CN103044409A - Puerarin derivative or pharmaceutically acceptable salt and application thereof - Google Patents
Puerarin derivative or pharmaceutically acceptable salt and application thereof Download PDFInfo
- Publication number
- CN103044409A CN103044409A CN2012105162164A CN201210516216A CN103044409A CN 103044409 A CN103044409 A CN 103044409A CN 2012105162164 A CN2012105162164 A CN 2012105162164A CN 201210516216 A CN201210516216 A CN 201210516216A CN 103044409 A CN103044409 A CN 103044409A
- Authority
- CN
- China
- Prior art keywords
- puerarin
- gram
- synthetic
- acceptable salt
- acetone
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 0 CC1C=CC(C(C)=O)=*C1 Chemical compound CC1C=CC(C(C)=O)=*C1 0.000 description 4
Landscapes
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a puerarin derivative or pharmaceutically acceptable salt thereof. According to the defects of the prior art, the structure of puerarin is modified; the water solubility and lipid solubility of puerarin are improved by changing the size, shape and space structure of molecules, and meanwhile the pharmaceutical activity of puerarin is maintained; and the puerarin derivative or pharmaceutically acceptable salt thereof has a structure of the general formula I shown in the specification.
Description
Technical field
The invention belongs to field of medicine and chemical technology, be specifically related to puerarin derivatives and application thereof.
Background technology
Puerarin (puerarin) is a kind of flavonoid glycoside that extracts by in the sweet kudzu of legume pueraria lobata (pueraria lobata) (puerarin thomsonii) root.Just on the books in the medical science works " herbal classic " of Ancient Times in China, " typhoid fever miscellaneous diseases note " and " medical science voluminous dictionary " with the kudzuvine root for treating general diseases." day support food prison " of Japan and " herbal with song food " also put down in writing it and have " sweet light, nontoxic, heat extraction, quench the thirst, defaecation, relieve the effect of alcohol, nourishing the stomach " effect.The chemical ingredients that two pairs of roots of kudzu vine hold in the bavin field in nineteen fifty-nine Japan is studied, and isoflavonoid is the main effective constituent of the root of kudzu vine, and content is more in the root of kudzu vine, mainly contains puerarin, Gly and daizeol.Side's departure in domestic 1974 waits has successfully extracted puerarin.Be used for clinical through Ministry of Health's approval in 1993.
Puerarin chemical name: 4', 7-dihydroxyl-8-β-D-Glucose base isoflavones, as follows.
Puerarin can strengthen myocardial contraction, the protection myocardial cell; Vasodilation, reduce blood pressure, improve microcirculation; Protect erythrocytic deformability, strengthen the hemopoietic system function; Platelet aggregation-against increases fibrinolytic, reduces blood viscosity; Ephritis, ephrosis renal failure model all there is provide protection; Non-specific immunity, humoral immunization, cellular immunization there is obvious regulating effect; Can promote the lymphocyte transformation of normal people and tumour patient; Interferon system there is obvious stimulation and induces effect.Present clinically puerarin has been widely used in the treatment of cardiovascular system diseases, fundus oculi disease, sudden deafness, diabetic complication, acute alcoholism and tumour, in addition, add famotidine, amoxycillin and Ofloxacin duodenal ulcer with puerarin, more alone famotidine, amoxycillin and Ofloxacin, ulcer healing rate is significantly increased.So puerarin is a kind of very rising medicine.
Yet the solubleness of puerarin in water is lower, and the propylene glycol that the puerarin injection of at present clinical application mostly adds higher concentration has not only increased cost as solubility promoter, also because liquid viscosity is too large, causes trouble in the filtration to production.
Summary of the invention
Technical problem: the present invention is directed to the prior art deficiency, puerarin is carried out structural modification, improve the water-soluble and fat-soluble of puerarin by changing bulk of molecule, shape and space structure, the pharmaceutical activity of the puerarin that keeps simultaneously obtains a series of puerarin derivates or its pharmacy acceptable salt.
Technical scheme: puerarin derivatives of the present invention or its pharmacy acceptable salt have the structure of following formula I:
In the formula I:
R
1Represent H ,-CH
2CH
2N (C
2H
5)
2,
R
2Representative-CH
2CH
2N (C
2H
5)
2,
The present invention also provides the above-mentioned application of puerarin derivate in preparation Cardiovarscular medicine and serum regulating drug.
Puerarin is isoflavones carbon glycosides, and 7,4'-dihydroxyl is active group.Wherein 7 hydroxyls are because being subjected to the steric influence of 8 glycosyls, and activity is than a little less than the hydroxyl of 4' position.The present invention mainly concentrates on 7, the 4' position the structure of modification of puerarin.The present invention is to the following principle of structure of modification Main Basis of puerarin:
1. utilize principle of pro-drug that puerarin is carried out structure of modification
Medicine is transported in vivo must be by various barriers, and such as epidermis, respiratory tract and gastrointestinal tract mucosa, capillary wall, uriniferous tubules wall, hemato encephalic barrier etc., these barriers are the adventitia of various cells.Intracellular plastosome, microsome etc. also respectively have its adventitia, and these films are referred to as microbial film.Microbial film is take double-deck molecular lipid as support, and protein (comprising enzyme and acceptor) is adsorbed on the surface or embeds in the lipid layer, and its hydrophilic radical is all outside, and hydrophobic group is all inside.Medicine depends on physico-chemical property and the molecular structure thereof of medicine by biomembranous ability.Drug transport diffuses to blood and body fluid, and certain water-soluble (claiming again wetting ability or lipophobia) need be arranged, and the microbial film transhipment by lipid need have certain fat-soluble (claiming again lipotropy or hydrophobicity).It is certain water-soluble to that is to say that medicine should have, and certain fat water distribution coefficient is arranged again.In order to improve puerarin transport process in vivo; improve it by biomembranous ability; especially the ability by hemato encephalic barrier; strengthen it to the activity of cardiovascular and cerebrovascular; the present invention utilizes principle of pro-drug; use the methods such as acidylate, sulfonylation and alkylation that puerarin has been carried out structure of modification, obtained 14 preferred puerarin derivate I
1-I
11, I
13-I
14And I
16See Table 1.
2. utilize synergetical principle that puerarin is carried out structure of modification
(1) indolizine
Indolizine (indolizine) is the important nitrogen-containing heterocycle compound of a class, new research is found, have the compound of indolizine parent nucleus except three traditional action sites of tool, can also act on the L-calcium channel and form new binding site, thereby have the calcium ion retardation.The present invention is introducing the indolizine group in the puerarin structure, and in the hope of strengthening calcium antagonistic activity, chemical compounds I has been synthesized in design accordingly
12, see Table 1.
(2) NO donor
NO is as courier in the cell and effector molecule, and different physiological roles is also regulated in mediation, plays a very important role in the system such as cardiovascular, immune, neural.The present invention is at the pharmacophoric group of the potential NO donor of puerarin structure introducing tool activity, and in the hope of strengthening the activity of its cardiovascular aspect, chemical compounds I has been synthesized in design accordingly
15, see Table 1.
(3) nicotinic acid
Nicotinic acid is VITAMIN the most stable in the vitamin B complex, can be converted into rapidly niacinamide in vivo, then become the integral part of cozymase and codehydrogenase Ⅱ, participate in the formation of the synthetic and high-energy phosphate bond of glycolysis, lipid metabolism, pyruvic acid metabolism, the pentose of glucose in the body etc., have widely physiologically active.Nicotinic acid or at present known to unique all blood fat basal component that can make towards the compound of correct direction development, be the wide spectrum serum regulating drug, can be used for combined hyperlipidemia familial, the treatment of hypertriglyceridemia, low hdl mass formed by blood stasis and high plasma lipoprotein mass formed by blood stasis, prolonged application can reduce incidence or the mortality ratio of coronary heart disease, myocardial infarction and other cardiovascular and cerebrovascular diseases.In order to strengthen the activity of medicine, reduce side effect, normal and the some drugs of nicotinic acid share the raising drug effect in recent years, share such as nicotinic acid and acetylsalicylic acid, the rat plasma lipid there is regulating effect, nicotinic acid can suppress the hyperlipidaemia by high fat diet and Tnt-induced, and the impact that two medicines share total cholesterol then is better than alone nicotinic acid.And puerarin it is reported the effect that reduces serum cholesterol, and the present invention designs and synthesized chemical compounds I accordingly
17, as described in following table.
Above-claimed cpd can prepare by the following method:
Puerarin is dissolved in acetone, adds 2 gram salt of wormwood, be heated to backflow, the Tosyl chloride that then slowly drips with acetone solution (is summarized in acetyl halide compound R
2X, wherein R
2Representative-CH
2CH
2N (C
2H
5)
2,
The above-claimed cpd synthetic route is as follows:
Wherein, chemical compounds I
1, I
2, I
3, I
10, I
12, I
15And I
16Synthesis material can adopt that following route is synthetic to be obtained:
Beneficial effect: the present invention is to designing synthetic puerarin derivate I
1-I
17Carried out the research of a series of pharmacology pharmacodynamics, comprise the focal cerebral ischemia in rats screening, on research of vasoactive impact and estrogen activity etc., result of study shows that these 17 compounds all show the pharmacologically active of certain cardiovascular and cerebrovascular, wherein, I 3 and I 6 have stronger treating cerebral ischemia, and I 5 shows as the effect that certain enhancing is shunk with puerarin in multiple calcium; I 9 and I 14 are when existing simultaneously with NLA; The VSM contraction of inducing for Phe has restraining effect.When not adding NLA and Ind, the VSM that 5 couples of Phe of I induce shrinks restraining effect; I 16 effects are suitable with SNP, but security may be better than SNP, I 7, and I 8, I 11, I 12, I 14 estrogen activitys are better than Daidzin.
Embodiment
Employed term except as otherwise noted, generally has the implication that those of ordinary skills understand usually in the present invention.
Below in conjunction with specific embodiment and comparable data the present invention is described in further detail.Should be understood that these embodiment just in order to demonstrate the invention, but not limit the scope of the invention by any way.
In following examples, various processes and the method do not described in detail are ordinary methods as known in the art.
Adopt XY-4 binocular micro melting point apparatus to measure fusing point.Infrared spectra Nieolet Impact 410 type determination of infrared spectroscopy, the KBr compressing tablet.
1H-NMR spectrum ACF-300 (500) BRUK type nmr determination, TMS is interior mark.The MS spectrum is measured with HP1100 type mass spectrograph.Thin-layer chromatography (TLC) plate is self-control, adopts silica gel G F
254(Haiyang Chemical Plant, Qingdao's production) was placed in the moisture eliminator through 100 ~ 110 ℃ of activation and saves backup in 1 hour.Column chromatography adopts 100 ~ 200 order silica gel and silica gel H (Haiyang Chemical Plant, Qingdao's production), dry column-packing.Reagent is commercially available chemical pure or analytical pure product, except specifying, and not treated direct use.
Embodiment 14 ', the synthetic (I of 7-O-two (N, N dimethylamine base) ethyl puerarin
1)
(1) diethylin monochloroethane hydrochloride (I
1a) synthetic
The mixed solution of diethylin ethanol and benzene is chilled to below 10 ℃, drips the benzole soln of thionyl chloride, added in about 1 hour, then heated and stirred refluxed 3 hours, and cooling is filtered the cold acetone washing once, acetone and ether mixed solution (1:1) washed twice, drying under reduced pressure gets product.
(2) 4 ', the synthetic (I of 7-O-two (N, N dimethylamine base) ethyl puerarin
1)
1 gram puerarin is dissolved in acetone, adds with a small amount of water-soluble I
1a(0.64 gram) and a little KI, heating reflux reaction spends the night, and filters, and filtrate is concentrated, and column chromatography for separation gets product.
IR(KBr)cm
-1:3383,1620,1246
ESI-MS:615.2[M+H]
+
1HNMR(DMSO-d
6)ppm:8.41(s,1H,C
2-H),8.05(d,1H,C
5-H),7.40(d,2H,C
2′6′-H),7.16(d,1H,C
6-H),6.80(d,2H,C
3′5′-H),4.91(d,1H,C
1′′-H)。
Synthetic (the I of embodiment 24 '-O-acetamido-N-(3,4-dimethoxy)-β-styroyl puerarin
2)
(1) N-acetyl bromide-(3,4-dimethoxy)-β-phenylethylamine (I
2a) synthetic 10ml veratramin(e) be dissolved in methylene dichloride, add the 5ml pyridine, the 4.8ml bromoacetyl bromide with the dilution of 10ml methylene dichloride after, slowly be added drop-wise in the veratramin(e) under ice bath and induction stirring, drip off rear continuation reaction 3h, reaction solution washes with water to neutrality, the organic layer anhydrous magnesium sulfate drying, filter, filtrate is concentrated into just muddiness, puts refrigerator overnight, there is crystal to separate out, suction filtration, light yellow crystal 10g, productive rate 60%.mp106~108℃ IR(KBr)cm
-1:3245，3080，1634，1514，1260
1HNMR(CDCl
3)ppm:2.77(t,2H,-
CH 2 CH
2NH-),3.51(m,2H,-
CH 2 NH),3.87(s,3H,OCH
3),3.86(s,3H,OCH
3),3.84(s,2H,CH
2Br),6.53(brs,1H,NH),6.70~6.83(m,3H,Ar-H)。
(2) 4 '-O-acetamido-N-(3,4-dimethoxy)-β-styroyl puerarin (I
2) the 2g puerarin is dissolved in acetone, adds the 1.5g(I
2a), 3g salt of wormwood, a little potassiumiodide, heated and stirred refluxes, tlc analysis is followed the tracks of reaction, raw material point is basic disappear after stopped reaction, suction filtration, filtrate is concentrated, (chloroform: methyl alcohol=6:1) gets white powder 1.5g, productive rate 49% to column chromatography for separation.mp148~150℃.IR(KBr)cm
-1:3384,1621,1515,1263
ESI-MS:660.2[M+Na]
+
1HNMR(DMSO+CDCl
3)ppm:8.11(s,1H,C
2-H),8.07(d,2H,C
5-H),7.73(brs,1H,NH),7.32(d,2H,C
2′,6′-H),7.05(d,2H,C
6-H),6.78(d,2H,C
3′,5′-H),6.59~6.73(m,3H,Ar-H),4.98(d,1H,C
1′′-H),4.01~4.77(m,2H,-OCH
2-),2.65(t,2H,-CH
2-Ar)。
Synthetic (the I of embodiment 34 '-O-acetamido-N-β-styroyl puerarin
3)
(1) N-acetyl bromide-β-phenylethylamine (I
3a) synthetic
6.3g β-phenylethylamine is dissolved in methylene dichloride, adds the 5ml pyridine, after the 4.3ml bromoacetyl bromide dilutes with the 10ml methylene dichloride; under ice bath and induction stirring, slowly be added drop-wise in the veratramin(e); drip off rear continuation reaction 3h, reaction solution washes with water to neutrality, the organic layer anhydrous magnesium sulfate drying; filter; filtrate is concentrated into just muddiness, puts refrigerator overnight, has crystal to separate out; suction filtration gets light yellow crystal N-acetyl bromide-β-phenylethylamine 5g.
mp72~74℃。
IR(KBr)cm
-1:3250，3084，1651，1570，1196；
1HNMR(CDCl
3)ppm:7.19~7.34(m,5H,Ar-H),6.58(brs,1H,NH),3.82(s,2H,-CH
2Br),3.51~3.57(m,2H,-
CH 2 NH-),2.84(t,2H,-
CH 2 CH
2NH)。
(2) 4 '-synthetic (I of O-acetamido-N-β-styroyl puerarin
3)
Get the 2g puerarin and be dissolved in acetone; add 1.5gN-acetyl bromide-β-phenylethylamine; 3g salt of wormwood, a little potassiumiodide, heated and stirred refluxes; tlc analysis is followed the tracks of reaction; the basic rear stopped reaction that disappears of raw material point, suction filtration, filtrate is concentrated; column chromatography for separation (chloroform: methyl alcohol=6:1), get white powder 1.4g.mp178~180℃
IR(KBr)cm
-1:3371,1621,1266,1083；
ESI-MS:600.1[M+Na]
+；
1HNMR(DMSO+CDCl
3)ppm:8.21(s,1H,C
2-H),8.07(d,1H,C
5-H),7.76(d,1H,NH),7.33(d,2H,C
2′6′-H),7.15(d,1H,C
6-H),7.02~7.14(m,5H,Ar-H),6.77(d,2H,C
3′5′-H),4.94(d,1H,C
1′′-H),4.46~4.81(m,2H,-OCH
2-),2.68(t,2H,-CH
2-Ar)。
Embodiment 44 '-O-p-toluenesulfonyl puerarin (I
4) and 4 ', 7-O-two p-toluenesulfonyl puerarin (I
5) synthetic
2 gram puerarins are dissolved in acetone, add 2 gram salt of wormwood, be heated to backflow, then slowly drip the Tosyl chloride (1 gram) with acetone solution, drip off rear continuation back flow reaction to TLC without the raw material point, stopped reaction filters, filtrate is concentrated, (chloroform: methyl alcohol=10:1) obtains white powder I 40.9 grams, mp154 ~ 156 ℃ to the residue column chromatography for separation, I 50.4 gram, mp124 ~ 126 ℃.
Ⅰ
4:
IR(KBr)cm
-1:3376,1633,1174,1089；
ESI-MS:593.0[M+Na]
+；
1HNMR(DMSO+CDCl
3)ppm:8.16(s,1H,C
2-H),7.97(d,1H,C
5-H),7.39(d,2H,C
2′6′-H),7.26(d,1H,C
6-H),6.77(d,2H,C
3′5′-H),4.36(d,1H,C
1′′-H),2.40(s,3H,Ar-CH
3)。
Ⅰ
5:
IR(KBr)cm
-1:3434,1637,1376,1176,1092；
ESI-MS:747.0[M+Na]
+；
1HNMR(DMSO-d
6)ppm:8.56(s,1H,C
2-H),8.15(d,1H,C
5-H),7.77(d,2H,C
2′6′-H), 7.45(d,1H,C
6-H),7.09(d,2H,C
3′5′-H),4.30(d,1H,C
1′′-H),2.35(s,6H,Ar-CH
3)。
Embodiment 54 '-O-benzenesulfonyl puerarin (I
6) synthetic
2 gram puerarins are dissolved in acetone, add 2 gram salt of wormwood, be heated to backflow, then slowly drip the benzene sulfonyl chloride (0.62ml) with acetone diluted, drip off rear continuation back flow reaction to TLC without the raw material point, stopped reaction, filter, filtrate is concentrated, and the residue column chromatography for separation (chloroform: methyl alcohol=8:1), obtain white powder 1.4 gram, mp156 ~ 158 ℃;
IR(KBr)cm
-1:3394,1634,1375,1173,1091；
ESI-MS:579.0[M+Na]
+；
1HNMR(DMSO-d
6)ppm:8.44(s,1H,C
2-H),8.15(d,1H,C
5-H),7.40(d,2H,C
2′6′-H),7.69~8.13(m,5H,-SO
2-Ar-H),7.37(d,1H,C
6-H),6.81(d,2H,C
3′5′-H),4.30(d,1H,C
1′′-H)。
Embodiment 64 '-O-benzoyl puerarin (I
7) synthetic
2 gram puerarins are dissolved in acetone, add 2 gram salt of wormwood, ice bath, then slowly drip the Benzoyl chloride (0.56ml) with acetone diluted, drip off rear continuation reaction 3 hours, filter, filtrate is concentrated, residue column chromatography for separation (chloroform: methyl alcohol=8:1), obtain white powder 2.2 grams.mp176~178℃。
IR(KBr)cm
-1:3399,1633,1270,1084；
ESI-MS:559.1[M+K]
+；
1HNMR(DMSO+CDCl
3)ppm:8.31(s,1H,C
2-H),8.15(d,1H,C
5-H),7.66~8.28(m,5H,-CO-Ar-H),7.36(d,2H,C
2′6′-H),7.32(d,1H,C
6-H),6.77(d,2H,C
3′5′-H),4.94(d,1H,C
1′′-H)。
Embodiment 74 '-O-methoxycarbonyl base puerarin (I
8) and 4 ', 7-O-dimethoxy formyl radical puerarin (I
9) synthetic
1 gram puerarin is dissolved in acetone, adds 1 gram salt of wormwood, slowly drip the methoxy methyl acyl chlorides (0.38ml) with acetone diluted, stirring at room reaction 4 hours is filtered, and filtrate is concentrated, residue column chromatography for separation (chloroform: methyl alcohol=8:1), get I
80.5 gram, 170 ~ 172 ℃ of mp, I
90.25 gram, mp134 ~ 136 ℃.
Ⅰ8:
IR(KBr)cm
-1:3380,1630,1265,1063；
ESI-MS:513.2[M+K]
+；
1HNMR(DMSO+CDCl
3)ppm:8.05(s,1H,C
2-H),7.91(d,1H,C
5-H),7.31(d,2H,C
2’6’-H),6.89(d,1H,C
6-H),6.75(d,2H,C
3′5′-H),5.03(d,1H,C
1′′-H),3.37(d,3H,-CH
3)。
Ⅰ9:
IR(KBr)cm
-1:3417,1757,1634,1270；
ESI-MS:571.2[M+K]
+；
1HNMR(DMSO-d
6)ppm:8.04(s,1H,C
2-H),7.90(d,1H,C
5-H),7.31(d,2H,C
2′6′-H),6.88(d,1H,C
6-H),6.76(d,2H,C
3′5′-H),5.02(d,1H,C
1′′-H),3.36(d,3H,-CH
3),3.12(d,3H,-CH
3)。
Embodiment 84 '-O-is to chlorobenzene formacyl puerarin (I
10) synthetic
2 gram puerarins are dissolved in acetone, add 2 gram salt of wormwood, ice bath, then slowly drip the parachlorobenzoyl chloride (0.6ml) with acetone diluted, drip off rear continuation reaction 3 hours, filter, filtrate is concentrated, residue column chromatography for separation (chloroform: methyl alcohol=8:1), obtain white powder 0.8 gram.mp178~180℃。
IR(KBr)cm
-1:3384,1632,1268,1091；
ESI-MS:557.0[M+Na]
+；
1HNMR(DMSO-d
6)ppm:8.51(s,1H,C
2-H),8.16(d,1H,C
5-H),8.14(d,2H,Ar-H),7.68(d,2H,Ar-H),7.45(d,1H,C
6-H),7.40(d,2H,C
2′6′-H),6.82(d,2H,C
3′5′-H),4.87(d,1H,C
1′′-H)。
Embodiment 94 '-O-phenylacetyl puerarin (I
11) synthetic
1 gram puerarin is dissolved in acetone, adds 1 gram salt of wormwood, slowly drip the phenyllacetyl chloride (0.56ml) with acetone diluted, stirring at room reaction 5 hours is filtered, and filtrate is concentrated, column chromatography for separation (chloroform: methyl alcohol=8:1), obtain white powder 1.0 grams.mp148~150℃。
IR(KBr)cm
-1:3285,1629,1262；
ESI-MS:535.2[M+H]
+；
1HNMR(DMSO-d
6)ppm:8.03(s,1H,C
2-H),7.85(d,1H,C
5-H),7.31(d,2H,C
2′6′-H),6.96(d,1H,C
6-H),6.78~6.95(m,5H,Ar-H),6.76(d,2H,C
3′5′-H),5.01(d,1H,C
1′′-H)。
The formyl radical methylene radical puerarin (I of embodiment 10 4 '-O-(2-phenyl-indolizine-3-)
12) synthetic
(1) N-phenacyl-2-picoline drone salt (I
12a) synthetic
0.05mol substituted pyridines and 0.05mol ω-bromoacetophenone reflux 2 hours in the 200ml ethyl acetate, cooling, the leaching white solid washs with an amount of ethyl acetate, gets product.
(2) 2-phenyl indolizine (I
12b) synthetic
0.05mol(I
12a) be dissolved in 150ml water and the 80ml methylene dichloride, add 0.1molK
2CO
3, stirring at room 1 hour is divided and is got organic layer, washes once anhydrous MgSO
4Drying, concentrated, obtain light yellow crystallization (I
12b).
(3) 2-phenyl 3-acetyl bromide indolizine (I
12c) synthetic
0.01mol2-the phenyl indolizine with etc. the mole bromine acetyl bromide be dissolved in the 40ml dry toluene, drip the dry DMF of 3ml, stirring at room 1 hour, reaction solution is poured in the frozen water, use ethyl acetate extraction, saturated NaHCO is used in the organic phase merging
3Be washed till pH7, anhydrous MgSO
4Drying is concentrated into driedly, and the solid ethyl alcohol recrystallization gets light green crystallization (I
12c)
(4) 4 '-the formyl radical methylene radical puerarin (I of O-(2-phenyl-indolizine-3-)
12) synthetic 0.5 gram puerarin be dissolved in 200ml, add 0.375 gram (I 12c), 0.5 gram K
2CO
3, a little KI, heating reflux reaction 10 hours filters, and filtrate is concentrated, and column chromatography for separation gets light green solid 0.6 gram, mp184 ~ 186 ℃.
IR(KBr)cm
-1:3391,1621,1243,1082；
ESI-MS:672.2[M+Na]
+；
1HNMR(DMSO-d
6)ppm:8.33(s,1H,C
2-H),7.87(d,1H,C
5-H),7.11(d,1H,C
6-H),6.79(d,2H,C
3′5′-H),4.84(d,1H,C
1′′-H),4.64(s,2H,-CH
2-)。
Embodiment 11 4 '-O-ethoxycarbonyl methylene radical puerarin (I
13) synthetic
1 gram puerarin is dissolved in 200ml acetone, 0.5 gram K
2CO
3, a little KI, the 0.26ml ethyl chloroacetate, magnetic agitation, heating reflux reaction 5 hours filters, and filtrate is concentrated, and column chromatography for separation gets white solid 0.6 gram, mp122 ~ 124 ℃.
IR(KBr)cm
-1:3451,1621,1211,1059；
ESI-MS:525.1[M+Na]
+
1HNMR(DMSO-d
6)ppm:8.41(s,1H,C
2-H),8.05(d,1H,C
5-H),7.40(d,2H,C
2′6′-H),7.16(d,1H,C
6-H),6.80(d,2H,C
3′5′-H),4.91(d,1H,C
1′′-H),4.19(m,2H,-
CH 2 CH
3),1.24(t,3H,-CH
2 CH 3 )。
Embodiment 12 4 '-O-m-nitro alkylsulfonyl puerarin (I
14)
1 gram puerarin is dissolved in acetone, adds 1 gram salt of wormwood, 0.8 gram m-nitrobenzene sulfonyl chloride, room temperature induction stirring 5 hours is filtered, and filtrate is concentrated, residue column chromatography for separation (chloroform: methyl alcohol=8:1), obtain white powder 0.5 gram, 156 ~ 158 ℃ of mp.
IR(KBr)cm
-1:3425,1633,1353,1175；
ESI-MS:602.1[M+H]
+；
1HNMR(DMSO-d
6)ppm:8.60(s,1H,C
2-H),8.59(d,1H,C
5-H),7.94~8.38(m,4H,-SO
2-Ar-H),7.44(d,1H,C
6-H),7.34(d,2H,C
2′6′-H),6.77(d,2H,C
3′5′-H),4.88(d,1H,C
1′′-H)。
Embodiment 13 4 '-O-(4-phenyl-1,2, the methylene radical puerarin (I of 5-oxadiazole-2-oxide compound-3-)
15) synthetic
(1) 3-methylol-4-phenyl-1,2,5-oxadiazole-2-oxide compound (I
15a) synthetic
In three-necked bottle, add 10 gram (75mmol) cinnamic alcohols, 14.8ml Glacial acetic acid, machinery stirs and makes its dissolving under the room temperature, then drip saturated Sodium Nitrite (15 grams, 217mmol) the aqueous solution, keep temperature less than 70 ℃ in the dropping process, dripped off in about 1 hour, continue to stir 1 hour, there is orange to generate, stopped reaction with one times of saturated aqueous common salt dilution, is used extracted with diethyl ether 4 times with reaction solution, combining extraction liquid, wash three times with 10%NaOH solution, be washed to neutrality with saturated common salt again, organic layer adds anhydrous MgSO
4Drying is filtered, and filtrate is concentrated, and (sherwood oil: ethyl acetate=6:1), get orange obtains yellow solid 6.5 grams after freezing to the residue column chromatography for separation.mp63~65℃。
(2) 3-chloromethyl-4-phenyl-1,2,5-oxadiazole-2-oxide compound (I
15b) synthetic 6.5 gram I 15a(33.8mmol) be dissolved in the methylene dichloride that 60ml heavily steams, add 3ml(37.18mmol) anhydrous pyridine, cryosel is bathed, and adds thionyl chloride 2.73ml(37.18mmol), stirred overnight at room temperature.Reaction solution is washed with frozen water, then use saturated NaHCO
3Solution is washed till neutrality, and with saturated common salt washing one time, organic layer adds anhydrous MgSO again
4Drying is filtered, and filtrate is concentrated, gets reddish-brown oily matter, after freezing burgundy solid 5 grams.
(3) 4 '-O-(4-phenyl-1,2, the methylene radical puerarin (I of 5-oxadiazole-2-oxide compound-3-)
15) synthetic
0.5 the gram puerarin is used the 200ml acetone solution, adds 0.5 gram K
2CO
3, a little KI, 0.25 gram I 16b, induction stirring, heating reflux reaction, the TCL following response filters to without the raw material point, and filtrate is concentrated, column chromatography for separation (chloroform: methyl alcohol=5:1), get micro-yellow powder, mp218 ~ 220 ℃.
IR(KBr)cm
-1:3335,1620,1243；
ESI-MS:591.2[M+H]
+；
1HNMR(DMSO-d
6)ppm:8.38(s,1H,C
2-H),8.13(d,1H,C
5-H),7.85(d,2H,C
2′6′-H),7.40~7.63(m,5H,Ar-H),7.38(d,1H,C
6-H),6.82(d,2H,C
3′5′-H),5.26(d,1H,C
1′′-H)。
The adjacent chlorobenzene formacyl puerarin of embodiment 14 4 '-O-(I
16)
2 gram puerarins are dissolved in acetone, add 2 gram salt of wormwood, reflux 30 minutes, cooling slowly drips the o-chlorobenzoyl chloride (0.6ml) with acetone diluted under the room temperature, drip off rear continuation reaction 3 hours, filter, filtrate is concentrated, residue column chromatography for separation (chloroform: methyl alcohol=8:1), obtain white powder 0.8 gram.mp226~228℃。
IR(KBr)cm
-1:3298,1626,1253,1062；
ESI-MS:577.0[M+Na]
+；
1HNMR(DMSO-d
6)ppm:8.35(s,1H,C
2-H),7.88(d,1H,C
5-H),7.35~7.55(m,4H,Ar-H),7.33(d,2H,C
2′6′-H),6.94(d,1H,C
6-H),6.80(d,2H,C
3′5′-H),5.49(d,1H,C
1′′-H)。
Embodiment 15 4 '-O-(3-pyridine formyl radical) puerarin (I
17)
1 gram nicotinic acid (4.8mmol) with the dissolving of 20ml pyridine, adds 0.6 gram DCC(4.8mmol), 2 gram puerarins (4.8mmol), stirring at room 24 hours, the elimination insolubles, filtrate is poured in the large water gaging, chloroform extraction, merge the trichloromethane layer, concentrated, column chromatography for separation (chloroform: methyl alcohol=8:1), product, mp184 ~ 186 ℃.
IR(KBr)cm
-1:3378,1630,1272；
ESI-MS:520.2[M-H]
-；
1HNMR(DMSO-d
6)ppm:8.60(s,1H,C
2-H),8.37(d,1H,C5-H),7.94~8.32(m,4H,-Py-H),7.44(d,H,C6-H),7.34(d,2H,C
2′6′-H),6.77(d,2H,C
3′5′-H),4.28(d,1H,C
1′′-H)。
Embodiment 16 focal cerebral ischemias (MCAO) mouse model screening puerarin derivate
Adopt solidifying burning method to block a side arteria cerebri media and cause the focal cerebral ischemic in mice model, observe the prevention administration to the impact of Range of Cerebral Infarction (TTC staining) and neurimotor disturbance.Puerarin is control drug.During primary dcreening operation, select 3-4 animal for every group, found that two kinds of derivatives have and the similar treating cerebral ischemia of puerarin.These two kinds of derivatives are carried out multiple sieve, and every group of 10 animals are to determine its drug effect.
Find during primary dcreening operation (n=3-4) that derivatives I 3 and I 6 have the effect that reduces mouse left hemisphere cerebral infarction rate, reduce respectively 71% and 62% (see Table 2, Fig. 1 and Fig. 2).Determined the treating cerebral ischemia of these two kinds of derivatives when sieving again, their reduce the effect of mouse left hemisphere cerebral infarction rate, reduce respectively 42% and 39%(sees Table 3, Fig. 3 and Fig. 4).
The result: experimental result shows derivatives I
3And I
6Stronger treating cerebral ischemia (puerarin is contrast) is arranged.
Table 2. puerarin derivate is on the impact of the study of behaviour scoring of 24 hours Range of Cerebral Infarction of mouse brain middle cerebral artery occlusion and neurimotor disturbance.（n＝3-4,mean±sd.）
Compare with model group.**p<0.05,***p<0.01。
Table 3 puerarin derivate I
3And I
6Reduce the effect of mouse left hemisphere cerebral infarction rate.
Compare with model group.**p<0.05,***p<0.01。
Embodiment 17 pur and derivatives are to vasoactive influence
The stunning rat, the carotid artery bloodletting.Get rat chest aorta (TA), separate and adhere to surrounding tissue, be cut into the vascular circle of about 3mm, diameter 2-3mm.. vascular circle is hung on the 3mL that is full of KHS with the stainless steel needle acupuncture needle bathe in the ware, 37 ° of C of constant temperature regulate rest tension 1g(and transfer rest tension 2g when doing the vagusstoff diastole), balance 1 hour was changed one time K-H liquid in per 20 minutes, and with twice of NE pre-treatment.
The result
NLA is nitric oxide synthase inhibitors, and endotheliocyte NO capable of blocking is synthetic, and the contraction that Phe is induced has enhancement.Ind blocks COX-2, suppresses PGI
2Generate, also strengthen the contraction that Phe induces.Can find out I from following data
9And I
14When existing simultaneously with NLA, the VSM contraction of inducing for Phe has restraining effect.(table 4, Fig. 5,6) when not adding NLA and Ind, the VSM that 5 couples of Phe of I induce shrinks restraining effect, and 0.91 ± 0.31vs1.24 ± 0.30 (control) (n=13, p＜0.05) (Fig. 7).
Table 4The effects of Puerarin and its derivatives on contraction of VSM induced by Phe and interfered by NLA and NLA+Ind.(n=4)
| Control-Phe | Phe | NLA-Phe | NLA-Ind-Phe | |
| I1 | 1.37±0.27 | 1.45±0.32 | 1.46±0.34 | 1.44±0.31 |
| I2 | 1.69±0.33 | 1.51±0.60 | 1.73±0.55 | 1.55±0.67 |
| I3 | 1.49±0.43 | 1.46±0.62 | 1.68±0.28 | 1.72±0.41 |
| I4 | 1.37±0.28 | 1.42±0.32 | 1.38±0.47 | 1.41±0.53 |
| I5 | 1.47±0.39 | 1.40±0.07 | 1.41±0.58 | 1.39±0.61 |
| I6 | 1.53±0.37 | 1.31±0.28 | 1.43±0.68 | 1.47±0.73 |
| I7 | 1.28±0.23 | 1.02±0.34 | 1.15±0.54 | 1.34±0.76 |
| I8 | 1.23±0.09 | 0.92±0.19 | 0.94±0.33 | 0.99±0.49 |
| I9 | 1.19±0.25 | 0.89±0.31 | 0.83±0.26* | 0.91±0.47 |
| I11 | 0.79±0.49 | 0.68±0.52 | 0.77±0.53 | 0.91±0.63 |
| I12 | 1.01±0.35 | 0.94±0.38 | 1.18±0.53 | 1.10±0.50 |
| I14 | 0.92±0.33 | 0.86±0.45 | 0.69±0.42* | 0.90±0.67 |
| Puerarin | 1.57±0.71 | 1.75±0.20 | 1.54±0.73 | 1.80±0.87 |
| Control | 1.18±0.43 | 1.20±0.54 | 1.60±0.75 | 1.36±0.57 |
(*P<0.05vs.control)
The research of embodiment 18 puerarin derivate extension vessel activities
Medicine: puerarin derivate I
15, I
16
It is 10 that medicine is prepared final concentration with DMSO
-6Mother liquor, and become desired concn with distilled water. DMSO is diluted as stated above make solvent control simultaneously, positive drug is that SNP (Sodium Nitroprusside) directly becomes desired concn with distilled water.
Get SD (male) rat chest aorta ring 3mm, be hung on behind the endothelium-denuded and bathe in the ware, balance is after 1 hour, and NA processes twice, adds PHE (10
-6Final concentration) is contracted to the I that adds successively respectively each concentration behind the platform
15. I
16.C-1SNP. reach DMSO (10 to-6)
The result: experimental result shows that compound 16 is active suitable with SNP, but its security may be better than SNP.(seeing Fig. 8, table 5,6)
Table 5 puerarin derivate I
15, I
16Diastole effect to rat chest aorta.
Table 6 puerarin derivate I
15, I
16Diastole effect to rat chest aorta.
.#p<0.05vs DMSO.##p<0.01vs DMSO.
###p<0.001 vs DMSOn＝6
The screening of embodiment 19 puerarin derivate estrogen-like effectss
Experimental technique
The row ovariectomy is carried out vaginal smear after (1) 2% Sodital (0.2ML/10G) anesthesia after 5 days, will be in the dioestrus mouse and divide into groups and administration.Puerarin derivate, puerarin and Daidzin 200mg/kg subcutaneous injection, once a day, continuous three days.Progynon B 27 μ g/kg subcutaneous injections, once a day, continuous three days.Model group and normal group are to equal-volume solvent C MC-Na;
(2) second day begins vaginal smear continuous January after the administration;
(3) smear standards of grading
0 minute granulocyte accounts for the visual field＜10%
1 minute granulocyte accounts for the visual field 10 ~ 50%
2 minutes granulocytes account for the visual field 50%
3 minutes granulocytes account for the visual field〉50%
Granulocyte/epithelial cell was in 4 minutes〉90%
Granulocyte/epithelial cell was 50 ~ 90% in 5 minutes
Granulocyte/epithelial cell was 10 ~ 50% in 6 minutes
Granulocyte/epithelial cell was＜10% in 7 minutes
Only had epithelial cell in 8 minutes, and accounted for the visual field＜10%
Only had epithelial cell in 9 minutes, and accounted for the visual field 10 ~ 50%
Only had epithelial cell, and accounted for the visual field in 10 minutes 50%
(4) graphic representation is made in the smear scoring in Continuous Observation January, the compute area under curve, and make ratio with progynon B group area under curve, ratio is designated as specific activity.
Daidzin is food and the products and health products additive agent of using at present the weak estrogen activity of more tool as a result, can find out chemical compounds I from following data
7, I
8, I
11, I
12, I
14Estrogen activity be better than Daidzin, see Fig. 9, table 7.
Table 7 puerarin derivate estrogen activity
Embodiment 20 puerarin derivate Study on antioxidation of the present invention
According to the Fenton reaction principle
Fe
2++H
2O
2--→OH-+Fe
3+
FeSO is used in in-vitro simulated Fenton reaction
4With H
2O
2Reaction generates OH
-, incubate making oxidative stress model with 37 ℃ of temperature of liver homogenate, lipid film obviously damages, and the mark MDA showed increased of Peroxidation Product LPO is then observed after the adding pur and derivatives anti-oxidation stress drug effect in the parallel in vitro tests.
The result: detect according to the horizontal statistics of MDA, the result shows that Compound I 1 and I4 have stronger antioxygenation (seeing Figure 10, table 8).
Table 8 puerarin derivate of the present invention is to the restraining effect (inhibiting rate %) of lipid film damage
| pur 10 -3 | I1 10 -3 | I2 10 -3 | I3 10 -3 | I4 10 -3 | I5 10 -3 | I6 10 -3 | |
| 1 | 69.7 | 88.8 | 6.9 | 1.9 | 91.6 | 19.5 | 2.8 |
| 2 | 61.3 | 89.7 | 5.7 | 1.3 | 89.7 | 12.5 | 18.4 |
| 3 | 62.1 | 88.6 | 15.4 | 13.1 | 84.4 | 2.8 | 10.8 |
| 4 | 58.6 | 89.1 | 16 | 11.3 | 85.8 | 6.4 | 15.8 |
| 5 | 52 | 77.8 | 9.4 | 6.8 | 88.6 | 6.2 | 18.1 |
| 6 | 59.8 | 78.5 | 9.9 | 7.9 | 86.8 | 11.5 | 18.9 |
| Average | 60.58 | 85.4 | 10.55 | 7.05 | 87.81 | 9.81 | 14.1 |
| SD | 5.72 | 5.64 | 4.28 | 4.79 | 2.65 | 5.96 | 6.3 |
Claims (2)
1. a puerarin derivate or its pharmacy acceptable salt is characterized in that having the structure of following formula I:
Wherein:
R
1Represent H ,-CH
2CH
2N (C
2H
5)
2,
R
2Representative-CH
2CH
2N (C
2H
5)
2,
2. a puerarin derivate as claimed in claim 1 or its pharmacy acceptable salt application in preparation Cardiovarscular medicine and serum regulating drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012105162164A CN103044409A (en) | 2012-12-05 | 2012-12-05 | Puerarin derivative or pharmaceutically acceptable salt and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012105162164A CN103044409A (en) | 2012-12-05 | 2012-12-05 | Puerarin derivative or pharmaceutically acceptable salt and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103044409A true CN103044409A (en) | 2013-04-17 |
Family
ID=48057293
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012105162164A Pending CN103044409A (en) | 2012-12-05 | 2012-12-05 | Puerarin derivative or pharmaceutically acceptable salt and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103044409A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103382203A (en) * | 2013-08-15 | 2013-11-06 | 哈尔滨商业大学 | Puerarin derivative and synthetic method and application thereof |
| CN105601625A (en) * | 2015-11-20 | 2016-05-25 | 西安交通大学 | Preparation process for puerarin tramadol |
| CN109081834A (en) * | 2018-09-30 | 2018-12-25 | 余代英 | Puerarin derivate X with platelet aggregation-against function and preparation method thereof and application |
| CN109320552A (en) * | 2018-09-30 | 2019-02-12 | 余代英 | With active puerarin derivate of good biological and preparation method thereof and application |
| CN114790201A (en) * | 2022-05-11 | 2022-07-26 | 陕西中医药大学 | Vitexin derivative and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1080287A (en) * | 1991-11-06 | 1994-01-05 | 河南省医学科学研究所 | Kudzu vine root extract and preparation method and purposes |
| CN1566128A (en) * | 2003-07-03 | 2005-01-19 | 中国人民解放军军事医学科学院放射医学研究所 | Sugar charcoal glycoside isoflavone compound containing alkane amine group and alkoxy substitution, process and uses thereof |
| CN1823801A (en) * | 2005-02-25 | 2006-08-30 | 陕西镇坪制药厂 | Application of peracetyl pueraria in preparation of medicine for treating blood deficient cerebrovascular disease |
| WO2010045755A1 (en) * | 2008-10-24 | 2010-04-29 | 南京师范大学 | Glycosyl-derivatives of puerarin, pharmaceutical compositon, preparation method and use thereof |
-
2012
- 2012-12-05 CN CN2012105162164A patent/CN103044409A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1080287A (en) * | 1991-11-06 | 1994-01-05 | 河南省医学科学研究所 | Kudzu vine root extract and preparation method and purposes |
| CN1566128A (en) * | 2003-07-03 | 2005-01-19 | 中国人民解放军军事医学科学院放射医学研究所 | Sugar charcoal glycoside isoflavone compound containing alkane amine group and alkoxy substitution, process and uses thereof |
| CN1823801A (en) * | 2005-02-25 | 2006-08-30 | 陕西镇坪制药厂 | Application of peracetyl pueraria in preparation of medicine for treating blood deficient cerebrovascular disease |
| WO2010045755A1 (en) * | 2008-10-24 | 2010-04-29 | 南京师范大学 | Glycosyl-derivatives of puerarin, pharmaceutical compositon, preparation method and use thereof |
Non-Patent Citations (2)
| Title |
|---|
| 王劲，等: "葛根素衍生物的合成研究进展", 《山东中医药大学学报》, vol. 35, no. 4, 31 July 2011 (2011-07-31), pages 369 - 371 * |
| 王靖: "葛根素的结构改造和药理活性研究", 《中国药科大学硕士学位论文》, 31 December 2003 (2003-12-31) * |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103382203A (en) * | 2013-08-15 | 2013-11-06 | 哈尔滨商业大学 | Puerarin derivative and synthetic method and application thereof |
| CN103382203B (en) * | 2013-08-15 | 2016-02-03 | 哈尔滨商业大学 | Puerarin derivate and synthetic method thereof and application |
| CN105601625A (en) * | 2015-11-20 | 2016-05-25 | 西安交通大学 | Preparation process for puerarin tramadol |
| CN109081834A (en) * | 2018-09-30 | 2018-12-25 | 余代英 | Puerarin derivate X with platelet aggregation-against function and preparation method thereof and application |
| CN109320552A (en) * | 2018-09-30 | 2019-02-12 | 余代英 | With active puerarin derivate of good biological and preparation method thereof and application |
| CN109320552B (en) * | 2018-09-30 | 2020-10-16 | 浙江药苑生物科技有限公司 | Puerarin derivative with good biological activity and preparation method and application thereof |
| CN114790201A (en) * | 2022-05-11 | 2022-07-26 | 陕西中医药大学 | Vitexin derivative and preparation method and application thereof |
| CN114790201B (en) * | 2022-05-11 | 2023-11-10 | 陕西中医药大学 | Vitexin derivative and preparation method and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103044409A (en) | Puerarin derivative or pharmaceutically acceptable salt and application thereof | |
| CN106916177B (en) | A kind of deuterated dipeptide boronic acid or its ester type compound and its synthetic method and purposes | |
| CN106632383B (en) | Curcuma zedoary 01 derivatives and preparation method thereof and the application in antitumor drug | |
| CN103524564B (en) | Platinum (II) the title complex synthetic method of the different aporphine of a kind of 6-amino group and application | |
| CN104812761B (en) | Double B-carboline alkaloid compounds, its preparation method and its pharmaceutical composition and purposes | |
| CN108822126B (en) | Thienopyran formyl piperazine compound and medical application thereof | |
| CN104523664A (en) | Curcumin antineoplastic drug and application thereof | |
| CN104327097B (en) | Triazole derivatives of rapamycin and application | |
| CN105566271A (en) | Biflavone compound and application thereof to preparation of medicine for treating cancer | |
| JPH1067656A (en) | Cell adhesion suppressant | |
| CN107141284B (en) | Coptisine analog derivative, preparation method, pharmaceutical composition and anticancer usage | |
| CN108017656A (en) | Camptothecin derivative and its application in antitumor drug is prepared | |
| CN106565718B (en) | Selenium-containing compound and application thereof | |
| CN104761610A (en) | Novel alpha-hederin derivative and preparation method and use thereof | |
| CN102260239B (en) | Kutkin derivatives, and preparation and application thereof | |
| CN101974016A (en) | Amide compound and preparation method and applications thereof | |
| CN104530176A (en) | GAOH derivative and medical application thereof | |
| CN109467560A (en) | A kind of synthesis and application thereof | |
| CN108484635B (en) | 4H-thieno [2,3-b ] thiopyran-4-one compound and application thereof | |
| KR100490799B1 (en) | Food comprising an extract of bambusoideae plant or tricin isolated therefrom | |
| CN109734768A (en) | Go acetyl cedilanid glucosyl group modified compound composite lipidosome and its application | |
| CN105949117B (en) | Sorafenib derivative of the similar structures containing chalcone and its preparation method and application | |
| EP1155021A1 (en) | Hesperitin pro-forms with enhanced bioavailability | |
| CN104230931A (en) | Sophoridine derivatives, and preparation method and application thereof | |
| CN107311991B (en) | Puerarin derivative, its preparation method and application in preventing and treating cardiovascular and cerebrovascular diseases or diabetes and its complications |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C05 | Deemed withdrawal (patent law before 1993) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20130417 |