CN120004876A - Protein tyrosine phosphatase inhibitors and their applications - Google Patents

Abstract

The present invention discloses a protein tyrosine phosphatase inhibitor and its application. Specifically, the present invention discloses a compound represented by formula I or a pharmaceutically acceptable salt thereof, which can be used for treating cancer and the like.

Description

Protein tyrosine phosphatase inhibitors and uses thereof

Technical Field

The invention belongs to the field of medicines, and particularly relates to a protein tyrosine phosphatase inhibitor and application thereof.

Background

Cancer immunotherapy regimens targeting immune evasion mechanisms, including checkpoint blockade (e.g., PD-1/PD-L1 and CTLA-4 blocking antibodies), have been demonstrated to be effective in treating a variety of cancers, significantly improving outcome in some refractory populations to conventional therapies. However, the development of incomplete clinical responses, inherent or acquired resistance, will continue to limit the patient population that may benefit from checkpoint blockade.

Type 2 non-receptor protein tyrosine phosphatases (PTPN 2), also known as T-cell protein tyrosine phosphatases (TC-PTP), are intracellular members of the class 1 subfamily of phosphotyrosine specific phosphatases that control a number of cell-regulated processes by removing phosphate groups from tyrosine substrates. PTPN2 is widely expressed, but expressed highest among hematopoietic cells and placental cells. In humans, PTPN2 expression is posttranscriptionally controlled by the presence of two splice variants, a 45kDa form containing a nuclear localization signal at the C-terminus upstream of the splice junction and a 48kDa classical form having a C-terminal ER retention motif. The 45kDa isoform can passively infiltrate cytosol under certain cell pressure conditions. Both isoforms have an N-terminal phosphotyrosine phosphatase catalytic domain. PTPN2 negatively regulates signaling of non-receptor tyrosine kinases (e.g., JAK1, JAK 3), receptor tyrosine kinases (e.g., INSR, EGFR, CSF A R, PDGFR), transcription factors (e.g., STAT1, STAT3, STAT 5A/b), and Src family kinases (e.g., fyn, lck). As an important negative regulator of the JAK-STAT pathway, PTPN2 function is directly regulating signaling through cytokine receptors, including ifnγ. The PTPN2 catalytic domain has 74% sequence homology to PTPN1 (also called PTP 1B) and has similar enzymatic kinetics.

Data from in vivo gene screening for loss of function using CRISPR/Cas9 genome editing in a mouse B16F10 transplantable tumor model showed that deletion of the PTPN2 gene in tumor cells improved response to an immunotherapeutic regimen of GM-CSF secretion vaccine (GVAX) plus PD-1 checkpoint blockade. Loss of PTPN2 sensitizes tumors to immunotherapy by enhancing ifnγ -mediated effects on antigen presentation and growth inhibition. The same screen also shows that genes known to be involved in immune evasion (including PD-L1 and CD 47) are also depleted in immunotherapeutic selective pressure, while genes involved in ifnγ signaling pathways (including IFNGR, JAK1 and STAT 1) are enriched. These observations are directed to the putative role of therapeutic strategies that enhance ifnγ sensing and signaling in enhancing the efficacy of cancer immunotherapy regimens.

Disclosure of Invention

In one aspect of the present invention, the present invention provides a compound of formula I or a pharmaceutically acceptable salt thereof,

Wherein ring A is selected from 8 membered heterocyclyl or cyclooctyl;

r ¹ is selected from-C _1-6 alkyl, C _3-6 cycloalkyl, 5-6 membered heterocyclyl, -C _1-6 haloalkyl, -C _1-6 hydroxyalkyl, -C _1-6 aminoalkyl, -C _1-6 alkyl-C _3-6 cycloalkyl, or-C _1-6 alkyl-5-6 membered heterocyclyl;

n is selected from 0 or 1.

In some embodiments of the invention, the compound of formula I is a compound of formula II,

R ^1a is selected from H or R ¹, and the remaining variables are defined in the invention.

In some embodiments of the invention, R ¹ is selected from-C _1-3 alkyl, C _3-6 cycloalkyl or-C _1-3 alkyl-C _3-6 cycloalkyl, the remaining variables being as defined herein.

In some embodiments of the invention, R ¹ is selected from-C _1-3 alkyl, cyclopropyl or-C _1-3 alkyl-cyclopropyl, the remaining variables being as defined herein.

In some embodiments of the invention, the structure of the compound is as follows:

in some embodiments of the invention, the structure of the compound is as follows:

in some embodiments of the invention, the structure of the compound is as follows:

In another aspect of the invention, the invention provides a pharmaceutical composition comprising a compound as described above, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

In another aspect of the invention, the invention also provides the use of a compound as described above or a pharmaceutically acceptable salt thereof or a pharmaceutical composition as described above for the manufacture of a medicament for the treatment of a PTPN2 mediated disease.

In some aspects of the invention, the PTPN2 mediated disease is selected from cancer.

The invention has one of the following advantages:

1) The compound of the invention has novel structure;

2) The compounds of the invention have a strong inhibition level of PTPN 2;

3) The compounds of the present invention have a high level of oral in vivo exposure.

As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a compound of the invention that is pharmaceutically acceptable and has the pharmacological activity of the parent compound. Such salts include salts with inorganic acids such as nitric acid, phosphoric acid, carbonic acid, etc., or with organic acids such as propionic acid, caproic acid, cyclopentapropionic acid, glycolic acid, pyruvic acid, gluconic acid, stearic acid, muconic acid, etc., or salts formed when acidic protons present on the parent compound are replaced with metal ions, e.g., alkali metal ions or alkaline earth metal ions, or complex compounds with organic bases such as ethanolamine, diethanolamine, triethanolamine, N-methylglucamine, etc. Pharmaceutically acceptable salts of the invention can be synthesized from the parent compound containing an acid or base by conventional chemical methods. Typically, such salts are prepared by reacting these compounds in free acid or base form with a stoichiometric amount of the appropriate base or acid in water or an organic solvent or a mixture of both. Generally, nonaqueous media such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile are preferred. In addition to salt forms, the compounds provided herein exist in prodrug forms. Prodrugs of the compounds described herein readily undergo chemical changes under physiological conditions to convert to the compounds of the invention. In addition, prodrugs can be converted to the compounds of the present invention by chemical or biochemical methods in an in vivo environment.

The term "C _1-6 alkyl" is used to denote a straight or branched saturated hydrocarbon group consisting of 1 to 6 carbon atoms, unless otherwise specified. The C _1-6 alkyl group includes C _1-5、C_1-4、C_1-3、C_1-2、C_2-6、C_2-4、C₆ and C ₅ alkyl groups, etc., which may be monovalent (e.g., methyl), divalent (e.g., methylene), or polyvalent (e.g., methine). Examples of C _1-6 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.

The term "haloalkyl" refers to an alkyl group having up to the full valence of the halogen atom substituent (which may be the same or different), unless otherwise specified. Non-limiting examples of haloalkyl groups include -CF₃、-C₂F₅、-CHF₂、-CCl₃、-CHCl₂、-C₂Cl₅ and the like. "C _1-6 haloalkyl" refers to an alkyl group having up to 1,2,3, 4, 5, or 6 carbon atoms in a straight, branched, or cyclic form, containing up to a complete halogen atom substituent.

The term "C _1-6 hydroxyalkyl" refers to-C _1-6 alkyl-hydroxy (OH) unless otherwise specified. Preferably C _1-3 hydroxyalkyl (i.e. -C _1-3 alkyl-hydroxy) or C _1-4 hydroxyalkyl (i.e. -C _1-4 alkyl-hydroxy). Wherein the alkyl moiety is as defined above.

The term "C _1-6 aminoalkyl" refers to-C _1-6 alkyl-amine (NH ₂), unless otherwise specified. Preferably C _1-3 aminoalkyl (i.e., -C _1-3 alkyl-amino) or C _1-4 aminoalkyl (i.e., -C _1-4 alkyl-amino). Wherein the alkyl moiety is as defined above.

Unless otherwise specified, the number of atoms on a ring is generally defined as the number of ring elements, e.g., "3-6 membered ring" refers to a "ring" of 3-6 atoms arranged around a ring.

Unless otherwise specified, the term "5-6 membered heterocyclyl" by itself or in combination with other terms, denotes a saturated cyclic group consisting of 5 to 6 ring atoms, 1,2, 3 or 4 of which are heteroatoms independently selected from O, S and N, the remainder being carbon atoms, wherein the nitrogen atoms are optionally quaternized and the nitrogen and sulfur heteroatoms may optionally be oxidized (i.e. NO and S (O) _p, p is 1 or 2). It includes monocyclic and bicyclic ring systems, wherein the bicyclic ring system includes spiro, fused and bridged rings. In addition, with respect to the "5-6 membered heterocyclic group", the heteroatom may occupy the position of attachment of the heterocycloalkyl group to the remainder of the molecule. The 5-6 membered heterocyclic group includes 5-and 6-membered heterocycloalkyl groups. Examples of 5-6 membered heterocyclyl groups include, but are not limited to, pyrrolidinyl, pyrazolidinyl, imidazolidinyl, tetrahydrothiophenyl (including tetrahydrothiophen-2-yl and tetrahydrothiophen-3-yl, etc.), tetrahydrofuranyl (including tetrahydrofuran-2-yl, etc.), tetrahydropyranyl, piperidinyl (including 1-piperidinyl, 2-piperidinyl, and 3-piperidinyl, etc.), piperazinyl (including 1-piperazinyl and 2-piperazinyl, etc.), morpholinyl (including 3-morpholinyl and 4-morpholinyl, etc.), dioxanyl, dithianyl, isoxazolidinyl, isothiazolidinyl, 1, 2-oxazinyl, 1, 2-thiazinyl, hexahydropyridazinyl, homopiperazinyl, homopiperidinyl, etc.

Unless otherwise specified, "C _3-6 cycloalkyl" means a saturated cyclic hydrocarbon group consisting of 3 to 6 carbon atoms, which is a monocyclic and bicyclic ring system, and the C _3-6 cycloalkyl includes C _3-5、C_4-5 and C _5-6 cycloalkyl groups, etc., which may be monovalent, divalent or multivalent. Examples of C _3-6 cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and the like.

The compounds of formula I of the present invention may be prepared using synthetic methods known in the art or using methods known in the art in combination with the methods described herein. The solvents, temperatures, and other reaction conditions set forth herein are exemplary and may vary according to methods well known in the art. The compounds of the examples described in the present invention can be synthesized by the methods described in the examples using appropriate starting materials according to their specific structures, or by the methods similar to those described in the examples. The starting materials for the synthesis of the compounds of the examples of the present invention may be prepared by known synthetic methods or similar methods described in the literature or obtained from commercial sources. The compounds may be further resolved as desired by methods well known in the art, such as crystallization, chromatography, etc., to give stereoisomers thereof, the resolution conditions of which are readily available to those skilled in the art by conventional means or limited experimentation. By way of further illustration, the compounds of formula I of the present invention may be synthesized using methods in which the solvents, temperatures and other reaction conditions in each step may be the same or similar to those described in the examples below, or using reaction conditions known in the art.

Detailed Description

The compounds of the present invention may be prepared by a variety of synthetic methods well known to those skilled in the art, including the specific embodiments set forth below, embodiments formed by combining them with other chemical synthetic methods, and equivalents thereof known to those skilled in the art. Preferred embodiments include, but are not limited to, embodiments of the present invention.

The present invention is described in detail below by way of examples, but is not meant to be limiting in any way. The present invention has been described in detail herein, and specific embodiments thereof are also disclosed, it will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the invention without departing from the spirit and scope of the invention. The specific conditions are not noted in the examples and are carried out according to conventional conditions or conditions recommended by the manufacturer. The reagents or apparatus used were conventional products commercially available without the manufacturer's attention.

Example 1

Step 1 to a solution of imidazole (116.6 g,1.71 mol) in dichloromethane (1.17L) was slowly added dropwise a solution of thionyl chloride (37.3 mL,514.0 mmol) in DCM (370 mL) under ice-bath. The reaction mixture was stirred at room temperature for 1 hour, and then the reaction solution was cooled to-10 ℃. To the resulting suspension was added dropwise a solution of tert-butyl N- (3-hydroxypropyl) carbamate (50.0 g,285.7 mmol) in methylene chloride (500 mL) at-10 ℃. After the completion of the dropwise addition, the reaction mixture was stirred at room temperature for 2 hours. TLC showed detection of the desired spot value. After the completion of the reaction, the reaction mixture was concentrated, and the residue was purified by silica gel column chromatography to give Compound 1-2 (45.0 g, yield) 71.3％).¹H NMR(400MHz,CDCl₃)δ4.92-4.73(m,1H),3.96-3.71(m,3H),2.29-2.12(m,1H),1.82-1.66(m,1H),1.50(s,9H).

Step 2 to a mixture of Compound 1-2 (45.0 g,203.6 mmol) in acetonitrile (500 mL)/water (250 mL) was added ruthenium chloride trihydrate (53.0 mg,0.20 mmol) and sodium periodate (47.8 g,223.4 mmol) under ice-bath. The reaction mixture was stirred at room temperature for 2 hours. The reaction was quenched with water (500 mL) and extracted with ethyl acetate. The organic phases were combined, washed with water, dried and concentrated, and the crude product was purified by silica gel column chromatography to give compounds 1 to 3 (40.0 g, yield) 82.9％).¹H NMR(400MHz,CDCl₃)δ4.67(t,J＝6.0Hz,2H),4.08-3.93(m,2H),2.19-1.97(m,2H),1.54(s,9H).

Step 3 1- (benzyloxy) -5-bromo-3-fluoro-2-nitrobenzene (20.0 g,61.3 mmol) was dissolved in tetrahydrofuran (400 mL) under nitrogen, the reaction solution was cooled to-70℃and then lithium diisopropylamide (46.0 mL, 2.0M) was slowly added dropwise. The reaction solution was stirred at-70 ℃ for half an hour. Then, to the reaction solution was slowly added tetrahydrofuran (100 mL) of Compound 1-3 (18.9 g,79.7 mmol) dropwise at-70℃and the reaction solution was stirred at-70℃for 3 hours. After the reaction was completed, the reaction mixture was poured into a dilute hydrochloric acid (200 mL, 1.0M) solution in an ice-water bath to quench the reaction. The mixed solution was extracted with ethyl acetate, the organic phases were combined, washed with water, dried and concentrated, and the crude product was purified by silica gel column chromatography to give compounds 1 to 4 (21.0 g, yield) 70.9％).¹H NMR(400MHz,CDCl₃)δ7.51-7.34(m,5H),7.12(d,J＝2.0Hz,1H),5.16(s,2H),4.61(brs,1H),3.29-3.12(m,2H),2.89-2.71(m,2H),1.83-1.69(m,2H),1.45(s,9H).

Step 4 intermediate 1-4 (11.0 g,22.7 mmol) was dissolved in dioxane/water (120 mL,5/1 v/v) and E-2-ethoxyvinyl-1-boronic acid pinacol ester (13.5 g,68.2 mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (1.66 g,2.27 mmol) and cesium carbonate (22.2 g,68.2 mmol) were added sequentially. The reaction mixture was stirred under nitrogen for 3 hours at 90 ℃. The reaction mixture was cooled to room temperature and quenched with ice water. The mixed solution was extracted with ethyl acetate, the organic phases were combined, washed with water, dried and concentrated, and the crude product was purified by silica gel column chromatography to give compounds 1 to 5 (10.0 g, yield) 92.6％).¹H NMR(400MHz,CDCl₃)δ7.45-7.31(m,5H),6.79(d,J＝12.4Hz,1H),6.72(d,J＝1.2Hz,1H),5.89(d,J＝12.4Hz,1H),5.18(s,2H),4.59(brs,1H),3.95(q,J＝7.2Hz,2H),3.15(d,J＝5.6Hz,2H),2.64(t,J＝6.8Hz,2H),1.75-1.64(m,2H),1.45(s,9H),1.38(t,J＝7.2Hz,3H).

Step 5 intermediate 1-5 (10.0 g,21.1 mmol) was dissolved in dioxane (100 mL) and dioxane hydrochloride solution (33.0 mL, 4M) was added. The reaction solution was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was neutralized with an aqueous sodium hydrogencarbonate solution. The mixture was extracted with ethyl acetate (200 mL. Times.3), and the organic phase was washed successively with water and saturated brine, dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate under reduced pressure to obtain crude product. The crude product was purified by column chromatography on silica gel to give intermediate 1-6 (5.5 g, yield 60.9%). LCMS (ESI) [ M+Na ] ⁺:451.2.

Step 6 intermediate 1-6 (5.5 g,12.8 mmol) was dissolved in methanol (60 mL) at room temperature and aqueous ammonium chloride (6.9, 128.3 mmol) solution (30 mL) and zinc powder (4.2 g,64.1 mmol) were added. The reaction solution was stirred at room temperature for 2 hours. After the reaction was completed, the reaction solution was filtered, and most of methanol was removed from the filtrate by concentration under reduced pressure, followed by dilution with ethyl acetate, washing of the organic phase with water, drying over anhydrous sodium sulfate, and filtration. The filtrate was concentrated under reduced pressure to give crude intermediate 1-7 (4.4 g, yield 86.0%). LCMS (ESI) [ M+H ] ⁺: 399.2.

Step 7 intermediate 1-7 (4.2 g,10.5 mmol) was dissolved in dichloromethane (45 mL) at room temperature and pyridine (2.6 mL,31.6 mmol) and trifluoroacetic anhydride (2.9 g,13.7 mmol) were added. The reaction solution was stirred at room temperature for half an hour. After completion of the reaction, the mixture was poured into ice water, followed by extraction with methylene chloride (50 mL. Times.3), and the organic phase was washed with water, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography to give intermediate 1-8 (4.8 g, yield 92.1%). LCMS (ESI) [ M+Na ] ⁺:517.2.

Step 8 intermediate 1-8 (4.7 g,9.5 mmol) was dissolved in N, N-dimethylformamide (50 mL) under nitrogen, followed by the addition of potassium carbonate (3.94 g,28.5 mmol) and ethyl bromoacetate (2.18 g,14.2 mmol). The reaction mixture was stirred at 60 ℃ for 1 hour. The mixture was cooled to room temperature, the reaction solution was poured into ice water, and then extracted with ethyl acetate (50 ml×3), and the organic phase was washed with water, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography to give intermediate 1-9 (5.0 g, yield 92.9%). LCMS (ESI) [ M+Na ] ⁺: 589.2.

Step 9 intermediate 1-9 (4.95 g,8.73 mmol) was dissolved in methanol (55 mL) at room temperature and sodium methoxide (9.0 mL, 5.4M) in methanol was added. The reaction solution was stirred at room temperature for 1 hour. After the reaction was completed, the reaction solution was poured into ice water, followed by extraction with methylene chloride (100 ml×3), and the organic phase was washed with water, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give crude intermediate 1-10 (4.0 g, yield 97.3%). LCMS (ESI) [ M+Na ] ⁺:471.2.

Step 10 intermediate 1-10 (3.90 g,8.29 mmol) was dissolved in dichloromethane (50 mL) and N, N-diisopropylethylamine (4.1 mL,24.87 mmol) was added dropwise with stirring

(3.10 G,12.43 mmol). The mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched with ice water. The aqueous phase was extracted with dichloromethane (100 mL. Times.3). The organic phase was washed with saturated brine (20 mL). Dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography to give intermediate 1-11 (5.0 g, yield 88.2%). LCMS (ESI) [ M+Na ] ⁺: 706.2.

Step 11 intermediate 1-11 (2.0 g,2.92 mmol) was dissolved in methanol (20 mL) at room temperature and wet palladium on carbon (400 mg,3.76 mmol) was added. The reaction was stirred at room temperature for 2 hours under an atmosphere of hydrogen (15 psi). The reaction solution is filtered, and the filtrate is decompressed and concentrated to obtain crude products. The crude product was purified by column chromatography on silica gel to give intermediate 1-12 (1.20 g, yield 89.7%). LCMS (ESI) [ M+Na ] ⁺: 482.0.

Step 12 intermediate 1-12 (1.20 g,2.61 mmol) was dissolved in methanol (30 mL) at room temperature and sodium methoxide (5.0 mL, 5.4M) in methanol was added. The reaction solution was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was poured into ice water and an ammonium chloride solution, followed by extraction with methylene chloride (100 ml×3), and the organic phase was washed with water, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give crude intermediate 1-13 (1.1 g, crude). LCMS (ESI) [ M+Na ] ⁺:450.2.

Step 13 intermediate 1-13 (1.17 g,2.73 mmol) was dissolved in methanol (20 mL) at room temperature and wet palladium on carbon (500 mg) was added. The reaction was stirred under an atmosphere of hydrogen (15 psi) at 50 ℃ for 5 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to give crude intermediate 1-14 (1.20 g, crude). LCMS (ESI) [ M+Na ] ⁺:452.2.

Step 14. Intermediate 1-14 (200 mg,0.47 mmol) was dissolved in dichloromethane (10 mL) and trifluoroacetic acid (3.0 mL) was added. The reaction solution was stirred at room temperature for 1 hour, and the solvent was dried by spin-drying to obtain a crude product. The crude product was purified by reverse phase high performance liquid chromatography to give 5- (7-fluoro-9-hydroxy-1, 2,3,4,5, 6-hexahydrobenzo [ d ] azocin-8-yl) -1,2, 5-thiadiazolidin-3-one 1, 1-dioxide (compound 1) (36.7 mg, yield 23.9％).LCMS(ESI)[M+H]⁺:330.1.¹H NMR(400MHz,DMSO-d₆)δ9.44-9.20(m,1H),8.46(brs,2H),6.63(s,1H),3.95(s,2H),3.18-3.10(m,2H),2.98-2.87(m,4H),2.81-2.72(m,2H),1.88-1.76(m,2H).

Example 2

Step 1 Compound 1 (90 mg,0.27 mmol) was dissolved in methanol (10 mL) under nitrogen, and paraformaldehyde (82.1 mg,2.73 mmol) and two drops of glacial acetic acid were added. The reaction mixture was stirred at 40 ℃ for 1 hour. Sodium cyanoborohydride (85.8 mg,1.37 mmol) was added in portions and the reaction stirred at 40℃for a further 12 hours. The mixture was cooled to room temperature, quenched by addition of ice water (10 mL), and the reaction mixture was concentrated under reduced pressure to give the crude product. The crude product is prepared, separated and purified by reverse phase high performance liquid chromatography to obtain 5- (7-fluoro-9-hydroxy-3-methyl-1, 2,3,4,5, 6-hexahydrobenzo [ d ] azocine-8-yl) -1,2, 5-thiadiazolidin-3-one 1, 1-dioxide (compound 2) (12.3 mg, yield 13.1％).LCMS(ESI)[M+H]⁺:344.1.¹H NMR(400MHz,DMSO-d₆)δ9.77(brs,1H),8.89(brs,1H),6.66(s,1H),4.14(s,2H),3.26-3.01(m,5H),2.96-2.88(m,1H),2.85-2.72(m,5H),1.99-1.74(m,2H).

Example 3

The compound of this example can be prepared using a synthetic method similar to that described in example 2. LCMS (ESI)

[M+H]⁺:358.2.¹H NMR(400MHz,DMSO-d₆)δ9.54(s,1H),8.66(s,1H),6.66(s,1H),4.04(s,2H),3.26-2.86(m,8H),2.83-2.75(m,2H),1.95-1.79(m,2H),1.20(t,J＝7.2Hz,3H).

Example 4

The compound of this example can be prepared using a synthetic method similar to that described in example 2. LCMS (ESI)

[M+H]⁺:370.2.¹H NMR(400MHz,D₂O)δ6.64(s,1H),4.28(s,2H),3.23-3.17(m,1H),3.14-3.08(m,1H),3.02-2.94(m,2H),2.92-2.86(m,1H),2.77-2.72(m,1H),2.51-2.42(m,1H),1.82(s,2H),1.24-1.14(m,3H),0.68-0.59(m,3H).

Example 5

The compound of this example can be prepared using a synthetic method similar to that described in example 2. LCMS (ESI)

[M+H]⁺:384.2.¹H NMR(400MHz,DMSO-d₆)δ6.48(s,1H),6.08(s,1H),3.96(s,2H),2.78-2.73(m,2H),2.65-2.60(m,4H),2.38-2.31(m,4H),1.52(s,2H),0.76(s,1H),0.45-0.36(m,2H),0.07-0.00(m,2H).

Example 6

Step 1- (benzyloxy) -5-bromo-3-fluoro-2-nitrobenzene (30.0 g,92.0 mmol) was dissolved in tetrahydrofuran (300 mL) under nitrogen, the reaction solution was cooled to-70℃and then lithium diisopropylamide (92.0 mL, 2.0M) was slowly added dropwise. The reaction solution was stirred at-70 ℃ for half an hour. 3-Boc-1,2, 3-oxathiazolidine 2, 2-dioxide (24.6 g,110.3 mmol) in tetrahydrofuran (100 mL) was then slowly added dropwise to the reaction at-70℃and the reaction stirred at-70℃for 3 hours. After the reaction was completed, the reaction solution was poured into a saturated ammonium chloride solution in an ice-water bath to quench the reaction. The mixed solution was extracted with ethyl acetate, and the organic phases were combined, washed with water, dried and concentrated, and the obtained crude product was purified by silica gel column chromatography to give compound 6-2 (35.0 g, yield 81.1%). LCMS (ESI) [ M+Na ] ⁺: 491.0.

Step 2 intermediate 6-2 (25.0 g,53.2 mmol) was dissolved in dioxane/water (120 mL,5/1 v/v) and E-2-ethoxyvinyl-1-boronic acid pinacol ester (52.8 g,266.3 mmol), [1,1' -bis (diphenylphosphino) ferrocene ] palladium dichloride (1.9 g,2.66 mmol) and cesium carbonate (52.1 g,159.8 mmol) were added sequentially. The reaction mixture was stirred under nitrogen for 3 hours at 90 ℃. The reaction mixture was cooled to room temperature and quenched with ice water. The mixed solution was extracted with ethyl acetate, and the organic phases were combined, washed with water, dried and concentrated, and the obtained crude product was purified by silica gel column chromatography to give compound 6-3 (17.5 g, yield 71.3%). LCMS (ESI) [ M+Na ] ⁺:483.4.

Step 3 intermediate 6-3 (17.0 g,36.9 mmol) was dissolved in dioxane (150 mL) and dioxane hydrochloride solution (50 mL, 4M) was added. The reaction solution was stirred at room temperature for 1 hour. After completion of the reaction, the reaction solution was neutralized with an aqueous sodium hydrogencarbonate solution. The mixture was extracted with ethyl acetate (300 mL. Times.3), and the organic phase was washed successively with water and saturated brine, dried over anhydrous sodium sulfate, and filtered. Concentrating the filtrate under reduced pressure to obtain crude product. The crude product was purified by column chromatography on silica gel to give intermediate 6-4 (8.5 g, yield 55.6%). LCMS (ESI) [ M+Na ] ⁺: 437.4.

Step 4 intermediate 6-4 (17.1 g,41.2 mmol) was dissolved in a mixed solvent of tetrahydrofuran (100 mL) and methanol (100 mL) at room temperature, and saturated ammonium chloride solution (50 mL) and zinc powder (13.5 g,206.3 mmol) were added. The reaction solution was stirred at room temperature for 3 hours. After the reaction was completed, the reaction solution was filtered, and most of methanol was removed from the filtrate by concentration under reduced pressure, followed by dilution with methylene chloride, washing of the organic phase with water, drying over anhydrous sodium sulfate, and filtration. The filtrate was concentrated under reduced pressure to give crude intermediate 6-5 (12.5 g, yield 78.8%). LCMS (ESI) [ M+H ] ⁺:385.0.

Step 5 intermediate 6-5 (12.5 g,32.5 mmol) was dissolved in dichloromethane (150 mL) at room temperature and pyridine (6.56 mL,81.2 mmol) and trifluoroacetic anhydride (13.7 g,65.0 mmol) were added. The reaction solution was stirred at room temperature for 2 hours. After completion of the reaction, the mixture was poured into ice water, followed by extraction with ethyl acetate (300 mL. Times.3), and the organic phase was washed with water, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography to give intermediate 6-6 (15.0 g, yield 96.0%). LCMS (ESI) [ M+Na ] ⁺: 503.0.

Step 6 intermediate 6-6 (13.0 g,27.0 mmol) was dissolved in N, N-dimethylformamide (130 mL) under nitrogen, followed by the addition of potassium carbonate (11.2 g,81.2 mmol) and methyl bromoacetate (5.0 g,32.4 mmol). The reaction mixture was stirred at 60 ℃ for 14 hours. The mixture was cooled to room temperature, the reaction solution was poured into ice water, and then extracted with ethyl acetate (300 ml×3), and the organic phase was washed with water, dried over anhydrous sodium sulfate, and filtered. The filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography to give intermediate 6-7 (13.8 g, yield 92.3%). LCMS (ESI) [ M+Na ] ⁺.0.

Step 7 intermediate 6-7 (13.8 g,25.0 mmol) was dissolved in methanol (120 mL) at room temperature and sodium methoxide (9.0 mL, 5.4M) in methanol was added. The reaction solution was stirred at room temperature for 1 hour. After completion of the reaction, the reaction mixture was poured into ice water, neutralized with saturated ammonium chloride, then extracted with methylene chloride (100 ml×3), and the organic phase was washed with water, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give crude intermediate 6-8 (10.4 g, yield 91.6%). LCMS (ESI) [ M+H ] ⁺:457.2.

Step 8 intermediate 6-8 (5.0 g,10.9 mmol) was dissolved in dichloromethane (50 mL) at room temperature, N-diisopropylethylamine (5.4 mL,32.8 mmol) was added and N- (benzyloxycarbonyl) sulfonyl chloride (4.1 g,16.4 mmol) was slowly added dropwise with stirring. The mixture was stirred at room temperature for 1 hour. The reaction mixture was quenched with ice water. The aqueous phase was extracted with dichloromethane (100 mL. Times.3). The organic phase was washed with saturated brine. Dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography to give intermediate 6-9 (6.1 g, yield 83.2%). LCMS (ESI) [ M-Boc+H ] ⁺: 570.2.

Step 9 intermediate 6-9 (8.1 g,12.1 mmol) was dissolved in isopropanol (100 mL) at room temperature and wet palladium on carbon (810 mg,7.61 mmol) was added. The reaction was stirred under an atmosphere of hydrogen (15 psi) at 40 ℃ for 3 hours. The reaction solution was filtered, and the filtrate was concentrated under reduced pressure to give crude intermediate 6-10 (4.7 g, yield 86.8%). LCMS (ESI)

[M+Na]⁺:470.4。

Step 10 intermediate 6-10 (4.1 g,9.16 mmol) was dissolved in methanol (40 mL) at room temperature and sodium methoxide (10.0 mL, 5.4M) in methanol was added. The reaction was stirred at 40 ℃ for 1 hour. After completion of the reaction, the reaction solution was poured into ice water and an ammonium chloride solution, followed by extraction with ethyl acetate (200 ml×3), and the organic phase was washed with water, dried over anhydrous sodium sulfate and filtered. The filtrate was concentrated under reduced pressure to give a crude product, which was purified by silica gel column chromatography to give intermediate 6-11 (3.2 g, yield 84.1%). LCMS (ESI) [ M+Na ] ⁺:438.0.

Step 11 intermediate 6-11 (5.4 g,13.1 mmol) was dissolved in dichloromethane (45 mL) under ice bath and trifluoroacetic acid (15 mL) was added. The reaction solution was slowly returned to room temperature, stirred for 2 hours, and the solvent was dried by spinning to obtain a crude product. The crude product was isolated and purified by C18 reverse phase chromatography to give 5- (6-fluoro-8-hydroxy-2, 3,4, 5-tetrahydro-1H-benzo [ d ] azepin-7-yl) -1,2, 5-thiadiazolidin-3-one 1, 1-dioxide (compound 6) (2.1 g, 51.2% yield). LCMS (ESI)

[M+H]⁺：316.0.¹H NMR(400MHz,DMSO-d₆)δ9.40(brs,1H),7.26(brs,2H),6.59(s,1H),3.93(s,2H),3.23-3.12(m,4H),3.03-2.91(m,4H).

Example 7

Step 1 Compound 6 (1.1 g,3.49 mmol) was dissolved in isopropanol (20 mL) under nitrogen, and acetaldehyde (3.5 mL,17.44 mmol) and glacial acetic acid (0.1 mL) were added. The reaction mixture was stirred at 40 ℃ for 1 hour. Sodium cyanoborohydride (2.4 g,34.88 mmol) was added in portions and the reaction stirred at 40 ℃ for a further 12 hours. The mixture was cooled to room temperature, quenched by addition of ice water (10 mL), and the reaction mixture was concentrated under reduced pressure to give the crude product. The crude product was purified by C18 reverse phase chromatography to give 5- (3-ethyl-6-fluoro-8-hydroxy-2, 3,4, 5-tetrahydro-1H-benzo [ d ] azepin-7-yl) -1,2, 5-thiadiazolidin-3-one 1, 1-dioxide (compound 7) (800 mg, yield 66.8％).LCMS(ESI)[M+H]⁺：344.1.¹H NMR(400MHz,DMSO-d6)δ9.64-9.09(m,2H),6.59(s,1H),3.92(s,2H),3.70-3.45(m,1H),3.26-2.64(m,9H),1.21(t,J＝6.9Hz,3H).

Test example 1 PTPN2 enzymatic inhibition assay

Using 6, 8-difluoro-4-methylumbelliferyl methyl phosphate (DiFMUP) as substrate, 5. Mu.L of PTPN2 solution (reaction buffer PTPN2 diluted to 0.2nM:50mM Tris-HCl, pH 7.2,50mM NaCl,0.01% in 384 well plate)

Triton X-100,1mM DTT) was incubated with compound or DMSO (0.5% v/v) for 10 minutes at room temperature. After the reaction was started by adding 5. Mu.L of DiFMUP (10. Mu.M) and incubating at room temperature for 30min, the fluorescence of the reaction final solution was measured on a CLARIO Star Plusacu enzyme-labeled instrument (BMG) (excitation wavelength 360nm, emission wavelength 460 nm). The experiment was performed with double wells. The low control (DMSO and blank reaction buffer) was set to 100% inhibition, the high control (DMSO and PTPN2 enzyme reaction solution) was set to 0% inhibition. Inhibition was calculated as 100 x (average high control-compound wells)/(average high control-average low control). The IC ₅₀ values were determined using XLfit fit nonlinear regression equations.

TABLE 1 PTPN2 enzymatic Activity (IC ₅₀)

Numbering of compounds	IC50(nM)
		Compound 1	14.9
Compound 2	6.7
		Compound 3	6.5
Compound 4	4.5
		Compound 5	7.2
Compound 6	21.6

Test example 2 Hela p-STAT1 activation detection

Hela cells were seeded in a clear 96-well cell culture plate, placed in a carbon dioxide incubator, after 37-degree overnight incubation, the cell plate was discarded from the supernatant, and 80. Mu.L of basal medium was added. And (3) adding the compound to be tested into a cell culture medium in a gradient dilution way, putting the cell plate back into a carbon dioxide incubator for continuous incubation for 24 hours after uniformly mixing, adding 10 mu L of 50ng/mL IFNgamma, continuously incubating for 30 minutes, and detecting the p-STAT1 by using a Cisbio kit (63 ADK026 PEG) after the incubation is finished. Removing cell supernatant, adding 50 mu L of cell lysate per well, incubating at room temperature for 60 min, taking 16 mu L of cell lysate per well into a new 384-white microplate, adding 2 mu L of Phospho-STAT1 Eu Cryptate antibody diluent and 2 mu L of Phospho-STAT1 d2 anti-body diluent, incubating at room temperature overnight, and reading fluorescent signals (excitation: 320nm, emission: 615nm,665 nm) by using a multi-label analyzer after incubation.

TABLE 2 Hela p-STAT1 Activity (EC ₅₀)

Numbering of compounds	EC50(μM)
		Compound 2	0.76
Compound 7	8.75

While specific embodiments of the invention have been described in detail, those skilled in the art will, in light of all the teachings disclosed, be able to make various modifications and alternatives to the details of the invention and such modifications are within the scope of the invention. The full scope of the invention is given by the appended claims and any equivalents thereof.

Claims (10)

1. A compound of formula I or a pharmaceutically acceptable salt thereof, wherein ring A is selected from an 8-membered heterocyclic group or a cyclooctyl group; ^R1 is selected from _-C1-6 alkyl, _C3-6 cycloalkyl, 5-6 membered heterocyclyl, _-C1-6 haloalkyl, _-C1-6 hydroxyalkyl, _-C1-6 aminoalkyl, _-C1-6 alkyl- _C3-6 cycloalkyl or _-C1-6 alkyl-5-6 membered heterocyclyl; n is selected from 0 or 1. 2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, wherein the compound represented by formula I is the compound represented by formula II, R ^1a is selected from H or R ¹ . 3. The compound or pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein ^R1 is selected from _-C1-3 alkyl, _C3-6 cycloalkyl or _-C1-3 alkyl- _C3-6 cycloalkyl. 4. The compound or pharmaceutically acceptable salt thereof according to claim 1 or 2, wherein ^R1 is selected from _-C1-3 alkyl, cyclopropyl or _-C1-3 alkyl-cyclopropyl. 5. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein the structure of the compound is as follows: 6. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein the structure of the compound is shown as one of the following: 7. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, wherein the structure of the compound is shown as one of the following: 8. A pharmaceutical composition, wherein the pharmaceutical composition comprises the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier. 9. Use of the compound according to any one of claims 1 to 7 or a pharmaceutically acceptable salt thereof or the pharmaceutical composition according to claim 8 in the preparation of a drug for treating a disease mediated by PTPN2. 10. The use according to claim 9, wherein the PTPN2-mediated disease is selected from cancer.