Inhibitors of protein tyrosine phosphatase, compositions, and methods of use

Abstract

This invention discloses compounds of formula (I); pharmaceutically acceptable salts thereof as defined herein, pharmaceutical compositions thereof, and combinations thereof, and methods of using them as inhibitors of protein tyrosine phosphatase (PTPN2). These compounds are suitable for treating cancers and diseases susceptible to PTPN2 inhibition.

Description

Technical Field

[0001] This invention discloses compounds, their pharmaceutically acceptable salts, their pharmaceutical compositions and combinations thereof, and methods of using them as inhibitors of protein tyrosine phosphatase. Prior Technology

[0002] Immune checkpoint blockade (ICB) is a novel approach to immunotherapy that targets immune evasion mechanisms to improve clinical responses in cancer patients. For example, in the treatment of various types of cancer, checkpoint blockade antibodies target cytotoxic T-lymphocyte antigen 4 (CTLA-4), programmed cell death 1 (PD-1), and their ligands, such as programmed cell death ligand 1 (PD-L1), to significantly improve treatment and survival outcomes in patients affected by these malignancies.

[0003] However, most patients who develop ICB are difficult to treat or eventually develop resistance. Specifically, mutations or loss of the interferon-γ (IFNγ) signaling pathway represent an important mechanism of clinical ICB resistance (Zaretsky, N. Engl. J. Med. 375, 819-829). IFNγ is a T cell-derived cytokine that directly restricts tumor growth via Janus kinase / signal transducer and transcription pathway activator (JAK / STAT) signaling. Furthermore, IFNγ indirectly restricts tumor growth by promoting the upregulation of major histocompatibility complex class I (MHC-I), thereby enabling antigen (Ag) presentation to T cells. In vivo CRISPR screening using syngeneic mouse models has revealed the enrichment of the IFNγ pathway in tumors resistant to PD-1. These studies identified the aforementioned IFNγ pathway members (JAK1 / 2 and STAT1) and interferon γ receptors (IFNGR1 / IFNGR2) as resistance targets, as well as newly identified negative regulators (such as PTPN2 and APLNR, which represent novel therapeutic targets (Charles Sinclair et al., Emerg Top Life Sci. (2021) 5 (5): 675-680).

[0004] Data collected from in vivo gene screening using CRISPR-Cas9 genome editing to identify genes resistant to checkpoint blockade indicate that deletion of the protein tyrosine phosphatase (PTPN2) gene in tumor cells increases the efficacy of immunotherapy. The PTPN2 gene encodes a protein tyrosine phosphatase that regulates a range of intracellular processes. Loss of PTPN2 in tumor cells promotes the amplification of IFNγ signaling, antigen presentation to T cells, and growth arrest in response to cytokines; these data suggest that therapeutic inhibition of PTPN2 can enhance the effects of immunotherapies that invoke IFNγ responses (Manguso, Robert T et al., Nature Vol. 547, 7664 (2017): 413-418).

[0005] Protein tyrosine phosphatase non-receptor type 2 (PTPN2) (also known as T-cell protein tyrosine phosphatase (TCPTP)) is an intracellular member of the first subfamily of phosphate-tyrosine-specific phosphatases, which controls multiple cellular regulatory processes by removing phosphate ester groups from the tyrosine receptor. PTPN2 is generally expressed, but is most abundant in hematopoietic and placental cells (Mosinger, B. Jr et al., Proc Natl Acad Sci USA (1992) 89:499-503). In humans, PTPN2 expression is posttranscribedly controlled by the presence of two splice variants: a 45 kDa form containing a nuclear localization signal at the C-terminus upstream of the splice site and a 48 kDa typical form with a C-terminal ER retention motif (Tillmann U. et al., Mol Cell Biol (1994) 14:3030-3040). The 45 kDa isoform can be passively infused into the cytosol under certain cellular stress conditions. The two isoforms share the N-terminal phosphate-tyrosine phosphatase catalytic domain, and as a key negative regulator of the JAK-STAT pathway, PTPN2 directly regulates signal transduction via interleukin receptors. The PTPN2 catalytic domain shares 74% sequence homology with PTPN1 (also known as PTP1B) and shares similar enzyme kinetics (Romsicki Y. et al., Arch Biochem Biophys (2003) 414:40-50).

[0006] T-cell protein tyrosine phosphatase PTPN2 has been further identified as a key negative regulator of TCR signaling, thus highlighting the association between PTPN2 single nucleotide polymorphisms (SNPs) and autoimmune diseases (Wiede F et al., J Clin Invest. (2011);121(12):4758-4774). PTPN2 dephosphorylates and deactivates Src family kinases to regulate T-cell responses. PTPN2 deficiency has been shown to reduce the in vivo threshold for TCR-dependent CD8+ T-cell proliferation. Consistent with these findings, T-cell-specific PTPN2-deficient mice have been shown to exhibit widespread inflammation and autoimmunity. This autoimmunity is associated with increased serum levels of pro-inflammatory cytokines, antinuclear antibodies, T-cell infiltration in non-lymphocytic tissues, and liver disease. These data further indicate that PTPN2 is a key negative regulator of TCR signaling, setting the threshold for TCR-induced initial T-cell responses to prevent autoimmune and inflammatory conditions.

[0007] Besides PTPN2, which encodes T-cell PTP (TCPTP), serving as a susceptibility locus for autoimmune diseases, SNPs in PTPN2 have been associated with the development of type 1 diabetes, rheumatoid arthritis, and Crohn's disease. Furthermore, the type 1 diabetes-associated PTPN2 variant rs1893217 (C) is also associated with reduced PTPN2 expression in T cells (Florian Wiede, J Clin Invest. 2011;121(12):4758-4774).

[0008] The above findings suggest that inhibition of PTPN2 is a potential therapeutic strategy to improve the efficacy of cancer therapies associated with ICB resistance. Summary of the Invention

[0009] This invention relates to compounds, their pharmaceutically acceptable salts, pharmaceutical compositions thereof, and combinations thereof, which are effective inhibitors of protein tyrosine phosphatases, such as protein tyrosine phosphatase non-receptor type 2 (PTPN2) and / or protein tyrosine phosphatase non-receptor type 1 (PTPN1, also known as protein tyrosine phosphatase-1B (PTP1B)). The invention further provides a method for treating, preventing, or improving cancer, comprising administering an effective amount of the PTPN2 / PTPN1 inhibitor disclosed herein to an individual in need. In a preferred embodiment, the compound has a monocyclic core structure compared to compounds reported in the literature, wherein the compound contains a fused bicyclic core.

[0010] In some embodiments, this document discloses an inhibitor of a protein tyrosine phosphatase (e.g., PTPN2 and / or PTP1B) comprising a compound disclosed herein, such as a compound of formula (I). In other embodiments, this document discloses a method of treating a disease or condition (e.g., cancer, type 2 diabetes, obesity, metabolic disease, or any other disease, condition, or ailment) that responds advantageously to treatment with a PTPN2 or PTP1B inhibitor, comprising administering an effective amount of a compound disclosed herein, such as a compound of formula (I). These and other features of the invention will be set forth in an expanded form as the invention continues.

[0011] The first embodiment of the present invention provides at least one compound of formula (I): Formula (I) Among them, each occurrence is independent: R1 series are selected from the following groups: -H, alkyl, -OCH3, substituted alkyl, alkoxy, amine, secondary amine, tertiary amine, halogen, aryl, -CH2CH3, -CN, -OCH3, cyclopropyl, cyclopropoxy, cyclohexyl, -CF3, -OH, -Ph, -N(CH3)2, -NHCH3 and cycloalkyl; The R2 group is selected from the following groups: -H, alkyl, -CN, -OCH3, cycloalkyl, -CF3, -C(CH3)2R7, aryl, substituted alkyl, alkoxy, -CH(CH3)2, -C(CH3)3, -OCF3, -OH, and benzyloxy. R3 groups are selected from the following groups: -H, alkyl, -OCH3, substituted alkyl, amine, secondary amine, tertiary amine, -CHF2, halogen, -CN, -OCH3, -N(CH3)2, -OCHF2, alkoxy, -NHCH3, -OH, -CH2CH3, and α-lin-4-yl; The R4 series is selected from the following groups: -H, alkyl, -CH2CH3, -OCH3, -OH, and -CF3; R5 series are selected from the following groups: -H, cycloalkyl, alkyl and substituted alkyl.

[0012] Further, a compound selected from the following group of components is revealed: 5-[6-fluoro-4-[[(6-methoxy-2-pyridinyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one 5-(6-fluoro-4-(((5-methoxypyridin-2-yl)amino)methyl)-1H-indazol-7-yl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide 5-[6-fluoro-4-[[(4-methoxy-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one 5-[4-[[(4-cyclopropyl-2-pyridinyl)amino]methyl]-6-fluoro-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one 5-[6-fluoro-4-[[(4-methyl-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one 5-(4-(((4,6-dimethylpyridin-2-yl)amino)methyl)-6-fluoro-1H-indazol-7-yl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide 5-(6-fluoro-4-(((4-methoxy-5-methylpyridin-2-yl)amino)methyl)-1H-indazol-7-yl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide 6-[[6-fluoro-7-(1,1,4-trilateral-1,2,5-thiadiazolidin-2-yl)-1H-indazol-4-yl]methylamino]-4-methylpyridine-3-carboxynitrile 5-[6-fluoro-4-[[(4-methoxy-6-methyl-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one 5-[6-fluoro-4-[[(3-methyl-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one

[0013] In some embodiments, the compound of formula (I) is formulated into a pharmaceutically acceptable composition comprising the compound of formula (I) and a pharmaceutically acceptable carrier.

[0014] This document also discloses a method for treating cancer in patients of need, comprising administering to the patient an effective amount of a compound of formula (I) disclosed herein, and an additional therapeutic agent. In some embodiments, the additional therapeutic agent is an immunotherapeutic agent. For example, in some embodiments, the immunotherapeutic agent is an antibody.

[0015] This article also discloses a method for treating cancer in patients in need, which involves administering to the patient an effective amount of the compounds disclosed herein, such as compounds of formula (I).

[0016] This article further discloses a method for treating metabolic diseases in patients in need, comprising administering to the patient an effective amount of the compound disclosed herein, such as the compound of formula (I).

[0017] In some embodiments, the method includes treating cancer. In some embodiments, the cancer includes pancreatic cancer, breast cancer, multiple myeloma, melanoma, or cancer of secretory cells.

[0018] This document also discloses a composition for treating cancer in patients of need, wherein the composition comprises a compound disclosed herein (e.g., a compound of formula (I)) and an additional therapeutic agent. In some embodiments, the additional therapeutic agent is an immunotherapeutic agent. For example, in some embodiments, the immunotherapeutic agent is selected from the group consisting of anti-PD-1 antibodies and anti-PD-L1 antibodies.

[0019] This article further discloses a composition for treating metabolic diseases in patients in need, wherein the composition comprises compounds disclosed herein, such as compounds of formula (I). Implementation

[0020] Interactive reference for related applications

[0021] This application claims the benefit of U.S. Provisional Application No. 63 / 383,021, filed November 9, 2022, which is incorporated herein by reference in its entirety.

[0022] This invention relates to compounds, their pharmaceutically acceptable salts, pharmaceutical compositions thereof, and combinations thereof, which are effective inhibitors of protein tyrosine phosphatases, such as protein tyrosine phosphatase non-receptor type 2 (PTPN2) and / or protein tyrosine phosphatase non-receptor type 1 (PTPN1, also known as protein tyrosine phosphatase-1B (PTP1B)). The invention further provides a method for treating, preventing, or improving cancer, comprising administering an effective amount of the PTPN2 / PTPN1 inhibitor disclosed herein to an individual in need. In a preferred embodiment, the compound has a monocyclic core structure compared to compounds reported in the literature, wherein the compound contains a fused bicyclic core.

[0023] [definition] [] Chemical definition The following describes the definitions of specific functional groups and chemical terms in more detail. Chemical elements are identified according to the Periodic Table of the Elements, CAS version, Handbook of Chemistry and Physics, 75th edition, and specific functional groups are generally defined as described therein. Furthermore, the general principles of organic chemistry, as well as specific functional groups and reactivity, are described in the following: Thomas Sorrell, Organic Chemistry, University Science Books, Sausalito, 1999; Smith and March, March's Advanced Organic Chemistry, 5th edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; and Carruthers, Some Modern Methods of Organic Synthesis, 3rd edition, Cambridge University Press, Cambridge, 1987.

[0024] The abbreviations used in this article have their common meanings in the fields of chemistry and biology. The chemical structures and formulas described in this article are constructed according to the standard rules of chemical valence known in chemical techniques.

[0025] The compounds described herein may contain one or more asymmetric centers and therefore may exist in various isomeric forms, such as mirror-image isomers and / or non-mirror-image isomers. For example, the compounds described herein may be in the form of individual mirror-image isomers, non-mirror-image isomers, or geometric isomers, or mixtures of stereoisomers, including racemic mixtures and mixtures enriched with one or more stereoisomers. Isomers may be isolated from mixtures by methods known to those skilled in the art, including antipallic high-performance liquid chromatography (HPLC) and the formation and crystallization of antipallic salts; or preferred isomers may be prepared by asymmetric synthesis. See, for example, Jacques et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen et al., Tetrahedron 33:2725 (1977); Eliel, Stereochemistry of Carbon Compounds (McGraw-Hill, NY, 1962); and Wilen, Tables of Resolving Agents and Optical Resolutions, p. 268 (EL Eliel, ed., Univ. of Notre Dame Press, Notre Dame, IN 1972). This invention further covers the compounds described herein in their individual isomer forms, substantially free of other isomers, and alternatively in mixtures of various isomers.

[0026] In the compositions provided herein, the mirror-isomer pure compound may be present together with other active or inactive ingredients. For example, a pharmaceutical composition containing a mirror-isomer pure R-compound may contain, for example, about 90% excipients and about 10% mirror-isomer pure R-compound.

[0027] Given the following definitions, the features and advantages of the invention as described herein will be more readily understood by those skilled in the art. Certain features of the invention described in the context of individual embodiments may also be combined to form a single or extrapolated embodiment to include multiple embodiments. The illustrative or preferred embodiments identified herein are descriptive and not limiting.

[0028] Unless otherwise expressly stated herein, references in the singular may also include the plural. For example, "a" and "an" may refer to one, or one or more.

[0029] As used herein, the phrase "compound" refers to at least one compound. For example, a compound of formula (I) includes a compound of formula (I) and two or more compounds of formula (I).

[0030] Unless otherwise indicated, any heteroatom having an unsaturated valence is assumed to have hydrogen atoms sufficient to saturate the valence.

[0031] The definitions set forth herein take precedence over those set forth in any patent, patent application and / or patent application publication incorporated herein by reference.

[0032] The following lists the definitions of various terms used to describe the present invention. These definitions apply to the terms as they are used individually throughout the specification or as part of a larger group (unless otherwise limited in specific circumstances).

[0033] Throughout this specification, those familiar with the field can select groups and their substituents to provide stable moieties and compounds.

[0034] According to the conventions used in this technology, The bonds used in the structural formulas of this paper are used to describe the connection points between the parts or substituents and the core or main chain structure.

[0035] As used in this article, the terms "halogen" and "halogen" refer to F, Cl, Br, and I.

[0036] The term "cyano" refers to the group -CN.

[0037] The term "amine" refers to the group -NH2.

[0038] The term "side oxygen group" refers to the =O group.

[0039] As used herein, the term "alkyl" refers to a branched or straight-chain saturated aliphatic hydrocarbon group containing, for example, 1 to 12 carbon atoms, 1 to 6 carbon atoms, and 1 to 4 carbon atoms. Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (e.g., n-propyl and isopropyl), butyl (e.g., n-butyl, isobutyl, secondary butyl, and tertiary butyl), and pentyl (e.g., n-pentyl, isopentyl, and neopentyl), n-hexyl, 2-methylpentyl, 2-ethylbutyl, 3-methylpentyl, and 4-methylpentyl. When a number appears in subscript form after the symbol "C," the subscript more specifically defines the number of carbon atoms that a particular group may contain. For example, "C 1-6 alkyl" indicates a straight-chain or branched alkyl group having one to six carbon atoms.

[0040] As used herein, the term "fluoroalkyl" is intended to include branched and straight-chain saturated aliphatic hydrocarbon groups substituted with one or more fluorine atoms. For example, "C1-4 fluoroalkyl" is intended to include C1, C2, C3, and C4 alkyl groups substituted with one or more fluorine atoms. Representative examples of fluoroalkyl groups include, but are not limited to, -CF3 and -CH2CF3.

[0041] The term "cyanoalkyl" includes branched and straight-chain saturated alkyl groups substituted with one or more cyano groups. For example, "cyanoalkyl" includes -CH2CN, -CH2CH2CN, and C1-4 cyanoalkyl.

[0042] The term "aminoalkyl" includes branched and straight-chain saturated alkyl groups substituted with one or more amino groups. For example, "aminoalkyl" includes -CH2NH2, -CH2CH2NH2, and C1-4 aminoalkyl.

[0043] The term "hydroxyalkyl" includes branched and straight-chain saturated alkyl groups substituted with one or more hydroxyl groups. For example, "hydroxyalkyl" includes -CH2OH, -CH2CH2OH, and C1-4 hydroxyalkyl.

[0044] The term "hydroxy-fluoroalkyl" includes branched and straight-chain saturated alkyl groups substituted with one or more hydroxyl groups and one or more fluorine atoms. For example, "hydroxy-fluoroalkyl" includes -CHFCH2OH, -CH2CHFC(CH3)2OH and C1-4 hydroxy-fluoroalkyl.

[0045] As used herein, the terms "cycloalkyl," "carbocyclic," and "carbocyclic group" refer to groups derived from non-aromatic monocyclic or polycyclic hydrocarbon molecules by removing a hydrogen atom from a self-saturated carbon atom of a ring. Representative examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclopentyl, and cyclohexyl. When a number appears as a subscript after the symbol "C," the subscript more specifically defines the number of carbon atoms that a particular cycloalkyl group may contain. For example, "C 3-C 6 cycloalkyl" indicates a cycloalkyl group having three to six carbon atoms.

[0046] As used in this article, the term "heterocyclic" refers to organic compounds that have a cyclic structure consisting of both carbon atoms and non-carbon atoms, such as oxygen and nitrogen.

[0047] As used herein, the term "alkoxy" refers to an alkyl group, such as a methoxy group (-OCH 3), which is attached to a parent molecule via an oxygen atom. For example, "C 1-3 alkoxy" means an alkoxy group having one to three carbon atoms.

[0048] As used herein, the term "alkoxyalkyl" refers to an alkoxy group attached to an alkyl group via its oxygen atom, which is attached to a parent molecule moiety, such as methoxymethyl (˗CH 2OCH 3). For example, "C 2-4 alkoxyalkyl" means an alkoxyalkyl group having two to four carbon atoms, such as -CH 2OCH 3, -CH 2CH 2OCH 3, -CH 2OCH 2CH 3, and -CH 2CH 2OCH 2CH 3.

[0049] As used herein, the term "amine" or "amines" refers to a compound in which a nitrogen atom is directly bonded to several carbon atoms. Examples include derivatives of ammonia (-NH3) produced by the gradual substitution of three hydrogen atoms by a hydrocarbon group. Amines are classified as primary, secondary, or tertiary based on the number of carbons bonded to the nitrogen atom. For example, a primary amine has one carbon bonded to nitrogen (R-NH2), a secondary amine has two carbons bonded to both nitrogen and amine (R2-NH), and a tertiary amine has three carbons bonded to nitrogen (R3-N), where R is an alkyl group.

[0050] As used herein, "heteroaryl" refers to an aromatic heterocycle having 5 to 10 members, having at least one heteroatom selected from nitrogen, oxygen and sulfur, and containing at least one carbon atom, including monocyclic and bicyclic systems.

[0051] The phrase "medically acceptable" in this article means that such compounds, substances, compositions and / or dosage forms are suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic reactions or other problems or complications, within the bounds of reasonable medical judgment, and in proportion to a reasonable benefit / risk ratio.

[0052] The compound of formula (I) may be provided in amorphous or crystalline solid form. The compound of formula (I) in amorphous solid form may be provided by lyophilization.

[0053] It should be further understood that solvates (e.g., hydrates) of compounds of formula (I) are also within the scope of this invention. The term "solvate" means the physical association of a compound of formula (I) with one or more solvent molecules (organic or inorganic). This physical association includes hydrogen bonding. In some cases, such as when one or more solvent molecules are incorporated into the lattice of a crystalline solid, the solvate can be separated. "Solvate" encompasses both the solution phase and the separable solvate. Exemplary solvates include hydrates, ethanolates, methanolates, isopropanolates, acetonitrile solvates, and ethyl acetate solvates. The methods of fusion are known in this art.

[0054] Various forms of prodrugs are well known in this art and are described below: a) The Practice of Medicinal Chemistry, Camille G. Wermuth et al., Ch 31, (Academic Press, 1996); b) Design of Pro-drugs, edited by H. Bundgaard, (Elsevier, 1985); c) A Textbook of Drug Design and Development, edited by P. Krogsgaard-Larson and H. Bundgaard, Ch 5, pp. 113-191 (Harwood Academic Publishers, 1991); and d) Hydrolysis in Drug and Prodrug Metabolism, Bernard Testa and Joachim M. Mayer, (Wiley-VCH, 2003).

[0055] Furthermore, the compounds of formula (I) can be isolated and purified after their preparation to obtain a composition containing 99% by weight or more of the compounds of formula (I) ("substantially pure"), which is then used or formulated as described herein. Such "substantially pure" compounds of formula (I) are also covered herein as part of the present invention.

[0056] "Stable compound" and "stable structure" mean a compound that is sufficiently robust to withstand separation from the reaction mixture to a suitable purity and formulation into an effective therapeutic agent. This invention is intended to embody stable compounds.

[0057] Those skilled in the art will also understand that the compounds described and claimed herein as embodiments of the invention also exist in their "tautomeric form." As used herein, tautomers existing in tautomeric form refer to compounds that are structural isomers that can readily interconvert under rapid equilibrium. As used herein, the process of interconversion is referred to as "tautomerization."

[0058] For example, in the following embodiments, indazole tautomers can be represented as follows:

[0059] The revealed structure is easily convertible between left-handed and right-handed structural representations.

[0060] "Therapeutic effective amount" is intended to include the amount of a single compound of the invention, the amount of a combination of the claimed compounds, or the amount of a combination of a compound of the invention with other active ingredients that are effective as inhibitors or effective in treating or improving cancer.

[0061] As used herein, “treating” or “treatment” encompasses treating a disease state in mammals, particularly humans, and includes: (a) preventing the development of the disease state in mammals, especially when such mammals are predisposed to the disease state but have not yet been diagnosed with it; (b) suppressing the disease state, i.e., halting its development; and / or (c) alleviating the disease state, i.e., achieving the remission of the disease state.

[0062] The compounds of this invention are intended to include all isotopes of the atoms appearing in the compounds of this invention. Isotopes include atoms with the same number of atoms but different mass numbers. As a general example, but not a limitation, isotopes of hydrogen include deuterium (D) and tritium (T). Isotopes of carbon include 13C and 14C. The isotope-labeled compounds of this invention can generally be prepared by means of conventional techniques known to those skilled in the art or by methods similar to those described herein, using appropriately isotopically labeled reagents instead of the originally used unlabeled reagents. For example, methyl (-CH3) also includes deuterated methyl groups, such as -CD3.

[0063] The term "pharmaceutically acceptable salt" means including salts of active compounds prepared with relatively non-toxic acids or bases, depending on the specific substituents found on the compounds described herein. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds in their neutral form with a sufficient amount of the desired base in a solvent-free environment or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amine, magnesium, or similar salts.

[0064] As defined herein, the terms “inhibition,” “inhibit,” “inhibiting,” and the like in relation to protein-inhibitor (e.g., antagonist) interactions mean adversely affecting (e.g., reducing) the activity or function of a protein relative to its activity or function in the absence of an inhibitor. In some embodiments, inhibition refers to reducing disease or disease symptoms. In some embodiments, inhibition refers to reducing the activity of a signal transduction pathway or signaling pathway. Thus, inhibition includes at least partially, partially, or completely blocking stimulation, reducing, preventing, or delaying activation, or deactivating, desensitizing, or downregulating the activity or amount of signal transduction or enzymes or proteins. In some embodiments, inhibition refers to reducing the activity of protein tyrosine phosphatases, such as protein tyrosine phosphatase non-receptor type 2 (PTPN2) or protein tyrosine phosphatase non-receptor type 1 (PTP1B). Therefore, inhibition may include at least partially, partially or completely reducing stimulation; reducing or decreasing activation or inactivation; desensitizing or downregulating the amount of signal transduction or enzyme activity or protein tyrosine phosphatase, such as protein tyrosine phosphatase non-receptor type 2 (PTPN2) or protein tyrosine phosphatase non-receptor type 1 (PTP1B).

[0065] The term "patient" or "individual" in need refers to a living organism that suffers from or is susceptible to a disease or symptom that can be treated by administration of the compounds or pharmaceutical compositions provided herein. Non-limiting examples include humans, other mammals, bovine animals, rats, mice, dogs, monkeys, goats, sheep, dairy cows, deer, and other non-mammals. In some embodiments, the patient is a human. In some embodiments, the patient is a domestic animal. In some embodiments, the patient is a dog. In some embodiments, the patient is a parrot. In some embodiments, the patient is a livestock animal. In some embodiments, the patient is a mammal. In some embodiments, the patient is a cat. In some embodiments, the patient is a horse. In some embodiments, the patient is a cow. In some embodiments, the patient is a dog. In some embodiments, the patient is a feline. In some embodiments, the patient is an ape. In some embodiments, the patient is a monkey. In some embodiments, the patient is a mouse. In some embodiments, the patient is a laboratory animal. In some embodiments, the patient is a rat. In some embodiments, the patient is a hamster. In some embodiments, the patient is a laboratory animal. In some embodiments, the patient is a neonatal animal. In some embodiments, the patient is a neonatal human. In some embodiments, the patient is a neonatal mammal. In some embodiments, the patient is an aged animal. In some embodiments, the patient is an aged human. In some embodiments, the patient is an aged mammal. In some embodiments, the patient is an elderly patient.

[0066] "Disease," "symptom," or "condition" means a vital condition or health status of a patient or individual that can be treated with the compounds, pharmaceutical compositions, or methods provided herein. In some embodiments, the compounds and methods described herein comprise reducing or eliminating one or more symptoms of a disease, symptom, or condition, for example by administering a compound disclosed herein, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising a compound disclosed herein or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0067] As used herein, the term "signaling pathway" refers to a series of interactions between a cell and, where appropriate, extracellular components (e.g., proteins, nucleic acids, small molecules, ions, lipids) that transmit changes in one component to one or more other components, which in turn transmit changes to additional components, thereby propagating to other signaling pathway components, where appropriate.

[0068] "Pharmaceutically acceptable excipients" and "pharmaceutically acceptable carriers" refer to substances that facilitate the administration of an active agent to an individual, are absorbed by the individual, and can be included in the compositions of the present invention without causing significant adverse toxicological effects on the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, standard saline solutions, lactated Ringer's solution, standard sucrose, standard glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavorings, salt solutions (such as Ringer's solution), alcohols, oils, gelatin, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethyl cellulose, polyvinylpyrrolidone, and pigments, and the like. Such formulations may be sterilized and, as needed, mixed with adjuvants (such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, colorants, and / or aromatic substances and the like) that will not adversely react with the compounds of the present invention. Those skilled in this art will recognize that other pharmaceutical excipients are applicable to this invention.

[0069] The term "formulation" is intended to include the formulation of an active compound together with an encapsulating substance as a carrier to provide a capsule in which the active component, with or without another carrier, is surrounded by the carrier, thus associating the carrier with the capsule.

[0070] Similarly, this includes capsules and lozenges. Lozenges, powders, capsules, pills, capsules, and lozenges can be used as solid dosage forms suitable for oral administration.

[0071] As used herein, the term "administration" means administration to an individual via oral, suppository, local contact, intravenous, non-enteric, intraperitoneal, intramuscular, intralesional, intrathecal, intracranial, intranasal, or subcutaneous delivery, or via implantation of a sustained-release device, such as a micro-osmotic pump. Administration is carried out via any route, including non-enteric and mucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or percutaneous). Non-enteric administration includes, for example, intravenous, intramuscular, intraarterial, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial delivery. Other delivery modalities include, but are not limited to, the use of liposome formulations, intravenous infusion, and percutaneous patches. "Co-administration" means administration of the compound or composition described herein immediately before or immediately after administration of one or more additional therapies (e.g., anticancer agents, chemotherapy, or immunotherapy agents). The compound or composition described herein may be administered to a patient alone or co-administered. Co-doping and intent include the simultaneous or sequential dosing of compounds or compositions individually or in combination (more than one compound or agent). Therefore, formulations can also be combined with other active substances when needed (e.g., to reduce metabolic degradation).

[0072] The pharmaceutical compositions described herein can be prepared by any method known in pharmacological techniques. Generally, such preparation methods include the following steps: associating the disclosed compound ("active ingredient") with a carrier and / or one or more other adjuncts, and then, as needed and / or required, shaping and / or packaging the product into a desired single-dose or multiple-dose units. Pharmaceutical compositions can be prepared, packaged, and / or sold in batches, as single-unit doses, and / or as multiple single-unit doses. As used herein, a "unit dose" is an individual amount of a pharmaceutical composition containing a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dose of the active ingredient administered to an individual and / or an appropriate fraction of such dose, such as one-half or one-third of such dose.

[0073] [Treatment methods] [] The present invention is characterized by compounds, compositions, and methods comprising compounds disclosed herein, such as compounds of formula (I). In some embodiments, the compounds, compositions, and methods disclosed herein are used to prevent or treat diseases, conditions, or symptoms. Examples of diseases, conditions, or symptoms include, but are not limited to, cancer, type 2 diabetes, metabolic syndrome, obesity, or metabolic diseases.

[0074] [cancer] [] In some embodiments, the compounds disclosed herein (e.g., compounds of formula (I)) are used to treat cancer. As used herein, "cancer" refers to human cancers and carcinomas, sarcomas, adenocarcinomas (e.g., papillary adenocarcinoma), lymphomas, leukemias, melanomas, etc., including solid carcinomas and lymphomas, kidney cancer, breast cancer, lung cancer, bladder cancer, colon cancer, ovarian cancer, prostate cancer, pancreatic cancer, stomach cancer, brain cancer, head and neck cancer, skin cancer, uterine cancer, testicular cancer, gliomas, esophageal cancer, liver cancer including hepatocarcinoma, lymphomas including B-cell acute lymphoblastic lymphoma, non-Hodgkin's lymphomas (e.g., Burkitt's lymphomas, small cell lymphomas, and large cell lymphomas), Hodgkin's lymphoma, leukemias (including AML, ALL, and CML), and / or multiple myeloma. In some other contexts, "cancer" refers to lung cancer, breast cancer, ovarian cancer, epithelial ovarian cancer, leukemia, lymphoma, melanoma, pancreatic cancer, sarcoma, bladder cancer, bone cancer, bile duct cancer, adrenal cancer, salivary gland cancer, bronchial cancer, oral cancer, oral cavity or pharyngeal cancer, laryngeal cancer, kidney cancer, gynecological cancer, brain cancer, central nervous system cancer, peripheral nervous system cancer, blood tissue cancer, small intestine or appendix cancer, cervical cancer, colon cancer, esophageal cancer, stomach cancer, liver cancer, head and neck cancer, kidney cancer, myeloma, thyroid cancer, prostate cancer, metastatic cancer, or tumor.

[0075] Exemplary cancers that can be treated with the compounds, pharmaceutical compositions, or methods provided herein include lymphoma, B-cell lymphoma, heavy chain disease, alpha chain disease, gamma chain disease, μ chain disease, Waldenstrom's macroglobulinemia, benign monoclonal gamma globulinopathy, sarcoma, bladder cancer, bone cancer, brain tumor, cervical cancer, colon cancer, esophageal cancer, gastric cancer, head and neck cancer, kidney cancer, myeloma, thyroid cancer, leukemia, prostate cancer, and breast cancer (e.g., ER-positive breast cancer, ER-negative breast cancer, chemotherapy-resistant breast cancer, herceptin-resistant breast cancer, HER2-positive breast cancer, doxorubicin-resistant breast cancer, tamoxifen...). Oxifen-resistant breast cancer, ductal carcinoma, lobular carcinoma, primary breast cancer, metastatic breast cancer, ovarian cancer, pancreatic cancer, liver cancer (e.g., hepatocellular carcinoma), lung cancer (e.g., non-small cell lung cancer, squamous cell lung cancer, adenocarcinoma, large cell lung cancer, small cell lung cancer, carcinoid, sarcoma), glioblastoma multiforme, acoustic neuroma, retinoblastoma, astrocytoma, craniopharyngioma, hemangioblastoma, pineal tumor, ependymoma, oligodendritic glioma, meningioma, glioma, or melanoma. Additional examples include thyroid cancer, endocrine system cancers, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, liver cancer, kidney cancer, lung cancer, non-small cell lung cancer, melanoma, mesothelioma, ovarian cancer, sarcoma, stomach cancer, uterine cancer or neuroblastoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, immune-mediated amyloidosis, ovarian cancer, rhabdomyosarcoma, and so on. Primary thrombocytosis, primary macroglobulinemia, primary brain tumor, cancer, malignant pancreatic islet tumor, malignant carcinoid tumor, bladder cancer, precancerous skin lesions, testicular cancer, lymphoma, thyroid cancer, neuroblastoma, esophageal cancer, urogenital tract cancer, malignant hypercalcemia, endometrial cancer, adrenocortical cancer, endocrine or exocrine pancreatic lesions, medullary thyroid carcinoma, medullary thyroid carcinoma, melanoma, colorectal cancer, papillary thyroid carcinoma, and hepatocellular carcinoma.

[0076] The first embodiment of the present invention provides at least one compound of formula (I): A compound having the following structure: Among them, each occurrence is independent: R1 series are selected from the following groups: -H, alkyl, -OCH3, substituted alkyl, alkoxy, amine, secondary amine, tertiary amine, halogen, aryl, -CH2CH3, -CN, -OCH3, cyclopropyl, cyclopropoxy, cyclohexyl, -CF3, -OH, -Ph, -N(CH3)2, -NHCH3 and cycloalkyl; The R2 group is selected from the following groups: -H, alkyl, -CN, -OCH3, cycloalkyl, -CF3, -C(CH3)2R7, aryl, substituted alkyl, alkoxy, -CH(CH3)2, -C(CH3)3, -OCF3, -OH, and benzyloxy. R3 groups are selected from the following groups: -H, alkyl, -OCH3, substituted alkyl, amine, secondary amine, tertiary amine, -CHF2, halogen, -CN, -OCH3, -N(CH3)2, -OCHF2, alkoxy, -NHCH3, -OH, -CH2CH3, and α-lin-4-yl; The R4 series is selected from the following groups: -H, alkyl, -CH2CH3, -OCH3, -OH, and -CF3; R5 series are selected from the following groups: -H, cycloalkyl, alkyl and substituted alkyl.

[0077] In one embodiment of the compound of formula I: R1 is selected from the following groups: -H, -CH3, -OCH3, and cyclopropyl; R2 series are selected from the following groups: -H, -CH3, -CN, and -OCH3; R3 series are selected from the following groups: -H, -CH3, and -OCH3; R4 series is selected from the following groups: -H and -CH3.

[0078] In another embodiment of the compound of formula I: R1 is cyclopropyl; R2 is -H; R 3 is -H; R4 is -H; R 5 is -H.

[0079] In one embodiment of the compound of formula I: R1 is -OCH 3; R2 is an alkyl group; R 3 is -H; R4 is -H; R 5 is -H.

[0080] In another embodiment of the compound of formula I: R1 is -OCH 3; R2 is -H; R 3 is -H; R4 is -H; R 5 is -H.

[0081] In one embodiment of the compound of formula I: R1 is -OCH 3; R2 is -H; R3 is an alkyl group; R4 is -H; R 5 is -H.

[0082] In one embodiment of the compound of formula I: R1 is -H; R2 is -OCH3; R 3 is -H; R4 is -H; R 5 is -H.

[0083] In another embodiment of the compound of formula I: R1 is an alkyl group; R2 is -CN; R 3 is -H; R4 is -H; R 5 is -H.

[0084] In one embodiment of the compound of formula I: R1 is -H; R2 is -H; R 3 is -H; R4 is an alkyl group; R 5 is -H.

[0085] In another embodiment of the compound of formula I: R1 is -H; R2 is -H; R 3 is -OCH 3; R4 is -H; R 5 is -H.

[0086] In one embodiment of the compound of formula I, the compound is selected from the group consisting of: 5-[6-fluoro-4-[[(6-methoxy-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one; 5-(6-fluoro-4-(((5-methoxypyridin-2-yl)amino)methyl)-1H-indazol-7-yl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide; 5-[6-fluoro-4-[[(4-methoxy-2-pyridinyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one; 5-[4-[[(4-cyclopropyl-2-pyridinyl)amino]methyl]-6-fluoro-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one; 5-[6-fluoro-4-[[(4-methyl-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideoxy-1,2,5-thiadiazolidin-3-one; 5-(4-(((4,6-dimethylpyridin-2-yl)amino)methyl)-6-fluoro-1H-indazol-7-yl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide; 5-(6-fluoro-4-(((4-methoxy-5-methylpyridin-2-yl)amino)methyl)-1H-indazol-7-yl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide; 6-[[6-fluoro-7-(1,1,4-trilateral-oxy-1,2,5-thiadiazolidin-2-yl)-1H-indazol-4-yl]methylamino]-4-methylpyridine-3-carboxynitrile; 5-[6-fluoro-4-[[(4-methoxy-6-methyl-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one; 5-[6-fluoro-4-[[(3-methyl-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideoxy-1,2,5-thiadiazolidin-3-one; Or a medically acceptable salt.

[0087] In another embodiment, the present invention comprises a pharmaceutical composition comprising a compound of formula (I) of technical solution 1 or a pharmaceutically acceptable salt thereof and at least one pharmaceutically acceptable carrier.

[0088] In one embodiment, the present invention includes a method for treating cancer, comprising administering to the patient a therapeutically effective amount of a compound of formula (I) of technical solution 1 or a pharmaceutically acceptable salt thereof, wherein the cancer / disease is selected from: human cancer, carcinoma, sarcoma, adenocarcinoma, papillary adenocarcinoma, lymphoma, leukemia, melanoma, solid lymphoma, kidney cancer, breast cancer, lung cancer, bladder cancer, colon cancer, ovarian cancer, prostate cancer, pancreatic cancer, stomach cancer, brain cancer, head and neck cancer, skin cancer, uterine cancer, testicular cancer, glioma, esophageal cancer, liver cancer including liver cancer, lymphoma, including B acute lymphoblastic lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, small lymphoma, Hodgkin's lymphoma, leukemia, and multiple myeloma.

[0089] In another embodiment, the present invention includes a method of treating cancer in a patient in need, comprising administering to the patient an effective amount of a compound of formula I and an additional therapeutic agent.

[0090] In one embodiment, the additional therapeutic agent is an immunotherapy agent.

[0091] In another embodiment, the immunotherapy agent is selected from the group consisting of: anti-PD-1 antibody, anti-PD-L1 antibody, and anti-CTLA-4 antibody.

[0092] In one embodiment, a method of treating cancer in a patient in need comprises administering to the patient an effective amount of a medically acceptable composition of a compound of formula I.

[0093] In another embodiment, the method of treating cancer is selected from radiation, surgery, chemotherapy or administration of biological drugs.

[0094] In one embodiment, the method of treating cancer is to administer a biological drug, which is a drug that stimulates the immune system.

[0095] In another embodiment, a method of treating cancer includes administering to an individual an inhibitor of DGKα and / or DGKζ, an antagonist of the PD1 / PD-L1 axis, and an antagonist of CTLA4.

[0096] These embodiments are not intended to limit the scope of the invention.

[0097] [Synthesis Method] [] The compounds of this invention can be prepared by the methods and examples presented below, as well as by methods known to those skilled in the art. In each of the examples below, unless otherwise specified, the R group is as defined above for each formula. Optimal reaction conditions and reaction times may vary depending on the reactants used. Unless otherwise stated, solvents, temperatures, pressures, and other reaction conditions can be readily selected by those skilled in the art.

[0098] Intermediates used in the following synthesis are commercially available or readily prepared by methods known to those skilled in the art. Reaction progress can be monitored by known methods, such as thin-layer chromatography (TLC) or high-performance liquid chromatography-mass spectrometry (HPLC-MS). Intermediates and products can be purified by methods known in this art, including column chromatography, HPLC, preparative TLC, or preparative HPLC.

[0099] Preparation Examples Preparation of synthetic intermediate (Int-2) [preparation] [7-(1,1-] [Dioxane] [-4-] [Side group] [-1,2,5-] [Thiadiazole] [-2-] [base] [)-6-] [fluorine] [-2-(4-)] [Methoxybenzyl] [)-2H-] [Indazole] [-4-] [formaldehyde] [(Int-2)] [,] [As per the process] [1] [As shown in the image.] [process] [1] [:] [] []

[0100] [step] [1] [:synthesis] [4-] [bromine] [-6-] [fluorine] [-7-] [Nitro] [-1H-] [Indazole] [(1-2)] []

[0101] At 0 °C, potassium nitrate (4.98 g, 49.30 mmol) dissolved in concentrated sulfuric acid (100 mL) was added dropwise to a round-bottom flask containing 10.00 g (46.95 mmol) of 4-bromo-6-fluoro-1H-indazole and 100 mL of sulfuric acid. The mixture was stirred overnight at room temperature. After the reaction was complete as monitored by LCMS, the mixture was poured into 1.0 L of ice water, resulting in a pale yellow precipitate. The mixture was filtered. The solid was washed with water and dried to give the desired product. The crude product was purified by silica gel column chromatography (5% to 10% ethyl acetate / petroleum ether) to give 4-bromo-6-fluoro-7-nitro-1H-indazole (9.33 g, 35.88 mmol, 76.42% yield) as a yellow solid. ¹H NMR (400 MHz, DMSO-d⁶) δ 14.23 (s, ¹H), 8.33 (d, J = 1.7 Hz, ¹H), 7.74 (dd, J = 11.5, 1.9 Hz, ¹H). Note: The desired isomer is identified by 2D NMR because there is no correlation between the aromatic proton and the NH group of the pyrazole moiety.

[0102] Step 2: Synthesis of 4-bromo-6-fluoro-2-(4-methoxybenzyl)-7-nitro-2H-indazole (1-3)

[0103] At room temperature, 2,2,2-trichloroacetylimine-4-methoxybenzyl ester (12.68 g, 44.98 mmol) and TsOH (1.21 g, 7.02 mmol) were added to a stirred mixture of 4-bromo-6-fluoro-7-nitro-1H-indazole (9.33 g, 35.88 mmol) in DCM (400 mL). The resulting mixture was stirred overnight at room temperature. LCMS showed complete depletion of the starting material. The solution was diluted with DCM and washed with saturated sodium bicarbonate. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography and dissolved in DCM to give 4-bromo-6-fluoro-2-[(4-methoxyphenyl)methyl]-7-nitro-indazole (12.22 g, 32.15 mmol, 89.60% yield) as a yellow solid. MS: m / z: C 15H 11BrFN 3O 3[M+H] + The calculated value is 380, and the experimental value is 380. []

[0104] [step] [3] [:synthesis] [4-] [bromine] [-6-] [fluorine] [-2-(4-)] [Methoxybenzyl] [)-2H-] [Indazole] [-7-] [amine] [(1-4)] []

[0105] Fe (17.77 g, 318.26 mmol) and NH₄Cl (17.22 g, 321.91 mmol) were added to a stirred mixture of 4-bromo-6-fluoro-2-[(4-methoxyphenyl)methyl]-7-nitro-indazole (12.22 g, 32.15 mmol) in ethanol (200 mL) and water (20 mL) at room temperature. The resulting mixture was stirred at 80 °C under a nitrogen atmosphere for 2 h. The reaction was complete as indicated by LCMS. The reaction mixture was filtered. The filtrate was concentrated under reduced pressure to remove ethanol and diluted with water (200 mL). The solution was extracted with ethyl acetate (3 × 200 mL). The combined organic layers were washed with brine (200 mL), dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silica gel column chromatography (dichloromethane containing 6% ethyl acetate) to give 4-bromo-6-fluoro-2-[(4-methoxyphenyl)methyl]indazole-7-amine (10 g, 28.56 mmol, 88.82% yield) as a pink solid. MS: m / z: C 15H 13BrFN 3O [M+H]+ calculated value 350, experimental value 350.

[0106] [step] [4] [:] [synthesis] [(4-)] [bromine] [-6-] [fluorine] [-2-(4-)] [Methoxybenzyl] [)-2H-] [Indazole] [-7-] [base] [)] Ethyl glycine [(1-5)]

[0107] At 0 °C, TMSCl (6.8 mL, 53.54 mmol) was added to a stirred solution of 4-bromo-6-fluoro-2-[(4-methoxyphenyl)methyl]indazole-7-amine (7.5 g, 21.42 mmol) and 50% 2-sidektoethyl ethyl / toluene (6.58 g, 32.13 mmol) in DCM (100 mL). The mixture was stirred at ambient temperature for 40 min. At 0 °C, a solution of NBH 3CN (3.37 g, 53.54 mmol) in DCM (20 mL) was slowly added to the above mixture. The resulting mixture was stirred at ambient temperature for 3 h until the starting material was completely depleted. The mixture was quenched by adding saturated NH 4Cl (200 mL) at 0 °C. The solution was extracted twice with ethyl acetate. The combined organic phases were washed three times with brine. The organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated. The residue was purified by silicone column chromatography (20% to 30% ethyl acetate / petroleum ether) to give ethyl 2-[[4-bromo-6-fluoro-2-[(4-methoxyphenyl)methyl]indazol-7-yl]amino]ethyl acetate (5.5 g, 12.60 mmol, 58.86% yield) as a pale yellow oil. MS: m / z: C 19H 19BrFN 3O 3[M+H] + calculated value 436, experimental value 436.

[0108] [step] [5] [:] [synthesis] [N-(4-)] [bromine] [-6-] [fluorine] [-2-(4-)] [Methoxybenzyl] [)-2H-] [Indazole] [-7-] [base] [)-N-] [Ethyl amine sulfonylurea glycine] [(1-6)]

[0109] At 0 °C, a solution of aminesulfonyl chloride (9.47 g, 81.94 mmol) in DMA (15 mL) was added to a stirred solution of ethyl 2-[[4-bromo-6-fluoro-2-[(4-methoxyphenyl)methyl]indazol-7-yl]amino]ethyl acetate (5.5 g, 12.61 mmol) in DMA (50 mL). The reaction mixture was stirred overnight at ambient temperature. LCMS showed complete depletion of the starting material. The mixture was diluted with ethyl acetate and washed seven times with brine until the DMA was completely removed. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give 6.7 g of a light brown semi-solid product. The crude product was used directly in the next step. MS: m / z: C 19H 20BrFN 4O 5S [M+H]+ calculated value 515, experimental value 515.

[0110] [step] [6] [:] [synthesis] [5-(4-)] [bromine] [-6-] [fluorine] [-2-(4-)] [Methoxybenzyl] [)-2H-] [Indazole] [-7-] [base] [)-1,2,5-] [Thiadiazole] [-3-] [ketone] [1,1-] [Dioxide] [(Int-1)]

[0111] At 0 °C, MeOH (14.03 g, 78.01 mmol) containing 30% NaOMe was added to a stirred solution of ethyl 2-[[4-bromo-6-fluoro-2-[(4-methoxyphenyl)methyl]indazol-7-yl]-aminesulfonyl-amino]ethyl acetate (6.7 g, 13 mmol) in methanol (60 mL). The reaction mixture was stirred at ambient temperature for 2 h. After the reaction was completed as monitored by LCMS, the mixture was diluted with ethyl acetate and concentrated. The resulting suspension was dissolved in water (500 mL), diluted with ethyl acetate, acidified to pH 3 with 1 N HCl solution, and extracted three times with ethyl acetate. The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under vacuum. The residue was purified by reverse-phase column chromatography to give a pale yellow solid, 5-[4-bromo-6-fluoro-2-[(4-methoxyphenyl)methyl]indazol-7-yl]-1,1-dioxy-1,2,5-thiadiazolidin-3-one (4 g, 8.5237 mmol, 65.56% yield). MS: m / z: C 17H 14BrFN 4O 4S [M+H]+, calculated value 469, experimental value 469.

[0112] [step] [7] [:] [synthesis] [5-(6-)] [fluorine] [-2-(4-)] [Methoxybenzyl] [)-4-] [Vinyl] [-2H-] [Indazole] [-7-] [base] [)-1,2,5-] [Thiadiazole] [-3-] [ketone] [1,1-] [Dioxide] [(1-7)]

[0113] Pd₂(dba)₃ (0.39 g, 0.43 mmol) and P(t-Bu)₃HBF₄ (0.41 g, 0.85 mmol) were added to a stirred solution of 5-[4-bromo-6-fluoro-2-[(4-methoxyphenyl)methyl]indazol-7-yl]-1,1-dioxy-1,2,5-thiadiazolidine-3-one (2 g, 4.26 mmol) and tributyl(vinyl)stanane (4.05 g, 12.79 mmol) in DMA (20 mL). The resulting mixture was evacuated and backfilled three times with N₂. The mixture was then stirred overnight at 100 °C. LC-MS showed complete depletion of the starting material. The reaction mixture was purified by reverse-phase column chromatography (containing 0.05% NH₄HCO₃ water and MeCN) to give a yellow solid of 5-[6-fluoro-2-[(4-methoxyphenyl)methyl]-4-vinyl-indazol-7-yl]-1,1-dioxy-1,2,5-thiadiazolidine-3-one (1.1 g, 2.64 mmol, 61.98% yield). MS: m / z: C₁₉H₁₇FN₄O₄S [M+H]⁺, calculated value 417, experimental value 417.

[0114] [step] [8] [:] [synthesis] [7-(1,1-] [Dioxane] [-4-] [Side group] [-1,2,5-] [Thiadiazole] [-2-] [base] [)-6-] [fluorine] [-2-(4-)] [Methoxybenzyl] [)-2H-] [Indazole] [-4-] [formaldehyde] [(Int-2)]

[0115] At room temperature, K₂O₅₄ (0.09 g, 0.26 mmol) was added to a stirred solution of 5-[6-fluoro-2-[(4-methoxyphenyl)methyl]-4-vinyl-indazol-7-yl]-1,1-dioxy-1,2,5-thiadiazolidine-3-one (1.1 g, 2.64 mmol), NMO (0.62 g, 5.28 mmol), and citric acid (1.11 g, 5.28 mmol) in a mixture of tert-butanol (10 mL) and water (10 mL). The resulting mixture was stirred at room temperature for 2 h. LCMS showed complete conversion of the starting material to the intermediate. Subsequently, NaIO₄ (1.69 g, 7.92 mmol) was added to the mixture in portions at 0 °C. The resulting mixture was stirred at room temperature for 2 h. LCMS showed completion of the reaction. The reaction mixture was diluted with water (50 mL) and extracted seven times with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated. The resulting residue was purified by reverse-phase column chromatography (containing 0.05% NH₄HCO₃ water and MeCN) to give 6-fluoro-2-[(4-methoxyphenyl)methyl]-7-(1,1,4-trisyloxy-1,2,5-thiadiazolin-2-yl)indazole-4-carboxaldehyde (600 mg, 1.43 mmol, 54.28% yield) as a yellow solid. MS: m / z: C₁₈H₁₅FN₄O₅S [M+H]⁺, calculated value 419, experimental value 419.

[0116] [Example] [1] [:]5-[6-fluoro-4-[[(6-methoxy-2-pyridinyl)amino]methyl]-1H-indazol-7-yl]-1,1-dioxy-1,2,5-thiadiazolidin-3-one

[0117] [process] [2] [:] []

[0118] [step] [1] [:] [synthesis] [5-(6-)] [fluorine] [-2-(4-)] [Methoxybenzyl] [)-4-(((6-] [Methoxypyridine] [-2-] [base] [)] [Amine] [)] [methyl] [)-2H-] [Indazole] [-7-] [base] [)-1,2,5-] [Thiadiazole] [-3-] [ketone] [1,1-] [Dioxide] [(2-1)]

[0119] At 0°C, 6-fluoro-2-[(4-methoxyphenyl)methyl]-7-(1,1,4-trisyloxy-1,2,5-thiadiazolidin-2-yl)indazole-4-carboxaldehyde ( [Int-2] (60 mg, 0.14 mmol) and 6-methoxypyridine-2-amine (21.36 mg, 0.17 mmol) were added to a stirred solution in DCM (5 mL) along with trimethylsilane trifluoromethanesulfonate (63.89 mg, 0.28 mmol). The mixture was stirred at room temperature for 1 h. NaBH(OAc)3 (27.53 mg, 0.29 mmol) was added to the mixture at 0 °C. The reaction mixture was stirred again at room temperature for 2 hours. After the reaction was monitored by LCMS, the mixture was diluted with DCM (10 mL) and concentrated under vacuum. The residue was purified by reverse-phase column chromatography (containing 0.05% NH₄HCO₃ water and MeCN) to give a pale yellow solid of 5-[6-fluoro-1-[(4-methoxyphenyl)methyl]-4-[[(6-methoxy-2-pyridyl)amino]methyl]indazol-7-yl]-1,1-dioxy-1,2,5-thiadiazolin-3-one (50 mg, 0.095 mmol, 66.21% yield). MS: m / z: C₂₄H₂₃FN₆O₅S [M+H]⁺ calculated value was 527; experimental value was 527. []

[0120] [step] [2] [:] [synthesis] [5-[6-] [fluorine] [-4-[[(6-)] [Methoxy] [-2-] [Pyridyl] [)] [Amine] []] [methyl] []-1H-] [Indazole] [-7-] [base] []-1,1-] [Dilateral oxygen group] [-1,2,5-] [Thiadiazole] [-3-] [ketone] [(2-2)]

[0121] At room temperature, TFA (2 mL) was added to a solution of 5-[6-fluoro-1-[(4-methoxyphenyl)methyl]-4-[[(6-methoxy-2-pyridyl)amino]methyl]indazol-7-yl]-1,1-dioxy-1,2,5-thiadiazolidin-3-one (50 mg, 0.09 mmol) in DCE (2 mL), and the mixture was stirred at 60 °C for 4 h. After the reaction was completed, the mixture was concentrated by LCMS monitoring. The residue was purified by reverse-phase column chromatography (0.05% NH₄HCO₃ in water and MeCN) and further purified by preparative HPLC to give a white solid 5-[6-fluoro-4-[[(6-methoxy-2-pyridinyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolin-3-one (14.5 mg, 0.03 mmol, 37.08% yield). MS: m / z: C₁₆H₁₅FN₆O₄S [M+H]⁺, calculated value 407; experimental value 407. 1H NMR (400 MHz, DMSO- d 6) δ13.28 (s, 1H), 8.33 (d, J = 3.8 Hz, 1H), 7.58-7.13 (m, 2H), 7.01 (d, J = 11.0 Hz, 1H), 6.09 (dd, J = 7.9, 3.9 Hz, 1H), 5.90 (dd, J = 7.8, 3.9 Hz, 1H), 4.77 (s, 2H), 4.45 (d, J = 3.8 Hz, 2H), 3.66 (d, J = 3.0 Hz, 3H).

[0122] Preparative HPLC purification conditions: Column: SunFire preparative C18 OBD column, 19*150 mm, 5 μm; Mobile phase A: water (0.1% FA), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: 40% B to 60% B, 60% B over 6.5 min; Wavelength: 254 / 210 nm.

[0123] [Example] [2] [:]5-(6-fluoro-4-(((5-methoxypyridin-2-yl)amino)methyl)-1H-indazol-7-yl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide

[0124] The title compound, a white solid, was prepared in 11.53% overall yield using 5-methoxypyridine-2-amine from step 1, according to Example 1. MS: m / z: C 16H 15FN 6O 4S [M+H]+ calculated value 407; experimental value 407. 1H NMR (400 MHz, DMSO-d 6) δ 13.26 (s, 1H), 8.59 (s, 1H), 8.26 (s, 1H), 7.70 (dd, J = 9.7, 2.8 Hz, 1H), 7.54 (d, J = 2.9 Hz, 1H), 7.13–6.98 (m, 2H), 4.85 (s, 2H), 4.17 (s, 2H), 3.77 (s, 3H).

[0125] Preparative HPLC purification conditions: Column: HALO C18, 3.0*30 mm, 2.0 μM; Mobile phase A: Water / 0.05% TFA, Mobile phase B: ACN / 0.05% TFA; Flow rate: 1.5000 L / min; Gradient: 5% B to 40% B in 1.69 min, 40% B to 95% B in 0.60 min, and maintained at 95% B in 0.5 min; Wavelength: 254 nm.

[0126] [Example] [3]:5-[6-fluoro-4-[[(4-methoxy-2-pyridinyl)amino]methyl]-1H-indazol-7-yl]-1,1-dioxy-1,2,5-thiadiazolidin-3-one

[0127] The title compound, a white solid, was prepared in 21.62% overall yield using 4-methoxypyridine-2-amine from step 1, according to Example 1. MS: m / z: C 16H 15FN 6O 4S [M+H]+ calculated value 407; experimental value 407. 1H NMR (400 MHz, DMSO- d 6) b13.25 (s, 1H), 12.79 (s, 1H), 8.75 (s, 1H), 8.26 (s, 1H), 7.87 (d, J = 7.2 Hz, 1H), 7.06 (d, J = 11.5 Hz, 1H), 6.58 (dd, J = 7.2, 2.4 Hz, 1H), 6.48 (d, J = 2.4 Hz, 1H), 4.89 (d, J = 5.6 Hz, 2H), 4.12 (s, 2H), 3.90 (s, 3H).

[0128] Preparative HPLC purification conditions: Column: SunFire preparative C18 OBD column, 19*150 mm, 5 μm; Mobile phase A: water (0.05% TFA), Mobile phase B: ACN; Flow rate: 25 mL / min; Gradient: 20% B to 40% B, 40% B over 5.3 min; Wavelength: 254 / 210 nm.

[0129] [Example] [4]:5-[4-[[(4-cyclopropyl-2-pyridinyl)amino]methyl]-6-fluoro-1H-indazol-7-yl]-1,1-disidel-1,2,5-thiadiazolidin-3-one

[0130] The title compound, a white solid, was prepared in 7.78% overall yield using 4-cyclopropylpyridine-2-amine from step 1, according to Example 1. MS: m / z: C 18H 17FN 6O 3S [M+H]+, calculated value 417; experimental value 417. 1H NMR (400 MHz, DMSO- d 6+D2O) δ8.28 (d, J = 5.0 Hz, 1H), 7.77-7.67 (m, 1H), 7.07 (dd, J = 11.5, 4.2 Hz, 1H), 6.77 (d, J = 11.4 Hz, 1H), 6.54 (t, J = 6.8 Hz, 1H), 4.88 (d, J = 5.7 Hz, 2H), 4.15 (s, 2H), 2.10-1.90 (m, 1H), 1.30-1.10 (m, 2H), 1.00-0.70 (m, 2H).

[0131] Preparative HPLC purification conditions: Column: SunFire preparative C18 OBD column, 19*150 mm, 5 μm; Mobile phase A: water (0.1% FA), Mobile phase B: ACN; Flow rate: 20 mL / min; Gradient: 10% B to 25% B, 25% B over 6 min; Wavelength: 210 / 254 nm.

[0132] [Example] [5] [:]5-[6-fluoro-4-[[(4-methyl-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disidel-1,2,5-thiadiazolidine-3-one [] []

[0133] The title compound, a white solid, was prepared in 10.71% overall yield using 4-methylpyridin-2-amine from step 1, according to Example 1. MS: m / z: C 16H 15FN 6O 3S [M+H]+ calculated value 391; experimental value 391. ¹H NMR (400 MHz, DMSO-d 6) δ 13.28 (s, 1H), 8.99 (s, 1H), 8.26 (s, 1H), 7.87 (d, J = 6.5 Hz, 1H), 7.20–6.89 (m, 2H), 6.80 (d, J = 6.5 Hz, 1H), 4.88 (d, J = 5.5 Hz, 2H), 4.14 (s, 2H), 2.35 (s, 3H). []

[0134] Preparative HPLC purification conditions: Column: SunFire preparative C18 OBD column, 19*150 mm, 5 μm; Mobile phase A: water (0.1% FA), Mobile phase B: ACN; Flow rate: 25 mL / min; Gradient: 50% B to 70% B, 70% B over 5.3 min; Wavelength: 210 / 254 nm. []

[0135] [Example] [6] [:]5-(4-(((4,6-dimethylpyridin-2-yl)amino)methyl)-6-fluoro-1H-indazol-7-yl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide [] []

[0136] The title compound, a white solid, was prepared in 13.75% overall yield using 4,6-dimethylpyridin-2-amine from step 1, according to Example 1. MS: m / z: C 17H 18FN 6O 3S [M+H]+ calculated value 405; experimental value 405. 1H NMR (500 MHz, DMSO- d 6) δ13.28 (s, 1H), 8.99 (s, 1H), 8.26 (s, 1H), 7.21 (s, 1H), 7.11 (s, 1H), 7.07-6.98 (m, 2H), 6.74 (br s, 1H), 6.67-6.55 (m, 1H), 4.89 (br d, J=5.2 Hz, 2H), 4.11 (br s, 2H), 2.40 (s, 3H), 2.27 (s, 3H). []

[0137] Preparative HPLC purification conditions: XBridge C18 column, 19*200 mm, 5 μM; mobile phase A: ACN / H2O with 10 mM AA (5:95); mobile phase B: ACN / H2O with 10 mM AA (95:5); flow rate: 20 ml / min; gradient: 0% B to 40% B, 40% B over 20 min; wavelength: 220 nm. []

[0138] [Example] [7] [:]5-(6-fluoro-4-((((4-methoxy-5-methylpyridin-2-yl)amino)methyl)-1H-indazol-7-yl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide [] []

[0139] The title compound, a white solid, was prepared in 23.80% overall yield using 4-methoxy-5-methylpyridine-2-amine from step 1, according to Example 1. MS: m / z: C 17H 17FN 6O 4S [M+H]+, calculated value 421; experimental value 421. 1H NMR (500 MHz, DMSO-d6) δ 13.28 (s, 1H), 8.99 (s, 1H), 8.23 ​​(s, 1H), 7.70 (s, 1H), 7.22–6.94 (m, 2H), 4.89–4.82 (m, 2H), 4.09 (s, 2H), 3.88 (s, 3H), 1.99 (s, 3H). []

[0140] Preparative HPLC purification conditions: XBridge C18 column, 19*200 mm, 5 μM; mobile phase A: ACN / H2O with 10 mM AA (5:95); mobile phase B: ACN / H2O with 10 mM AA (95:5); flow rate: 20 ml / min; gradient: 0% B to 40% B, 40% B over 20 min; wavelength: 220 nm. []

[0141] [Example] [8] [:]6-[[6-fluoro-7-(1,1,4-trisyloxy-1,2,5-thiadiazolin-2-yl)-1H-indazol-4-yl]methylamino]-4-methylpyridine-3-carboxynitrile [] []

[0142] The title compound, a white solid, was prepared in 17.62% overall yield using 6-amino-4-methyl-pyridine-3-carboxynitrile from step 1, according to Example 1. MS: m / z: C 17H 14FN 7O 3S [M+H]+ calculated value 416; experimental value 416. ¹H NMR (400 MHz, DMSO-d 6) δ 13.29 (s, 1H), 8.95–7.77 (m, 3H), 6.93 (d, J = 11.3 Hz, 1H), 6.56 (s, 1H), 4.86 (s, 2H), 4.41 (s, 2H), 2.28 (d, J = 4.4 Hz, 3H). []

[0143] Preparative HPLC purification conditions: Aeris PEPTIDE 5 μm XB-C18 Axia, 21.2 mm × 250 mm, 5 μM; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 50% B to 70% B, 70% B over 6.6 min; wavelength: 254 / 210 nm. []

[0144] [Example] [9] [:]5-[6-fluoro-4-[[(4-methoxy-6-methyl-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-dioxy-1,2,5-thiadiazolidine-3-one [] []

[0145] [process] [3] [:] []

[0146] [step] [1] [:] To 6-fluoro-2-[(4-methoxyphenyl)methyl]-7-(1,1,4-trisyloxy-1,2,5-thiadiazolin-2-yl)indazole-4-carboxaldehyde ( [Int-2] (50 mg, 0.12 mmol) and 4-methoxy-6-methylpyridin-2-amine (25 mg, 0.18 mmol) were added to a stirred mixture in a DCE (4 mL) with Ti(i-PrO)4 (68 mg, 0.24 mmol). The resulting mixture was stirred for 1 h at room temperature. NaBH3CN (31 mg, 0.48 mmol) was added to the mixture at 0 °C. The resulting mixture was stirred for 1 h at room temperature. After the reaction was complete as monitored by LCMS, the mixture was concentrated. The residue was purified by reverse-phase column chromatography to give a pale yellow solid, 5-[6-fluoro-4-[[(4-methoxy-6-methyl-2-pyridinyl)amino]methyl]-2-[(4-methoxyphenyl)methyl]indazol-7-yl]-1,1-dioxy-1,2,5-thiadiazolin-3-one (44 mg, 0.08 mmol, 68% yield). MS: m / z: C 25H 25FN 6O 5S [M+H]+, calculated value 541; experimental value 541.

[0147] [step] [2] [:] The title compound, a white solid, was prepared in step 2 in 10.00% yield according to the preparation method of Example 1. MS: m / z: C 17H 17FN 6O 4S [M+H]+ calculated value 421; experimental value 421. 1H NMR (400 MHz, DMSO- d 6) δ 13.24 (s, 1H), 12.68 (s, 1H), 8.29 (s, 2H), 7.08 (d, J = 11.5 Hz, 1H), 6.50 (s, 1H), 6.35 (s, 1H), 4.92 (d, J = 5.9 Hz, 2H), 4.12 (d, J = 4.9 Hz, 2H), 3.86 (s, 3H), 2.40 (s, 3H).

[0148] Preparative HPLC purification conditions: SunFire preparative C18 OBD column, 19*150 mm, 5 μm; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 40% B to 60% B, 60% B over 6.5 min; wavelength: 254 / 210 nm.

[0149] [Example]

[10] : 5-[6-fluoro-4-[[(3-methyl-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disidel-1,2,5-thiadiazolidin-3-one

[0150] The title compound, a white solid, was prepared in 6.21% overall yield using 3-methylpyridine-2-amine from step 1, according to Example 9. MS: m / z: C 16H 15FN 6O 3S [M+H]+, calculated value 391; experimental value 391. ¹H NMR (400 MHz, DMSO-d6) δ 13.26 (s, 1H), 8.59 (s, 1H), 8.26 (s, 1H), 8.84–8.65 (m, 2H), 6.92–6.71 (m, 2H), 4.85 (s, 2H), 4.06 (s, 2H), 2.23 (s, 3H).

[0151] Preparative HPLC purification conditions: SunFire preparative C18 OBD column, 19*150 mm, 5 μm; mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 60 mL / min; gradient: 5% B to 20% B, 20% B over 6.5 min; wavelength: 254 / 210 nm.

[0152] Example compounds prepared by the aforementioned procedure are listed in Table 1. [, , ] [] [surface] [1] Compound numbering structure Chemical name [Example] [1] 5-[6-fluoro-4-[[(6-methoxy-2-pyridinyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one [Example] [2] 5-(6-fluoro-4-(((5-methoxypyridin-2-yl)amino)methyl)-1H-indazol-7-yl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide [Example] [3] 5-[6-fluoro-4-[[(4-methoxy-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one [Example] [4] 5-[4-[[(4-cyclopropyl-2-pyridinyl)amino]methyl]-6-fluoro-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one [Example] [5] 5-[6-fluoro-4-[[(4-methyl-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one [Example] [6] 5-(4-(((4,6-dimethylpyridin-2-yl)amino)methyl)-6-fluoro-1H-indazol-7-yl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide [Example] [7] 5-(6-fluoro-4-(((4-methoxy-5-methylpyridin-2-yl)amino)methyl)-1H-indazol-7-yl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide [Example] [8] 6-[[6-fluoro-7-(1,1,4-trilateral-1,2,5-thiadiazolidin-2-yl)-1H-indazol-4-yl]methylamino]-4-methylpyridine-3-carboxynitrile [Example] [9] 5-[6-fluoro-4-[[(4-methoxy-6-methyl-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one [Example]

[10] 5-[6-fluoro-4-[[(3-methyl-2-pyridyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideloxy-1,2,5-thiadiazolidin-3-one

[0153] [Bioanalysis] [] The pharmacological properties of the compounds of the present invention can be confirmed by several bioanalyses known in this art. The bioanalyses exemplified below have been performed on the compounds of the present invention. Information relating to the preferred embodiments is shown in Table 2.

[0154] [PhosphoSens] [analyze] [] PhosphoSens® kinase assays were performed as described by the supplier (AssayQuant Technologies, Marlborough, MA). In short, 1000× compound solutions were prepared in DMSO in a 384-well reagent tray by serially diluting 10 mM DMSO stock solution at 3-fold intervals. Then, 50 nL of the compound dilution series was added to the corresponding wells of the 384-well assay tray. 40 mL of 1× assay buffer (50 mM HEPES pH 7.5, 500 µM EGTA, 10 nM MgCl2, 0.01% Brij-35, 1% glycerol, 1 mM DTT, and 0.2 mg / mL BSA) containing 1.25× acceptor (AQT0264) was transferred to each well of the assay tray to achieve a final acceptor concentration of 20 µM. Finally, 10 mL of 5× PTPN2 enzyme stock solution was added to each well of the assay tray, resulting in a final enzyme concentration of 150 pM. The reaction process curve was collected by sampling the fluorescence intensity every 71 seconds at room temperature with an excitation wavelength of 360 nm (λ ex360) and an emission wavelength of 480 nm (λ em480) for one hour.

[0155] [use] [DIFMUP] [Analysis of phosphatase activity as a substrate] [:] The following biochemical analysis of PTPN2 was performed: a 5× human PTPN2 stock solution (SRP5075, MilliporeSigma, Burlington, MA) and a 1.25× DiFMUP stock solution (D6567, ThermoFisher Scientific, Waltham, MA) were prepared in a 1× reaction buffer consisting of 50 mM HEPES, pH 7.4, 1 mM EDTA, 150 mM NaCl, 0.2 mg / mL BSA, 100 U / mL catalase, and 10 mM DTT. 40 mL of DiFMUP acceptor solution (final concentration 25 mM DiFMUP acceptor) was added to a Corning 3574 384-well white unbound surface microtiter containing 0.05 mL of serially diluted test compound prepared in DMSO. The reaction was initiated by adding 10 mL of enzyme solution, with a final PTPN2 concentration of 0.15 nM. Monitoring was performed every 105 seconds for 60 minutes at room temperature using a BioTek Synergy HTX disk reader (Agilent Technologies, Santa Clara, CA) at λ EX360 / λ EM460. The slope was obtained from the initial linear portion of the process curve fitted with a linear equation, and the 100% activity value of the untreated control was converted to inhibition %. The IC50 values ​​for each compound were obtained by fitting inhibition-compound concentration % curves using Dotmatics software (Dotmatics, Bishops Stortford, Hertfordshire, England).

[0156] [Cell proliferation analysis protocol] [] B16-F10 cells (ATCC, Manassas, VA, #CRL-6475) were cultured in DMEM growth medium (ThermoFisher Scientific, Waltham, MA, #11995-040) supplemented with 10% heat-inactivated FBS (ThermoFisher Scientific, #16140-071) and 1% penicillin / streptomycin (ThermoFisher Scientific, #15140-122). Cells were seeded at a density of 100 cells / well in 20 μL of total volume into two opaque 384-well microplates treated with tissue culture (PerkinElmer, Waltham, MA, #6007688) and incubated overnight at 37°C and 5% CO2. Then, 30 nL of the compound dissolved in DMSO was transferred from the source plate to the target wells using an Echo 650 acoustic liquid processor (Beckman Coulter, Indianapolis, IN). Negative control wells received only 30 nL of DMSO (0.15% final concentration). The culture dishes were returned to the incubator for 1 hour, and then cells were treated with either 5 μL of growth medium or 5 μL of growth medium containing 50 ng / mL recombinant mouse IFN-γ protein (R&D Systems, Minneapolis, MN, #485-MI / CF, 10 ng / mL final concentration) using an automated aspiration platform (INTEGRA Biosciences, Hudson, NH). The culture dishes were incubated at 37°C for 4 days, and cell proliferation was analyzed using CellTiter-Glo reagent (Promega, Madison, WI, #G7573, 25 μL / well). The intensity of the cold light signal was collected 15 minutes after CellTiter-Glo reagent addition using an EnVision 2105 disc reader (PerkinElmer) and analyzed using the Dotmatics software platform to calculate the IC50 value of the compound. We can identify off-target compound-mediated cytotoxicity by examining growth inhibition in the absence of IFNg.

[0157] [Phosphate group] [-STAT1] [Analysis Solution] [] B16-F10 cells (ATCC, Manassas, VA, #CRL-6475) were cultured in DMEM growth medium (ThermoFisher Scientific, Waltham, MA, #11995-040) supplemented with 10% heat-inactivated FBS (ThermoFisher Scientific, #16140-071) and 1% penicillin / streptomycin (ThermoFisher Scientific, #15140-122). Cells were seeded at a total volume of 20 μL at a density of 10,000 cells / well in opaque white 384-well microplates treated with tissue culture (PerkinElmer, Waltham, MA, #6007688) and incubated overnight at 37°C and 5% CO2. Then, 30 nL of the compound dissolved in DMSO was transferred from the source plate to the target wells using an Echo 650 acoustic liquid processor (Beckman Coulter, Indianapolis, IN). Negative control wells received only 30 nL of DMSO (0.15% final concentration). The culture trays were returned to the incubator for 1 hour, and then cells were treated with either 5 μL of growth medium or 5 μL of growth medium containing 500 ng / mL recombinant mouse IFN-γ protein (R&D Systems, Minneapolis, MN, #485-MI / CF, 100 ng / mL final concentration) using an automated aspiration platform (INTEGRA Biosciences, Hudson, NH). The culture trays were incubated at 37°C for 1 hour, and the phosphorylated STAT1 protein content was analyzed according to the manufacturer's instructions using a phosphate-STAT1 (Tyr701) HTRF kit (Cisbio, Bedford, MA, #63ADK026PEH). After 24 hours, HTRF signal intensity was collected using an EnVision 2105 tray reader (PerkinElmer) and analyzed using the Dotmatics software platform to calculate the compound's IC50 value.

[0158] [Bioanalytical Data] Table 2 provides an overview of the bioanalytical data for the examples / precedents prepared. For IC50 data, high DDT concentrations and / or DiFMUP assays were used; either assay may be used by those skilled in the art. Columns or rows marked with double asterisks indicate that an IC50 value or example has been provided. [, , ] [] [surface] [2] Instance number structure PTPN2 BChem HIGHDTT IC50 (uM) PTPN2 BChem DIFMUP IC50 (uM) pSTAT1 HTRF B16 EC50 (uM) Prolif. 5d B16 GI50 (uM) 1 0.005 0.002 0.287 0.160 2 0.042 0.024 1.880 0.880 3 0.006 0.009 0.764 0.350 4 0.001 0.002 3.115 2.689 5 0.005 0.008 0.440 0.166 6 ** 0.083 3.687 3.264 7 ** 0.199 >15 >15 8 ** 0.067 9.211 >15 9 ** 0.001 0.091 0.020 10 0.016 14.046 3.970

Claims

1. A compound having the following structure: or a pharmaceutically acceptable salt, stereoisomer, or hydrate thereof, wherein, Independently, each time it appears: R1 is selected from the group consisting of: −H, C1-12 alkyl substituted C1-12 alkyl, C1-3 alkoxy, amine, halogen, 5 to 10 aryl, −CN, cyclopropoxy, -OH and C3-6 cycloalkyl; R2 is selected from the group consisting of: −H, C1-12 alkyl, −CN, C3-6 cycloalkyl, 5 to 10 aryl, substituted C1-12 alkyl, C1-3 alkoxy, -OH and benzoxy; R3 is selected from the group consisting of: −H, C1-12 alkyl, substituted C1-12 alkyl, amine, halogen, −CN, −OCHF2, C1-3 alkoxy, -OH and α-lin-4-yl; The R4 group is selected from the group consisting of: −H, C1-12 alkyl, −OCH3, -OH and −CF3; and the R5 group is selected from the group consisting of: -H, C3-6 cycloalkyl, C1-12 alkyl and substituted C1-12 alkyl.

2. The compound of claim 1 or its pharmaceutically acceptable salt, stereoisomer, or hydrate, wherein: The R1 group is selected from the following groups: −CH2CH3, −CF3, −OCH3, secondary amines, tertiary amines, -Ph, cyclopropyl and cyclohexyl; the R2 group is selected from the following groups: -CH(CH3)2, -C(CH3)3, −CF3, −C(CH3)2R7 and −OCH3; the R3 group is selected from the following groups: −CH2CH3, −CHF2, secondary amines, tertiary amines and −OCH3; and the R4 group is −CH2CH3.

3. The compound of claim 1 or its pharmaceutically acceptable salt, stereoisomer, or hydrate, wherein: R1 and R3 are each independently selected from the following groups: -N(CH3)2 and -NHCH3.

4. The compound of claim 1 or its pharmaceutically acceptable salt, stereoisomer, or hydrate, wherein: R1 is selected from the group consisting of -H, -CH3, -OCH3 and cyclopropyl; R2 is selected from the group consisting of -H, -CH3, -CN and -OCH3; R3 is selected from the group consisting of -H, -CH3 and -OCH3; and R4 is selected from the group consisting of -H and -CH3.

5. The compound of claim 1 or its pharmaceutically acceptable salt, stereoisomer, or hydrate, wherein: R1 is cyclopropyl; R2 is −H; R3 is −H; R4 is −H; and R5 is -H.

6. The compound of claim 1 or its pharmaceutically acceptable salt, stereoisomer, or hydrate, wherein: R1 is −OCH3; R2 is C1-12 alkyl; R3 is −H; R4 is −H; and R5 is -H.

7. The compound of claim 1 or its pharmaceutically acceptable salt, stereoisomer, or hydrate, wherein: R1 is −OCH3; R2 is −H; R3 is −H; R4 is −H; and R5 is -H.

8. The compound of claim 1 or its pharmaceutically acceptable salt, stereoisomer, or hydrate, wherein: R1 is −OCH3; R2 is −H; R3 is C1-12 alkyl; R4 is −H; and R5 is -H.

9. The compound of claim 1 or its pharmaceutically acceptable salt, stereoisomer, or hydrate, wherein: R1 is −H; R2 is −OCH3; R3 is −H; R4 is −H; and R5 is -H.

10. The compound of claim 1 or its pharmaceutically acceptable salt, stereoisomer, or hydrate, wherein: R1 is a C1-12 alkyl group; R2 is −CN; R3 is −H; R4 is −H; and R5 is -H.

11. The compound of claim 1 or its pharmaceutically acceptable salt, stereoisomer, or hydrate, wherein: R1 is −H; R2 is −H; R3 is −H; R4 is a C1-12 alkyl group; and R5 is −H.

12. The compound of claim 1 or its pharmaceutically acceptable salt, stereoisomer, or hydrate, wherein: R1 is −H; R2 is −H; R3 is −OCH3; R4 is −H; and R5 is -H.

13. A compound or a pharmaceutically acceptable salt, stereoisomer, or hydrate thereof, wherein the compound is selected from the group consisting of: 5-[6-fluoro-4-[[(6-methoxy-2-pyridinyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideoxy-1,2,5-thiadiazolidine-3-one; 5-(6-fluoro-4-(((5-methoxypyridinyl)amino)methyl)-1H-indazol-7-yl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-[6-fluoro-4-[[(4-methoxy-2-pyridinyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideoxy-1,2,5-thiadiazolidine-3-one; 5-[4-[[(4-cyclopropyl-2-pyridinyl)amino]methyl]-6-fluoro-1H-indazol-7-yl]-1,1-disideoxy-1,2,5-thiadiazolidine-3-one; 5-[6-fluoro-4-[[(4-methyl-2-pyridinyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideoxy-1,2,5-thiadiazolidine-3-one; 5-(4-(((4,6-dimethylpyridin-2-yl)amino)methyl)-6-fluoro-1H-indazol-7-yl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(6-fluoro-4-(((4-methoxy-5-methylpyridin-2-yl)amino)methyl)-1H-indazol-7-yl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide; 6-[[6-fluoro-7-(1,1,4-trilateral-oxy-1,2,5-thiadiazolidin-2-yl)-1H-indazol-4-yl]methylamino]-4-methyl-pyridin-3-carboxynitrile; 5-[6-fluoro-4-[[(4-methoxy-6-methyl-2-pyridinyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideoxy-1,2,5-thiadiazolidin-3-one; and 5-[6-fluoro-4-[[(3-methyl-2-pyridinyl)amino]methyl]-1H-indazol-7-yl]-1,1-disideoxy-1,2,5-thiadiazolidin-3-one.

14. A pharmaceutical composition comprising a compound as claimed in any one of claims 1 to 13, or a pharmaceutically acceptable salt, stereoisomer, or hydrate thereof, and at least one pharmaceutically acceptable carrier.

15. Use of a compound of any one of claims 1 to 13, or a pharmaceutically acceptable salt, stereoisomer, or hydrate thereof, or a pharmaceutical composition of claim 14, for the preparation of a medicament for the treatment of cancer.

16. As claimed in claim 15, wherein the cancer is selected from: carcinomas, sarcomas, lymphomas, leukemia, melanoma, solid lymphoma, kidney cancer, breast cancer, lung cancer, bladder cancer, colon cancer, ovarian cancer, prostate cancer, pancreatic cancer, stomach cancer, brain cancer, head and neck cancer, skin cancer, uterine cancer, testicular cancer, glioma, esophageal cancer, liver cancer, and multiple myeloma.

17. As claimed in claim 16, wherein the cancer is selected from: adenocarcinomas, papillary adenocarcinomas, B-cell acute lymphoblastic lymphomas, non-Hodgkin's lymphomas, Burkitt's lymphomas, small lymphomas, Hodgkin's lymphomas, and hepatocarcinomas.

18. As used in claim 17, wherein the adenocarcinoma is papillary adenocarcinoma.

19. Use of a compound of any one of claims 1 to 13, or a pharmaceutically acceptable salt, stereoisomer, or hydrate thereof, or a pharmaceutical composition of claim 14, for the preparation of a medicament for the treatment of cancer, wherein the medicament is used in combination with an additional therapeutic agent.

20. As requested in claim 19, wherein the additional therapeutic agent is an immunotherapy agent.

21. As claimed in claim 20, wherein the immunotherapeutic agent is selected from the group consisting of: anti-PD-1 antibody, anti-PD-L1 antibody and anti-CTLA-4 antibody.

22. The use of any of claims 15 to 21, wherein the treatment includes radiation, surgery, chemotherapy or administration of biological drugs.

23. As claimed in claim 22, wherein the treatment comprises administering a biological agent that stimulates the immune system.

24. The use as claimed in claim 23, wherein the treatment comprises administration of an inhibitor of DGKα and / or DGKζ, an antagonist of the PD1 / PD-L1 axis, or an antagonist of CTLA4.

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