OXA-AZASPIRO DERIVATIVE, AND PREPARATION METHOD THEREFOR AND PHARMACEUTICAL USE THEREOF

Abstract

Disclosed are an oxa-azasipro derivative, a preparation method therefor and a pharmaceutical use thereof. In particular, disclosed are an oxa-azasipro derivative as represented by general formula (I), a preparation method therefor, a pharmaceutical composition containing the derivative, and the use thereof as a therapeutic agent, especially the use thereof as a PI3Kδ inhibitor and the use thereof in the preparation of a drug for treating diseases or conditions improved by means of inhibiting PI3Kδ.

##STR00001##

Claims

1. A compound of general formula (I) or a tautomer, racemate, enantiomer or diastereomer thereof or a mixture thereof, or a pharmaceutically acceptable salt thereof: ##STR00050## wherein: R.sup.a, R.sup.b, R.sup.c, R.sup.d, R.sup.e and R.sup.f are identical or different and are each independently selected from the group consisting of a hydrogen atom, alkyl, halogen, alkoxy, haloalkoxy, cyano, hydroxy, hydroxyalkyl, —(CH.sub.2).sub.sNR.sup.7R.sup.8, cycloalkyl, cycloalkylalkyl, cycloalkyloxy, heterocyclyl, heterocyclylalkyl, heterocyclyloxy, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, haloalkyl, cyano, nitro, —(CH.sub.2).sub.yNR.sup.gR.sup.h and —OR.sup.9; R.sup.0 and R.sup.1, together with the carbon atom to which they are attached, form a spiro ring on the heterocycle to which they are attached, the spiro ring being optionally substituted with one or more R's; the one or more R's are identical or different and are each independently selected from the group consisting of a hydrogen atom, alkyl, halogen, alkoxy, haloalkoxy, cyano, hydroxy, hydroxyalkyl, —(CH.sub.2).sub.sNR.sup.7R.sup.8 and nitro; R.sup.5 is selected from the group consisting of a hydrogen atom, alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, haloalkyl, nitro, cyano, hydroxyalkyl, —(CH.sub.2).sub.yNR.sup.gR.sup.h, —OR.sup.9, —COR.sup.9, —COOR.sup.9, —OS(O).sub.tR.sup.9, —S(O).sub.tR.sup.9, —NR.sup.6COR.sup.9, —NR.sup.6SO.sub.2R.sup.9 and R; R is selected from the group consisting of cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylalkyl, heterocyclylalkyl, arylalkyl and heteroarylalkyl, and the cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylalkyl, heterocyclylalkyl, arylalkyl and heteroarylalkyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, haloalkyl, nitro, cyano, hydroxyalkyl, —(CH.sub.2).sub.yNR.sup.gR.sup.h, —OR.sup.9, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl and heteroaryl; R.sup.2 and R.sup.4 are identical or different and are each independently selected from the group consisting of a hydrogen atom, halogen, alkyl, haloalkyl, alkoxy, cycloalkyl, aryl, heterocyclyl, heteroaryl, cycloalkylalkyl, arylalkyl, heterocyclylalkyl and heteroarylalkyl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of alkyl, haloalkyl, halogen, cyano, nitro, —(CH.sub.2).sub.yNR.sup.gR.sup.h, —OR.sup.9, —COR.sup.9, —COOR.sup.9, —OS(O).sub.tR.sup.9, —S(O).sub.tR.sup.9, —NR.sup.6COR.sup.9 and —NR.sup.6SO.sub.2R.sup.9; each R.sup.3 is identical or different and is independently selected from the group consisting of a hydrogen atom, halogen, alkyl, haloalkyl, cyano, nitro, —(CH.sub.2).sub.sNR.sup.7R.sup.8, —OR.sup.i, —COR.sup.i, —COOR.sup.i, —OS(O).sub.xR.sup.i, —S(O).sub.xR.sup.i, —NR.sup.6COR.sup.i, —NR.sup.6SO.sub.2R.sup.i, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of alkyl, haloalkyl, halogen, cyano, nitro, —(CH.sub.2).sub.yNR.sup.gR.sup.h, —OR.sup.9, —COR.sup.9, —COOR.sup.9, —OS(O).sub.tR.sup.9, —S(O).sub.tR.sup.9, —NR.sup.6COR.sup.9 and —NR.sup.6SO.sub.2R.sup.9; or two adjacent R.sup.3, together with the carbon atom to which they are attached, form a cycloalkyl, heterocyclyl, aryl or heteroaryl, wherein the cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of a hydrogen atom, alkyl, halogen, haloalkyl, alkoxy, haloalkoxy, cyano, amino, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.6 is selected from the group consisting of a hydrogen atom, alkyl, cycloalkyl and aryl, wherein the alkyl, cycloalkyl and aryl are each independently optionally substituted with one or more substituents selected from the group consisting of alkyl, alkoxy, oxo, halogen, amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.7, R.sup.8, R.sup.g and R.sup.h are identical or different and are each independently selected from the group consisting of a hydrogen atom, alkyl, haloalkyl, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; or R.sup.7 and R.sup.8, or R.sup.g and R.sup.h, together with the nitrogen atom to which they are attached, form a heterocyclyl, wherein the heterocyclyl is optionally substituted with one or more substituents selected from the group consisting of alkyl, alkoxy, oxo, halogen, amino, cyano, nitro, hydroxy, hydroxyalkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl; R.sup.9 and R.sup.i are identical or different and are each independently selected from the group consisting of a hydrogen atom, halogen, alkyl, haloalkyl, hydroxyalkyl, alkoxy, haloalkoxy, —(CH.sub.2).sub.sNR.sup.7R.sup.8, cycloalkyl, heterocyclyl, aryl and heteroaryl; wherein the alkyl, cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, alkoxy, haloalkyl, hydroxy, hydroxyalkyl, cyano, amino, nitro, cycloalkyl and heterocyclyl; n is 1 or 2; q is 0, 1, 2, 3 or 4; s and y are identical or different and are each independently selected from the group consisting of 0, 1, 2, 3, 4 and 5; and t and x are identical or different and are each independently selected from the group consisting of 0, 1 and 2.

2. The compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sup.0 and R.sup.1, together with the carbon to which they are attached, form a spiro 3-6 membered ring on the heterocycle to which they are attached.

3. The compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, being a compound of general formula (II) or a tautomer, racemate, enantiomer or diastereomer thereof or a mixture thereof, or a pharmaceutically acceptable salt thereof: ##STR00051## wherein: m is 0, 1, 2 or 3; R′, R.sup.a-R.sup.f, R.sup.2-R.sup.5 and q are as defined in claim 1.

4. The compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sup.5 is aryl or heteroaryl, and the aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, haloalkyl, nitro, cyano, hydroxyalkyl, —(CH.sub.2).sub.yNR.sup.gR.sup.h, —OR.sup.9, —COR.sup.9, —COOR.sup.9, —OS(O).sub.tR.sup.9, —S(O).sub.tR.sup.9, —NR.sup.6COR.sup.9, —NR.sup.6SO.sub.2R.sup.9 and R; R is selected from the group consisting of cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylalkyl, heterocyclylalkyl, arylalkyl and heteroarylalkyl, and the cycloalkyl, heterocyclyl, aryl, heteroaryl, cycloalkylalkyl, heterocyclylalkyl, arylalkyl and heteroarylalkyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl, haloalkyl and —OR.sup.9; R.sup.6, R.sup.9, R.sup.g, R.sup.h, y and t are as defined in claim 1.

5. The compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sup.5 is aryl or heteroaryl, and the aryl and heteroaryl are each independently optionally substituted with R, wherein R is selected from the group consisting of cycloalkylalkyl, heterocyclylalkyl, arylalkyl and heteroarylalkyl, and the cycloalkylalkyl, heterocyclylalkyl, arylalkyl and heteroarylalkyl are each independently optionally substituted with one or more substituents selected from the group consisting of halogen, alkyl and haloalkyl.

6. The compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 3, being a compound of general formula (III) or a tautomer, racemate, enantiomer or diastereomer thereof or a mixture thereof, or a pharmaceutically acceptable salt thereof: ##STR00052## wherein: each R.sup.10 is identical or different and is independently selected from the group consisting of a hydrogen atom, halogen, alkyl, haloalkyl, nitro, cyano, hydroxyalkyl, —(CH.sub.2).sub.sNR.sup.7R.sup.8, —OR.sup.9, cycloalkyl, heterocyclyl, aryl and heteroaryl, wherein the cycloalkyl, heterocyclyl, aryl and heteroaryl are each independently optionally substituted with one or more substituents selected from the group consisting of alkyl, haloalkyl, alkoxy, haloalkoxy, halogen, cyano, nitro and —(CH.sub.2).sub.yNR.sup.gR.sup.h; each R.sup.11 is identical or different and is independently selected from the group consisting of a hydrogen atom, halogen, alkyl, haloalkyl, alkoxy, haloalkoxy, hydroxy, hydroxyalkyl, cyano, nitro, —(CH.sub.2).sub.yNR.sup.gR.sup.h, cycloalkyl, cycloalkyloxy and cycloalkylalkyl; each R.sup.12 is identical or different and is each independently selected from the group consisting of a hydrogen atom, halogen, alkyl, haloalkyl, nitro, cyano, hydroxyalkyl, —(CH.sub.2).sub.yNR.sup.gR.sup.h, —OR.sup.9, cycloalkyl, cycloalkylalkyl, heterocyclyl, heterocyclylalkyl, aryl and heteroaryl; provided that when u is greater than or equal to 2, two R.sup.12 form a spiro or bridged ring system on a morpholine ring; w is 0, 1, 2, 3 or 4; u is 0, 1, 2, 3, 4, 5 or 6; R′, R.sup.a-R.sup.h, R.sup.2-R.sup.4, R.sup.7-R.sup.9, s, m, y and q are as defined in claim 3.

7. The compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sup.2 and R.sup.4 are identical or different and are each independently selected from the group consisting of a hydrogen atom, halogen and C.sub.1-6 alkyl.

8. The compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, wherein each R.sup.3 is identical or different and is independently selected from the group consisting of a hydrogen atom, halogen, C.sub.1-6 haloalkyl, C.sub.1-6 alkoxy and C.sub.1-6 alkyl.

9. The compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 6, wherein each R.sup.10 is identical or different and is independently selected from the group consisting of a hydrogen atom, halogen and C.sub.1-6 alkyl.

10. The compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 6, wherein each R.sup.11 is identical or different and is independently selected from the group consisting of a hydrogen atom, halogen and C.sub.1-6 alkyl.

11. The compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 6, wherein each R.sup.12 is identical or different and is independently selected from the group consisting of a hydrogen atom, halogen and C.sub.1-6 alkyl.

12. The compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, wherein R.sup.a-R.sup.f are each independently a hydrogen atom.

13. A compound selected from the group consisting of the following compounds: ##STR00053## ##STR00054## ##STR00055## or a tautomer, racemate, enantiomer or diastereomer thereof or a mixture thereof, or a pharmaceutically acceptable salt thereof.

14. A method for preparing the compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, comprising the following step: ##STR00056## subjecting a compound of general formula (IA) or a tautomer, racemate, enantiomer or diastereomer thereof or a mixture thereof, or a pharmaceutically acceptable salt thereof to a reaction with a compound of general formula (IB) or a pharmaceutically acceptable salt thereof to give the compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof, wherein R.sup.0-R.sup.5, R.sup.aR.sup.f, n and q are as defined in claim 1.

15. A pharmaceutical composition; comprising the compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1, and one or more pharmaceutically acceptable carriers, diluents or excipients.

16. A method for inhibiting PI3Kδ in a subject in need thereof, the method comprising administering to subject an effective amount of the compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1.

17. A method for treating and/or preventing an inflammatory disease, an autoimmune disease, a cancer or a related disease in a subject in need thereof, the method comprising administering to the subject an effective amount of the compound of general formula (I) or the tautomer, racemate, enantiomer or diastereomer thereof or the mixture thereof, or the pharmaceutically acceptable salt thereof according to claim 1.

18. The method according to claim 17, wherein the cancer is selected from the group consisting of melanoma, skin cancer, liver cancer, kidney cancer, lung cancer, nasopharyngeal cancer, gastric cancer, esophageal cancer, colorectal cancer, gallbladder cancer, bile duct cancer, chorionic epithelioma, pancreatic cancer, polycythemia vera, pediatric tumors, cervical cancer, ovarian cancer, breast cancer, bladder cancer, urothelial cancer, ureteral tumor, prostate cancer, seminoma, testicular tumor, leukemia, head and neck tumor, endometrial cancer, thyroid cancer, lymphoma, sarcoma, osteoma, neuroturbo chargeoma, neuroblastoma, neuroendocrine carcinoma, brain tumor, CNS cancer, myeloma, astrocytoma, glioblastoma and glioma.

19. A method for inhibiting PI3Kδ in a subject in need thereof, the method comprising administering to the subject an effective amount of the pharmaceutical composition according to claim 15.

20. A method for treating and/or preventing an inflammatory disease, autoimmune disease, a cancer or a related disease in a subject in need thereof, the method comprising administering to the subject an effective amount of the pharmaceutical composition according to claim 15.

Description

DETAILED DESCRIPTION

[0190] The following examples further illustrate the present disclosure, but the present disclosure is not limited thereto.

EXAMPLES

[0191] The structure of the compound is determined by nuclear magnetic resonance (NMR) spectroscopy and/or mass spectrometry (MS). NMR shift (δ) is given in a unit of 10.sup.−6 (ppm). NMR spectra were measured using a Bruker AVANCE NEO 500M nuclear magnetic resonance instrument, with deuterated dimethyl sulfoxide (DMSO-d.sub.6), deuterated chloroform (CDCl.sub.3) and deuterated methanol (CD30D) as determination solvents and tetramethylsilane (TMS) as internal standard.

[0192] Mass spectra were measured using Agilent 1200/1290 DAD-6110/6120 Quadrupole MS liquid chromatography-mass spectrometry system (manufacturer: Agilent; MS model: 6110/6120 Quadrupole MS), Waters ACQuity UPLC-QD/SQD (manufacturer: Waters, MS model: Waters ACQuity Qda Detector/Waters SQ Detector) and THERMO Ultimate 3000-Q Exactive (manufacturer: THERMO, MS model: THERMO Q Exactive).

[0193] High performance liquid chromatography (HPLC) analysis is performed using Agilent HPLC 1200DAD, Agilent HPLC 1200VWD and Waters HPLC e2695-2489 high performance liquid chromatographs.

[0194] Chiral HPLC analysis is performed using an Agilent 1260 DAD high performance liquid chromatograph.

[0195] High performance liquid preparative chromatography is performed using Waters 2545-2767, Waters 2767-SQ Detecor2, Shimadzu LC-20AP and Gilson GX-281 preparative chromatographs.

[0196] Chiral preparative HPLC is performed using a Shimadzu LC-20AP preparative chromatograph.

[0197] CombiFlash rapid preparation instrument used is CombiFlash R.sup.f200 (TELEDYNE ISCO).

[0198] Huanghai HSGF254 or Qingdao GF254 silica gel plates of specifications 0.15 mm to 0.2 mm are adopted for thin layer chromatography (TLC) analysis and 0.4 mm to 0.5 mm for TLC separation and purification.

[0199] The silica gel column chromatography generally used 200 to 300-mesh silica gel (Huanghai, Yantai) as the carrier.

[0200] The mean inhibition of kinase and the IC.sub.50 value are determined using a NovoStar microplate reader (BMG, Germany).

[0201] Known starting materials described herein may be synthesized using or according to methods known in the art, or may be purchased from ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc., Chembee Chemicals, and other companies.

[0202] In the examples, the reactions can be performed in an argon atmosphere or a nitrogen atmosphere unless otherwise specified.

[0203] The argon atmosphere or nitrogen atmosphere means that the reaction flask is connected to a balloon containing about 1 L of argon or nitrogen.

[0204] The hydrogen atmosphere means that the reaction flask is connected to a balloon containing about 1 L of hydrogen.

[0205] Parr 3916EKX hydrogenator, Qinglan QL-500 hydrogenator or HC2-SS hydrogenator was used in the pressurized hydrogenation reactions.

[0206] The hydrogenation reactions usually involve 3 cycles of vacuumization and hydrogen purge.

[0207] A CEM Discover-S 908860 microwave reactor is used in the microwave reactions.

[0208] In the examples, a solution refers to an aqueous solution unless otherwise specified.

[0209] In the examples, the reaction temperature is room temperature, i.e., 20° C. to 30° C., unless otherwise specified.

[0210] The monitoring of the reaction progress in the examples was conducted by thin layer chromatography (TLC). The developing solvent for reactions, the eluent system for column chromatography purification and the developing solvent system for thin layer chromatography included: A: dichloromethane/methanol system. The volume ratio of the solvents was adjusted according to the polarity of the compound, or by adding a small amount of basic or acidic reagents such as triethylamine and acetic acid.

Example 1

(6-fluoro-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 1

[0211] ##STR00035## ##STR00036##

Step 1

Tert-butyl 1-(4-(morpholinomethyl)phenyl)hydrazine-1-carboxylate 2b

[0212] Compound 4-(4-iodobenzyl)morpholine 2a (51 g, 168.24 mmol, prepared according to the method disclosed in Example 17.1 (page 59) in the specification of the patent application WO200832191A2) and tert-butyl carbazate (23.347 g, 176.66 mmol, Accela ChemBio) were dissolved in dimethyl sulfoxide (400 mL) under argon atmosphere, and the mixture was stirred for 10 min, followed by addition of cuprous iodide (1.603 g, 8.42 mmol). The resulting mixture was warmed to 50° C. and stirred for 17 h. Water (400 mL) was added, and the aqueous phase was extracted with ethyl acetate (300 mL×6). The organic phases were combined and concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 2b (51 g, yield: 98.6%).

[0213] MS m/z (ESI): 308.1 [M+1].

Step 2

Hydrochloride of 4-(4-hydrazinobenzyl)morpholine 2c

[0214] Compound 2b (51 g, 165.91 mmol) was dissolved in methanol (80 mL) at 0° C., and a solution of hydrogen chloride in 1,4-dioxane (350 mL, 4.0 M, Infinity Scientific) was added dropwise. The mixture was naturally warmed to room temperature and stirred for 17 h. Then the mixture was concentrated under reduced pressure to give crude hydrochloride of the title compound 2c (45.4 g), which was directly used in the next step without purification.

Step 3

3-((2-fluorophenyl)thio)propanoic acid 1e

[0215] 2-fluorobenzenethiol 1d (50 g, 390.11 mmol, Accela ChemBio) and potassium carbonate (70 g, 507.14 mmol, Sinopharm) were dissolved in N,N-dimethylformamide (500 mL), and the mixture was stirred at 60° C. for 1 h, followed by addition of 3-bromopropionic acid (65.6 g, 429.15 mmol, Accela ChemBio). The resulting mixture was reacted at 60° C. for 3 h. After the reaction was completed, water (1000 mL) was added to the reaction solution, and ethyl acetate (300 mL×2) was added for extraction. The aqueous phase was adjusted to pH of about 3 with concentrated hydrochloric acid and extracted with ethyl acetate (400 mL×3). The organic phases were combined, washed successively with water (400 mL×3) and saturated brine (400 mL×2), dried over anhydrous sodium sulfate for 15 min, filtered and concentrated under reduced pressure to give crude title compound 1e (70 g), which was directly used in the next step without purification.

[0216] MS m/z (ESI): 198.9 [M−1].

Step 4

8-fluorothiochroman-4-one 1f

[0217] Compound 1e (70 g, 356.65 mmol) was dissolved in concentrated sulfuric acid (200 mL), and the mixture was stirred at 0° C. for 3 h. After the reaction was completed, the reaction solution was poured into ice water (1000 mL), and ethyl acetate (400 mL×3) was added for extraction. The organic phases were combined, washed with saturated brine (400 mL×2), dried over anhydrous sodium sulfate for 15 min and filtered, and the filtrate was concentrated under reduced pressure to give crude title compound if (30 g), which was directly used in the next step without purification.

Step 5

Ethyl 2-(8-fluoro-4-oxothiochroman-3-yl)-2-oxoacetate 1g

[0218] Sodium ethoxide (52 g, 152.59 mmol, 20% w/w ethanol solution, Adamas) was added to a 500 mL three-neck flask, and diethyl oxalate (16.7 g, 114.47 mmol, dissolved in 100 mL of toluene, Accela ChemBio) was slowly added dropwise at 0° C., followed by addition of compound if (13.9 g, 76.28 mmol, dissolved in 100 mL of toluene). The reaction solution was concentrated under reduced pressure, water (400 mL) was added to the residue, and dichloromethane (200 mL×2) was added for extraction. The aqueous phase was adjusted to pH of about 2 with 5 M hydrochloric acid solution and extracted with ethyl acetate (250 mL×3). The organic phases were combined, washed with saturated brine (200 mL×2), dried over anhydrous sodium sulfate for 15 min and filtered, and the filtrate was concentrated under reduced pressure to give crude title compound 1g (22 g), which was directly used in the next step without purification.

[0219] MS m/z (ESI): 280.9 [M−1].

Step 6

Ethyl 2-(8-fluoro-1,1-dioxido-4-oxothiochroman-3-yl)-2-oxoacetate 1h

[0220] Compound 1g (22 g, 77.93 mmol) was dissolved in dichloromethane (250 mL), and 3-chloroperoxybenzoic acid (34.8 g, 171.46 mmol, Ourchem) was added in portions under an ice bath. The mixture was stirred at room temperature for 17 h. Then the mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 1h (24.5 g, yield: 100%).

[0221] MS m/z (ESI): 313.0 [M−1].

Step 7

Ethyl 6-fluoro-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylate 5,5-dioxide 1i

[0222] Compound 1h (9.4 g, 29.90 mmol), the hydrochloride of compound 2c (6.8 g, 75%) and glacial acetic acid (3.6 g, 59.94 mmol, Hushi) were dissolved in absolute ethanol (200 mL), and the mixture was heated to reflux and stirred for 3 h. Saturated sodium bicarbonate solution (300 mL) was added, and the mixed solution was extracted with ethyl acetate (250 mL×3). The organic phases were combined and concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 1i (7.5 g, yield: 51.6%).

[0223] MS m/z (ESI): 486.1 [M+1].

Step 8

6-fluoro-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylic acid 5,5-dioxide 1a

[0224] Compound 1i (14 g, 29.07 mmol) was dissolved in tetrahydrofuran (150 mL), and aqueous sodium hydroxide solution (58.2 mL, 2 M) was added. The mixture was stirred for 4 h. Concentrated hydrochloric acid solution was added to adjust the pH to about 3, and the resulting mixture was concentrated under reduced pressure to give crude title compound 1a (21.2 g), which was directly used in the next step without purification.

[0225] MS m/z (ESI): 458.0 [M+1].

Step 9

(6-fluoro-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 1

[0226] Compound 1a (100 mg, 218.59 mmol), 4-oxa-7-azaspiro[2.5]octane hydrochloride (33 mg, 220.77 mmol, Jiangsu Aikon), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (251 mg, 651.77 mmol), 1-hydroxybenzotriazole (99 mg, 655.77 mmol), N,N-diisopropylethylamine (110 mg, 1.09 mmol) and 4-dimethylaminopyridine (53 mg, 437.18 mmol) were dissolved in dichloromethane (30 mL), and the mixture was stirred at room temperature for 16 h. Water (50 mL) was added, and a mixed solvent (60 mL) of dichloromethane and methanol (v:v=10:1) was added for extraction. The organic phases were combined, washed successively with water (60 mL) and saturated brine (60 mL), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title product 1 (60 mg, yield: 49.6%).

[0227] MS m/z (ESI): 553.3 [M+1].

[0228] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 7.62-7.44 (m, 6H), 6.66 (d, 1H), 4.99 (d, 2H), 4.07 (t, 1H), 3.90 (s, 1H), 3.76-3.59 (m, 9H), 3.32 (s, 1H), 2.42 (s, 4H), 0.73-0.60 (m, 4H).

Example 2

(6-chloro-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 2

[0229] ##STR00037##

Step 1

Tert-butyl 1-(4-(morpholinomethyl)phenyl)hydrazine-1-carboxylate 2b

[0230] Compound 4-(4-iodobenzyl)morpholine 2a (51 g, 168.24 mmol, prepared according to the method disclosed in Example 17.1 (page 59) in the specification of the patent application WO200832191A2) and tert-butyl carbazate (23.347 g, 176.66 mmol, Accela ChemBio) were dissolved in dimethyl sulfoxide (400 mL) under argon atmosphere, and the mixture was stirred for 10 min, followed by addition of cuprous iodide (1.603 g, 8.42 mmol). The resulting mixture was warmed to 50° C. and stirred for 17 h. Water (400 mL) was added, and the aqueous phase was extracted with ethyl acetate (300 mL×6). The organic phases were combined and concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 2b (51 g, yield: 98.6%).

[0231] MS m/z (ESI): 308.1 [M+1].

Step 2

Hydrochloride of 4-(4-hydrazinobenzyl)morpholine 2c

[0232] Compound 2b (51 g, 165.91 mmol) was dissolved in methanol (80 mL) at 0° C., and a solution of hydrogen chloride in 1,4-dioxane (350 mL, 4.0 M, Infinity Scientific) was added dropwise. The mixture was naturally warmed to room temperature and stirred for 17 h. Then the mixture was concentrated under reduced pressure to give crude hydrochloride of the title compound 2c (45.4 g), which was directly used in the next step without purification.

Step 3

Ethyl 2-(8-chloro-1,1-dioxido-4-oxothiochroman-3-yl)-2-oxoacetate 2e

[0233] Compound ethyl 2-(8-chloro-4-oxothiochroman-3-yl)-2-oxoacetate 2d (25.7 g, 86.03 mmol, prepared according to the disclosed method for intermediate E4 (page 169) in the specification of the patent “CN102695710B”) was dissolved in dichloromethane (250 mL), and 3-chloroperoxybenzoic acid (43.664 g, 215.07 mmol) was added in portions under an ice bath. The mixture was stirred at room temperature for 17 h. Then the mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 2e (28.323 g, yield: 99.5%).

[0234] MS m/z (ESI): 330.9 [M+1].

Step 4

Ethyl 6-chloro-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylate 5,5-dioxide 2f

[0235] Compound 2e (10 g, 30.24 mmol), the hydrochloride of compound 2c (9.192 g, 75%) and glacial acetic acid (3.632 g, 60.48 mmol, Hushi) were dissolved in absolute ethanol (300 mL), and the mixture was heated to reflux and stirred for 3 h. Saturated sodium bicarbonate solution (300 mL) was added, and the mixed solution was extracted with ethyl acetate (250 mL×3). The organic phases were combined and concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 2f (10 g, yield: 65.9%).

[0236] MS m/z (ESI): 502.0 [M+1].

Step 5

6-chloro-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylic acid 5,5-dioxide 2g

[0237] Compound 2f (10 g, 19.92 mmol) was dissolved in tetrahydrofuran (150 mL), and aqueous sodium hydroxide solution (39.8 mL, 2.5 M) was added. The mixture was stirred for 4 h. Concentrated hydrochloric acid solution was added to adjust the pH to about 3, and the resulting mixture was concentrated under reduced pressure to give crude title compound 2g (17.028 g), which was directly used in the next step without purification.

[0238] MS m/z (ESI): 474.0 [M+1].

Step 6

(6-chloro-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 2

[0239] Compound 2g (544 mg, 633.61 μmol, 55.2%), 4-oxa-7-azaspiro[2.5]octane hydrochloride (97 mg, 648.32 μmol, PharmaBlock), HATU (290 mg, 762.70 μmol) and N,N-diisopropylethylamine (411 mg, 3.18 mmol) were dissolved in N,N-dimethylformamide (60 mL), and the mixture was stirred at room temperature for 17 h. Saturated sodium bicarbonate solution (50 mL) was added, and the aqueous phase was extracted with ethyl acetate (50 mL×3). The organic phases were combined and concentrated under reduced pressure, and the resulting residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title product 2 (108 mg, yield: 30.0%).

[0240] MS m/z (ESI): 569.0 [M+1].

[0241] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 7.66-7.64 (m, 1H), 7.54-7.40 (m, 5H), 6.83-6.81 (m, 1H), 5.02-5.00 (m, 2H), 4.08 (s, 1H), 3.92 (s, 1H), 3.75-3.58 (m, 10H), 2.46-2.41 (m, 4H), 0.73-0.61 (m, 4H).

Example 3

(9-methoxy-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 3

[0242] ##STR00038##

Step 1

5-methoxythiochroman-4-one 3b

[0243] 3-(3-methoxyphenylthio)propionic acid 3a (12 g, 56.53 mmol, prepared according to the method disclosed in “Organic Letters, 2020, 22(3), 1155-1159”) and sulfuric acid (40 mL) were added into a 100 mL one-neck flask, and the mixture was stirred at room temperature for 3 h. The reaction solution was poured into ice water (100 mL), and ethyl acetate (100 mL×3) was added for extraction. The organic phase was washed with saturated brine (100 mL×2), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the target product 3b (300 mg, yield: 2.73%).

Step 2

Ethyl 2-(5-methoxy-4-oxothiochroman-3-yl)-2-oxoacetate 3c

[0244] Sodium ethoxide (1.44 g, 4.23 mmol, 20% w/w ethanol solution) was dissolved in toluene (20 mL), and the mixture was cooled to 0° C. A solution of diethyl oxalate (463 mg, 3.166 mmol) in toluene (20 mL) was added dropwise, followed by the addition of compound 3b (410 mg, 2.11 mmol). The resulting mixture was reacted at room temperature for 17 h. The reaction solution was concentrated under reduced pressure, water (100 mL) was added to the residue, and dichloromethane (50 mL) was added for extraction. The aqueous phase was adjusted to pH of about 2 with 5 M hydrochloric acid solution and extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated brine (20 mL×2), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the title compound 3c (900 mg), which was directly used in the next step without purification.

Step 3

Ethyl 2-(5-methoxy-1,1-dioxido-4-oxothiochroman-3-yl)-2-oxoacetate 3d

[0245] Crude compound 3c (900 mg, 3.058 mmol) was dissolved in dichloromethane (30 mL), and 3-chloroperoxybenzoic acid (1.2 g, 6.95 mmol) was added. The mixture was stirred at room temperature for 17 h. Then the mixture was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system B to give the title compound 3d (550 mg, yield: 55.1%).

Step 4

Ethyl 9-methoxy-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylate 5,5-dioxide 3e

[0246] Compound 3d (550 mg, 1.68 mmol) was dissolved in ethanol (30 mL), and the hydrochloride of compound 2c (384 mg) and glacial acetic acid (203 mg, 3.3804 mmol) were added. The mixture was stirred at 90° C. for 2 h. Then the mixture was concentrated under reduced pressure, slurried with ethanol and filtered, and the filter cake was dried to give the title product 3e (700 mg, yield: 83.4%).

Step 5

9-methoxy-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylic acid 5,5-dioxide 3f

[0247] Compound 3e (10.2 g, 20.5 mmol) was dissolved in tetrahydrofuran (50 mL), and aqueous sodium hydroxide solution (3 M, 2.8 mL) was added. The mixture was stirred at room temperature for 4 h. The reaction solution was adjusted to pH of about 2 with 5.0 M hydrochloric acid solution and concentrated under reduced pressure to give crude title product 3f (1 g, purity: 60%), which was directly used in the next step without purification.

Step 6

(9-methoxy-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 3

[0248] Crude compound 3f (300 mg, 332.26 μmol, purity: 60%), 4-oxa-7-azaspiro[2.5]octane hydrochloride (50 mg, 334.18 μmol, PharmaBlock), HATU (152 mg, 399.76 μmol) and N,N-diisopropylethylamine (215 mg, 1.66 mmol) were dissolved in N,N-dimethylformamide (60 mL), and the mixture was stirred at room temperature for 17 h. Saturated sodium bicarbonate solution (50 mL) was added, and the aqueous phase was extracted with ethyl acetate (50 mL×3). The organic phases were combined and concentrated under reduced pressure, and the resulting residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title product 3 (40 mg, yield: 21.3%).

[0249] MS m/z (ESI): 565.0 [M+1].

[0250] .sup.1H NMR (500 MHz, CDCl.sub.3) δ 7.77-7.75 (m, 1H), 7.59-7.56 (m, 1H), 7.42-7.39 (m, 2H), 7.35-7.33 (m, 1H), 7.31-7.30 (m, 1H), 7.03-7.01 (m, 1H), 4.73-4.71 (m, 2H), 4.33-4.32 (m, 1H), 4.19 (s, 1H), 3.89-3.82 (m, 3H), 3.79-3.74 (m, 5H), 3.57 (s, 2H), 3.13 (s, 3H), 2.50-2.48 (m, 4H), 0.87-0.85 (m, 2H), 0.74-0.72 (m, 2H).

Example 4

(7-methoxy-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 4

[0251] ##STR00039##

Step 1

Ethyl 2-(7-methoxy-4-oxothiochroman-3-yl)-2-oxoacetate 4b

[0252] Sodium ethoxide (19.267 g, 56.62 mmol, 20% w/w ethanol solution) was added into a 500 mL single-neck flask, and a solution of diethyl oxalate (6.207 g, 42.47 mmol) in toluene (300 mL) was added at 0° C., followed by addition of 7-methoxythiochroman-4-one 4a (5.5 g, 28.31 mmol, prepared according to the method disclosed in “Organic Letters, 2020, 22(3), 1155-1159”). The mixture was stirred at room temperature for 17 h. The reaction solution was concentrated under reduced pressure, water (400 mL) was added to the residue, and dichloromethane (200 mL×2) was added for extraction. The aqueous phase was adjusted to pH of about 2 with 5 M hydrochloric acid solution and extracted with ethyl acetate (200 mL×3). The organic phases were combined, washed with saturated brine (200 mL×2), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give crude title compound 4b (8.3 g), which was directly used in the next step without purification.

Step 2

Ethyl 2-(7-methoxy-1,1-dioxido-4-oxothiochroman-3-yl)-2-oxoacetate 4c

[0253] Crude compound 4b (8.3 g, 28.20 mmol) was dissolved in dichloromethane (200 mL), and 3-chloroperoxybenzoic acid (12.166 g, 70.50 mmol) was added in portions under an ice bath. The mixture was stirred at room temperature for 17 h. Then the mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system B to give the title compound 4c (8.8 g, yield: 95.6%).

Step 3

Ethyl 7-methoxy-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylate 5,5-dioxide 4d

[0254] Compound 4c (8.8 g, 26.96 mmol) was dissolved in ethanol (200 mL), and the hydrochloride of 2c (6.7 g) and glacial acetic acid (3.239 g, 53.93 mmol) were added. The mixture was stirred at 90° C. for 2 h. Then the mixture was concentrated under reduced pressure, slurried with ethanol and filtered, and the filter cake was dried to give the title product 4d (10.2 g, yield: 76.0%).

Step 4

7-methoxy-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylic acid 5,5-dioxide 4e

[0255] Compound 4d (10.2 g, 20.5 mmol) was dissolved in tetrahydrofuran (100 mL), and aqueous sodium hydroxide solution (1.0 M, 102.5 mL) was added. The mixture was stirred at room temperature for 4 h. The reaction solution was adjusted to pH of about 2 with 5.0 M hydrochloric acid solution and concentrated under reduced pressure to give the title product 4e (16.3 g, 5 purity: 8.8%).

Step 5

(7-methoxy-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 4

[0256] Compound 4e (300 mg, 332.26 μmol, 58.8%), 4-oxa-7-azaspiro[2.5]octane hydrochloride (50 mg, 334.18 μmol, PharmaBlock), HATU (151 mg, 397.13 μmol) and N,N-diisopropylethylamine (215 mg, 1.66 mmol) were dissolved in N,N-dimethylformamide (5 mL), and the mixture was stirred at room temperature for 17 h. Saturated sodium bicarbonate solution (10 mL) was added, and the aqueous phase was extracted with ethyl acetate (20 mL×3). The organic phases were combined and concentrated under reduced pressure, and the resulting residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title product 4 (47 mg, yield: 25.1%).

[0257] MS m/z (ESI): 565.1 [M+1].

[0258] .sup.1H NMR (500 MHz, CDCl.sub.3) δ 7.62-7.60 (m, 1H), 7.53-7.49 (m, 2H), 7.46-7.40 (m, 2H), 7.07-6.89 (m, 1H), 6.83-6.81 (m, 1H), 4.76-4.75 (m, 2H), 4.31-4.29 (m, 1H), 4.18-4.16 (m, 1H), 3.91-3.88 (m, 5H), 3.83-3.78 (m, 6H), 3.63-3.60 (m, 2H), 2.54-2.51 (m, 4H), 0.87-0.84 (m, 2H), 0.72-0.68 (m, 2H).

Example 5

(6-methoxy-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 5

[0259] ##STR00040##

Step 1

3-((2-methoxyphenyl)thio)propanoic acid 5b

[0260] 2-methoxybenzenethiol 5a (25.0 g, 178.31 mmol, Accela ChemBio) and potassium carbonate (36.9 g, 267.50 mmol, Sinopharm) were dissolved in N,N-dimethylformamide (200 mL). The mixture was stirred at 60° C. for 30 min under nitrogen atmosphere and cooled to room temperature, and 3-bromopropionic acid (28.6 g, 187.28 mmol, Adamas) was added. The resulting mixture was stirred at 60° C. for 3 h under nitrogen atmosphere. Water (1000 mL) was added to the reaction solution, and ethyl acetate (300 mL×2) was added for extraction. The aqueous phase was adjusted to pH of about 3 with concentrated hydrochloric acid and extracted with ethyl acetate (400 mL×2). The organic phases were combined, washed successively with water (400 mL×2) and saturated brine (400 mL×2), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the title product 5b (37 g, yield: 98%).

[0261] MS m/z (ESI): 213.1 [M−1].

Step 2

8-methoxythiochroman-4-one 5c

[0262] Compound 5b (37 g, 174.3 mmol) was dissolved in concentrated sulfuric acid (200 mL), and the mixture was stirred at 0° C. for 2 h. The reaction solution was poured into ice water (1000 mL), and ethyl acetate (300 mL×3) was added for extraction. The organic phase was washed with saturated brine (300 mL×2), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system B to give the title product 5c (1.74 g, yield: 4.3%).

[0263] MS m/z (ESI): 194.9 [M+1].

Step 3

Ethyl 2-(8-methoxy-4-oxothiochroman-3-yl)-2-oxoacetate 5d

[0264] Sodium ethoxide (6.10 g, 17.92 mmol, 20% w/w ethanol solution) was added to a 100 mL three-neck flask, and diethyl oxalate (1.97 g, 13.49 mmol, dissolved in 30 mL of toluene) and compound 5c (1.74 g, 8.95 mmol, dissolved in 30 mL of toluene) were added at 0° C. The mixture was reacted at room temperature for 16 h. The reaction solution was concentrated under reduced pressure, water (80 mL) was added to the residue, and dichloromethane (80 mL×2) was added for extraction. The aqueous phase was adjusted to pH of about 2 with 5 M hydrochloric acid solution and extracted with ethyl acetate (70 mL×3). The organic phases were combined, washed with saturated brine (60 mL×2), dried over anhydrous sodium sulfate for 15 min and filtered, and the filtrate was concentrated under reduced pressure to give the title product 5d (2.6 g, yield: 98.6%).

[0265] MS m/z (ESI): 295.0 [M+1].

Step 4

Ethyl 2-(8-methoxy-1,1-dioxido-4-oxothiochroman-3-yl)-2-oxoacetate 5e

[0266] Compound 5d (2.6 g, 8.83 mmol) was dissolved in dichloromethane (250 mL), and 3-chloroperoxybenzoic acid (3.9 g, 19.47 mmol) was added in portions under an ice bath. The mixture was stirred at room temperature for 17 h. Then the mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 5e (2.8 g, yield: 97.1%).

[0267] MS m/z (ESI): 326.9 [M+1].

Step 5

Ethyl 6-methoxy-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylate 5,5-dioxide 5f

[0268] Compound 5e (1.3 g, 3.98 mmol), the hydrochloride of compound 2c (825.7 mg) and glacial acetic acid (478.4 mg, 7.96 mmol) were dissolved absolute ethanol (60 mL), and the mixture was heated to reflux and stirred for 3 h. Saturated sodium bicarbonate solution (60 mL) was added, and the mixed solution was extracted with ethyl acetate (80 mL×3). The organic phases were combined and concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 5f (1.63 g, yield: 82.2%).

[0269] MS m/z (ESI): 498.0 [M+1].

Step 6

6-methoxy-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylic acid 5,5-dioxide 5g

[0270] Compound 5f (1.63 g, 3.27 mmol) was dissolved in tetrahydrofuran (30 mL), and aqueous sodium hydroxide solution (6.5 mL, 2.5 M) was added. The mixture was stirred for 4 h. Concentrated hydrochloric acid solution was added to adjust the pH to about 3, and the resulting mixture was concentrated under reduced pressure to give crude title compound 5g (2.4 g, yield: 156.0%), which was directly used in the next step without purification.

[0271] MS m/z (ESI): 470.0 [M+1].

Step 7

(6-methoxy-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 5

[0272] Crude compound 5g (330 mg, 421.71 μmol, purity: 60%), 4-oxa-7-azaspiro[2.5]octane hydrochloride (63 mg, 421.74 μmol, Jiangsu Aikon) and HATU (297.6 mg, 1.26 mmol) were dissolved in N,N-dimethylformamide (30 mL), and N,N-diisopropylethylamine (326 mg, 2.53 mmol) was added. The mixture was stirred for 2 h. Water (60 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (80 mL×3). The organic phases were combined, washed with saturated brine (80 mL×3), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 5 (102 mg, yield: 42.8%).

[0273] MS m/z (ESI): 565.1 [M+1].

[0274] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 7.53-7.50 (m, 2H), 7.45-7.37 (m, 3H), 7.25 (d, 1H), 6.39-6.37 (m, 1H), 4.80 (d, 2H), 4.07 (m, 2H), 3.91 (s, 3H), 3.74-3.57 (m, 9H), 3.32 (s, 1H), 2.41 (t, 4H), 0.74-0.59 (m, 4H).

Example 6

(6-methyl-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 6

[0275] ##STR00041##

Step 1

3-(o-tolylthio)propionic acid 6b

[0276] 2-methylbenzenethiol 6a (25.0g, 201.2 mmol, Accela ChemBio) and potassium carbonate (41.7 g, 301.9 mmol) were dissolved in N,N-dimethylformamide (200 mL), and the mixture was stirred at 60° C. for 30 min under nitrogen atmosphere. Then the mixture was cooled to room temperature, followed by addition of 3-bromopropionic acid (32.3 g, 211.4 mmol, Adamas). The resulting mixture was stirred at 60° C. for 3 h under nitrogen atmosphere. Water (1000 mL) was added to the reaction solution, and ethyl acetate (300 mL×2) was added for extraction. The aqueous phase was adjusted to pH of about 3 with concentrated hydrochloric acid and extracted with ethyl acetate (400 mL×2). The organic phases were combined, washed successively with water (400 mL×2) and saturated brine (400 mL×2), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the title product 6b (39 g, yield: 98%).

[0277] MS m/z (ESI): 195.2 [M−1].

Step 2

8-methylthiochroman-4-one 6c

[0278] Compound 6b (39 g, 198.6 mmol) was dissolved in concentrated sulfuric acid (200 mL), and the mixture was stirred at 0° C. for 2 h. The reaction solution was poured into ice water (1000 mL), and ethyl acetate (300 mL×3) was added for extraction. The organic phase was washed with saturated brine (300 mL×2) and dried over anhydrous sodium sulfate, the filtrate was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system B to give the title product 6c (15.5 g, yield: 43%).

[0279] MS m/z (ESI): 178.9 [M+1].

Step 3

Ethyl 2-(8-methyl-4-oxothiochroman-3-yl)-2-oxoacetate 6d

[0280] Sodium ethoxide (59 g, 173.93 mmol, 20% w/w ethanol solution) was added to a 500 mL three-neck flask, and diethyl oxalate (19 g, 130.49 mmol, dissolved in 100 mL of toluene) and compound 6c (15.5 g, 86.9 mmol, dissolved in 100 mL of toluene) were added at 0° C. The mixture was reacted at room temperature for 16 h. The reaction solution was concentrated under reduced pressure, water (400 mL) was added to the residue, and dichloromethane (200 mL×2) was added for extraction. The aqueous phase was adjusted to pH of about 2 with 5 M hydrochloric acid solution and extracted with ethyl acetate (200 mL×3). The organic phases were combined, washed with saturated brine (200 mL×2), dried over anhydrous sodium sulfate for 15 min and filtered, and the filtrate was concentrated under reduced pressure to give the title product 6d (24 g, yield: 99.0%).

[0281] MS m/z (ESI): 279.0 [M+1].

Step 4

Ethyl 2-(8-methyl-1,1-dioxido-4-oxothiochroman-3-yl)-2-oxoacetate 6e

[0282] Compound 6d (24 g, 86.23 mmol) was dissolved in dichloromethane (250 mL), and 3-chloroperoxybenzoic acid (29 g, 172.5 mmol) was added in portions under an ice bath. The mixture was stirred at room temperature for 17 h. Then the mixture was filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 6e (26 g, yield: 97.10%).

[0283] MS m/z (ESI): 310.9 [M+1].

Step 5

Ethyl 6-methyl-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylate 5,5-dioxide 6f

[0284] Compound 6e (15 g, 49.01 mmol), the hydrochloride of compound 2c (10 g) and glacial acetic acid (5.9 g, 98.11 mmol) were dissolved in absolute ethanol (300 mL), and the mixture was heated to reflux and stirred for 3 h. Saturated sodium bicarbonate solution (300 mL) was added, and the mixed solution was extracted with ethyl acetate (250 mL×3). The organic phases were combined and concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 6f (20 g, yield: 86.9%).

[0285] MS m/z (ESI): 482.0 [M+1].

Step 6

6-methyl-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylic acid 5,5-dioxide 6g

[0286] Compound 6f (14 g, 29.07 mmol) was dissolved in tetrahydrofuran (150 mL), and aqueous sodium hydroxide solution (58.2 mL, 2.5 M) was added. The mixture was stirred for 4 h. Concentrated hydrochloric acid solution was added to adjust the pH to about 3, and the resulting mixture was concentrated under reduced pressure to give crude title compound 6g (21.2 g), which was directly used in the next step without purification.

[0287] MS m/z (ESI): 454.0 [M+1].

Step 7

(6-methyl-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 6

[0288] Compound 6g (250 mg, 330.75 μmol), 4-oxa-7-azaspiro[2.5]octane hydrochloride (49.5 mg, 330.77 μmol) and HATU (233.4 mg, 992.28 μmol) were dissolved in N,N-dimethylformamide (30 mL), and N,N-diisopropylethylamine (256 mg, 1.98 mmol) was added. The mixture was stirred for 2 h. Water (60 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (80 mL×3). The organic phases were combined, washed with saturated brine (80 mL×3), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 6 (40 mg, yield: 22%).

[0289] MS m/z (ESI): 549.1 [M+1].

[0290] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 7.52-7.49 (m, 2H), 7.42-7.35 (m, 4H), 6.71-6.68 (m, 1H), 4.89 (d, 2H), 4.09 (t, 2H), 3.92 (s, 3H), 3.74-3.57 (m, 9H), 3.29 (s, 1H), 2.41 (t, 4H), 0.72-0.61 (m, 4H).

Example 7

(7-chloro-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 7

[0291] ##STR00042##

Step 1

Ethyl 2-(7-chloro-4-oxothiochroman-3-yl)-2-oxoacetate 7b

[0292] Sodium ethoxide (62.0 g, 182.2 mmol, 20% w/w ethanol solution) was added into a 500 mL single-neck flask, and a solution of diethyl oxalate (19.9 g, 136.2 mmol) in toluene (300 mL) was added at 0 TC, followed by addition of 7-chlorothiochroman-4-one 7a (18.0 g, 90.6 mmol, prepared according to the method disclosed in “Organic Letters, 2020, 22(3), 1155-1159”). The mixture was stirred at room temperature for 17 h. The reaction solution was concentrated under reduced pressure, water (400 mL) was added to the residue, and dichloromethane (200 mL×2) was added for extraction. The aqueous phase was adjusted to pH of about 2 with 5 M hydrochloric acid solution and extracted with ethyl acetate (200 mL×3). The organic phases were combined, washed with saturated brine (200 mL×2), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the title compound 7b (13.7 g), which was directly used in the next step without purification.

Step 2

Ethyl 2-(7-chloro-1,1-dioxido-4-oxothiochroman-3-yl)-2-oxoacetate 7c

[0293] Crude compound 7b (11.80 g, 36.15 mmol) was dissolved in dichloromethane (150 mL), and 3-chloroperoxybenzoic acid (17.2 g, 81.79 mmol) was added. The mixture was stirred at room temperature for 17 h. Then the mixture was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system B to give the title compound 7c (12.3 g, yield: 98.3%).

Step 3

Ethyl 7-chloro-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylate 5,5-dioxide 7d

[0294] Compound 7c (11.3 g, 33.6 mmol) was dissolved in ethanol (80 mL), and the hydrochloride of compound 2c (9.0 g) and glacial acetic acid (20 mL) were added. The mixture was stirred at 80° C. for 2 h. Then the mixture was concentrated under reduced pressure, slurried with ethanol and filtered, and the filter cake was dried in vacuum to give the title product 7d (13.9 g, yield: 83.2%).

Step 4

7-chloro-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylic acid 5,5-dioxide 7e

[0295] Compound 7d (13.9 g, 28.3 mmol) was dissolved in tetrahydrofuran (100 mL), and aqueous sodium hydroxide solution (3 M, 5.5 mL) was added. The mixture was stirred at room temperature for 16 h. The reaction solution was adjusted to pH of about 2 with 5.0 M hydrochloric acid solution and concentrated under reduced pressure to give the title product 7e (16.1 g), which was directly used in the next step without purification.

Step 5 (7-chloro-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 7

[0296] Crude compound 7e (200 mg, 255.2 μmol), 4-oxa-7-azaspiro[2.5]octane hydrochloride (49.5 mg, 330.77 μmol) and HATU (233.4 mg, 992.28 μmol) were dissolved in N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (256 mg, 1.98 mmol) was added. The mixture was stirred for 3 h. Water (60 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (80 mL×3). The organic phases were combined, washed with saturated brine (80 mL×3), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 7 (55 mg, yield: 32.7%).

[0297] MS m/z (ESI): 569.1 [M+1].

[0298] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 8.00 (s, 1H), 7.72 (d, 1H), 7.61-7.47 (m, 4H), 6.89 (d, 1H), 4.94 (d, 2H), 4.05 (s, 1H), 3.90 (s, 1H), 3.76-3.59 (m, 10H), 2.42 (brs, 4H), 0.68-0.64 (m, 4H).

Example 8

(1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-6-(trifluoromethyl)-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 8

[0299] ##STR00043##

Step 1

3-((2-(trifluoromethyl)phenyl)thio)propanoic acid 8b

[0300] Compound 8a (10.4 g, 58.3 mmol) was dissolved in N,N-dimethylformamide (60 mL), and potassium carbonate (16.1 g, 116.5 mmol) was added. The mixture was stirred at 60° C. for 30 min. Then the mixture was cooled, and bromopropionic acid (9.8 g, 64.3 mmol) was added. The resulting mixture was stirred at 60° C. for 3 h. Then the mixture was cooled, poured into water, adjusted to pH of 2 with 2 M hydrochloric acid and extracted three times with ethyl acetate. The organic phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system B to give the title compound 8b (11.0 g, yield: 75.3%).

Step 2

8-(trifluoromethyl)thiochroman-4-one 8c

[0301] Compound 8b (11.0 g, 46.5 mmol) was added to concentrated sulfuric acid (150 mL), and the mixture was stirred at room temperature for 3 h. The reaction solution was poured into ice water and stirred well. Then the reaction solution was filtered and washed with water. The solid was dissolved in ethyl acetate, washed with saturated brine, dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 8c (8.0 g, yield: 71.8%).

Step 3

Ethyl 2-(8-(trifluoromethyl)-4-oxothiochroman-3-yl)-2-oxoacetate 8d

[0302] Sodium ethoxide (23.5 g, 69.1 mmol, 20% w/w ethanol solution) was added to a 500 mL single-neck flask, and a solution of diethyl oxalate (7.6 g, 51.7 mmol) in toluene (200 mL) was added at 0° C., followed by the addition of compound 8c (8.0 g, 34.5 mmol). The mixture was stirred at room temperature for 17 h. The reaction solution was concentrated under reduced pressure, water (300 mL) was added to the residue, and dichloromethane (100 mL×2) was added for extraction. The aqueous phase was adjusted to pH of about 2 with 5 M hydrochloric acid solution and extracted with ethyl acetate (100 mL×3). The organic phases were combined, washed with saturated brine (200 mL×2), dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give crude title compound 8d (9.1 g), which was directly used in the next step without purification.

Step 4

Ethyl 2-(8-(trifluoromethyl)-1,1-dioxido-4-oxothiochroman-3-yl)-2-oxoacetate 8e

[0303] Compound 8d (9.1 g, 27.2 mmol) was dissolved in dichloromethane (150 mL), and 3-chloroperoxybenzoic acid (11.6 g, 57.2 mmol) was added. The mixture was stirred at room temperature for 17 h. Then the mixture was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system B to give the title compound 8e (9.8 g, yield: 99.3%).

Step 5

Ethyl 6-(trifluoromethyl)-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylate 5,5-dioxide 8f

[0304] Compound 8e (9.8 g, 26.9 mmol) was dissolved in ethanol (80 mL), and the hydrochloride of compound 2c (7.0 g) and glacial acetic acid (20 mL) were added. The mixture was stirred at 80° C. for 2 h. Then the mixture was concentrated under reduced pressure, slurried with ethanol and filtered, and the filter cake was dried to give the title product 8f (8.0 g, yield: 55.5%).

Step 6

6-(trifluoromethyl)-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylic acid 5,5-dioxide 8g

[0305] Compound 8f (8.0 g, 14.9 mmol) was dissolved in tetrahydrofuran (100 mL), and aqueous sodium hydroxide solution(3 M, 5.5 mL) was added. The mixture was stirred at room temperature for 16 h. The reaction solution was adjusted to pH of about 2 with 5.0 M hydrochloric acid solution and concentrated under reduced pressure to give the title product 8g (10.5 g, purity: 70%), which was directly used in the next step without purification.

Step 7

(1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-6-(trifluoromethyl)-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 8

[0306] Crude compound 8g (200 mg, 255.2 μmol), 4-oxa-7-azaspiro[2.5]octane hydrochloride (49.5 mg, 330.77 μmol) and HATU (233.4 mg, 992.28 μmol) were dissolved in N,N-dimethylformamide (10 mL), and N,N-diisopropylethylamine (172.9 mg, 1.34 mmol) was added. The mixture was stirred for 3 h. Water (60 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated brine (80 mL×2), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title compound 8 (60 mg, yield: 37.2%).

[0307] MS m/z (ESI): 603.1 [M+1].

[0308] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 8.01 (d, 1H), 7.72 (t, 1H), 7.52 (t, 2H), 7.46-7.41 (m, 2H), 7.19 (d, 1H), 5.00 (d, 2H), 4.11 (s, 1H), 3.95 (s, 1H), 3.75-3.52 (m, 10H), 2.41 (brs, 4H), 0.69-0.65 (m, 4H).

Example 9

(7-methyl-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 9

[0309] ##STR00044## ##STR00045##

Step 1

3-(m-tolylthio)propionic acid 9b

[0310] 3-methylbenzenethiol 9a (10 g, 80.5 mmol) was dissolved in N,N-dimethylformamide (100 mL), and potassium carbonate (16 g, 115.7 mmol) was added, followed by addition of 3-bromopropionic acid with stirring at room temperature. The mixture was stirred at room temperature for 2 h. Water (500 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (80 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give crude title compound 9b (8.94 g, yield: 56.5%), which was directly used in the next step without purification.

[0311] MS m/z(ESI): 195.0 [M−1].

Step 2

7-methylthiochroman-4-one 9c

[0312] Crude compound 9b (8.94 g, 45.5 mmol) was dissolved in concentrated sulfuric acid (100 mL), and the mixture was stirred at room temperature for 3 h. The reaction solution was carefully poured into ice water (500 g) to quench the reaction, and the aqueous phase was extracted with ethyl acetate (80 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography with developing solvent system B to give the title compound 9c (5.17 g, yield: 63.7%).

[0313] .sup.1H NMR (500 MHz, CDCl.sub.3) δ 7.30 (t, 1H), 7.20 (d, 1H), 7.03 (d, 1H), 3.27-3.21 (m, 2H), 2.93-2.85 (m, 2H), 2.49 (s, 3H).

Step 3

Ethyl 2-(7-methyl-4-oxothiochroman-3-yl)-2-oxoacetate 9e

[0314] Sodium ethoxide (20 g, 58.78 mmol, 20% w/w ethanol solution) was added to a single-neck flask and cooled in an ice bath. Diethyl oxalate (4.5 g, 30.79 mmol, dissolved in 50 mL of toluene) was added, followed by addition of compound 9c (5.17 g, 29.00 mmol, dissolved in 50 mL of toluene) with stirring. The mixture was stirred at room temperature for 18 h. The reaction solution was concentrated under reduced pressure, and water (200 mL) was added to quench the reaction. The pH was adjusted to neutral with 3 M aqueous hydrochloric acid solution, and the aqueous phase was extracted with ethyl acetate (100 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give crude title compound 9e (7.87 g, yield: 97.5%).

[0315] MS m/z(ESI): 279.0 [M+1].

Step 4

Ethyl 2-(7-methyl-1,1-dioxido-4-oxothiochroman-3-yl)-2-oxoacetate 9f

[0316] Compound 9e (7.78 g, 28.28 mmol) was dissolved in dichloromethane (240 mL), and 3-chloroperoxybenzoic acid (14 g, 68.96 mmol) was added in portions under an ice bath. The mixture was stirred at room temperature for 3 h. Insoluble substances were filtered off, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography with system A to give the title compound 9f (6.88 g, yield: 78.2%).

[0317] MS m/z(ESI): 310.9 [M+1].

Step 5

Ethyl 7-methyl-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylate 5,5-dioxide 9g

[0318] Compound 9f (6.88 g, 22.17 mmol) and the hydrochloride of compound 2c (4.5 g, 21.71 mmol) were dissolved in ethanol (150 mL), and glacial acetic acid (2.5 g, 41.63 mmol) was added. The mixture was stirred at reflux for 6 h. After the reaction solution was cooled to room temperature, a yellow solid was filtered off, collected and dried in vacuum to give crude title product 9g (14.7 g), which was directly used in the next step without purification.

[0319] MS m/z(ESI): 482.2 [M+1].

Step 6

7-methyl-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylic acid 5,5-dioxide 9h

[0320] Crude compound 9g (5 g, 10.38 mmol) was dissolved in tetrahydrofuran (60 mL), and aqueous sodium hydroxide solution (2.5 M, 10 mL) was added. The mixture was stirred at 60° C. for 1 h. After the reaction solution was cooled to room temperature, the pH was adjusted to neutral with 3 M hydrochloric acid. The organic solvent was removed under reduced pressure and the remaining aqueous phase was lyophilized to give crude title product 9h (9 g), which was directly used in the next step without purification.

[0321] MS m/z(ESI): 452.1 [M−1].

Step 7

(7-methyl-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 9

[0322] Crude compound 9h (200 mg, 0.22 mmol), 4-oxa-7-azaspiro[2.5]octane hydrochloride (40 mg, 0.27 mmol) and HATU (120 mg, 0.32 mmol) were dissolved in N,N-dimethylformamide (5 mL), and N,N-diisopropylethylamine (100 mg, 0.77 mmol, 0.13 mL) was added. The mixture was stirred for 2 h. Water (150 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated brine (50 mL×3), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system A to give the title product 9 (32 mg, yield: 26.5%).

[0323] MS m/z(ESI): 549.1 [M+1].

[0324] .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 7.84 (s, 1H), 7.54 (t, 2H), 7.49 (d, 1H), 7.45 (d, 1H), 7.38 (d, 1H), 6.73 (d, 1H), 4.84 (d, 2H), 4.10-4.03 (m, 1H), 3.91 (s, 1H), 3.73 (d, 2H), 3.70-3.64 (m, 2H), 3.64-3.56 (m, 6H), 2.44-2.36 (m, 7H), 0.75-0.66 (m, 2H), 0.66-0.58 (m, 2H).

Control Example 10

(6-fluoro-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(7-oxa-4-azaspiro[2.5]octan-4-yl)methanone 10

[0325] ##STR00046##

[0326] Compound 1a, 7-oxa-4-azaspiro[2.5]octane hydrochloride (33 mg, 220.77 mmol, Jiangsu Aikon), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (251 mg, 651.77 mmol), 1-hydroxybenzotriazole (99 mg, 655.77 mmol), N,N-diisopropylethylamine (110 mg, 1.09 mmol) and 4-dimethylaminopyridine (53 mg, 437.18 mmol) were dissolved in dichloromethane (30 mL), and the mixture was stirred at room temperature for 16 h. Water (50 mL) was added, and a mixed solvent (60 mL) of dichloromethane and methanol (V:V=10:1) was added for extraction. The organic phases were combined, washed successively with water (60 mL) and saturated brine (60 mL), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the resulting residue was purified by a CombiFlash rapid preparation instrument with eluent system A to give the title product 10 (25 mg, yield: 20.6%).

[0327] MS m/z (ESI): 553.3 [M+1].

[0328] .sup.1H NMR (500 MHz, DMSO-d.sub.6): δ 7.60-7.44 (m, 6H), 6.65 (d, 1H), 4.95 (d, 2H), 3.99-3.56 (m, 9H), 2.41 (s, 4H), 1.27-0.72 (m, 7H).

Example 11

(7-chloro-6-fluoro-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 11

[0329] ##STR00047##

Step 1

3-((3-chloro-2-fluorophenyl)thio)propanoic acid 11b

[0330] 3-chloro-2-fluorobenzenethiol 11a (8 g, 49.20 mmol, Wuxi Kehua) was dissolved in N,N-dimethylformamide (100 mL), and potassium carbonate (8.840 g, 63.96 mmol) was added. The mixture was stirred at 60° C. for 30 min, followed by addition of 3-bromopropionic acid (8.279 g, 54.12 mmol). The resulting mixture was stirred at 60° C. for 2 h. Water (500 mL) was added to quench the reaction, and ethyl acetate (200 mL×1) was added for extraction. The aqueous phase was adjusted to pH of about 3 with concentrated hydrochloric acid and then extracted with ethyl acetate (300 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give crude title compound 11b (11.166 g, yield: 96.7%), which was directly used in the next step without purification.

Step 2

7-chloro-8-fluorothiochroman-4-one 11c

[0331] Crude compound 11b (11.116 g, 47.37 mmol) was dissolved in concentrated sulfuric acid (100 mL), and the mixture was stirred at room temperature for 3 h. The reaction solution was carefully poured into ice water (500 mL) to quench the reaction, and the aqueous phase was extracted with ethyl acetate (200 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give the title compound 11c (8.731 g, yield: 85.1%), which was directly used in the next step without purification.

Step 3

Ethyl 2-(7-chloro-8-fluoro-4-oxothiochroman-3-yl)-2-oxoacetate 11d

[0332] Sodium ethoxide (27.423 g, 80.60 mmol, 20% w/w ethanol solution, TCI) and toluene (125 mL) were added to a single-neck flask. The mixture was cooled in an ice bath, and diethyl oxalate (8.834 g, 60.45 mmol) was added, followed by addition of crude compound 11c (8.731 g, 40.30 mmol) with stirring. The resulting mixture was stirred at room temperature for 17 h. The reaction solution was concentrated under reduced pressure, and water (600 mL) was added to quench the reaction. The pH was adjusted to about 3 with concentrated hydrochloric acid, and the aqueous phase was extracted with ethyl acetate (250 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give crude title compound 11d (11.919 g, yield: 93.4%).

[0333] MS m/z(ESI): 316.9 [M+1].

Step 4

Ethyl 2-(7-chloro-8-fluoro-1,1-dioxido-4-oxothiochroman-3-yl)-2-oxoacetate 11e

[0334] Compound 11d (11.919 g, 37.63 mmol) was dissolved in dichloromethane (130 mL), and 3-chloroperoxybenzoic acid (19.100 g, 94.08 mmol) was added in portions under an ice bath. The mixture was stirred at room temperature for 17 h. Insoluble substances were filtered off, the filtrate was concentrated, and the residue was purified by column chromatography with system A to give the title compound 11e (11.5 g, yield: 87.6%).

[0335] MS m/z(ESI): 347.0 [M−1].

Step 5

Ethyl 7-chloro-6-fluoro-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylate 5,5-dioxide 11f

[0336] Compound 11e (11.5 g, 32.98 mmol) and the hydrochloride of compound 2c (10.024 g) were dissolved in ethanol (250 mL), and glacial acetic acid (3.961 g, 65.96 mmol) was added. The mixture was stirred at reflux for 3 h. Saturated sodium bicarbonate solution (300 mL) was added to the reaction solution, and ethyl acetate (250 mL×4) was added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give crude title product 11f (16 g, yield: 93.3%), which was directly used in the next step without purification.

[0337] MS m/z(ESI): 520.0 [M+1].

Step 6

7-chloro-6-fluoro-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylic acid 5,5-dioxide 11g

[0338] Crude compound 11f (16 g, 30.77 mmol) was dissolved in tetrahydrofuran (250 mL), and aqueous sodium hydroxide solution (2.5 M, 62 mL) was added. The mixture was stirred at room temperature for 4 h. The reaction solution was adjusted to pH of about 3 with 3 M hydrochloride acid and concentrated under reduced pressure to give crude title product 11g (15 g, yield: 99.1%), which was directly used in the next step without purification.

[0339] MS m/z(ESI): 491.9 [M+1].

Step 7

(7-chloro-6-fluoro-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 11

[0340] Crude compound 11g (506 mg, 0.52 mmol), 4-oxa-7-azaspiro[2.5]octane hydrochloride (80 mg, 0.53 mmol) and HATU (240 mg, 0.63 mmol) were dissolved in N,N-dimethylformamide (25 mL), and N,N-diisopropylethylamine (340 mg, 2.63 mmol) was added. The mixture was stirred for 17 h. Saturated sodium bicarbonate solution (50 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (50 mL×3). The organic phases were combined, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography with eluent system A to give the title product 11 (72.6 mg, yield: 23.6%).

[0341] MS m/z(ESI): 587.0 [M+1].

[0342] .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 7.85 (s, 1H), 7.53-7.46 (m, 4H), 6.68-6.66 (m, 1H), 5.05-5.03 (m, 2H), 4.06-3.47 (m, 12H), 2.50-2.42 (m, 4H), 0.73-0.60 (m, 4H).

Example 12

(9-methyl-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 12

[0343] ##STR00048## ##STR00049##

Step 1

5-methylthiochroman-4-one 12a

[0344] Crude compound 9b (8.94 g, 45.5 mmol) was dissolved in concentrated sulfuric acid (100 mL), and the mixture was stirred at room temperature for 3 h. The reaction solution was carefully poured into ice water (500 g) to quench the reaction. Liquid separation was performed, and the aqueous phase was extracted with ethyl acetate (80 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography with developing solvent system B to give the title compound 12a (5.17 g, yield: 41.6%).

[0345] .sup.1H NMR (500 MHz, CDCl.sub.3) δ 7.30 (t, 1H), 7.20 (d, 1H), 7.03 (d, 1H), 3.27-3.21 (m, 2H), 2.93-2.85 (m, 2H), 2.49 (s, 3H).

Step 2

Ethyl 2-(5-methyl-4-oxothiochroman-3-yl)-2-oxoacetate 12b

[0346] Sodium ethoxide (13 g, 38.21 mmol, 20% w/w ethanol solution) was added to a single-neck flask and cooled in an ice bath. Diethyl oxalate (3.0 g, 20.53 mmol, dissolved in 50 mL of toluene) was added, followed by addition of compound 12a (3.38 g, 18.93 mmol, dissolved in 50 mL of toluene) with stirring. The mixture was stirred at room temperature for 18 h. The reaction solution was concentrated under reduced pressure, and water (200 mL) was added to quench the reaction. The pH was adjusted to neutral with 3 M aqueous hydrochloric acid solution, and the aqueous phase was extracted with ethyl acetate (100 mL×3). The organic phases were combined, dried over anhydrous sodium sulfate and filtered, and the filtrate was concentrated under reduced pressure to give crude title compound 12b (3.66 g, yield: 69.56%), which was directly used in the next step without purification.

[0347] MS m/z(ESI): 279.0 [M+1].

Step 3

Ethyl 2-(5-methyl-1,1-dioxido-4-oxothiochroman-3-yl)-2-oxoacetate 12c

[0348] Compound 12b (3.66 g, 13.17 mmol) was dissolved in dichloromethane (50 mL), and 3-chloroperoxybenzoic acid (6 g, 29.55 mmol, content: 85%) was added in portions under an ice bath. The mixture was stirred at room temperature for 3 h. Insoluble substances were filtered off, the filtrate was concentrated under reduced pressure, and the residue was purified by column chromatography with system A to give crude title compound 12c (7.3 g, crude product).

[0349] MS m/z(ESI): 310.9 [M+1].

Step 4

Ethyl 9-methyl-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylate 5,5-dioxide 12d

[0350] Crude compound 12c (7.3 g, 12.94 mmol) and the hydrochloride of compound 2c (3.6 g) were dissolved in ethanol (50 mL), and glacial acetic acid (1.5 g, 24.98 mmol) was added. The mixture was stirred at reflux for 6 h. After being cooled to room temperature, the reaction solution was filtered, and the filter cake was collected and dried in vacuum to give the title product 12d (6.8 g), which was directly used in the next step without purification.

[0351] MS m/z(ESI): 482.2 [M+1].

Step 5

9-methyl-1-(4-(morpholinomethyl)phenyl)-1,4-dihydrothiochromeno[4,3-c]pyrazole-3-carboxylic acid 5,5-dioxide 12e

[0352] Crude compound 12d (1.35 g, 1.54 mmol) was dissolved in tetrahydrofuran (15 mL), and aqueous sodium hydroxide solution (2.5 M, 3 mL) was added. The mixture was stirred at 60° C. for 1 h. After the reaction solution was cooled to room temperature, the pH was adjusted to neutral with 3 M hydrochloric acid. The organic solvent was removed under reduced pressure and the remaining aqueous phase was lyophilized to give crude title product 12e (2.2 g), which was directly used in the next step without purification.

[0353] MS m/z(ESI): 452.1 [M−1].

Step 6

(9-methyl-1-(4-(morpholinomethyl)phenyl)-5,5-dioxido-1,4-dihydrothiochromeno[4,3-c]pyrazol-3-yl)(4-oxa-7-azaspiro[2.5]octan-7-yl)methanone 12

[0354] Crude compound 12e (200 mg, 0.22 mmol), 4-oxa-7-azaspiro[2.5]octane hydrochloride (45 mg, 0.30 mmol) and HATU (150 mg, 0.40 mmol) were dissolved in N,N-dimethylformamide (5 mL), and N,N-diisopropylethylamine (150 mg, 1.16 mmol, 0.19 mL) was added. The mixture was stirred for 2 h. Water (150 mL) was added to quench the reaction, and the aqueous phase was extracted with ethyl acetate (50 mL×3). The organic phases were combined, washed with saturated brine (50 mL×3), dried over anhydrous sodium sulfate and filtered, the filtrate was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography with eluent system A to give the title product 12 (62 mg, yield: 43.7%).

[0355] MS m/z(ESI): 549.1 [M+H].

[0356] .sup.1H NMR (500 MHz, DMSO-d.sub.6) δ 7.89 (d, 1H), 7.64 (t, 1H), 7.53 (d, 1H), 7.45 (t, 2H), 7.27 (d, 2H), 4.79 (s, 2H), 4.06 (t, 1H), 3.93 (s, 1H), 3.79-3.47 (m, 9H), 3.29 (s, 1H), 2.37 (s, 4H), 1.61 (d, 3H), 0.75-0.52 (m, 4H).

TEST EXAMPLES

Biological Evaluation

Test Example 1. Test on Inhibitory Activity and Selectivity of Compounds Disclosed Herein on PI3Kδ Enzyme

[0357] I. Experimental Objective

[0358] This experiment is intended to test the inhibitory effect and selectivity of the compounds on PI3Kδ enzyme activity, and to evaluate the in vitro activity of the compounds according to the IC.sub.50 values.

[0359] II. Experimental Principle

[0360] In this experiment, an ADP-Glo™ kinase assay kit is used. A substrate is phosphorylated under the action of enzyme, and ADP is generated at the same time. An ADP-Glo Reagent is added to remove unreacted ATP in the reaction system, and the ADP generated by the reaction is detected by a kinase detection reagent. In the presence of a compound, the inhibition rate of the compound is calculated by measuring the signal value.

[0361] III. Experimental Materials

[0362] 1. Instruments

TABLE-US-00002 Instrument Supplier Model Centrifuge Eppendorf 5430 Microplate reader Perkin Elmer Envision, SN. 1050214 Echo 550 Labcyte Echo 550

[0363] 2. Reagents and Consumables

TABLE-US-00003 Reagent Supplier Cat. No. PIK3CD/PIK3R1 Carna 11-103 PI103 selleckchem S1038 DMSO Sigma D8418-1L 384-well white plate PerkinElmer 6007290

[0364] IV. Experimental Procedures

[0365] Test compounds were each subjected to 3-fold dilution from a starting concentration of 10000 nM to obtain 11 concentrations, and duplicate wells were set for the test. Gradient dilution to a corresponding 100-fold final concentration in a 384-well plate was performed to obtain solutions with 11 different concentrations. 50 nL of each solution was transferred to compound wells of a 384-well plate with Echo; negative control wells were added with 50 nL of DMSO. A kinase solution with a 2-fold final concentration was prepared using 1×kinase buffer. 2.5 μL of the kinase solution with a 2-fold final concentration was added to the compound wells; 2.5 μL of the 1× kinase buffer was added to the negative control wells. The mixture was centrifuged at 1000 rpm for 30 s, mixed well by shaking, and then incubated at room temperature for 10 min. A mixed solution of ATP and substrate P1P2 with a 2-fold final concentration was prepared using 1× kinase buffer. The reaction was initiated by adding 2.5 μL of the mixed solution of ATP and substrate with a 2-fold final concentration. The mixture in the 384-well plate was centrifuged at 1000 rpm for 30 s, mixed well by shaking, and then reacted at room temperature for 120 min. 5 μL of ADP-Glo reagent was added, and the mixture was centrifuged at 1000 rpm for 30 s, mixed well by shaking, and then incubated at room temperature for 40 min. 10 μL of kinase detection reagent was added, and the resulting mixture was centrifuged at 1000 rpm for 30 s, mixed well by shaking, and then incubated at room temperature for 30 min. The luminescence RLU was read with an Envision microplate reader.

[0366] V. Data Analysis

[0367] IC.sub.50 values for the inhibitory activity of the compounds were calculated using Graphpad Prism software. The results are shown in Table 1 below.

TABLE-US-00004 TABLE 1 Data on inhibition and selectivity activity of compounds disclosed herein on PI3Kδ enzyme (nM) PI3Kδ PI3Kα PI3Kβ PI3Kγ enzyme enzyme enzyme enzyme Example No. IC.sub.50 IC.sub.50 IC.sub.50 IC.sub.50 1 13 467.6 1316.0 6210.0 2 18.4 — — — 3 5.6 — — — 4 7.9 — — — 5 19 2329 2791 >10000 6 68 2570 7058 >10000 7 26.5 1523 1743 >10000 9 25.0 1761 1551 >10000 11 10.5 441 736 8918 12 5.8 — — — 10 (Control >10000 — — — Example)

CONCLUSION

[0368] compared with the Control Example, the compounds disclosed herein have stronger inhibitory activity and selectivity on PI3Kδ enzyme.