WO2014044107A1

WO2014044107A1 - Pyrazol[3,4-c] pyridine derivative, preparation method and use in medicine thereof

Info

Publication number: WO2014044107A1
Application number: PCT/CN2013/082020
Authority: WO
Inventors: 许建烟; 吕贺军; 屈博磊; 董庆
Original assignee: 上海恒瑞医药有限公司; 江苏恒瑞医药股份有限公司
Priority date: 2012-09-18
Filing date: 2013-08-22
Publication date: 2014-03-27
Also published as: CN104039788A; WO2014044113A1; CN104039788B; TW201412737A

Abstract

The present invention relates to a pyrazol[3,4-c] pyridine derivative, preparation method and use in medicine thereof. Specifically, the present invention relates to a novel pyrazol[3,4-c] pyridine derivative, preparation method thereof and a pharmaceutical composition comprising the derivative and use thereof as a therapeutic agent, especially their use as Xa factor inhibitors and in the preparation of drugs for treating diseases like thromboembolism.

Description

Pyrazolo[3,4-c]pyridine derivatives, preparation method thereof and application thereof in medicine

Technical field

The present invention relates to a novel class of pyrazolo[3,4- _C ]pyridine derivatives, a process for the preparation thereof, and a pharmaceutical composition containing the same, and as a therapeutic agent, particularly as a factor Xa inhibitor and in the preparation of a therapeutic And use in drugs for preventing diseases such as thromboembolism. Background technique

Cardiovascular and cerebrovascular diseases are the first leading cause of death in the world today. Cardiovascular diseases include deep vein thrombosis (DVT), myocardial infarction (MI), pulmonary embolism (PE), and stroke caused by atrial fibrillation. Thrombosis is caused by myocardial infarction, stroke, deep vein thrombosis, pulmonary embolism, and myocardial infarction. Important causes of cardiovascular diseases such as stroke, stroke and atrial fibrillation. According to data released by the American Heart Association in 2009, 278.9 people died of cardiovascular disease per 10,000 Americans in 2005. In 2006, a total of 2,425,900 out of 900,072 (34.2%) of the 900 deaths were caused by cardiovascular death. One in 2.9 patients died of cardiovascular disease.

When the vascular system of a key organ is blocked by a blood clot, it can cause thromboembolism, affecting the normal supply of oxygen and nutrients. When the blood clot develops to block the arterial tube, it is very likely to cause a stroke or heart disease. The incidence and mortality of venous thromboembolism (VTE) is extremely high. Every year, the number of VTEs in the EU exceeds 1 million, and the number of deaths per year is more than 500,000. In the United States, the number of VTEs per year exceeds 600,000 and the death toll is nearly 300,000.

Standard VTE treatments include vitamin K antagonists such as warfarin combined with unfractionate heparin (UFH) or low-molecular-weight heparin (LMWH). However, this treatment has a large number of Restricted, warfarin is administered orally, but the initial effect is slow, the individual difference is large, the treatment window is narrow, and the food and drug are affected greatly. It is necessary to constantly monitor the international standard clotting time ratio (0NR), which may increase the cost of treatment; Significant unfractionated heparin can only be administered parenterally, the anticoagulant effect is difficult to predict, and life-threatening heparin-induced thrombocytopenia is associated with a high risk of bleeding, requiring strict patient monitoring. Due to the above limitations, anticoagulants which selectively inhibit a single specific coagulation factor have recently been developed and can be classified into direct thrombin inhibitors (DTIs) and coagulation factor Xa inhibitors.

Normally, coagulation proteins are present in the blood in an inactive state. When the blood vessels are damaged, a coagulation cascade is triggered, which eventually leads to the formation of fibrin clots, which form a thrombus after stabilization. The coagulation process includes two major pathways: endogenous (; maintenance) and exogenous. When the intima of the vessel ruptures, the factor Xn (factor Xn, fXII) in the serum is activated in the presence of kininase to form factor XIIa (factor Xlla, EXIIa), and EXIIa catalyzes the activation of the DQ in the activated state of the £XIa shear downstream. The flX produces active flXa, which binds to Villa. In the presence of Ca ions and phospholipids, it is activated by thrombin or £Xa to form a complex called prothrombin activator, which in turn activates factor X (fa _C tor X, fX), forming £Xa. Prothrombin activator catalytic factor II formation The active ffla hydrolyzes the downstream factor I to form water-insoluble fibrin to form a blood clot. The factors involved in this coagulation process are all in serum and are therefore referred to as endogenous pathways. Exogenous coagulation is initiated by tissue damage release factor ,. When the blood vessel ruptures, tissue factor III forms a complex with activated fVIIa in the serum, and activates in the presence of Ca ^{2+ to} become £Xa £Xa. The process of exogenous coagulation and endogenous coagulation is consistent.

In summary, the coagulation cascade is a process in which a series of proteases are gradually activated by exogenous (tissue factor, factor Vila) and endogenous (cause XIIa XIa, IXa and Villa) pathways. The cascade reaction eventually aggregates into factor Xa to form a common pathway, and thrombin (factor Ila) plays a central role in the coagulation reaction, so both are the main targets for anticoagulant drugs. Xa factor inhibitors selectively inhibit factor Xa, prolong clotting time, and reduce thrombin generation to achieve antithrombotic effects.

A series of patent applications for Factor Xa inhibitors are disclosed, including WO2001047919 WO2008006479 WO2007137801 or WO2006047528 and the like.

Although a series of anti-thrombotic Factor Xa inhibitors have been disclosed, there is still a need to develop new compounds with better pharmacodynamics. Through continuous efforts, the present invention has devised a series of new compounds and found these compounds. Shows excellent results and effects. Summary of the invention

The object of the present invention is to provide a compound as shown in Table 1 or a tautomer, a mesogen, a racemate, an enantiomer, a diastereomer thereof and a mixture thereof, and Pharmaceutically acceptable salt:

Table 1

Example No. Compound Structure and Nomenclature

1

1-(2-Fluoro-4-methoxyphenyl)-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro Pyrazolo[3,4- _c ]pyridine-3-carboxamide

2 6 ^U

1-(4-methoxyphenyl)-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-4,5,6,7-tetrahydropyrazole

[3,4- _c ]pyridine-3-carboxamide

The present invention also relates to a compound shown below or a tautomer, a mesogen, a racemate, an enantiomer, a diastereomer, a mixture thereof, and a pharmaceutically acceptable form thereof Salt:

These compounds can be used as intermediates for the synthesis of the corresponding compounds in Table 1, wherein: R ¹ is an alkyl group; preferably a d- ₆ alkyl group, more preferably a d- ₄ alkyl group, most preferably an ethyl group.

Further, another aspect of the invention relates to a pharmaceutical composition comprising a therapeutically effective amount of the compound shown in Table 1 or a tautomer, racemate, enantiomer, diastereomer thereof In the form of a mixture, a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier, diluent or excipient thereof.

Another aspect of the invention relates to the compounds shown in Table 1, or the tautomers, racemates, enantiomers, diastereomers, mixtures thereof, and pharmaceutically acceptable salts thereof, Or use of a pharmaceutical composition comprising the same for the preparation of a prophylactic and/or therapeutic thromboembolic disorder. The disease is selected from the group consisting of cardiac arrest, angina pectoris, angioplasty or aortic coronary artery shunt re-occlusion and restenosis, stroke, transient ischemic attack, peripheral arterial occlusive disease, pulmonary embolism or deep vein Thrombosis.

Another aspect of the invention relates to the compounds shown in Table 1, or the tautomers, racemates, enantiomers, diastereomers, mixtures thereof, and pharmaceutically acceptable salts thereof, Or use of a pharmaceutical composition comprising the same for the preparation of a medicament for preventing and/or treating a disease which is positively affected by inhibition of factor Xa.

Another aspect of the invention relates to the compounds shown in Table 1, or the tautomers, racemates, enantiomers, diastereomers, mixtures thereof, and pharmaceutically acceptable salts thereof, Or use of a pharmaceutical composition comprising the same in the manufacture of a medicament for the treatment of disseminated intravascular coagulation.

Another aspect of the invention relates to the compounds shown in Table 1, or the tautomers, racemates, enantiomers, diastereomers, mixtures thereof, and pharmaceutically acceptable salts thereof, Or use of a pharmaceutical composition comprising the same in the preparation of a medicament for inhibiting Factor Xa. Another aspect of the invention relates to a method of modulating inhibitory factor Xa activity, preferably inhibiting factor Xa activity, comprising administering to a subject in need thereof a therapeutically effective amount of a compound shown in Table 1 or a tautomer thereof, Racemates, enantiomers, diastereomers, mixtures, and pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the same.

Another aspect of the invention relates to a method of treating or preventing the preparation of a prophylactic and/or therapeutic thromboembolic disorder, the method comprising administering to a patient in need of treatment a therapeutically effective amount of a compound of Table 1 or a tautomer thereof , a racemate, an enantiomer, a diastereomer, a mixture, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same. The disease is selected from the group consisting of cardiac arrest, angina pectoris, angioplasty or aortic coronary artery shunt re-occlusion and restenosis, stroke, transient ischemic attack, peripheral arterial occlusive disease, pulmonary embolism or deep vein Thrombosis.

Another aspect of the invention relates to a method for the preparation of a medicament for preventing and/or treating a positive effect by inhibiting factor Xa, which comprises administering to a patient in need of treatment a therapeutically effective amount of the compound shown in Table 1 or its interconversion An isomer, a racemate, an enantiomer, a diastereomer, a mixture, a pharmaceutically acceptable salt thereof, or a pharmaceutical composition comprising the same.

Another aspect of the invention relates to a method of preparing a treatment for disseminated intravascular coagulopathy comprising administering to a patient in need of treatment a therapeutically effective amount of a compound shown in Table 1 or a tautomer thereof, a racemate thereof , enantiomers, diastereomers, mixtures, and pharmaceutically acceptable salts thereof, or pharmaceutical compositions comprising the same. Detailed description of the invention

Unless otherwise stated, the following terms used in the specification and claims have the following meanings. "Alkyl" means a saturated aliphatic hydrocarbon group including straight chain and branched chain groups of 1 to 20 carbon atoms. An alkyl group having 1 to 10 carbon atoms is preferred, and an alkyl group having 1 to 6 carbon atoms is more preferred. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, tert-butyl, sec-butyl, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethylpropyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2 -methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethylbutyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1, 3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, 2,3-dimethylbutyl, n-heptyl, 2-methylhexyl, 3-methylhexyl, 4-methylhexyl, 5-methylhexyl, 2,3-dimethylpentyl, 2,4-dimethylpentyl, 2,2-dimethyl Pentyl, 3,3-dimethylpentyl, 2-ethylpentyl, 3-ethylpentyl, n-octyl, 2,3-dimethylhexyl, 2,4-dimethylhexyl, 2,5-Dimethylhexyl, 2,2-dimethylhexyl, 3,3-dimethylhexyl, 4,4-dimethylhexyl, 2-ethylhexyl, 3-ethylhexyl, 4- Ethylhexyl, 2-methyl-2-ethylpentyl, 2- Methyl-3-ethylpentyl, n-decyl, 2-methyl-2-ethylhexyl, 2-methyl-3-ethylhexyl, 2,2-diethylpentyl, n-decyl, 3,3-Diethylhexyl, 2,2-diethylhexyl, and various branched isomers thereof. More preferred are lower alkyl groups having 1 to 6 carbon atoms, and non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, t-butyl, sec-butyl Base, n-pentyl, 1,1-dimethylpropyl, 1,2-dimethylpropyl, 2,2-dimethylpropyl, 1-ethyl Propyl, 2-methylbutyl, 3-methylbutyl, n-hexyl, 1-ethyl-2-methylpropyl, 1,1,2-trimethylpropyl, 1,1-dimethyl Butyl, 1,2-dimethylbutyl, 2,2-dimethylbutyl, 1,3-dimethylbutyl, 2-ethylbutyl, 2-methylpentyl, 3- Methyl amyl, 4-methylpentyl, 2,3-dimethylbutyl, and the like. The alkyl group may be substituted or unsubstituted, and when substituted, the substituent may be substituted at any available point of attachment, preferably one or more of the following groups, independently selected from alkyl, alkenyl, Block group, alkoxy group, alkylthio group, alkylamino group, halogen, thiol, hydroxy group, nitro group, cyano group, cycloalkyl group, heterocycloalkyl group, aryl group, heteroaryl group, cycloalkoxy group, hetero Cycloalkoxy, cycloalkylthio, heterocycloalkylthio, oxo.

"Optional" or "optionally" means that the event or environment described subsequently may, but need not, occur, including where the event or environment occurs or does not occur. For example, "heterocyclic group optionally substituted by an alkyl group" means that an alkyl group may be, but not necessarily, present, including the case where the heterocyclic group is substituted by an alkyl group and the case where the heterocyclic group is not substituted by an alkyl group. .

"Substituted" means one or more hydrogen atoms in the group, preferably up to 5, more preferably 1 to 3, hydrogen atoms are independently substituted with each other by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and those skilled in the art will be able to determine (by experiment or theory) substitutions that may or may not be possible without undue effort. For example, an amino or hydroxyl group having a free hydrogen may be unstable when combined with a carbon atom having an unsaturated (e.g., olefinic) bond.

"Pharmaceutical composition" means a mixture containing one or more of the compounds described herein, or a physiologically/pharmaceutically acceptable salt or prodrug thereof, and other chemical components, as well as other components such as physiological/pharmaceutically acceptable carriers. And excipients. The purpose of the pharmaceutical composition is to promote the administration of the organism, which facilitates the absorption of the active ingredient and thereby the biological activity. DRAWINGS

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a graph showing the comparison of the APTT parameters of the compound of Example 1 of the present invention with APIXABAN.

Figure 2 is a graph showing the comparison of the PT parameters of the compound of Example 1 of the present invention with APIXABAN. detailed description

Example

The structure of the compound is determined by nuclear magnetic resonance (1H NMR) and/or mass spectrometry (MS). The iHNMR shift (δ) is given in parts per million (ppm). The 1H NMR measurement was performed on a Bruker AVANCE-400 nuclear magnetic apparatus, and the solvent was deuterated methanol (CD ₃ OD), deuterated chloroform (CDC1 ₃ ), hexamethyl dimethyl sulfoxide (OMSO-d ₆ ), internal standard. It is tetramethylsilane (TMS).

The MS was assayed using a FINMGAN LCQAd (ESI) mass spectrometer (manufacturer: Thermo, model: Finnigan LCQ advantage MAX).

The HPLC was measured using an Agilent 1200 DAD high pressure liquid chromatograph (Sunfire C 18 150 x 4.6 mm column) and a Waters 2695-2996 high pressure liquid chromatograph (Gimini C18 150 x 4.6 mm column).

The IC ₅ o value was determined using a NovoStar plate reader (BMG, Germany). Thin layer chromatography silica gel plate using Yantai Yellow Sea HSGF254 or Qingdao GF254 silica gel plate, thin layer chromatography

(TLC) The silica gel plate used has a specification of 0.15 mm to 0.2 mm, and the thin layer chromatography separation and purification product has a specification of 0.4 mm to 0.5 mm.

The silica gel column generally uses Yantai Huanghai silica gel 200~300 mesh silica gel as a carrier.

The alkaline alumina column is generally used as a carrier for FCP200~300 mesh basic alumina using the national medicine chromatography.

Known starting materials of the invention may be synthesized or synthesized according to methods known in the art, or may be

Purchased by ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc and Dari Chemicals.

Unless otherwise stated in the examples, the reactions were all carried out under a nitrogen or argon atmosphere.

An argon atmosphere or a nitrogen atmosphere means that the reaction flask is connected to an argon or nitrogen balloon having a volume of about 1 L.

The hydrogen atmosphere means that the reaction flask is connected to a hydrogen balloon of about 1 L volume.

The pressurized hydrogenation reaction uses a Parr Model 3916EKX hydrogenation apparatus and a clear blue QL-500 hydrogen generator or a HC2-SS type hydrogenation apparatus.

The hydrogenation reaction is usually evacuated, charged with hydrogen, and operated three times.

There is no particular description in the examples, and the solution means an aqueous solution.

Unless otherwise specified in the examples, the reaction temperature is room temperature and is 20 ° C to 30 ° C.

The progress of the reaction in the examples was monitored by thin layer chromatography (TLC). The systems used for the reaction were: dichloromethane and methanol systems, n-hexane and ethyl acetate systems, petroleum ether and ethyl acetate systems, The volume ratio of acetone to solvent is adjusted depending on the polarity of the compound.

The system for the elution of the column chromatography and the system for the thin layer chromatography of the developer used for the purification of the compound include: A: dichloromethane and methanol systems, B: n-hexane and ethyl acetate systems, C: dichloromethane and Acetone system, D: methanol, E: petroleum ether and ethyl acetate, F: ethyl acetate and methanol, the volume ratio of the solvent is adjusted according to the polarity of the compound, and a small amount of triethylamine or the like may be added or The acid reagent such as acetic acid is adjusted.

Example 1

(2-fluoro-4-methoxyphenyl)-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro-1H -pyrazole

[3,4-c]pyridine-3-carboxamide

First step

2-Chloro-2-(2-(2-fluoro-4-methoxyphenyl)hydrazinyl)ethyl acetate 2-fluoro-4-methoxyaniline la (10.20 g, 72.30 mmol) was added to 30 In mL water, at 0 ° C, 18 mL of concentrated hydrochloric acid and 12.7 mL of 40% aqueous sodium nitrite were slowly added dropwise, and the reaction was stirred at 0 ° C for 1.5 hours, and stored at 0 ° C as a reserve liquid 1. Ethyl 2-chloroacetoacetate (12.90 g, 78.80 mmol) and sodium acetate (11.80 g, 144.60 mmol) were added to a mixed solution of 50 mL of acetone and water < /V=2:3), and added at 0 °C. Liquid 1, after completion of the dropwise addition, the reaction was stirred at 0 ° C for 1 hour and at 0 ° C for 12 hours. 200 mL of dichloromethane and 20 mL of methanol were added to the reaction solution, and the mixture was stirred for 3 hours under ice-cold water, and the mixture was stirred at room temperature. The reaction was stirred for 20 hours, and the aqueous phase was extracted with 200 mL of dichloromethane. Wash with water (150 mL) and EtOAc (EtOAc m. Methoxyphenyl)hydrazinyl)ethyl acetate lb (20.10 g, yellow solid), Yield: 100%.

Second step

(2-fluoro-4-methoxyphenyl)-6-(4-nitrophenyl)-7-carbonyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4- _C ]pyridine

Ethyl 3-carboxylate

3-Chloro-1-(4-nitrophenyl)-5,6-dihydropyridine-2 (1H ketone lc (9 g, 36 mmol, using well-known and 2-chloro-2-(2-) 2-fluoro-4-methoxyphenyl)hydrazinyl)ethyl acetate lb (9.75 g, 36 mmol) was dissolved in 150 mL of toluene, triethylamine (11 g, 107 mmol), and stirred at 90 ° C The reaction was carried out for 12 hours. 200 mL of dichloromethane and 200 mL of water were added to the reaction mixture, and the mixture was separated. The aqueous phase was extracted with dichloromethane (200 mL X 3), and the organic phase was combined with saturated sodium chloride solution (200 mL) The organic layer was dried (MgSO4) , 7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester ld (8.40 g, pale yellow solid), Yield: 51%.

6- 4-Aminophenyl) Fluoro-4-methoxyphenyl)-7-carbonyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine

Ethyl 3-carboxylate

1-(2-Fluoro-4-methoxyphenyl)-6-(4-nitrophenyl)-7-carbonyl-4,5,6,7-tetrahydro-1H-pyrazolo[3 4- _C ] Pyridine-3-carboxylic acid ethyl ester Id (6 g, 13.20 mmol) was dissolved in 120 mL of methanol, 10% palladium carbon (1.20 g, 20%) was added, and hydrogen was replaced three times, and the reaction was stirred for 1 hour. Filtration and concentration of the filtrate under reduced pressure afforded title product 6-(4- Aminophenyl)-l-(2-fluoro-4-methoxyphenyl)-7-carbonyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4- _C ]pyridine- Ethyl 3-carboxylate (5.30 g, yellow solid), Yield: 94.6%.

the fourth step

_6- (4-(5-bromopentylamide)phenyl)fluoro-4-methoxyphenyl 7-carbonyl-4,5,6,7-tetrahydro-1H-pyrazole

Ethyl [3,4- _C ]pyridine-3-carboxylate

6-(4-Aminophenyl)-1-(2-fluoro-4-methoxyphenyl)-7-carbonyl-4,5,6,7-tetrahydro-1H-pyrazolo[3, 4- _C ] pyridine-3-carboxylic acid ethyl ester le (5.30 g, 12.50 mmol) and triethylamine (3.80 g, 37.50 mmol) were dissolved in 100 mL of dichloromethane, and 10 mL of 5-bromopentane was added dropwise at 0 °C. A solution of the acid chloride (3.20 g, 16.20 mmol) in dichloromethane was added dropwise, and the mixture was warmed to room temperature, and the reaction was stirred for 3 hours. 50 mL of water was added to the reaction mixture, and the mixture was extracted with methylene chloride (50 mL EtOAc). Filtration gave the title product 6-(4-(5-bromopentyl)phenyl)-1-(2-fluoro-4-methoxyphenyl)-7-carbonyl-4,5,6,7-tetra Hydrogen-1H-pyrazolo[3,4- _C ]pyridine-3-carboxylic acid ethyl ester If (7.35 g, oil), Yield: 100%.

the fifth step

1-(2-Fluoro-4-methoxyphenyl)-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro -1H-pyrazole

Ethyl [3,4- _C ]pyridine-3-carboxylate

_6- (4-(5-Bromopentylamide)phenyl)-1-(2-fluoro-4-methoxyphenyl)-7-carbonyl-4,5,6,7-tetrahydro-1H- Ethyl pyrazolo[3,4-c]pyridine-3-carboxylate If (7.35 g, 12.50 mmol) was dissolved in 100 mL of tetrahydrofuran, and potassium t-butoxide (1.70 g, 15 mmol) was added at 0 ° C. The reaction was stirred for 1 hour at room temperature. 50 mL of a saturated ammonium chloride solution was added to the reaction mixture, and the mixture was extracted with ethyl acetate (50 mL×3). The organic phase was combined, washed with saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate The residue was purified by silica gel column chromatography elut elut elut elut elut elut elut Ethyl carbonyl piperidin-1-yl)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylate lg (3.90 g, yellow solid) , Yield: 61.9%.

Step 6

[3,4-c]pyridine-3-carboxamide

1-(2-Fluoro-4-methoxyphenyl)-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetra Hydrogen-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester lg (2.50 g, 5 mmol) dissolved in 60 mL of formamide and hydrazine, hydrazine-dimethylformamide (V/V = l :5), add 5 mL of trimethyl orthoformate and trifluoroacetic acid (0.10 g, 1 mmol) at 50 ° C, stir the reaction at 50 ° C for 30 minutes, add 5 mL of sodium methoxide (2.70 g, 25 mmol) The methanol solution was stirred at 50 ° C for 1.5 hours. 150 mL of water was added to the reaction mixture, and extracted with dichloromethane (200 mL X 2 ). The organic phase was combined, washed with saturated sodium chloride solution (100 mL X 3 ), dried over anhydrous sodium sulfate, filtered, filtrate reduced The residue was purified by silica gel column chromatography eluting elut elut elut eluting -carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxamide 1 (1.70 g, yellow solid) , the yield was 70.8%. MS m/z (ESI): 478.4 [M+l]

1H NMR (400 MHz, CDC1 ₃ ) δ 7.39 (t, 1H), 7.34 (d, 2H), 7.24-7.26 (m, 2H), 6.82 (s, 1H), 6.71-6.78 (m, 2H), 5.43 (s, 1H), 4.13 (t, 2H), 3.82 (s, 3H), 3.57-3.62 (m, 2H), 3.38 (t, 2H), 2.55-2.60 (m, 2H), 1.91-1.96 (m , 4H). Example 2

L-(4-Methoxyphenyl)-7-carbonyl-6-C4-0carbonylmorpholine)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4- c]pyridyl

4- 4-iodophenyl)morpholine-3-one

Morpholin-3-one 2a (0.46 g, 4.99 mmol, using well-known method literature)

Communications (Cambridge, United Kingdom), 2012, 48(1), 145-147 "prepared" and 1,4-diiodobenzene 2b (3 g, 9.09 mmol) dissolved in 5 mL of dioxane, in turn Iodinide (0.04 g, 0.23 mmol), cesium carbonate (4.44 g, 13.60 mmol) and N,N'-dimethylethylenediamine (0.08 g, 0.91 mmol) were added, and the reaction was stirred at 90 ° C for 12 hours. Filtration, the filtrate was concentrated under reduced pressure. EtOAcjjjjjjjjjj , Yield: 30%.

Second step

4-(4-(3-morpholine-2-carbonyl-5,6-dihydropyridine-1(2H)phenyl)morpholine-3-one

3-morpholine-5,6-dihydropyridine-2 (1H ketone 2d (419 mg, 2.31 mmol, prepared by the method disclosed in "Patent Application WO2004083177") and 4-(4-iodophenyl)morpholine 3-ketone 2c (700 mg, 2.31 mmol) was dissolved in 10 mL of dioxane, followed by iodide (21.90 mg, 0.12 mmol), cesium carbonate (2.20 g, 6.90 mmol) and N, N' - dimethylethylenediamine (40 mg, 0.46 mmol), refluxed for 40 hours. The filtrate was concentrated under reduced pressure. -0 morpholin-2-carbonyl-5,6-dihydropyridine-1 C2H)-yl)phenyl)morpholin-3-one 2e (370 mg, yellow solid), yield: 36.5%. third step

L-(4-Methoxyphenyl)-7α-morpholine-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-3α,4,5,6,7,7β-six Hydrogen-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester

4-(4-(3-Morpholine-2-carbonyl-5,6-dihydropyridine-1(2H)phenyl)morpholin-3-one 2e (357 mg, 1 mmol) and (Z) 2-Chloro-2-(2-(4-methoxyphenyl)phosphonium)acetate 2f (256 mg, 1 mmol, using a known method "3⁄4^, 2011 (2d), (11) , prepared by J 7 SOT) dissolved in 20 mL of ethyl acetate, added with triethylamine (505 mg, 5 mmol), refluxed for 4 hours, and stirred at 60 ° C for 12 hours. 20 mL of water was added to the reaction solution. The mixture was extracted with ethyl acetate (30 mL EtOAc). The resulting residue was purified to give the title product 1-(4-methoxyphenyl)-7α-morpholin-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-3? Ethyl 4,5,6,7,7?-hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylate 2 g (370 mg, yellow solid). Yield: 64.1%.

the fourth step

1-(4-Methoxyphenyl)-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3 , 4-c] ethylpyridin-3-carboxylate

1-(4-Methoxyphenyl)-7α-morpholine-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-3α,4,5,6,7,7β- Ethyl hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylate 2g (200 mg, 0.35 mmol) was dissolved in 5 mL of tetrahydrofuran, 0.9 mL of 4 M hydrochloric acid was added, and the reaction was stirred at 0 °C. hour. The saturated sodium bicarbonate solution was added dropwise to the reaction mixture to give a mixture of EtOAc (EtOAc) (EtOAc) (EtOAc) Filtration and concentration of the filtrate under reduced pressure afforded the title product 1-(4-methoxyphenyl)-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl) -4,5,6,7 Ethyl tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylate 2 h (200 mg, yellow solid). Yield: 100%.

the fifth step

1-(4-Methoxyphenyl)-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3 , 4-c] guanidin-3-carboxamide

1-(4-Methoxyphenyl)-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[ Ethyl 3,4-c]pyridine-3-carboxylate 2h (200 mg, 0.35 mmol) was dissolved in 10 mL of N,N-dimethylformamide, followed by 2 mL of formamide and 2 mL of trimethyl orthoformate Stir the reaction with 6 trifluoroacetic acid at 50 ° C for 30 minutes. 6 mL of sodium methoxide C189 mg, 3.50 mmol) in methanol was added, and the reaction was stirred at 50 ° C for 5 hours. The reaction mixture was poured into water (10 mL), EtOAc (EtOAc (EtOAc) The residue obtained was purified by EtOAc (EtOAc) (EtOAc) -4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxamide 2 (40 mg, yellow solid), yield: 25%.

1H NMR (400 MHz, CDC1 ₃ ) δ 7.48 (d, 2H), 7.33-7.39 (m, 4H), 6.94 (d, 2H), 6.84 (s, 1H), 5.45 (s, 1H), 4.34 (s , 2H), 4.12 (t, 2H), 4.03 (t, 2H), 3.83 (s, 3H), 3.73 (t, 2H), 3.39 (t, 2H). Example 3

L-(6-Methoxypyridin-3-yl)-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro- 1H-pyrazole

First step

(Z)-2-Chloro-2-(2-(6-methoxypyridin-3-yl)hydrazinyl)ethyl acetate 3-morpholine-1-(4-(2-carbonylpiperidine-1) -yl)phenyl)-5,6-dihydropyridine-2 (1H ketone 6-methoxypyridin-3-amine 3a (2 g, 16 mmol) was added to 8 mL of water at 0 ° C, slowly dropping Add 3.9 mL of concentrated hydrochloric acid and 10 mL of sodium nitrite (1.33 g, 19 mmol) in an aqueous solution, stir at 0 °C for 2 hours, and store as a stock solution 1. Dissolve sodium acetate C 3.03 g, 37 mmol) in 20 mL In the water, add 16 mL of acetone and ethyl 2-chloroacetoacetate (2.65 g, 16 mmol), add the stock solution 1 at 0 ° C, drip, stir the reaction at 0 ° C for 3 hours, add 100 mL of dichloromethane to stand. 12 hours. The liquid phase was extracted with dichloromethane (200 mL EtOAc). EtOAc (EtOAc) The residue obtained was purified by silica gel column chromatography eluting elut elut Ethyl acetate 3b (3.40 g, yellow solid), Yield: 72.3%

Second step

1-(6-Methoxymorpholin-3-yl)-7α-morpholine-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-3α,4,5 ,6,7,7β-hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester

(Z)-2-Chloro-2-(2-(6-methoxypyridin-3-yl)hydrazinyl)acetate 3b (86.90 mg, 0.34 mmol) and 3-morpholin-1-( 4-(2-Carbopiperidin-1-yl)phenyl)-5,6-dihydropyridine-2 (1H ketone 3c (120 mg, 0.34 mmol, obtained by the method disclosed in "Patent Application WO2003049681") Dissolved in 10 mL of acetic acid To the ester, triethylamine (102.20 mg, 1.01 mmol) was added and the reaction was refluxed for 22 hours. The reaction mixture was washed with water (10 mL), EtOAc (EtOAc) The title product 1-(6-methoxymorpholin-3-yl)-7α-morpholine-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-3α,4 , 5,6,7,7β-Hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 3d (100 mg, colorless oil), yield: 51%.

MS m/z (ESI): 577.3 [M+l]

third step

Ethyl [3,4- _C ]pyridine-3-carboxylate

1-(6-Methoxymorpholin-3-yl)-7α-morpholin-7-carbonyl-6-C4-C2-carbonylpiperidin-1-yl)phenyl)-3β,4,5, 6,7,7β-hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 3d (100 mg, 0.17 mmol) was dissolved in 5 mL of tetrahydrofuran, and 210 was added at 0 °C. 4M hydrochloric acid, stirred for 2 days. Add saturated sodium bicarbonate solution to the reaction solution ρΗ 7~8, add water ClO mL), extract with dichloromethane (15 mL X 3), combine the organic phases, and then use water (10 mL) and saturated sodium chloride. The solution was washed with EtOAc (EtOAc) (EtOAcjHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH Carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 3e (100 mg, colorless oil ()), Yield: 100%.

MS m/z (ESI): 490.2 [M+l]

the fourth step

[3,4-c]pyridine-3-carboxamide

1-(6-Methoxypyridin-3-yl)-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro -1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 3e (100 mg, 0.20 mmol) was dissolved in 3 mL of N,N-dimethylformamide, and then 1 mL of formamide was added. l mL of trimethyl orthoformate and 3 L of trifluoroacetic acid were stirred at 50 ° C for 30 minutes. 1.5 mL of sodium methoxide (32 mg, 0.59 mmol) in methanol was added and the reaction was stirred at 50 ° C for 5 hours. The reaction mixture was poured into 10 mL of water and extracted with methylene chloride (15 mL EtOAc). The organic layer was washed with water (15 mL) and saturated sodium chloride (15 mL) The filtrate was concentrated under reduced pressure, and the obtained residue was purified to purified crystals to afford to afford to afford the title product 1-(6-methoxypyridin-3-yl)-7-carbonyl-6-(4-( 2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxamide 3 (30 mg, white solid ), Yield: 31.8%.

1H NMR (400 MHz, DMSO-d6) δ 8.40 (d, 1H), 7.94 (dd, 1H), 7.76 (s, 1H), 7.48 (s, 1H) _: 7.38 (d, 2H), 7.27 (d, 2H), 6.91 (d, 1H), 4.05 (t, 2H), 3.89 (s, 3H), 3.58 (t, 2H), 3.21 (t, 2H), 2.38 (t, 2H), 1.78-1.90 (m , 4H). Example 4

lO chlorophenyl)-7-carbonyl-6-C4-0carbonylmorpholine)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3- Formamide

4c

First step

L-(3-Chlorophenyl)-7α-morpholine-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-3β,4,5,6,7,7β-hexahydro- 1H-pyrazole

Ethyl [3,4- _C ]pyridine-3-carboxylate

4-(4-(3-Morpholine-2-carbonyl-5,6-dihydropyridine-1(2H)phenyl)morpholin-3-one 2e (300 mg, 0.84 mmol) and (Z) 2-Chloro-2-(2-(3-chlorophenyl)hydrazinyl)acetate 4a (172 mg, 0.84 mmol, obtained by a known method "2011, (11), J 7 SOT) Dissolve in 20 mL of ethyl acetate, add triethylamine (424 mg, 4.20 mmol), and stir the reaction for 12 hours at 75 ° C. Add 30 mL of water to the reaction solution, and extract with ethyl acetate (20 mL X 3 ). The organic phase was washed with EtOAc EtOAc (EtOAc) -6-(4-(3-carbonylmorpholine)phenyl)-3β,4,5,6,7,7β-hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid Ester 4b (489 mg, pale yellow solid), Yield: 100%.

Second step

1-(3-Chlorophenyl)-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4 -c]pyridine-3-carboxylic acid ethyl ester

1-0 chlorophenyl)-7α-morpholine-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-3β,4,5,6,7,7β-hexahydro-1H - Pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 4b (489 mg, 0.84 mmol) was dissolved in 20 mL of tetrahydrofuran, added 2.2 mL of 4 M hydrochloric acid, and stirred at 0 ° C for 2 hours, 0 ° C was allowed to stand for 48 hours. Add saturated sodium bicarbonate solution to the reaction solution ρ Η 8, add water C 30 mL) and dichloromethane C 30 mL), separate the liquid, extract the aqueous phase with dichloromethane (30 mL X 2), combine the organic phase, and saturate The sodium chloride solution (50 mL) was washed with EtOAc (EtOAc m. )-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-4,5,6,7-tetrahydro-1H-pyrazole Ethyl [3,4-c]pyridine-3-carboxylate 4c (200 mg, yellow solid), yield: 100%.

third step

1-(3-Chlorophenyl)-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3, 4-c] Pyridine-3-carboxylic acid ethyl ester 4c (150 mg, 0.30 mmol) was dissolved in 10 mL of N,N-dimethylformamide, followed by 2.8 mL of formamide, 1 mL of trimethyl orthoformate and 4.6 L-trifluoroacetic acid was stirred at 50 ° C for 30 minutes. 1.2 mL of sodium methoxide C160 mg, 2.96 mmol) in methanol was added and the reaction was stirred at 50 ° C for 5 hours. The reaction mixture was poured into water (20 mL), EtOAc (EtOAc m. The residue obtained was purified by silica gel column chromatography eluting elut elut elut elut elut elut elut 4,5,6,7-Tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxamide 4 (50 mg, pale yellow solid), Yield: 35%.

1H NMR (400 MHz, CDC1 ₃ ) δ 7.61 (dd, 1H), 7.49 (ddd, 1H), 7.34-7.41 (m, 6H), 6.84 (s, 1H), 5.51 (s, 1H), 4.34 (s , 2H), 4.14 (t, 2H), 4.03 (t, 2H), 3.74 (t, 2H), 3.40 (t, 2H). Example 5

1-O-fluoro-4-methoxyphenyl)-7-carbonyl-6-C4-0carbonylmorpholine)phenyl)-4,5,6,7-tetrahydro-1H-pyrazole

1-(2-Fluoro-4-methoxyphenyl)-7α-morpholine-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-3α,4,5,6,7 ,7β-hexahydrogen

-1H-pyrazolo[3,4-c]pyridine-3-carboxylate

4-(4-(3-Morpholine-2-carbonyl-5,6-dihydropyridine-1(2H)phenyl)morpholin-3-one 2e (300 mg, 0.84 Methyl acetate lb (231 mg, 0.84 mmol) of 2-chloro-2-(2-(2-fluoro-4-methoxyphenyl)hydrazinyl) was dissolved in 20 mL of ethyl acetate. Triethylamine (424 mg, 4.20 mmol) was stirred at 75 °C for 12 hours. To the reaction mixture was added 50 mL of EtOAc (EtOAc)EtOAc. The title product 1-(2-fluoro-4-methoxyphenyl)-7α-morpholin-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-3β,4,5, 6,7,7α-Hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 5a (484 mg, pale yellow solid), Yield: 97%

Second step

^2-Fluoro-4-methoxyphenyl)-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-4,5,6,7-tetrahydro-1H-pyrazole

Ethyl [3,4- _C ]pyridine-3-carboxylate

Fluoro-4-methoxyphenyl, - Ία-morpholine-fluorene-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-3β,4,5,6,7,7β-six Hydrogen-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 5a (500 mg, 0.86 mmol) was dissolved in 20 mL of tetrahydrofuran, and 2.2 mL of 4 M hydrochloric acid was added at 0 ° C, 0 ° C The reaction was stirred for 2 hours and allowed to stand at 0 ° C for 48 hours. Saturated sodium bicarbonate solution was added dropwise to the reaction mixture ρ Η8, water (30 mL) and dichloromethane (30 mL) were added and the mixture was separated, and the aqueous phase was extracted with dichloromethane (30 mL The residue was washed with a saturated sodium chloride solution (50 mL). Fluoro-4-methoxyphenyl)-7-methyl-6-(4-(3-carbonylmorpholine)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3 , 4-c]pyridine-3-carboxylic acid ethyl ester 5b (150 mg, brownish yellow solid), yield: 34.4%.

third step

1-(2-Fluoro-4-methoxyphenyl)-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-4,5,6,7-tetrahydro-1H-pyridyl Azole

[3,4-c]pyridine-3-carboxamide

1-(2-Fluoro-4-methoxyphenyl)-7-carbonyl-6-(4-(3-carbonylmorpholine)phenyl)-4,5,6,7-tetrahydro-1H- Ethyl pyrazolo[3,4-c]pyridine-3-carboxylate 5b (140 mg, 0.28 mmol) was dissolved in 10 mL of N,N-dimethylformamide, followed by 2.8 mL of formamide, 1 mL Trimethyl formate and 4.2 L of trifluoroacetic acid were stirred at 50 ° C for 30 minutes. 1.2 mL of sodium methoxide C160 mg, 2.76 mmol) in methanol was added and the reaction was stirred at 50 ° C for 5 hours. The reaction mixture was poured into water (20 mL), EtOAc (EtOAc (EtOAc) The residue obtained was purified by silica gel column chromatography eluting eluting with 4,5,6,7-Tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxamide 5 (80 mg, brownish yellow solid), Yield: 60%.

1H NMR (400 MHz, CDC1 ₃ ) δ 7.32-7.41 (m, 5H), 6.78 (s, 1H), 6.71-6.76 (m, 2H), 5.47 (s, 1H), 4.33 (s, 2H), 4.13 (t, 2H), 4.02 (dd, 2H), 3.82 (s, 3H), 3.72 (dd, 2H), 3.39 (t, 2H). Example 6

^4-Methoxyphenyl)-7-carbonyl-6-(4-(2-carbonyl-1,3-oxazin-3-yl)phenyl)-4,5,6,7-tetrahydro- 1H-pyrazole

[3,4-c]pyridine-3-carboxamide

First step

3-(4-iodobenzene)-1,3-oxazin-2-one

Dissolve 1,3-oxazin-2-one 6a (300 mg, 2.97 mmol) and 1,4-diiodobenzene 2b (1.47 g, 4.45 mmol) in 5 mL of dioxane, followed by potassium carbonate ( 1.23 g, 8.91 mmol), iodide (113 mg, 0.59 mmol) and N,N'-dimethylethylenediamine (0.12 g, 1.19 mmol). Filtration, the filtrate was concentrated under reduced pressure, and the obtained residue was purified to silica gel column chromatography to elute to afford the title product 3-(4-iodophenyl)-1,3-oxazin-2-one 6b (527 mg , white solid), Yield: 58.6%.

Second step

3-(4-(3-morpholin-2-carbonyl-5,6-dihydropyridine-1(2H)phenyl)-1,3-oxazin-2-one 3-(4-iodobenzene) -1,3-oxazin-2-one 6b (340 mg, 1.12 mmol) and 3-morpholine-5,6-dihydropyridine-2 (1H ketone 2d (204 mg, 1.12 mmol) dissolved in 6 mL Toluene, potassium phosphate trihydrate (600 mg, 2.24 mmol), iodide (21.40 mg, 0.11 mmol) and trans-N,N"-dimethyl-1,2-cyclohexanediamine ( The reaction was stirred for 5 hours at 110 ° C. The filtrate was concentrated under reduced pressure. Morpholine-2-carbonyl-5,6-dihydropyridine-1(2H-yl)phenyl)-1,3-oxazin-2-one 6c (350 mg, dark green solid yield: 67.3%.

third step

1-(4-Methoxyphenyl)-7α-morpholin-7-carbonyl-6-(4-(2-carbonyl-1,3-oxazin-3-yl)phenyl)-3α,4, 5,6,7,7β-hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester

3-C4-0 morpholine-2-carbonyl-5,6-dihydropyridine-1C2H)-yl)phenyl)-1,3-oxazin-2-one 6c (300 mg, 0.84 mmol) and Z)-2-Chloro-2-(2-(4-methoxyphenyl)hydrazinyl)acetate 2f (216 mg, 0.84 mmol) was dissolved in 8 mL ethyl acetate. 254 mg, 2.52 mmol), and the reaction was stirred at 70 ° C for 60 hours. To the reaction mixture, 20 mL of water was added, and ethyl acetate (10 mL EtOAc) was evaporated. Filtration, filtrate decompression Concentration gave the title product 1 4-methoxyphenyl)-7α-morpholin-7-carbonyl-6 4-P-carbonyl-1,3-oxazin-3-yl)phenyl)-3β,4 , 5,6,7,7β-hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 6d (400 mg, yellow oil), yield: 82.5%.

MS m/z (ESI): 578.4 [M+l]

the fourth step

1-(4-Methoxyphenyl)-7-carbonyl-6-(4-((2-carbonyl-1,3-oxazin-3-yl)phenyl)-4,5,6,7- Ethyl tetrahydro-1H-pyrazolo[3,4- _C ]pyridine-3-carboxylate

-Methoxyphenyl, - Ία-morpholin-7-carbonyl-6 4 2-carbonyl-1,3-oxazin-3-yl)phenyl) -3β,4,5,6,7,7β - Ethyl hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylate 6d (400 mg, 0.69 mmol) was dissolved in 5 mL of tetrahydrofuran, and 1.7 mL of 4 M hydrochloric acid was added at 0 ° C, 0 The reaction was stirred at ° C for 2 hours. The mixture was diluted with aq. The residue was dried over anhydrous sodium sulfate (MgSO4). -(4-((2-carbonyl-1,3-oxazol-3-yl)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine- Ethyl 3-carboxylate 6e (240 mg, yellow oil), Yield: 70.6%.

MS m/z (ESI): 491.4 [M+l]

the fifth step

1-(4-Methoxyphenyl)-7-carbonyl-6-(4-((2-carbonyl-1,3-oxazin-3-yl)phenyl)-4,5,6,7- Tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxamide

1-(4-Methoxyphenyl)-7-yl-6-(4-((2-carbonyl-1,3-oxazin-3-yl)phenyl)-4,5,6, 7-tetrahydrogen

-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 6e (227 mg, 0.46 mmol) was dissolved in 4 mL of N,N-dimethylformamide, followed by the addition of 4.5 mL of formamide. 1 mL of trimethyl orthoformate and 6.8 trifluoroacetic acid were stirred at 50 ° for 30 minutes. 1 mL of a solution of sodium methoxide (250 mg, 2.30 mmol) in methanol was added, and the reaction was stirred at 50 ° C for 5 hours. The reaction mixture was poured into 30 mL of water and extracted with dichloromethane (15 mL EtOAc). The organic phase was combined, washed sequentially with water (50 mL X 2) and saturated sodium chloride solution (50 mL), dried over anhydrous sodium sulfate The filtrate was filtered, and the filtrate was evaporated to dryness. 3-oxazin-3-yl)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxamide 6 C90 mg, pale yellow solid) , Yield: 42%.

1H NMR (400 MHz, CDC1 ₃ ) δ 7.48 (dd, 2H), 7.34 (s, 4H), 6.94 (d, 2H), 6.85 (s, 1H), 5.45 (s, 1H), 4.42 (dd, 2H ), 4.11 (t, 2H), 3.83 (s, 3H), 3.69 (dd, 2H), 3.39 (t, 2H), 2.20 (t 2H). Example 7

1-O-chlorophenyl)-6-C4-C3-fluoro-2-carbonylpyridine-1C2H-phenyl)-7-carbonyl-4,5,6,7-tetrahydro-1H-pyrazole

[3,4-c]pyridine-3-carboxamide

First step

3-fluoro-1-C4-iodobenzene)pyridinium-2C1H)-one

3-Fluoropyridine-2C1H)-one 7a (10 mg, 6.28 mmol, using well-known method literature

"Heterocycles, 28(1), 249-58; 1989" prepared) and 1,4-diiodobenzene 2b (2.50 g, 7.50 mmol) dissolved in 30 mL of dimethyl sulfoxide, followed by the addition of potassium carbonate ( 4.29 g, 31.40 mmol) iodide iodide (0.24 g, 1.26 mmol), stirred at 120 ° C for 12 h. 30 mL of water and 30 mL of dichloromethane were added to the reaction mixture, and the mixture was separated. The aqueous phase was extracted with dichloromethane (30 mL X 5). The organic phase was combined and washed with saturated sodium chloride solution (80 mL) The residue was dried over sodium sulfate, filtered, and then evaporated tolululululululululululululululululululu Ketone 7b (700 mg, pale yellow solid), Yield: 35%.

Second step

3-fluoro-1-(4-(3-morpholin-2-carbonyl-5,6-dihydropyridine-1(2H)phenyl) B-pyridin-2 (1H ketone 3-fluoro-1- (4-iodobenzene) B is pyridine-2 (1H ketone 7b (200 mg, 0.64 mmol) and 3-morpholine-5,6-dihydropyridine-2 (1H ketone 2d (116 mg, 0.64 mmol)) In 8 mL of dioxane, N,N'-dimethylethylenediamine (13 mg, 0.13 mmol) cesium carbonate (620 mg, 1.91 mmol) and iodide (6 mg, 0.03 mmol) were added in that order. The reaction mixture was stirred at 105 ° C for 24 hours. The reaction mixture was cooled to room temperature, filtered, and the filtrate was evaporated evaporated. -(3-morpholin-2-carbonyl-5,6-dihydropyridine-1(2H-yl)phenyl) B-pyridin-2 (1H ketone 7c (80 mg, pale yellow oil), yield: 34.1 %.

third step

1-(3-chlorophenyl)-6-(4-(3-fluoro-2-carbonylpyridine-1(2H-yl)phenyl)-7α-morpholin-7-carbonyl-3β,4,5,6, 7,7β-hexahydropyrazolo[3,4-c]pyridine-3-carboxylate

3-Fluoro-1-(4-(3-morpholin-2-carbonyl-5,6-dihydropyridine-1(2H)phenyl) B-pyridin-2 (1H ketone 7c (80 mg, 0.22) Methyl) and (Z)-2-chloro-2-(2-(3-chlorophenyl)hydrazinyl)acetate 4a (115 mg, 0.44 mmol) Dissolved in 15 mL of ethyl acetate, added triethylamine (222 mg, 2.20 mmol), and stirred at 70 ° C for 3 days. The reaction mixture was combined with EtOAc (EtOAc) (EtOAc) The title is obtained 1-(3-chlorophenyl)-6-(4-(3-fluoro-2-carbonylpyridine-1(2H-yl)phenyl)-7α-morpholine-7-carbonyl-3β,4,5 , 6,7,7β-Hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 7d (135 mg, yellow solid), Yield: 100%.

the fourth step

1-(3-chlorophenyl)-6-(4-(3-fluoro-2-carbonylpyridine-1(2H)-yl)phenyl)-7-carbonyl-4,5,6,7-tetrahydro- 1H-pyrazole

Ethyl [3,4- _C ]pyridine-3-carboxylate

1-(3-Chlorophenyl)-6-(4-(3-fluoro-2-carbonylpyridine-1(2H)-yl)phenyl, -Ία-morpholine-indole-carbonyl-3β,4,5 ,6,7,7β-hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 7d (135 mg, 0.23 mmol) dissolved in 10 mL of tetrahydrofuran, 0 ° C added 0.3 mL 4 M hydrochloric acid, stirring at 0 ° C for 4 hours, at 0 ° C for 12 hours, stirring to room temperature and stirring for 2 hours. Add saturated sodium bicarbonate solution to the reaction solution ρ Η 8 with ethyl acetate (20 mL X 3) Extraction, the organic phase was combined, washed with a saturated sodium chloride solution (30 mL), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated to give the title product 1-(3-chlorobenzene)-6-(4 -(3-fluoro-2-carbonylpyridine-1(2H)phenyl)-7-carbonyl-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3 Ethyl formate 7e (89 mg, yellow solid), Yield: 77%.

the fifth step

[3,4-c]pyridine-3-carboxamide

1-0-chlorobenzene)-6-(4-(3-fluoro-2-carbonylpyridine-1(2H)-yl)phenyl)-7-carbonyl-4,5,6,7-tetrahydro-1H - Pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 7e (89 mg, 0.18 mmol) was dissolved in 8 mL of N,N-dimethylformamide, followed by 1 mL of formamide, 1 mL Trimethyl orthoformate and 3 L of trifluoroacetic acid were stirred and reacted at 50 ° C for 30 minutes. 5 mL of sodium methoxide C 48 mg, 0.88 mmol) in methanol was added, and the reaction was stirred at 50 ° C for 2 hours. The reaction mixture was poured into water (10 mL), EtOAc (EtOAc m. The residue obtained was purified by HPLC column to give the title product 1-(3-chlorobenzene)-6-(4-(3-fluoro-2-carbonylpyridine-1(2Hyl)phenyl)-7-carbonyl- 4,5,6,7-Tetrahydro-1H-pyrazolo[3,4- _C ]pyridine-3-carboxamide 7 (12 mg, white solid), yield: 14.4%.

1H NMR (400 MHz, CDC1 ₃ ) δ 7.61 (s, 1H), 7.36-7.52 (m, 7H), 7.14-7.20 (m, 2H), 6.84 (s, 1H), 6.16-6.21 (m, 1H) , 5.48 (s, 1H), 4.19 (t, 2H), 3.43 (t, 2H). Example 8

1- 2,3-Dihydrobenzofuran-5-yl)-7-carbonyl-6-C4-C2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro- 1H-pyrazolo[3,4-c]pyridine-3-carboxamide

First step

(Z)-2-Chloro-2-(2-(2,3-dihydrobenzofuran-5-yl)arylene)ethyl acetate 2,3-dihydrobenzofuran-5-amine 8a (2.20 g, 16.30 mmol, patent application

"WO2003049702" prepared by the method disclosed) added to 7 mL of water, slowly added dropwise 4 mL of concentrated hydrochloric acid and 2.8 mL of 40% aqueous sodium nitrite solution at 0 ° C, stirred at 0 ° C for 1 hour, stored as a reserve 1. Ethyl 2-chloroacetoacetate (2.90 g, 17.80 mmol) and sodium acetate (2.70 g, 32.60 mmol) were added to a mixture of 13 mL of acetone and water (V/V = 6:7), and added dropwise at 0 °C. The stock solution 1 was added dropwise, and the reaction was stirred at 0 ° C for 1 hour and at 0 ° C for 30 minutes. The liquid was separated, 4 mL of methanol was added to the organic phase, and the reaction was stirred at 0 ° C for 5 hours, filtered, and the filter cake was dried to give the title product (Z)-2-chloro-2-(2-(2,3-dihydro) Ethyl benzofuran-5-yl)indenyl)acetate 8b (1.20 g, yellow solid), yield: 28%.

Second step

1-(2,3-dihydrobenzofuran-5-yl 7α-morpholine-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)benzene

-3β,4,5,6,7,7β-hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester (Z)-2-chloro-2-(2 -(2,3-dihydrobenzofuran-5-yl)hydrazino)ethyl acetate 8b (402 mg, 1.50 mmol) and 3-morpholine-1-(4-(2-carbonylpiperidine-1) -yl)phenyl)-5,6-dihydropyridine-2 (1H ketone 3c (355 mg, 1 mmol) was dissolved in 20 mL ethyl acetate, triethylamine (505 mg, 5 mmol), 68 ° C. The reaction was stirred for 3 days. The reaction mixture was cooled to room temperature, then added with 20 mL of water, and extracted with ethyl acetate (20 mL of EtOAc). The organic phase was combined and washed with saturated sodium chloride (20 mL) Drying, filtration and concentration under reduced pressure afforded the title product 1-(2,3-dihydrobenzofuran-5-yl)-7α-morpholin-7-carbonyl-6 4-(2-carbonylpiperidine-1 -yl)phenyl) -3β,4,5,6,7,7β-hexahydro-1H-borazolo[3,4-c]ethylidene-3-carboxylate 8c (630 mg, black Solid), Yield: 100%. third step

1-(2,3-dihydrobenzofuran-5-yl)-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7- Tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylate 1-(2,3-Dihydrobenzofuran-5-yl)-7α-morpholin-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-3β, 4,5,6,7,7β-hexahydro-1H-borazolo[3,4-c]ethylidene-3-carboxylate 8c (630 mg, 1 mmol) was dissolved in 20 mL of tetrahydrofuran. 1.25 mL of 4 M hydrochloric acid was added dropwise at 0 ° C, and the reaction was stirred at 0 ° C for 2 hours. The saturated sodium bicarbonate solution was added dropwise to a mixture of EtOAc (EtOAc) (EtOAc) (EtOAc (EtOAc) filtered, concentrated under reduced pressure, purified by silica gel column chromatography in a developing solvent A purification system resulting residue, to give the title product 1- (2,3-dihydro-benzofuran-5-yl) - ₇ - carbonyl - ₆ - ( ; 4-(; 2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 8d (330 mg, colorless oily yield: 66%.

the fourth step

1-(2,3-dihydrobenzofuran-5-yl)-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7- Tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxamide

1-(2,3-Dihydrobenzofuran-5-yl)-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7 -tetrahydro-1H-pyrazolo[3,4-c|pyridine-3-carboxylic acid ethyl ester 8d (330 mg, 0.66 mmol) was dissolved in 20 mL of N,N-dimethylformamide, followed by 2 mL Formamide, 2 mL of trimethyl orthoformate and 9 trifluoroacetic acid were stirred at 50 ° C for 30 minutes. 10 mL of sodium methoxide (178 mg, 3.30 mmol) in methanol was added and the reaction was stirred at 50 ° C for 2 hours. The reaction solution was poured into 20 mL of water, and extracted with dichloromethane (30 mL EtOAc). The organic phase was combined, washed sequentially with water (20 mL X 2) and saturated sodium chloride solution (20 mL), dried over anhydrous sodium sulfate After filtration, the filtrate was concentrated under reduced pressure, and the residue obtained was purified by HPLC to afford the title product 1-(2,3-dihydrobenzofuran-5-yl)-7-carbonyl-6-(4-(2) -carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4- _C ]pyridine-3-carboxamide 8 (120 mg, yellow-brown solid ), Yield: 38.6%.

1H NMR (400 MHz, CDC1 ₃ ) δ 7.36 (s, 3H), 7.25-7.28 (m, 2H), 6.82-6.90 (m, 3H), 5.44 (s, 1H), 4.62 (t, 2H), 4.12 (t, 2H), 3.59-3.65 (m, 2H), 3.39 (t, 2H), 3.25 (t, 2H), 2.55-2.65 (m, 2H), 1.91-2.00 (m, 4H). Example 9

i - (difluoromethoxy)phenyl)-7-carbonyl-6-(4-P-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro-1H-pyridyl Zoxa[3,4-c]pyridine-3-carboxamide

First step

(Z)-2-Chloro-2-(2-(4-(difluoromethoxy)phenyl)phosphonium)acetate ethyl 2,3-dihydrobenzofuran-5-amine 9a (4 g, 25.10 mmol, purchased from "Shanghai Yuhong Chemical Technology Co., Ltd.") added to 16 mL of water, slowly added dropwise 6.1 mL of concentrated hydrochloric acid and 20 mL of sodium nitrite (2.10 g, 30.10 mmol) in 0 °C, The reaction was stirred at 0 ° C for 1.5 hours and stored as a stock solution 1. Ethyl 2-chloroacetoacetate (4.10 g, 25.10 mmol) and sodium acetate (4.74 g, 57.80 mmol) were added to a mixed solution of 72 mL of acetone and water (V/V = 4:5) at 0 °C. The stock solution 1 was added dropwise, and the reaction was stirred at 0 ° C for 3 hours. 150 mL of methylene chloride was added to the reaction mixture, and the aqueous layer was extracted with dichloromethane (150 mL EtOAc). The organic phase was combined and washed with water (150 mL) and saturated sodium chloride (150 mL). Drying over anhydrous sodium sulfate, filtration, and EtOAcjjjjjjjjj 9b (5.70 g, yellow solid), Yield: 78%.

MS m/z (ESI): 291.1 [M-l]

Second step

L-(4-(;Difluoromethoxy)phenyl)-7α-morpholine-7-carbonyl-6-(4-0carbonylpiperidin-1-yl)phenyl)-3α,4,5, 6,7,7β-hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester

(Z)-2-Chloro-2-(2-(4-(difluoromethoxy)phenyl)indenyl)acetate 9b (256 mg, 0.87 mmol) and 3-morpholine-1 -(4-(2-Carbopiperidin-1-yl)phenyl)-5,6-dihydropyridine-2 (1H ketone 3c (310 mg, 0.87 mmol) was dissolved in 5 mL ethyl acetate. Ethylamine (264 mg, 2.61 mmol) was stirred at 80 ° C for 18 hours. The reaction mixture was cooled to room temperature, 15 mL water was added, and the mixture was separated, and the aqueous phase was extracted with ethyl acetate (20 mL X 3 ). Washed with a saturated aqueous solution of sodium chloride (20 mL), dried over anhydrous sodium sulfate 7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-3β,4,5,6,7,7β-hexahydro-1H-pyrazolo[3,4-c Pyridine-3-carboxylic acid ethyl ester 9c (370 mg, brown oil), yield: 69%.

MS m/z (ESI): 612.3 [M+ 1]

third step

i - (difluoromethoxy)phenyl)-7-carbonyl-6-(4-P-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro-1H-pyridyl Ethazo[3,4- _C ]pyridine-3-carboxylate

1-(4-(Difluoromethoxy)phenyl yia-morpholine-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)benzene -3β,4,5,6,7,7β-hexahydro-1H-pyrazolo[3,4-c|pyridine-3-carboxylic acid ethyl ester 9c (370 mg, 0.60 mmol) dissolved in 15 mL of tetrahydrofuran In the mixture, 0.5 mL of 4 M hydrochloric acid was added dropwise at 0 ° C, the reaction was stirred at 0 ° C for 8 hours, and allowed to stand at 0 ° C for three days. The saturated sodium bicarbonate solution was added dropwise to a mixture of EtOAc (EtOAc) (EtOAc) (EtOAc (EtOAc) Concentration under reduced pressure gave 1-(4-(difluoromethoxy)phenyl)-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)- Ethyl 5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylate 9d (360 mg, brownish yellow solid). Yield: 100%.

the fourth step

(difluoromethoxy)phenyl)-7-carbonyl-6-(4-P-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro-1H-pyrazole [3,4-c]pyridine-3-carboxamide

1-(4-(Difluoromethoxy)phenyl)-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetra Hydrogen-1H-pyrazolo[3,4-c|pyridine-3-carboxylic acid ethyl ester 9d (317 mg, 0.60 mmol) was dissolved in 9 mL of N,N-dimethylformamide, followed by 2 mL of formamide , l mL of trimethyl orthoformate and 3 L of trifluoroacetic acid, and stirred at 50 ° C for 30 minutes. 10 mL of sodium methoxide (163 mg, 3.02 mmol) in methanol was added and the reaction was stirred at 50 ° C for 1.5 hours. The reaction mixture was poured into 20 mL of water, and extracted with methylene chloride (20 mL EtOAc). The organic phase was washed with water (20 mL) and saturated sodium chloride (20 mL) the filtrate was concentrated under reduced pressure and the resulting residue was purified by HPLC preparative column, to give the title product 1- (4- (difluoromethoxy) phenyl) -7-carbonyl - ₆ - (4- (2-oxo-piperidine -1,5,6,7-tetrahydro-1H-pyrazolo[3,4- _C ]pyridine-3-carboxamide 9 (120 mg, pale yellow solid), yield : 40.1%.

1H NMR (400 MHz, DMS0-d6) δ 7.75 (s, 1H), 7.66 (d, 2H), 7.48 (s, 1H), 7.35 (d, 2H), 7.31 (s, 1H), 7.25-7.30 ( m, 4H), 4.06 (t, 2H), 3.59 (t, 2H), 3.20 (t, 2H), 2.38 (t, 2H), 1.81-1.89 (m, 4H). Example 10

(3-fluoro-4-methoxyphenyl)-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro-1H -pyrazole

[3,4-c]pyridine-3-carboxamide

First step

(Z)-2-Chloro-2-(2-(3-fluoro-4-methoxyphenyl)phosphonium)acetate ethyl 3-fluoro-4-methoxyaniline 10a (1.40 g, 10 Methyl) was added to 10 mL of water. At 0 ° C, 2.5 mL of concentrated hydrochloric acid and 1.8 mL of 40% aqueous sodium nitrite were slowly added dropwise, and the reaction was stirred at 0 ° C for 1.5 hours, and stored as a reserve liquid 1. Ethyl 2-chloroacetoacetate (1.89 g, 10.90 mmol) and sodium acetate (1.60 g, 20 mmol) were added to a mixed solution of 10 mL of acetone and water (V/V = 2:3) at 0 °C. The stock solution 1 was added dropwise, and the reaction was stirred at 0 ° C for 1.5 hours and at 0 ° C for 12 hours. 50 mL of methylene chloride was added to the reaction mixture, and the aqueous layer was extracted with dichloromethane (50 mL×2). The organic phase was combined and washed with water (50 mL) and saturated sodium chloride (50 mL). The residue was dried over anhydrous sodium sulfate, filtered and evaporated. 10b (2.20 g, black solid), Yield: 80.3%.

Second step

1-(3-Fluoro-4-methoxyphenyl)-7β-morpholine-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-3β,4,5 ,6,7,7α-hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester

(Z)-2-Chloro-2-(2-(3-fluoro-4-methoxyphenyl)hydrazinyl)acetate 10b (178 mg, 0.50 mmol) and 3-morpholin-1- (4-(2-Carbopiperidin-1-yl)phenyl)-5,6-dihydropyridine-2 (1H ketone 3c (274 mg, 1 mmol) dissolved in 20 mL of ethyl acetate Amine (1.01 g, 10 mmol), stirred at 80 ° C for 3 hours, and stirred at 70 ° C for 12 hours. The reaction solution was cooled to room temperature, 20 mL water was added, and the mixture was separated. 3) The organic phase was combined, washed with EtOAc EtOAc (EtOAc) Morpholine-7-carbonyl-6-^2-carbonylpiperidin-1-yl)phenyl)-3β,4,5,6,7,7α-hexahydro-1H-pyrazolo[3,4-c Pyridine-3-carboxylic acid ethyl ester 10c (281 mg, black solid), Yield: 94.6%.

third step

Ethyl [3,4- _C ]pyridine-3-carboxylate

1-(3-Fluoro-4-methoxyphenyl)-7α-morpholin-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-3β,4, 5,6,7,7β-hexahydro-1H-indolezolo[3,4-c]indolepyridin-3-carboxylate 10c (281 mg, 0.47 mmol) dissolved in 20 mL of tetrahydrofuran, 0° 1.2 mL of 4 M hydrochloric acid was added dropwise to C, and the reaction was stirred at 0 ° C for 3 hours, and allowed to stand at 0 ° C for three days. Saturated sodium bicarbonate solution was added dropwise until the reaction mixture was EtOAc (EtOAc) The combined organic phases were washed with EtOAc EtOAc EtOAc (HHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHHH Carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid Ethyl ester 10d (240 mg, colorless oil), Yield: 100%.

the fourth step

[3,4-c]pyridine-3-carboxamide

_1- (3-Fluoro-4-methoxyphenyl)-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-4,5,6,7-tetra Hydrogen-1 pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 10d (230 mg, 0.45 mmol) was dissolved in 6 mL of N,N-dimethylformamide, followed by 2 mL of formamide. l mL of trimethyl orthoformate and 3 L of trifluoroacetic acid were stirred at 50 ° C for 30 minutes. 10 mL of sodium methoxide (122 mg, 2.27 mmol) in methanol was added and the mixture was stirred at 50 ° C for 1.5 hours. The reaction mixture was poured into 20 mL of water and extracted with dichloromethane (15 mL X 4). The organic phase was combined, washed sequentially with water (20 mL) and saturated sodium chloride (20 mL) the filtrate was concentrated under reduced pressure and the resulting residue was purified by preparative HPLC column to give the title product 1- (3-fluoro-4-methoxyphenyl) -7-carbonyl - ₆ - (4- (2-oxo-piperidine -1-yl)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4- _C ]pyridine-3-carboxamide 10 (80 mg, yellow solid), yield: 37%.

1H NMR (400 MHz, DMSO-d6) δ 7.75 (s, 1H), 7.57 (d, 1H), 7.46 (s, 1H), 7.42 (dd, 1H) _: 7.35 (d, 2H), 7.27 (d, 2H), 7.26 (t, 1H), 4.04 (t, 2H), 3.88 (s, 3H), 3.59 (t, 2H), 3.19 (t, 2H), 2.38 (t, 2H), 1.79-1.90 (m , 4H). Example 11

₇ -carbonyl- _6- (4-(2-carbonylpiperidin-1-yl)phenyl)-l-(4-(trifluoromethoxy)phenyl)-4,5,6,7-tetrahydro -1H-pyrazolo[3,4-c]pyridine-3-carboxamide

7 _β -morpholine-7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-1 -(4-(trifluoromethoxy)phenyl)-3α,4, 5,6,7,7β-hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester

(Z)-2-Chloro-2-(2-(4-(trifluoromethoxy)phenyl)indenyl)acetate 11a (279 mg, 0.90 mmol, using a known method " Bioorganic & Medicinal Chemistry Letters, 2010, 20(15), 4683-4688 "Prepared" and 3-morpholine-1-(4-(2-carbonylpiperidin-1-yl)phenyl)-5,6-dihydro Pyridine-2 (1H ketone 3c (160 mg, 0.45 mmol) was dissolved in 5 mL ethyl acetate, triethylamine (136 mg, 1.35 mmol) was added, and the reaction was stirred at 80 ° C for 65 hours. Wash with water (5 mL) and EtOAc (EtOAc (EtOAc) Carbonylpiperidin-1-yl)phenyl)sodium(4-(trifluoromethoxy)phenyl)-3β,4,5,6,7,7α-hexahydro-1H-pyrazolo[3,4 -c] pyridine-3-carboxylate llb (210 mg, brown oil), yield: 74.2%.

Second step

₇ -carbonyl- _6- (4-(2-carbonylpiperidin-1-yl)phenyl)-1-(4-(trifluoromethoxy)phenyl)-4,5,6,7-tetrahydro -1H-pyrazolo[3,4- _C ]pyridine-3-carboxylate

Ία-morpholine-Ί-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-1 -(4-(trifluoromethoxy)phenyl)-3β,4, 5,6,7,7β-hexahydro-1H-indolozolo[3,4-c]indolepyridin-3-carboxylate lib (210 mg, 0.33 mmol) dissolved in 15 mL of tetrahydrofuran, 0° 0.3 mL of 4 M hydrochloric acid was added dropwise under C, and the reaction was stirred at 0 ° C for 3 days. The saturated sodium bicarbonate solution was added dropwise to a mixture of EtOAc (EtOAc) (EtOAc (EtOAc) (EtOAc) Concentration under reduced pressure gave the title product 7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-1-(4-(trifluoromethoxy)phenyl)-4. 5,6,7-Tetrahydro-1H-cyclopyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester, mp C240 mg, mp.

third step

₇ -carbonyl- _6- (4-(2-carbonylpiperidin-1-yl)phenyl)-1-(4-(trifluoromethoxy)phenyl)-4,5,6,7-tetrahydro -1H-pyrazolo[3,4-c]pyridine-3-carboxamide

7-Carbon-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-1-(4-(trifluoromethoxy)phenyl)-4,5,6,7-tetra Hydrogen-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 11c (181 mg, 0.33 mmol) was dissolved in 3 mL of N,N-dimethylformamide, followed by 2 mL of formamide , l mL of trimethyl orthoformate and 3 L of trifluoroacetic acid, and stirred at 50 ° C for 30 minutes. A 10 mL solution of sodium methoxide (90 mg, 1.67 mmol) in methanol was added, and the reaction was stirred at 50 ° C for 1.5 hours. The reaction mixture was poured into 20 mL of water and extracted with dichloromethane (15 mL X 3). The organic phase was combined, washed sequentially with water (20 mL) and saturated sodium chloride (20 mL) The filtrate was concentrated under reduced pressure, and the obtained residue was purified to purified crystals crystals to afford the title product 7-carbonyl-6-(4-(2-carbonylpiperidin-1-yl)phenyl)-1-(4-(3) Fluoromethoxy)phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxamide ll (200 mg, yellow solid), yield: 100%.

1H NMR (400 MHz, DMS0-d6) δ 7.75-7.78 (m, 3H), 7.47-7.51 (m, 3H), 7.36 (d, 2H), 7.28 (d, 2H), 4.06 (t, 2H), 3.59 (t, 2H), 3.21 (t, 2H), 2.38 (t, 2H), 1.78-1.90 (m, 4H). Example 12 L-(3-Chlorophenyl)-7-carbonyl-6-(4-(2-carbonyl-1,3-oxazin-3-yl)phenyl)-4,5,6,7-tetrahydro- 1H-pyrazole

First step

1-(3-Chlorophenyl)-7α-morpholin-7-carbonyl-6-(4-(2-carbonyl-1,3-oxazin-3-yl)phenyl)-3a,4,5, 6,7,7a-hexahydrogen

-1H-pyrazolo[3,4-c]pyridine-3-carboxylate

3-C4-0 morpholine-2-carbonyl-5,6-dihydropyridine-1C2H)-yl)phenyl)-1,3-oxazin-2-one 6c (346 mg, 0.97 mmol) and Z)-2-Chloro-2-(2-(3-chlorophenyl)hydrazinyl)acetate 4a (505 mg, 1.94 mmol) was dissolved in 20 mL ethyl acetate and triethylamine (0.7 mL) , 4.85 mmol), stir the reaction at 70 °C for 3 days. To the reaction mixture, 40 mL of water, EtOAc (EtOAc m. The title product 1-(3-chlorophenyl) _{-7βmorpholin} -7-carbonyl-6-(4-(2-carbonyl-1,3-1,3-oxazin-3-yl)phenyl)-3a,4, 5,6,7,7a-Hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylate 12a (770 mg, yellow oil), Yield: 100%.

MS m/z (ESI): 582.2 [M+l]

Second step

L-(3-Chlorophenyl)-7-carbonyl-6-(4-(2-carbonyl-1,3-oxazin-3-yl)phenyl)-4,5,6,7-tetrahydro- 1H-pyrazole

Ethyl [3,4- _C ]pyridine-3-carboxylate

1-0 chlorophenyl)-7α-morpholin-7-carbonyl-6-(4-(2-carbonyl-1,3-oxazin-3-yl)phenyl)-3a,4,5,6 ,7,7a-Hexahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 12a (563 mg, 0.97 mmol) was dissolved in 6 mL of tetrahydrofuran, 2.4 mL 4 M hydrochloric acid, 0 The reaction was stirred at ° C for 2.5 hours, and the reaction was stirred at room temperature for 3 hours. The saturated sodium bicarbonate solution was added dropwise until the reaction mixture was 77, extracted with dichloromethane (10 mL×2), and the organic phase was combined, washed sequentially with water (15 mL) and saturated sodium chloride solution (50 mL). The residue was purified by EtOAcjjjjjjjjjjjjjj (2-carbonyl-1,3-oxazol-3-yl)phenyl)- 4,5,6,7-tetrahydro-1H-cyclopyrazolo[3,4-c]pyridin-3- Ethyl formate 12b (366 mg, yellow solid), Yield: 76.4%. MS m/z (ESI): 512.2 [M+18]

third step

[3,4-c]pyridine-3-carboxamide

1-(3-Chlorophenyl)-7-methyl-6-(4-(2-carbonyl-1,3-oxazin-3-yl)phenyl)-4,5,6,7-tetra Hydrogen-1H-port bisazolo[3,4-c]pyridine-3-carboxylic acid ethyl ester 12b (227 mg, 0.46 mmol) was dissolved in 3 mL of N,N-dimethylformamide, followed by 4.5 mL. The amide, 1 mL of trimethyl orthoformate and 6.8 L of trifluoroacetic acid were stirred at 50 ° C for 30 minutes. 0.5 mL of sodium methoxide C248 mg, 2.29 mmol) in methanol was added and the reaction was stirred at 50 ° C for 1 hour. Add 30 mL of water, extract with methylene chloride (15 mL of EtOAc), EtOAc (EtOAc)EtOAc. Concentration, the obtained residue was purified to purified crystals eluted eluted eluted eluted Phenyl)-4,5,6,7-tetrahydro-1H-pyrazolo[3,4-c]pyridine-3-carboxamide 12 (100 mg, white solid), yield: 46.8%.

MS m/z (ESI): 483.3 [M+18]

1H NMR (400 MHz, CDC1 ₃ ) δ 7.60 (s, 1H), 7.46-7.53 (m, 1H), 7.33-7.44 (m, 6H), 6.91 (s, 1H), 5.92 (s, 1H), 4.40 -4.48 (m, 2H), 4.13 (t, 2H), 3.71 (t, 2H), 3.40 (t, 2H), 2.18-2.26 (m, 2H). Test example:

Biological evaluation

The invention is further described below in conjunction with the test examples, but these examples are not intended to limit the scope of the invention.

The experimental methods in which the specific conditions are not specified in the test examples of the present invention are usually carried out according to conventional conditions or according to the conditions recommended by the manufacturer of the product. Reagents not specified for specific sources, are routine reagents purchased by the market. Test Example 1 Fluorescence detection of biological activity of Factor Xa inhibitor

Experimental material

Enzyme: Factor Xa Protease (Thermo 32521),

Substrate: Factor Xa specific substrate (Chromogenix, S-2222TM),

Buffer: 50 mM Tris pH 8.3, 200 mM NaCl, 5 mM CaCl ₂ .

2. Experimental steps

20 mM test compound dissolved in 100% DMSO was diluted with buffer to 100 nM, 33.3 nM, 11.1 nM, 3.7 nM, 1.23 nM, 0.41 nM, 0.14 nM, 0.05 nM. At the same time, dilute the 2 mM substrate to l.lmM with buffer, and dilute the 25 μL enzyme storage solution to 25 Μ with buffer.

In the test, 84 μM of the prepared substrate solution was added to a 96-well plate. At the same time, 10 μΙ gradient compound was added to each well (10 μΙ 2.5% DMSO was added to the blank control well); 6 μΙ 25ηΜ enzyme solution. After incubating for 1 hour at room temperature after shaking, the absorbance was measured at 405 nm. The absorbance values were curve fitted with Prism and the IC50 was obtained. Compound number IC ₅₀ (Factor Xa) / (nM)

Example 1 0.48

Example 2 0.97

Example 3 1.35

Example 4 1.53

Example 5 0.30

Example 6 1.22

Example 7 0.52

Example 8 0.76

Example 9 1.65

Example 10 0.54

The compounds of the invention have significant inhibitory activity against Factor Xa. Test Example 2 Determination of anticoagulant effect in human blood in vitro

Blood collection: Intravenous blood collection, using a disposable plastic syringe containing 3.8% sodium citrate (1:9 volume, one part of citric acid and nine parts of blood anticoagulation). The venous blood was centrifuged at 2000 g for 10 minutes (4 ° C), and the upper platelet-poor plasma was taken for subsequent testing.

Detection of Factor Xa activity in blood: At 37 ° C, in 20 ul of platelet-poor human plasma, add 0.4 ul of compound/DMSO, add 20 ul of luminescent substrate (1.75 mM), 20 ul RVV (0.7 mU/ml, Mixed with 100 mM CaCb 1 :1). COD 405 nm) was continuously detected by a microplate reader for 10 min to evaluate the inhibitory effect of the drug on blood agglutination.

The compounds of the invention have significant inhibitory activity against human blood agglutination. Pharmacokinetic evaluation

Test Example 3. Pharmacokinetic test of the compound of the present invention

1, abstract

Rats were used as test animals, and the concentration of the drug in plasma at different times after administration of the compounds of Examples 1, 9 and 10 by intragastric administration was determined by LC/MS/MS. The pharmacokinetic behavior of the compounds of the present invention in rats was investigated and their pharmacokinetic characteristics were evaluated.

2. Test plan

2.1 Test drugs

The compound of Example 1, the compound of Example 9, and the compound of Example 10. 2.2 Test animals

Healthy adult SD rats, 12 males and females, divided into 3 groups, 4 in each group, purchased from Shanghai Xipuer-Beikai Experimental Animal Co., Ltd., animal production license number: SCXK (Shanghai) 2008-0016.

2.3 Drug preparation

Weigh the appropriate amount, add 25 μM Tween 80, add Labrasol to the final volume, and make a 0.6 mg/ml solution by sonication.

2.4 Administration

Twelve SD rats, male and female, were divided into 3 groups. After fasting overnight, they were intragastrically administered at a dose of 3.0 mg/kg and a dose of 5 ml/kg.

3, operation

0.1 ml of blood was collected before, and 0.5, 1, 2, 4, 6, 8, 11, 24 hours after administration, placed in an EDTA anticoagulation tube, centrifuged at 3500 rpm for 5 min, and plasma was separated at -20 °C. save. Eat 2 hours after administration.

The content of the test compound in the plasma of rats after intragastric administration of different compounds was determined by LC/MS/MS method. The linear range of the analytical method was 5.00-2000 ng/ml and 1.00-500 ng/ml, respectively, and the lower limit of quantification was 5.00 ng/ml and 1.00 ng/ml, respectively; plasma samples were pretreated with precipitated protein for analysis.

4, pharmacokinetic parameters results

The pharmacokinetic parameters of the compounds of the invention are as follows:

Conclusion: The compound of the present invention has good absorption of the drug and has obvious pharmacological absorption effect. Human pharmacokinetics and coagulation parameter test (PK/PD) test

Test Example 4. Inventive Example 1 Human body pharmacokinetics and coagulation parameter detection (PK/PD) test of APIXABAN

Dosage: 5mg / person / day

Number of people: 4 people, male healthy volunteers, no previous history of heart disease

Mode: Oral, single dose

Time of blood collection: before administration (Oh) and after administration 0.5, 1.5, 3, 6, 10, 24, 48h

Blood volume: Pharmacokinetics: 3.0 ml, whole blood was collected at a specified time point using a vacuum heparin anticoagulation tube, centrifuged at 3000 rpm for 4 min at 10 °C, and the plasma was collected at -20 °C, and the blood drug concentration was measured.

Efficacy: 2.7 ml, 3 ml of blood was collected by citrate anticoagulation tube, centrifuged at 3000 rpm for 4 min for 10 min, and absorbed. Plasma was tested for coagulation parameters.

Detailed prescription of the drug:

4 healthy volunteers, fasted for 8 hours or more overnight, free of breakfast, orally administered to the test tablets (the compound of Example 1 and the conventional excipients in the field were mixed with dry compressed tablets, the specific prescriptions are shown in the following table, APIXABAN use Commercially available tablets), the subjects were given Chinese food after 4 hours of administration, and given for 10 hours after administration. Before administration (Oh) and 0.5, 1.5, 3 6 10 24 48h after administration, 3 ml of venous blood was taken, and whole blood was collected at a prescribed time point using a vacuum heparin anticoagulation tube, and then centrifuged at 4 degrees (3000 rpm) for 10 minutes. Plasma was obtained, and the obtained plasma was dispensed into two tubes, stored at -20 ° C, and sent to Shanghai Hengrui for testing.

Pharmacodynamic parameters test protocol: 4 healthy volunteers, fasting for more than 8 hours overnight, free breakfast, oral administration of the test tablets on a fasting basis, subjects given Chinese food after 4 hours of administration, given 10 hours after administration . Before administration (Oh) and 0.5, 1.5 3 6 10, 24 48 h after administration, 2.7 ml of venous blood was taken, and whole blood was collected at a prescribed time point using a vacuum heparin anticoagulation tube, and then centrifuged at 4 degrees (3000 rpm) for 10 minutes. Plasma, blood coagulation parameters were tested (the present invention tested the activated partial prothrombin time and prothrombin time, ie aPTT and PT).

Note: Two-cycle cross-administration, the interval between two drug administrations is 96h; the PK/PD data obtained from the test are as follows:

PK parameters:

Parameters Example 1 APIXABAN

1.5 2.25

C (ng/ml) 87.175 250.75

AUC o (ng/ml*h) 850.2225 2655.562

AUC o (ng/ml*h) 914.9553 2681.848 ti/ ₂ (h) 7.830894 7.147677

MRT Q-CO (h) 10.68394 9.67734 PD parameters (APTT):

PD parameters (PT):

According to the combined PK and PD data, the compound of the present invention has a drug efficacy lower than that of apixaban, and the in vivo exposure is less than apixaban, and the obtained drug effect is still much better than apixaban. It can be seen that the compound of the present invention obtains the same drug. At the same time, the required blood concentration and exposure are greatly reduced, thus greatly reducing toxicity and adverse reactions.

Claims

Claims:

1. A compound as shown below or a tautomer, a mesogen, a racemate, an enantiomer,

2. A compound as shown below or a tautomer, a mesogen, a racemate, an enantiomer, a diastereomer, a mixture thereof, and a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable salt thereof :

4. A pharmaceutical composition comprising a therapeutically effective amount of a compound according to claim 1 or a tautomer thereof, a mesophil, a racemate, an enantiomer, Diastereomers, mixtures thereof, and pharmaceutically acceptable salts thereof, and pharmaceutically acceptable carriers, diluents or excipients.

5. A compound according to claim 1 or a tautomer, a mesogen, a racemate, an enantiomer, a diastereomer, a mixture thereof, and a pharmaceutically acceptable drug thereof Use of a salt, or a pharmaceutical composition according to claim 4, for the preparation of a prophylactic and/or therapeutic thromboembolic disease.

6. The use according to claim 5, wherein the disease is selected from the group consisting of a cardiac infarction, angina pectoris, angioplasty or aortic coronary artery shunt re-occlusion and restenosis, stroke, transient ischemic attack, Peripheral arterial occlusive disease, pulmonary embolism or deep vein thrombosis.

7. A compound according to claim 1 or a tautomer, a mesogen, a racemate, an enantiomer, a diastereomer, a mixture thereof, and a pharmaceutically acceptable drug thereof Use of a salt, or a pharmaceutical composition according to claim 4, for the preparation of a medicament for preventing and/or treating a disease which is positively affected by inhibition of factor Xa Use in.

8. A compound according to claim 1 or a tautomer, a mesogen, a racemate, an enantiomer, a diastereomer, a mixture thereof, and a pharmaceutically acceptable drug thereof Use of a salt, or a pharmaceutical composition according to claim 4, for the manufacture of a medicament for the treatment of disseminated intravascular coagulation.

9. A compound according to claim 1 or a tautomer, a mesogen, a racemate, an enantiomer, a diastereomer, a mixture thereof, and a pharmaceutically acceptable drug thereof Use of a salt, or a pharmaceutical composition according to claim 4, for the manufacture of a medicament for inhibiting Factor Xa.