 390.20, Measured value: 390.30.

[0078] Step B: (S)-Ethyl 6-((2-aminopropyl)amino)-4-oxo-1,4-dihydroquinoline-3-carboxylate

[0079] The above Boc-protected intermediate (S)-ethyl 6-((2-((tert-butoxycarbonyl)amino)propyl)amino)-4-oxo-1,4-dihydroquinoline-3-carboxylate (250 mg, 0.64 mmol) was dissolved in DCM (20 ml). CF3COOH (5 ml) was added and the reaction mixture was stirred at room temperature for 1 hour. Concentration in vacuo gave a brown solid, which was dissolved in ethyl acetate and washed with aqueous NaHCO3. The organic solvents were collected and concentrated in vacuo to give a brown solid, which was then purified by column chromatography eluting with dichloromethane / methanol 10:1 v / v to give the title compound (170 mg, 91% yield). LC-MS (ESI): C 15 H 20 m / z of N3O3 [M + H] + Calculated value: 290.15, Measured value: 290.20.

[0080] Step C: (S)-ethyl 6-((2-([1,1'-biphenyl]-4-ylcarboxamido)propyl)amino)-4-oxo-1,4-dihydroquinoline-3-carboxylate

[0081] Biphenyl-4-carboxylic acid (50 mg, 0.25 mmol), HOBt (38 mg, 0.28 mmol), and HBTU (106 mg, 0.28 mmol) were dissolved in dry dimethylformamide (DMF, 10 ml). The mixture was stirred at room temperature for 15 minutes. (S)-ethyl 6-((2-aminopropyl)amino)-4-oxo-1,4-dihydroquinoline-3-carboxylate (73 mg, 0.25 mmol) and N,N-diisopropylethylamine (0.134 ml, 0.76 mmol) were then added, and the resulting mixture was stirred at room temperature overnight. DMF was removed by rotary evaporation, and then ethyl acetate and water were added. The formed precipitate was collected by filtration and washed with ethyl acetate to give (S)-ethyl 6-((2-([1,1'-biphenyl]-4-ylcarboxamido)propyl)amino)-4-oxo-1,4-dihydroquinoline-3-carboxylate (85 mg, 71% yield). LC-MS (ESI): C 28 H 28 m / z of N3O4 [M + H] + Calculated value: 470.21, Measured value: 484.20.

[0082] Step D: (S)-6-((2-([1,1'-biphenyl]-4-ylcarboxamido)propyl)amino)-4-oxo-1,4-dihydroquinoline-3-carboxylic acid

[0083] To a solution of compound (S)-ethyl 6-((2-([1,1'-biphenyl]-4-ylcarboxamido)propyl)amino)-4-oxo-1,4-dihydroquinoline-3-carboxylate (80 mg, 0.17 mmol) in methanol (4 ml) and HO (4 ml) was added KOH (116 mg, 2.06 mmol). The resulting mixture was stirred at 60°C for 16 hours. The mixture was brought to 0°C and carefully acidified with 1N HCl to pH = 1. The formed precipitate was collected by filtration and purified by HPLC to give the desired product, Example 75, as an off-white solid (58 mg, 77% yield). LC-MS (ESI): C 26H 22 m / z of N3O4 [M - H] - Calculated: 440.16, Found: 440.20. Purity: >95% (UV, λ=254 nm).

[0084] Examples 76-98 were prepared according to the procedure described above for Example 75.

[0085] Table 3: Examples 75-98 [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4]

[0086] Assay

[0087] Compounds of the present invention were assayed for their ability to selectively inhibit LYP activity. The inhibitory properties of the compounds of the invention described herein may be demonstrated by testing them in any one of the following assays.

[0088] Expression and purification of the LYP catalytic domain

[0089] The N-terminal (His)6-tagged LYP catalytic domain (residues 1-303) was subcloned into pET28a. For protein expression, the LYP expression construct was transformed into Escherichia coli BL21-(DE3). Transformed cells were grown in Luria broth (LB) containing 100 μg / mL ampicillin for 4 hours at 37°C until an OD600 of 0.6 was reached, and then induced with 0.4 mM IPTG for overnight growth at room temperature. Cells were harvested by centrifugation (6000 rpm, 15 minutes at 4°C), and the cell pellet from 1.5 L of LB medium was suspended in 30 mL of ice-cold lysis buffer consisting of 5 mM imidazole, 500 mM NaCl, 20 mM Tris-HCl (pH 7.9), 0.05 mg / mL trypsin inhibitor, and 0.1 mM PMSF. The suspension was passed twice through a French press at 1000 psi, and the cell lysate was centrifuged at 15,000 rpm for 30 minutes at 4°C. The supernatant was mixed with 2 mL of Ni-NTA agarose (His*Bind Resin) (Qiagen) for 1 hour at 4°C, and the mixture was then transferred to an empty column. The column was washed with 200 mL of binding buffer (5 mM imidazole, 500 mM NaCl, 20 mM Tris-HCl (pH 7.9)), followed by 20 mL of wash buffer (20 mM imidazole, 500 mM NaCl, 20 mM Tris-HCl (pH 7.9)), and then eluted with 20 mL of elution buffer (200 mM imidazole, 500 mM NaCl, 20 mM Tris-HCl (pH 7.9), 5 mM DTT). The eluate was dialyzed against 1 L of buffer A (50 mM NaCl, 20 mM MES (pH 5.8), 1 mM EDTA) for 6 hours at 4°C and then loaded onto a Mono S column equilibrated with buffer A at 4°C. The column was washed with 10 mL of buffer A and then eluted with a 40 mL linear gradient of 0 to 1 M NaCl in buffer A. The column fractions were analyzed by measuring absorbance at 280 nm and by SDS-PAGE analysis.The fractions were combined and concentrated to ≦1 mL using an Amicon concentrator at 4° C., then loaded onto a gel filtration column, Superdex 75. The column was eluted with buffer A, and the protein-containing fractions were combined, concentrated to 8 mg / mL, and stored at −80° C. The LYP preparation was shown to be homogeneous by SDS-PAGE analysis.

[0090] Enzyme kinetics assay

[0091] PTP activity was assayed using p-nitrophenyl phosphate (pNPP) as a substrate in 3,3-dimethylglutarate buffer (50 mM 3,3-dimethylglutarate, pH 7.0, 1 mM EDTA, 150 mM NaCl) at 25°C. The assay was performed in a 96-well plate. Typically, the IC of LYP was 50 To determine the K value, reactions were run at 5.0 mM (K of the above substrates relative to Lyp) using serial dilutions. m The reaction was initiated by adding the enzyme (20 nM final concentration) to the reaction mixture (0.2 mL) containing pNPP. 50 To determine the K values, repeat the assay described above and compare the concentration of pNPP with the corresponding K value for each PTP. m The experiments were carried out under the same conditions as those used for LYP, except that the concentration of the compound used to determine the IC50 value was set to IC. All PTPs used in this study were recombinant proteins prepared in-house. The concentration of compound used to determine the IC50 value was set to IC 50 The values ​​ranged from 0.2-fold to 5-fold of the original values. The reaction rates were measured using a SpectraMax Plus 384 microplate spectrophotometer (Molecular Devices). To determine the mode of inhibition, the reaction was initiated by adding LYP (5 nM final concentration) to a reaction mixture (0.2 mL) containing various concentrations of pNPP and inhibitor L-1 (Example 1). Data were fitted using SigmaPlot Enzyme Kinetics Module (Systat Software, Inc.).

[0092] Pharmacokinetic studies

[0093] Example 1 shows IC values ​​as low as 1.4±0.2 μM. 50 Example 1 is a novel PTPN22 inhibitor having the following structure: To test the efficacy of Example 1 in a mouse model, pharmacokinetic data are required to understand its absorbance / distribution / metabolism / excretion (ADME) properties. The detailed experimental procedures and obtained pharmacokinetic parameters are shown below.

[0094] Animal dosing and sample collection for pharmacokinetic studies

[0095] Example 1 was first dissolved in DMSO to prepare a 20 mg / ml solution. This solution was then further diluted to a 2 mg / ml solution. The formulation was 10% DMSO - 85% PBS - 5% Cremophor EL (CrEL). Each mouse was administered a single IP dose of 10 mg / kg. The volume of each injection was approximately 100 μL according to the mouse's weight. At different time points (1 hour, 1.5 hours, 2 hours, 2.5 hours, 3 hours, 6 hours, and 24 hours), blood samples (50 μL) were collected and centrifuged to obtain serum. Then, the serum (10 μL) was mixed with acetonitrile (20 μL) and centrifuged. The supernatant was collected and subjected to liquid chromatography / mass spectrometry analysis.

[0096] Analysis by liquid chromatography / mass spectrometry

[0097] Liquid chromatography / mass spectrometry (LC / MS) analysis was performed on an Agilent 1260 analytic HPLC system equipped with a Kinetex 2.6 µm C18 column (3 mm x 50 mm) and an Agilent 6470 Triple Quadrupole MS detector. The elution was performed with 0-100% MeOH-HO with 0.1% (w / v) formic acid at a flow rate of 0.7 mL / min (gradient: 1.2 min 0-10% MeOH linear gradient, 1.5 min 10-90% MeOH linear gradient, followed by 1.3 min 90-100% MeOH, followed by 2.5 min 100% MeOH). The MS detector was set to single ion mode (SIM) to monitor the negative charge of 454.2 (M-1). The detection limit for Example 1 is 100 nM with a 4 µL sample injection.

[0098] Data analysis

[0099] Pharmacokinetic parameters were calculated in GraphPad Prism 6 and the results are shown below in Table 4. The pharmacokinetic curves of Example 1 are shown in Figures 3 and 4.

[0100] Table 4. Pharmacokinetic data for Example 1 [Table 4]

[0101] LYP inhibitor, Example 1, shows potent in vivo activity in a mouse model of cancer immunotherapy

[0102] Figure 5 illustrates that Example 1 demonstrates effective in vivo activity in a mouse model of cancer immunotherapy. As observed in the PTPN22(LYP) KO model, treatment of WT mice with Example 1 (a) resulted in significantly reduced MC38 tumor growth compared to the vehicle-injected control group (b). We further tested the effect of Example 1 on another syngeneic immunocompetent model, CT26, in Balb / c mice, demonstrating similar antitumor effects (c). Analysis of MC38 tumors by immunohistochemistry showed that Example 1 induced increased infiltration of CD4+ and CD8+ T cells (d). Furthermore, profiling of the immune infiltrates of both MC38 and CT26 tumors demonstrated significantly improved presence of multiple immune cell types and T cell subtypes in tumors treated with Example 1. To test whether the effects of Example 1 treatment could be due to off-target effects, we treated MC38 tumors in PTPN22 KO mice with injections of either vehicle or Example 1. No significant difference in tumor growth was observed, suggesting that the protective effect of Example 1-mediated tumor growth was PTPN22(LYP)- and host-specific (e).

[0103] In conclusion, Example 1 exhibits strong in vivo antitumor activity that closely phenocopies the PTPN22 KO mouse, with little off-target activity.

[0104] In one embodiment, the present disclosure provides a compound of formula I: [ka] or a stereoisomer, tautomer, solvate, derivative, or pharmaceutically acceptable salt thereof, wherein: R 1 and R 2independently represent hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, alkylcarbonyl, provided that said alkylcarbonyl is not methylcarbonyl, optionally substituted cycloalkylcarbonyl, optionally substituted cycloalkylalkylcarbonyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted arylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted alkoxyalkyl, optionally substituted hydroxyalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylcarbonyl, optionally substituted heterocyclylalkylcarbonyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted amino, optionally substituted alkyl-SO2-, optionally substituted aryl-SO2-, optionally substituted heterocyclyl-SO2-, optionally substituted amino-SO2-; 1 and R 2 together with the N atom to which they are attached form a 5-10 membered heterocyclic ring optionally containing a second heteroatom selected from nitrogen or oxygen, wherein said heterocyclic ring is optionally substituted; R 3 is H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heterocyclyl; R 4 and R 6 represent independently hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and heteroaryl; optionally substituted aralkyl, or optionally substituted heteroaralkyl; (R 5 ) n when n is 1 or 2, represents 1 to 2 of -H, -F, -Cl, -Br, -I, -CFH2, -CF2H, -CF3, -CN, -OH, -NO2, -NH2, -SO2CH3, SO2NH2, -SON2NHCH3, optionally substituted -CO2-alkyl, optionally substituted NH(alkyl) or N(alkyl)2, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkoxy, optionally substituted S-alkyl, optionally substituted cycloalkyl, optionally substituted heterocycle, optionally substituted aryl or optionally substituted heteroaryl; X is O or H, where -C=X- is -CH2- when X is H, where R 1 and R 2 is optionally substituted arylcarbonyl when X is H, and R 1 and R 2 One of them is R 3 to form a ring; Y is CR 5 or N, The present invention provides a compound or a stereoisomer, tautomer, solvate, derivative, or pharmaceutically acceptable salt thereof.

[0105] In one embodiment of the present disclosure relating to compounds of formula I, X is O.

[0106] In one embodiment of the present disclosure relating to compounds of formula I, X is H to make -C=X- a CH2 group.

[0107] In one embodiment of the present disclosure relating to compounds of formula I, Y is CH, CF, C—Cl, C—Br, CI, or N.

[0108] In one embodiment of the present disclosure relating to compounds of formula I, R 4 is H.

[0109] In one embodiment of the present disclosure relating to compounds of formula I, R 6 is H.

[0110] In one embodiment of the present disclosure relating to compounds of formula I, R 3 is an optionally substituted C1-C4 alkyl.

[0111] In one embodiment of the present disclosure relating to compounds of formula I, R 4 and R 6 is H and R 3 is an optionally substituted C1-C4 alkyl.

[0112] In one embodiment of the present disclosure relating to compounds of formula I, R 1 and R 2 At least one of the groups is H.

[0113] In one embodiment of the present disclosure relating to compounds of formula I, R 4 and R 6 is H and R 3 is optionally substituted C1-C4 alkyl, and R 1 and R 2 At least one of the groups is H.

[0114] In one embodiment of the present disclosure relating to compounds of formula I, R 1 and R 2 one of R is optionally substituted arylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted heterocyclylcarbonyl, or optionally substituted heterocyclylalkylcarbonyl; 1 and R 2 One of the is H.

[0115] In one embodiment of the present disclosure relating to compounds of formula I, R 1 and R 2 is an optionally substituted biphenylcarbonyl or phenylcarbonyl, and R 1and R 2 One of the is H.

[0116] In one embodiment of the present disclosure relating to compounds of formula I, R 1 ~R 6 , X, Y, and n are the specific R 1 ~R 6 , can be any combination of X, Y, and n.

[0117] In one embodiment of the present disclosure regarding compounds of Formula I, the COOH group of the compound may form derivatives, including, but not limited to, ester amides and carbamates. The alkyl and aralkyl derivatives are examples of suitable ester amides or carbamates. The methyl, ethyl, propyl, butyl, and benzyl esters are preferred ester amides or carbamates. The methyl and ethyl ester amides or carbamates are particularly preferred. Further examples of pharmaceutically suitable esters are compounds of Formula I in which the hydroxyl group can be esterified. Examples of such esters are formates, acetates, propionates, butyrates, isobutyrates, valerates, 2-methylbutyrates, isovalerates, and N,N-dimethylaminoacetates. Preferred esters are acetates and N,N-dimethylaminoacetates.

[0118] In one embodiment of the present disclosure relating to compounds of formula I, the compound is: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] and stereoisomers, tautomers, solvates, derivatives, and pharmaceutically acceptable salts thereof.

[0119] In one embodiment, the disclosure provides a method of inhibiting LYP in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula I, a therapeutically suitable stereoisomer, tautomer, solvate, or salt thereof.

[0120] In one embodiment, the disclosure provides a method of treating a patient having a disease or disorder associated with a PTPN22 genetic polymorphism (including, but not limited to, type 1 diabetes, rheumatoid arthritis, systemic lupus erythematosus, Graves' disease, Addison's disease, vitiligo, juvenile arthritis, Hashimoto's thyroiditis, and other additional rare diseases) with a therapeutically effective amount of a compound of Formula I, a therapeutically suitable stereoisomer, tautomer, solvate, or salt thereof.

[0121] In one embodiment, the disclosure provides a method of cancer immunotherapy by inhibiting LYP activity, the method comprising administering to the subject a therapeutically effective amount of a compound of Formula I, or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof.

[0122] In one embodiment, the disclosure provides a pharmaceutical composition comprising a compound of Formula I, or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0123] In one embodiment of the disclosure relating to methods of treating a patient to treat a disease or disorder, a compound of formula I, or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof, can be injected or administered orally.

[0124] Those skilled in the art will recognize that many modifications may be made to the specific implementations described above, and that the implementations should not be limited to the specific described limitations. Other implementations may be possible. The present invention provides, for example, the following items. (Item 1) Compounds of Formula I: [ka] or a stereoisomer, tautomer, solvate, derivative, or pharmaceutically acceptable salt thereof, wherein: R 1 and R 2 independently represent hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, alkylcarbonyl, provided that said alkylcarbonyl is methylcarbonyl, optionally substituted cycloalkylcarbonyl, optionally substituted cycloalkylalkylcarbonyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted arylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted alkoxyalkyl, optionally substituted hydroxyalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylcarbonyl, optionally substituted heterocyclylalkylcarbonyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted amino, optionally substituted alkyl-SO 2 -, optionally substituted aryl-SO 2 -, optionally substituted heterocyclyl-SO 2 -, optionally substituted amino-SO 2 -Not or R 1 and R 2 together with the N atom to which they are attached form a 5-10 membered heterocyclic ring optionally containing a second heteroatom selected from nitrogen or oxygen, wherein said heterocyclic ring is optionally substituted; R 3 is H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heterocyclyl; R 4 and R 6 represent independently hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and heteroaryl; optionally substituted aralkyl, or optionally substituted heteroaralkyl; (R 5 ) n When n is 1 or 2, one or two of -H, -F, -Cl, -Br, -I, or -CFH 2 , -CF 2 H, -CF 3 , -CN, -OH, -NO 2 , -NH 2 , -SO 2 CH 3 , SO 2 NH 2 , -SO 2 NHCH 3 , optionally substituted -CO 2 -alkyl, optionally substituted NH(alkyl) or N(alkyl) 2 , optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkoxy, optionally substituted S-alkyl, optionally substituted cycloalkyl, optionally substituted heterocycle, optionally substituted aryl or optionally substituted heteroaryl; X is O or H, where -C=X- represents -CH when X is H. 2 - where R 1 and R 2 is optionally substituted arylcarbonyl when X is H, and R 1 and R 2 One of them is R 3 to form a ring; Y is CR 5 or N, A compound or a stereoisomer, tautomer, solvate, derivative, or pharmaceutically acceptable salt thereof. (Item 2) R 4 and R 6 is H and R 3 is replaced by C 1 -C 4 The compound according to item 1, wherein the aryl group is alkyl. (Item 3) R 1 and R 2 The compound according to item 1, wherein at least one of (Item 4) R 4 and R 6 is H and R 3 is replaced by C 1 -C 4 alkyl, and R 1 and R 2 The compound according to item 1, wherein at least one of (Item 5) R 1 and R 2 one of R is optionally substituted arylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted heterocyclylcarbonyl, or optionally substituted heterocyclylalkylcarbonyl; 1 and R 2 The compound according to item 1, wherein one of (Item 6) R 1 and R 2 is an optionally substituted biphenylcarbonyl or phenylcarbonyl, and R1 and R 2 The compound according to item 1, wherein one of (Item 7) The compound according to item 1, wherein X is O. (Item 8) Item 2. The compound according to item 1, wherein Y is —CH, CF, C—Cl, C—Br, CI, or N. (Item 9) The compound is selected from the group consisting of:

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Claims

1. Compounds of Formula I: 【Chemistry 17】 or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof, wherein: R １ and R ２ are independently hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted cycloalkylalkyl, alkylcarbonyl, optionally substituted cycloalkylcarbonyl, optionally substituted cycloalkylalkylcarbonyl, optionally substituted aryl, optionally substituted aralkyl, optionally substituted arylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted alkoxyalkyl, optionally substituted hydroxyalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heterocyclylcarbonyl, optionally substituted heterocyclylalkylcarbonyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted amino, optionally substituted alkyl-SO ２ -, optionally substituted aryl-SO ２ -, optionally substituted heterocyclyl-SO ２ -, optionally substituted amino-SO ２ -, where R １ and R ２ wherein the alkylcarbonyl is not methylcarbonyl, or R １ and R ２ together with the N atom to which they are attached form a 5-10 membered heterocyclic ring optionally containing a second heteroatom selected from nitrogen or oxygen, wherein said heterocyclic ring is optionally substituted; R ３ is H, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted cycloalkyl, optionally substituted aryl, optionally substituted heteroaryl, or optionally substituted heterocyclyl; R ４ and R ６ independently represent hydrogen, optionally substituted alkyl, optionally substituted cycloalkyl, optionally substituted heterocycloalkyl, optionally substituted aryl and heteroaryl; optionally substituted aralkyl, or optionally substituted heteroaralkyl; n is 1 or 2; R ５ is -H, -F, -Cl, -Br, -I, -CFH ２ , -CF ２ H, -CF ３ , -CN, -OH, -NO ２ , -NH ２ , -SO ２ CH ３ , S.O. ２ NH ２ , -SO ２ NHCH ３ , optionally substituted —CO ２ -alkyl, optionally substituted NH(alkyl) or N(alkyl) ２ , optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkoxy, optionally substituted S-alkyl, optionally substituted cycloalkyl, optionally substituted heterocycle, optionally substituted aryl or optionally substituted heteroaryl; X is O or -C(=X)- is -CH ２ - and R １ and R ２ At least one of the groups -C(=X)- is -CH ２ - is an optionally substituted arylcarbonyl when -, and one of R 1 and R 2 can be joined to R 3 to form a ring when -C(=X)- is -CH 2 -; Y is C-R ５ or N, A compound or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof.

2. R ４ and R ６ is H and R ３ is an optionally substituted C １ -C ４ The compound of claim 1 , wherein the aryl group is alkyl.

3. R １ and R ２ and n is 0 or 1. The compound of claim 1, wherein at least one of

4. R ４ and R ６ is H and R ３ is an optionally substituted C １ -C ４ alkyl, and R １ and R ２ and n is 0 or 1. The compound of claim 1, wherein at least one of

5. R １ and R ２ one of which is optionally substituted arylcarbonyl, optionally substituted aralkylcarbonyl, optionally substituted heterocyclylcarbonyl, or optionally substituted heterocyclylalkylcarbonyl; R １ and R ２ 10. The compound of claim 1, wherein one of

6. R １ and R ２ is an optionally substituted biphenylcarbonyl or phenylcarbonyl, and R １ and R ２ 10. The compound of claim 1, wherein one of

7. 2. The compound of claim 1, wherein X is O.

8. 2. The compound of claim 1, wherein Y is —C—H, C—F, C—Cl, C—Br, C—I, or N. 【Request Item 9】 【Chemistry 18】 【Chemistry 19】 【Chemistry 20】 【Chemistry 21】 【Chemistry 22】 【Chemistry 23】 【Chemistry 24】 【Chemistry 25】 【Chemistry 26】 and any stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof.

10. 10. A composition for treating a patient having a disease, comprising the compound of claim 1, or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof.

11. 11. The composition of claim 10, wherein the disease comprises type 1 diabetes, rheumatoid arthritis, systemic lupus erythematosus, Graves' disease, Addison's disease, vitiligo, juvenile arthritis, Hashimoto's thyroiditis, or any combination thereof.

12. A composition for treating a patient in need of cancer immunotherapy associated with the activity of LYP, comprising the compound of claim 1, or a stereoisomer, tautomer, solvate, or pharmaceutically acceptable salt thereof.

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