CN1260215C - Antagon of endostadin receptor pyrazole carboxylic acids - Google Patents

Abstract

The present invention relates to a compound of a general formula (I) and salts thereof, wherein R1, R2, and R3 are organic substituent groups. The compound of the present invention has the antagonistic activity of endothelin receptor, and can be used for treating cardio-cerebrovascular diseases, tumours, diabetes, kidney diseases, asthma, etc.

Description

Pyrazole carboxylic acid endothelin receptor antagonist

technical field

The present invention relates to novel pyrazole carboxylic acid endothelin receptor antagonists and their application in treating cardiovascular and cerebrovascular diseases, tumors, diabetes, nephropathy and asthma.

Background technique

The mortality rate of cardiovascular and cerebrovascular diseases ranks first among various diseases, and the final cause of death is overwhelmingly cardiac hypertrophy, heart failure (heart failure), stroke and fatal arrhythmia. Especially in recent years, the fatality rate of severe heart failure has remained high, which has become a difficult problem in the international medical community. These diseases currently lack effective therapeutic drugs.

Endothelin (ET) is one of the strongest known vasoconstrictor factors discovered in 1988, and ETs include ET-1, ET-2 and ET-3. There are two main types of ET receptors in mammals: _ETA and _ETB . The ET-1 selective _ETA receptor mainly expressed in the vessel wall and smooth muscle cells mediates the vasoconstriction and mitogenic effect of ET-1; the ET _B receptor has the effect on ET-1, ET-2 and ET -3 is a non-selective, _ETB receptor expressed on endothelial cells that mediates the release of nitric oxide (NO) and prostacyclin _I2 ( _PGI2 ) causing vasodilation. Expressed in smooth muscle cells, it mediates vasoconstriction after _ETB receptor activation.

ET is widely involved in the pathological process of various diseases such as hypertension, myocardial ischemia, myocardial hypertrophy, congestive heart failure, renal failure, asthma and cerebral vasospasm caused by subarachnoid hemorrhage, blocking the production or antagonism of endothelin The combination with the receptor can be used to treat the above diseases. The permeability of vascular endothelial cells and the transmembrane movement of cells are regulated by endothelin. Therefore, endothelin will affect the crossing of tumor cells to vascular endothelium, angiogenesis and blood-brain barrier. The study believes that endothelin receptor antagonists will become new drugs for the clinical treatment of hypertension, heart failure, subarachnoid hemorrhage, tumors, diabetes, kidney disease and renal failure, asthma and other diseases, especially for the treatment of Chronic congestive heart failure, reversal of cardiac hypertrophy, control of high blood pressure and prevention of stroke.

Contents of the invention

The purpose of the present invention is to provide a new class of pyrazole carboxylate endothelin receptor antagonists.

Another object of the present invention is to provide the application of the compound of general formula (I) in the preparation of medicines for treating cardiovascular and cerebrovascular diseases, tumors, diabetes, nephropathy, asthma, hyperthyroidism and other diseases.

The object of the present invention can be achieved through the following measures:

On the basis of the latest research on endothelin receptor antagonists, using the basic principles of drug design simulation, bioelectronic isosteres, and structural assembly, the compounds with general formula (I) were designed and synthesized, and their biological activities were carried out. Research. Among them, compounds I13 (code 0213) and I14 (code 0214) are particularly interesting.

Among them, R ₁ is C ₁ -C ₆ alkyl and alkenyl, carboxylic acid and its esters with one to four carbon atoms, aryl and substituted aryl with one to four carbon atoms, one to four A heterocyclic ring interspersed with carbon atoms; R ₂ and R ₃ are each independently H or halogen.

Wherein R ₁ can be C ₁ -C ₄ alkyl, R ₂ and R ₃ can be independently 4-chloro.

Wherein R ₁ can be n-butyl, R ₂ and R ₃ can be independently 4-chloro (code 0213).

Wherein R ₁ can be carboxylic acid and its esters with one to two carbon atom intervals, and R ₂ and R ₃ can be 4-chloro.

Wherein R ₁ can be carboxymethyl, R ₂ and R ₃ can be independently 4-chloro (code 0214).

The compound of general formula (I) included in the present invention can be made into various pharmaceutical dosage forms and pharmaceutical carriers.

The compound of the general formula (I) of the present invention is used as an endothelin receptor antagonist in the preparation of medicines for treating cardiovascular and cerebrovascular diseases, pulmonary hypertension, heart failure, tumors, diabetes, nephropathy and asthma.

The compound of general formula (I) can be synthesized according to table 1 or table 2 method:

Table 1 Reagents and conditions: i, ArCH ₂ Cl, K ₂ CO ₃ /acetone; ii, Ar'CH ₂ Cl, K ₂ CO ₃ /DMF; iii, (CO ₂ Et) ₂ , NaOEt/EtOH, reflux; iv , 85% NH ₂ NH ₂ ·H ₂ O/HOAc, reflux; v, R ₁ X, K ₂ CO ₃ /acetone, co-current; vi, NaOH/EtOH/H ₂ O, 10% HCl.

a: R ₁ =CH ₃ b: R ₁ =4-ClC ₆ H ₄ CH ₂

Table 2 Reagents and conditions: i, PhNHNH ₂ , HOAc, reflux; ii, 85% NH ₂ NH ₂ ·H ₂ O/HOAc, reflux; iii, CH ₃ I or 4-ClC ₆ H ₄ CH ₂ Cl, K ₂ CO ₃ /acetone, reflux;

2,4 disubstituted benzyloxyacetophenone (3) is condensed with diethyl oxalate to generate compound (4), and then cyclized with hydrazine hydrate to generate compound (5). However, the direct condensation of (7) with phenylhydrazine will generate two isomers (8) and (9). Similar isomers appear in the alkylation reaction of compound (5). For example, for the alkylation of compound (10), CH ₃ I with high activity and little steric hindrance is used as the alkylating agent to generate Almost the same amount of isomers (11a) and (12a), and the use of the activity ratio is small, and the p-chlorobenzyl chloride with the larger ratio of steric hindrance in the reaction process is used as the alkylation reagent, and the selectivity of the reaction is obviously improved. More than 90% of the product was isomer (11b). It can be seen that in the N-hydrocarbylation reaction of compound (5), the steric hindrance effect plays an important role, which causes the main product of this substitution reaction to be the A-type structure. The structural characteristics and physical and chemical constants of some of these isomer A or B-type compounds obtained by separation and purification are as follows:

and

compound R ₁ R ₂ R ₃ A or B Yield (%) Mp.(℃) 11a _CH3 h 4-Cl A 46 139-140 12a _CH3 h 4-Cl B 41 98-99 11b 4-ClC ₆ H ₄ CH ₂ h 4-Cl A 89 124-126 12b 4-ClC ₆ H ₄ CH ₂ h 4-Cl B 9.6 117-118 8 Ph 4-Cl 4-Cl A 3.4 121-122 9 Ph 4-Cl 4-Cl B 39 137-138

The physicochemical constants of general formula (I) representative compound are as follows:

^a Yield refers to the product yield of separation and purification

The final product can be separated and purified by column chromatography or refined by organic solvent recrystallization.

The preparation of pharmaceutical dosage forms using the compounds encompassed by the present invention as active ingredients requires only the use of conventional pharmaceutical formulation techniques. Tablets and capsules are preferred.

The compound of the present invention has strong endothelin receptor antagonistic activity and is used for treating cardiovascular disease and tumor patients. The dosage range is about 0.1-10 mg/kg/day.

The results of the endothelin receptor binding experiment of representative compounds of the present invention are as follows:

IC ₅₀ of Tab 1 compound to ET-1 binding assay compound IC ₅₀ (nM) I 01I 02I 03I 04I 05I 06I 07I 08I 09I 10I 11I 12I 13I 14I 15I 16I 17I 18I 19I 20I 21I 22I 23I 24I 25I 26I 27 280030001200500100010008001561>1000>10008001816>1000>1000>1000>1000>1000>10008995>1000500>1000>1000>1000

The activity of 0213 is similar to that of Bosentan approved by the US FDA in November 2001. The IC ₅₀ of Bosentan against ET _A R is 4.7nM, and that of ET _BR is 95nM. The activity of 0213 to antagonize ET _A R is about twice that of Bosentan, and the activity of antagonizing ET _BR is the same as that of the two drugs. The selectivity of Bosentan to ET _A R/ET _B R was 20.2. The selectivity of 0213 compound to ET _A R/ET _B R was 36.2, stronger than that of Bosentan.

The activity of compound 0213 against ET _A R and ET _B R is similar to that of compounds being developed by many famous foreign pharmaceutical companies.

Table 0213 Antagonism of ET _A R and ET _B R activity compared with IC ₅₀ of new compounds from foreign pharmaceutical companies compound ET _A R ET _B R selectivity 0213BosentanSB217242A182086BQ123TBC11251 2.574.71.194371.4 93951001.3-- 36.220.290.972.3 single-minded

1. The activity intensity of endothelin receptor antagonist 0213 on vascular _ETA and bronchial _ETB receptors

1.1 Anti-endothelin-1 (abbreviated as ET-1 or ET) on blood vessel activity, ET has a strong contractile activity on endothelialized thoracic aorta. All 77 compounds were quantitatively studied to antagonize ET contractile activity, and 9 compounds including 0213 were obtained as endothelin receptor antagonists with strong activity, among which 0213 had the strongest effect.

1.2 The ET receptor for contraction of endothelialized rat thoracic aorta is _ETA , and the contraction activity of endothelialized rat aortic ring induced by S6 _C is mediated by _ETB receptor. Results: The PA ₂ of 0213 compound against _ETA receptor was 8.52±0.26 (X±SD, the same below). The PA ₂ of antagonizing S6 _C- induced bronchoconstriction mediated by _ETB receptor was 6.56±0.20. The intensity of antagonizing _ETA is 91.2 times that of _ETB .

2. The activity analysis of endothelin receptor antagonist 0213 radioreceptor _{, the intensity of antagonizing ET and the activity intensity of ETA and ET B respectively} .

^125I -ET-1 competition for receptor ligand binding assay. When performing selective action on _ETAR , the _ETBR agonist _S6C was added to saturate the binding to _ETBR . For _ETBR selectivity, add _ETA selective blocker PD156707 to saturate and bind with _ETAR . It was obtained respectively: the IC ₅₀ of 0213 to ET _A R was 2.57±1.41nM (n=9), and the IC ₅₀ to ET _BR was 93±34nM (n=5), the activity difference between the two was 36 times. Tip: 0213 is a dual antagonist of ET _A R and ET _BR , mainly antagonizing ET _A R.

3. Experimental treatment of endothelin receptor antagonist 0213 on acute heart failure induced by coronary artery ligation in rats

0213 compound (100mg/kg, po.), 1 hour after administration to rats once, urethane 1.2g/kg was anesthetized, and the left coronary artery was ligated by thoracotomy. Measurement of hemodynamics: left ventricular systolic function: LV+dp/dtmax, LVSP; and left ventricular diastolic function: LV-dp/dtmin, LVDP. Observe for 120min.

After administration of 0213, acute heart failure significantly improved the reduction of LVSP, LV-dp/dtmin and LV+dp/dtmax. The mechanism of ET in acute heart failure is less, but there are still obvious drug effects, and it also has a slowing down of the heart rate (as shown in Figures 1, 2, and 3).

4. Chronic administration of endothelin receptor antagonist 0213, experimental treatment of rats with chronic congestive heart failure after myocardial infarction

After ligation of the left coronary artery in rats, a model of chronic congestive heart failure was established for 10d-70d. 0213 Establish 30mg/kg/d and 100mg/kg/d po for 60 days. The drug administration was started on the 11th day after coronary artery ligation, and the drug treatment was performed after heart failure was formed. Captopril 20mg/kg/d, 60 days. 10 days and 70 days after coronary artery ligation in rats, blood was taken to measure ANP (atrial natriuretic peptide), and the left ventricle and right ventricle were weighed and compared with body weight and hemodynamics. Comparing ANP, RV/BW, and LV/BW in the two pathological model groups, the changes in the model group at 10 and 70 days were significantly different from those in the sham operation group. Between 10 days and 70 days, LV+dp/dtmax, LVSP, LVDP, diastolic function, and systolic function also had significant differences, but hemodynamic LV-dp/dtmin and LVDP, and mean arterial blood pressure (MPA) and There was no significant difference in HR. In the tissue section of the pathological model, in the 10-day model of infarction, there was a large amount of inflammatory cell infiltration in the myocardial interstitium, and the inflammatory cell infiltration was significantly reduced in the 70-day model. Because ANP increased significantly and myocardial remodeling was more obvious in the 70d group, it suggested that in the 70d heart failure model, the humoral mechanism had been fully activated, and the myocardial remodeling and maintenance of heart failure were related to the humoral factors in the pathological mechanism.

4.1 Drug 0213 and captopril treated chronic congestive heart failure in rats, and the myocardial remodeling of congestive heart failure subsided.

Expressed as the ratio of myocardial weight to body weight of the declining heart: left ventricle/body weight (LV/BW), right ventricle/body weight (RV/BW), whole heart/body weight (LW/BW). Drug 0213 at 30mg/kg and 100mg/kg, po can significantly reduce left ventricular myocardial hypertrophy, and the drug effect is similar to that of captopril 20mg/kg, po. Compared with the sham operation group: the myocardial weight of the chronic heart failure 10-day group (CHF 10 days) increased significantly, indicating that the myocardial remodeling was obvious, ^b p<0.05, ^c p<0.01. Myocardial remodeling was more pronounced in the CHF70-day group. ^c p<0.01 compared with SO group. Compared with CHF 10 days, myocardial remodeling was more serious, ^f p<0.01. 0213 group significantly reduced myocardial remodeling, compared with CHF 70 days, both dose groups made regression, ^h p<0.05, or ⁱ p<0.01. The participating drug is captopril, which is also effective. (Figure 4)

4..2 Hemodynamic effects on congestive heart failure.

The systolic function LV+dp/dtmax of the heart with chronic failure decreased significantly, the diastolic function also failed obviously, and the increase of LV-dp/dtmin and LVDP was very significant. The hemodynamic LV+dp/dtmax, LV-dp/dtmin and LVDP of the failing heart were significantly improved in the drug 0213 and captopril groups (see Figures 5, 6, and 7).

4..3 Drug 0213 improves the increase of plasma ET.ANP concentration in congestive heart failure. Drug 0213 doses of 30 and 100mg/kg, po treatment, compared with rats with heart failure 70 days after infarction, significantly reduced the concentration of ET and ANP in plasma. (See Figure 8).

4.4 Changes of drug treatment on the mRNA expression of prepro-ET-1 and ANP in the pathological myocardium of rats with chronic heart failure. The expression of prepro-ET-1 mRNA in left ventricular myocardium of rats with chronic heart failure was significantly decreased, and the mRNA of ANP was significantly increased. (See Figure 9, 10)

The expression of mRNA in the myocardium of rats with chronic heart failure was significantly increased. In the 10-day and 70-day groups of myocardial infarction, significantly more than the sham operation group (SO). Compared with the sham operation group (SO): ^a p>0.05, ^b p<0.05, ^c p<0.01. Drug 0213 100mg/Kg group significantly decreased mRNA expression. Compared with 10 days of infarction: ^d p>0.05, ^e p<0.05, ^f p<0.01; compared with 70 days of infarction: ^g p>0.05, ^h p<0.05, ⁱ p<0.01.

The mRNA expression of Prepro-ET-1 was significantly increased in the 10-day infarction group compared with the sham operation group, ^c p<0.01, and also increased in the 70-day group, ^b p<0.05. The po of 0213100mg/Kg group made the expression of Prepro-ET-1 mRNA expression was significantly down-regulated. Compared with the sham operation group (SO): ^a p>0.05, ^b p<0.05, ^c p<0.01. Compared with 10 days of infarction: ^d p>0.05, ^e p<0.05, ^f p<0.01; compared with 70 days of infarction: ^g p>0.05, ^h p<0.05, ⁱ p<0.01.

The summary suggests that endothelin antagonist 0213 can be used in the treatment of acute and chronic heart failure with excellent curative effect.

5. Drug 0213 has obvious therapeutic effect on pulmonary hypertension. Significantly improved right ventricular systolic pressure (equivalent to pulmonary artery blood pressure) and subsided right ventricular myocardial remodeling in rats with pulmonary hypertension, and decreased plasma ET-1 and prepro-ET-1 and other indicators.

6. Drug 0213 improves thyroxine-induced cardiomyopathy and cardiovascular disease.

The drug 0213 can significantly improve the abnormal cardiovascular function of cardiomyopathy caused by multiple doses of L-thyroxine.

①Drug 0213 subsided 30.6% of myocardial remodeling induced by L-thyroxine (p<0.01).

②Reduce the concentration of ET-1 in the plasma of the cardiomyopathy model by 43.6% (p<0.01).

③ The mRNA of genes Kv1.4, Kv4.2, and Kv4.3 in the diseased myocardium was significantly down-regulated, and the drug 0213 treatment significantly increased it to normal.

④ The drug 0213 significantly improved the myocardial lesion of cardiomyopathy under the electron microscope.

⑤ iNOS activity increased in thyroxin-induced cardiomyopathy, and the drug 0213 decreased it by 82.5% (p<0.01). The eNOS activity decreased in this cardiomyopathy, and the drug 0213 increased the eNOS activity by 87.2%, (p<0.01).

⑥ In thyroxin-induced cardiomyopathy, cNOS was significantly down-regulated, and the drug 0213 made it up-regulated by 100%, completely returning to normal (p<0.01). The mRNA of iNOS was significantly up-regulated in cardiomyopathy, and the drug 0213 increased the expression of iNOS mRNA by 100%, p<0.01, completely returning to normal.

⑦ NO in the myocardium of thyroxin-induced cardiomyopathy decreased significantly, and the drug 0213 increased it by 68%, (p<0.01).

⑧ The apoptosis rate of cardiomyocytes in thyroxin-induced cardiomyopathy was 97.5% (p<0.01). The aneuploidy rate of cardiomyocytes in cardiomyopathy reaches 13.5%, and the aneuploidy completely disappears after treatment with drug 0213.

⑨Drug 0213 treatment can significantly improve the vascular disease caused by thyroxine.

Summary Endothelin antagonist 0213 has a significant therapeutic effect on cardiomyopathy and vascular lesions caused by large doses of thyroxine, suggesting that endothelin antagonist 0213 can be used to treat cardiomyopathy and hyperthyroidism.

7 Drug 0214 for the treatment of high blood pressure

For the hypertension caused by DOCA plus drinking saline, the acute iv administration and chronic gavage of the drug 0214 can significantly reduce the blood pressure in the DOCA hypertension model.

In foreign literature, many experimental and clinical studies have been reported on the treatment of left heart failure (heart failure), pulmonary hypertension, bronchial asthma, and subarachnoid hemorrhage with endothelin receptor antagonists. Currently, the FDA is the first to approve Bosentan for the treatment of pulmonary arterial hypertension. The hypertrophy of the pulmonary artery wall and the weakening of the right heart caused by pulmonary arterial hypertension are all involved in the mechanism of endothelin. 0213 has curative effect on left heart failure and pulmonary hypertension. 0213 is effective for diseases caused by increased endothelin, such as cardiomyopathy and hyperthyroidism. The above is an example of the biological activity of the neoendothelin antagonist 0213 compound.

Description of drawings

Fig. 1 is the experimental treatment diagram of drug 0213 of the present invention obviously improving LVSP of acute heart failure (AHF) in rats.

Fig. 2 is an experimental treatment diagram of the medicine 0213 of the present invention obviously improving LV+dp/dt _max of acute heart failure (AHF) in rats.

Fig. 3 is an experimental treatment diagram of the medicine 0213 of the present invention obviously improving LV-dp/dt _min of acute heart failure (AHF) in rats.

Fig. 4 is an experimental treatment diagram illustrating that the drug 0213 of the present invention makes myocardial remodeling subside in rats with chronic heart failure.

Fig. 5 is the experimental treatment diagram of the medicine 0213 of the present invention obviously improving the LV+dp/dt _max of the failing heart.

Fig. 6 is an experimental treatment diagram of drug 0213 of the present invention, which significantly improves LV-dp/dt _min of failing heart.

Fig. 7 is the experimental treatment diagram of drug 0213 of the present invention, which obviously improves LVDP of failing heart.

Fig. 8 is an experimental treatment diagram of drug 0213 of the present invention in reducing ET and ANP in rats with heart failure.

Fig. 9 is an experimental treatment chart of improving ANP mRNA expression in chronic heart failure rats treated with drug 0213 of the present invention.

Fig. 10 is an experimental treatment chart of improving preproET-1 mRNA expression in rats with chronic heart failure treated with drug 0213 of the present invention.

Detailed ways

The following examples can make those skilled in the art understand the present invention more comprehensively, but the present invention cannot be limited in any way.

Example

4-Benzyloxy-2-hydroxyacetophenone

Dissolve 5g (0.033mol) 2,4-dihydroxyacetophenone in 150ml acetone, add 6.5g (0.05mol) K ₂ CO ₃ , 3g KI, 0.1g TBAB, and slowly drop in 4.9g ( 0.039mol) benzyl chloride, reacted at room temperature for 0.5hr, then refluxed for 2hr, cooled to room temperature, filtered, the filtrate was evaporated to dryness, and the residue was recrystallized with absolute ethanol to obtain 5.6g of white needle crystals, with a yield of 70%.

4-(p-chlorobenzyloxy)-2-hydroxyacetophenone

Dissolve 7.6g (0.05mol) of 2,4-dihydroxyacetophenone in 150ml of acetone, add 7g (0.05mol) of K ₂ CO ₃ , 3g of KI, 0.1g of TBAB and slowly drop into 7.7g (0.05 mol) p-chlorobenzyl chloride was reacted at room temperature for 0.5 hr, then refluxed for 3 hr, cooled to room temperature, filtered, the filtrate was evaporated to dryness, and the residue was recrystallized with absolute ethanol to obtain 10.4 g of white needle crystals with a yield of 75%.

2,4-bis(p-chlorobenzyloxy)acetophenone

Mix 15.2g (0.1mol) 2,4-dihydroxyacetophenone, 40g (0.25mol) p-chlorobenzyl chloride, 55g (0.4mol) K ₂ CO ₃ in 70ml DMF, and react at 50°C for 6hr , cooled to room temperature, the reaction solution was poured into ice water, and the precipitated solid was recrystallized with absolute ethanol to obtain 24 g of white crystals, with a yield of 60%.

1-Benzyloxy-4-(p-chlorobenzyloxy)acetophenone

React 5g (0.018mol) 4-(p-chlorobenzyloxy)-2-hydroxyacetophenone with 3g (0.025mol) benzyl chloride, 5g ( _0.036mol ) _K2CO3 in 50ml DMF at 50°C After 5 hours, cool to room temperature, pour into ice water, and recrystallize the precipitated solid with absolute ethanol to obtain 3.6 g of light brown needle crystals, with a yield of 56%.

2,4-Di(p-chlorobenzyloxy)phenacylmethyl oxalate diethyl ester

23g (0.057mol) of 2,4-bis(p-chlorobenzyloxy)acetophenone was dissolved in 80mlTHF, and 18g (0.078mol) of diethyl oxalate was added dropwise at 50°C. ) the 100ml sodium ethylate solution that metal sodium is made, dripping finishes, be warming up to reflux reaction 8hr, cooling, pour into the 500ml water that contains concentrated HCl, the solid that separates out is filtered, washes with ethanol, and drying obtains 28g khaki solid, yield 98 %.

3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid ethyl ester

Dissolve 25g (0.05mol) of 2,4-bis(p-chlorobenzyloxy)phenacylmethyl oxalate diethyl oxalate prepared above in 200ml of glacial acetic acid, and slowly add 3.6g (0.06mol) of 85% Hydrazine hydrate, dropwise, reflux reaction for 8hr, cooled to room temperature, white needle crystals precipitated, filtered, washed with HOAc recrystallized absolute ethanol, dried to obtain 19.2g of white powdery solid, yield 77%, mp: 177-179 ℃. ¹ HNMR (CDCl ₃ ) δ1.32 (3H, t, -CO ₂ CH ₂ CH ₃ ), 4.28 (2H, q. -CO ₂ CH ₂ CH ₃ ), 5.12 (2H, s, 4-OCH ₂ Ar) , 5.24 (2H, s, 2-OCH ₂ Ar), 6.68 (1H, d, J=9.0, 5-H), 6.79 (1H, s, 3-H), 7.03 (1H, s, 4'-H of pyrazole), 7.40-7.59 (8H, m, -ArH), 7.71 (1H, d, J=8.6, 6-H), 8.20 (1H, s, -NH).

3-(2-Benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid ethyl ester

Referring to the preparation method of ethyl 3-(2,4-bis(p-chlorobenzyloxy)phenyl)-pyrazole-5-carboxylate, a white solid was obtained with a yield of 72%, mp: 137-138°C.

3-(2-Benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I01)

Dissolve 2g (4.3mmol) of the above compound in 15ml of THF, add 25ml of methanol and 10ml of 10% KOH, reflux for 2 hours, cool to room temperature, adjust pH=3 with 10% HCl, add appropriate amount of water to dilute, and filter the obtained solid with Recrystallization from DMF-EtOH gave 1.0 g of a colorless granular solid with a yield of 53%. mp: 239-241°C. Anal. C ₂₄ H ₁₉ ClN ₂ O ₄ ·0.5H ₂ O. Calcd: C, 64.94; H, 4.51; N, 6.31. Found: C, 65.26; H, 4.31; N.6.43 .IR(KBr)ν3249, 2866, 1712, 1614, 1580, 1493, 1455, 1410, 1276, 1301, 1279, 1239, 1170, 1058, 1005, 962, 807, 754, 700cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ )δ5.06 (2H, s, 4-OCH ₂ Ar), 5.20 (2H, s, 2-OCH ₂ Ph), 6.66 (1H, d, J=8.6, 5-H), 6.70 (1H, d, J=2.0, 3-H), 7.10 (1H, s, 4'-H of pyrazole), 7.34-7.45 (9H, m, -PhH and ArH), 7.66 (1H, d, J= 8.6, 6-H). MS (EI) m/e: 434 (M ⁺ +1), 343, 125, 91 (100).

1-Ethyl-3-(2-benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid ethyl ester

Dissolve 1g (2mmol) ethyl 3-(2-benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylate in 20ml acetone, add 2g (14mmol) K ₂ CO ₃ and 1ml of bromoethane, reflux for 2 hours, cool to room temperature, filter, distill off an appropriate amount of acetone, add methanol, and precipitate 0.76 g of colorless needle crystals, yield 75%, mp: 130-131°C.

1-Ethyl-3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid ethyl ester

Referring to the preparation method of compound 1-ethyl-3-(2-benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid ethyl ester, white needle crystals were obtained with a yield of 81 %, mp: 95-96°C. ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ1.34 (3H, t, -CO ₂ CH ₂ CH ₃ ), 1.46 (3H, t, -NCH ₂ CH ₃ ), 4.30 (2H, q, -CO ₂ CH ₂ CH ₃ ), 4.62 (2H, q, -NCH ₂ CH ₃ ), 5.01 (2H, s, 4-OCH ₂ Ar), 5.06 (2H, s, 2-OCH ₂ Ar), 6.58 (1H, d , J=2.1, 3-H), 6.62 (1H, dd, J=2.1 and 9.0, 5-H), 7.21 (1H, s.4'-H of pyrazole), 7.28-7.51 (8H, m, - ArH), 7.92 (1H, d, J=9.0, 6-H).

1-methyl-3-(2-benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I02)

Referring to the preparation of compound I03, white crystals were obtained with a yield of 61%. mp: 203-204°C. Anal. C ₂₅ H ₂₁ ClN ₂ O _{4 .} Calcd: C, 66.89; H, 4.68; N, 6.24. Found: C, 66.67; H, 4.68; N, 6.48. IR (KBr) ν 3029, 2913, 1696, 1612, 1586, 1539, 1489, 1451, 1283, 1249, 1184, 1108, 1013, 810, 767, 700cm ^-1 ; ¹ HNMR(CDCl ₃ +DMSO-d ₆ ) δ4.20( 3H, s, -NCH ₃ ), 5.03 (2H, s, -OCH ₂ -), 5.17 (2H, s, -OCH ₂ -), 6.61 (1H, dd, J=2.4 and 8.7, 5-H), 6.63 (1H, d, J = 2.3, 3-H), 7.30 (1H, s, 4'-H of pyrazole), 7.31-7.46 (9H, m, -ArH and PhH), 7.86 (1H, d, J =8.8, 6-H). MS (EI) m/e: 448 (M ⁺ +1), 403, 357, 125.91 (100).

1-Ethyl-3-(2-benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I03)

Dissolve 0.55g (1mmol) of the above-mentioned compound 1-ethyl-3-(2-benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid ethyl ester in 15ml THF and 10ml To MeOH, 10ml of 10% NaOH was added, refluxed for 0.5hr, cooled to room temperature, adjusted to pH=3 with 10% HCl, and the obtained solid was recrystallized with absolute ethanol to obtain 0.35g of white needle crystals, with a yield of 64%. mp: 200-201°C. Anal. C ₂₆ H ₂₃ ClN ₂ O _{4 .} Calcd: C, 67.46; H, 4.97; N, 6.05. Found: C, 67.35; H, 5.05; ν2979, 1694, 1612, 1587, 1536, 1451, 1301, 1281, 1237, 1183, 1110, 1060, 1013, 810, 768, 700cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ1.52 (3H , t, -NCH ₂ CH ₃ ), 4.69 (2H, q, -NCH ₂ CH ₃ ), 5.05 (2H, s, 4-OCH ₂ Ar), 5.18 (2H, s, 2-OCH ₂ Ph), 6.65 -6.67 (2H, m, 3-H and 5-H), 7.26-7.47 (10H, m, -ArH, PhH and 4'-H of pyrazole), 7.96 (1H, d, J=9.1, 6-H ). MS (EI) m/e: 462 (M ⁺ +1), 417, 337, 125, 91 (100).

1-Propyl-3-(2-benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I04)

Referring to the preparation of compound I03, white crystals were obtained with a yield of 50%. mp: 182-184°C. Anal. C ₂₇ H ₂₅ ClN ₂ O _{4 .} Calcd: C, 68.00; H, 5.25; N, 5.88. Found: C, 68.27; H, 5.05; N, 5.81. IR (KBr) ν3438, 3032, 2966, 1693, 1614, 1585, 1535, 1453, 1376, 1290, 1175, 1126, 1062, 1015, 810, 739 cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ0.96 (3H , t, -NCH ₂ CH ₂ CH ₃ ), 1.91 (2H, 6, -NCH ₂ CH ₂ CH ₃ ), 4.56 (2H, t, -NCH ₂ CH ₂ CH ₃ ), 5.03 (2H, s, 4- OCH ₂ Ar), 5.16 (2H, s, 2-OCH ₂ Ph), 6.62 (1H, dd, J=2.2 and 9.2, 5-H), 6.64 (1H, d, J=2.1, 3-H), 7.31-7.46 (10H, m, -ArH, PhH and 4'-H of pyrazole), 7.91 (1H, d, J=9.1, 6-H). MS (EI) m/e: 476 (M ⁺ +1 ), 431, 351, 125, 91(100).

1-Butyl-3-(2-benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I05)

Referring to the preparation of compound I03, white crystals were obtained with a yield of 88%. mp: 170-172°C. Anal. C ₂₈ H ₂₇ ClN ₂ O ₄ . Calcd: C, 68.50; H, 5.50; N, 5.71. Found: C, 68.68; H, 5.56; ν2961, 1693, 1614, 1540, 1448, 1288, 1175, 1125, 1062, 1015, 812 cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ0.93 (3H, t, -CH ₂ CH ₃ ), 1.37 (2H, 6, -N(CH ₂ ) ₂ CH ₂ CH ₃ ), 1.85 (2H, 5, -NCH ₂ CH ₂ CH ₂ CH ₃ ), 4.60 (2H, t, -NCH ₂ (CH ₂ ) ₂ CH ₃ ), 5.01 (2H, s, 4-OCH ₂ Ar), 5.16 (2H, s, 2-OCH ₂ Ph), 6.59-6.63 (2H, m, 3-H and 5-H), 7.29 (1H , s, 4'-H of pyrazole), 7.30-7.44 (9H, m, -ArH and PhH), 7.89 (1H, d, J=9.2, 6-H). MS (EI) m/e: 490 ( M ⁺⁺ 1), 445, 365, 125, 91 (100).

1-(3-carboxypropyl)-3-(2-benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I06)

Referring to the preparation of compound I03, white crystals were obtained with a yield of 75%. mp: 202-203°C. Anal. C ₂₈ H ₂₅ ClN ₂ O ₆ . Calcd: C, 64.55; H, 4.80; N, 5.38. Found: C, 64.75; H, 4.82; N, 5.36.IR(KBr) ν3062, 2937, 1710, 1692, 1583, 1540, 1450, 1292, 1240, 1171, 1102, 1016, 810, 768cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ2.21 (2H, p, - NCH ₂ CH ₂ CH ₂ CO ₂ H), 2.35 (2H, t, -N(CH ₂ ) ₂ CH ₂ CO ₂ H), 4.67 (2H, t, -NCH ₂ -), 5.02 (2H, s, - OCH ₂ Ar), 5.16 (2H, s, -OCH ₂ Ph), 6.59-6.63 (2H, m, 3-H and 5-H), 7.30-7.43 (10H, m, -ArH, PhH and H of pyrazole ), 7.89 (1H, dJ=9.2, 6-H). MS (EI) m/e: 520 (M ⁺ +1), 476, 351.125 (100), 91.

1-Benzyl-3-(2-benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I07)

Referring to the preparation of compound I03, white crystals were obtained with a yield of 75%. mp: 217-218°C. Anal. C ₃₁ H ₂₅ ClN ₂ O _{4 .} Calcd: C, 70.92; H, 4.77; N, 5.34. Found: C, 70.86; H, 4.67; N, 5.28. IR (KBr) ν3031, 2952, 1691, 1610, 1582, 1535, 1452, 1381, 1295, 1272, 1183, 1095, 1012, 822, 768, 717cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ5.05 (2H , s, -OCH ₂ Ar), 5.18 (2H, s, -OCH ₂ Ph), 5.81 (2H, s, -NCH ₂ Ph), 6.64-6.66 (2H, m, 3-H and 4-H), 7.26-7.46 (10H, m, -ArH, PhH and4'-H of pyrazole), 7.99 (1H, d, J=9.3, 6-H). MS (EI) m/e: 524 (M ⁺ +1) , 433, 125, 91(100).

1-(4-chlorobenzyl)-3-(2-benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I08)

Referring to the preparation of compound I03, white crystals were obtained with a yield of 51%. mp: 211-212°C. Anal. C ₃₁ H ₂₄ Cl ₂ N ₂ O ₄ .Calcd: C, 66.67; H, 4.30; N, 5.02.Found: C, 66.47; H, 4.36; N, 5.19.IR( KBr) ν3027.1693, 1613.1583, 1492, 1450, 1292, 1173, 1092, 1072, 1016, 810, 696cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ5.02 (2H, s, -OCH ₂ Ar), 5.16 (2H, s, -OCH ₂ Ph), 5.77 (2H, s, -NCH ₂ Ar), 6.59-6.63 (2H, m, 3-H and 4-H), 7.23-7.43 (14H, m, -ArH, PhH and4'-H of pyrazole), 7.92 (1H, d, J=9.0, 6-H). MS (EI) m/e: 558 (M ⁺⁺ 1), 433, 125, 91 (100).

3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I09)

Referring to the preparation method of compound I01, a colorless crystal was obtained with a yield of 82%. mp: 236-238° C. Anal. C ₂₄ H ₁₈ Cl ₂ N ₂ O ₄ . Calcd: C, 61.41; H, 3.84; N, 5.97. Found: C, 61.40; H, 3.90; N, 6.29.IR( KBr) ν3235, 2896, 1713, 1614, 1580, 1492, 1453, 1408, 1376, 1279, 1239, 1171, 1061, 1005, 963, 801cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ2.65 (1H, br, -NH), 5.06 (2H, s, -OCH ₂ Ar), 5.15 (2H, s, -OCH ₂ Ar), 6.65 (1H, d, J=2.3, 3-H), 6.67 ( 1H, d, J=2.4 and 9.2, 5-H), 7.09 (1H, s, 4'-H of pyrazole), 7.32-7.41 (8H, m, -ArH), 7.65 (1H, d, J=9.3 , 6-H). MS (EI) m/e: 468 (M ⁺ +1), 423, 125 (100).

1-Methyl-3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I10)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 71%. mp: 205-206°C. Anal. C ₂₅ H ₂₀ Cl ₂ N ₂ O ₄ .Calcd: C, 61.98; H, 4.13; N, 5.79.Found: C, 62.21; H, 4.18; N, 6.05.IR( KBr) ν1694.1615, 1584, 1539, 1492, 1450, 1368, 1293, 1182, 1116, 1014, 808, 767cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ4.21 (3H, s, - NCH ₃ ), 5.02 (2H, s, -OCH ₂ Ar), 5.12 (2H, s, -OCH ₂ Ar), 6.57 (1H, d, J=2.4, 3-H), 6.63 (1H, d, J =2.4and 8.6, 5-H), 7.28 (1H, s, 4'-H of pyrazole), 7.30-7.38 (8H, m, -ArH), 7.86 (1H, d, J=8.6, 6-H) .MS(EI) m/e: 482(M ^- +1), 437.357, 313, 125(100).

1-Ethyl-3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I11)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 90%. mp: 192-193° C. Anal. C ₂₆ H ₂₂ Cl ₂ N ₂ O ₄ . Calcd: C, 62.78; H, 4.43; N, 5.63. Found: C, 63.24; H, 4.58; N, 5.95.IR( KBr) ν2982, 2877, 1693, 1578.1537, 1492, 1455, 1408, 1277, 1182, 1088, 1054, 1014, 815cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ1.43 (3H, t, - NCH ₂ CH ₃ ), 4.59 (2H, q, -NCH ₂ CH ₃ ), 5.04 (2H, s, -OCH ₂ Ar), 5.13 (2H, s, -OCH ₂ Ar), 6.57-6.66 (2H, m , 3-H and5-H), 7.20 (1H, s, 4'-H of pyrazole), 7.27-7.45 (8H, m, -ArH), 7.85 (1H, d, J=9.0, 6-H). MS(EI) m/e: 496(M ⁺ +1), 423, 371, 327, 125(100).

1-Propyl-3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I12)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 83%. mp: 184-186° C. Anal. C ₂₇ H ₂₄ Cl ₂ N ₂ O ₄ . Calcd: C, 63.41; H, 4.70; N, 5.48. Found: C, 63.52; H, 4.75; N, 5.73.IR( KBr) ν2966, 2935, 1692, 1614, 1584, 1539, 1491, 1452, 1376, 1286, 1178, 1123, 1087, 1061, 1014, 810, 767cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ0 .96 (3H, t, -N(CH ₂ ) ₂ CH ₃ ), 1.93 (2H, h, -NCH ₂ CH ₂ CH ₃ ), 4.58 (2H, t, -NCH ₂ CH ₂ CH ₃ ), 5.05 ( 2H, s, -OCH ₂ Ar), 5.14 (2H, s, -OCH ₂ Ar), 6.60 (1H, d, J=2.3, 3-H), 6.65 (1H, dd, J=2.3 and 8.6, 5 -H), 7.29 (1H, s, 4'-H of pyrazole), 7.34-7.41 (8H, m, -ArH), 7.89 (1H, d, J=8.5, 6-H).MS(EI)m /e: 510(M ⁺⁺ 1), 466, 385, 125(100).

1-Butyl-3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I13)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 78%. mp: 164-165° C. Anal. C ₂₈ H ₂₆ Cl ₂ N ₂ O ₄ . Calcd: C, 64.00; H, 4.95; N, 5.33. Found: C, 63.62; H, 5.13; N, 5.40.IR( KBr) ν2960, 2872, 1692, 1614, 1583, 1492, 1451, 1375, 1283, 1179, 1120, 1090, 1015, 810cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ0.93 (3H, t , -N(CH ₂ ) ₃ CH ₃ ), 1.34 (2H, h, -NCH ₂ CH ₂ CH ₂ CH ₃ ), 1.84 (2H, p, -NCH ₂ CHH ₂ CH ₂ CH ₃ ), 4.57 (2H, t, -NCH ₂ (CH ₂ ) ₂ CH ₃ ), 5.00 (2H, s, -OCH ₂ Ar), 5.09 (2H, s, -OCH ₂ Ar), 6.56-6.62 (2H, m, 3-H and 5-H), 7.23 (1H, s, 4'-H of pyrazole), 7.30-7.45 (8H, m, -ArH), 7.87 (1H, d, J=9.0, 6-H).MS(EI) m/e: 524(M ⁺⁺ 1), 480, 399, 355, 125(100).

1-carboxymethyl-3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I14)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 33%. mp: 210-212°C. Anal. C ₂₆ H ₂₀ Cl ₂ N ₂ O ₆ .Calcd: C, 59.20; H, 3.80; N, 5.31.Found: C, 59.17; H, 3.92; N, 5.72.IR( KBr)ν 2952, 1735, 1695, 1614, 1583 _, 1492, 1450 ^, 1408, 1292, 1257, 1176, 1092, _1015, 830, 806cm- ¹ ; 2H, s, -OCH ₂ Ar), 5.12 (2H, s, -OCH ₂ Ar), 5.34 (2H, s, -NCH ₂ CO ₂ H), 6.58 (1H, d, J=2.0, 3-H) , 6.61 (1H, dd, J=2.0 and8.5, 5-H), 7.32-7.39 (9H, m, -ArH and 4'-H of pyrazole), 7.87 (1H, d, J=8.4, 6- H). MS (EI) m/e: 526 (M ⁺ +1), 482, 401, 357, 125 (100).

1-(3-carboxypropyl)-3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I15)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 63%. mp: 202-204°C. Anal. C ₂₈ H ₂₄ Cl ₂ N ₂ O ₆ . Calcd: C, 60.54; H, 4.32; N, 5.05. Found: C, 60.53; H, 4.53; N, 5.31.IR( KBr) ν3036, 2930, 1691, 1613, 1583, 1537, 1492, 1452, 1288, 1232, 1171, 1097, 1016, 811cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ2.20 (2H, p , -NCH ₂ CH ₂ CH ₂ CO ₂ H), 2.34(2H, t, -N(CH ₂ ) ₂ CH ₂ CO ₂ H), 4.66(2H, t, -NCH ₂ (CH ₂ ) ₂ CO ₂ H ), 5.02 (2H, s, -OCH ₂ Ar), 5.11 (2H, s, -OCH ₂ Ar), 6.57 (1H, d, J=2.2, 3-H), 6.62 (1H, dd, J=2.2 and 8.7, 5-H), 7.27 (1H, s, 4'-H of pyrazole), 7.31-7.39 (8H, m, -ArH), 7.87 (1H, d, J=8.5, 6-H).MS (EI)m/e: 554(M ⁺ +1), 510, 430, 385, 125(100).

1-(2-Bromoethyl)-3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I16)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 71%. mp: 190-191° C. Anal. C ₂₆ H ₂₁ BrCl ₂ N ₂ O _{4 .} Calcd: C, 54.17; H, 3.65; N, 4.86. Found: C, 54.07; H, 3.43; N, 4.88.IR( KBr) ν3027, 2952, 2874, 1693, 1614, 1585, 1492, 1452, 1407, 1294, 1178, 1091, 1014, 807, 766cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ4.04 (2H , t, -CH ₂ Br), 4.77 (2H, t, -NCH ₂ -), 5.03 (2H, s, -OCH ₂ Ar), 5.12 (2H, s, -OCH ₂ Ar), 6.59 (1H, d , J=2.4, 3-H), 6.63 (1H, dd, J=2.4 and 8.4, 5-H), 7.30 (1H, s, 4'-H of pyrazole), 7.32-7.36 (8H, m, - ArH), 7.86 (1H, d, J=8.6, 6-H). MS (EI) m/e: 574 (M ⁺ +1), 494, 422, 369, 125 (100).

1-(2-Pyrrolidin-1-yl)ethyl-3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I17)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 88%. mp: 227-228°C. Anal. C ₃ 0H ₁₉ Cl ₂ N ₃ O ₄ .Calcd: C, 63.60; H, 5.12; N, 7.42.Found: C, 63.47; H, 5.33; N, 7.49.IR( KBr) ν3655, 3395, 1610, 1581, 1533, 1493, 1453, 1405, 1376, 1297, 1183, 1088, 1012, 814, 789cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ1.46-2.07 (4H, br, -CH ₂ (CH ₂ ) ₂ CH ₂ -of pyrrolidine), 2.20 (4H, br, -CH ₂ (CH ₂ ) ₂ CH ₂ - of pyrrolidine), 3.71 (2H, t, -NCH ₂ CH ₂ -pyrrrolidine), 5.07 (2H, s, -OCH ₂ Ar), 5.20 (2H, s, -OCH ₂ Ar), 5.26 (2H, t, -NCH ₂ CH ₂ -pyrrolidine), 6.60 (1H, d , J=2.1, 3-H), 6.67 (1H, dd, J=2.3 and 8.6, 5-H), 7.28 (1H, s, 4'-H of pyrazole), 7.34-7.44 (8H, m, - ArH), 7.86 (1H, d, J=8.6, 6-H). MS (EI) m/e: 565 (M ⁺ +1), 125, 97, 84 (100).

1-allyl-3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I18)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 75%. mp: 190-192° C. Anal. C ₂₇ H ₂₂ Cl ₂ N ₂ O ₄ . Calcd: C, 63.65; H, 4.32; N, 5.50. Found: C, 63.51; H, 4.26; N, 5.57.IR( KBr) ν2877, 1692, 1610, 1578, 1536, 1492, 1456, 1408, 1275, 1182, 1093, 1014, 938, 813 cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ5.02 (2H, s , -OCH ₂ Ar), 5.11 (2H, s, -OCH ₂ Ar), 5.09-5.18 (2H, m, -CH=CH ₂ ), 5.23 (2H, d, J=5.58, -NCH ₂ CH=CH ₂ ), 6.08 (1H, m, -CH=CH ₂ ), 6.58 (1H, d, J=2.3, 3-H), 6.62 (1H, dd, J=2.3 and 8.5, 5-H), 7.29 ( 1H, s, 4'-Hof pyrazole), 7.31-7.38 (8H, m, -ArH), 7.87 (1H, d, J=8.4, 6-H). MS (EI) m/e: 508 (M ⁺ +1), 464, 383, 125 (100).

1-phenyl-3-(2,4-di(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I19)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 66%. mp: 206-208°C. Anal. C ₃₀ H ₂₂ Cl ₂ N ₂ O ₄ .Calcd: _C , 66.06; _H , 4.04; _N , 5.14.Found: _C , 65.67; _H , 4.45; _N , 5.13.IR( KBr) ν 2919, 1697, 1617, 1555, 1492, 1451, 1372, 1311, 1230, 1181, 1127, 1015, 812, 776.694cm ^-1 ; ¹ HNMR (CDCl ₃ ) δ5.05 (2H, s, -OCH ₂ Ar), 5.16 (2H, s, -OCH ₂ Ar), 6.64-6.66 (2H, m, 3-H and 5-H), 7.28-7.55 (9H, m, ArH and 4'-H of pyrazole) , 7.98 (1H, d, J=8.1.6-H). MS (EI) m/e: 544 (M ⁺ +1), 500, 375, 125 (100).

1-Benzyl-3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I20)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 87%. mp: 187-188°C. Anal. C ₃₁ H ₂ 4Cl ₂ N ₂ O ₄ ·0.5H ₂ O. Calcd: C, 65.49; H, 4.40; N, 4.93. Found: C, 65.11; H, 4.24; N .5.09.IR(KBr)ν3030, 2948, 2867, 1695, 1612, 1582, 1541, 1493, 1450, 1282, 1178, 1095, 1015, 811, 722cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ5.03 (2H, s, -OCH ₂ Ar), 5.11 (2H, s, -OCH ₂ Ar), 5.82 (2H, s, -NCH ₂ Ph), 6.58 (1H, d, J=2.3, 3- H), 6.62 (1H, dd, J=2.3and 8.6, 5-H), 7.22-7.38 (14H, m, -PhH, ArH and 4'-H of pyrazole), 7.91 (1H, d, J=8.5 , 6-H). MS (EI) m/e: 558 (M ⁺ +1), 467, 433, 389, 125 (100), 91.

1-(4-chlorobenzyl)-3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I21)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 98%. mp: 233-234° C. Anal. C ₃₁ H ₂₃ Cl ₃ N ₂ O _{4 .} Calcd: C, 62.68; H, 3.88; N, 4.72. Found: C, 63.03; H, 3.94; N, 5.07.IR( KBr) ν 3031, 2954, 1692, 1614, 1586, 1492, 1454, 1408, 1279, 1184, 1093, 1015, 811, 748cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ5.03 (2H, s, -OCH ₂ Ar), 5.11 (2H, s, -OCH ₂ Ar), 5.77 (2H, s, -NCH ₂ Ar), 6.58 (1H, d, J=2.3, 3-H), 6.62 (1H , dd, J=2.3 and 8.5, 5-H), 7.24-7.41 (13H, m.-ArH and 4'-H of pyrazole), 7.90 (1H, d, J=8.5, 6-H).MS( EI) m/e: 592 (M ⁺ +1), 467, 125 (100), 91.

1-Benzyl-3-(2,4-bis(p-fluorobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I22)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 68%. mp: 203-204°C. Anal. C ₃₁ H ₂₄ F ₂ N ₂ O _{4 .} Calcd: C, 70.71; H, 4.59; N, 5.32. Found: C, 70.29; H, 4.12; N, 5.97.

1-Benzyl-3-(2,4-di(o-iodobenzyloxy)phenyl)pyrazole-5-carboxylic acid (I23)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 82%. mp: 197-198°C. Anal. C ₃₁ H ₂₄ I ₂ N ₂ O _{4 .} Calcd: C, 50.16; H, 3.26; N, 3.77. Found: C, 50.58; H, 3.77; N, 4.12.

1-Ethyl-3-(2-o-bromobenzyloxy, 4-p-iodobenzyloxy)phenyl-pyrazole-5-carboxylic acid (I24)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 84%. mp: 200-201 ° C. Anal. C ₂₆ H ₂₂ Cl ₂ N ₂ O ₄ . Calcd: C, 49.31; H, 3.50; N, 4.42. Found: C, 49.80; H, 3.78; N, 4.88.1- Ethyl-3-(2-o-chlorobenzyloxy, 4-p-bromobenzyloxy)phenyl-pyrazole-5-carboxylic acid (I25)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 79%. mp: 198-199°C. Anal _{. C26H22Cl2N2O4} . _{Calcd :} C, 57.63; H, 4.09; N, 5.17 _. Found: C, 58.01; H, 4.41; N, 5.45.

1-methyl-5-(2-benzyloxy-4-(4-chlorobenzyloxy)phenyl)pyrazole-3-carboxylic acid (I26)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 89%. mp: 148-150°C. Anal. C ₂₅ H ₂₁ ClN ₂ O _{4 .} Calcd: C, 66.89; H, 4.68; N, 6.24. Found: C, 66.53; H, 4.98; N, 6.46. IR (KBr) ν 3029, 2897, 1693, 1615, 1580, 1556, 1491, 1452, 1372, 1306, 1250, 1173, 1117, 1012, 807.731, 694cm ^-1 ; ¹ HNMR (CDCl ₃ +DMSO-d ₆ ) δ3.72( 3H, s, -NCH ₃ ), 5.11 (2H, s, -OCH ₂ Ar), 5.12 (2H, s, -OCH ₂ Ar), 6.65 (1H, s, 4'-H of pyrazole), 6.68 (1H , dd, J=2.1 and 8.5, 5-H), 6.80 (1H, d, J=2.1, 3-H), 7.19 (1H, d, J=8.4, 6-H), 7.26-7.33 (5H, m, -PhH), 7.37-7.46 (4H, m, -ArH). MS (EI) m/e: 448 (M ⁺ +1), 404, 125, 91 (100).

1-Phenyl-5-(2,4-bis(4-chlorobenzyloxy)phenyl)pyrazole-3-carboxylic acid (I27)

Referring to the preparation method of compound I03, white crystals were obtained with a yield of 67%. mp: 205-206°C. Anal. C ₃₀ H ₂₂ Cl ₂ N ₂ O ₄ ·0.5H ₂ O. Calcd: C, 67.15; H, 3.97; N, 5.05. Found: C, 67.13; H, 4.23; N , 5.53.IR (KBr) ν3067, 2922, 2872, 1705, 1614, 1585, 1538, 1493, 1445, 1374, 1298, 1243, 1180, 1117, 1069, 1013, 801, 767cm ^-1 ; ¹ HNMR (CDCl ₃ )δ 4.67 (2H, s, -OCH ₂ Ar), 5.01 (2H, s, -OCH ₂ Ar), 6.41 (1H, d, J = 2.2, 3-H), 6.61 (1H, dd, J = 2.2 and 8.5, 5-H), 6.86 (1H, d, J = 8.4, 6-H), 7.02 (1H, s, 4'-H of pyrazole), 7.23-7.40 (13H, m, -PhH and ArH ). MS (EI) m/e: 544 (M ⁺ +1), 125 (100).

1-phenyl-3-(2,4-bis(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid ethyl ester (8) and 1-phenyl-5-(2,4-bis (p-chlorobenzyloxy)phenyl)pyrazole-3-carboxylic acid ethyl ester (9)

Dissolve 1.5g (3mmol) 2,4-bis(p-chlorobenzyloxy)phenacylmethyl oxalate diethyl oxalate in 20ml HOAc, slowly drop into 0.4g (3.5mmol) phenylhydrazine, and reflux after dropping React for 10 hrs, cool to room temperature, pour the reaction solution into ice water, extract with 3×10 ml of chloroform, dry over anhydrous magnesium sulfate, distill off the chloroform, and perform flash column chromatography on the residue (eluent: petroleum ether: ethyl acetate=1: 1 (v/v)) Compound 8 was obtained as white needles 0.06g, yield 3.4%, mp: 121-122°C, ¹ HNMR (CDCl ₃ ) δ1.24 (3H, t, -CO ₂ CH ₂ CH ₃ ), 4.22 (2H, q, -CO ₂ CH ₂ CH ₃ ), 5.04 (2H, s, -OCH ₂ Ar), 5.13 (2H, s, -OCH ₂ Ar), 6.63-6.66 (2H, m, 3-H and 5-H), 7.36-7.49 (14H, m, -PhH, ArH and 4'-H of pyrazole), 8.01 (1H, d, J=8.8, 6-H). Compound 9 is a white needle Crystal about 0.7g, yield 39%, mp: 137-138°C, ¹ HNMR (CDCl ₃ ) δ 1.42 (3H, t, -CO ₂ CH ₂ CH ₃ ), 4.44 (2H, q, -CO ₂ CH ₂ CH ₃ ), 4.65 (2H, s, -OCH ₂ Ar), 4.98 (2H, s, -OCH ₂ Ar), 6.38 (1H, d, J=2.3, 3-H), 6.58 (1H, dd, J=2.3 and 8.4, 5-H), 6.83 (1H, d, J=8.4, 6-H), 6.95 (1H, s, 4'-H of pyrazole), 7.19-7.37 (13H, m, -PhH and ArH).

1-methyl-3-(2-benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid ethyl ester (11a) and 1-methyl-5-(2- Benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-3-carboxylic acid ethyl ester (12a)

Mix 0.8 g (1.7 mmol) ethyl 3-(2-benzyloxy-4-(p-chlorobenzyloxy)phenyl)pyrazole-5-carboxylate and 1 ml CH ₃ I, 3 g K ₂ CO ₃ Reflux reaction in 25ml of acetone for 2hr, cool to room temperature, filter, distill off the acetone, the residual flash column chromatography (eluent: petroleum ether: ethyl acetate = 4:1 (v/v)) to obtain compound 11a as white Crystal 0.38g, yield 46%, mp: 139-140°C, ¹ HNMR (CDCl ₃ ) δ 1.34 (3H, t, -CO ₂ CH ₂ CH ₃ ), 4.21 (3H, s, -NCH ₃ ), 4.30 (2H, q, -CO ₂ CH ₂ CH ₃ ), 5.03 (2H, s, -OCH ₂ -), 5.14 (2H, s, -OCH ₂ -), 6.63 (1H, dd, J=2.4 and 8.8 , 5-H), 6.64 (IH, d, J=2.4, 3-H), 7.29 (1H, s, 4'-H of pyrazole), 7.33-7.47 (9H, m, -PhH and ArH), 7.91 ( 1H, d, J=8.8, 6-H). Compound 12a is white crystal 0.34g, yield 41%, mp: 124-126°C. ¹ HNMR (CDCl ₃ ) δ1.41 (3H, t, -CO ₂ CH ₂ CH ₃ ), 3.76 (3H, s, -NCH ₃ ), 4.42 (2H, q, -CO ₂ CH ₂ CH ₃ ), 5.04 (2H, s, -OCH ₂ -), 5.05 (2H, s, -OCH ₂ -), 6.62 (1H, dd, J=2.4 and 8.4, 5-H), 6.68 (1H, d, J=2.3, 3-H), 6.77 (1H, s, 4'-H of pyrazole), 7.17 (1H, d, J=8.3, 6-H), 7.21-7.37 (9H, m, -PhH and ArH).

1-(4-chlorobenzyl)-3-(2-benzyloxy-4-(4-chlorobenzyloxy)phenyl)pyrazole-5-carboxylic acid ethyl ester (11b) and 1-(4- Chlorobenzyl)-5-(2-benzyloxy-4-(4-chlorobenzyloxy)phenyl)pyrazole-3-carboxylic acid ethyl ester (12b)

Referring to the preparation methods of compounds 11a and 12a, compound 11b was obtained as white needles with a yield of 89%, mp124-126°C, ¹ HNMR (CDCl ₃ ) δ1.28 (3H, t, -CO ₂ CH ₂ CH ₃ ), 4.24 (2H, q, -CO ₂ CH ₂ CH ₃ ), 5.04 (2H.s, -OCH ₂ -), 5.14 (2H, s, -OCH ₂ -), 5.74 (2H, s, 1-NCH ₂ Ar ), 6.62-6.65 (2H, m.3-H and 5-H), 7.23-7.46 (14H, m, -PhH, ArH and 4'-H of pyrazole), 7.97 (1H, d, J=8.3, 6-H). Compound 12b is white needle crystal, yield 9.6%, mp117-118℃, ¹ HNMR (CDCl ₃ ) δ1.41 (3H, t, -CO ₂ CH ₂ CH ₃ ), 4.43 (2H, q , -CO ₂ CH ₂ CH ₃ ), 4.95 (2H, s, -OCH ₂ -), 5.02 (2H, s, -OCH ₂ -), 5.23 (2H, s, -NCH ₂ Ar), 6.53 (1H, dd, J=2.4 and 8.4, 5-H), 6.62 (1H, d, J=2.3, 3-H), 6.79 (1H, s, 4'-H of pyrazole), 6.97 (1H, d, J= 8.4, 6-H), 6.81-7.33 (13H, m, -PhH and ArH).

Claims (10)

1. included compound and its esters of formula (I):

Wherein, R ₁Be C ₁-C ₆Alkyl or alkenyl, or have one to four carbon atom carboxylic acid or its ester class at interval, or have one to four carbon atom aryl or substituted aryl at interval, or have one to four carbon atom heterocycle at interval; R ₂, R ₃Be H or halogen independently of one another.

2. according to the described compound of claim 1, wherein R ₁Be C ₁-C ₄Alkyl, R ₂, R ₃Be 4-chlorine independently of one another.

3. according to the described compound of claim 2, wherein R ₁Be normal-butyl, R ₂, R ₃Be 4-chlorine independently of one another.

4. according to the described compound of claim 1, wherein R ₁Be one to two carbon atom carboxylic acid at interval, R ₂, R ₃Be 4-chlorine independently of one another.

5. according to the described compound of claim 4, wherein R ₁Be carboxymethyl, R ₂, R ₃Be 4-chlorine independently of one another.

6. pharmaceutical composition wherein contains general formula (I) compound and the pharmaceutical carrier of claim 1.

7. the general formula of claim 1 (I) the compound application that is used for preparing the medicine for the treatment of cardiovascular and cerebrovascular diseases as endothelin-receptor antagonists.

8. the general formula of claim 1 (I) compound is used for preparing the application of the medicine for the treatment of diabetes, ephrosis as endothelin-receptor antagonists.

9. the general formula of claim 1 (I) the compound application that is used for preparing the medicine for the treatment of asthma as endothelin-receptor antagonists.

10. the general formula of claim 1 (I) compound is used for preparing the application of the hyperfunction medicine of treatment Tiroidina as endothelin-receptor antagonists.

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