Protein tyrosine phosphatase inhibitors, compositions and methods of use

Abstract

Disclosed are compounds of Formula (I), pharmaceutically acceptable salts thereof, pharmaceutical compositions thereof, and combinations thereof, and methods of using the same as protein tyrosine phosphatase (PTPN2) inhibitors. These compounds are useful in the treatment of cancers and diseases susceptible to PTPN2 inhibition. # imgabs0 #

Description

[0001] Cross - reference to related applications

[0002] This application claims the priority of U.S. Provisional Application Serial No. 63 / 476,513, filed on December 21, 2022, the entire content of which is incorporated herein by reference. Technical field

[0003] This application discloses compounds, their pharmaceutically acceptable salts, their pharmaceutical compositions and combinations thereof, and methods of using them as inhibitors of protein tyrosine phosphatases. Background of the invention

[0005] Immune checkpoint blockade (ICB) is an innovative immunotherapy that targets immune - evasion mechanisms to improve the clinical response of cancer patients. For example, in the treatment of various types of cancer, checkpoint - blocking antibodies target cytotoxic T - lymphocyte antigen 4 (CTLA - 4), programmed cell death 1 (PD - 1) and its ligands, such as programmed cell death ligand 1 (PD - L1), thus significantly improving the treatment and survival outcomes of patients with these malignancies.

[0006] However, most patients receiving ICB treatment are either resistant to the treatment or eventually develop drug resistance. Specifically, mutations or deletions in the interferon - γ (IFNγ) signaling pathway are 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 activator of transcription (JAK / STAT) signaling. In addition, IFNγ indirectly restricts tumor growth by promoting the up - regulation of major histocompatibility complex class I (MHC - I), thereby enabling the presentation of antigens (Ag) to T cells. In vivo CRISPR screening using syngeneic mouse models has revealed the enrichment of the IFNγ pathway in anti - PD - 1 - resistant tumors. These studies have identified the above - mentioned IFNγ pathway members (JAK1 / 2 and STAT1) and interferon - γ receptors (IFNGR1 / IFNGR2) as resistance targets, and have also discovered some newly identified negative regulators, such as PTPN2 and Apelin receptor (APLNR), which represent new therapeutic targets (Charles Sinclair et al., Emerg Top LifeSci. (2021) 5(5):675 - 680).

[0007] Summary data from in vivo gene screening using CRISPR-Cas9 genome editing technology to identify genes contributing to checkpoint blockade resistance indicate that deletion of the protein tyrosine phosphatase (PTPN2) gene in tumor cells can enhance the efficacy of immunotherapy. The PTPN2 gene encodes a protein tyrosine phosphatase that regulates a range of intracellular processes. Deletion 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 PTPN2 therapeutic inhibition may enhance the effectiveness of immunotherapies that elicit an IFNγ response (Manguso, Robert T et al., Nature Vol. 547, 7664 (2017): 413-418).

[0008] Non-receptor protein tyrosine phosphatase type 2 (PTPN2), also known as T cell protein tyrosine phosphatase (TCPTP), is an intracellular member of the class I subfamily of phosphotyrosine-specific phosphatases that controls multiple cellular regulatory processes by removing phosphate groups from tyrosine substrates. PTPN2 is widely expressed, but is most highly expressed in hematopoietic and placental cells (Mosinger, B. Jr et al., Proc Natl Acad Sci USA (1992) 89:499-503). In humans, the expression of PTPN2 is controlled post-transcriptionally by two splice variants: one is a 45 kDa splice variant that contains a nuclear localization signal at the C-terminus upstream of the splice junction; the other is a typical 48 kDa splice variant that contains an endoplasmic reticulum retention motif at the C-terminus (Tillmann U. et al., Mol Cell Biol (1994) 14:3030-3040). Under certain cellular stress conditions, the 45 kDa isoform can be passively transported to the cytosol. Both isoforms possess an N-terminal phosphotyrosine phosphatase catalytic domain and, as a key negative regulator of the JAK-STAT pathway, PTPN2 directly regulates signaling through cytokine receptors. The PTPN2 catalytic domain has 74% sequence homology with PTPN1 (also known as PTP1B) and has similar enzyme kinetics (Romsicki Y. et al., Arch Biochem Biophys (2003) 414:40-50).

[0009] The T cell protein tyrosine phosphatase PTPN2 has been further confirmed 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 inactivates Src family kinases, thereby regulating T cell responses. PTPN2 deficiency has been shown to lower the threshold of TCR-dependent CD8 + T cell proliferation in vivo. Consistent with these findings, T cell-specific PTPN2-deficient mice have been shown to develop widespread inflammation and autoimmunity. This autoimmunity is associated with elevated levels of pro-inflammatory cytokines in the serum, increased anti-nuclear antibodies, T cell infiltration in non-lymphoid tissues, and liver disease. These data further suggest that PTPN2 is a key negative regulator of TCR signaling that sets the threshold of TCR-induced naive T cell responses to prevent autoimmune and inflammatory diseases.

[0010] In addition to PTPN2 encoding the T cell PTP (TCPTP) as a susceptibility gene locus for autoimmune diseases, SNPs in PTPN2 are also 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).

[0011] The above research results indicate that inhibiting PTPN2 is a potential therapeutic strategy to improve the efficacy of cancer treatment regimens related to ICB resistance. Summary of the Invention

[0013] The present invention relates to compounds, pharmaceutically acceptable salts thereof, pharmaceutical compositions thereof, and combinations thereof, which are potent inhibitors of protein tyrosine phosphatases, such as non-receptor protein tyrosine phosphatase type 2 (PTPN2) and / or non-receptor protein tyrosine phosphatase type 1 ((PTPN1), also known as protein tyrosine phosphatase-1B (PTP1B)). The present invention further provides methods for treating, preventing, or ameliorating cancer, including administering to an individual in need an effective amount of the PTPN2 / PTPN1 inhibitor disclosed herein. In a preferred embodiment, the compounds have a monocyclic core structure compared to the compounds reported in the literature, wherein the compounds contain a fused bicyclic core.

[0014] In some embodiments, inhibitors of a protein tyrosine phosphatase (e.g., PTPN2 and / or PTP1B) are disclosed herein, which comprise a compound disclosed herein, such as a compound of formula (I). In other embodiments, methods of treating a disease or disorder are disclosed herein, such as cancer, type 2 diabetes, obesity, metabolic diseases, or any other disease, disorder, or affliction that responds favorably to treatment with a PTPN2 or PTP1B inhibitor, including 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 expanded form in the present invention.

[0015] The first aspect of the invention provides at least one compound of formula (I) having the following structure:

[0016]

[0017] wherein, each occurrence independently:

[0018] R 1 is selected from 1H-1,2,4-triazol-3-yl,

[0019]

[0020] R 2 is selected from -H, alkyl, and substituted alkyl;

[0021] R 3 is selected from -H, alkyl, and substituted alkyl;

[0022] R 4 is selected from -H, alkyl, substituted alkyl, branched alkyl, alkoxy, halogen, cyano, amine, hydroxy, aryl, and substituted aryl;

[0023] R 5 is selected from -H, alkyl, substituted alkyl, branched alkyl, alkoxy, halogen, cyano, amine, hydroxy, aryl, and substituted aryl.

[0024] Compounds selected from the following are further disclosed:

[0025] 5-(4-(((4-(2,4-dimethylphenyl)thiazol-2-yl)(methyl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0026] 5-(4-(((1H-pyrazol-3-yl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0027] 5-(2-Fluoro-6-hydroxy-4-((isoxazol-3-ylamino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0028] 5-(2-Fluoro-6-hydroxy-4-((((5-methylisoxazol-3-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0029] 5-(4-((((1H-1,2,4-triazol-3-yl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0030] 5-(2-Fluoro-6-hydroxy-4-((((1-methyl-1H-pyrazol-3-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0031] 5-(4-((((1,3,4-thiadiazol-2-yl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0032] 5-(2-Fluoro-6-hydroxy-4-((((5-methyl-1H-imidazol-2-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0033] 5-(2-Fluoro-6-hydroxy-4-((thiazol-2-ylamino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0034] 5-(4-((((1H-imidazol-2-yl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0035] 5-(2-Fluoro-6-hydroxy-4-((((1-methyl-1H-imidazol-2-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0036] 5-(2-Fluoro-6-hydroxy-4-((((5-methyl-1,3,4-thiadiazol-2-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0037] 5-(2-Fluoro-6-hydroxy-4-((((4-methylthiazol-2-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0038] 5-(2-Fluoro-6-hydroxy-4-(((3-methylisothiazol-5-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0039] 5-(2-Fluoro-6-hydroxy-4-(((3-methyl-1H-pyrazol-5-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0040] or a pharmaceutically acceptable salt thereof.

[0041] In some embodiments, a compound of formula (I) is formulated into a pharmaceutically acceptable composition comprising the compound of formula (I) and a pharmaceutically acceptable carrier.

[0042] Also disclosed herein is a method of treating cancer in a patient in need thereof, comprising administering to the patient a combination of 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. By way of example, in some embodiments, the immunotherapeutic agent is an antibody.

[0043] Also disclosed herein is a method of treating cancer in a patient in need thereof, comprising administering to the patient an effective amount of a compound disclosed herein, such as a compound of formula (I).

[0044] Further disclosed herein is a method of treating a metabolic disease in a patient in need thereof, comprising administering to the patient an effective amount of a compound disclosed herein, such as a compound of formula (I).

[0045] In some embodiments, the method comprises treating cancer. In some embodiments, the cancer comprises pancreatic cancer, breast cancer, multiple myeloma, melanoma or secretory cell carcinoma.

[0046] Also disclosed herein is a composition for treating cancer in a patient in need thereof, wherein the composition comprises a combination of a compound disclosed herein (such as a compound of formula (I)) and an additional therapeutic agent. In some embodiments, the additional therapeutic agent is an immunotherapeutic agent. By way of example, in some embodiments, the immunotherapeutic agent is selected from anti-PD-1 antibodies and anti-PD-L1 antibodies.

[0047] Further disclosed herein is a composition for treating a metabolic disease in a patient in need thereof, wherein the composition comprises a compound disclosed herein, such as a compound of formula (I). DETAILED DESCRIPTION OF THE INVENTION

[0049] The present invention relates to compounds, pharmaceutically acceptable salts thereof, pharmaceutical compositions thereof and combinations thereof, said compounds being potent inhibitors of protein tyrosine phosphatases, such as non-receptor protein tyrosine phosphatase type 2 (PTPN2) and / or non-receptor protein tyrosine phosphatase type 1 (PTPN1), also known as protein tyrosine phosphatase-1B (PTP1B). The present invention further provides methods of treating, preventing or ameliorating cancer, comprising administering to an individual in need thereof an effective amount of a PTPN2 / PTPN1 inhibitor disclosed herein. In a preferred embodiment, the compounds have a monocyclic core structure as compared to compounds reported in the literature, wherein the compounds contain a fused bicyclic core.

[0050] Definitions

[0051] Chemical Definitions

[0052] The definitions of specific functional groups and chemical terms are described in more detail below. 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. In addition, general principles of organic chemistry and specific functional moieties and reactivity are described in: 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.

[0053] The abbreviations used herein have their conventional meanings in the chemical and biological arts. The chemical structures and chemical formulas listed herein are constructed in accordance with the standard valence rules known in the chemical art.

[0054] The compounds described herein may contain one or more asymmetric centers and thus may exist in various isomeric forms, such as enantiomers and / or diastereomers. For example, the compounds described herein may be in the form of individual enantiomers, diastereomers, geometric isomers, or mixtures of stereoisomers, including racemic mixtures and mixtures enriched in one or more stereoisomers. The isomers can be separated from the mixture by methods known to those skilled in the art, including chiral high performance liquid chromatography (HPLC) and the formation and crystallization of chiral salts; or the preferred isomers can 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 at page 268 (E.L. Eliel, ed., Univ. of Notre Dame Press, Notre Dame, IN 1972). The present disclosure also encompasses the compounds described herein in the form of individual isomers or mixtures of multiple isomers that are substantially free of other isomers.

[0055] In the compositions provided herein, enantiomerically pure compounds may coexist with other active or inactive ingredients. For example, a pharmaceutical composition containing an enantiomerically pure R-compound may contain, for example, approximately 90% excipient and approximately 10% enantiomerically pure R-compound.

[0056] Those skilled in the art can more readily understand the features and advantages of the invention described in the present disclosure from the following definitions. Certain features of the invention described in the context of separate embodiments may also be combined to form a single embodiment, or extrapolated to include multiple embodiments. The embodiments identified herein as exemplary or preferred are illustrative and not restrictive.

[0057] Unless otherwise expressly stated herein, the singular forms also include the plural forms. For example, "a" can mean one or more than one.

[0058] The phrase "compound" as used herein refers to at least one compound. For example, a compound of formula (I) includes one compound of formula (I) and two or more compounds of formula (I).

[0059] Unless otherwise specified, any heteroatom with unsatisfied valency is assumed to have hydrogen atoms sufficient to satisfy the valency.

[0060] The definitions set forth herein prevail over those described in any patent, patent application, and / or patent application publication incorporated herein by reference.

[0061] The following are definitions of various terms used to describe the present invention. These definitions apply to the terms used throughout the specification (unless otherwise limited in specific instances), whether used alone or as part of a larger group.

[0062] Throughout the specification, those skilled in the art can select groups and their substituents to provide stable moieties and compounds.

[0063] In accordance with the convention in the art, in the structural formulas herein, is used to describe the bond that serves as the point of attachment of a moiety or substituent to the core or backbone structure.

[0064] As used herein, the terms “halo” and “halogenated” refer to F, Cl, Br, and I.

[0065] The term “cyano” refers to the group -CN.

[0066] The term “amino” refers to the group -NH2.

[0067] The term “oxo” refers to the group ═O.

[0068] As used herein, the term “alkyl” refers to branched and straight-chain saturated aliphatic hydrocarbon groups 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, sec-butyl, and tert-butyl), and pentyl (e.g., n-pentyl, isopentyl, 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” represents straight-chain and branched-chain alkyl groups having one to six carbon atoms.

[0069] 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, “C 1-4 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.

[0070] 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 C 1-4 cyanoalkyl.

[0071] 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 C 1-4 aminoalkyl.

[0072] The term "hydroxyalkyl" includes branched and straight-chain saturated alkyl groups substituted with one or more hydroxy groups. For example, "hydroxyalkyl" includes -CH2OH, -CH2CH2OH, and C 1-4 hydroxyalkyl.

[0073] The term "hydroxyfluoroalkyl" includes branched and straight-chain saturated alkyl groups substituted with one or more hydroxy groups and one or more fluorine atoms. For example, "hydroxyfluoroalkyl" includes -CHFCH2OH, -CH2CHFC(CH3)2OH, and C 1-4 hydroxyfluoroalkyl.

[0074] 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 saturated ring carbon atom. Representative examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclopentyl, and cyclohexyl. When a number appears in subscript form after the symbol "C", the subscript more specifically defines the number of carbon atoms that a particular cycloalkyl may contain. For example, "C3-C6 cycloalkyl" means a cycloalkyl having three to six carbon atoms.

[0075] As used herein, the term "heterocycle" refers to an organic compound having a cyclic structure with both carbon atoms and non-carbon atoms (such as oxygen, nitrogen).

[0076] As used herein, the term "alkoxy" refers to an alkyl group attached to the parent molecular moiety via an oxygen atom, such as methoxy (-OCH3). For example, "C 1-3 alkoxy" means an alkoxy having one to three carbon atoms.

[0077] As used herein, the term "alkoxyalkyl" refers to an alkoxy group attached to an alkyl group via its oxygen atom, and the alkyl group is attached to the parent molecular moiety, such as methoxymethyl (-CH2OCH3). For example, "C 2-4 alkoxyalkyl" means an alkoxyalkyl having two to four carbon atoms, such as -CH2OCH3, -CH2CH2OCH3, -CH2OCH2CH3, and -CH2CH2OCH2CH3.

[0078] As used herein, the term "amine" refers to a compound in which a nitrogen atom is directly bonded to a number of carbon atoms. Specific examples include derivatives of ammonia (-NH3) produced by the progressive replacement of three hydrogen atoms by hydrocarbon groups. Depending on the number of carbons bonded to the nitrogen atom, amines are classified as primary (1°), secondary (2°), or tertiary (3°). For example, a primary amine has one carbon bonded to the nitrogen (R-NH2), a secondary amine has two carbons bonded to the nitrogen (R2-NH), and a tertiary amine has three carbons bonded to the nitrogen (R3-N), where R is an alkyl group.

[0079] As used herein, the term "heteroaryl" refers to an aromatic heterocycle having 5 to 10 members and having at least one heteroatom selected from nitrogen, oxygen, and sulfur and containing at least 1 carbon atom, including monocyclic and bicyclic systems.

[0080] The phrase "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions, and / or dosage forms that, within the scope of reasonable medical judgment, are suitable for contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response, or other problems or complications and that have a reasonable benefit / risk ratio.

[0081] The compounds of formula (I) can be provided in the form of an amorphous solid or a crystalline solid. The compounds of formula (I) can be provided in the form of an amorphous solid by lyophilization.

[0082] It should also be understood that solvates (e.g., hydrates) of the compounds of formula (I) are also encompassed within the scope of the present invention. The term "solvate" refers to the physical association of a compound of formula (I) with one or more solvent molecules, whether organic or inorganic. Such physical association includes hydrogen bonding. In some cases, solvates can be isolated, for example when one or more solvent molecules are incorporated into the lattice of a crystalline solid. "Solvate" encompasses both the solution phase and isolable solvates. Exemplary solvates include hydrates, ethanolates, methanolates, isopropanolates, acetonitrile solvates, and ethyl acetate solvates. Solvation methods are known in the art.

[0083] The various forms of prodrugs are well known in the art and are described in the following:

[0084] a) The Practice of Medicinal Chemistry, Camille G. Wermuth et al., Ch 31, (Academic Press, 1996);

[0085] b) Design of Pro-drugs, H. Bundgaard, ed., (Elsevier, 1985);

[0086] 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

[0087] d) Hydrolysis in Drug and Prodrug Metabolism, Bernard Testa and Joachim M. Mayer, (Wiley-VCH, 2003).

[0088] In addition, after preparation, the compounds of formula (I) can be separated and purified to obtain a composition (“substantially pure”) in which the content of the compound of formula (I) is equal to or greater than 99% by weight, which can then be used or formulated as described herein. Such “substantially pure” compounds of formula (I) are also covered herein and are part of the present invention.

[0089] “Stable compound” and “stable structure” mean that the compound is stable enough to be separated to an appropriate purity from the reaction mixture and formulated into an effective therapeutic agent. The present invention is intended to embody stable compounds.

[0090] Those of ordinary skill in the art should also understand that the compounds described and claimed herein as embodiments of the present invention also exist in “tautomeric” forms. Herein, tautomers that exist in tautomeric forms refer to structural isomers that can be easily interconverted under rapid equilibrium. Herein, the process of interconversion is called “tautomerization”.

[0091] For example, in the following embodiments, the pyridone tautomers can be represented as follows:

[0092]

[0093] The disclosed structures can be easily interconverted between the structures shown on the left hand side and the structures shown on the right hand side.

[0094] “Therapeutically effective amount” is intended to include the amount of the compounds of the present invention used alone, or the amount of a combination of the claimed compounds, or the amount of a combination of the compounds of the present invention with other active ingredients that are effective as inhibitors or effective in treating or ameliorating cancer.

[0095] As used herein, "treatment" covers treating a disease state in a mammal, particularly a human, and includes: (a) preventing the mammal from developing the disease state, particularly when the mammal is predisposed to the disease state but has not been diagnosed as having the disease state; (b) inhibiting the disease state, i.e., arresting its development; and / or (c) alleviating the disease state, i.e., causing the disease state to regress.

[0096] The compounds of the invention are intended to include all isotopes of the atoms that occur in the compounds of the invention. Isotopes include those atoms having the same atomic number but different mass numbers. As general non-limiting examples, isotopes of hydrogen include deuterium (D) and tritium (T). Isotopes of carbon include 13 C and 14 C. The isotopically labeled compounds of the invention can generally be prepared by conventional techniques known to those skilled in the art or by methods similar to those described herein, using appropriately isotopically labeled reagents in place of the unlabeled reagents originally employed. For example, methyl (-CH3) also includes deuterated methyl, such as -CD3.

[0097] The term "pharmaceutically acceptable salts" is intended to include salts of the active compounds which are prepared with relatively non-toxic acids or bases, depending on the particular substituents of the compounds described herein. When the compounds of the present disclosure contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of the desired base, either neat or in a suitable inert solvent. Examples of pharmaceutically acceptable base addition salts include sodium, potassium, calcium, ammonium, organic amino, magnesium salts or the like.

[0098] As defined herein, terms such as "inhibit" with respect to protein-inhibitor (e.g., antagonist) interactions refer to a negative effect (e.g., reduction) on the activity or function of a protein relative to the protein activity or function in the absence of the inhibitor. In some embodiments, inhibition refers to a reduction in a disease or disease symptom. In some embodiments, inhibition refers to a reduction in the activity of a signal transduction pathway or signaling pathway. Thus, inhibition includes at least partially, partially or completely blocking a stimulus, reducing, preventing or delaying activation, or inactivating, desensitizing or downregulating signal transduction or enzyme activity or the amount of a protein. In some embodiments, inhibition refers to a reduction in the activity of a protein tyrosine phosphatase (e.g., non-receptor protein tyrosine phosphatase type 2 (PTPN2) or non-receptor protein tyrosine phosphatase type 1 (PTP1B)). Thus, inhibition can include at least partially, partially or completely reducing a stimulus; reducing or decreasing activation or inactivation; desensitizing or downregulating signal transduction or enzyme activity or the amount of a protein tyrosine phosphatase, such as non-receptor protein tyrosine phosphatase type 2 (PTPN2) or non-receptor protein tyrosine phosphatase type 1 (PTP1B).

[0099] A "patient" or "individual" in need is a living organism that has or is susceptible to a disease or condition that can be treated by administering a compound or pharmaceutical composition as provided herein. Non-limiting examples include humans, other mammals, bovines, rats, mice, dogs, monkeys, goats, sheep, 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 test 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 elderly animal. In some embodiments, the patient is an elderly human. In some embodiments, the patient is an elderly mammal. In some embodiments, the patient is an aged patient.

[0100] "Disease", "disorder", or "condition" refers to a state of life or health of a patient or individual that can be treated with a compound, pharmaceutical composition, or method provided herein. In some embodiments, the compounds and methods described herein include, for example, reducing or eliminating one or more symptoms of a disease, disorder, or condition 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.

[0101] As used herein, the term "signaling pathway" refers to a series of interactions between a cell and optionally present extracellular components (e.g., proteins, nucleic acids, small molecules, ions, lipids) that convey a change in one component to one or more other components, which in turn may convey the change to additional components, and the change is optionally propagated to other signaling pathway components.

[0102] "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 to the patient. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, standard saline solutions, lactated Ringer's solutions, standard sucrose, standard glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavoring agents, salt solutions (such as Ringer's solutions), alcohols, oils, gelatin, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxypropyl methylcellulose, polyvinylpyrrolidone, and pigments. Such formulations can be sterilized and, if desired, mixed with auxiliaries (such as lubricants, preservatives, stabilizers, wetting agents, emulsifying agents, salts for influencing osmotic pressure, buffers, coloring agents, and / or flavoring agents, etc.) that do not react unfavorably with the compounds of the present invention. Those skilled in the art will understand that other pharmaceutical excipients can also be used in the present disclosure.

[0103] The term "formulation" is intended to include capsules in which an active compound is formulated with an encapsulating material as a carrier, where the active ingredient (whether or not it includes other carriers) is encapsulated by the carrier and thus bound thereto.

[0104] Similarly, cachets and lozenges are also included. Tablets, powders, capsules, pills, cachets, and lozenges can be used as solid dosage forms suitable for oral administration.

[0105] As used herein, the term "administer" refers to oral administration to an individual, administration as a suppository, topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intracranial, intranasal, or subcutaneous administration, or implantation of a sustained release device, such as a micro-osmotic pump. Administration can be by any route, including parenteral and transmucosal (e.g., buccal, sublingual, transpalatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, for example, intravenous, intramuscular, intraarterial, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other delivery modes include, but are not limited to, the use of liposomal formulations, intravenous infusions, transdermal patches, etc. "Co-administration" refers to the administration of the compounds or compositions described herein simultaneously with, before, or after one or more other therapies (e.g., anti-cancer agents, chemotherapeutic agents, or immunotherapeutic agents). The compounds or compositions described herein can be administered alone or in combination to a patient. Co-administration includes the administration of the compounds or compositions alone or in combination (multiple compounds or agents) simultaneously or sequentially. Thus, if desired, the formulations can also be used in combination with other active substances (e.g., to reduce metabolic degradation).

[0106] The pharmaceutical compositions described herein can be prepared by any method known in the art of pharmacology. Generally, such methods of preparation include the steps of: mixing the disclosed compound (“active ingredient”) with a carrier and / or one or more other accessory ingredients, and then, if necessary and / or desired, shaping and / or encapsulating the product into the desired single-dose or multi-dose unit. The pharmaceutical compositions can be prepared, encapsulated, and / or sold in bulk, as a single unit dose, and / or as multiple single unit doses. As used herein, “unit dose” refers to an individual dose of a pharmaceutical composition that contains a predetermined amount of the active ingredient. The amount of the active ingredient is usually equal to the dose of the active ingredient to be administered to an individual and / or a suitable fraction of that dose, such as half or one-third of that dose.

[0107] Method of treatment

[0108] The present disclosure features compounds, compositions, and methods that comprise the compounds disclosed herein, such as compounds of formula (I). In some embodiments, the compounds, compositions, and methods disclosed herein are used for the prevention or treatment of a disease, disorder, or condition. Exemplary diseases, disorders, or conditions include, but are not limited to, cancer, type 2 diabetes, metabolic syndrome, obesity, or metabolic diseases.

[0109] Cancer

[0110] In some embodiments, the compounds disclosed herein (e.g., compounds of formula (I)) are used for the treatment of cancer. As used herein, "cancer" refers to human cancers and carcinomas, sarcomas, adenocarcinomas (e.g., papillary adenocarcinomas), lymphomas, leukemias, melanomas, etc., including solid cancers and lymph cancers, kidney cancer, breast cancer, lung cancer, bladder cancer, colon cancer, ovarian cancer, prostate cancer, pancreatic cancer, gastric cancer, brain cancer, head and neck cancer, skin cancer, uterine cancer, testicular cancer, glioma, esophageal cancer, liver cancer, including hepatocarcinoma, lymphoma, including B acute lymphoblastic lymphoma, non-Hodgkin's lymphomas (e.g., Burkitt's lymphomas, small cell lymphomas and large cell lymphomas), Hodgkin's lymphoma, leukemia (including AML, ALL and CML) and / or multiple myeloma. In some other cases, "cancer" refers to lung cancer, breast cancer, ovarian cancer, epithelial ovarian cancer, leukemia, lymphoma, melanoma, pancreatic cancer, sarcoma, bladder cancer, bone cancer, biliary tract cancer, adrenal cancer, salivary gland cancer, bronchial cancer, oral cancer, mouth cancer or pharyngeal cancer, laryngeal cancer, kidney cancer, gynecological cancer, brain cancer, central nervous system cancer, peripheral nervous system cancer, blood tissue cancer, small intestine cancer or appendiceal cancer, cervical cancer, colon cancer, esophageal cancer, gastric cancer, liver cancer, head and neck cancer, kidney cancer, myeloma, thyroid cancer, prostate cancer, metastatic cancer or carcinoma.

[0111] Exemplary cancers treatable with the compounds, pharmaceutical compositions or methods provided herein include lymphoma, B-cell lymphoma, heavy chain disease, alpha chain disease, gamma chain disease, mu chain disease, Waldenstrom's macroglobulinemia, benign monoclonal gammopathy, sarcoma, bladder cancer, bone cancer, brain cancer, cervical cancer, colon cancer, esophageal cancer, gastric cancer, head and neck cancer, kidney cancer, myeloma, thyroid cancer, leukemia, prostate cancer, 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-resistant breast cancer, ductal carcinoma of the breast, 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, pinealoma, ependymoma, oligodendroglioma, meningioma, glioma or melanoma. Further examples include thyroid cancer, cancers of the endocrine system, 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, gastric cancer, uterine cancer or neuroblastoma, Hodgkin's disease, non-Hodgkin's lymphoma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, immunocyte amyloidosis, ovarian cancer, rhabdomyosarcoma, essential thrombocythemia, primary macroglobulinemia, primary brain tumor, cancer, malignant insulinoma, malignant carcinoid, bladder cancer, precancerous skin lesions, testicular cancer, lymphoma, thyroid cancer, neuroblastoma, esophageal cancer, urogenital cancer, malignant hypercalcemia, endometrial cancer, adrenocortical cancer, endocrine or exocrine pancreatic neoplasms, medullary thyroid cancer, medullary carcinoma of the thyroid, melanoma, colorectal cancer, papillary thyroid cancer and hepatocellular carcinoma.

[0112] A first aspect of the invention provides at least one compound of formula (I) having the following structure:

[0113]

[0114] wherein, each occurrence independently:

[0115] R 1 is selected from 1H-1,2,4-triazol-3-yl,

[0116]

[0117] R2 Selected from -H, alkyl, and substituted alkyl;

[0118] R 3 Selected from -H, alkyl, and substituted alkyl;

[0119] R 4 Selected from -H, alkyl, substituted alkyl, branched alkyl, alkoxy, halogen, cyano, amine, hydroxy, aryl, and substituted aryl;

[0120] R 5 Selected from -H, alkyl, substituted alkyl, branched alkyl, alkoxy, halogen, cyano, amine, hydroxy, aryl, and substituted aryl.

[0121] In one embodiment of the compound of formula (I):

[0122] R 1 Selected from

[0123]

[0124] R 3 Selected from -H and -CH3;

[0125] R 4 Is -H and -CH3.

[0126] In another embodiment of the compound of formula (I):

[0127] R 1 Is

[0128] R 4 Selected from -H and -CH3.

[0129] In one embodiment of the compound of formula (I):

[0130] R 1 Is

[0131] R 4 Selected from -H, -CH3, and 2,4-dimethylphenyl;

[0132] R 5 Selected from -H and -CH3.

[0133] In another embodiment of the compound of formula (I):

[0134] R 1 Selected from

[0135]

[0136] R 4 Selected from -H and -CH3.

[0137] In one embodiment of the compound of formula (I):

[0138] R 1 is

[0139] R 3 selected from -H and -CH3;

[0140] R 4 is -H and -CH3;

[0141] R 5 is -H and -CH3.

[0142] In another embodiment, the compound is selected from

[0143] 5-(4-(((4-(2,4-dimethylphenyl)thiazol-2-yl)(methyl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0144] 5-(4-(((1H-pyrazol-3-yl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0145] 5-(2-fluoro-6-hydroxy-4-((isoxazol-3-ylamino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0146] 5-(2-fluoro-6-hydroxy-4-(((5-methylisoxazol-3-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0147] 5-(4-(((1H-1,2,4-triazol-3-yl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0148] 5-(2-fluoro-6-hydroxy-4-(((1-methyl-1H-pyrazol-3-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0149] 5-(4-(((1,3,4-thiadiazol-2-yl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide;

[0150] 5-(2-Fluoro-6-hydroxy-4-(((5-methyl-1H-imidazol-2-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide;

[0151] 5-(2-Fluoro-6-hydroxy-4-((thiazol-2-ylamino)methyl)phenyl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide;

[0152] 5-(4-(((1H-Imidazol-2-yl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide;

[0153] 5-(2-Fluoro-6-hydroxy-4-(((1-methyl-1H-imidazol-2-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide;

[0154] 5-(2-Fluoro-6-hydroxy-4-(((5-methyl-1,3,4-thiadiazol-2-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide;

[0155] 5-(2-Fluoro-6-hydroxy-4-(((4-methylthiazol-2-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide;

[0156] 5-(2-Fluoro-6-hydroxy-4-(((3-methylisothiazol-5-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide;

[0157] 5-(2-Fluoro-6-hydroxy-4-(((3-methyl-1H-pyrazol-5-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide;

[0158] or a pharmaceutically acceptable salt thereof.

[0159] In one embodiment, the present invention includes a pharmaceutical composition comprising a compound of formula (I), a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.

[0160] In another 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) 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, renal cancer, breast cancer, lung cancer, bladder cancer, colon cancer, ovarian cancer, prostate cancer, pancreatic cancer, gastric cancer, brain cancer, head and neck cancer, skin cancer, uterine cancer, testicular cancer, glioma, esophageal cancer, liver cancer, including hepatocellular carcinoma, lymphoma, including B acute lymphoblastic lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, small lymphoma, Hodgkin's lymphoma, leukemia, and multiple myeloma.

[0161] In another embodiment, the present invention includes a method for treating cancer in a patient in need thereof, comprising administering to the patient a combination of an effective amount of a compound of formula I and an additional therapeutic agent.

[0162] In one embodiment, the additional therapeutic agent is an immunotherapeutic agent.

[0163] In another embodiment, the immunotherapeutic agent is selected from anti-PD-1 antibodies, anti-PD-L1 antibodies, and anti-CTLA-4 antibodies.

[0164] In one embodiment, the method for treating cancer in a patient in need thereof comprises administering to the patient a pharmaceutically acceptable composition of an effective amount of a compound of formula I.

[0165] In another embodiment, the method for treating cancer is selected from radiation, surgery, chemotherapy, or administration of a biologic.

[0166] In one embodiment, the method for treating cancer is administration of a biologic, and the biologic is a drug that stimulates the immune system.

[0167] In another embodiment, the method for treating cancer comprises administering to an individual an inhibitor of DGKα and / or DGKζ, an antagonist of the PD1 / PD-L1 axis, and an antagonist of CTLA4.

[0168] These embodiments are not intended to limit the scope of the present invention.

[0169] Synthetic methods

[0170] The compounds of the present invention can be prepared by the methods and examples presented below and by methods known to those of ordinary skill in the art. In the following individual examples, unless indicated otherwise, the R groups are defined as above for each formula. The optimal reaction conditions and reaction times can vary depending on the reactants used. Unless otherwise stated, those of ordinary skill in the art can readily select solvents, temperature, pressure, and other reaction conditions.

[0171] The intermediates used in the following syntheses are commercially available or can be readily prepared by methods known to those skilled in the art. The progress of the reaction can be monitored by conventional methods such as thin layer chromatography (TLC) or high performance liquid chromatography - mass spectrometry (HPLC - MS). The intermediates and products can be purified by methods known in the art, including column chromatography, HPLC, preparative TLC or preparative HPLC.

[0172] Preparation of the key synthetic intermediate (Int - 2)

[0173] The preparation of 5-(4 - bromo - 2 - fluoro - 6 - ((4 - methoxybenzyl)oxy)phenyl)-1,2,5 - thiadiazolidine - 3 - one 1,1 - dioxide (Int2) is shown in Scheme 1.

[0174] Scheme 1:

[0175]

[0176] Step 1: Synthesis of 5 - bromo - 1 - fluoro - 3 - ((4 - methoxybenzyl)oxy)-2 - nitrobenzene (1 - 2)

[0177] Potassium carbonate (17.4 g, 126.06 mmol) was added portionwise to a stirred solution of 5 - bromo - 1,3 - difluoro - 2 - nitro - benzene (10 g, 42.02 mmol) and (4 - methoxyphenyl)methanol (6.1 g, 44.12 mmol) in DMF (100 mL) at room temperature. The resulting mixture was stirred overnight at 70 °C under a nitrogen atmosphere. TLC indicated completion of the reaction. The reaction mixture was diluted with water (300 mL) and extracted with ethyl acetate (3 × 300 mL). The combined organic layers were washed with brine and dried over anhydrous sodium sulfate. After filtration, the filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (ethyl acetate / petroleum ether = 1 / 20) to give the desired product 5 - bromo - 1 - fluoro - 3 - [(4 - methoxyphenyl)methoxy]-2 - nitro - benzene as a pale yellow solid (10 g, 66.8% yield).

[0178] Step 2: Synthesis of 4 - bromo - 2 - fluoro - 6 - ((4 - methoxybenzyl)oxy)aniline (1 - 3)

[0179] At room temperature, NH4Cl (15.16 g, 280.79 mmol) and Fe (15.68 g, 280.79 mmol) were added to a stirred solution of 5-bromo-1-fluoro-3-[(4-methoxyphenyl)methoxy]-2-nitro-benzene (10 g, 28.08 mmol) in ethanol (200 mL) and water (20 mL). The resulting mixture was stirred overnight at 80 °C under a nitrogen atmosphere. LCMS showed that the reaction was complete. The reaction mixture was filtered. The filtrate was concentrated under reduced pressure. The resulting residue was purified by silica gel column chromatography (PE / EA = 9 / 1) to give the desired product 4-bromo-2-fluoro-6-[(4-methoxyphenyl)methoxy]aniline as a pale yellow solid (6 g, 65.50% yield). MS: m / z: C 14 H 13 BrFNO2[M+H] + The calculated value was 326, and the measured value was 326.

[0180] Step 3: Synthesis of tert-butyl (4-bromo-2-fluoro-6-((4-methoxybenzyl)oxy)phenyl)glycinate (1-4)

[0181] At room temperature, K2CO3 (7.49 g, 54.27 mmol) was added to a stirred solution of 4-bromo-2-fluoro-6-[(4-methoxyphenyl)methoxy]aniline (5.9 g, 18.09 mmol) and tert-butyl 2-bromoacetate (10.58 g, 54.27 mmol) in DMF (90 mL). The resulting mixture was stirred at 100 °C for 48 h. LCMS showed that the starting material was completely depleted. The reaction mixture was filtered and the filtrate was washed 3 times with brine. The organic phase was dried over sodium sulfate, filtered and concentrated. The residue was subjected to silica gel column chromatography to give the product as a mixture. The mixture was further purified by reverse-phase flash chromatography (H2O / ACN containing 0.05% NH4HCO3) to give tert-butyl 2-[4-bromo-2-fluoro-6-[(4-methoxyphenyl)methoxy]phenylamino]acetate as a white solid (5 g, 62.70% yield). MS: m / z: C 20 H 23 BrFNO4[M+H] + The calculated value was 440, and the measured value was 440.

[0182] Step 4: Synthesis of N-(4-bromo-2-fluoro-6-((4-methoxybenzyl)oxy)phenyl)-N-sulfamoyl-glycine tert-butyl ester (1-5)

[0183] At 0 °C, a solution of sulfamoyl chloride (2.6 g, 22.48 mmol) in DMA (4 mL) was added to a stirred solution of tert-butyl 2-[4-bromo-2-fluoro-6-[(4-methoxyphenyl)methoxy]phenylamino]acetate (3.3 g, 7.49 mmol) in DMA (80 mL). The reaction mixture was stirred overnight at room temperature. LCMS showed complete depletion of the starting material. The mixture was diluted with ethyl acetate (300 mL) and washed 6 times with brine until all the DMA was removed. The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to give tert-butyl 2-[4-bromo-2-fluoro-6-[(4-methoxyphenyl)methoxy]-N-sulfamoyl-phenylamino]acetate as a brown oil (4 g, 7.70 mmol, 102.70% yield). MS: m / z: C 20 H 24 BrFN2O6S[M-H] - Calculated for is 517, found 517.

[0184] Step 5: Synthesis of 5-(4-bromo-2-fluoro-6-((4-methoxybenzyl)oxy)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide (Int-1)

[0185] At 0 °C, MeOH containing 30% NaOMe (8.32 g, 46.30 mmol) was added to a stirred solution of tert-butyl 2-[4-bromo-2-fluoro-6-[(4-methoxyphenyl)methoxy]-N-sulfamoyl-phenylamino]acetate (4 g, 7.70 mmol) in methanol (20 mL). The mixture was stirred overnight at room temperature. LCMS showed complete depletion of the starting material. The mixture was concentrated. The resulting suspension was dissolved in water (200 mL) and extracted with ethyl acetate. The organic phase was separated and discarded. The aqueous layer was diluted with ethyl acetate, acidified to pH = 3 with 1N HCl solution and extracted 3 times with ethyl acetate. The combined organic layers were washed with brine, dried over sodium sulfate, filtered, and concentrated in vacuo. The resulting residue was further purified by a reverse-phase column (H2O and MeCN containing 0.05% NH4CO3) to give 5-[4-bromo-2-fluoro-6-[(4-methoxyphenyl)methoxy]phenyl]-1,1-dioxo-1,2,5-thiadiazolidine-3-one as an off-white solid (2.50 g, 5.61 mmol, 72.90% yield). MS: m / z: C 16 H 14 BrFN2O5S[M-H] - Calculated for is 443, found 443.

[0186] Step 6: Synthesis of 5-(2-Fluoro-6-((4-methoxybenzyl)oxy)-4-vinylphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide (1-6)

[0187] To a solution of 5-[4-Bromo-2-fluoro-6-[(4-methoxyphenyl)methoxy]phenyl]-1,1-dioxo-1,2,5-thiadiazolidine-3-one (2 g, 4.49 mmol) and tributyl(vinyl)stannane (2.85 g, 8.98 mmol) in DMA (20 mL) was added P(t-Bu)3HBF4 (0.43 g, 0.90 mmol) and Pd2(dba)3 (0.41 g, 0.45 mmol). The resulting mixture was purged with nitrogen for 5 minutes. Subsequently, the mixture was stirred at 80 °C for 12 h. LCMS showed complete depletion of the starting material. The reaction mixture was filtered and the filtrate was purified directly by reverse-phase column to give 5-[2-Fluoro-6-[(4-methoxyphenyl)methoxy]-4-vinylphenyl]-1,1-dioxo-1,2,5-thiadiazolidine-3-one as a pale yellow semi-solid (1.2 g, 3.05 mmol, 68.08% yield). MS: m / z: C 18 H 18 FN2O5S [M-H] - Calculated value was 391, found value was 391.

[0188] Step 7: Synthesis of 4-(1,1-Dioxo-4-oxo-1,2,5-thiadiazolidin-2-yl)-3-fluoro-5-((4-methoxybenzyl)oxy)benzaldehyde (Int-2)

[0189] To a stirred solution of 5-[2-fluoro-6-[(4-methoxyphenyl)methoxy]-4-ethenyl-phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one (970 mg, 2.47 mmol), citric acid (1.04 g, 4.94 mmol) and NMO (579.18 mg, 4.94 mmol) in tert-butanol (6 mL) and water (6 mL) was added K2OsO4 (91.07 mg, 0.25 mmol). The resulting mixture was stirred at room temperature for 1 h. LCMS showed complete conversion of the starting material to the intermediate. Subsequently, NaIO4 (1.07 mL, 7.42 mmol) was added to the mixture 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 and extracted 4 times with ethyl acetate. The organic phase was dried over sodium sulfate, filtered and concentrated. The resulting residue was purified by reverse phase column (0.05% NH4CO3, H2O / ACN) to give 3-fluoro-5-[(4-methoxyphenyl)methoxy]-4-(1,1,4-trioxido-1,2,5-thiadiazolidin-2-yl)benzaldehyde as a brown solid (500 mg, 1.26 mmol, 51.20% yield). MS: m / z: C 17 H 15 FN2O6S[M-H] - Calculated for 393, found 393.

[0190] Preparation Example

[0191] Example 1: 5-[4-[[[4-(2,4-dimethylphenyl)thiazol-2-yl]-methyl-amino]methyl]-2-fluoro-6-hydroxy-phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one

[0192]

[0193] Scheme 1:

[0194]

[0195] Step 1: Add acetic acid (91.36 mg, 1.52 mmol) to a stirred solution of methylamine (70.88 mg, 2.28 mmol) and 3-fluoro-5-[(4-methoxyphenyl)methoxy]-4-(1,1,4-trioxo-1,2,5-thiadiazolidin-2-yl)benzaldehyde (Int-2, 300 mg, 0.76 mmol) in methanol (10 mL). Stir the mixture at room temperature for 1 h. Subsequently, add NaBH3CN (191.09 mg, 3.04 mmol) to the reaction mixture at 0 °C. Stir the resulting mixture at room temperature for an additional 1 h. After monitoring the completion of the reaction by LCMS, concentrate the mixture. The resulting residue was purified by reverse-phase column chromatography to give 5-[2-fluoro-6-[(4-methoxyphenyl)methoxy]-4-(methylaminomethyl)phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one as a white solid (210 mg, 0.51 mmol, 67.42% yield). MS: m / z: C 18 H 20 FN3O5S [M+H] + The calculated value is 410, and the measured value is 410.

[0196] Step 2: Add DIEA (0.02 mL, 1.5 mmol) to a stirred solution of 5-[2-fluoro-6-[(4-methoxyphenyl)methoxy]-4-(methylaminomethyl)phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one (205 mg, 0.50 mmol) and 4-bromo-2-chloro-thiazole (198.75 mg, 1 mmol) in DMSO (4 mL). Stir the mixture solution at 80 °C for 16 h. LCMS showed complete depletion of the starting material. The mixture was purified by reverse-phase column chromatography to give 5-[4-[[(4-bromothiazol-2-yl)-methyl-amino]methyl]-2-fluoro-6-[(4-methoxyphenyl)methoxy]phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one as a brown solid (120 mg, 0.21 mmol, 41.94% yield). MS: m / z: C 21 H 20 BrFN4O5S2 [M+H] + The calculated value is 571, and the measured value is 571.

[0197] Step 3: Under nitrogen, Pd(dppf)Cl2 (38.59 mg, 0.05 mmol) was added to a stirred solution of 5-[4-[[(4-bromothiazol-2-yl)-methyl-amino]methyl]-2-fluoro-6-[(4-methoxyphenyl)methoxy]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one (90 mg, 0.16 mmol), (2,4-dimethylphenyl)boronic acid (35.43 mg, 0.24 mmol) and Na2CO3 (50.08 mg, 0.47 mmol) in 1,4-dioxane (10 mL) and water (1 mL). The resulting mixture was stirred at 80 °C for 2 h. After completion of the reaction monitored by LCMS, the mixture was concentrated. The resulting residue was purified by reverse-phase column to give 5-[4-[[[4-(2,4-dimethylphenyl)thiazol-2-yl]-methyl-amino]methyl]-2-fluoro-6-[(4-methoxyphenyl)methoxy]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one as a yellow solid (60 mg, 0.10 mmol, 63.84% yield). MS: m / z: C 29 H 29 FN4O5S2[M+H] + Calculated value for is 597, found value is 597.

[0198] Step 4: TFA (3 mL) was added to a stirred solution of 5-[4-[[[4-(2,4-dimethylphenyl)thiazol-2-yl]-methyl-amino]methyl]-2-fluoro-6-[(4-methoxyphenyl)methoxy]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one (60 mg, 0.10 mmol) in DCM (3 mL), and the mixture was stirred at room temperature for 2 h. After completion, the reaction mixture was concentrated. The resulting residue was purified by reverse-phase flash chromatography (H2O and ACN containing 0.05% NH4HCO3) and further purified by preparative HPLC to give 5-[4-[[[6-(dimethylamino)-3-pyridinyl]amino]methyl]-2-fluoro-6-hydroxy-phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one as a green solid (3 mg, 5.69% yield). MS: m / z: C 21 H 21 FN4O4S2,[M+H] + Calculated value for is 477; found value is 477. 11H NMR (400 MHz, DMSO-d6) δ 10.36 (s, 1H), 7.47 (d, J = 7.8 Hz, 1H), 7.06 - 6.95 (m, 2H), 6.79 (s, 1H), 6.72 - 6.64 (m, 2H), 4.67 (s, 2H), 4.31 (s, 2H), 3.09 (s, 3H), 2.36 (s, 3H), 2.28 (s, 3H).

[0199] Preparative HPLC purification conditions: Column: Welch Utimate AQ C18, 50 * 250 mm, 10 μm; Mobile phase A: water (0.1% FA), Mobile phase B: ACN; Flow rate: 100 mL / min; Gradient: within 20 min, 40% B to 65% B, 65% B; Wavelength: 254 nm.

[0200] Example 2: 5-[2-Fluoro-6-hydroxy-4-[(1H-pyrazol-3-ylamino)methyl]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one

[0201]

[0202] According to the preparation of Example 4, the title compound in the form of a white solid was obtained in an overall yield of 8.78% using 1H-pyrazol-3-amine in Step 1. MS: m / z: C 12 H 12 FN5O4S, [M + H] + The calculated value is 342; the measured value is 342. 1 1H NMR (400 MHz, DMSO-d6) δ 9.39 (s, 1H), 7.65 (s, 1H), 6.60 - 6.50 (m, 3H), 3.96 (s, 2H), 3.62 (s, 2H).

[0203] 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: within 6.5 min, 20% B to 40% B, 40% B; Wavelength: 254 / 210 nm.

[0204] Example 3: 5-[2-Fluoro-6-hydroxy-4-[(isoxazol-3-ylamino)methyl]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one

[0205]

[0206] Prepared according to Example 4, the title compound as a white solid was obtained in 4.75% overall yield using isoxazol-3-amine in Step 1. MS: m / z: C 12 H 11 FN4O5S, [M+H] + Calculated for 343; found 343. 1 H NMR (400 MHz, DMSO-d6) δ 8.35 (d, J = 1.8 Hz, 1H), 6.71 (d, J = 1.9 Hz, 1H), 6.66 (dd, J = 10.7, 1.8 Hz, 1H), 6.00 (d, J = 1.8 Hz, 1H), 4.20 (d, J = 10.8 Hz, 4H).

[0207] 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 in 6.5 min, 60% B; Wavelength: 254 / 210 nm.

[0208] Example 4: 5-[2-Fluoro-6-hydroxy-4-[[(5-methylisoxazol-3-yl)amino]methyl]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one

[0209]

[0210] Scheme 2:

[0211]

[0212] At 0 °C, TMSOTf (84.44 mg, 0.38 mmol) was added to a stirred solution of 3-fluoro-5-[(4-methoxyphenyl)methoxy]-4-(1,1,4-trioxo-1,2,5-thiadiazolidin-2-yl)benzaldehyde (100 mg, 0.25 mmol) and 5-methylisoxazol-3-amine (61.66 mg, 0.38 mmol) in DCM (8 mL). The reaction mixture was stirred at room temperature for 2 h. The mixture was cooled to 0 °C, and NaBH(AcO)3 (107.51 mg, 0.51 mmol) was slowly added to the above mixture. After the addition, the resulting mixture was stirred at room temperature for another 16 h. LCMS showed complete depletion of the starting material (about 50% of the desired product and 10% of the PMB-protected intermediate were observed). TFA (10 mL) was added to the reaction mixture at 0 °C. The resulting mixture was stirred at room temperature for another 3 h. LCMS showed complete cleavage of the PMB protecting group, and the reaction mixture was concentrated. The residue was purified by reverse-phase column chromatography (H2O and MeCN containing 0.05% NH4HCO3) and further purified by preparative HPLC to give 5-[2-fluoro-6-hydroxy-4-[[(5-methylisoxazol-3-yl)amino]methyl]phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one as a yellow solid (9 mg, 8.02% yield). MS: m / z: C 13 H 13 FN4O5S, [M+H] + The calculated value is 357; the measured value is 357. 1 1H NMR (400 MHz, DMSO-d6) δ 7.60 (s, 2H), 6.65 (s, 1H), 6.64 - 6.52 (m, 2H), 5.66 (s, 1H), 4.13 (d, J = 6.2 Hz, 2H), 3.94 (d, J = 2.2 Hz, 2H), 2.21 (s, 3H).

[0213] Preparative HPLC purification conditions: Column: XBridge preparative phenyl OBD column, 19*100 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3), Mobile phase B: ACN; Flow rate: 50 mL / min; Gradient: within 8 min, 35% B to 45% B, 45% B; Wavelength: 254 / 210 nm.

[0214] Example 5: 5-[2-Fluoro-6-hydroxy-4-[(1H-1,2,4-triazol-3-ylamino)methyl]phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one

[0215]

[0216] Process 3:

[0217]

[0218] Step 1: At 0 °C, dropwise add TMSCl (0.08 mL, 0.62 mmol) to a stirred solution of 3-fluoro-5-[(4-methoxyphenyl)methoxy]-4-(1,1,4-trioxo-1,2,5-thiadiazolidin-2-yl)benzaldehyde (Int-2, 100 mg, 0.25 mmol) and 1H-1,2,4-triazol-3-amine (34.2 mg, 0.28 mmol) in anhydrous DMF (6 mL). Then stir the resulting mixture at room temperature for 30 min. Subsequently, cool the reaction mixture to 60 °C and slowly add a solution of BH3 in THF (1 M, 0.45 mL, 0.46 mmol) using a syringe. After the addition, stir the reaction mixture at room temperature for 1 h. LCMS shows the reaction is complete. Quench the resulting solution with ice water (1 mL) and purify directly by reverse-phase column (H2O and MeCN containing 0.05% NH4HCO3) to obtain 5-[2-fluoro-6-hydroxy-4-[(1H-1,2,4-triazol-3-ylamino)methyl]phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one 5-(4-(((2,6-dimethylpyridin-4-yl)amino)methyl)-2-fluoro-6-((4-methoxybenzyl)oxy)phenyl)-1,2,5-thiadiazolidin-3-one 1,1-dioxide as a yellow solid (76 mg, 0.14 mmol, 56.40% yield). MS: m / z: C 24 H 20 ClFN4O5S [M+H] + Calculated value for is 531; found value is 531.

[0219] Step 2: Prepared according to the preparation of Example 1, the title compound as a white solid was obtained in 5.99% yield using 3-1 in Step 2.

[0220] MS: m / z: C 11 H 11 FN6O4S, [M+H] + Calculated value for is 343; found value is 343. 1 1H NMR (400 MHz, DMSO-d6 + D2O) δ 6.86 - 6.50 (m, 2H), 4.23 (s, 2H), 3.94 (s, 2H).

[0221] Preparative HPLC purification conditions: Column: XBridge BEH C18 OBD preparative column, 19*250 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3), Mobile phase B: ACN; Flow rate: 25 mL / min; Gradient: within 6 min, 8% B to 20% B, 20% B; Wavelength: 254 nm.

[0222] Example 6: 5-[2-Fluoro-6-hydroxy-4-[[(1-methylpyrazol-3-yl)amino]methyl]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one

[0223]

[0224] According to the preparation of Example 4, 1-methylpyrazol-3-amine was used in Step 1 to obtain the title compound as a white solid in an overall yield of 6.93%. MS: m / z: C 13 H 14 FN5O4S, [M+H] + The calculated value is 356; the measured value is 356. 1 H NMR (400 MHz, DMSO-d6 + D2O) δ 7.52 - 7.51 (m, 1H), 6.72 - 6.59 (m, 2H), 4.17 (s, 2H), 4.09 (s, 2H), 3.62 (d, J = 1.6 Hz, 3H).

[0225] 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: within 5.5 min, 15% B to 28% B, 28% B; Wavelength: 210 / 254 nm.

[0226] Example 7: 5-[2-Fluoro-6-hydroxy-4-[(1,3,4-thiadiazol-2-ylamino)methyl]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one

[0227]

[0228] According to the preparation of Example 5, 1,3,4-thiadiazol-2-amine was used in Step 1 to obtain the title compound as a white solid in an overall yield of 8.99%. MS: m / z: C 11 H 10 FN5O4S2, [M+H] + The calculated value is 360; the measured value is 360. 11H NMR (400 MHz, DMSO-d6) δ 8.64 (s, 1H), 8.27 (t, J = 5.8 Hz, 1H), 6.87 - 6.52 (m, 2H), 4.39 (d, J = 5.8 Hz, 2H), 3.93 (s, 2H).

[0229] Preparative HPLC purification conditions: Column: XBridge BEH C18 OBD preparative column, 19 * 250 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3), Mobile phase B: ACN; Flow rate: 25 mL / min; Gradient: within 7 min, 10% B to 25% B, 25% B; Wavelength: 254 nm.

[0230] Example 8: 5-[2-Fluoro-6-hydroxy-4-[[(5-methyl-1H-imidazol-2-yl)amino]methyl]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one

[0231]

[0232] According to the preparation of Example 5, 5-methyl-1H-imidazol-2-amine hydrochloride was used in Step 1 to obtain the title compound as a white solid in an overall yield of 7.52%. MS: m / z: C 13 H 14 FN5O4S, [M + H] + Calculated value is 356; Measured value is 356. 1 1H NMR (400 MHz, DMSO-d6) δ 11.75 (s, 1H), 9.45 (s, 1H), 7.20 (s, 2H), 6.56 (d, J = 9.5 Hz, 2H), 3.99 (d, J = 25.2 Hz, 2H), 3.70 (s, 2H), 2.11 (s, 3H).

[0233] 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: 60 mL / min; Gradient: within 7 min, 22% B to 40% B, 45% B; Wavelength: 254 / 210 nm.

[0234] Example 9: 5-[2-Fluoro-6-hydroxy-4-[(thiazol-2-ylamino)methyl]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one

[0235]

[0236] Process 4:

[0237]

[0238] Step 1: At 0 °C, Ti(i-PrO)4 (172.83 mg, 0.61 mmol) was added to a stirred solution of 3-fluoro-5-[(4-methoxyphenyl)methoxy]-4-(1,1,4-trioxo-1,2,5-thiadiazolidin-2-yl)benzaldehyde (60 mg, 0.15 mmol) and thiazol-2-amine (36 mg, 0.36 mmol) in DCE (10 mL). The resulting mixture was stirred at 60 °C for 12 h. At 0 °C, NaBH3CN (19.47, 0.30 mmol) was added to the mixture. The resulting mixture was stirred at room temperature for an additional 2 h. After completion, the reaction mixture was concentrated. The resulting residue was dissolved in DMSO and purified directly by a reverse-phase column (H2O and MeCN containing 0.05% TFA) to give 5-[2-fluoro-6-[(4-methoxyphenyl)methoxy]-4-[(thiazol-2-ylamino)methyl]phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one as a white solid (70.0 mg, 0.14 mmol, 96.15% yield). MS: m / z: C 20 H 19 FN4O5S2[M+H] + The calculated value is 479; the measured value is 479.

[0239] Step 2: At room temperature, TFA (2 mL) was added to a stirred solution of 5-[2-fluoro-6-[(4-methoxyphenyl)methoxy]-4-[(thiazol-2-ylamino)methyl]phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one (70 mg, 0.15 mmol) in DCM (6 mL). The resulting mixture was stirred at room temperature for 12 h. After completion, the reaction mixture was concentrated. The resulting residue was purified by reverse-phase column chromatography (H2O and MeCN containing 0.05% TFA) and further purified by preparative HPLC to give 5-[2-fluoro-6-hydroxy-4-[(thiazol-2-ylamino)methyl]phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one as a brown solid (5.7 mg, 0.01 mmol, 10.64% yield). MS: m / z: C 12 H 11 FN4O4S2[M+H] + The calculated value is 359, and the measured value is 359. 11H NMR (400 MHz, DMSO-d6) δ 7.13 (t, J = 3.6 Hz, 1H), 6.79 - 6.74 (m, 1H), 6.66 (d, J = 12.0 Hz, 2H), 4.40 (s, 2H), 4.01 (d, J = 2.2 Hz, 2H).

[0240] Preparative HPLC purification conditions: Column: XBridge Shield RP18 OBD column, 30 * 150 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: within 8 min, 5% B to 35% B, 35% B; Wavelength: 254 / 210 nm.

[0241] Example 10: 5-[2-Fluoro-6-hydroxy-4-[(1H-imidazol-2-ylamino)methyl]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one

[0242]

[0243] According to the preparation of Example 5, 1H-imidazol-2-amine was used in Step 1 to obtain the title compound as a white solid in an overall yield of 8.56%. MS: m / z: C 12 H 12 FN5O4S, [M + H] + Calculated value is 342; Observed value is 342. 1H NMR (400 MHz, DMSO-d6 + D2O) δ 6.86 (s, 2H), 6.70 (d, J = 8.4 Hz, 2H), 4.38 (s, 2H), 4.06 (s, 2H).

[0244] 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: within 6.59 min, 10% B to 30% B, 30% B; Wavelength: 254 / 210 nm.

[0245] Example 11: 5-[2-Fluoro-6-hydroxy-4-[[(1-methylimidazol-2-yl)amino]methyl]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one

[0246]

[0247] Process 5:

[0248]

[0249] Step 1: At 0 °C, a solution of titanium tetraisopropoxide (115.63 mg, 0.41 mmol) was added to a solution of 3-fluoro-5-[(4-methoxyphenyl)methoxy]-4-(1,1,4-trioxo-1,2,5-thiadiazolidin-2-yl)benzaldehyde (80 mg, 0.2 mmol) and 1-methylimidazol-2-amine (39.4 mg, 0.41 mmol) in dichloromethane (8 mL). The reaction mixture was stirred at room temperature for 2 h. At 0 °C, NaBH(AcO)3 (38.95 mg, 0.41 mmol) was added to the mixture. The resulting mixture was stirred at room temperature for an additional 30 min. After completion, the reaction mixture was concentrated. The resulting residue was dissolved in DMSO and purified by reverse-phase column (H2O and MeCN containing 0.05% NH4HCO3) to give 5-[2-fluoro-6-[(4-methoxyphenyl)methoxy]-4-[[(1-methylimidazol-2-yl)amino]methyl]phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one (60 mg, 62.5% yield). MS: m / z: C 21 H 22 FN5O5S, [M+H] + Calculated value for is 476; found value is 476.

[0250] Step 2: According to the preparation of Example 1, the title compound in white solid form was prepared in 30.50% yield using 16-1 in Step 2. MS: m / z: C 13 H 14 FN5O4S, [M+H] + Calculated value for is 356; found value is 356. 1 H NMR (400 MHz, DMSO-d6) δ 12.40 (s, 1H), 9.57 (s, 1H), 8.43 (t, J = 6.1 Hz, 1H), 7.10 (d, J = 2.4 Hz, 1H), 7.05 (d, J = 2.4 Hz, 1H), 6.73 (d, J = 11.1 Hz, 2H), 4.45 (d, J = 6.1 Hz, 2H), 3.98 (s, 2H), 3.54 (s, 3H).

[0251] 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: 23% B to 45% B in 5 min, 45% B; Wavelength: 210 / 254 nm.

[0252] Example 12: 5-[2-Fluoro-6-hydroxy-4-[[(5-methyl-1,3,4-thiadiazol-2-yl)amino]methyl]phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one

[0253]

[0254] Procedure 6:

[0255]

[0256] Step 1: Titanium(IV) isopropoxide (65.93 mg, 0.25 mmol) was added dropwise to a stirred mixture of 3-fluoro-5-[(4-methoxyphenyl)methoxy]-4-(1,1,4-trioxo-1,2,5-thiadiazolidin-2-yl)benzaldehyde (50 mg, 0.13 mmol) and 5-methyl-1,3,4-thiadiazol-2-amine (14.6 mg, 0.13 mmol) in DCE (5 mL). The resulting mixture was stirred at room temperature for 12 h. At 0 °C, sodium cyanoborohydride (23.86, 0.38 mmol) was added to the above mixture. The resulting mixture was stirred at room temperature for an additional 1 h. The reaction mixture was purified directly by reverse-phase column chromatography (H2O and MeCN containing 0.05% TFA) to afford 5-[2-fluoro-6-[(4-methoxyphenyl)methoxy]-4-[[(5-methyl-1,3,4-thiadiazol-2-yl)amino]methyl]phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one as an off-white oil (50 mg TFA salt, 0.10 mmol, 79.90% yield). MS: m / z: C 20 H 20 FN5O5S2 [M+H] + Calculated for 494; found 494.

[0257] Step 2: Trifluoroacetic acid (4 mL) was added to a stirred mixture of 5-[2-fluoro-6-[(4-methoxyphenyl)methoxy]-4-[(1,3,4-thiadiazol-2-ylamino)methyl]phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one (45 mg, 0.09 mmol) in DCM (4 mL) at room temperature. The resulting mixture was stirred at room temperature for 12 h. After completion of the reaction was monitored by LCMS, the mixture was concentrated under reduced pressure. The resulting residue was purified by preparative HPLC to afford 5-[2-fluoro-6-hydroxy-4-[[(5-methyl-1,3,4-thiadiazol-2-yl)amino]methyl]phenyl]-1,1-dioxo-1,2,5-thiadiazolidin-3-one as a white solid (6.6 mg, 0.01 mmol, 18.17% yield). MS: m / z: C12 H 12 FN5O4S2[M+H] + The calculated value is 374; the measured value is 374. 1 H NMR (400 MHz, DMSO-d6 + D2O) δ 7.58 - 7.50 (m, 1H), 6.82 - 6.48 (m, 5H), 4.38 (s, 2H), 3.94 (s, 2H).

[0258] Preparative HPLC purification conditions: Column: XBridge Preparative OBD C18 column, 19 * 250 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3), Mobile phase B: ACN; Flow rate: 25 mL / min; Gradient: within 6 min, 28% B to 48% B, 48% B; Wavelength: 254 / 210 nm.

[0259] Example 13: 5-[2-Fluoro-6-hydroxy-4-[[(4-methylthiazol-2-yl)amino]methyl]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one

[0260]

[0261] According to the preparation of Example 9, the title compound as a white solid was obtained in a total yield of 14.16% using 4-methylthiazol-2-amine in Step 1. After adding NaBH3CN, the reaction was carried out at 80 °C instead of room temperature. MS: m / z: C 13 H 13 FN4O4S2,[M+H] + The calculated value is 373; the measured value is 373. 1H NMR (400 MHz, DMSO-d6) δ 6.71 - 6.58 (m, 2H), 6.22 (d, J = 1.3 Hz, 1H), 4.32 (s, 2H), 3.99 (s, 2H), 2.07 (s, 3H).

[0262] Preparative HPLC purification conditions: Column: XBridge Shield RP18 OBD column, 30 * 150 mm, 5 μm; Mobile phase A: water (10 mmol / L NH4HCO3), Mobile phase B: ACN; Flow rate: 60 mL / min; Gradient: within 8 min, 55% B to 75% B, 75% B; Wavelength: 254 / 210 nm.

[0263] Example 14: 5-[2-Fluoro-6-[(4-methoxyphenyl)methoxy]-4-[[(3-methylisothiazol-5-yl)amino]methyl]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one

[0264]

[0265] Prepared according to Example 4, using 3-methylisothiazol-5-amine in Step 1, the title compound was obtained as a white solid in a total yield of 7.31%. MS: m / z: C 13 H 13 FN4O4S2, [M+H] + Calculated value is 373; found value is 373. 1 1H NMR (400 MHz, DMSO-d6) δ 10.37 (s, 1H), 7.80 (s, 1H), 6.78 - 6.62 (m, 2H), 6.03 (s, 1H), 4.34 (s, 2H), 4.23 (s, 2H), 2.16 (s, 3H).

[0266] Purification conditions: The compound was purified by reverse-phase column chromatography (H2O and MeCN containing 0.05% NH4HCO3).

[0267] Example 15: 5-[2-Fluoro-6-hydroxy-4-[[(3-methyl-1H-pyrazol-5-yl)amino]methyl]phenyl]-1,1-dioxido-1,2,5-thiadiazolidin-3-one

[0268]

[0269] Prepared according to Example 9, using 5-methyl-1H-pyrazol-3-amine in Step 1, the title compound was obtained as a white solid in a total yield of 12.20%. MS: m / z: C 13 H 14 FN5O4S, [M+H] + Calculated value is 356; found value is 356. 1 1H NMR (400 MHz, DMSO-d6) δ 13.8 - 12.8 (m, 1H), 9.60 (s, 1H), 7.47 (s, 1H), 6.63 (d, J = 12.5 Hz, 2H), 5.60 (s, 1H), 4.22 (s, 2H), 3.99 (s, 2H), 2.19 (s, 3H).

[0270] Purification conditions for preparative HPLC: 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: from 40% B to 60% B in 6.58 min, 60% B; Wavelength: 254 / 210 nm.

[0271] Examples of the compounds prepared by the above procedures are listed in Table 1.

[0272] Examples of the Prepared Compounds

[0273] Table 1

[0274]

[0275]

[0276]

[0277] Bioanalysis

[0278] The pharmacological properties of the compounds of the present invention can be confirmed by various bioassays known in the art. The compounds of the present invention have been subjected to the bioassays exemplified below.

[0279] Assay

[0280] Performed as described by the vendor (AssayQuant Technologies, Marlborough, MA) Kinase assay. Briefly, in a 384-well reagent plate, serial dilutions of a 10 mM DMSO stock solution were made at 3-fold intervals to prepare 1000X compound solutions in DMSO. Then, 50 nL of the compound dilution series was added to the corresponding wells of a 384-well assay plate. 40 μL of 1X 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 × substrate (AQT0264) was transferred to each well of the assay plate to achieve a final substrate concentration of 20 μM. Finally, 10 μL of a 5X PTPN2 enzyme stock solution was added to each well of the assay plate, with a final enzyme concentration of 150 pM. The reaction progress curve was collected by sampling the fluorescence intensity every 71 seconds for one hour at room temperature using a Synergy H4 plate reader (BioTek Instruments / Agilent Technologies, Winooki, VT) at an excitation wavelength of 360 nm (λ ex 360) and an emission wavelength of 480 nm (λ em 480).

[0281] Phosphatase activity assay using DIFMUP as the substrate:

[0282] The PTPN2 biochemical analysis was performed as follows: A 5× human PTPN2 (SRP5075, MilliporeSigma, Burlington, MA) stock solution and a 1.25× DiFMUP (D6567, ThermoFisher Scientific, Waltham, MA) stock solution were prepared in 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 μL of the DiFMUP substrate solution (final concentration of 25 mM DiFMUP substrate) was added to a Corning 3574 384-well white non-binding surface microtiter plate containing 0.05 μL of serially diluted test compounds prepared in DMSO. The reaction was initiated by adding 10 μL of the enzyme solution to a final PTPN2 concentration of 0.15 nM, and then monitored every 105 seconds at room temperature in a BioTek Synergy HTX plate reader (Agilent Technologies, Santa Clara, CA) at λ EX 360 / λ EM 460 for 60 minutes. The initial linear portion of the progress curve was fit to a linear equation to obtain the slope and converted to % inhibition based on the 100% activity value of the uninhibited control. The IC 50 value of each compound was obtained by fitting the % inhibition-compound concentration curve using Dotmatics software (Dotmatics, Bishops Stortford, Hertfordshire, England).

[0283] Cell Proliferation Assay Protocol

[0284] 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 a total volume of 20 μL into two white opaque 384-well tissue culture-treated microplates (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 handler (Beckman Coulter, Indianapolis, IN). Negative control wells received only 30 nL of DMSO (0.15% final concentration). The microplates were returned to the incubator for 1 h and then the cells were treated with 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 pipetting platform (INTEGRA Biosciences, Hudson, NH). The microplates were incubated at 37 °C for 4 days and then cell proliferation was analyzed using CellTiter-Glo reagent (Promega, Madison, WI, #G7573, 25 μL / well). Luminescence signal intensity was collected 15 min after the addition of CellTiter-Glo reagent using an EnVision 2105 plate reader (PerkinElmer) and analyzed using the Dotmatics software platform to calculate the IC 50 value. Off-target compound-mediated cytotoxicity was identified by examining growth inhibition in the absence of IFNγ.

[0285] Phospho-STAT1 assay protocol

[0286] 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 10,000 cells / well in a total volume of 20 μL into white opaque 384-well tissue culture-treated microplates (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 handler (Beckman Coulter, Indianapolis, IN). Negative control wells received only 30 nL of DMSO (0.15% final concentration). The plates were returned to the incubator for 1 hour, then the cells were treated with 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 pipetting platform (INTEGRA Biosciences, Hudson, NH). The plates were incubated at 37 °C for 1 hour and the phosphorylated STAT1 protein levels were analyzed using a phosphorylated-STAT1 (Tyr701) HTRF kit (Cisbio, Bedford, MA, #63ADK026PEH) according to the manufacturer's instructions. After 24 hours, the HTRF signal intensity was collected using an EnVision 2105 plate reader (PerkinElmer) and analyzed using the Dotmatics software platform to calculate the IC 50 value.

[0287] Bioanalytical data

[0288] Table 2 is an overview of the bioanalytical data for the prepared examples / embodiments. For IC 50 data, high DDT concentration and / or DiFMUP substrate assays were used; those skilled in the art can use either assay. Columns or rows with double asterisks indicate that an IC 50 value or embodiment has been provided.

[0289] Table 2

[0290]

[0291]

[0292]

Claims

1. A compound having the structure of formula I: Among them, Independently at each occurrence: R 1 selected from 1H-1,2,4-triazol-3-yl, R 2 selected from -H, alkyl, and substituted alkyl; R 3 selected from -H, alkyl, and substituted alkyl; R 4 selected from -H, alkyl, substituted alkyl, branched alkyl, alkoxy, halogen, cyano, amine, hydroxy, aryl, and substituted aryl; R 5 selected from -H, alkyl, substituted alkyl, branched alkyl, alkoxy, halogen, cyano, amine, hydroxy, aryl and substituted aryl.

2. The compound according to claim 1, wherein: R 1 selected from R 3 selected from -H and -CH3; R 4 are -H and -CH3.

3. The compound according to claim 1, wherein: R 1 For R 4 Selected from -H and -CH3.

4. The compound according to claim 1, wherein: R 1 For R 4 selected from -H, -CH3 and 2,4-dimethylphenyl; R 5 selected from -H and -CH3.

5. The compound according to claim 1, wherein: R 1 selected from R 4 Selected from -H and -CH3.

6. The compound according to claim 1, wherein: R 1 For R 3 selected from -H and -CH3; R 4 are -H and -CH3; R 5 are -H and -CH3.

7. A compound selected from: 5-(4-(((4-(2,4-dimethylphenyl)thiazol-2-yl)(methyl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(4-(((1H-pyrazol-3-yl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(2-fluoro-6-hydroxy-4-((isoxazol-3-ylamino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(2-fluoro-6-hydroxy-4-(((5-methylisoxazol-3-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(4-(((1H-1,2,4-triazol-3-yl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(2-fluoro-6-hydroxy-4-(((1-methyl-1H-pyrazol-3-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(4-(((1,3,4-thiadiazol-2-yl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(2-fluoro-6-hydroxy-4-(((5-methyl-1H-imidazol-2-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(2-fluoro-6-hydroxy-4-((thiazol-2-ylamino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(4-(((1H-imidazol-2-yl)amino)methyl)-2-fluoro-6-hydroxyphenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(2-fluoro-6-hydroxy-4-(((1-methyl-1H-imidazol-2-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(2-fluoro-6-hydroxy-4-(((5-methyl-1,3,4-thiadiazol-2-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(2-fluoro-6-hydroxy-4-(((4-methylthiazol-2-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(2-fluoro-6-hydroxy-4-(((3-methylisothiazol-5-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide; 5-(2-Fluoro-6-hydroxy-4-(((3-methyl-1H-pyrazol-5-yl)amino)methyl)phenyl)-1,2,5-thiadiazolidine-3-one 1,1-dioxide.

8. A pharmaceutical composition comprising the compound of formula (I) according to claim 1 or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable carrier.

9. A method of treating cancer, comprising administering to the patient a therapeutically effective amount of the compound of formula (I) according to claim 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, gastric cancer, brain cancer, head and neck cancer, skin cancer, uterine cancer, testicular cancer, glioma, esophageal cancer, liver cancer, including hepatoma, lymphoma, including B-acute lymphoblastic lymphoma, non-Hodgkin's lymphoma, Burkitt's lymphoma, small lymphoma, Hodgkin's lymphoma, leukemia, and multiple myeloma.

10. A method of treating cancer in a patient in need thereof, comprising administering to the patient an effective amount of a combination of the compound of claim 1 and an additional therapeutic agent.

11. The method according to claim 10, wherein the additional therapeutic agent is an immunotherapeutic agent.

12. The method according to claim 11, wherein the immunotherapeutic agent is selected from anti-PD-1 antibody, anti-PD-L1 antibody, and anti-CTLA-4 antibody.

13. A method of treating cancer in a patient in need thereof, the method comprising administering to the patient an effective amount of the pharmaceutically acceptable composition according to claim 1.

14. The method according to claim 1, wherein the method of treating cancer is selected from radiation, surgery, chemotherapy, or administration of a biopharmaceutical.

15. The method according to claim 14, wherein the method of treating cancer further comprises administering a biopharmaceutical, wherein the biopharmaceutical is a drug that stimulates the immune system.

16. The method according to claim 15, wherein the method further comprises administering to the individual a DGKα and / or DGKζ inhibitor, an antagonist of the PD1 / PD-L1 axis, and an antagonist of CTLA4.

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