CN1401654A - 3,4'-5-trihydroxystlbenes compounds with functions of reducing pulmonary artery high pressure and improving respiration function - Google Patents

Abstract

A 3',4',5-trihydroxystlbene compound for lowering the high tension of arteria pulmonalis and improving respiratory function is disclosed. It includes polydatin (or piceid) and resveratnol prepared by hydrolyzing polydatin.

Description

Has the resvertrol compounds that reduces pulmonary hypertension, improves the respiratory function purposes

One, affiliated field

The invention belongs to pharmaceutical (A61p11/), relate to a kind of Polydatin compounds, particularly a kind of Polydatin compounds (the compound giant knotweed glucoside, that have the resvertrol class formation) of reducing pulmonary hypertension, improving respiratory function of being used to.

Two, background technology

In recent years owing to reasons such as environmental pollutions, pulmonary heart disease morbidity scope and hazardness are on the rise, and drug research that make development effectively regulate Ppa pulmonary artery pressure (hph), to prevent and treat pulmonary heart disease has been subjected to attention, and at present, relevant both at home and abroad report is a lot.Pharmacological agent is maximum with the research of medicament for expanding vascellum, but existing medicine clinical effectiveness is unsatisfactory, this be because: mostly medicine (except that sucking NO) that (1) existing to fall Ppa pulmonary artery pressure is to treat the medicine of essential hypertension, shortage is to pulmonary vascular specificity, reducing Ppa pulmonary artery pressure simultaneously, also reduce systemic blood pressure, caused hypotension.(2) medicine of majority reduction Ppa pulmonary artery pressures is bigger to the scale effect of ventilation/blood flow, in step-down simultaneously, does not have improvement because of ventilation and makes PaO ₂Reduce, increase the weight of anoxic, unfavorable to the pulmonary heart disease treatment on the contrary.(3), cardiac output is descended, even increase the weight of degree of heart failure to the negative effects of heart.(4) existing to improve the pulmonary hypertension drug effect limited, and Ppa pulmonary artery pressure can raise again after the drug withdrawal, can't individuation or use outside hospital.

The Polydatin that from Chinese medicine, extracts, can not only microcirculation improvement, antithrombotic, anti-oxidant, have more the expansion pulmonary artery, reduce the effect of pulmonary hypertension, and antibiotic, antibechic is still arranged, (Han Jiwu, " giant knotweed Advance on Pharmacological Activities " " Chinese medicine company " 2001.0820,10 (8): P55-56 rather passs the civil service examinations in effects such as the contraction of isolated tracheal that antihistamine is caused and calmness; Jin Chunhua, Zhao Kesen, Liu Jie, Huang Xuliang: " Polydatin is to myocardial cell's regulating effect " " Chinese pathology health magazine " 2001.0215,17 (2): P128-130; Jin Chunhua, Liu Jie, Huang Xuliang, Zhao Kesen: " Polydatin is to the influence of contractility of cardiac muscle cells " " Chinese Pharmacological Bulletin 2000.0820,16 (4): P400-402), these comprehensive actions solve the deficiency of present treatment pulmonary heart disease medicine preferably, make Polydatin improve pulmonary hypertension, prevent and treat and have good DEVELOPMENT PROSPECT aspect the pulmonary heart disease.The Technology of extracting Polydatin from the Chinese medicine giant knotweed is ripe, though report is arranged both at home and abroad, but it is developed to medicine carries out systematic study, especially estimate as yet from experimentation on animals and clinical study two aspects and do not appear in the newspapers, also do not retrieve the patent report that the Polydatin that extracts is used to prevent and treat aspects such as HPH, CCP from giant knotweed.

Three, summary of the invention

The objective of the invention is to develop a kind of resvertrol compounds of reducing pulmonary hypertension, improving respiratory function of being used to.Comprise:

Polydatin (Polydatin or Piceid), chemical name 3 ', 4,5 resveratrols-3-β-list-D-glycoside (Resveratnot-3-β-D-glucoside),

Molecular formula: C ₂₀H ₂₂O ₈

Molecular weight: 390

Physico-chemical property: white powder can be dissolved in ethanol, in acetone purple and the hot water fully.

Chemical structural formula:

Trans-resveratrol (Resveratnot), the chemical name resvertrol (3 ', 4 ', 5--tritydroxystlbene)

Molecular formula: C ₁₄H ₁₂O ₃

Molecular weight: 228.25

Chemical structural formula:

Physico-chemical property: white powder, can be dissolved in ethanol, acetone etc. fully, be insoluble in the water.

Polydatin is the natural effective monomer that extracts from the traditional Chinese medicine giant knotweed, and modern study proves that Polydatin can significantly be expanded animal coronary vasodilator, coronary blood flow increasing; The diastole bronchial smooth muscle improves ventilatory function.Also having lipopenicillinase, antibiotic, antithrombotic, improvement simultaneously organizes microcirculation, increases effect such as myocardial contraction.Applicant's discovering recently, giant knotweed crude extract can make the HPH of Hypoxic Pulmonary Hypertension in Rats animal model descend, and blood oxygen pressure and systemic arterial pressure are unaffected.Giant knotweed injection liquid (containing Polydatin 40mg) can obviously reduce the pulmonary hypertension that anoxic causes, and pulmonary circulation and body systemic vascular are had certain selectivity.As seen Polydatin is to the control HPH basis that has certain effect, thereby provides good choice for the treatment pulmonary hypertension.Trans-resveratrol is the glucoside unit of Polydatin, the hydrolysis Polydatin, usually can get trans-trans-resveratrol, according to structure activity relationship: the two all is a conjugated structure, has extremely strong Mulberry Extract effect, this also is one of its pharmacological mechanism, and in like manner the compound of homogeneous structure also has same drug effect.

Four, embodiment

The pharmacodynamic experiment and the acute toxinology experiment of this Polydatin compounds are elaborated the pharmaceutical use that Polydatin of the present invention has in order further to prove below in conjunction with the applicant.

According to traditional effect, the modern pharmacology effect of giant knotweed, selected the medicine pharmacological experimental method, and made preliminary pharmacodynamic experiment of Polydatin and acute toxinology experiment.

1. Polydatin is to the effect of pulmonary hypertension

1.1. the various dose Polydatin is to the influence of rat development of hypoxic pulmonary arterial pressure

1.1.1 method

1) preparation of Hypoxic Pulmonary Hypertension in Rats rat model and grouping experiment animal Wistar rat are 35, be male, body weight 200g～250g.Be divided into 7 groups at random, 5 of every components: 1. normal group; 2. control group (physiological saline 0.5Ml, abdominal injection); 3. treat A group (Polydatin 1.0mg/kg, 0.5ml abdominal injection); 4. treat B group (Polydatin 2.0mg/kg, 0.5ml abdominal injection); 5. treat C group (Polydatin 5.0mg/kg, 0.5ml abdominal injection); 6. treat D group (Polydatin 10.0mg/kg, 0.5ml abdominal injection); 7. treat E group (Polydatin 50.0mg/kg, 0.5ml abdominal injection).Except that normal group, all the other each groups all place in the self-control normal pressure low oxygen case, and oxygen concn is 10.0 ± 0.5 in the controlling box, CO ₂With anhydrous chlorides of rase calcium absorption water vapor, absorb CO in concentration＜1.5%, case with sodica calx ₂

2) Ppa pulmonary artery pressure mensuration and drug intervention rat are anaesthetized (33 mg/kg) with 1% intraperitoneal, with microtubular through the external jugular vein intubate to pulmonary artery, measure the mean pulmonary arterial pressure (mPAP) that is in 6 treated animals and normal group animal under the hypoxia respectively with PTM-7 type loadometer, except that normal group, all the other each groups are all carried out in the normal pressure low oxygen case.

3) the used data of statistical procedures are represented with X ± S.Relatively adopt the t check between group.

1.1.2 result

Table various dose Polydatin is to the influence of Hypoxic Pulmonary Hypertension in Rats rat mPAP (X ± S)

	Number of elements	???????????????mPAP
				Before	After
		Normal group treatment of control group A group treatment B group treatment C group treatment D group treatment E group	????5 ????5 ????5 ????5 ????5 ????5 ????5			????14.1±1.6 ????23.3±2.1 ????25.6±1.7 ????26.1±1.9 ????25.8±1.9 ????25.9±2.0 ????5.8±1.7	????14.3±1.6 ????23.8±2.2 ????18.5±2.1 ????17.3±1.8 ????14.4±2.3 ????13.8±2.1 ????13.5±1.9

By in the table as seen, give hypoxemia processing after, each treated animal Ppa pulmonary artery pressure is than normal group obviously raise (p＜0.01).After giving abdominal injection Polydatin 5min, the animal Ppa pulmonary artery pressure begins to descend, and behind the 30min, Ppa pulmonary artery pressure is in maintenance level.Have the effect (p＜0.05 and p＜0.01) of significant reduction Ppa pulmonary artery pressure by the visible Polydatin treatment group of table than control group, and be dose-dependently, reach optimum curative effect during to 10mg/kg.

1.1.3 conclusion

1) the Polydatin abdominal injection obviously reduces the pulmonary hypertension that hypoxemia causes, is dose-dependently.

2) the reduction pulmonary artery effect onset of Polydatin is very fast, and medication 5min begins to descend, and 30min reaches the maximum reducing amplitude.

1.2 Polydatin is to the influence of hypobaric hypoxia pulmonary hypertension rat

1.2.1 materials and methods

1) duplicating and 10 of grouping experiment animal Wistar rats of hypobaric hypoxia pulmonary hypertension rat model, be male, body weight 200～250g.Place in the hypobaric hypoxia storehouse, oxygen concn is (10.0 ± 0.5) % in the controlling box, CO ₂Concentration 1.5%, simulation height above sea level 5000m high empty condition, every day 8h, 6d, totally 4 weeks weekly.Animal model prepares successfully the back and divides 2 groups: 1. control group: 5, with the physiological saline intravenous injection; 2. treatment group: 5, give the Polydatin solution 10mg/kg intravenous injection of physiological saline dilution.

2) method press methods such as Sun Bo (Vender RL.chronc hypoxic pulmonaryhypertension:cellbiology to pathophysiology[J] chert, 1994,106:236) insert the floating microtubular in four chambeies to pulmonary artery through right external jugular vein, connect physiology polygraph (sino-america joint-venture Kaifeng south China Instr Ltd., CY-682D type polygraph), measure pulmonary arterial systolic pressure (SPAP).Measure body systolic arterial pressure (SABP) through the right femoral artery intubate.

3) the used data of statistical procedures are represented with X ± S.Group is asked and is relatively adopted the t check.

1.2.2 result

The table Polydatin is to the influence of Hypoxic Pulmonary Hypertension in Rats rat SPAP and SABP (X ± S)

Group	Number of elements	??SPAP(mm?Hg)	??SABP(mmHg)
				Control group	????5	????18±4	????110±23
The giant knotweed group	????5	????14±3	????115±20

Table has been summed up the influence of Polydatin to Hypoxic Pulmonary Hypertension in Rats rat SPAP and SABP.Relatively find that treatment group induced lung systolic arterial pressure obviously reduces than control group for two groups, have statistical significance (p＜0.05); The body systolic arterial pressure of two treated animals is no significant difference (p＜0.05) then.

1.2.3 conclusion

Preliminary observation proves that Polydatin (10mg/kg) intravenously administrable can obviously reduce the pulmonary arterial systolic pressure of hypoxemia low pressure Pulmonary Hypertension animal model, and the body arterial pressure is not had influence.

1.3 the various dose Polydatin is to the influence of rat hypoxic pulmonary arterial pressure

1.3.1 method

1) preparation of hypoxic pulmonary hypertension rat model and grouping experiment animal Wistar rat are 35, be male, body weight 200～250g.Be divided into 7 groups at random, 5 of every components: 1. normal group; 2. control group (physiological saline 0.5Ml, abdominal injection); 3. treat A group (Polydatin 1.0mg/kg, 0.5ml abdominal injection); 4. treat B group (Polydatin 2.0mg/kg, 0.5ml abdominal injection); 5. treat C group (Polydatin 5.0mg/kg, 0.5ml abdominal injection); 6. treat D group (Polydatin 10.0mg/kg, 0.5ml abdominal injection); 7. treat E group (Polydatin 50.0mg/kg, 0.5ml abdominal injection).Except that normal group, all the other each groups all place in the self-control normal pressure low oxygen case, and oxygen concn is 10.0 ± 0.5 in the controlling box, CO ₂With anhydrous chlorides of rase calcium absorption water vapor, absorb CO in concentration＜1.5%, case with sodica calx ₂

2) Ppa pulmonary artery pressure mensuration and drug intervention rat are anaesthetized (33mg/kg) with 1% intraperitoneal, with microtubular through the external jugular vein intubate to pulmonary artery, measure the mean pulmonary arterial pressure (mPAP) that is in 6 treated animals and normal group animal under the hypoxia respectively with PTM-7 type loadometer, except that normal group, all the other each groups are all carried out in the normal pressure low oxygen case.

3) the used data of statistical procedures are represented with X ± S.Relatively adopt the t check between group.

1.3.2 result

Table various dose Polydatin is to the influence of hypoxic pulmonary hypertension rat mPAP (X ± S)

Normal group	Control group	Treatment A group	Treatment B group	Treatment C group	Treatment D group	Treatment E group
							????mPAP	??14±1.6	?18.5±2.1	17.3±1.8	??15.8±1.7	??14.3±1.6	14.4±2.3
????P		?P＜0.01**	P＜0.01*	??P＜0.01*	??P＜0.01*	P＜0.01*

Annotate: * and control group compare * * and normal group compares

By in the table as seen, give hypoxemia processing after, each treated animal Ppa pulmonary artery pressure is than normal group obviously raise (P＜0.01).

After giving abdominal injection Polydatin 5min, the animal pulmonary artery begins to descend, and Ppa pulmonary artery pressure is in maintenance level behind the 30min.The effect (P＜0.05 and P＜0.01) that has significant reduction Ppa pulmonary artery pressure by visible Polydatin treatment group in the table.And be dose-dependently.

1.3.3 conclusion

1) the Polydatin abdominal injection obviously reduces the pulmonary hypertension that anoxic causes, is dose-dependently.

2) the reduction pulmonary artery effect onset of Polydatin is very fast, and medication 5min begins to descend, and 30min reaches the maximum reducing amplitude.

1.4 Polydatin reduces the mechanism of action of rat hypoxic pulmonary arterial pressure

1.4.1 experimental technique

1) preparation of hypoxic pulmonary hypertension rat model and grouping experiment animal Wistar rat are 20, cleaning level, be male, body weight 200g～250g.Adopt self-control normobaric hypoxia case, press Xue Shi (Xue Quanfu, Xie Jianming, Hu Changgui waits " foundation of normobaric hypoxia pulmonary hypertension model in rats " [J] " Chinese tuberculosis and breathing magazine ", 1989,12 (6): oxygen concn is 10.0 ± 0.5 in the 350) reported method, controlling box, CO ₂With anhydrous chlorides of rase calcium absorption water vapor, absorb CO in concentration＜1.5%, case with sodica calx ₂Every day 8h, 6d in totally 4 weeks, is copied into the pulmonary hypertension model in rats that is bordering on normobaric hypoxia weekly.After model prepares, be divided into 4 groups at random, 5 every group: 1. control group (physiological saline 0.5Ml, abdominal injection); 2. treat A group (Polydatin 5.0mg/kg, 0.5ml abdominal injection); 3. treat B group (Polydatin 10.0mg/kg, 0.5ml abdominal injection); 4. treat C group (Polydatin 20.0mg/kg, 0.5ml abdominal injection); Reagent: Polydatin medicinal powder (structure is confirmed, purity＞98%).TXB ₂, 6-Keto-PGF ₁α radioimmunity test kit (Zhong Shan company in Beijing provides).

1.4.2 Ppa pulmonary artery pressure is measured and the drug intervention rat is anaesthetized (33mg/kg) with 1% intraperitoneal, with microtubular through the external jugular vein intubate to pulmonary artery, with PTM-7 type loadometer respectively at medicine before, 5min, 30min, 60min and 120min measure the mean pulmonary arterial pressure (mPAP) of 4 treated animals and normal group animal after the medication, measured by radioimmunoassay TXB simultaneously takes a blood sample ₂, 6-Keto-PGF ₁α.

3) the used data of statistical procedures are represented with X ± S.Relatively adopt the t check between group.

1.4.3 result

See Table 1, table 2, table 3.

Table 1 various dose Polydatin is to the influence of hypoxic pulmonary hypertension rat mPAP (X ± S)

Group	Number of elements	?????????????????????MPAP(mmHg)
						????0min	????30min	????60min	????120min
		Treatment of control group A group treatment B group treatment C group	????5 ????5 ????5 ????5	??25.8±1.9 ??25.9±2.0 ??23.8±2.2 ??24.8±2.1	??23.3±2.1 ??14.8±1.7 ??14.3±1.6 ??13.5±1.9					??25.6±1.7 ??17.3±1.8 ??14.1±1.6 ??13.1±1.7	??26.1±1.9 ??18.5±2.1 ??14.4±2.3 ??14.3±2.1

Table 2 Polydatin is to TXB ₂Influence (X ± S)

Group

Number of elements

????TXB 2(ng/L)

		????0min	????30min	????60min	????120min
						Treatment of control group A group treatment B group treatment C group	????5 ????5 ????5 ????5	??35.8±3.9 ??35.6±2.0 ??33.8±2.2 ??34.8±2.1	??33.3±4.1 ??24.8±3.7 ??24.3±3.6 ??23.5±2.9	??35.6±3.7 ??19.3±2.8 ??15.1±3.0 ??16.1±2.7	??36.1±4.9 ??18.5±2.7 ??16.4±3.3 ??16.8±2.8

Table 3 Polydatin is to 6-Keto-PGF ₁The influence of α (X ± S)

Group	Number of elements	?????????????????????????6-Keto-PGF 1α(ng/L)
						????0min	????30min	????60min	????120min
		Treatment of control group A group treatment B group treatment C group	????5 ????5 ????5 ????5	??15.8±1.9 ??15.6±2.0 ??16.2±1.8 ??15.9±2.1	??13.3±4.1 ??24.8±3.7 ??24.6±3.5 ??23.2±3.9					??15.6±2.7 ??39.3±2.8 ??45.1±3.1 ??46.1±3.7	??16.2±1.9 ??33.5±2.7 ??36.8±3.7 ??36.7±2.9

By in the table 1 as seen, after giving hypoxemia and handling, each treated animal Ppa pulmonary artery pressure obviously raises than normal group.After giving abdominal injection Polydatin 5min, the animal pulmonary artery begins to descend (P＜0.05), and (P＜0.01) Ppa pulmonary artery pressure is in maintenance level behind the 30min.The effect that Polydatin reduces Ppa pulmonary artery pressure has dose-dependently.By in table 2, the table 3 as seen, after the anaerobic treatment, TXB in each treated animal blood ₂Obviously raise, and 6-Keto-PGF ₁α obviously reduces.After giving abdominal injection Polydatin 5min, TXB ₂Begin to descend (P＜0.05), be in maintenance level (P＜0.01) behind the 30min, and 6-Keto-PGF ₁α obviously raises (P＜0.05), is in maintenance level (P＜0.01) behind the 30min, and the variation of the two has dose-dependently.

1.4.4 conclusion

Polydatin may be by suppressing the TXA that anoxic causes ₂Generation and PGI ₂Reduction, thereby improved the vasoconstrictive degree of lung, reduced Ppa pulmonary artery pressure.

2. the Polydatin capsule cures mainly relevant Pharmacodynamic test of active extract with function

Test 1: the Polydatin capsule is to the thrombotic effect of rabbit

Test 2: the Polydatin capsule is to the effect of rabbit platelet aggregation

Test 3: the Polydatin capsule is to the effect of rat blood rheological

Test 4: the Polydatin capsule is to the microcirculatory effect of rat intestine cell membrane

2.1 test objective

For clinical application provides the pharmacology test foundation.

2.2 animal

1) kind, strain, source, conformity certification, New Zealand strain rabbit, the certification of fitness: the moving word 08-018 of Shan doctor; SD strain big white mouse conformity certification: the moving card of Shan doctor word 08-005; Animal and feed provide by Xi'an Jiaotong University Medical College's Experimental Animal Center.

2) body weight: big white mouse 205.35 ± 13.9g, 10 every group, rabbit 2.5kg-3.5kg, 6 every group.

3) sex: ♀ ♂ dual-purpose.

4) laboratory temperature, humidity: 18 ℃-22 ℃, relative humidity 55%-65%.

2.3 medicine

1) tried thing, Polydatin capsule, every gram contain crude drug amount 200g, and pale powder is provided lot number 011022 by light industry science and trade company of Hengchang.Be mixed with the suspension of desired concn with distilled water temporarily.

2) contrast medicine, TIANBAONING, the 250mg/ sheet, Kang Enbeisi limited-liability company in Zhejiang produces, lot number: 010613; Calcium dobesilate 0.5g/ sheet, the sharp monarch in Xi'an stock company produces lot number: 0108009-1; Epinephrine inj draws back the lattice distance, and lot number: 993160 is produced in the bright emerging pharmacy in Guangzhou; Heparin sodium injection, biochemical-pharmaceutical factory, Shanghai, lot number: 9950105.

3) reagent, Sodium Citrate, chemical reagent factory in Xi'an produces, lot number: 980602.

ADP Shanghai chemical reagent factory produces, lot number: 001011.

2.4 key instrument

1) platelet aggregation instrument, diligent generation Supreme Being company produces in Beijing, model: LG-PABER.

2) the rotary blood viscosity tester of LG-III type, Chinese Academy of Sciences's sensor technology is produced, and shear rate is respectively 200s ^-1, 40s ^-1

3) 3F-3 type high speed Eppendorf centrifuge, erythrocyte sedimentation rate hematocrit reading plate, wheel company of Huaxing produces.

4) reading microscope, Changchun No.1 optical Instrument Plant production.

5) platelet adhesion reaction instrument, Chinese Academy of Sciences's sensor technology is produced, model: 9103-B.

2.5 test 1: the Polydatin capsule is to the thrombotic effect of rabbit

2.5.1 method and result

1) test method: get 36 of healthy new zealand rabbits, ♀ ♂ dual-purpose, body weight 2.5kg-3.5kg is divided into 6 groups at random, 6 every group, adapts to laboratory environment and weighs after 5 days, numbering; 1. physiological saline group 2ml/kg; 2. calcium dobesilate sheet group 60mg/kg; 3. Polydatin small dose group 25mg/kg; 4. dosage group 50mg/kg in the Polydatin; 5. the Polydatin heavy dose is organized 100mg/kg; Above animal gastric infusion 2ml/kg, continuous 2 weeks, behind last administration 0.5h, anaesthetize with 2.5% vetanarcol ear rafter intravenous injection, lie on the back behind the 1ml/kg and be fixed on the rabbit operating table, separate right common carotid artery and left external jugular vein, with the polyethylene intubate of getting ready in advance (polyene bore 1mm, long 10mm, internal diameter 2mm, long 8mm, in three sections pipes, put into No. 4 surgical thread of a 10cm, with heparin-saline solution 50u/ml) be full of polyethylene tube, inject heparin 50u/kg and clamp tube wall, after an end of pipe inserts left external jugular vein, the other end of pipe is inserted right common carotid artery, and 5min open blood in operation back interrupts blood and takes out silk thread rapidly and weigh behind the open blood flow 15min, each administration group and physiological saline group compare, and T check result sees Table 1 between group.

Table 1 Polydatin capsule is to the thrombotic influence of rabbit (X ± S)

Group	Dosage (mg/kg)	Number of animals (only)	Wet weight of thrombus (mg)	Inhibiting rate (%)
					The heavy dose of group of dosage group Polydatin in the physiological saline group calcium dobesilate group Polydatin small dose group Polydatin	????60 ????25 ????50 ????100	????6 ????6 ????6 ????6 ????6	?44.33±8.55 ?28.83±6.01 **?36.83±4.83 ?30.50±5.50 *?30.17±7.81 *	????33.46 ????15.00 ????29.61 ????30.37

Administration group and physiological saline group compare: * P＜0.05 * * P＜0.01

2) test-results

The big or middle dosage group of Polydatin can obviously suppress the rabbit thrombosis, alleviates wet weight of thrombus (* P＜0.05), inhibiting rate 30.37%, and the heavy dose of group of Polydatin is similar to the effect of calcium dobesilate group.In the Polydatin, small dose group do not have evident difference.

2.6 test 2: the Polydatin capsule is to the effect of rabbit platelet aggregation

2.6.1 method and result

1) test method: get 36 of healthy new zealand rabbits, ♀ ♂ dual-purpose, body weight 3kg-4kg after the chamber of testing is fed 5 days, is divided into 6 groups at random, and 6 every group, 1. physiological saline group 2ml/kg; 2. calcium dobesilate group 60mg/kg; 3. Polydatin small dose group 25mg/kg; 4. dosage group 50mg/kg in the Polydatin; 5. the Polydatin heavy dose is organized 100mg/kg; Water is can't help in the 12h fasting before the above animal experiment, heart extracting blood 5ml under aseptic condition, add contain in the continuous 3.8% sodium citrate anticoagulant test tube (ratio 1: 9) gently mixing leave heart 10min 600 to draw supernatant liquor standby, this is PRP, it is standby to leave heart 10min absorption supernatant liquor with 3000 again, this is PPP, with PPP zeroing, again in PRP by 1: 10 adding ADP reagent (final concentration is 2 μ moi/L).

Continuous gastric infusion is 18 days behind the mensuration administration thromboblast aggregation normal value, and capacity 2ml/kg respectively organizes platelet aggregation by above method survey behind last administration 40min, and self compares before and after the administration, and T checks processing, the results are shown in Table 2.

Table 2 Polydatin capsule is to the effect of rabbit platelet aggregation (X ± S)

Group	Dosage (mg/kg)	Number of animals (only)	????PAg Tmax％		???????????????Pag Ymax％
							Before the administration	After the administration	Before the administration	After the administration
			The heavy dose of group of dosage group Polydatin in the physiological saline group calcium dobesilate group Polydatin small dose group Polydatin	????60 ????25 ????50 ????100	????6 ????6 ????6 ????6 ????6	??123.40±62.58 ??160.93±54.35 ??177.60±126.72 ??144.27±112.34 ??119.27±48.52					??140.10±71.20 ??118.43±85.83 ??172.60±46.98 ??131.77111.97 ??32.60±32.55 **	??27.28±10.6 ??43.40±24.53 ??42.60±33.33 ??27.18±11.23 ??27.55±10.97	??20.75±6.38 ??19.73±6.72 ??27.17±9.42 ??26.85±19.43 ??10.76±12.73 *

Before and after organizing administration, each self compares: * P＜0.05 * * P＜0.01

2) test-results: can obviously reduce rabbit platelet aggregation rate (Pag before and after the administration of the heavy dose of group of Polydatin _Vmax%) with gathering time (PAg _Tmax%), test-results illustrates that with calcium dobesilate similar (* P＜0.01) Polydatin has the effect of tangible reduction rabbit platelet aggregation.

2.7 test 3: the Polydatin capsule is to the effect of rat blood rheological

2.7.1 method and result

1) test method: get 60 of healthy SD strain rats, body weight 196.51 ± 10.28g adapts to laboratory environment and weighs after 5 days, numbering; Be divided into 6 groups at random, 10 every group, 1. intact animal group; 2. model group (SN) 5ml/kg; 3. TIANBAONING sheet group 250mg/kg; 4. Polydatin small dose group 36mg/kg; 5. dosage group 72mg/kg in the Polydatin; 6. the Polydatin heavy dose is organized 144mg/kg; Above animal, in continuous 2 weeks of gastric infusion of all the other animals except that 1. 2. organizing, capacity is 5ml/kg.Animal is got blood the day before yesterday, removes outside the intact animal group all the other animal subcutaneous injection Adr0.08ml/100g, one day 2 times, 4h at interval, injection back 2h immerses after 5min does blood stasis model in the frozen water fasting with rat and can't help water for the first time, and get blood 2.7ml with the rat broken end morning next day.After wherein 1.5ml blood injects the anticoagulant heparin pipe rapidly and shakes up, insert in 35 ℃ the constant water bath box standbyly, 1.2ml blood injects the Sodium Citrate anticoagulant tube rapidly, shakes up standby.

Get the 1ml heparin anti-coagulating and inject in the viscometer rotating cylinder, change high shear rate, low shear rate switch, write down high shear rate, low shear rate viscosity scale value (cut: 200s by height ^-1Low cutting: 40s ^-1) with its input computer, try to achieve high shear rate of whole blood and low shear rate viscosity number.

Get the smooth kapillary in bottom, insert in the heparin anti-coagulating of firm mixing, treat that blood enters 4/5 place in the kapillary, seal lower port, the lower end skewer is gone in the plasticine, stand vertically, and behind the timing 1h, with erythrocyte sedimentation rate value (wynn's method).

With above-mentioned capillary centrifuge tube, with the centrifugal 5min of 3F-3 type high speed Eppendorf centrifuge, rotating speed 12000rpm reads the pcv value with the hematocrit plate again.

Is water-bath 5min in 56 ± 1 ℃ the water-bath with the capillary centrifuge tube of measuring hematocrit in temperature, use the centrifugal 5min of same whizzer then, rotating speed still is 12000rpm, take out the back and measure blood plasma length and scleroproein raw footage in the capillary vessel, press routine formula and calculate fibrinogen content (thermal precipitator method) with reading microscope.

The blood of finishing whole blood viscosity is sucked back former anticoagulant tube, with the centrifugal blood plasma of 80-2 type whizzer, rotating speed 3000rpm, time 15min gets 1ml, cuts method of viscosity by above-mentioned survey whole blood height and surveys plasma viscosity.

The whole blood reducing viscosity is calculated according to high shear rate viscosity number of whole blood and pcv by computer, and software is provided by Sensitive Technology Co. of the Chinese Academy of Sciences.

Getting 1ml Sodium Citrate anticoagulation adding capacity is in the long-neck circular glass bottle of 5ml, be placed on the platelet adhesion reaction instrument rotating disk, take out behind the rotating speed rotation 15min with 5rpm, draw the platelet count that the ammonium oxalate thinner of the preceding and postrotational anticoagulation injection of 20 μ l rotation 1%0.4ml is measured rotation front and back blood respectively with quantitatively getting blood vessel again, be calculated as follows then:

More than the results are shown in Table 3 (attached).More than each measurement result learn processing by statistics, the T assay is as follows between group.

2) test result: 1. model group and normal group compare, and the high shear rate viscosity of whole blood, low shear rate viscosity, plasma viscosity, height are cut indexs such as reducing viscosity, Fibrinogen and all increased and highly significant (* * P＜0.01); Total number of blood platelet, thrombocyte viscosity rate, erythrocyte sedimentation rate, PBC hematocrit do not have noticeable change.Blood stasis model is duplicated successfully.

2. positive drug group and model group compare, and the high shear rate viscosity of whole blood, low shear rate viscosity, plasma viscosity, height are cut reducing viscosity, Fibrinogen reduces and significance (* * P＜0.01) is arranged; 5 change in 9 indexs, illustrate that positive drug is effective.

3. Polydatin administration group and model group compare, and the high shear rate viscosity of whole blood, low shear rate viscosity, plasma viscosity, the height of heavy dose of group cut reducing viscosity, 5 indexs of Fibrinogen all reduce, and highly significant (* * P＜0.01); All the other 4 indexs have no significant change.In, the Fibrinogen of small dose group, erythrocyte sedimentation rate change highly significant (* * P＜0.01); All the other indexs do not have obvious change.

Illustrate: the Polydatin capsule can reduce rat plasma viscosity, thereby improves the rat blood circulation.

2.8 test 4: the Polydatin capsule is to the microcirculatory effect of rat intestine cell membrane

2.8.1 method and result

1) get 50 of the SD strain rats of adult healthy, ♀ ♂ half and half, 10 every group, body weight 205.35 ± 13.9g is provided by Xi'an Jiaotong University Medical College experimentation on animals center.After water 12h is can't help in the experimental rat fasting, with 20% urethane intraperitoneal injection of anesthesia rat 0.4ml/100g, back of the body position is fixed on the synthetic glass operating table, belly wool is cut only, cut the otch that is about about 3cm-4cm with scalpel in a sidewall, pull out a mouthful duodenum 12 intubate and connect the preparation administration that resets behind the bundle, pull out one section empty chest loop more gently, be tiled in the special synthetic glass perfusion pond, with major part intestinal tube is fixed on the Xiao Chi side cork sheet, mesentery places transparent Xiao Chi, and perfusion Xiao Chi fills the 15ml Rockwell nutritive medium with 30 ℃, adopts the binocular inverted microscope to amplify 80 times and observes (micrometer is housed in the microlens).

Test is divided into 5 groups: 1. physiological saline group (NS) 5ml/kg; 2. calcium dobesilate group 80mg/kg; 3. Polydatin small dose group 40mg/kg; 4. dosage group 80mg/kg in the Polydatin; 5. the Polydatin heavy dose is organized 160mg/kg; After above animal is observed every normal index earlier, duodenal administration, capacity 5ml/kg observes every desired value once more behind the 20min, and administration front and back own control is adopted in test, and test-results is learned the T check by statistics and is handled, and the results are shown in Table 4.

2) test-results: the large, medium and small dosage group of Polydatin all can increase rat intestine cell membrane capillary vessel caliber, capillary vessel flow velocity, the result is with similar (* P＜0.05 of calcium dobesilate group, * P＜0.01), each dosage group of medicine does not have significantly effect to capillary vessel intersection number point.

Conclusion: each dosage group of Polydatin capsule has the microcirculatory effect of obvious promotion rat intestine cell membrane.

More than every experimental result show that the heavy dose of group of Polydatin capsule can alleviate the weight (* P＜0.05) of rabbit thrombus, inhibiting rate 30.37%; Can reduce rabbit platelet aggregation rate PAgV _Max% and pool time PAgT _Max% (* * P＜0.01); Large, medium and small each the dosage group of Polydatin is to the hemorheology of rat index, and the high shear rate viscosity of whole blood, low shear rate viscosity, plasma viscosity, height are cut reducing viscosity, 5 indexs of Fibrinogen all reduce, and highly significant (* * P＜0.01); All the other 4 indexs have no significant change.In the Polydatin capsule, small dose group all can increase capillary vessel caliber, capillary vessel flow velocity, effect highly significant (* P＜0.05, * * P＜0.01).Test-results shows: Polydatin has tangible thrombus dissolving, reduces plasma viscosity and improves sanguimotor effect.

The present invention detects through Northwest University, and the wave spectrum and the bibliographical information of its nucleus magnetic resonance of Polydatin sample that the applicant produces, infrared spectra, UV spectrum are in full accord, proves that Polydatin structure that the applicant extracts is in full accord with bibliographical information.The Polydatin sample of institute's extraction separation is analyzed by HPLC, is standard with U.S. Sigma company reference substance, working sample Polydatin content＞99%.

In sum, the following special effect of the present invention:

1, the Polydatin abdominal injection obviously reduces the pulmonary hypertension that anoxic causes, is dose-dependently.

2, the reduction Ppa pulmonary artery pressure effect onset of Polydatin is very fast, and medication 20mg/kg can make the animal Ppa pulmonary artery pressure descend, and 40mg/kg reaches the maximum reducing amplitude.

3. generation that Polydatin may be by suppressing the TXA2 that anoxic causes and the reduction of PGI2, thus the vasoconstrictive degree of lung improved, reduced Ppa pulmonary artery pressure.

4. the Polydatin gastric infusion can reduce hematoblastic aggregation, improves the rat blood rheological, promotes microcirculation, and heavy dose of group drug effect is suitable with calcium dobesilate.

5. oral administration, toxicity is little, belongs to security level.

Table 3 Polydatin capsule is to the effect of rat blood rheological (X ± S)

Model group and normal group compare: △ P＜0.05 △ △ P＜0.01

Medication group and model group compare: * P＜0.05 * * P＜0.01

Table 4 Polydatin capsule is to the microcirculatory effect of rat intestine cell membrane (X ± S)

Group	Dosage (mg/kg)	Number of animals (only)	Capillary vessel caliber (μ m/S)		Capillary vessel flow velocity (μ m/S)		Fixing vision area capillary vessel number (bar)		Fixedly vision area capillary vessel point of crossing (individual)
											Before the administration	After the administration	Before the administration	After the administration	Before the administration	After the administration	Before the administration	After the administration
			The heavy dose of group of dosage group Polydatin in the physiological saline group calcium dobesilate group Polydatin small dose group Polydatin	????5ml ????80 ????40 ????80 ????120	????10 ????10 ????10 ????10 ????10	??0.78±0.10 ??0.77±0.08 ??0.75±0.10 ??0.73±0.09 ??0.70±0.11	?0.78±0.10 ?1.04±0.16 **?0.87±0.17 ?0.91±0.17 **?0.90±0.21 *	?7.83±0.82 ?8.14±0.66 ?7.73±0.96 ?8.20±0.91 ?7.67±0.73	?7.96±0.89 ?10.14±1.99 **?9.52±2.13 *?9.82±1.23 **?9.87±1.75 *	?7.60±0.97 ?6.40±1.17 ?7.40±1.35 ?7.80±1.40 ?7.40±1.07									?7.60±1.11 ?8.50±1.78 **?8.10±1.66 ?8.70±1.95 ?8.30±1.83	?4.80±0.79 ?4.80±0.78 ?4.00±0.83 ?4.70±1.06 ?4.50±0.97	?4.90±0.87 ?5.70±1.42 ?4.70±0.95 ?5.10±0.88 ?5.30±1.16

Before and after organizing administration, each self compares: * P＜0.05 * * P＜0.01

Claims (1)

1. resvertrol compounds comprises:

Polydatin (Polydatin or Piceid), chemical name 3 ', 4,5 resveratrols-3-β-list-D-glycoside (Resveratnot-3-β-D-glucoside),

Molecular formula: C ₂₀H ₂₂O ₈

Molecular weight: 390

Physico-chemical property: white powder can be dissolved in ethanol, in acetone purple and the hot water fully;

Chemical structural formula:

Trans-resveratrol (Resveratnot), the chemical name resvertrol (3 ', 4 ', 5--tritydroxystlbene)

Molecular formula: C ₁₄H ₁₂O ₃

Molecular weight: 228.25

Chemical structural formula:

Physico-chemical property: white powder, can be dissolved in ethanol fully, acetone etc. are insoluble in the water;

It is characterized in that: this resvertrol compounds has the purposes that reduces pulmonary hypertension, improves respiratory function.