CN103626787B

CN103626787B - Thienothio compound and application thereof

Info

Publication number: CN103626787B
Application number: CN201310666934.4A
Authority: CN
Inventors: 胡春; 王欣; 金辄; 刘晓平; 黄二芳
Original assignee: Shenyang Pharmaceutical University
Current assignee: Shenyang Pharmaceutical University
Priority date: 2013-12-10
Filing date: 2013-12-10
Publication date: 2016-06-15
Anticipated expiration: 2033-12-10
Also published as: CN103626787A

Abstract

The invention belongs to pharmaceutical technology field, relate to thienothiapyran compound and application thereof. Thienothiapyran compound includes the derivant of thienothiapyran compound and pharmaceutically useful salt, its general structure is as follows: the salt of thienothiapyran compound and the pharmaceutically useful acid addition of this compounds can merge with existing medicine or alone as somatomedin tyrosine kinase inhibitor on epidermis, the relevant disease such as small cell lung cancer of imbalance is forwarded to for treating EGF-R ELISA signal, scale cancer, adenocarcinoma, large cell carcinoma, colorectal carcinoma, breast carcinoma, ovarian cancer, renal cell carcinoma, bronchial asthma.

Description

Thienothio compound and application thereof

Technical field

The invention belongs to pharmaceutical technology field, relate to Thienothio compound and apply as epidermal growth factor recipient tyrosine kinase inhibitor, and preparation method thereof.

Background technology

Differentiation degree according to cancerous cell and morphological characteristic, pulmonary carcinoma can be divided into nonsmall-cell lung cancer and small cell lung cancer. Research finds, there is the imbalance of substantial amounts of epidermal growth factor signal transduction and the overexpression of epidermal growth factor recipient tyrosine kinase in patients with lung cancer.

Known EGF-R ELISA (epidermalgrowthfactorreceptor, EGFR) as a member of receptor type tyrosine kinase family, being a kind of transmembrane glycoprotein, it is made up of extracellular epidermal growth factor land (comprising 621 amino acid residues), hydrophobic transmembrane domain (23 amino acid residues), intracellular kinases district (542 amino acid residues) and carboxyl terminal four part. EGFR has 4 kinds of type HER-1, HER-2, HER-3 and HER-4, when smaller ligand is combined with EGFR, EGFR is made to activate, and then the tyrosine kinase district of EGFR activates, identify the substrates enzymes of albumen, in incoming for signal cell, also will can activating many downstream signal transduction paths after EGFR activation simultaneously, stimulating cellular growth is bred. Owing to receptor type tyrosine kinase Main Differences is the outer ligand binding domains of born of the same parents, and the tyrosine kinase domain in born of the same parents has higher homology, it is contemplated that the smaller ligand that the outer ligand binding domain of synthesis born of the same parents is well combined, thus suppressing Bao Nei tyrosine kinase activity district, the catalysis activity of inhibitory enzyme and tyrosine autophosphorylation, and then suppress the transfer etc. of cell cycle progression, angiogenesis and tumor.

Existing epidermal growth factor recipient tyrosine kinase inhibitor, such as gefitinib, Erlotinib, Lapatinib etc., all also exists the dermoreactions such as diarrhoea, erythra, pruritus, and possible headache, heart QT intervals extend and bioavailability reduction etc.

Compound of the present invention, as the epidermal growth factor recipient tyrosine kinase inhibitor of brand new type, has structure type novelty, the obvious feature of drug action. Can be used for the relevant disease such as small cell lung cancer treated or prevent and the imbalance of EGF-R ELISA signal transduction causes, scale cancer, adenocarcinoma, large cell carcinoma, colorectal carcinoma, breast carcinoma, ovarian cancer, renal cell carcinoma, bronchial asthma, there is good using value and development prospect.

Summary of the invention

Technical problem solved by the invention is to provide a kind of compound as shown in Formulas I and II, its prodrug and pharmaceutical active metabolite and its pharmaceutically acceptable salt, and provides its application in the medicine of the preparation prevention disease relevant with treatment EGFR signal transduction imbalance.

Wherein

N is 1,2;

R₁Can be H or halogen;

R₂It can be hydrogen or halogen;

R₃For by 1,2 or 3 independently selected from H, halogen, C₁-C₄Alkyl, C₁-C₄Alkoxyl ,-CF₃、-OCF₃The phenyl ring replaced, benzhydryl, 4-chlorobenzhydryl, benzyl;

R₄、R₅Separately selected from hydrogen, C₁-C₄Alkyl, N, N-diethyl, phenyl, furfuryl, benzyl, it is preferable that hydrogen, methyl, ethyl, the tert-butyl group, isopropyl, N, N-diethyl, phenyl, furfuryl, or R₂、R₃Pyrrolidinyl, piperidyl, N-methyl piperidine base, morpholinyl, hexamethylene imine basic ring is formed together with the nitrogen-atoms being connected with them.

" pharmaceutically acceptable salt " refers to remain Formulas I, the biopotency of II compound and character, and the conventional acid addition salts formed with suitable non-toxic organic or inorganic acid or organic or inorganic alkali or base addition salts. the example of acid-addition salts includes acetate, adipate, alginate, aspartate, benzoate, benzene sulfonate, disulfate, butyrate, citrate, Camphora hydrochlorate, camsilate, cipionate, digluconate, lauryl sulfate, esilate, fumarate, gluceptate, glycerophosphate, Hemisulphate, enanthate, caproate, hydrogen chlorate, hydrobromate, hydriodate, 2-isethionate, lactate, maleate, mesylate, 2-naphthalene sulfonate, nicotinate, nitrate, oxalates, pamoate, pectinic acid salt, persulfate, 3-phenylpropionic acid salt, picrate, Pivalate, propionate, succinate, sulfate, tartrate, rhodanate, toluene fulfonate and undecylate. alkali salt includes ammonium salt, alkali metal salt, for instance sodium and potassium salt, alkali salt, for instance calcium and magnesium salt, the salt of organic base, such as hexanamine salt, N-methyl-D-glucamine salt, and amino acid whose salt, such as arginine, lysine etc., and, Basic nitrogen-containing groups can be quaternized with such reagent, for instance elementary alkyl halide, such as methyl, the chlorine of ethyl, propyl group and butyl, bromine and iodide, dialkyl sulfate, such as dimethyl sulfate, diethylester, dibutyl ester and diamyl ester, long chain halide, such as the chlorine of decyl, lauryl, myristyl and stearyl, bromine and iodide, aralkyl halide, such as the bromide etc. of benzyl and phenethyl. the acid being preferred for generating acid-addition salts includes hydrochloric acid and acetic acid.

" pharmaceutically acceptable " such as pharmaceutically acceptable ground carrier, excipient, prodrug etc., refers to pharmacologically acceptable and to administration particular compound the essentially no toxicity of patient.

" pharmaceutical active metabolite " refers to the metabolite of pharmaceutically acceptable and effective Formulas I, II compound.

The present invention also relates to suppress the Pharmaceutical composition of epidermal growth factor recipient tyrosine kinase, said composition contains Formulas I, II compound or derivant or its pharmaceutically useful acid-addition salts and pharmaceutically useful carrier.

In the present invention, the term " halogen " of application includes chlorine, bromine or fluorine.

" replacement ", unless otherwise indicated, refers to that substituent group can exist in one or more positions, and substituent group is independently selected from specifically option.

The compounds of this invention can be taken to patient by diverse ways, for instance is administered orally with capsule or tablet, is administered with sterile solution agent or suspensoid, and in some cases, it is possible to solution form intravenous injection. Can carry out preparing and taking with its pharmaceutically useful acid addition salt form thereof by the free alkali compound of the present invention.

Detailed description of the invention

Reaction process 1 summarises the synthesis step preparing the compounds of this invention.

Reaction process 1

The present invention is described in detail with following example. However, it should be understood that the invention is not restricted to the following example of concrete narration.

The preparation of embodiment 1:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-aminomethyl phenyl) piperazine (compound number 01)

The preparation of step A:5,6-dihydro-4H-thieno [2,3-b] thiapyran-4-ketone

100mL round-bottomed flask adds 4.64g (0.04mol) 2-mercapto-thiophene, 80mL oxolane, the lower dropping 11mL triethylamine of stirring, add 3.3mL acrylic acid, be heated to reflux 12h. React complete, boil off oxolane. After cooling, add 40mL ethyl acetate and 20mL water, adjust pH to 2, collected organic layer with 18% hydrochloric acid. Aqueous layer with ethyl acetate extracts, and merges organic layer, dries with anhydrous magnesium sulfate. Sucking filtration, filtrate boils off solvent, precipitates out brown solid after cooling. Petroleum ether recrystallization obtains white needles, yield 6.02g, yield 80.0%, fusing point 43-45 DEG C.

100mL round-bottomed flask adds 3.76g (0.02mol) 3-(thiophene-2-sulfenyl) propanoic acid, 20mL dichloromethane and 2 DMF; under nitrogen protection; dropping dichloromethane solution (the oxalyl chloride 2.2mL containing oxalyl chloride while stirring; dichloromethane 16mL), 1h is stirred at room temperature. The bath of reactant liquor cryosel is cooled to-10 DEG C, while stirring the dropping dichloromethane solution (butter of tin 1.15mL, dichloromethane 10mL) containing butter of tin. Drip and finish, at 0 DEG C, stir 0.5h. In flask, add 20mL water, extract organic facies, aqueous phase dichloromethane extraction, merge organic facies. Organic facies with saturated solution of sodium carbonate, water and sodium chloride saturated solution washing, uses anhydrous magnesium sulfate successively. Sucking filtration, filtrate boils off solvent, obtains crude product, yield 100% after cooling. Petroleum ether recrystallization obtains white needles, yield 2.59g, yield 76.2%, fusing point 59-60 DEG C.

The preparation of step B:4,5,6,7-Tetramethylene sulfide also [2,3-b] thia-4-ketone

In the round-bottomed flask of 500mL, add little sodium block (6.6g, 0.29mol), under dry room temperature, add dehydrated alcohol (120mL). Cool down after sodium is wholly absent, add 2-mercapto-thiophene, under room temperature condition, stir about 30min, the dimmed redness of reactant liquor. Addition-butyrolactone (15.6mL, 0.3mol), reacts about 19h reactant liquor and becomes viscous under heated reflux condition. Product at reduced pressure is distilled, removes ethanol. Remaining solid is washed by about 100mL petroleum ether, ethyl acetate (V:V=3:1).Being added by solid after washing in the 1N hydrochloric acid of cooling, bottom precipitates out dark red oil, with saturated NaCl solution washing organic layer after being extracted with ethyl acetate, dries with anhydrous MgSO4. Elimination desiccant, concentration. Concentrated solution adds petroleum ether and is about 250mL, stop stirring and heating after being heated to reflux about 1h, stand. Pouring in conical flask after the clarification of question response liquid, solution becomes muddiness, is transferred in another conical flask after standing clarification. Freeze overnight precipitates out white crystal. Yield 41.26%.

The dichloromethane of 4-(thiophene-2-sulfenyl) butanoic acid (4.04g, 0.02mol) and 25ml is added in 250ml three-necked bottle. Drip the dichloromethane solution (25mL) of oxalyl chloride (5.4mL, 0.06mol), the thin out yellow green of reactant liquor when nitrogen protection, drip stirring 2h. Under nitrogen protection and condition of ice bath, being slowly added dropwise the dichloromethane solution (20ml) of butter of tin (2.3mL, 0.02mol), reactant liquor becomes blackish green. 24h is stirred under room temperature condition. In reactant liquor, add 50ml hydrochloric acid (V:V=1:1), stir to not having gas to release. Washing organic layer, becomes orange-yellow to pH weakly acidic pH organic layer. With saturated NaCl solution washing organic layer, separation MgSO₄Dry. White crystal is obtained with petroleum ether recrystallization. Yield 27%.

The preparation of the bromo-1,4-dihydro-thiophene of step C:7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-formic acid

100mL round-bottomed flask adds 0.46g metallic sodium and 20mL absolute methanol, is heated to reflux to sodium solids disappeared, boils off methanol. In reaction bulb add 2.36g(0.02mol) dimethyl oxalate., 20mL toluene, 1.70g (0.01mol) 5,6-dihydro-4H-thieno [2,3-b] thiapyran-4-ketone. 24h is stirred at room temperature, reactant liquor is poured in 100mL frozen water, extract aqueous phase. Organic facies 20mL10% sodium hydroxide solution extracts. Merging aqueous phase, after washing with absolute ether, adjust pH to 2 with 18% dilute hydrochloric acid, obtain yellow solid, sucking filtration obtains crude product. Petroleum ether recrystallization obtains yellow needle-like crystals 1.5g, yield 43.86%, fusing point: 93-95 DEG C.

100mL round-bottomed flask adds 2-oxo-2-(4-oxo-5,6-dihydro-4H-thieno [2,3-b] thiapyran-5-base) methyl acetate 0.51g (2mmol) and 10mL glacial acetic acid, stirring is lower adds hydrazine hydrate 2mL(3.2mmol), it is heated to reflux 12h. Being poured into by reactant liquor in 200mL frozen water and stir, white solid occur, sucking filtration obtains crude product. Petroleum ether recrystallization obtains white solid 0.32g, yield 63.2%, fusing point: 94-96 DEG C.

100mL round-bottomed flask adds 1,4-dihydro-thiophene also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carboxylate methyl ester 2.52g(0.01mol), 50mL glacial acetic acid, 5mL water, bromine 2.08g(0.013mol is dripped under ice bath cooling and stirring), room temperature reaction 24h after completion of dropwise addition. Reactant liquor is poured in 200mL frozen water, precipitate out white solid, sucking filtration, obtain product. ESI-MS:m/z330.9 [M]⁺。

100mL round-bottomed flask adds 0.62g(2mmol) the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carboxylate methyl ester, 20mL water and 0.4g sodium hydroxide, it is heated to reflux 2h. Reactant liquor adjusts pH to 2 with the hydrochloric acid of 18%, white solid occurs, and sucking filtration obtains crude product. Petroleum ether recrystallization obtains white solid 0.57g, yield 90.0%.

The preparation of step D:4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carboxylic acid

Sodium (0.23g, 0.01mol) being cut into sodium bits, adds in 10ml absolute methanol, stir to sodium solids disappeared, rotation steaming boils off methanol, obtains white solid.In reaction bulb, add (COOMe) 2 (1.18g, 0.01mol), the toluene of 4,5,6,7-Tetramethylene sulfides also [2,3-b] thia-5-ketone (1g, 0.005mol) and 7ml, 40h is stirred at room temperature. Pouring in the frozen water of about 100ml, layer of fetching water, water layer is kermesinus. With the NaOH aqueous solution extraction organic layer of 10%, combining water layer. Aqueous washed with ether. The hcl acidifying of 1N, in acidization, liquid becomes muddiness, obtains yellow turbid liquid and put into standing in refrigerator when being adjusted to pH=2. Precipitate out solid very thickness, use CH₂Cl₂Sticky solid is obtained after extracting concentration. Red powder, yield 40.29% is obtained with absolute ether washing.

(4-oxo-4,5,6,7-Tetramethylene sulfides are [2,3-b] thia-5-base also) acetonic acid methyl ester (0.5g, 2mmol) and hydrazine hydrate (0.1g, 2mmol) are added in round-bottomed flask, adds 5mL glacial acetic acid, be heated to reflux 3h. Reactant liquor is poured in frozen water, precipitate out yellow solid, filter, dry. Yield 79%.

100mL round-bottomed flask adds 0.51g (2mmol) 4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carboxylate methyl ester, 20mL water and 0.4g sodium hydroxide, is heated to reflux 2h. Reactant liquor adjusts pH to 2 with the hydrochloric acid of 18%, white solid occurs, and sucking filtration obtains crude product. Petroleum ether recrystallization obtains white solid 0.31g, yield 65.1%. 1H-NMR (300MHz, DMSO): δ 2.74 (t, 2H), 3.26 (m, 2H), 7.27 (d, 1H, J=5.4Hz), 7.63 (d, 1H, J=5.3Hz). (13.13 s, 1H)

The preparation of the bromo-1,4-dihydro-thiophene of step E:7,8-bis-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carboxylic acid

3.31g(0.01mol) bromo-Isosorbide-5-Nitrae-the dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carboxylate methyl ester adds in acetic acid (50mL)-water (5mL) mixed solution, drips bromine (8g under room temperature condition, 0.05mol), 48h is stirred at room temperature. Reactant liquor is poured into (100g) in frozen water mixed liquor, precipitate out light yellow solid. Sucking filtration, column chromatography chromatogram separation purification (petroleum ether: ethyl acetate=10:1), obtain white solid, yield 8%. ESI-MS:m/z410.7 [M+H]⁺

7,8-bis-bromo-Isosorbide-5-Nitrae-dihydro-thiophenes also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carboxylate methyl ester (0.82g, 2mmol) is joined in 30mL water, adds sodium hydroxide (0.1g, 2.5mmol), be heated to reflux until solids disappeared. Reactant liquor is cooled to room temperature, 18% salt acid for adjusting pH value, to no longer having solid to precipitate out, sucking filtration, obtain white solid, yield 90%.

The preparation of step F:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-aminomethyl phenyl) piperazine (compound number 01)

By bromo-for 7-Isosorbide-5-Nitrae-dihydro-thiophene also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-formic acid (1.27g, 4mmol), 4-aminomethyl phenyl piperazine (0.74g, 4.2mmol), 1-(3-dimethylamino-propyl)-3-ethyl-carbodiimide hydrochloride (1g, 5mmol), I-hydroxybenzotriazole (0.1g, 0.74mmol), triethylamine (2mL) adds in 20mL dichloromethane, 48h being stirred at room temperature, filters, filtrate is respectively with 18% hydrochloric acid, saturated sodium carbonate and water washing. Extracting organic facies, anhydrous magnesium sulfate dries. Filtering, filtrate boils off solvent, separates (petroleum ether: ethyl acetate=5:1) through silica gel column chromatography, obtains white solid. Yield 30%. M.p.:185-186 DEG C;IR (KBr, cm^-1):3441,2920,2852,1610,1513,1445,1384,1156,1023,813,619;¹H-NMR(600MHz,CDCl₃): δ 7.24 (1H, s), 6.91 (2H, dd, J=7.2Hz, 2.4Hz), 6.85 (2H, dd, J=7.2Hz, 2.4Hz), 4.28 (2H, s), 3.99 (7H, m), 3.13 (4H, m); ESI-MS (m/z): 345.1 [M+H]⁺。

The preparation of embodiment 2:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(diphenyl methyl) piperazine (compound number 02)

According to the method for example 1, prepare 1-(the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-is [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl also)-4-(diphenyl methyl) piperazine 0.77g, yield 35%. M.p.:175-176 DEG C; IR (KBr, cm^-1):3420,2921,2851,1607,1451,1384,1262,1154,1027,997,877,831,705;¹H-NMR(600MHz,CDCl₃): δ 2.44 (4H, m), 3.79 (4H, m), 4.16 (2H, s), 4.25 (1H, s), 7.19 (2H, d, J=8.4Hz), 7.28 (4H, dd, J=7.2Hz, 8.4Hz), 7.41 (4H, d, J=7.2Hz); ESI-MS (m/z): 551.2 [M+H]⁺。

The preparation of the bromo-1,4-dihydro-thiophene of embodiment 3:N, N-diethyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide (compound number 03)

According to the method for example 1, prepare the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of N, N-diethyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide (compound number 03) 0.75g, yield 50%. M.p.:147-148 DEG C; IR (KBr, cm^-1):3212,2958,2917,2849,1735,1583,1535,1508,1485,1461,1375,1215,1159,967,849;¹H-NMR(600MHz,CDCl₃): δ 1.25 (6H, m), 3.54 (4H, m), 4.12 (2H, s), 7.72 (1H, s); ESI-MS (m/z): 372.0,374.0 [M+H]⁺。

The preparation of embodiment 4:1-(7-bromo-1,4-dihydro-thiophene also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(the chloro-4-fluorophenyl of 3-) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(the chloro-4-fluorophenyl of 3-) piperazine (compound number 04) 0.72g, yield 36%. M.p.:120 DEG C; IR (KBr, cm^-1):3233,2924,2853,1745,1609,1586,1502,1464,1382,1223,1154,998;¹H-NMR(600MHz,CDCl₃): δ 3.16 (4H, t, J=6.0Hz), 3.95 (4H, t, J=6.0Hz), 4.26 (2H, s), 6.79 (1H, m), 6.95 (1H, dd, J=6.3Hz, J=2.4Hz), 7.05 (1H, dd, J=8.7Hz, J=9Hz), 7.25 (1H, s); ESI-MS (m/z): 512.8,514.9,517.0 [M+H]⁺。

The preparation of embodiment 5:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-chlorphenyl) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-chlorphenyl) piperazine (compound number 05) 0.50g, yield 25%. M.p.:235 DEG C; IR (KBr, cm^-1):3159.9,2918.2,2849.7,1740.4,1590.8,1496.6,1480.7,1441.4,1387.7,1234.2,120.6,1151.0,998.1,810.2;¹H-NMR(600MHz,DMSO-d₆): δ 3.10 (4H, m), 3.51 (4H, m), 4.26 (2H, s), 7.21 (1H, s), 7.56 (2H, d, J=5.4Hz), 7.58 (2H, d, J=5.4Hz), 13.29 (1H, s); ESI-MS (m/z): 497.3,495.4,499.2 [M+H]⁺。

The preparation of embodiment 6:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-Trifluoromethoxyphen-l) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-Trifluoromethoxyphen-l) piperazine (compound number 06) 0.92g, yield 42%. M.p.:228-230 DEG C; IR (KBr, cm^-1):3240,2919,2850,1730,1605,1532,1510,1475,1386,1368,1327,1272,1153,1028,1004;¹H-NMR(600MHz,DMSO-d₆): δ 3.33 (8H, m), 4.27 (2H, s), 7.03 (2H, d, J=8.6Hz), 7,20 (2H, d, J=8.6Hz), 7.56 (1H, s), 13.84 (1H, s); ESI-MS (m/z): 547.0,545.1 [M+H]⁺。

The preparation of embodiment 7:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(2-fluorophenyl) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(2-fluorophenyl) piperazine (compound number 07) 0.75g, yield 39%.M.p.:78-80 DEG C; IR (KBr, cm^-1):3221,2921,2851,1720,1641,1500,1446,1394,1385,1287,1240,1151,999;¹H-NMR(600MHz,CDCl₃): δ 3.11 (4H, t, J=10.2Hz), 4.02 (4H, t, J=10.2Hz), 4.24 (2H, s), 7.02 (3H, m), 7.23 (1H, s), 7.38 (1H, d, J=7.8Hz); ESI-MS (m/z): 479.1,481.0 [M+H]⁺。

The preparation of embodiment 8:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-trimethoxyphenyl) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-trimethoxyphenyl) piperazine (compound number 08) 0.87g, yield 44%. M.p.:90 DEG C; IR (KBr, cm^-1):2922,1730,1618,1511,1446,1379,1255,1224,1186,1137,1096,1034,992,816;¹H-NMR(600MHz,CDCl₃): δ 3.13 (4H, m), 3.99 (7H, m), 4.28 (2H, s), 6.85 (2H, dd, J=7.2Hz, 2.4Hz), 6.91 (2H, dd, J=7.2Hz, 2.4Hz), 7.24 (1H, s); ESI-MS (m/z): 491.1,493.2 [M+H]⁺。

The preparation of embodiment 9:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-phenylpiperazine

According to the method for example 1, prepare 1-(the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-is [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl also)-4-phenylpiperazine (compound number 09) 0.85g, yield 43%. M.p.:112 DEG C; IR (KBr, cm^-1):2923,2853,1619,1517,1499,1468,1377,1276,1154,992,832;¹H-NMR(600MHz,CDCl₃): δ 3.26 (4H, t), 4.01 (4H, t), 4.27 (2H, s), 6.91 (1H, d, J=7.8Hz), 6.96 (2H, d, J=7.8Hz), 7,28 (2H, dd, J=7.8Hz, 7.8Hz), 7.25 (1H, s); ESI-MS (m/z): 461.1,463.1 [M+H]⁺。

The preparation of embodiment 10:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-fluorophenyl) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-fluorophenyl) piperazine (compound number 10) 0.67g, yield 35%. M.p.:185-186 DEG C; IR (KBr, cm^-1):3443,2922,2852,1605,1507,1443,1384,1225,1116;¹H-NMR(600MHz,CDCl₃): δ 3.19 (4H, t), 4.04 (4H, t), 4.24 (2H, s), 7.00 (4H, m), 7.24 (1H, s); ESI-MS (m/z): 479.1,481.0 [M+H]⁺。

The preparation of embodiment 11:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-methyl piperazine

According to the method for example 1, prepare 1-(the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-is [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl also)-4-methyl piperazine (compound number 11) 0.95g, yield 60%. M.p.:220-222 DEG C; IR (KBr, cm^-1):3424.0,2917.8,849.5,1704.1,1465.5,1433.1,1383.9,1296.8,1204.9,940.2;¹H-NMR(600MHz,CDCl₃): δ 2.36 (3H, s), 2.51 (4H, t), 3.86 (4H, t), 4.19 (2H, s), 7.28 (1H, s); ESI-MS (m/z): 399.2,401.2 [M+H]⁺。

The preparation of embodiment 12:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-benzyl diethylenediamine

According to the method for example 1, prepare 1-(the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-is [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl also)-4-benzyl diethylenediamine (compound number 12) 0.89g, yield 47%. M.p.:180 DEG C; IR (KBr, cm^-1):3422,3206,2918,2850,1730,1601,1509,1446,1384,1266,1126,1029,999;¹H-NMR(600MHz,CDCl₃): δ 2.50 (4H, t), 3.54 (2H, s), 3.77 (4H, t), 4.28 (2H, s), 7.24 (1H, s), 7.31 (5H, m); ESI-MS (m/z): 474.9,477.1 [M+H]⁺。

The preparation of embodiment 13:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-[(4-chlorphenyl) (phenyl) methyl] piperazine

Method according to example 1, prepare 1-(7-bromo-1,4-dihydro-thiophene also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-[(4-chlorphenyl) (phenyl) methyl] piperazine (compound number 13) 0.89g, yield 38%.M.p.:212-214 DEG C; IR (KBr, cm^-1):3442,2920,2851,1602,1452,1384,1126,998;¹H-NMR(600MHz,CDCl₃): δ 2.44 (4H, m), 3.79 (4H, m), 4.16 (2H, s), 4.25 (1H, s), 7.22 (2H, m), 7,25 (1H, s), 7.27 (1H, m), 7.29 (2H, dd, J=7.8Hz, 7.8Hz), 7.37 (4H, m); ESI-MS (m/z): 584.9,586.8 [M+H]⁺。

The preparation of the bromo-1,4-dihydro-thiophene of the embodiment 14:N-tert-butyl group-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide

According to the method for example 1, prepare the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of the N-tert-butyl group-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide (compound number 14) 1.12g, yield 75%. M.p.:205-207 DEG C; IR (KBr, cm^-1):3364,3201,2919,1638,1553,1526,1492,1450,1385,1296,1220,1167,969,863;¹H-NMR(600MHz,CDCl₃): δ 1.49 (9H, s), 4.49 (2H, s), 7,21 (1H, s), 7.40 (1H, s); ESI-MS (m/z): 372.0,374.2 [M+H]⁺。

The preparation of embodiment 15:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(3-bromophenyl) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(3-bromophenyl) piperazine (compound number 15) 0.82g, yield 38%. M.p.:150-151 DEG C; IR (KBr, cm^-1):3418.7,3217.5,2920.0,2850.7,1590.4,1480.6,1442.9,1384.6,1224.6,1157.9,999.9,938.2,764.3;¹H-NMR(600MHz,CDCl₃): δ 3.25 (4H, m), 4.00 (4H, m), 4.26 (2H, s), 6.85 (1H, m), 7.01 (1H, m), 7.06 (1H, m), 7.13 (1H, m), 7.23 (1H, s); ESI-MS (m/z): 538.9,540.8,542.9 [M+H]⁺。

The preparation of embodiment 16:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-ethoxyl phenenyl) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-ethoxyl phenenyl) piperazine (compound number 16) 0.89g, yield 44%. M.p.:178-180 DEG C; IR (KBr, cm^-1):3443.4,2918.4,2850.2,1618.7,1511.5,1446.0,1384.5,1273.2,1254.0,1224.2,1137.0,1021.6,993.6,834.2,816.7;¹H-NMR(600MHz,CDCl₃): δ 1.40 (3H, t), 3.13 (4H, t), 3.99 (6H, m), 4.28 (2H, s), 6.85 (2H, dd, J=7.2Hz, 2.4Hz), 6.91 (2H, dd, J=7.2Hz, 2.4Hz), 7.24 (1H, s); ESI-MS (m/z): 505.3,507.3 [M+H]⁺。

The preparation of embodiment 17:4-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl) morpholine

According to the method for example 1, prepare 4-(the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-is [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl also) morpholine (compound number 17) 0.76g, yield 49%. M.p.:150 DEG C; IR (KBr, cm^-1):3444.9,3169.7,2918.2,2850.3,1729.9,1592.2,1441.1,1266.5,1148.2,1113.9,1000.1,969.6,844.0;¹H-NMR(600MHz,DMSO-d₆): δ 3.61 (4H, t), 3.96 (4H, t), 4.28 (2H, s), 7.42 (1H, s), 13.59 (1H, s); ESI-MS (m/z): 388.2 [M+H]⁺。

The preparation of embodiment 18:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl) pyrrolidine

According to the method for example 1, prepare 1-(the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-is [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl also) pyrrolidine (compound number 18) 0.77g, yield 52%. M.p.:220 DEG C; IR (KBr, cm^-1):3442.7,2920.1,2851.2,1606.7,1575.4,1508.2,1454.8,1384.3,1156.9,969.2,844.5;¹H-NMR(600MHz,DMSO-d₆): δ 1.81 (2H, t), 1.87 (2H, t), 3.46 (2H, t), 3.84 (2H, t), 4.37 (2H, s), 7.42 (1H, s), 14.57 (1H, s); ESI-MS (m/z): 370.1,372.3 [M+H]⁺。

The preparation of the bromo-1,4-dihydro-thiophene of embodiment 19:N-isopropyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide

According to the method for example 1, prepare the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of N-isopropyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide (compound number 19) 0.99g, yield 69%.M.p.:189-190 DEG C; IR (KBr, cm^-1):3400.5,3207.5,2974.5,2921.3,1637.4,1556.3,1528.3,1452.1,1384.6,1175.4,967.5,836.5;¹H-NMR(600MHz,DMSO-d₆): δ 4.50 (1H, m), 4.39 (2H, s), 7.50 (1H, s), 8.08 (1H, s); ESI-MS (m/z): 357.1,359.1 [M+H]⁺。

The preparation of embodiment 20:N-(2-diethylin) ethyl-7-bromo-1,4-dihydro-thiophene also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide

According to the method for example 1, prepare the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of N-(2-diethylin) ethyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide (compound number 20) 1.06g, yield 64%. M.p.:165-167 DEG C; IR (KBr, cm^-1):3420.4,2968.1,2921.8,1635.0,1556.8,1528.9,1456.6,1384.3,1124.4,968.5,831.9;¹H-NMR(600MHz,CDCl₃): δ 1.07 (6H, t), 2.68 (4H, m), 2.77 (2H, t), 3.55 (2H, t), 4.43 (2H, s), 7.21 (1H, s), 8.05 (1H, s); ESI-MS (m/z): 415.3 [M+H]⁺。

The preparation of the bromo-1,4-dihydro-thiophene of embodiment 21:N, N-dimethyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide

According to the method for example 1, prepare the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of N, N-dimethyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide (compound number 21) 0.94g, yield 68%. M.p.:220 DEG C; IR (KBr, cm^-1):3443.5,3205.2,2920.4,1612.5,1536.4,1485.5,1420.6,1384.3,1259.4,1160.8,1101.3,966.7,822.5;¹H-NMR(600MHz,DMSO-d₆): δ 2.97 (6H, s), 4.26 (2H, s), 7.43 (1H, s), 13.54 (1H, s); ESI-MS (m/z): 344.2,346.3 [M+H]⁺。

The preparation of the bromo-1,4-dihydro-thiophene of embodiment 22:N-phenyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide

According to the method for example 1, prepare the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of N-phenyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide (compound number 22) 0.47g, yield 30%. M.p.:190-191 DEG C; IR (KBr, cm^-1):3443.8,2919.8,2850.5,1730.5,1612.4,1555.1,1441.0,1384.2,1296.2,1114.0,1038.4,778.5;¹H-NMR(600MHz,DMSO-d₆): δ 4.42 (2H, s), 7.40 (5H, m), 7.42 (1H, s), 8.82 (1H, s), 13.64 (1H, s); ESI-MS (m/z): 415.3,417.3 [M+Na]⁺。

The preparation of embodiment 23:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl) piperidines

According to the method for example 1, prepare 1-(the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-is [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl also) piperidines (compound number 23) 0.95g, yield 62%. M.p.:196-198 DEG C; IR (KBr, cm^-1):3442.6,2931.0,2852.9,1603.1,1570.1,1538.8,1445.3,1384.3,274.9,1112.4,966.6,851.8;¹H-NMR(600MHz,CDCl₃): δ 1.66 (6H, m), 3.72 (4H, t), 4.14 (2H, s), 7.30 (1H, s); ESI-MS (m/z): 384.2,386.3 [M+H]⁺,406.2,408.4[M+Na]⁺。

The preparation of the bromo-1,4-dihydro-thiophene of embodiment 24:N benzyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide

According to the method for example 1, prepare the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of N benzyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide (compound number 24) 0.91g, yield 56%. M.p.:98-100 DEG C; IR (KBr, cm^-1):3421.9,2920.9,2851.7,1629.7,1446.5,1384.2,1274.3,1114.1,850.9;¹H-NMR(600MHz,DMSO-d₆): δ 4.40 (2H, d), 4.42 (2H, s), 7.40 (5H, m), 7.42 (1H, s), 8.82 (1H, s), 13.64 (1H, s); ESI-MS (m/z): 408.3 [M+H]⁺,430.1[M+Na]⁺。

The preparation of embodiment 25:N (furan-2-methylene) the bromo-1,4-dihydro-thiophene of-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide

According to the method for example 1, prepare N (furan-2-methylene) bromo-Isosorbide-5-Nitrae-dihydro-thiophene of-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide (compound number 25) 0.62g, yield 39%. M.p.:89-90 DEG C; IR (KBr, cm^-1):3444.2,2921.3,2851.3,16307,1440.4,1384.0,1236.2,1112.3,1089.9,975.6,737.9;¹H-NMR(600MHz,CDCl₃): δ 4.22 (2H, s), 4.60 (2H, d), 6.29 (1H, d, J=6Hz), 6.33 (1H, d, J=6Hz), 7.20 (1H, dd, J=6Hz, 6Hz), 7.25 (1H, s);ESI-MS (m/z): 395.3,397.2 [M+H]⁺,418.3,420.1[M+Na]⁺。

The preparation of embodiment 26:N (pyridine-2-base) the bromo-1,4-dihydro-thiophene of-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide

According to the method for example 1, prepare N (pyridine-2-base) bromo-Isosorbide-5-Nitrae-dihydro-thiophene of-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide (compound number 26) 0.49g, yield 31%. M.p.:83-84 DEG C; IR (KBr, cm^-1):3421.0,2921.4,2851.5,1630.3,1445.7,1384.2,1274.8,1126.2;¹H-NMR(600MHz,CDCl₃): δ 4.31 (2H, s) .6.50 (1H, d, J=8.2Hz), 6.65 (1H, m), 7.42 (2H, m), 8.07 (1H, d, J=4.8Hz); ESI-MS (m/z): 395.2 [M+H]⁺。

The preparation of embodiment 27:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(3,5-bis-trifluoromethyl) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(3,5-bis-trifluoromethyl) piperazine (compound number 27) 1.08g, yield 45%. M.p.:220-221 DEG C; IR (KBr, cm^-1):3421.2,2918.2,1610.2,1581.2,1485.5,1400.4,1275.0,1175.,1131.4,995.3,963.6,862.4;¹H-NMR(600MHz,DMSO-d₆): δ 3.45 (4H, t), 3.77 (2H, t), 4.14 (2H, t), 4.31 (2H, s), 7.33 (1H, s), 7.45 (1H, s), 7.50 (2H, s), 13.64 (1H, s); ESI-MS (m/z): 596.9,599.1 [M+H]⁺,619.1,621.0[M+Na]⁺。

The preparation of embodiment 28:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(3,4-difluorophenyl) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(3,4-difluorophenyl) piperazine (compound number 28) 0.88g, yield 44%. M.p.:190-192 DEG C; IR (KBr, cm^-1):3444.2,2919.8,1656.7,1560.4,1417.7,1384.3,1276.6,1116.2;¹H-NMR(600MHz,DMSO-d₆): δ 3.15 (4H, t), 3.72 (4H, t), 4.27 (2H, s), 6.75 (1H, m), 7.03 (1H, m), 7.25 (1H, d, J=9.6Hz), 7.45 (1H, s); ESI-MS (m/z): 518.9,521.0 [M+Na]⁺。

The preparation of embodiment 29:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(2-aminomethyl phenyl) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(2-aminomethyl phenyl) piperazine (compound number 29) 0.91g, yield 48%. M.p.:167-170 DEG C; IR (KBr, cm^-1):3442.6,2920.4,2851.6,1621.6,1491.9,1443.3,1384.0,1327.3,1274.9,1224.2,1126.5,1021.8,1000.2,762.3;¹H-NMR(600MHz,CDCl₃): δ 2.35 (3H, s), 2.98 (4H, t), 3.97 (4H, t), 4.23 (2H, s), 7.01 (2H, m), 7.20 (2H, m), 7.25 (1H, s); ESI-MS (m/z): 475.1,476.4 [M+H]⁺。

The preparation of embodiment 30:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(2-chlorphenyl) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(2-chlorphenyl) piperazine (compound number 30) 0.73g, yield 37%. M.p.:185-186 DEG C; IR (KBr, cm^-1):3444.2,2920.3,2851.4,1610.1,1478.0,1384.4,1265.9,1227.6,1124.4,1022.3;¹H-NMR(600MHz,CDCl₃): δ 3.11 (4H, t), 4.02 (4H, t), 4.24 (2H, s), 7.02 (3H, m), 7.23 (1H, s), 7.38 (1H, d, J=7.8Hz); ESI-MS (m/z): 495.1,497.1 [M+H]⁺。

The preparation of embodiment 31:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(2,5-3,5-dimethylphenyl) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(2,5-3,5-dimethylphenyl) piperazine (compound number 31) 0.92g, yield 47%.M.p.:150-152 DEG C; IR (KBr, cm^-1):3443.5,2919.6,2850.9,1606.7,1457.4,1384.1,1125.3;¹H-NMR(600MHz,CDCl₃): δ 2.29 (3H, s), 2.30 (3H, s), 2.95 (4H, t), 3.96 (4H, t), 4.27 (2H, s), 6.82 (1H, s), 6.84 (1H, d, J=7.2Hz), 7.08 (1H, d, J=7.2Hz), 7.25 (1H, s); ESI-MS (m/z): 489.1,491.1 [M+H]⁺。

The preparation of embodiment 32:1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(3-trifluoromethyl) piperazine

According to the method for example 1, prepare the 1-(bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(3-trifluoromethyl) piperazine (compound number 32) 0.95g, yield 45%. M.p.:185-186 DEG C; IR (KBr, cm^-1):3444.9,3219.1,2918.2,2849.8,1591.1,1444.4,1384.4,1349.9,1319.6,1269.0,1222.1,1163.5,1112.2,1075.2,999.6,946.4,864.1,785.5;¹H-NMR(600MHz,DMSO-d₆): δ 3.31 (8H, t), 4.27 (2H, s), 7.09 (d, J=7.7Hz, 1H), 7.24 (d, J=8.4Hz, 1H), 7.43 (t, J=8.0Hz, 1H), 7.50 (1H, s); MS (m/z): 427.2,429.0 [M-H]⁺。

The preparation of embodiment 33:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(benzhydryl phenyl) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(benzhydryl phenyl) piperazine (compound number 33) 0.74g, yield 38%. M.p.:138-139 DEG C; IR (KBr, cm^-1):3422.1,3207.1,2919.3,2851.1,1617.2,1491.3,1448.8,1384.3,1285.9,1242.7,1142.9,995.3,859.8,747.1,706.2;¹H-NMR(600MHz,CDCl₃): δ 2.44 (4H, t), 3.08 (2H, t), 3.47 (2H, t), 3.76 (4H, t), 5.25 (1H, s), 7.19 (2H, d, J=7.2Hz), 7.22 (1H, d, J=5.4Hz), 7.26 (1H, d, J=5.4Hz), 7.28 (4H, t, J=7.2Hz), 7.41 (4H, d, J=7.2Hz); MS (m/z): 487.2 [M+H]⁺。

The preparation of embodiment 34:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-chlorphenyl) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-chlorphenyl) piperazine (compound number 34) 0.60g, yield 35%. M.p.:140-141 DEG C; IR (KBr, cm^-1):3381.8,2920.9,2850.4,1613.8,1493.2,1440.8,1383.9,1342.3,1270.7,1230.9,1155.5,1021.5,998.6,883.5,860.0,825.0;¹H-NMR(600MHz,CDCl₃): δ 3.10 (2H, t), 3.21 (4H, t), 3.47 (2H, t), 3.94 (4H, t), 6.90 (2H, d, J=9.0Hz), 7.12 (2H, d, J=9.0Hz), 7.23 (1H, d, J=5.4Hz), 7.26 (1H, d, J=5.4Hz); MS (m/z): 431.1 [M+H]⁺。

The preparation of embodiment 35:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-[(4-chlorphenyl) (phenyl) methyl] piperazine

Method according to example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-[(4-chlorphenyl) (phenyl) methyl] piperazine (compound number 35) 0.87g, yield 42%. M.p.:142-143 DEG C; IR (KBr, cm^-1):3432.2,2919.3,2851.4,1618.6,1488.1,1445.5,1384.4,1286.8,1241.9,1142.8,995.7,860.1,757.4,719.2;¹H-NMR(600MHz,CDCl₃): δ 2.32 (4H, t), 3.02 (2H, t), 3.38 (2H, t), 3.68 (4H, t), 4.16 (1H, s), 7.12 (d, J=5.4Hz, 1H), 7.19 (d, J=5.4Hz, 1H), 7.24 (3H, m), 7.28 (2H, d, J=5.4Hz), 7.33 (4H, m); MS (m/z): 521.2 [M+H]⁺。

The preparation of the embodiment 36:N-tert-butyl group-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide

According to the method for example 1, prepare the N-tert-butyl group-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide (compound number 36) 0.64g, yield 52%. M.p.:170-171 DEG C;IR (KBr, cm^-1):3393.2,3253.5,2962.0,2921.2,2851.0,1644.8,1521.4,1484.0,1384.6,1363.0,1258.8,1217.8,1124.6,1053.3,877.9,865.2;¹H-NMR(600MHz,CDCl₃): δ 1.47 (9H, s), 3.10 (2H, t), 3.69 (2H, t), 7.21 (1H, d, J=5.4Hz), 7.24 (1H, d, J=5.4Hz); MS (m/z): 308.2 [M+H]⁺。

The preparation of embodiment 37:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-methyl piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-methyl piperazine (compound number 37) 0.60g, yield 45%. M.p.:107-108 DEG C; IR (KBr, cm^-1):3429.0,3228.4,2915.6,2792.8,1608.8,1504.8,1439.5,1352.5,1290.1,1253.7,1225.3,1142.0,997.9,861.7,766.6;¹H-NMR(600MHz,CDCl₃): δ 2.33 (3H, s), 2.46 (4H, t), 3.10 (2H, t), 3.46 (2H, t), 3.91 (4H, t), 7.23 (1H, d, J=5.4Hz), 7.26 (1H, d, J=5.4Hz); MS (m/z): 335.2 [M+H]⁺。

The preparation of embodiment 38:N, N-diethyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide

According to the method for example 1, prepare N, N-diethyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide (compound number 38) 0.69g, yield 56%. M.p.:183-185 DEG C; IR (KBr, cm^-1):3443.6,3200.5,2929.8,1601.5,1509.2,1479.3,1374.1,1284.1,1262.2,1195.5,1127.7,1092.8,1073.0,1015.8,881.7,859.0;¹H-NMR(600MHz,CDCl₃): δ 1.24 (6H, t), 3.10 (2H, t), 3.42 (2H, t), 3.54 (4H, m), 7.22 (1H, d, J=5.4Hz), 7.25 (1H, d, J=5.4Hz); MS (m/z): 308.2 [M+H]⁺。

The preparation of embodiment 39:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-aminomethyl phenyl) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-aminomethyl phenyl) piperazine (compound number 39) 0.71g, yield 43%. M.p.:186-187 DEG C; IR (KBr, cm^-1):3124.2,2916.4,1631.5,1579.3,1513.5,1443.3,1384.4,1340.9,1264.6,1236.5,1203.3,1154.1,1016.6,999.2,876.2,811.3;¹H-NMR(600MHz,CDCl₃): δ 2.28 (3H, s), 3.10 (2H, t), 3.18 (4H, t), 3.47 (2H, t), 3.93 (4H, t), 6.85 (2H, d, J=8.4Hz), 7.10 (2H, d, J=8.4Hz), 7.23 (1H, d, J=5.4Hz), 7.27 (1H, d, J=5.4Hz); MS (m/z): 411.2 [M+H]⁺。

The preparation of embodiment 40:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-phenylpiperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-phenylpiperazine (compound number 40) 0.75g, yield 47%. M.p.:74-75 DEG C; IR (KBr, cm^-1):3441.7,3209.2,2920.5,2851.6,1599.6,1495.6,1444.2,1384.1,1260.4,1230.6,1153.3,1016.0,998.3,875.4,759.5;¹H-NMR(600MHz,CDCl₃): δ 3.10 (2H, t), 3.20 (1H, t), 3.27 (4H, t), 3.47 (1H, t), 4.02 (4H, t), 6.90 (1H, m), 6.95 (2H, m), 7.23 (1H, d, J=5.4Hz), 7.28 (1H, d, J=5.4Hz), 7.29 (2H, m); MS (m/z): 397.2 [M+H]⁺。

The preparation of embodiment 41:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-Trifluoromethoxyphen-l) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-Trifluoromethoxyphen-l) piperazine (compound number 41) 0.85g, yield 44%. M.p.:85-86 DEG C; IR (KBr, cm^-1):3441.7,3209.2,2920.5,2851.6,1599.6,1495.6,1444.2,1384.1,1260.4,1230.6,1153.3,1016.0,998.3,875.4,759.5;¹H-NMR(600MHz,CDCl₃): δ 3.10 (2H, t), 3.21 (4H, t), 3.47 (2H, t), 3.94 (4H, t), 6.90 (2H, d, J=9.0Hz), 7.12 (2H, d, J=9.0Hz), 7.23 (1H, d, J=5.4Hz), 7.26 (1H, d, J=5.4Hz); MS (m/z): 481.1 [M+H]⁺。

The preparation of embodiment 42:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-Trifluoromethoxyphen-l) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-Trifluoromethoxyphen-l) piperazine (compound number 42) 0.75g, yield 45%.M.p.:195-197 DEG C; IR (KBr, cm^-1):3441.9,2919.7,2851.3,1603.7,1509.0,1444.9,1384.5,1341.8,1263.2,1229.1,1157.7,1017.4,999.5,877.7,817.2;¹H-NMR(600MHz,CDCl₃): δ 3.11 (2H, t), 3.30 (4H, t), 3.50 (2H, t), 3.98 (4H, t), 6.91 (2H, dd, J=9.1Hz, 4.2Hz), 6.99 (2H, dd, J=9.1Hz, 4.2Hz), 7.24 (1H, d, J=5.4Hz), 7.25 (1H, d, J=5.4Hz); MS (m/z): 415.1 [M+H]⁺。

The preparation of embodiment 43:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-benzyl diethylenediamine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-benzyl diethylenediamine (compound number 43) 0.80g, yield 49%. M.p.:72-73 DEG C; IR (KBr, cm^-1):3427.6,3210.5,2918.4,2851.1,2809.1,1613.9,1446.0,1384.2,1347.7,1287.2,1229.1,1143.5,997.1,860.2,740.7,699.1;¹H-NMR(600MHz,CDCl₃): δ 2.50 (4H, t), 3.10 (2H, t), 3.44 (2H, t), 3.54 (2H, s), 3.77 (4H, t), 7.24 (d, J=4.8Hz, 1H), 7.26 (d, J=4.8Hz, 1H), 7.31 (5H, m); MS (m/z): 411.2 [M+H]⁺,433.1[M+Na]⁺。

The preparation of embodiment 44:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-methoxyphenyl) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-methoxyphenyl) piperazine (compound number 44) 0.84g, yield 49%. M.p.:156-158 DEG C; IR (KBr, cm^-1):3443.8,3196.6,2920.6,2851.1,1611.7,1510.5,1445.2,1384.2,1279.1,1246.5,1225.8,1154.6,1034.6,995.2,859.4;¹H-NMR(600MHz,CDCl₃): δ 3.11 (10H, m), 3.47 (2H, t), 3.78 (3H, s), 6.85 (2H, J=9Hz), 6.92 (2H, d, J=9Hz), 7.24 (d, J=4.8Hz, 1H), 7.32 (d, J=4.8Hz, 1H); MS (m/z): 427.2 [M+H]⁺,449.2[M+Na]⁺。

The preparation of embodiment 45:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(3-trifluoromethyl) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(3-trifluoromethyl) piperazine (compound number 45) 0.80g, yield 43%. M.p.:85-87 DEG C; IR (KBr, cm^-1):3444.3,3212.2,2919.3,2851.0,1610.1,1495.4,1445.9,1384.5,1348.8,1232.6,1162.9,1120.8,995.3,860.3;¹H-NMR(600MHz,CDCl₃): δ 3.11 (2H, t), 3.30 (4H, t), 3.50 (2H, t), 3.98 (4H, t), 7.08 (1H, d, J=8.4Hz), 7.13 (2H, m), 7.25 (2H), 7.38 (1H, dd, J=8.4Hz, 8.4Hz), 10.26 (1H, s); MS (m/z): 465.1 [M+H]⁺。

The preparation of embodiment 46:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(3-bromophenyl) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(3-bromophenyl) piperazine (compound number 46) 0.76g, yield 40%. M.p.:195-197 DEG C; IR (KBr, cm^-1):3443.9,2920.9,2851.1,1589.7,1444.0,1384.1,1236.2,1124.2,994.4,860.0;¹H-NMR(600MHz,CDCl₃): δ 3.06 (2H, t), 3.24 (8H, t), 3.78 (2H, t), 6.94 (2H, m), 7.10 (1H, d), 7.15 (1H, t), 7.54 (d, J=5.4Hz, 1H), 7.56 (d, J=5.4Hz, 1H), 13.15 (1H, s); MS (m/z): 474.4,476.3 [M+H]⁺。

The preparation of embodiment 47:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(3,4-difluorophenyl) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(3,4-difluorophenyl) piperazine (compound number 47) 0.80g, yield 46%. M.p.:230-232 DEG C; IR (KBr, cm^-1):3369.6,3258.9,2920.5,2850.8,1662.4,1629.5,1555.5,1515.3,1434.9,1384.1,1280.9,1222.0,1189.0,1116.2,1004.4,870.1,805.1;¹H-NMR(600MHz,CDCl₃): δ 2.95 (4H, t), 3.15 (2H, t), 3.74 (2H, t), 3.91 (4H, t), 7.13 (1H), 7.22 (1H, d, J=5.4Hz), 7.24 (1H, m), 7.28 (1H, d, J=5.4Hz), 7.80 (1H, m);MS (m/z): 433.2 [M+H]⁺。

The preparation of embodiment 48:N, N-dimethyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide

According to the method for example 1, prepare N, N-dimethyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide (compound number 48) 0.59g, yield 53%. M.p.:232-233 DEG C; IR (KBr, cm^-1):3199.3,3153.2,3055.3,2924.6,1616.9,1524.1,1425.8,1385.1,1144.7,1088.7,1007.5,914.6,861.8,769.0,696.6;¹H-NMR(600MHz,CDCl₃): δ 3.10 (2H, t), 3.16 (6H, m), 3.45 (2H, t), 7.24 (1H, d, J=5.5Hz), 7.27 (1H, d, J=5.5Hz); MS (m/z): 280.0 [M+H]⁺。

The preparation of embodiment 49:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl) pyrrolidine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl) pyrrolidine (compound number 49) 0.60g, yield 49%. M.p.:170 DEG C; IR (KBr, cm^-1):3263.2,2920.4,2875.7,1595.8,1526.9,1492.2,1470.4,1432.4,1382.8,1339.4,1224.1,1007.6,898.7,861.2,760.5,688.5;¹H-NMR(600MHz,CDCl₃): δ 1.82 (2H, m), 1.93 (2H, m), 3.11 (2H, t), 3.37 (4H, t), 3.59 (2H, t), 7.23 (1H, d, J=5.5Hz), 7.25 (1H, d, J=5.5Hz); MS (m/z): 306.2 [M+H]⁺,328.1[M+Na]⁺。

The preparation of embodiment 50:N-isopropyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide

According to the method for example 1, prepare N-isopropyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide (compound number 50) 0.66g, yield 56%. M.p.:214-215 DEG C; IR (KBr, cm^-1):3408.8,3256.4,2922.4,1644.1,1543.5,1523.7,1485.7,1446.4,1384.0,1125.5,1007.9,859.5,760.8,692.2;¹H-NMR(600MHz,CDCl₃): δ 1.26 (6H, d), 3.12 (2H, t), 3.71 (2H, t), 4.25 (1H, m), 7.20 (1H, d, J=5.5Hz), 7.24 (1H, d, J=5.5Hz); MS (m/z): 294.1 [M+H]⁺。

The preparation of embodiment 51:N-benzyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide

According to the method for example 1, prepare N-benzyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide (compound number 51) 0.63g, yield 46%. M.p.:136-138 DEG C; IR (KBr, cm^-1):33401.9,3217.8,2921.8,2851.3,1648.5,1546.9,1452.8,1384.2,1241.6,1128.0,861.8,752.2,698.0,615.7;¹H-NMR(600MHz,CDCl₃): δ 3.12 (2H, t), 3.73 (2H, t), 4.62 (2H, d), 7.20 (1H, m), 7.23 (1H, d, J=5.5Hz), 7.28 (1H, d, J=5.5Hz), 7.34 (4H, m); MS (m/z): 342.1 [M+H]⁺。

The preparation of embodiment 52:N-phenyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide

According to the method for example 1, prepare N-phenyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide (compound number 52) 0.50g, yield 38%. M.p.:240 DEG C; IR (KBr, cm^-1):3422.8,2921.0,2850.9,1663.2,1596.1,1542.8,1440.4,1384.1,1121.3,861.5,753.4,694.0,618.0;¹H-NMR(600MHz,CDCl₃): δ 3.15 (2H, t), 3.76 (2H, t), 7.13 (1H, m), 7.22 (1H, d, J=5.5Hz), 7.28 (1H, d, J=5.5Hz), 7.37 (2H, dd, J=7.9Hz, 7.9Hz), 7.68 (2H, d, J=7.9Hz); MS (m/z): 328.1 [M+H]⁺。

The preparation of embodiment 53:4-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl) morpholine

According to the method for example 1, prepare 4-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl) morpholine (compound number 53) 0.67g, yield 52%. M.p.:176-178 DEG C; IR (KBr, cm^-1):3439.5,2920.5,2851.6,1611.7,1434.2,1271.5,1242.1,1194.6,1115.5,1024.6,995.5,861.4,619.7;¹H-NMR(600MHz,CDCl₃): δ 3.11 (2H, t), 3.48 (2H, t), 3.75 (4H, t), 3.81 (4H, t), 7.22 (1H, d, J=5.4Hz), 7.25 (1H, d, J=5.4Hz); MS (m/z): 322.0 [M+H]⁺。

The preparation of embodiment 54:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl) piperidines

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl) piperidines (compound number 54) 0.55g, yield 43%.M.p.:76-78 DEG C; IR (KBr, cm^-1):3435.9,3229.1,2920.1,2851.2,1607.8,1514.6,1445.3,1384.1,1254.9,1138.8,1027.8,989.4,859.0,774.5,617.9;¹H-NMR(600MHz,CDCl₃): δ 1.27 (2H, m), 1.64 (4H, m), 3.11 (2H, t), 3.42 (2H, t), 3.66 (4H, t), 7.23 (1H, d, J=5.4Hz), 7.24 (1H, d, J=5.4Hz); MS (m/z): 320.1 [M+H]⁺。

The preparation of embodiment 55:N-(furan-2-methylene)-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide

According to the method for example 1, prepare N-(furan-2-methylene)-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide (compound number 55) 0.46g, yield 35%. M.p.:155-156 DEG C; IR (KBr, cm^-1):3408.3,3227.4,2921.6,2850.7,1650.4,1542.9,1523.7,1484.0,1417.8,1384.2,1190.0,1147.0,1003.1,964.7,861.9,747.9,693.0;¹H-NMR(600MHz,CDCl₃): δ 3.12 (2H, t), 3.71 (2H, t), 4.60 (2H, d), 6.29 (1H, m), 6.33 (1H, m), 7.20 (1H, d, J=5.4Hz), 7.25 (1H, d, J=5.4Hz), 7.37 (1H, m); MS (m/z): 331.6 [M+H]⁺。

The preparation of embodiment 56:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(the chloro-4-fluorophenyl of 3-) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(the chloro-4-fluorophenyl of 3-) piperazine (compound number 56) 0.88g, yield 49%. M.p.:205-206 DEG C; IR (KBr, cm^-1):33434.2,3288.2,2919.2,2850.7,1603.4,1503.1,1462.9,1444.8,1384.2,1223.5,1156.2,994.4,945.2,865.3,733.5,688.5;¹H-NMR(600MHz,CDCl₃): δ 3.12 (2H, t), 3.16 (4H, t), 3.49 (2H, t), 3.95 (4H, t), 6.79 (1H, m), 6.95 (1H, dd, J=6.3Hz, 2.4Hz), 7.05 (1H, dd, J=9.0Hz, 8.7Hz), 7.23 (1H, d, J=5.4Hz), 7.25 (1H, d, J=5.4Hz); MS (m/z): 449.0,450.8 [M+H]⁺。

The preparation of embodiment 57:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(2-aminomethyl phenyl) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(2-aminomethyl phenyl) piperazine (compound number 57) 0.72g, yield 44%. M.p.:192-194 DEG C; IR (KBr, cm^-1):3442.6,3264.4,2919.3,2851.2,1601.8,1492.0,1474.7,1442.8,1383.7,1223.1,1152.0,1026.2,995.5,861.7,761.1,691.0;¹H-NMR(600MHz,CDCl₃): δ 2.33 (3H, s), 2.95 (4H, t), 3.12 (2H, t), 3.48 (2H, t), 3.91 (4H, t), 7.02 (2H, m), 7.18 (2H, m), 7.23 (1H, d, J=5.5Hz), 7.27 (1H, d, J=5.5Hz); MS (m/z): 409.0 [M-H]⁺。

The preparation of embodiment 58:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(3,5-bis-trifluoromethyl) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(3,5-bis-trifluoromethyl) piperazine (compound number 58) 0.92g, yield 43%. M.p.:89-91 DEG C; IR (KBr, cm^-1):3444.6,2920.4,1618.5,1476.8,1384.9,1277.4,1240.0,1177.9,1130.4,995.0,963.9,861.1,720.5,682.3;¹H-NMR(600MHz,CDCl₃): δ 3.12 (2H, t), 3.38 (4H, t), 3.51 (2H, t), 4.01 (4H, t), 7.23 (2H, m), 7.27 (1H, s), 7.24 (1H, s), 7.34 (1H, s); MS (m/z): 533.4 [M+H]⁺。

The preparation of embodiment 59:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(2-fluorophenyl) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(2-fluorophenyl) piperazine (compound number 59) 0.66g, yield 40%. M.p.:79-81 DEG C; IR (KBr, cm^-1):3439.4,2918.7,1611.8,1500.0,1440.3,1384.7,1342.1,1286.6,1236.6,1201.2,1147.9,998.6,884.5,860.5,747.8;¹H-NMR(600MHz,CDCl₃): δ 2.23 (2H, t), 3.12 (4H, t), 3.48 (4H, t), 3.66 (2H, t), 7.01 (1H, m), 7.03 (1H, m), 7.25 (2H, m), 7.38 (1H, m), 7.39 (1H, m);MS (m/z): 415.3 [M+H]⁺,437.1[M+Na]⁺。

The preparation of embodiment 60:1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(2-chlorphenyl) piperazine

According to the method for example 1, prepare 1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(2-chlorphenyl) piperazine (compound number 60) 0.66g, yield 38%. M.p.:203-205 DEG C; IR (KBr, cm^-1):3441.5,3214.1,2920.0,2851.1,1601.9,1500.3,1477.4,1443.6,1384.1,1336.5,1286.9,1237.1,1202.8,1146.5,1025.4,997.1,861.8,759.2,689.2;¹H-NMR(600MHz,CDCl₃): δ 2.23 (2H, t), 3.12 (4H, t), 3.48 (4H, t), 3.66 (2H, t), 7.02 (2H, m), 7.25 (2H, m), 7.38 (2H, m); MS (m/z): 431.0,432.9 [M+H]⁺。

The preparation of the bromo-1,4-dihydro-thiophene of the embodiment 61:N tert-butyl group-7,8-two also [3', 2':5,6] thiapyran also [4,3-c] pyrazoles-3-amide

According to the method for example 1, prepare the N tert-butyl group-7, the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 8-bis-also [3', 2':5,6] thiapyran also [4,3-c] pyrazoles-3-amide (compound number 61) 0.58g, yield 32%. M.p.:224-226 DEG C; IR (KBr, cm^-1):3391.7,3075.6,2935.1,1653.5,1544.7,1520.1,1439.2,1366.5,1265.9,1217.1,1018.8,987.3,966.9,858.4,822.7;¹H-NMR(600MHz,CDCl₃): δ 1.49 (9H, s), 4.49 (2H, s), 7.40 (1H, s); ESI-MS (m/z): 450.2,452.2,454.2 [M+H]⁺。

The preparation of the bromo-1,4-dihydro-thiophene of embodiment 62:N, N dimethyl-7,8-two also [3', 2':5,6] thiapyran also [4,3-c] pyrazoles-3-amide

According to the method for example 1, prepare N, N dimethyl-7, the bromo-Isosorbide-5-Nitrae-dihydro-thiophene of 8-bis-also [3', 2':5,6] thiapyran also [4,3-c] pyrazoles-3-amide (compound number 62) 0.76g, yield 42%. M.p.:159-160 DEG C; IR (KBr, cm^-1):3422.3,2977.2,2933.9,2603.8,1602.2,1476.5,1397.8,1383.3,1271.0,1172.0,1036.7,850.8,807.0;¹H-NMR(600MHz,CDCl₃): δ 1.25 (6H, m), 3.54 (4H, m), 4.12 (2H, s); ESI-MS (m/z): 450.2,452.2,454.2 [M+H]⁺。

The above, be only presently preferred embodiments of the present invention, is not the restriction that the present invention makees other form, and any those skilled in the art are changed or be modified as the Equivalent embodiments of equivalent variations possibly also with the technology contents of the disclosure above. But every without departing from technical solution of the present invention content, according to any simple modification, equivalent variations and remodeling that above example is made by the technical spirit of the present invention, still fall within the protection domain of technical solution of the present invention.

Pharmacological Examples

Embodiment 63:

The quick plate of estrogen receptor alpha measures

The combination of this mensuration detection of radioactive labels estrogen and estrogen receptor. This mensuration carries out on BioMek2000 (Beckman). Plate is at the upper reading of scintillation counter (PackardTopcount), and counting reduces explanation compound and is combined with described receptor. Described mensuration according to Allan etc., Anal.Biochem (1999), 275 (2), the method that 243-247 introduces carries out.

First day, in each hole of the 96 hole FlashPlatePlus plates cross-linked with goat anti-mouse antibody (NENLifeSciences), add 100 μ L contain 5mM dithiothreitol, DTT (DTT, Panvera), 0.5 μ g mice irritation element acceptor monoclonal antibody (SRA-1010, Stressgen) and 50ng Purification of Human estrogen receptor alpha (Panvera) estrogen screening buffer (ESB, Panvera). Seal this plate and in 4 DEG C of incubated overnight.

Second day, each hole is at room temperature washed three times with 200 μ LPBSpH7.2, and (0.5nM, is equivalent to 6nCi for every crowd of 120Ci/mmol then to add 98 μ L radioactive label estrogen in each hole, Amersham), dilute with ESB and 5mM dithiothreitol, DTT (DTT). Then in each hole, add the test compound of 2.5 μ L, 30% (v/v) dimethyl sulfoxide/50mMHEPESpH7.5 dilution.By inhaling to beat, each hole is mixed three times, seal this plate and be incubated at room temperature 1 hour. Then, in Topcount scintillation counter (Packard), each hole is counted 1 minute.

Embodiment 64:

Erss fluorescence polarization determination

The combination of this mensuration detection estrogen fluorescent analog (FluormoneES2, Panvera) and estrogen receptor. Plate can set that the upper reading of the luminescent counter for polarization mode (PackardTopCount). Relative to Vehicle controls, fluorescence reduces explanation compound and is combined with described receptor.

It is essential, in whole process, it is to avoid bubble is introduced in the reaction in the 96 each holes of orifice plate. (the bubble destruction luminous flux of reaction surface, thus polarisation-affecting reading). But, be sufficiently mixed after reactive component is joined each hole also it is critical that.

2X correct mixture at formation determination buffer (Panvera), 10nMDTT and 40nMES2 on ice. 2X reactant mixture at also formation determination buffer (Panvera) and 20nMhER-β (Panvera) and 40nMES2 on ice.

The diluent of test compound is prepared with 30% (v/v) dimethyl sulfoxide/50mMHEPESpH7.5. Now, described diluent is final concentration needed for 40X.

Then in each hole, add 50 μ L correct mixtures. To porose middle addition 48 μ L reactant mixture. Diluted chemical compound liquid described in 2.5 μ L is added in appropriate well. Pipette mixes described reactant mixture by hand, covers on this plate by a rolls of aluminum foil adhesive faces, then this plate is incubated at room temperature 1 hour.

Then, on LjLAnalyst, read excitation wavelength 265nm and launch the reading in each hole of this plate under wavelength 538nm.

According to the method described above, testing the combination of representative compound of the present invention and estrogen receptor alpha and erss, result is shown in table 1.

Table 1

Embodiment 65:

MCF-7 cell proliferating determining

According to the method that (BreastCancetRes.Treat., 1987,10 (2), 169-75) such as Welshons are introduced, carry out this and measure.

MCF-7 cell maintaining RPMI1640 without in phenol red medium (Gibco), described culture medium contains 10%FBS (Hyclone), supplements bovine insulin and non essential amino acid (Sigma). Described cell is first with 4-hydroxyl tamoxifen (10^-8M) process, then allow it stand 24 hours in 37 DEG C. After carrying out this insulation with tamoxifen, the compound of described cell variable concentrations processes.

The testing compound of the Agonist Mode of variable concentrations is added in described culture medium. Prepare the pending compound of Antagonist Mode equally, in described culture medium, also add 10nM17 beta estradiol. Described cell is incubated 24 hours in 37 DEG C. After insulation, in described culture medium, add 0.1 μ Ci¹⁴C-thymidine (56mCi/mmol, Amersham), is incubated 24 hours by described cell then at 37 DEG C. Then described cell washes twice with HankShi buffer salt solution (HBSS) (Gibco), then counts with scintillation counter. Relative to Vehicle controls cell, with the cell of described compound treatment¹⁴C-thymidine increase is reported as cell proliferation increases percentage rate.

Measuring representative compound of the present invention according to the method described above, result is shown in table 2.

Table 2

NA represents and is not detected by activity under experimental concentration.

Embodiment 66

The alkaline phosphatase assay of people's endometrium Ishikawa cell

According to Albert etc., CancerRes, (9910), 50 (11), the method that 330-6-10 introduces carries out this and measures.

Ishikawa cell is maintained the DMEM/F12 (1:1) of supplementary 10% calf serum (Hyclone) without in phenol red medium (Gibco). Test first 24 hours, culture medium is replaced by containing 2% calf serum without phenol red DMEM/F12 (1:1).

The testing compound of the Agonist Mode of variable concentrations is added in described culture medium. Prepare the pending compound of Antagonist Mode equally, in described culture medium, also add 10nM17 beta estradiol. Described cell is incubated 3 days in 37 DEG C. At the 3rd day, remove culture medium, in each hole, be initially charged the 1X dilution buffer (Clontech) of 1 times of volume, add the mensuration buffer (Clontech) of 1 times of volume. Then described cell is incubated at room temperature 5 minutes. Add chemiluminescent buffer (chemical luminous substrate (CSPD) of 1 times of volume, the chemiluminescence intensifier of 19 times of volumes, the final concentration of 1.25mM of CSPD of the fresh preparation of 1 times of volume; SigmaChemicalCo.) described cell is incubated at room temperature 10 minutes, then carries out on luminometer quantitatively. Relative to Vehicle controls, increase the enhancing calculating alkaline phosphatase activities with chemiluminescence.

According to the method described above, testing representative compound of the present invention, result is shown in table 3.

Table 3

Embodiment 67: to the effect produced of IL-6 and GM-CSF in HOB cell

Human body osteoclast HOB bed board makes them in 96 hole wares, and in conventional H OB culture medium, (HamShi F12, is supplemented with 28mMHEPES, pH7.4,10%FCS, 1.1mMCaCl₂, 2mM glutamine and 1% Antibiotic-Antimycotic) in density be 7 × 103 cells/well. Next day, cell compound or vehicle treated (0.2%DMSO) process 30 minutes, are subsequently added IL-1 β (1ng/mL) and TNF-α (10ng/mL). Cultivate and continue 18 to 24 hours. Commercial ELISA Assay kit is utilized to measure IL-6 and GM-CSF level in culture medium. The compounds of this invention demonstrates the inhibitory action to IL-6 and GM-CSF.

Measuring representative compound of the present invention according to the method described above, result is shown in table 4.

Table 4

Embodiment 68: to the effect produced of IL-6 and GM-CSF in HOB cell

It is 7 × 10 that human body osteoclast HOB bed board makes its density in conventional H OB culture medium (HamShi F12 is supplemented with 28mMHEPES, pH7.4,10%FCS, 1.1mMCaCl2,2mM glutamine and 1% Antibiotic-Antimycotic) in 96 hole wares³Individual cells/well. Next day, cell compound or vehicle treated (0.2%DMSO) process 30 minutes, are subsequently added IL-1 β (1ng/mL) and TNF-α (10ng/mL). Cultivate and continue 18 to 24 hours. Commercial ELISA Assay kit is utilized to measure IL-6 and GM-CSF level in culture medium. The compounds of this invention demonstrates the inhibitory action to IL-6 and GM-CSF

Measuring representative compound of the present invention according to the method described above, result is shown in table 5.

Table 5

Embodiment 69: the effect to Proliferation of ovarian cancer cell SKOV 3

After exponential phase cell trypsinization, with 6 × 10³Individual/porocyte number adds people 96 well culture plate, puts 37 DEG C, 5%CO₂Incubator is cultivated, within the 2nd day, treats the adherent 4 DEG C of calorstat 1h of rearmounted people of most cells, to facilitate cell synchronization to grow. Suck supernatant, add people containing 10% newborn calf serum (FCS) RPMI1640 culture fluid, 200 μ L/ holes, empirically design packet. The compound injection liquid of physiological saline solution preparation is added in 96 holes, every hole adds 200 μ L, makes drug level respectively 1mg/mL, 2mg/mL and the 5mg/mI in every hole, with 0mg/mL for negative control group.Continue cultivation 24,48, after 72h, each hole adds people 20 μ LMTT solution (concentration is 5mg/mL) respectively, shake culture plate gently, put back to and in incubator, hatch 4h again, then exhaust supernatant, in each hole, add dimethyl sulfoxide 200 μ L, put concussion 5-10min on oscillator, measuring, with enzyme mark photometer, the light absorption value (A=580) that every hole medium wavelength is 580nm, A=580 value is directly proportional to living cells quantity.

Measuring representative compound of the present invention according to the method described above, result is shown in table 6.

Table 6

Embodiment 70: the effect to osteosarcoma U 2OS-EGFP-4A12G propagation

After exponential phase cell trypsinization, trypan blue counts, and being configured to cell density is 1 × l0⁴The cell suspension of individual/mL, is inoculated in 96 orifice plates, every hole 200 μ L, every hole about 2 × 10³Individual cell, preculture 24h, the compound injection liquid of physiological saline solution preparation is added in 96 holes, every hole adds 200 μ L, makes drug level respectively 1mg/mL, 2mg/mL and the 5mg/mI in every hole, with 0mg/mL for negative control group. After cultivating 0h, 12h, 24h and 48h respectively, every hole adds MTT solution (5mg/mL) 20 μ L, continues to hatch 4h, terminates cultivating. The supernatant in culture hole is abandoned in careful suction, and every hole adds the dimethyl sulfoxide (DMSO) of 150 μ L, shakes 10min, make the cured abundant dissolving of first, select 490nm wavelength, enzyme-linked immunosorbent assay instrument measures each hole absorbance value (A value), duplicate detection 5 times.

Measuring representative compound of the present invention according to the method described above, result is shown in table 7.

Table 7

Example of formulations

Following example of formulations protection scope of the present invention by way of example only, but constitute restriction never in any form.

Embodiment 71: gelatine capsule

The preparation of hard gelatin capsule adopts:

Above-mentioned preparation can be improved according to the reasonable change provided.

Embodiment 72: tablet

The preparation of tablet adopts

Said components is mixed and is pressed into tablet.

Embodiment 73: tablet

In every, the tablet preparation containing 2.5-1000mg active component is as follows:

Active component, starch and cellulose is made by No. 45 mesh sieves of the U.S. and to be thoroughly mixed. Polyvinylpyrrolidonesolution solution is mixed with gained powder, with after through No. 14 mesh sieves of the U.S.. The granule generated is dried and through No. 18 mesh sieves of the U.S. at 50-60 DEG C. Joining first passing through the sodium carboxymethyl cellulose of No. 60 mesh sieves of the U.S., magnesium stearate and Pulvis Talci in advance in above-mentioned granule, mix subsequently, on tablet machine, compacting obtains tablet.

Embodiment 74: suspension

The suspension preparation that every 5ml contains 0.1-1000mg medicine is as follows:

Medicine is made through No. 45 mesh sieves of the U.S. and to be mixed to form smooth paste with sodium carboxymethyl cellulose and syrup. Benzoic acid solution, correctives and coloring agent with the dilution of some water and are under agitation added aforesaid paste. It is subsequently added enough water to reach required volume.

Embodiment 75: combined tablet-preparation

For described above, those skilled in the art are readily understood by the essential feature of the present invention, and without departing substantially from the spirit and scope of the present invention, the present invention can carry out various changes and improvements to adapt to different application and condition.

Claims

1. following Isosorbide-5-Nitrae-dihydro-thiophene also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazole compound and pharmaceutically acceptable salt thereof, is selected from:

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-aminomethyl phenyl) piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(diphenyl methyl) piperazine;

The bromo-1,4-dihydro-thiophene of N, N-diethyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide;

1-(7-bromo-1,4-dihydro-thiophene also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(the chloro-4-fluorophenyl of 3-) piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-chlorphenyl) piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-Trifluoromethoxyphen-l) piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(2-fluorophenyl) piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-trimethoxyphenyl) piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-phenylpiperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-fluorophenyl) piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-methyl piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-benzyl diethylenediamine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-[(4-chlorphenyl) (phenyl) methyl] piperazine;

The bromo-1,4-dihydro-thiophene of the N-tert-butyl group-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(3-bromophenyl) piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(4-ethoxyl phenenyl) piperazine;

4-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl) morpholine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl) pyrrolidine;

The bromo-1,4-dihydro-thiophene of N-isopropyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide;

The bromo-1,4-dihydro-thiophene of N-(2-diethylin) ethyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide;

The bromo-1,4-dihydro-thiophene of N, N-dimethyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide;

The bromo-1,4-dihydro-thiophene of N-phenyl-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl) piperidines;

N (furan-2-methylene) the bromo-1,4-dihydro-thiophene of-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide;

N (pyridine-2-base) the bromo-1,4-dihydro-thiophene of-7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-amide;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(3,5-bis-trifluoromethyl) piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(3,4-difluorophenyl) piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(2-aminomethyl phenyl) piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(2-chlorphenyl) piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(2,5-3,5-dimethylphenyl) piperazine;

1-(the bromo-1,4-dihydro-thiophene of 7-also [3 ', 2 ': 5,6] thiapyran also [4,3-c] pyrazoles-3-carbonyl)-4-(3-trifluoromethyl) piperazine;

The bromo-1,4-dihydro-thiophene of the N tert-butyl group-7,8-two also [3', 2':5,6] thiapyran also [4,3-c] pyrazoles-3-amide.

2. following 3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazole compound and pharmaceutically acceptable salt thereof, is selected from:

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(benzhydryl phenyl) piperazine;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-chlorphenyl) piperazine;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-[(4-chlorphenyl) (phenyl) methyl] piperazine;

The N-tert-butyl group-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-methyl piperazine;

N, N-diethyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-aminomethyl phenyl) piperazine;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-phenylpiperazine;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-Trifluoromethoxyphen-l) piperazine;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-benzyl diethylenediamine;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(4-methoxyphenyl) piperazine;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(3-trifluoromethyl) piperazine;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(3-bromophenyl) piperazine;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(3,4-difluorophenyl) piperazine;

N, N-dimethyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl) pyrrolidine;

N-isopropyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide;

N-benzyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide;

N-phenyl-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide;

4-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl) morpholine;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl) piperidines;

N-(furan-2-methylene)-4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-amide;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(the chloro-4-fluorophenyl of 3-) piperazine;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(2-aminomethyl phenyl) piperazine;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(3,5-bis-trifluoromethyl) piperazine;

1-(4,5-dihydro-3aH-thieno [2', 3':2,3] thia also [4,5-c] pyrazoles-3-carbonyl)-4-(2-chlorphenyl) piperazine.

3. a pharmaceutical composition, comprises as compound described in any one of claim 1-2 of active component and pharmaceutically acceptable salt thereof and pharmaceutically acceptable carrier.

4. the compound described in claim 1 or 2 or the application in preparation treatment nonsmall-cell lung cancer, small cell lung cancer medicine of the pharmaceutical composition described in claim 3.

5. the compound described in claim 1 or 2 or the application in the disease medicament that preparation prevention is relevant to the imbalance of EGF-R ELISA signal transduction with treatment of the pharmaceutical composition described in claim 3.

6. application according to claim 5, it is characterised in that: described EGF-R ELISA is HER-1, HER-2, HER-3 or HER-4.

7. application according to claim 5, it is characterised in that: the relevant disease of wherein said EGF-R ELISA signal transduction imbalance is scale cancer, adenocarcinoma, large cell carcinoma, colorectal carcinoma, breast carcinoma, ovarian cancer, renal cell carcinoma or bronchial asthma.