Protein tyrosine phosphatase inhibitors

Abstract

The invention provides a compound with PTPN2 inhibitory activity, a preparation method of the compound and application of the compound in preparation of drugs for treating related diseases. Specifically disclosed are compounds represented by formula (I) and pharmaceutically acceptable salts thereof. # imgabs0 #

Description

[0001] This application claims the priority of the Chinese patent application filed with the China Patent Office on July 20, 2023, with application number 202310897417.1 and invention name “Protein Tyrosine Phosphatase Inhibitors”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The present application belongs to the field of pharmaceutical chemistry and relates to a PTPN2 inhibitor, a preparation method thereof and its application in the preparation of drugs for treating related diseases, and specifically to a compound represented by formula (I) and a pharmaceutically acceptable salt thereof. Background Art

[0003] Protein tyrosine phosphatase 2 (PTPN2), also known as T-cell protein tyrosine phosphatase (TC-PTP), is a member of the non-receptor tyrosine phosphorus-specific phosphatase family that controls the regulatory processes of various cells by removing phosphate groups from tyrosine substrates. It was first cloned from a cDNA library of human peripheral blood T cells in 1989, and subsequent studies have found that PTPN2 is widely expressed in various tissues, but is most highly expressed in hematopoietic and placental cells (Mosinger, B.Jr. et al., Proc Natl Acad Sci USA 89:499–503; 1992). In humans, the expression of PTPN2 is posttranscriptionally 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 junction and a 48 kDa canonical form with a C-terminal ER retention motif (Tillmann U. et al., Mol Cell Biol 14:3030–3040; 1994). Under certain conditions of cellular stress, the 45 kDa isoform can be passively infused into the cytosol. Both isoforms share an N-terminal phosphotyrosine phosphatase catalytic domain. PTPN2 regulates signaling of non-receptor tyrosine kinases (e.g., JAK1, JAK3), receptor tyrosine kinases (e.g., INSR, EGFR, CSF1R, PDGFR), transcription factors (e.g., STAT1, STAT3, STAT5a / b), and Src family kinases (e.g., Fyn, Lck). As a key negative regulator of the JAK-STAT pathway, PTPN2 directly regulates signaling through cytokine receptors, including IFNγ.

[0004] The tumor suppressor effect of PTPN2 began to be reported in 2010. First, Kleppe et al. analyzed samples of patients with T-cell acute lymphoblastic leukemia (T-ALL) and found that 6% of patients had PTPN2 expression loss. Knocking out PTPN2 in cell lines can significantly promote tumor cell proliferation, suggesting that PTPN2 is a tumor suppressor gene in T-ALL. Subsequent studies have found that PTPN2 can negatively regulate the JAK / STAT signaling pathway, which promotes tumor proliferation in many cases, so PTPN2 is considered to be a tumor suppressor gene. However, some studies have found that PTPN2 acts as an oncogene. In Myc-induced mouse B-cell lymphoma, overexpressed PTPN2 can promote cell proliferation, while knocking down PTPN2 significantly inhibits cell proliferation, and even completely inhibits B-cell lymphoma in mice. The inflammatory response in the rectum of mice with conditional knockout of PTPN2 was more severe than that of wild-type mice, but in the colorectal cancer model, it was found that the occurrence and development of colorectal cancer in PTPN2 knockout mice was significantly slower than that in wild-type mice. Some articles have reported that the expression level of PTPN2 in patients with glioma and glioblastoma has a strong correlation with the patient's prognosis. Patients with high expression of PTPN2 often have a poor prognosis, which also suggests that PTPN2 may promote tumors. Summary of the invention

[0005] The purpose of the present application is to provide a compound having PTPN2 inhibitory activity, a stereoisomer thereof or a pharmaceutically acceptable salt thereof.

[0006] In a first aspect, the present application provides a compound of formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof,

[0007]

[0008] in,

[0009] Q is a 5-7 membered carbon ring;

[0010] R2 and R3 are each independently hydrogen, C 1-6 Alkyl; or

[0011] R2 and R3 are combined to form an alkylene group, which together with the carbon atom to which they are attached form a 3- to 4-membered ring and form a condensed ring with ring Q; or

[0012] R2 and R3 are combined to form an alkylene group, which together with the carbon atom to which they are attached form a 3- to 4-membered ring and form a spiro ring with ring Q;

[0013] R4 is -NHR0,

[0014] R0 is selected from C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted by the following groups:

[0015]

[0016] R a , R b , R c and R d are independently selected from H, D, C 1-6 Alkyl, halogen, C 1-6 Haloalkyl;

[0017] R e Selected from H, D, C 1-6 Alkyl, C 1-6 Haloalkyl;

[0018] X is selected from O, S, CH2 or a single bond;

[0019] n is an integer selected from 0, 1, and 2.

[0020] In some embodiments of the present application, Q is a 6-membered carbon ring.

[0021] In some embodiments of the present application, R2 and R3 are each independently hydrogen.

[0022] In some embodiments of the present application, R2 and R3 together with the carbon atoms to which they are attached form a 3- to 4-membered ring and form a condensed ring with ring Q, wherein the 3- to 4-membered ring is a 3- to 4-membered carbon ring.

[0023] In some embodiments of the present application, R2 and R3 together with the carbon atom to which they are attached form a 3-membered ring and form a condensed ring with the ring Q, and the 3-membered ring is a 3-membered carbon ring.

[0024] In some embodiments of the present application, R2 and R3 together with the carbon atoms to which they are attached form a 3- to 4-membered ring and form a spiro ring with ring Q, and the 3- to 4-membered ring is a 3- to 4-membered carbon ring.

[0025] In some embodiments of the present application, R2 and R3 together with the carbon atoms to which they are attached form a 3-membered ring and form a spiro ring with ring Q, and the 3-membered ring is a 3-membered carbon ring.

[0026] In some embodiments of the present application, R a , R b , R c and R d Each of them is independently selected from H, D, CH3, and F.

[0027] In some embodiments of the present application, R e Selected from CF3, CHF2.

[0028] In some embodiments of the present application, X is selected from S.

[0029] In some embodiments of the present application, X is selected from a single bond.

[0030] In some embodiments of the present application, n is selected from 1.

[0031] In some embodiments of the present application, n is selected from 2.

[0032] In some embodiments of this application, Preferably

[0033] In some embodiments of this application, Preferably In some embodiments of this application, Preferably In some embodiments of the present application, R0 is selected from

[0034] In some embodiments of the present application, the structural unit Selected from wherein R4 is as defined above.

[0035] In some embodiments of the present application, the structural unit Selected from wherein R4 is as defined above.

[0036] The second aspect of the present application provides a specific compound, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, the compound being selected from

[0037]

[0038]

[0039] The third aspect of the present application provides a pharmaceutical composition, which comprises the compound described in the first aspect or the second aspect of the present application, its stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0040] In some embodiments of the present application, in the pharmaceutical composition, the content of the compound, stereoisomer or pharmaceutically acceptable salt thereof is 1%-95%.

[0041] In some embodiments of the present application, in the pharmaceutical composition, the pharmaceutically acceptable carrier includes one or more of a filler, a disintegrant, a binder, a glidant, and a lubricant.

[0042] The fourth aspect of the present application provides a PTPN2 inhibitor, which comprises the compound described in the first aspect or the second aspect of the present application, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in the third aspect of the present application.

[0043] The fifth aspect of the present application provides the use of the compound described in the first aspect or the second aspect of the present application, its stereoisomer or pharmaceutically acceptable salt, or the pharmaceutical composition described in the third aspect of the present application, or the PTPN2 inhibitor described in the fourth aspect of the present application in the preparation of a drug.

[0044] In certain embodiments of the present application, the drug is used to treat a PTPN2-mediated disease.

[0045] In certain embodiments of the present application, the disease is cancer.

[0046] The sixth aspect of the present application provides a method for inhibiting PTPN2 activity, comprising contacting the compound described in the first aspect or the second aspect of the present application, its stereoisomer or pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in the third aspect of the present application, or the PTPN2 inhibitor described in the fourth aspect of the present application with the PTPN2.

[0047] The seventh aspect of the present application provides a method for treating cancer, which comprises administering to a patient a therapeutically effective amount of the compound described in the first aspect or the second aspect of the present application, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in the third aspect of the present application, or the PTPN2 inhibitor described in the fourth aspect of the present application.

[0048] Technical Effects

[0049] The compounds of the present application have a good inhibitory effect on PTPN2, thereby obtaining excellent activity in inhibiting tumor growth.

[0050] Definition and Description

[0051] Unless otherwise specified, the following terms and phrases used herein are intended to have the following meanings. A particular term or phrase should not be considered to be uncertain or unclear in the absence of a special definition, but should be understood according to its ordinary meaning. When a trade name appears in this article, it is intended to refer to its corresponding commercial product or its active ingredient.

[0052] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of humans and animals without excessive toxicity, irritation, allergic response or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0053] The term "pharmaceutically acceptable salt" refers to salts of compounds of the present application, prepared from compounds with specific substituents found in the present application and relatively non-toxic acids or bases. When the compounds of the present application contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compounds of the present application contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Certain specific compounds of the present application contain basic and acidic functional groups and can be converted into either base or acid addition salts.

[0054] The pharmaceutically acceptable salts of the present application can be synthesized by conventional chemical methods from parent compounds containing acid radicals or bases. Generally, the preparation method of such salts is: in water or an organic solvent or a mixture of the two, these compounds in free acid or base form are reacted with a stoichiometric amount of an appropriate base or acid to prepare.

[0055] The compounds of the present application may exist in specific geometric or stereoisomeric forms. The present application contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as mixtures enriched in enantiomers or diastereomers, all of which fall within the scope of the present application. Additional asymmetric carbon atoms may exist in substituents such as alkyl. All of these isomers and their mixtures are included within the scope of the present application.

[0056] Unless otherwise indicated, the term "enantiomer" or "optical isomer" refers to stereoisomers that are mirror images of one another.

[0057] Unless otherwise indicated, the term "cis-trans isomers" or "geometric isomers" arises from the inability of a ring to rotate freely about double bonds or single bonds of ring carbon atoms.

[0058] Unless otherwise indicated, the term "diastereomer" refers to stereoisomers that have two or more chiral centers and that are not mirror images of each other.

[0059] Unless otherwise indicated, "(+)" indicates dextrorotatory, "(-)" indicates levorotatory, and "(±)" indicates racemic.

[0060] Unless otherwise specified, the key is a solid wedge. and dotted wedge key Indicates the absolute configuration of a stereocenter; in the absence of a stereocenter in the structure, a solid wedge bond and dotted wedge key When they appear together, they indicate relative configuration, for example, represents trans-1,4-dimethylcyclohexane.

[0061] Unless otherwise indicated, the term "tautomer" or "tautomeric form" refers to isomers of different functional groups that are in dynamic equilibrium at room temperature and can rapidly interconvert. If tautomerism is possible (such as in solution), chemical equilibrium of tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions via proton migration, such as keto-enol isomerization and imine-enamine isomerization. Valence tautomers include interconversions via reorganization of some bonding electrons. A specific example of keto-enol tautomerization is the interconversion between pentane-2,4-dione and 4-hydroxypent-3-en-2-one.

[0062] Unless otherwise indicated, the terms "enriched in one isomer", "isomerically enriched", "enriched in one enantiomer" or "enantiomerically enriched" mean that the content of one isomer or enantiomer is less than 100%, and the content of the isomer or enantiomer is greater than or equal to 60%, or greater than or equal to 70%, or greater than or equal to 80%, or greater than or equal to 90%, or greater than or equal to 95%, or greater than or equal to 96%, or greater than or equal to 97%, or greater than or equal to 98%, or greater than or equal to 99%, or greater than or equal to 99.5%, or greater than or equal to 99.6%, or greater than or equal to 99.7%, or greater than or equal to 99.8%, or greater than or equal to 99.9%.

[0063] Unless otherwise indicated, the term "isomer excess" or "enantiomeric excess" refers to the difference between the relative percentages of two isomers or two enantiomers. For example, if the content of one isomer or enantiomer is 90% and the content of the other isomer or enantiomer is 10%, the isomer or enantiomeric excess (ee value) is 80%.

[0064] Optically active (R)- and (S)-isomers and D and L isomers can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present application is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary group is cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), a diastereomeric salt is formed with an appropriate optically active acid or base, and then the diastereoisomers are separated by conventional methods known in the art, and then the pure enantiomer is recovered. In addition, the separation of enantiomers and diastereomers is usually accomplished by using chromatography, which uses a chiral stationary phase and is optionally combined with a chemical derivatization method (e.g., carbamate is generated from an amine).

[0065] The compounds of the present application may contain unnatural proportions of atomic isotopes on one or more of the atoms constituting the compound. For example, the compounds may be labeled with radioactive isotopes, such as tritium ( 3 H), iodine-125( 125 I) or C-14( 14 C). For another example, deuterium can be used to replace hydrogen to form a deuterated drug. The bond between deuterium and carbon is stronger than the bond between ordinary hydrogen and carbon. Compared with undeuterated drugs, deuterated drugs have the advantages of reducing toxic side effects, increasing drug stability, enhancing efficacy, and extending the biological half-life of drugs. All isotopic composition changes of the compounds of the present application, whether radioactive or not, are included in the scope of the present application.

[0066] "Optional" or "optionally" means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where said event or circumstance occurs and instances where it does not.

[0067] The term "substituted" means that any one or more hydrogen atoms on a particular atom are replaced by a substituent, which may include a variant of deuterium and hydrogen, as long as the valence state of the particular atom is normal and the substituted compound is stable. When the substituent is oxygen (i.e., =O), it means that two hydrogen atoms are replaced. Oxygen substitution does not occur on aromatic groups. The term "optionally substituted" means that it may be substituted or not substituted, and unless otherwise specified, the type and number of the substituents may be arbitrary on the basis of chemical achievable.

[0068] When any variable (e.g., R) occurs more than once in a compound's composition or structure, its definition at each occurrence is independent. Thus, for example, if a group is substituted with 0-2 Rs, the group may be optionally substituted with up to two Rs, and each occurrence of R is an independent choice. In addition, combinations of substituents and / or variants thereof are permitted only if such combinations result in stable compounds.

[0069] When the number of a linking group is 0, such as -(CRR)0-, it means that the linking group is a single bond.

[0070] When one of the variables is selected from a single bond, it means that the two groups it connects are directly connected. For example, when L in ALZ represents a single bond, it means that the structure is actually AZ.

[0071] When a substituent is vacant, it means that the substituent does not exist. For example, when X in AX is vacant, it means that the structure is actually A. When the listed substituent does not specify which atom it is connected to the substituted group through, the substituent can be bonded through any atom of it. For example, pyridyl as a substituent can be connected to the substituted group through any carbon atom on the pyridine ring.

[0072] When the linking group is listed without specifying its connection direction, its connection direction is connected in the same direction as reading from left to right, for example, The connecting group L is -MW-, in which case -MW- connects ring A and ring B in the same direction as the reading order from left to right to form Combinations of linkers, substituents, and / or variations thereof are permissible only if such combinations result in stable compounds.

[0073] Unless otherwise specified, when a group has one or more connectable sites, any one or more sites of the group can be connected to other groups through chemical bonds. When the chemical bond connection mode is non-positional and there are H atoms at the connectable sites, when the chemical bonds are connected, the number of H atoms at the site will decrease with the number of connected chemical bonds to become a group with the corresponding valence. The chemical bond connecting the site to other groups can be a straight solid bond. Straight dotted key or wavy line For example, the straight solid bond in -OCH3 indicates that it is connected to other groups through the oxygen atom in the group; The straight dashed bond in the group indicates that the two ends of the nitrogen atom in the group are connected to other groups; The wavy line in the phenyl group indicates that it is connected to other groups through the carbon atoms at positions 1 and 2 in the phenyl group; The straight dashed bonds in the Q ring represent the connection to other groups through the carbon atoms at positions 1 and 2 in the Q ring.

[0074] Unless otherwise specified, the term “C 1-6 "Alkyl" is used to refer to a straight or branched chain saturated hydrocarbon group consisting of 1 to 6 carbon atoms. 1-6 Alkyl groups include C 1-5 , C 1-4 , C 1-3 , C 1-2 , C 2-6 , C 2-4 , C6 and C5 alkyl, etc.; it can be monovalent (such as methyl), divalent (such as methylene) or polyvalent (such as methine). 1-6 Examples of alkyl groups include, but are not limited to, methyl (Me), ethyl (Et), propyl (including n-propyl and isopropyl), butyl (including n-butyl, isobutyl, s-butyl and t-butyl), pentyl (including n-pentyl, isopentyl and neopentyl), hexyl, and the like.

[0075] Unless otherwise specified, the term "halogen" means a fluorine, chlorine, bromine or iodine atom.

[0076] Unless otherwise specified, the term "carbocycle" refers to saturated, partially saturated or unsaturated monocyclic and polycyclic rings, and "polycyclic rings" include spirocyclic rings, condensed rings or bridged rings. The number of atoms in the ring is usually defined as the number of ring members, for example, "4-10 membered carbocycle" refers to a ring formed by 4-10 carbon atoms arranged around it, and "3-membered carbocycle" refers to a ring formed by 3 carbon atoms arranged around it, each ring is optionally substituted by an R group, and R is a group defined in the text.

[0077] The compounds of the present application can be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthesis methods, and equivalent substitution methods well known to those skilled in the art. Preferred embodiments include but are not limited to the examples of the present application.

[0078] The structure of the compounds of the present application can be confirmed by conventional methods known to those skilled in the art. If the present application involves the absolute configuration of the compound, the absolute configuration can be confirmed by conventional technical means in the art. For example, single crystal X-ray diffraction (SXRD) is used to collect diffraction intensity data of the cultured single crystal using a Bruker D8 venture diffractometer, with the light source being CuKα radiation and the scanning mode being: After scanning and collecting relevant data, the crystal structure is further analyzed using the direct method (Shelxs97) to confirm the absolute configuration.

[0079] The solvents used in this application are commercially available.

[0080] Unless otherwise specified, the reagent ratios used in silica gel column chromatography, silica gel chromatographic column chromatography and silica gel thin layer chromatography plates in this application are all volume ratios.

[0081] The following abbreviations are used in this application: HATU stands for O-(7-azabenzotriazol-1-yl)-N,N,N',N'-tetramethyluronium hexafluorophosphate; DMSO stands for dimethyl sulfoxide; CD3OD stands for deuterated methanol; CDCl3 stands for deuterated chloroform; TBSO stands for tert-butyldimethylsilyloxy.

[0082] Compounds are named according to the conventional nomenclature in the art or using The software names were used, and commercially available compounds were named using the supplier's catalog names.

[0083] Experimental Instruments Summary

[0084] The structure of the compound of the present application is determined by nuclear magnetic resonance (NMR) or / and liquid chromatography-mass spectrometry (LC-MS), or ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS). NMR chemical shift (δ) is given in parts per million (ppm). NMR is measured using a Bruker Neo 400M or Bruker Ascend 400 nuclear magnetic instrument, and the measurement solvent is deuterated dimethyl sulfoxide (DMSO-d6), deuterated methanol (CD3OD) and deuterated chloroform (CDCl3), heavy water (D2O), and the internal standard is tetramethylsilane (TMS).

[0085] Liquid chromatography-mass spectrometry (LC-MS) was performed using an Agilent 1260-6125B single quadrupole massspectrometer with a Welch Biomate column (C18, 2.7 um, 4.6*50 mm) or a Waters H-ClassSQD2 with a Welch Ultimate column (XB-C18, 1.8 um, 2.1*50 mm) mass spectrometer (the ion source was electrospray ionization).

[0086] Ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS) was performed using a Waters UPLC H-class SQD mass spectrometer (with electrospray ionization as the ion source).

[0087] HPLC determination was performed using Waters e2695-2998 or Waters ARC and Agilent 1260 or Agilent Poroshell HPH high performance liquid chromatography.

[0088] Preparative HPLC used Waters 2555-2489 (10 μm, ODS 250 cm×5 cm) or GILSON Trilution LC, and the column was Welch XB-C18 column (5 um, 21.2*150 mm).

[0089] The chiral HPLC determination used waters acquity UPC2; the columns were Daicel chiralpak AD-H (5um, 4.6*250mm), Daicel chiralpak OD-H (5um, 4.6*250mm), Daicel chiralpak IG-3 (3um, 4.6*150mm), Chiral Technologies Europe AD-3 (3um, 3.0*150mm) and Trefoil TM Technology Trefoil TM AMY1 (2.5um, 3.0*150mm).

[0090] Supercritical fluid chromatography (SFC) used Waters SFC 80Q, and the column was Daicel Chiralcel OD / OJ / OZ (20×250 mm, 10 um) or Daicel Chiralpak IC / IG / IH / AD / AS (20×250 mm, 10 um).

[0091] The thin layer chromatography silica gel plate uses GF254 silica gel plate from Yantai Jiangyou Silica Gel Development Co., Ltd. or GF254 silica gel plate from Rushan Shangbang New Materials Co., Ltd. The specification used by TLC is 0.15mm-0.20mm, and the preparation type is 20x 20cm. Column chromatography generally uses 200-300 mesh silica gel from Chenghua as the carrier. DETAILED DESCRIPTION

[0092] The present application is described in detail below by examples, but it is not meant to limit the present application in any adverse way. The compounds of the present application can be prepared by a variety of synthesis methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by the combination of the specific embodiments and other chemical synthesis methods, and the equivalent substitution methods well known to those skilled in the art, and preferred embodiments include but are not limited to the embodiments of the present application. It will be apparent to those skilled in the art that various changes and improvements are made to the specific embodiments of the present application without departing from the spirit and scope of the present application.

[0093] Intermediate INT-A 5-(1-fluoro-3-hydroxy-7-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2,5-thiadiazolin-3-one 1,1-dioxide

[0094]

[0095] Reaction route:

[0096]

[0097] Steps:

[0098] Step A: At room temperature, 6-bromo-8-fluoro-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane] (5.0 g, 17.4 mmol), biboric acid pinacol ester (5.3 g, 20.8 mmol) and potassium acetate (4.3 g, 43.5 mmol) were dissolved in 1,4-dioxane (100 mL), and then [1,1'-bis(diphenylphosphino)ferrocene] dichloropalladium dichloromethane complex (0.74 g, 0.87 mmol) was added. After replacing nitrogen, the reaction solution was heated to 100 ° C and stirred for 4 hours.

[0099] The mixture was cooled to room temperature, filtered, and the filtrate was concentrated under reduced pressure to give 2-(8-fluoro-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (10.8 g, crude product).

[0100] MS (ESI) M / Z: 335.2 [M+H] + .

[0101] Step B: 2-(8-fluoro-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]-6-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (10.6 g, 31.7 mmol) was dissolved in a mixed solvent of tetrahydrofuran (100 mL) and water (30 mL) at room temperature, and then sodium perborate tetrahydrate (14.7 g, 95.4 mmol) was added, and the reaction solution was stirred at room temperature for 16 hours.

[0102] The reaction solution was filtered, water (50 mL) was added to the filtrate, and the mixture was extracted with ethyl acetate solution (80 mL×3). The organic phases were combined, washed with saturated sodium chloride solution (50 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 8-fluoro-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]-6-ol (6.0 g, crude product).

[0103] MS (ESI) M / Z: 225.2 [M+H] + .

[0104] Step C: Add benzyl bromide (3.8 g, 21.5 mmol) to a solution of 8-fluoro-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]-6-ol (6.0 g, 26.7 mmol) and cesium carbonate (21.9 g, 66.9 mmol) in N,N-dimethylformamide (100 mL) at room temperature and stir at room temperature for 3 hours.

[0105] Water (300 mL) was added to the reaction solution to quench the reaction, and the mixture was extracted with ethyl acetate solution (100 mL×3), washed with saturated sodium chloride solution (50 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The mixture was purified by normal phase column chromatography to give 6-(benzyloxy)-8-fluoro-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane] (4.2 g).

[0106] MS (ESI) M / Z: 315.1 [M+H] + .

[0107] Step D: Under ice bath, add n-butyllithium (7.5 mL, 18.6 mmol, 2.5 M in hexane) dropwise to a solution of 2,2,6,6-tetramethylpiperidine (2.8 g, 19.9 mmol) in tetrahydrofuran (100 mL), and stir under ice bath for 2 hours. The reaction solution was then cooled to -78°C, and a tetrahydrofuran solution (20 mL) of 6-(benzyloxy)-8-fluoro-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane] (3.9 g, 12.4 mmol) was slowly added dropwise. N,N,N',N'-tetramethylethylenediamine (22 g, 186 mmol) was then added to the reaction solution. The system was stirred at -78°C for 2 hours. Finally, a tetrahydrofuran solution (20 mL) of iodine (6.3 g, 24.8 mmol) was slowly added dropwise, keeping the internal temperature at no more than -65°C. The reaction solution was slowly raised to room temperature and stirred for 16 hours.

[0108] Saturated ammonium chloride solution (80 mL) was added to the reaction solution to quench the reaction, and the mixture was stirred for 15 minutes, extracted with ethyl acetate (150 mL×3), washed with saturated sodium chloride solution (50 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting mixture was purified by normal phase column chromatography to give 6-(benzyloxy)-8-fluoro-7-iodo-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane] (4.0 g).

[0109] MS (ESI) M / Z: 441.0 [M+H] + .

[0110] 1HNMR (400MHz, DMSO-d6): δ7.50(d,J=7.4Hz,2H),7.46-7.37(m,J=7.4Hz,2H),7.34(d,J=7.4Hz,1H),6 .76(s,1H),5.17(s,2H),3.99–3.88(m,4H),2.87(t,J=6.6Hz,2H),2.77(s,2H),1.84(t,J=6.6Hz,2H).

[0111] Step E: Under nitrogen protection, glycine tert-butyl ester (1.75 g, 13.4 mmol) was added to 6-(benzyloxy)-8-fluoro-7-iodo-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane] (3.9 g, 8.9 mmol), 2-(dicyclohexylphosphine)-3,6-dimethoxy-2'-4'-6'-tri-1-propyl-11'-biphenyl (0.95 g, 1.8mmol), methanesulfonic acid (2-dicyclohexylphosphine)-3,6-dimethoxy-2',4',6'-triisopropyl-1,1'-biphenyl)(2'-amino-1,1'-biphenyl-2-yl)palladium(II) (1.6g, 1.8mmol) and cesium carbonate (5.9g, 18mmol) in 1,4-dioxane solution (120mL), the reaction solution was heated to 60°C and stirred for 16 hours.

[0112] The reaction solution was cooled to room temperature, quenched with saturated aqueous ammonium chloride solution (50 mL), diluted with water (30 mL), filtered, and the filtrate was extracted with ethyl acetate (80 mL×3). The organic phases were combined, washed with saturated sodium chloride solution (50 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting mixture was purified by normal phase column chromatography to give (6-(benzyloxy)-8-fluoro-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]-7-yl)glycine tert-butyl ester (1.6 g).

[0113] MS (ESI) M / Z: 444.4 [M+H] + .

[0114] Step F: In an ice bath, a solution of allyl alcohol (0.31 g, 5.4 mmol) in dichloromethane (5 mL) was slowly added dropwise to a solution of chlorosulfonyl isocyanate (0.76 g, 5.4 mmol) in dichloromethane (10 mL). The reaction solution was stirred for 1 hour, cooled to 0°C, and a solution of (6-(benzyloxy)-8-fluoro-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]-7-yl)glycine tert-butyl ester (1.6 g, 3.6 mmol) and N,N-diisopropylethylamine (0.92 g, 7.2 mmol) in dichloromethane (15 mL) was slowly added dropwise. The reaction solution was heated to room temperature and stirred for 2 hours.

[0115] The mixture was quenched with water (50 mL), extracted with dichloromethane (50 mL×3), the organic phases were combined, washed with saturated sodium chloride solution (50 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting mixture was purified by normal phase column chromatography to give the target product, N-(N-((allyloxy)carbonyl)aminosulfonyl)-N-(6-(benzyloxy)-8-fluoro-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]-7-yl)glycine tert-butyl ester (1.0 g).

[0116] MS (ESI) M / Z: 629.3 [M+Na] + .

[0117] Step G: Under nitrogen protection, dissolve N-(N-((allyloxy)carbonyl)aminosulfonyl)-N-(6-(benzyloxy)-8-fluoro-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]-7-yl)glycine tert-butyl ester (1.0 g, 1.6 mmol) in methanol (15 mL), and then slowly add a solution of sodium tert-butoxide (0.48 g, 5.0 mmol) in tetrahydrofuran (6 mL) dropwise. After stirring at room temperature for 5 minutes, tetrakis(triphenylphosphine)palladium (0.18 g, 0.16 mmol) is added and the reaction solution is heated to 60°C and stirred for 2 hours.

[0118] The reaction solution was cooled to room temperature, diluted with ethyl acetate (50 mL), and concentrated under reduced pressure. The resulting mixture was dissolved with ethyl acetate (50 mL), and hydrochloric acid (10 mL, 1 M) was slowly added dropwise under ice bath conditions to quench (pH <2). The mixture was extracted with ethyl acetate (30 mL×3), and the organic phases were combined, washed with saturated sodium chloride solution (50 mL×2), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting mixture was purified by normal phase column chromatography to obtain the target product 5-(6-(benzyloxy)-8-fluoro-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]-7-yl)-1,2,5-thiadiazoline-3-1,1-dioxide (750.0 mg).

[0119] MS (ESI) M / Z: 449.1 [M+H] + .

[0120] 1 H-NMR (400MHz, DMSO-d6): δ7.49 (d, J=7.0Hz, 2H), 7.39-7.26 (m, 3H), 6.73 (s, 1H), 5.1 1(s,2H),3.96-3.92(m,6H),2.85(t,J=6.6Hz,2H),2.72(s,2H),1.83(t,J=6.6Hz,2H).

[0121] Step H: At room temperature, 5-(6-(benzyloxy)-8-fluoro-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]-7-yl)-1,2,5-thiadiazoline-3-1,1-dioxide (600.0 mg, 1.3 mmol) was dissolved in a mixed solvent (tetrahydrofuran:methanol=1:1, 10 mL), palladium carbon (300.0 mg, 10%) was added, and after replacing the hydrogen, the reaction solution was stirred for 5 hours.

[0122] The reaction solution was filtered, the filter cake was washed with methanol (10 mL × 3), and the filtrate was concentrated in vacuo to give 5-(8-fluoro-6-hydroxy-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]-7-yl)-1,2,5-thiadiazoline-3-one 1,1-dioxide (460.0 mg) without purification.

[0123] MS (ESI) M / Z: 359.1 [M+H] + .

[0124] Step I: Dissolve 5-(8-fluoro-6-hydroxy-3,4-dihydro-1H-spiro[naphthalene-2,2'-[1,3]dioxolane]-7-yl)-1,2,5-thiadiazoline-3-one 1,1-dioxide (460 mg, 1.5 mmol) in formic acid (3 mL) at room temperature and stir the reaction mixture for 1 hour.

[0125] Dichloromethane (30 mL) was added to the reaction solution for dilution, and the mixture was concentrated in vacuo. The resulting mixture was diluted and concentrated with dichloromethane (30 mL) three more times to remove most of the formic acid to obtain 5-(1-fluoro-3-hydroxy-7-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2,5-thiadiazoline-3-one 1,1-dioxide (300.0 mg, crude product).

[0126] MS (ESI) M / Z: 315.0 [M+H] + .

[0127] Example 1

[0128] 5-(7-((2-(3-(difluoromethylene)cyclobutyl)ethyl)amino)-1-fluoro-3-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2,5-thiadiazoline-3-1,1-dioxide

[0129]

[0130] Reaction route:

[0131]

[0132] Steps:

[0133] Step A: Under nitrogen protection, zinc copper reagent (3.5 g, 54.0 mmol) was added to a solution of 4-chloro-1-butene (4.5 g, 49.1 mmol) in anhydrous ether (80 mL), and then a solution of trichloroacetyl chloride (5.8 mL, 51.5 mmol) and phosphorus oxychloride (4.8 mL, 51.5 mmol) in anhydrous ether (40 mL) was slowly added dropwise over 20 minutes. The reaction solution was heated to 40 ° C and refluxed with stirring for 16 hours.

[0134] The reaction solution was cooled to room temperature and filtered. The filter cake was washed with anhydrous ether (30 mL×3). The filtrate was vortexed to 60 mL and extracted with petroleum ether (45 mL). A brown oil was precipitated. The upper clear liquid was poured out and the brown oil was extracted with petroleum ether (45 mL×2). The clear liquids were combined, washed with water (40 mL×3), and then washed with a saturated sodium chloride solution (30 mL). It was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 2,2-dichloro-3-(2-chloroethyl)cyclobutane-1-one (7.8 g, crude product).

[0135] 1 HNMR (400MHz, CDCl3) δ3.70–3.67(m,2H),3.47–3.41(m,1H),3.25–3.04(m,2H),2.51–2.37(m,1H),2.14-2.09(m,1H).

[0136] Step B: Dissolve 2,2-dichloro-3-(2-chloroethyl)cyclobutane-1-one (7.8 g, 38.8 mmol) in acetic acid (50 mL) at room temperature, heat to 90°C, then slowly add zinc powder (8.5 g, 130.3 mmol) in batches, and stir the reaction solution at 90°C for 2 hours.

[0137] After completion, the reaction solution was cooled to room temperature and filtered. The filter cake was washed with dichloromethane (40 mL×3). The organic phase was washed with water (80 mL×2) and saturated sodium bicarbonate solution (100 mL×4), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 3-(2-chloroethyl)cyclobutane-1-one (2.5 g, crude product).

[0138] 1 HNMR (400MHz, CDCl3) δ3.58(t,J=6.6Hz,2H),3.28–3.16(m,2H),2.82–2.70(m,2H),2.69–2.56(m,1H),2.11–2.06(m,2H).

[0139] Step C: Under nitrogen protection, sodium azide (3.6 g, 55.4 mmol) was added to a solution of 3-(2-chloroethyl)cyclobutane-1-one (2.5 g, 18.5 mmol) in N,N-dimethylformamide (50 mL). The reaction solution was heated to 80°C and stirred for 16 hours.

[0140] The reaction solution was cooled to room temperature, quenched by adding water (80 mL), extracted with ethyl acetate solution (60 mL×3), washed with saturated sodium chloride solution (40 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 3-(2-azidoethyl)cyclobutane-1-one (1.5 g).

[0141] 1 HNMR (400MHz, CDCl3): δ3.37(t,J=6.6Hz,2H),3.27–3.16(m,2H),2.81–2.69(m,2H),2.57–2.44(m,1H),1.92–1.87(m,2H).

[0142] Step D: At -50 °C and under nitrogen protection, a solution of potassium tert-butoxide (2.4 g, 21.6 mmol) in N,N-dimethylformamide (10 mL) was added dropwise to a solution of 3-(2-azidoethyl)cyclobutane-1-one (1.5 g, 10.79 mmol) and 2-((difluoromethyl)sulfonyl)pyridine (2.5 g, 12.9 mmol) in N,N-dimethylformamide (60 mL), and the mixture was heated to -40 °C and stirred for 1 hour.

[0143] The mixture was slowly heated to room temperature, saturated ammonium chloride solution (30 mL) and dilute hydrochloric acid (10 mL, 1 M) were added to the reaction solution to quench, and then the system was heated to room temperature, extracted with ethyl acetate (100 mL × 3), washed with water (80 mL) and saturated sodium chloride solution (50 mL × 2) in sequence, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to about 2 ml to obtain 1-(2-azidoethyl)-3-(difluoromethylene)cyclobutane (1.4 g, the crude product contains ethyl acetate) which was used directly in the next step without purification.

[0144] Step E: At 0°C and under nitrogen protection, triphenylphosphine (1.5 g, 5.7 mmol) was added to a tetrahydrofuran solution (30 mL) of 1-(2-azidoethyl)-3-(difluoromethylene)cyclobutane (1.4 g, 8.1 mmol), the reaction solution was heated to room temperature and stirred for 16 hours, then water (1.5 mL) was added to the reaction solution, and the system was heated to 90°C and stirred under reflux for 2 hours.

[0145] The reaction solution was cooled to room temperature, and then dilute hydrochloric acid (10 mL, 1 M) was slowly added dropwise in an ice bath and stirred for 15 minutes. The impurities were extracted with dichloromethane (30 mL×3), and the aqueous phase was concentrated under reduced pressure to 2 ml. The resulting residue was purified by reverse phase to obtain 2-(3-(difluoromethylene)cyclobutyl)ethane-1-amine hydrochloride (100 mg).

[0146] MS (ESI) M / Z: 148.2 [M+H] + .1HNMR (400MHz, DMSO-d6) δ8.02(br,3H),2.84–2.73(m,2H),2.72-2.66(m,2H),2.44–2.33(m,1H),2.36–2.24(m,2H),1.74(q,J=8.0Hz,2H).

[0147] Step F: Under ice bath and nitrogen protection, 2-(3-(difluoromethylene)cyclobutyl)ethane-1-amine hydrochloride (30.0 mg, 0.2 mmol) was dissolved in anhydrous ethanol (1 mL), triethylamine (0.1 mL) was added, and the reaction solution was stirred for 30 minutes, and then acetic acid (0.15 mL) was added and stirred for 10 minutes. The above system was added to an anhydrous ethanol (1.5 mL) solution of 5-(1-fluoro-3-hydroxy-7-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2,5-thiadiazolin-3-one 1,1-dioxide (65 mg, 0.2 mmol), and then sodium cyanoborohydride (25.6 mg, 0.4 mmol) was added, and the reaction solution was heated to room temperature and stirred for 1 hour.

[0148] The reaction solution was cooled to 0°C, quenched with saturated sodium bicarbonate solution (0.5 mL), and concentrated under reduced pressure. The resulting residue was purified by high performance liquid chromatography to obtain the target product 5-(7-((2-(3-(difluoromethylene)cyclobutyl)ethyl)amino)-1-fluoro-3-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2,5-thiadiazoline-3-one 1,1-dioxide (13.8 mg). MS (ESI) M / Z: 446.3 [M+H] + .

[0149] 1 HNMR (400MHz, DMSO-d6): δ9.26(s,1H),8.46(br,1H),6.46(s,1H),3.93(s,2H),3.41(s,1H),3.09(d,J=16.0Hz, 1H),3.00-2.92(m,2H),2.86-2.64(m,5H),2.46-2.30(m,4H),2.13(s,1H),1.82-1.76(m,2H),1.70-1.55(m,1H).

[0150] Example 2

[0151] 5-(7-((2-(3-(difluoromethylene)cyclobutyl)ethyl)amino)-1-fluoro-3-hydroxy-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2,5-thiadiazoline-3-1,1-dioxide

[0152]

[0153] Reaction route:

[0154]

[0155] Steps:

[0156] Step A: To a solution of methyltriphenylphosphonium bromide (1.7 g, 4.8 mmol) in tetrahydrofuran (30 mL) was slowly added potassium tert-butoxide (4.8 mL, 4.8 mmol, 1 M tetrahydrofuran solution) at 0°C, and stirred for 20 minutes. Then, 2-(3-oxocyclobutyl)acetonitrile (400 mg, 3.7 mmol) was dissolved in tetrahydrofuran (5 mL) and added to the above solution at 0°C. The mixture was warmed to room temperature and stirred for 2 hours.

[0157] After TLC dot plate display reaction is completed, the mixture is quenched with saturated aqueous ammonium chloride solution (20mL), diluted with water, and the mixture is extracted with ethyl acetate (50mL). After collecting organic phase, it is washed with saturated brine (100mL) and dried over anhydrous sodium sulfate. Filter, and purify by column chromatography to obtain 2-(3-methylenecyclobutyl)acetonitrile (220mg) after being spin-dried.

[0158] 1 H-NMR (400MHz, DMSO-d6): δ4.81–4.76(m,2H),2.85–2.76(m,2H),2.68(d,J=7.2Hz,2H),2.64–2.53(m,1H),2.47–2.39(m,2H).

[0159] Step B: To a solution of 2-(3-methylenecyclobutyl)acetonitrile (220 mg, 2.1 mmol) in tetrahydrofuran (10 mL) was slowly added lithium aluminum tetrahydride (2.1 mL, 2.1 mmol, 1 M tetrahydrofuran solution) at 0°C, and the mixture was warmed to room temperature and stirred for 2 hours.

[0160] Sodium sulfate decahydrate (1 g) was added, stirring was continued for half an hour, and the mixture was filtered. The filtrate was concentrated under reduced pressure to give crude 2-(3-methylenecyclobutyl)ethane-1-amine (200 mg).

[0161] MS (ESI) M / Z: 112.3 [M+H] + .

[0162] Step C: Dissolve the crude 2-(3-methylenecyclobutyl)ethane-1-amine (200 mg, 1.8 mmol) in tetrahydrofuran (10 mL) at room temperature, add triethylamine (545 mg, 5.4 mmol) and di-tert-butyl carbonate (589 mg, 2.7 mmol), and stir at room temperature for 3 hours.

[0163] Water (50 mL) and ethyl acetate (30 mL) were added for extraction. The organic phase was collected, washed with saturated brine (50 mL) and dried over anhydrous sodium sulfate. Filtered, the solvent was removed by vortexing, and purified by silica gel column chromatography to obtain tert-butyl (2-(3-methylenecyclobutyl)ethyl)carbamate (100 mg).

[0164] MS (ESI) M / Z: 156.2 [M-56+H] + .

[0165] 1H-NMR (500MHz, CDCl3) δ4.77–4.69 (m, 2H), 4.48 (s, 1H), 3.08 (d, J = 6.6Hz, 2H), 2.85 -2.73(m,2H),2.34-2.24(m,2H),1.67-1.63(m,2H),1.56-1.54(m,1H),1.44(s,9H).

[0166] Step D: Dissolve tert-butyl (2-(3-methylenecyclobutyl)ethyl)carbamate (100 mg, 0.5 mmol) in a mixed solution of acetone and formic acid (1:2) (3 mL) at room temperature, and stir the mixture at room temperature overnight.

[0167] After the reaction was completed, the reaction mixture was concentrated to obtain crude 2-(3-methylenecyclobutyl)ethane-1-amine formate (40 mg).

[0168] MS (ESI) M / Z: 112.4 [M+H] + .

[0169] Step E: Dissolve the crude 2-(3-methylenecyclobutyl)ethane-1-amine formate (40 mg, 0.36 mmol) in ethanol (1.0 mL), add triethylamine (55 mg, 0.54 mmol), stir for 10 minutes, add formic acid (0.1 mL) dropwise, add 5-(1-fluoro-3-hydroxy-7-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2,5-thiadiazol-3-one-1,1-carbon dioxide (136 mg, 0.43 mmol) to the mixture, stir for 20 minutes, add sodium cyanoborohydride (34 mg, 0.54 mmol) in an ice bath, warm to room temperature and stir for 1 hour.

[0170] Saturated sodium bicarbonate (0.5 mL) was added to quench the reaction, and the ethanol was removed by concentration under reduced pressure. The crude product was concentrated under reduced pressure to obtain the target product 5-(1-fluoro-3-hydroxy-7-((2-(3-methylenecyclobutyl)ethyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2,5-thiadiazolidin-3-one-1,1-carbon dioxide (3.7 mg) in the liquid phase.

[0171] MS (ESI) M / Z: 410.2 [M+H] + .

[0172] 1H NMR (400MHz, DMSO-d6): δ9.28(s,1H),8.45(s,1H),6.46(s,1H),4.76(d,J=2.4Hz,2H),3.93(s,2H),3.12-3.08(m,2H),3. 01-2.92(m,2H),2.82-2.67(m,4H),2.41-2.24(m,4H),2.15-2.13(m,2H),1.81-1.72(m,J=7.2Hz,2H),1.71-1.63(m,1H).

[0173] Example 3

[0174] 5-(1-Fluoro-3-hydroxy-7-((2-(3-(propan-2-ylidene)cyclobutyl)ethyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2,5-thiadiazolin-3-one 1,1-dioxide

[0175]

[0176] Reaction route:

[0177]

[0178] Steps:

[0179] Step A: Under nitrogen protection, zinc copper reagent (3.5 g, 54.0 mmol) was added to a solution of 4-chloro-1-butene (4.5 g, 49.1 mmol) in anhydrous ether (80 mL), and then a solution of trichloroacetyl chloride (5.8 mL, 51.5 mmol) and phosphorus oxychloride (4.8 mL, 51.5 mmol) in anhydrous ether (40 mL) was slowly added dropwise over 20 minutes. The reaction solution was heated to 40 ° C and refluxed with stirring for 16 hours.

[0180] The reaction solution was cooled to room temperature and filtered. The filter cake was washed with anhydrous ether (30 mL×3). The filtrate was vortexed to 60 mL and extracted with petroleum ether (45 mL). A brown oil was precipitated. The upper clear liquid was poured out and the brown oil was extracted with petroleum ether (45 mL×2). The clear liquids were combined, washed with water (40 mL×3), and then washed with a saturated sodium chloride solution (30 mL). It was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 2,2-dichloro-3-(2-chloroethyl)cyclobutane-1-one (7.8 g, crude product).

[0181] 1H-NMR (400MHz, CDCl3): δ3.70-3.67(m,2H),3.47-3.41(m,1H),3.25-3.04(m,2H),2.51-2.37(m,1H),2.14-2.09(m,1H).

[0182] Step B: Dissolve 2,2-dichloro-3-(2-chloroethyl)cyclobutane-1-one (7.8 g, 38.8 mmol) in acetic acid (50 mL) at room temperature, heat to 90°C, then slowly add zinc powder (8.5 g, 130.3 mmol) in batches, and stir the reaction solution at 90°C for 2 hours.

[0183] After completion, the reaction solution was cooled to room temperature and filtered. The filter cake was washed with dichloromethane (40 mL×3). The organic phase was washed with water (80 mL×2) and saturated sodium bicarbonate solution (100 mL×4), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 3-(2-chloroethyl)cyclobutane-1-one (2.5 g, crude product).

[0184] 1 H-NMR (400MHz, CDCl3): δ3.58(t,J=6.6Hz,2H),3.28–3.16(m,2H),2.82–2.70(m,2H),2.69–2.56(m,1H),2.11–2.06(m,2H).

[0185] Step C: Under nitrogen protection, sodium azide (3.6 g, 55.4 mmol) was added to a solution of 3-(2-chloroethyl)cyclobutane-1-one (2.5 g, 18.5 mmol) in N,N-dimethylformamide (50 mL). The reaction solution was heated to 80°C and stirred for 16 hours.

[0186] The reaction solution was cooled to room temperature, quenched by adding water (80 mL), extracted with ethyl acetate solution (60 mL×3), washed with saturated sodium chloride solution (40 mL×3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give 3-(2-azidoethyl)cyclobutane-1-one (1.5 g).

[0187] 1 HNMR (400MHz, CDCl3): δ3.37(t,J=6.6Hz,2H),3.27–3.16(m,2H),2.81–2.69(m,2H),2.57–2.44(m,1H),1.92–1.87(m,2H).

[0188] Step D: Under nitrogen protection at -60°C, isopropylmagnesium chloride (8.1 mL, 16.18 mmol) was added dropwise to a solution of 3-(2-azidoethyl)cyclobutane-1-one (1.5 g, 10.79 mmol) in tetrahydrofuran (20 mL). The system was stirred at -60°C for 2 hours.

[0189] The reaction solution was slowly warmed to room temperature, and then a saturated aqueous solution of ammonium chloride (20 mL) was added. The mixture was extracted with ethyl acetate (50 mL × 3), washed with water (30 mL) and a saturated sodium chloride solution (30 mL) in sequence, the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The resulting mixture was separated by thin layer chromatography to obtain 3-(2-azidoethyl)-1-isopropylcyclobutane-1-ol (330 mg).

[0190] 1 H NMR (500MHz, DMSO-d6): δ4.55 (s, 1H), 2.42-2.36 (m, 1H), 2.23–2.18 (m, 2H), 1.69–1.55 (m, 7H), 0.83 (d, J = 6.8Hz, 6H).

[0191] Step E: 3-(2-azidoethyl)-1-isopropylcyclobutane-1-ol (280 mg, 1.53 mmol) was dissolved in anhydrous toluene (10 mL), p-toluenesulfonic acid (130.7 mg, 0.76 mmol) was added, and the system was heated to 110° C. and refluxed for 16 hours.

[0192] The reaction solution was cooled to room temperature, and then water (10 mL) was added. It was extracted with ethyl acetate (10 mL × 3), washed with water (10 mL) and saturated sodium chloride solution (10 mL) in sequence, and the organic phase was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to obtain 1-(2-azidoethyl)-3-(prop-2-ylidene)cyclobutane (250 mg). This compound was used directly in the next step.

[0193] Step F: At 0°C and under nitrogen protection, triphenylphosphine (396.1 mg, 1.51 mmol) was added to a tetrahydrofuran solution (10 mL) of 1-(2-azidoethyl)-3-(propan-2-ylidene)cyclobutane (250 mg, 1.51 mmol), the reaction solution was heated to room temperature and stirred for 16 hours, then water (1.5 mL) was added to the reaction solution, and the system was heated to 90°C and stirred under reflux for 2 hours.

[0194] The reaction solution was cooled to room temperature, and then dilute hydrochloric acid (10 mL, 1 M) was slowly added dropwise in an ice bath and stirred for 15 minutes. The impurities were extracted with dichloromethane (30 mL×3), and the aqueous phase was concentrated under reduced pressure to 2 ml. The resulting residue was purified by reverse phase column chromatography to obtain 2-(3-(propan-2-ylidene)cyclobutyl)ethane-1-amine (50 mg).

[0195] 1 H NMR (400MHz, DMSO-d6): δ8.39(br,2H),2.71–2.66(m,2H),2.38–2.34(m,2H),2.29(s,6H),2.18(s,2H),1.94–1.88(m,3H).

[0196] Step G: Under ice bath and nitrogen protection, 2-(3-(propan-2-ylidene)cyclobutyl)ethane-1-amine (50.0 mg, 0.36 mmol) was dissolved in anhydrous ethanol (1 mL), triethylamine (0.1 mL) was added, and the reaction solution was stirred for 30 minutes, and then acetic acid (0.15 mL) was added and stirred for 10 minutes. Under ice bath and nitrogen protection, the above system was added to an anhydrous ethanol (1.5 mL) solution of 5-(1-fluoro-3-hydroxy-7-oxo-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2,5-thiadiazolin-3-one 1,1-dioxide (135.0 mg, 0.43 mmol), and then sodium cyanoborohydride (45.2 mg, 0.72 mmol) was added, and the reaction solution was heated to room temperature and stirred for 1 hour.

[0197] The reaction solution was cooled to 0°C, quenched with saturated sodium bicarbonate solution (1 mL), and concentrated under reduced pressure. The resulting residue was purified by HPLC to give the target product 5-(1-fluoro-3-hydroxy-7-((2-(3-(propan-2-ylidene)cyclobutyl)ethyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2,5-thiadiazoline-3-one 1,1-dioxide (12.3 mg).

[0198] MS (ESI) M / Z: 438.3 [M+H] + .

[0199] 1H NMR (400MHz, DMSO-d6): δ9.26(br,1H),8.17(br,1H),3.99-3.85(m,2H),3.09-3.05(m,1H),3.01-2.91(m,2H) ,2.78-2.72(m,2H),2.49-2.29(m,5H),2.24-2.07(m,2H),1.98-1.93(m,2H),1.68-1.65(m,3H),1.57(s,6H).

[0200] Example 4

[0201] (R)-5-(1-Fluoro-3-hydroxy-7-((2-(3-methylenecyclobutyl)ethyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl))-1,2,5-thiadiazolidin-3-one-1,1-dioxide

[0202]

[0203] Steps:

[0204] The racemic compound (3.1 g) of Example 2 was separated by chiral supercritical fluid chromatography. Separation conditions: chiral column: IC25×250 mm, 10 μm (Daicel); mobile phase: carbon dioxide / [methanol (0.5% ammonia (7M methanol solution)): acetonitrile = 1:1] = 45 / 55 (v / v); flow rate: 120 mL / min; cycle time: 4 minutes; detection wavelength: 214 nm; P1 (1.2 g, peak time 1.404 min) and P2 (1.3 g, peak time 2.169 min) were obtained. P2 was dissolved again and sent to high performance liquid phase to prepare (R)-5-(1-fluoro-3-hydroxy-7-((2-(3-methylenecyclobutyl)ethyl)amino)-5,6,7,8-tetrahydronaphthalen-2-yl)-1,2,5-thiadiazolidin-3-one-1,1-dioxide (968.3 mg).

[0205] MS (ESI) M / Z: 410.2 [M+H] + .

[0206] 1H NMR (400MHz, DMSO-d6) δ9.28 (s, 1H), 8.45 (s, 1H), 6.46 (s, 1H), 4.76 (d, J = 2.4Hz, 2H), 3.93 (s, 2H), 3.12-3.07 (m, 1H), 3. 01-2.92(m,2H),2.82-2.67(m,2H),2.41-2.24(m,3H),2.21-2.14(m,1H),1.81-1.72(m,J=7.2Hz,2H),1.71-1.63(m,1H).

[0207] Biological test evaluation:

[0208] PTPN2 in vitro enzymatic assay

[0209] This experiment detects the inhibitory effect of the test compound on the PTPN2 phosphatase activity. ABBV-CLS-484 was used as the positive control compound, and the PTPN2 compound was graded diluted and tested in duplicate. In this experiment, the DiFMUP assay method was used to detect the compound on PTPN2.

[0210] 1. Experimental Materials

[0211] 1.1 Reagents

[0212] PTPN2(TC-PTP),active(Signalchem,Cat.No.P29-20G)

[0213] DiFMUP (Invitrogen, Cat. No. D6567) (6,8-difluoro-4-methylumbelliferyl phosphate)

[0214] HEPES (pH 7.0) (HEPES buffer)

[0215] KCl (Potassium Chloride)

[0216] EDTA (ethylenediaminetetraacetic acid)

[0217] BSA (Bovine Serum Albumin)

[0218] DDT (dithiothreitol)

[0219] Tween-20 (Polysorbate-20)

[0220] ABBV-CLS-484(MCE,HY-145923)

[0221] 1.2 Instrument consumables

[0222] Centrifuge (Manufacturer: Eppendorf, Model: R5430 )

[0223] Microplate reader (manufacturer: Molecular Devices, model: )

[0224] 384-well plate (Perkin Elmer, Cat. No. 6007279)

[0225] 2. Experimental Methods

[0226] 2.1 Configuration of reaction buffer:

[0227] Prepare reaction buffer: 50 mM HEPES (pH 7.0), 50 mM KCl, 2 mM EDTA, 0.02% BSA, 2 mM DDT, 0.01% Tween-20 dissolved in deionized water;

[0228] 2.2 Compound configuration

[0229] Dilute the solution with DMSO to 100 times the final concentration in a 384-well plate and set aside for later use;

[0230] 2.3 Preparation of enzyme solution

[0231] PTPN2 enzyme was added to the reaction buffer to a concentration of 0.4 nM;

[0232] 2.4 Preparation of substrate solution

[0233] Dissolve DiFMUP in the reaction buffer to a concentration of 28.86 μM;

[0234] 2.5 Phosphatase fluorescent substrate reaction

[0235] 1) Use Echo550 to transfer the compound to the 384 reaction plate, and add DMSO to the Min control well and the Max control well respectively;

[0236] 2) Add 10 μL of enzyme solution to the compound wells and Max control wells respectively; add 10 μL of reaction buffer to the Min control wells;

[0237] 3) Centrifuge and incubate at room temperature for 60 min;

[0238] 4) Add 10 μL of substrate solution to each well to start the reaction;

[0239] 5) Incubate at room temperature for 30-60 minutes after centrifugation;

[0240] 2.6 Use The plate was read at excitation at 360 nm and emission at 460 nm.

[0241] 3 Data analysis

[0242] 1) Calculation formula

[0243] %Inhibition=(Signal.max-Signal.sample / Signal.max-Signal.min)×100%

[0244] Signal.sample is the signal value of the sample well; Signal.min: the mean signal value of the Min control well; Signal.max: the mean signal value of the Max control well.

[0245] 2) Fitting the dose-effect curve

[0246] The Log value of the compound concentration was used as the X-axis and the percentage inhibition rate was used as the Y-axis. The log (inhibitor) vs. response-Variable slope of the analysis software GraphPadPrism 8 was used to fit the dose-effect curve. Thus, the IC value of each compound on the enzyme activity was obtained. 50 value.

[0247] 4PTPN2 enzyme inhibition results

[0248] The results of the inhibition of human PTPN2 protein activity by the compounds of the present application are shown in Table 1.

[0249] Table 1 PTPN2 enzyme inhibition results

[0250] serial number <![CDATA[PTPN2 IC 50 (nM)]]> Example 1 1.9 Example 2 1.6 Example 3 15.4 Example 4 0.8 ABBV-CLS-484 1.1

[0251] Conclusion: The representative compounds of this application can effectively inhibit PTPN2 enzyme activity.

[0252] 2. Pharmacokinetics in mice

[0253] Mice were used as test animals to test the pharmacokinetic behavior of the compound of the present application in plasma at a dose of 1 mg / kg intravenous injection and 10 mg / kg oral administration.

[0254] 1. Experimental plan

[0255] 1.1 Trial Drugs:

[0256] Representative compounds of this application and reference molecule ABBV-CLS-484.

[0257] 1.2 Experimental animals

[0258] Mice: Female Balb / c.

[0259] 1.3 Administration:

[0260] Free feeding, IV (intravenous push) and fasting PO (oral), IV dose is 1 mg / kg, administration volume is 5 mL / kg; PO dose is 10 mg / kg, administration volume is 10 mL / kg.

[0261] 1.4 Experimental Equipment

[0262] The centrifuge (5810R) was purchased from Eppendorf, the pipette was purchased from Eppendorf, and the vortexer was purchased from Scientific Industries.

[0263] 1.5 Sample collection

[0264] After administration, 30 μL of blood was collected from the saphenous vein of mice at 0.0833 (IV), 0.25, 0.5, 1, 2, 4, 8 and 24 hours, placed in EDTA-K2 tubes, centrifuged at 4600 rpm at 4°C for 5 minutes, and plasma was separated and stored at -80°C.

[0265] 1.6 Sample processing

[0266] 1) Add 10 μL of blank plasma to 5 μL of plasma sample and mix well. Then add 100 μL of methanol containing internal standard diclofenac to precipitate protein, shake for 10 minutes after mixing, and centrifuge at 4°C 5800 rpm for 10 minutes.

[0267] 2) Take the supernatant solution after treatment and perform LC / MS / MS analysis on the concentration of the test compound.

[0268] 1.7 Liquid phase analysis

[0269] Liquid chromatography mass spectrometry: LCMSMS-45 (Triple Quad 6500+);

[0270] Chromatographic column: waters HSS T3 (2.1×50mm, 1.8μm);

[0271] Mobile phase: A: 1 mM ammonium acetate solution containing 0.025 vol% formic acid, B: methanol solution containing 0.025 vol% formic acid and 1 mM ammonium acetate;

[0272] Flow rate: 0.6 mL / min;

[0273] Gradient: Gradient elution 0-2.5 minutes.

[0274] 2. Experimental results and analysis

[0275] The main pharmacokinetic parameters were calculated using WinNonlin software. The pharmacokinetic parameters of intravenous injection and oral administration of drugs in mice are shown in Tables 2 and 3 below; where Dose represents dose, CL represents plasma clearance, Vss represents the volume of drug distribution, and T 1 / 2 represents the half-life of the drug, AUC represents the area under the drug-time curve, C max represents the peak concentration of the drug and F represents the bioavailability.

[0276] Table 2 Pharmacokinetic parameters of representative compounds of the present application and control molecules injected intravenously into mice

[0277]

[0278] Table 3 Pharmacokinetic parameters of representative compounds of the present application and control molecules in mice after oral administration

[0279]

[0280]

[0281] Conclusion: At the same dose, the exposure of the representative compounds of the present application in mouse plasma after oral administration is significantly higher than that of the control compound ABBV-CLS-484.

[0282] The structure of the reference compound ABBV-CLS-484 is as follows:

[0283]

Claims

1. A compound represented by formula (I), a stereoisomer thereof or a pharmaceutically acceptable salt thereof, in, Q is a 5-7 membered carbon ring; R2 and R3 are each independently hydrogen, C 1-6 Alkyl; or R2 and R3 are combined to form an alkylene group, which together with the carbon atom to which they are attached form a 3- to 4-membered ring and form a condensed ring with ring Q; or R2 and R3 are combined to form an alkylene group, which together with the carbon atom to which they are attached form a 3- to 4-membered ring and form a spiro ring with ring Q; R4 is -NHR0, R0 is selected from C1-C6 alkyl, and the C1-C6 alkyl is optionally substituted by the following groups: R a , R b , R c and R d are independently selected from H, D, C 1-6 Alkyl, halogen, C 1-6 Haloalkyl; R e Selected from H, D, C 1-6 Alkyl, C 1-6 Haloalkyl; X is selected from O, S, CH2 or a single bond; n is an integer selected from 0, 1, and 2.

2. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Q is a 6-membered carbon ring.

3. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R2 and R3 are each independently hydrogen.

4. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R2 and R3 together with the carbon atom to which they are attached form a 3- to 4-membered ring and form a condensed ring with ring Q, wherein the 3- to 4-membered ring is a 3- to 4-membered carbon ring.

5. The compound according to claim 4, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R2 and R3 together with the carbon atom to which they are attached form a 3-membered ring and form a condensed ring with ring Q, wherein the 3-membered ring is a 3-membered carbon ring.

6. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R2 and R3 together with the carbon atom to which they are attached form a 3- to 4-membered ring and form a spiro ring with ring Q, wherein the 3- to 4-membered ring is a 3- to 4-membered carbon ring.

7. The compound according to claim 6, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R2 and R3 together with the carbon atom to which they are attached form a 3-membered ring and form a spiro ring with ring Q, wherein the 3-membered ring is a 3-membered carbocyclic ring.

8. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R0 is selected from C5-C6 alkyl; preferably 9. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R0 is selected from C2-C3 alkyl, the C2-C3 alkyl is replace, R a , R b , R c and R d Each is independently selected from H, D, CH3, F; n is an integer selected from 1 or 2.

10. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R0 is selected from C2-C3 alkyl, the C2-C3 alkyl is replace, R a , R b , R c and R d Each is independently selected from H, D, CH3, F; n is an integer selected from 1 or 2.

11. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R0 is selected from C2-C3 alkyl, the C2-C3 alkyl is replace; R e Selected from CF3, CHF2; X is selected from S or a single bond.

12. The compound according to claim 9, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Selected from 13. The compound according to claim 10, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Selected from 14. The compound according to claim 11, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Selected from 15. The compound according to claim 1, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: R0 is selected from 16. The compound according to any one of claims 1 to 15, its stereoisomer or a pharmaceutically acceptable salt thereof, wherein: Structural unit Selected from wherein R4 is as defined in any one of claims 1-15.

17. A compound represented by the following formula, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, which is selected from:

18. A compound according to any one of claims 1 to 17, a stereoisomer thereof or a pharmaceutically acceptable salt thereof for use as a medicament.

19. A pharmaceutical composition comprising the compound according to any one of claims 1 to 18, its stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. 20 . A PTPN2 inhibitor comprising the compound according to any one of claims 1 to 18 , its stereoisomer or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 19 .

21. Use of the compound according to any one of claims 1 to 18, its stereoisomer or pharmaceutically acceptable salt thereof, or the pharmaceutical composition according to claim 19 in the preparation of a medicament for treating cancer.

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