CN101502585B - Chinese red pepper plant extract as well as preparation method and application thereof - Google Patents
Chinese red pepper plant extract as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention provides a Zanthoxylum L extract, a preparation method and a use thereof. The Zanthoxylum L. plant extract is prepared by extracting from peel of Zanthoxylum L. plant and refining, the content of total polyphenols is 50-80 percent, wherein, the content of proanthocyanidin polymer ZP-CT-A is 5-15 percent. The Zanthoxylum L. plant extract can be combined with pharmaceutically acceptable carriers or other appropriate excipients to be prepared into formulations which are applicable to oral administration, injection and the like according to the conventional method and can be used for preventing and treating stroke, Parkinson's disease, hypertension, hyperlipidemia, atherosclerosis, coronary heart disease and other cardiovascular and cerebrovascular diseases.
Description
Technical field
The present invention relates to the xanthoxylum extract, and its production and use, belong to field of traditional Chinese medicine pharmacy.
Background technology
Zanthoxylum (Zanthoxylum L.) is shrub or the dungarunga of Rutaceae (Rutaceae) plant, about 250 kinds of the whole world, about 45 kinds of China, 13 mutation, in the most of area of China distribution is arranged all, its fruit, root, stem, leaf are all done medicinal at home and abroad, have effects such as analgesia, anesthesia, antibacterial, parasite killing, antitumor.Book on Chinese herbal medicine record peel is " green pepper is red ", the effect of tool warming spleen and stomach for dispelling cold, antalgesic-antipruritic, parasite killing dampness; Seed claims " Semen zanthoxyli ", the relieving asthma effect of diuretic of tool.The principal item of China's xanthoxylum has Pericarpium Zanthoxyli (Zanthoxylum bungeanum Maxim.), green pepper (Z.schinifolium Sieb.et Zucc.), Fructus Zanthoxyli Plansipini (Z.armatum DC), pericarpium Zanthoxyli (Z.simulans Hance), reins in ﹠-Floor (Z.avicennae (Lam.) DC.), Folium toonae sinensis Pericarpium Zanthoxyli (Z.ailantholides Sieb.et Zucc.), a green pepper (Z.molle Rehd), Radix Zanthoxyli (Z.nitidum (Roxb.) DC.) etc.The Chinese medicine Pericarpium Zanthoxyli that Chinese Pharmacopoeia version in 2005 is recorded is the dry mature skin of green pepper (Z.schinifolium Sieb.et Zucc.) or Pericarpium Zanthoxyli (Z.bungeanum Maxim.), and the mature fruit of gathering autumn is dried, and removes seed and impurity.Its function cures mainly and is warming middle-JIAO to relieve pain, and killing parasites for relieving itching is used for coldness and pain in the epigastrium, and vomiting is had loose bowels, abdominal pain due to worm stagnation, ascariasis; The eczema pruritus is controlled in external.
The tradition Pericarpium Zanthoxyli is widely used in food industry as food dressing, has developed the deep processed product of these classes such as xanthoxylum oleoresin, xanthoxylum oleoresin microcapsule, Pericarpium Zanthoxyli flavored oils, Pericarpium Zanthoxyli essence based on liposoluble constituent.People have done a lot of researchs to the liposoluble constituent and the pharmacotoxicological effect aspect thereof of the peel of xanthoxylum, these chemical constituents mainly contain (Sun Xiaowen such as volatile oil, alkaloid, amide, coumarin and fatty acid, Duan Zhixing, Zanthoxylum medicinal plants study progress, Acta Pharmaceutica Sinica, 1996,31 (3): 231-240), the drug effect of Zanthoxylum mainly contains four aspects: (1) is to neural anesthesia and analgesic activity: use Pericarpium Zanthoxyli ether extract or the Pericarpium Zanthoxyli volatile oil soothing agent as the department of stomatology clinically, replace Oleum Caryophylli that dental caries is carried out anti-inflammatory analgetic.Local trouble can produce analgesic effect, contain .beta.-fagarine in the Pericarpium Zanthoxyli and may be one of its analgesic active component (Chang Zhiqing, Wang Shuling, Hao Changyuan. the analgesia of .beta.-fagarine, spasmolytic and sedation [J]. Acta Pharmacologica Sinica, 1982,3:163 (66) .).(2) to digestive system disease: Pericarpium Zanthoxyli can be treated multiple insufficiency of stomach sympotoms caused by cold factors, also can treat gastric ulcer, hepatic injury, struvite and gastrointestinal function irregularity stomachache.(3) active anticancer: the effect that wherein mainly is alkaloid or volatile ingredient Japan pepper essential oil performance in the Pericarpium Zanthoxyli, as: fagaronine improves increase in life span (the Barret Y of leukemia mouse, Han Shuping. fagaronine-a kind of new leukemia alkaloid [J]. external medicine: plant amedica fascicle, 1993,8 (1): 19-21); The thick oil of Pericarpium Zanthoxyli all has obvious suppression effect (Mun S I to greasy peroxidization and carbon tetrachloride to the destruction of mouse liver, Ryu H S, Choi J S, Inhibitioneffects of Zanthoxylum schinifolium and its active principle on lipid peroxidationand liver damage in carbon tetrachloride-treated mice[J], Han ' guk Sikp ' umYongyang Kwahak Hoechi (Korean), 1997,26 (5): 943-951) tonkabean have antiplatelet aggregation effect (active component of Pericarpium Zanthoxyli and applied research. Food Additives Used in China, 2004; (2): 75-78; Chemical Constituents and Biological Activities of the Fruit of Zanthoxylumintegrifoliolum.J.Nat.Prod., 1999; 62:833-837).For the chemical constituent and the pharmacological action of water-soluble portion in the xanthoxylum (account for total extract 70%), modern study is less.Japan scholar Kusuda M. etc. is separated to 10 kinds of polyphenol compounds from Pericarpium Zanthoxyli (Z.piperitum) and (adopts the Pericarpium Zanthoxyli methanolic extract by the HP-20 column chromatography, use 0% successively then, 20%, 40%, 60%, 80%, 100% water-methanol solution gradient eluting, different eluting parts is further carried out separation and purification and is obtained 10 kinds of chemical compounds), find that wherein procyanidin polymer ZP-CT-A has antibacterial activity (Polyphenolic Constituent Structures ofZanthoxylum piperitum Fruit and the Antibacterial Effects of Its PolymericProcyanidin on Methicillin-Resistant Staphylococcus aureus.Biosci.Biotechnol.Biochem., 2006 to methicillin-resistant staphylococcus aureus (MRSA); 70 (6): 1423-1431).
Summary of the invention
Technical scheme of the present invention has provided a kind of xanthoxylum extract, and another technical scheme of the present invention provides the preparation method and the purposes of this plant extract.
The invention provides a kind of xanthoxylum extract, the total polyphenols percentage by weight is 50～80% in this extract.Further preferably, the total polyphenols percentage by weight is 65%～80%.
Wherein, contain procyanidin polymer ZP-CT-A in the total polyphenols, wherein, the weight percentage of procyanidin polymer ZP-CT-A in extract is 5%～15%.Further preferably, the weight percentage of described procyanidin polymer ZP-CT-A in extract is 8%～15%.
Further, also contain 3-oxygen-caffeoylquinic acids (chemical compound 2) in the described xanthoxylum extract, 4-oxygen-caffeoylquinic acids (chemical compound 3), procyanidin B1 (chemical compound 4), one or more in the hyperin (chemical compound 5).
Wherein, described xanthoxylum extract is from Pericarpium Zanthoxyli Zanthoxylum bungeanumMaxim., green pepper Z.schinifolium Sieb.et Zucc. Fructus Zanthoxyli Plansipini Z.armatum DC, pericarpium Zanthoxyli Z.simulans Hance, reins in ﹠-Floor Z.avicennae (Lam.) DC., Folium toonae sinensis Pericarpium Zanthoxyli Z.ailantholides Sieb.et Zucc), extract the peel of a green pepper Z.molle Rehd or Radix Zanthoxyli Z.nitidum (Roxb.) DC. etc. and obtain.Preferably, from the Chinese medicine Pericarpium Zanthoxyli, extract acquisition.
The present invention also provides the xanthoxylum preparation method of extract, may further comprise the steps:
(1) extract: with peel water or the aquiferous ethanol or the aqueous acetone extraction of xanthoxylum, raw material and extracting solution by weight/volume are 1: 5～20;
(2) concentrate: the extracting solution that step (1) is obtained does not extremely have the alcohol flavor or does not have the acetone flavor through vacuum concentration;
(3) macroporous adsorbent resin or polyamide absorption, eluting: the concentrated solution that step (2) is obtained adds an amount of water-soluble diffusing back upward macroporous adsorptive resins or polyamide, elder generation's water eluting, use 20%-95% ethanol or acetone eluting then, the eluant consumption is that 5-10 doubly measures the resin column bed volume, collect eluent, concentrate;
(4) precipitate with ethanol: the medicinal liquid that step (3) is obtained precipitates with 95% ethanol that 3-5 doubly measures, and abandons precipitation, gets supernatant;
(5) concentrated, dry: step (4) gained supernatant promptly gets the xanthoxylum extract through concentrated, dry.
Wherein aquiferous ethanol described in the step (1) or aqueous acetone concentration are 20-90%, preferred acetone; Extracting mode is dipping, percolation or backflow; When selecting the reflux, extract, mode for use, the effect during with 70% acetone extraction 3 times is the best;
Macroporous adsorbent resin in the step (3) is selected from D series, HPD series, HP series, XAD series or AB-8 type macroporous adsorbent resin.Preferably: the macroporous adsorbent resin of D series is the D-101 type; The macroporous adsorbent resin of HPD series is the HPD-500 type; The macroporous adsorbent resin of HP series is the HP-20 type; The macroporous adsorbent resin of XAD series is the XAD-4 type.
The xanthoxylum extract is further through Diaion HP-20, Toyopearl HW-40C, and MCI gelCHP-20P, column chromatographies such as ODS obtain five chemical compounds respectively, through chemistry and spectral method mensuration, to parse each compound structure as follows:
The xanthoxylum extract is an xanthoxylum peel water-soluble phenolic mixture of ingredients, and its total polyphenols content is 50～80%, and the content of wherein a kind of procyanidin polymer ZP-CT-A (chemical compound 1) is 5～15%.
The Pericarpium Zanthoxyli polyphenol is a water soluble ingredient, and it is more more suitable than making extractant with methanol that water of the present invention or aquiferous ethanol are made extractant, and assay is the result show, polyphenol content is apparently higher than methanolic extract in water or the aquiferous ethanol extract.
Its pharmacologically active power of the procyanidin of different polymerization degree is also different, for example there are some researches show, dimeric antioxidant activity is than active strong (the Oxygen free radical scavengercapacity in aqueous models of different procyanidins from grape seeds.J.Agric.Food Chem. of catechin, 1991,39:1549-1552), it increases (Free radical scavenging activity of grape seed extractand antioxidants by electron spin resonance spectrometry in anH to oxygen radical removing ability with the degree of polymerization in the degree of polymerization 2-5 scope
2O
2/ NaOH/DMSO system.Agric.Food Chem., 1999,47:2544-2548).
Further, the invention provides this xanthoxylum extract the preparation nerve cell-protective agents, resist myocardial ischemia or cerebral ischemia class medicine, anticoagulant, neutral endopeptidase inhibitor and lipidemia class medicine in application.
The present invention also provides a kind of pharmaceutical composition, is that the xanthoxylum extract by effective dose is an active component, adds the medicament that acceptable accessories or complementary composition are prepared from.
Wherein said medicament comprises oral formulations and injection preparation, and oral formulations comprises tablet, capsule, soft capsule, granule, drop pill etc., and injection preparation comprises injection, powder ampoule agent for injection etc.
The interior results of pharmacodynamic test of external or body shows; xanthoxylum extract energy neuroprotective cell of the present invention avoids anoxia-induced apoptosis and 6 hydroxy dopamine cause damage; the protection vascular endothelial cell is avoided anoxia-induced apoptosis; suppress the neutral endopeptidase activity, raise HDL receptor SR-B I expression, and have blood coagulation resisting function.
Neurocyte and the agent of vascular endothelial cell hypoxia protection can be used for the treatment of cerebral ischemia and myocardial ischemia, and 6 hydroxy dopamine cause the treatment that the neural cell injury protective agent can be used for parkinson.Neutral endopeptidase is the important target spot of just finding in recent years of hypertension therapeutic, studies show that, the neutral endopeptidase inhibitor can be treated hypertension effectively; HDL receptor SR-B I is an important target spot relevant with hyperlipidemia and atherosclerosis, and existing studies show that promotes HDL receptor SR-BI to express and can prevent and treat hyperlipidemia and atherosclerosis.Anticoagulative substance can be used for the treatment of multiple cardiovascular and cerebrovascular disease, as apoplexy, atherosclerosis and coronary heart disease etc.
The specific embodiment of form by the following examples, foregoing invention content of the present invention is described in further details, but should not be construed as summary of the invention of the present invention and only limit to following examples, all inventions of making based on foregoing of the present invention all belong to scope of the present invention.
Description of drawings
Fig. 1 xanthoxylum extract is to the anoxybiotic protective effect of vascular endothelial cell physical property
Figure 1A wherein: 5 groups of normal group, Figure 1B model group, 1 group of Fig. 1 C embodiment, 2 groups of Fig. 1 D embodiment, 3 groups of Fig. 1 E embodiment, 4 groups of Fig. 1 F embodiment, Fig. 1 G embodiment.
Fig. 2 xanthoxylum extract is to the anoxybiotic protective effect of neurocyte physical property
Fig. 2 A wherein: 5 groups of normal group, Fig. 2 B model group, 1 group of Fig. 2 C embodiment, 2 groups of Fig. 2 D embodiment, 3 groups of Fig. 2 E embodiment, 4 groups of Fig. 2 F embodiment, Fig. 2 G embodiment.
Fig. 3 Pericarpium Zanthoxyli polyphenol is to the protective effect of PC12 neurocyte 6-OHDA damage
Fig. 3 A wherein: 5 groups of normal group, Fig. 3 B model group, 1 group of Fig. 3 C embodiment, 2 groups of Fig. 3 D embodiment, 3 groups of Fig. 3 E embodiment, 4 groups of Fig. 3 F embodiment, Fig. 3 G embodiment.
Fig. 4 RT-PCR detects the influence that the xanthoxylum extract is expressed SR-B I mRNA
Wherein, a is GAPDH; B is SR-B I.
The specific embodiment
The preparation of embodiment 1 xanthoxylum extract
Get green pepper (Z.schinifolium Sieb.et Zucc.) peel 1000g, carry out reflux, extract, 3 times with 70% acetone 10L; Extracting solution is not gone up HPD500 type macroporous adsorptive resins through vacuum concentration after having the acetone flavor, wash impurity such as removing sugar earlier with water, use 60% ethanol elution then, it is that 1.10 back 95% ethanol with 5 times of amounts precipitate that eluent is concentrated into relative density, abandon precipitation, supernatant promptly gets brown ceramic powder shape xanthoxylum extract 62g through vacuum concentration, spray drying, the weight percentage of total polyphenols is 80% after testing, and wherein the weight percentage of ZP-CT-A is 15%.
The preparation of embodiment 2 xanthoxylum extracts
Get Fructus Zanthoxyli Plansipini (Z.armatum DC) peel 1000g, carry out reflux, extract, 3 times with 20L water; Extracting solution is directly gone up D101 type macroporous adsorptive resins, wash impurity such as removing sugar earlier with water, use 20% acetone eluting then, it is that 1.20 back 95% ethanol with 4 times of amounts precipitate that eluent is concentrated into relative density, abandon precipitation, supernatant promptly gets chocolate brown powder shape xanthoxylum extract 92g through vacuum concentration, spray drying, the weight percentage of total polyphenols is 50% after testing, and wherein the weight percentage of ZP-CT-A is 5%.
The preparation of embodiment 3 xanthoxylum extracts
Get Pericarpium Zanthoxyli (Z.bungeanum Maxim.) peel 1000g, carry out percolation with 70% ethanol 18L and extract; Extracting solution is not gone up HP20 type macroporous adsorptive resins through vacuum concentration after having the ethanol flavor, wash impurity such as removing sugar earlier with water, use 60% ethanol elution then, it is that 1.15 back 95% ethanol with 3 times of amounts precipitate that eluent is concentrated into relative density, abandon precipitation, supernatant promptly gets brown ceramic powder shape xanthoxylum extract 43g through vacuum concentration, spray drying, the weight percentage of total polyphenols is 74% after testing, and wherein the weight percentage of ZP-CT-A is 12%.
The preparation of embodiment 4 xanthoxylum extracts
Get Folium toonae sinensis Pericarpium Zanthoxyli (Z.ailantholides Sieb.et Zucc.) peel 1000g, carry out reflux, extract, 3 times with 40% ethanol 5L; Extracting solution is not gone up AB-8 type macroporous adsorptive resins through vacuum concentration after having the ethanol flavor, wash impurity such as removing sugar earlier with water, use 40% acetone eluting then, it is that 1.20 back 95% ethanol with 5 times of amounts precipitate that eluent is concentrated into relative density, abandon precipitation, supernatant promptly gets brown ceramic powder shape xanthoxylum extract 52g through vacuum concentration, spray drying, the weight percentage of total polyphenols is 65% after testing, and wherein the weight percentage of ZP-CT-A is 8%.
The preparation of embodiment 5 xanthoxylum extracts
Get Radix Zanthoxyli (Z.nitidum (Roxb.) DC.) peel 1000g, carry out reflux, extract, 3 times with 90% ethanol 10L; Extracting solution is not distinguished the flavor of to there being ethanol through vacuum concentration, add an amount of water-soluble even back of loosing and go up the polyamide adsorption resin column, wash with water earlier and remove non-phenol impurity, use 95% ethanol elution then, it is that 1.10 back 95% ethanol with 4 times of amounts precipitate that eluent is concentrated into relative density, abandons precipitation, supernatant is through vacuum concentration, spray drying, promptly get brown ceramic powder shape xanthoxylum extract 66g, the weight percentage of total polyphenols is 66% after testing, and wherein the weight percentage of ZP-CT-A is 8%.
The content assaying method of total polyphenols and ZP-CT-A:
The main reference of the assay of total polyphenols " Chinese pharmacopoeia (version in 2005) appendix X B carries out, and is specific as follows:
The preparation of P-Mo-Wo acid test solution: in the 1000ml flask, add 350ml water, 50g sodium tungstate (Na
2WO
42H
2O), 12.5g sodium molybdate (Na
2MoO
42H
2O), 25ml 50%H
3PO
4Aqueous solution, 50ml concentrated hydrochloric acid add several zeolites, reflux 10hr on electric furnace, and reuse 50ml washes out residue on the condensing tube, adds 75g Li
2SO
4, dripping several bromine waters again, opening boils 15min, and color finally becomes yellow, is chilled to the room temperature standardize solution to 500ml.
The preparation of reference substance solution: precision takes by weighing gallic acid reference substance 50mg and puts in the 100ml color, is dissolved in water and is diluted to scale, and precision is measured 5ml, puts in the brown measuring bottle of 50ml, and thin up shakes up to scale, promptly gets (containing gallic acid 0.05mg among every 1ml).
The preparation of standard curve: precision is measured reference substance solution 0.5ml, 1.0ml, 2.0ml, 3.0ml, 4.0ml, 5.0ml, put respectively in the brown measuring bottle of 25ml, each adds P-Mo-Wo acid test solution 1ml, add water 11.5ml, 11ml, 10ml, 9ml, 8ml, 7ml more respectively, be diluted to scale with 29% sodium carbonate liquor, shake up, placed 30 minutes, with the reagent corresponding is blank, with ultraviolet visible spectrophotometry, measure absorbance at 760nm wavelength place, be vertical coordinate with the absorbance, concentration is abscissa, the drawing standard curve.
The preparation of sample solution: precision takes by weighing Pericarpium Zanthoxyli polyphenol sample 50mg and puts in the 100ml brown bottle, is dissolved in water and is diluted to scale, and precision is measured 5ml, put in the brown measuring bottle of 50ml, thin up shakes up to scale, promptly gets (containing xanthoxylum extract 0.05mg among every 1ml).
Determining total phenol: precision is measured need testing solution 2ml, put in the brown measuring bottle of 25ml, method under the preparation of sighting target directrix curve, from " adding P-Mo-Wo acid test solution 1ml ", add water 10ml, measure absorbance in accordance with the law, from standard curve, read the amount (mg) of gallic acid in the need testing solution, calculate, promptly.
The content assaying method of ZP-CT-A (HPLC method):
Instrument: Waters 2690 HPLC, the Waters-996 diode is the row detector just; Waters symmetryC-18 chromatographic column, 4.6 * 250mm, 5 μ m;
Condition: detect wavelength: λ=280nm, flow velocity: 1.0mL/min, mobile phase: A:2% acetic acid, B: acetonitrile; Condition of gradient elution:
The preparation of embodiment 6 Pericarpium Zanthoxyli polyphenol extracts
Get pericarpium Zanthoxyli (Z.simulans Hance) peel 1000g, carry out reflux, extract, 3 times with 20% ethanol 5L; Extracting solution is not gone up XAD-4 type macroporous adsorptive resins through vacuum concentration after having the ethanol flavor, wash impurity such as removing sugar earlier with water, use 40% ethanol elution then, it is that 1.20 back ethanol with 5 times of amounts precipitate that eluent is concentrated into relative density, abandon precipitation, supernatant promptly gets brown ceramic powder shape Pericarpium Zanthoxyli polyphenol extract 49g through vacuum concentration, spray drying, total polyphenols content is 62% after testing, and wherein ZP-CT-A content is 6%.
The preparation of embodiment 7 Pericarpium Zanthoxyli polyphenol extracts
Get and rein in ﹠-Floor (Z.avicennae (Lam.) DC.) peel 1000g, carry out reflux, extract, 3 times with 20% acetone 5L; Extracting solution is not gone up HP-20 type macroporous adsorptive resins through vacuum concentration after having the ethanol flavor, wash impurity such as removing sugar earlier with water, use 20% ethanol elution then, it is that 1.20 back ethanol with 5 times of amounts precipitate that eluent is concentrated into relative density, abandon precipitation, supernatant promptly gets brown ceramic powder shape Pericarpium Zanthoxyli polyphenol extract 42g through vacuum concentration, spray drying, total polyphenols content is 51% after testing, and wherein ZP-CT-A content is 5%.
The isolation identification of embodiment 8 Pericarpium Zanthoxyli polyphenol compounds of the present invention
55g xanthoxylum extract (making) by embodiment 4 go up Diaion HP-20 chromatographic column (12 * 55cm, 1000g), water, 20%, 40%, 60%, 80%, 100% methanol-eluted fractions (each 1500mL) successively; 40% methanol-eluted fractions part (6.3g) is gone up Toyopearl HW-40C chromatographic column, and (7 * 12cm 250g), uses 10%, 20%, 30%, 40%, 50%, 60%, 70% methanol aqueous solution eluting (each 500mL) to obtain 7 sections (FrC1-7) successively; FrC2 (0.3g) is gone up MCI gel CHP-20P chromatographic column (4 * 9cm, 25g), water, 10%, 20%, 30%, 100% methanol-eluted fractions (each 50mL) successively, obtain 3-oxygen-caffeoylquinic acids (2) (26mg) and 4-oxygen-caffeoylquinic acids (3) (7.6mg); With FrC6 (120mg) go up MCI gel CHP-20P chromatographic column (1 * 20cm, 10g), water, 5%, 10%, 100% methanol-eluted fractions (each 30mL) obtain procyanidin B1 (4) (16mg) successively; 60% methanol-eluted fractions part (11g) goes up Toyopearl HW-40C chromatographic column (7 * 30cm on the Diaion HP-20 chromatographic column, 500g), successively use 70% ethanol water, 70% aqueous acetone solution eluting (each 1000mL), wherein 70% ethanol elution part (0.3g) goes up YMC gel ODS AQ 120-S50 chromatographic column (4 * 9cm, 25g), water successively, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 100% methanol-eluted fractions (each 50mL), from 30% methanol-eluted fractions thing, obtain hyperin (5) (31mg), from 70% aqueous acetone solution eluting part, directly obtain ZP-CT-A (1) (6.4g).
3-oxygen-caffeoylquinic acids (2), C
16H
18O
9Mp 202-204 ℃; UV λ
Max=325,299,245nm; APCI MS, m/z 355[M+1]
+ 1H-NMR (600MHz, CD
3OD): δ: 1.95 (1H, dd, J=9,14Hz, H-6ax), 2.13 (2H, m, H-2eq and 6eq), 2.20 (1H, dd, J=4,15Hz, H-2ax), 3.63 (1H, dd, J=3,9Hz, H-4), 4.14 (1H, ddd, J=3,9,9Hz, H-5), 5.34 (1H, ddd, J=3,3,4Hz, H-3), 6.30 (1H, d, J=16Hz, H-8 '), 6.76 (1H, d, J=8Hz, H-5 '), 6.93 (1H, dd, J=2,8Hz, H-6 '), 7.04 (1H, d, J=2Hz, H-2 '), 7.58 (1H, d, J=16Hz, H-7 ');
13C-NMR (150MHz, CD
3OD): δ: 36.7 (C-2), 41.5 (C-6), 68.3 (C-5), 73.0 (C-3), 74.8 (C-4), 75.4 (C-1), (115.1 C-2 '), 115.8 (C-8 '), 116.4 (C-5 '), (122.9 C-6 '), (127.9 C-1 '), 146.79 (C-3 '), 146.80 (C-7 '), (149.4 C-4 '), (169.0 C-9 '), 178.3 (C-7). the data consistent of above data and bibliographical information (Nobuji Nakatani, Shin-ichiKayano et al.Identification, quantitative determination, and antioxidativeactivities of chlorogenic acid isomers in Prune (Prunus domestica L.) .J.Agric.Food Chem.2000; 48:5512-5516).
4-oxygen-caffeoylquinic acids (3), C
16H
18O
9Mp 203-205 ℃; UV λ
Max=326,300,245nm; APCI MS, m/z 355[M+1]
+ 1H-NMR (600MHz, CD
3OD): δ: 2.00 (1H, dd, J=11,13Hz, H-6ax), 2.06 (1H, ddd, J=3,4,14Hz, H-2eq), 2.17 (1H, dd, J=4,14Hz, H-2ax), 2.20 (1H, ddd, J=3,5,13Hz, H-6eq), 4.27 (1H, ddd, J=4,9,11Hz, H-5), 4.28 (1H, ddd, J=3,3,4Hz, H-3), 4.79 (1H, dd, J=3,9Hz, H-4), 6.37 (1H, d, J=16Hz, H-8 '), 6.77 (1H, d, J=8Hz, H-5 '), 6.96 (1H, dd, J=2,8Hz, H-6 '), 7.06 (1H, d, J=2Hz, H-2 '), 7.65 (1H, d, J=16Hz, H-7 ');
13C-NMR (150MHz, CD
3OD): δ: 38.4 (C-2), 42.7 (C-6), 65.5 (C-5), 69.6 (C-3), 76.6 (C-1), 79.3 (C-4), (115.1 C-2 '), 115.4 (C-8 '), 116.5 (C-5 '), (123.0 C-6 '), (127.8 C-1 '), 146.8 (C-3 '), 147.1 (C-7 '), (149.6 C-4 '), (169.0 C-9 '), 177.3 (C-7). the data consistent of above data and bibliographical information (Nobuji Nakatani, Shin-ichi Kayano et al.Identification, quantitative determination, and antioxidative activities of chlorogenic acidisomers in Prune (Prunus domestica L.) .J.Agric.Food Chem.2000; 48:5512-5516).
Procyanidin B1 (4), C
30H
26O
12The unformed powder of white, anisaldehyde-concentrated sulphuric acid shows orange colour; [α]
D 20:+14.0 ° (c 0.55, in MeOH); APCI MS (positive) m/z 579[M+1]
+(negative) m/z 577[M-1]
- 1H-NMR (CD
3OD): δ: 2.82 (d, J=15.0Hz), 2.92 (dd, J=15.0,4.0Hz), 3.88 (s), 4.30 (s), 4.62 (s) 4.80 (s), 5.79 (d, J=2.0Hz), 5.83 (d, J=2.0Hz), 5.89 (d, J=2.5Hz), 5.93 (d, J=2.5Hz), 6.61 (d, J=7.0Hz), 6.67 (dd, J=7.0Hz), 6.72 (d, J=2.0Hz), 6.79 (d, J=7.5Hz), 6.85 (dd, J=7.5,2.0Hz), 7.08 (d, J=2.0Hz).
13C-NMR (CD
3OD): δ: 29.6,37.1,66.9,73.4,77.0,79.7,96.1,96.5,97.2,100.5,101.3,107.4,115.6-115.9,119.3,119.4,132.0,132.6,145.5-145.8,156.3-158.0. the data consistent of above data and bibliographical information (R.S.Thompson, D.Jacques et al., Plant proanthocyanidins.Part I.Introduction; The isolation, structure, and distribution in nature of plant procyanidins.J.Chem.Soc., PerkinTrans.1,1972,1387-1399; Zhang Chaofeng, during Sun Qi etc., tannin constituents and anti-HIV-1 intergrase activity research thereof in the Radix Linderae stem. Chinese Pharmaceutical Journal .2003; 38 (12): 911-914).
Hyperin (5), C
21H
20O
12The crystallization of yellow particle shape, magnesium powder-hydrochloric acid reaction show red, and it is green that ferric chloride reaction shows, the alphanaphthol reaction positive; Mp 232-233 ℃; [α]
D 20:-83 ° (c 0.2, inpyridine); UV λ
Max=363nm;
1H-NMR (DMSO-d
6): δ: 12.63 (1H, s, OH-5), 10.86 (1H, s, OH-7), 9.64 (1H, s, OH-4 '), 9.35 (1H, s, OH-3 '), 7.54 (1H, d, J=1.2Hz, H-2 '), 7.46 (1H, dd, J=1.2,8.4Hz, H-6 '), 6.84 (1H, d, J=8.4Hz, H-5 '), 6.40 (1H, d, J=1Hz, H-8), 6.19 (1H, d, J=1Hz, H-6), 5.42 (1H, d, J=1.2Hz), 3.30-4.28 (5H, the sugar ring is gone up proton).The data consistent of above data and bibliographical information (Yu Dequan, Yang Junshan. analytical chemistry handbook spectral analysis of the nuclear magnetic resonance [M]. the 2nd edition. Beijing: Chemical Industry Press, 1999:825; Yao new life. Natural Medicine Chemistry [M]. the 2nd edition. Beijing: the People's Health Publisher, 1998:102).
The structural identification of ZP-CT-A:
ZP-CT-A (1), light brown powder,
13C-NMR[150MHz, Me
2CO-d
6-D
2O (7: 3); 40 ℃]: δ: 26-39 (C-4, C-3), 68-81 (C-3, C-2), 95-112 (C-4a, C-6, C-8), 113-122 (C-2 ', C-5 ', C-6 '), 128-134 (C-1 '), 143-147 (C-3 ', C-4 '), 151-159 (C-5, C-7, C-8a); Results of elemental analyses: C, 54.37%; H, 5.33%, near C
15H
12O
62.5H
2The value of calculation of O: C, 54.06%; H, 5.14%.
ZP-CT-A's
13The C-NMR data are the similar (Czochanska of unitary poly procyanidin to bibliographical information with catechin and/or epicatechin, Z., Foo, L.Y.et al., Polymericproanthocyanidins:stereochemistry, structural units, and molecular weight.J.Chem.Soc., Perkin Trans.1980; 1:2278-2286).
Efficient volume exclusion chromatography (HP-SEC) is measured the ZP-CT-A molecular weight: adopt TSK gelSuperAW3000 chromatographic column (6.0 * 150mm; Tosoh), mobile phase is N, and dinethylformamide (DMF) wherein contains 0.5% 3M ammonium formate (HCOONH
4), 40 ℃ of column temperatures, flow velocity 0.5mL/min detects wavelength 280nm, is the contrast that molecular weight calculates with epicatechin, procyanidin B1, Cortex cinnamomi japonici (Ramulus Cinnamomi) tannin A2, is calculated the weight average molecular weight (M of ZP-CT-A by size exclusion chromatograph figure
w) and number-average molecular weight (M
n) (Bae, Y.S., Foo, L.Y.et al., GPC of natural procyanidinoligomers and polymers.Holzforschung, 1994; 48:4-6), the result is respectively M
w=6.4 * 10
3, M
n=5.0 * 10
3, hence one can see that, and ZP-CT-A is the poly procyanidin of 17-mer.
The ZP-CT-A polymerized unit can further be determined by the thiolysis product of measuring ZP-CT-A:
The thiolysis of ZP-CT-A (employing contains the acetic acid of 1-lauryl mercaptan): get 10mgZP-CT-A, 50 μ L 1-lauryl mercaptans, 125 μ L acetic acid and 1.25mL ethanol respectively in the 5mL glass ampoule, mixing, seal with alcohol blast burner, place 100 ℃ of water-baths to react 6hr, product is analyzed with HPLC, and concrete grammar is as follows:
Instrument: Waters 2690 HPLC, the Waters-996 diode is the row detector just; Waters symmetryC-18 chromatographic column, 4.6 * 250mm, 5 μ m;
Condition: column temperature: 40 ℃, detect wavelength: λ=280nm, flow velocity: 1.0mL/min, mobile phase: A:0.01M H
3PO
4-0.01M KH
2PO
4-CH
3CN (45.25: 45.25: 9.5), B:0.01MH
3PO
4-0.01M KH
2PO
4-CH
3CN (15: 15: 70); Condition of gradient elution: 0-20min, 100% A, 20-50min, 100%B.
The HPLC analysis result shows, contains catechin, epicatechin, catechin-4 α-dodecyl thioether (6), epicatechin-4 β-dodecyl thioether (7) in the product, wherein the 6, the 7th, and main derivative products.By the molar ratio that can determine 6,7 with respect to the peak area of catechin is 1: 5.1.
(MD) chromatographic column is with [CHCl for Sephadex, Columbia for Sephadex LH-20 on the reactant mixture of 100mg ZP-CT-A after said method is degraded
3-EtOH (1: 1) → EtOH] gradient elution, further carry out purification on the HPLC partly preparing again with YMC A324 chromatographic column, can obtain catechin-4 α-dodecyl thioether (6) (2.2mg), epicatechin-4 β-dodecyl thioether (7) (16.3mg).
Catechin-4 α-dodecyl thioether (6): the unformed powder of light brown, ESI-MS:m/z:491 (M+H)
+.HR-ESI-MS m/z:491.2458 (C
27H
38O
6The S+H value of calculation is 491.2467).
1H-NMR(600MHz，DMSO-d
6)：δ：0.83(3H，t，J＝7Hz，H-12″)，1.25(16H，br，H-4″-H-11″)，1.36(2H，m，H-3″)，1.60(2H，m，H-2″)，2.73，2.84(1H，each，dt，J＝12，7Hz，H-1″)，4.04(1H，dd，J＝4，10Hz，H-3)，4.29(1H，d，J＝4Hz，H-4)，4.86(1H，d，J＝10Hz，H-2)，5.78，6.01(1H?each，d，J＝2Hz，H-6，H-8)，6.75(2H，m，H-5′，H-6′)，6.88(1H，s，H-2′).
Epicatechin-4 β-dodecyl thioether (7), the unformed powder of light brown, ESI-MS:m/z:491 (M+H)
+.HR-ESI-MS m/z:491.2446 (C
27H
38O
6The S+H value of calculation is 491.2467).
1H-NMR(600MHz，DMSO-d
6)：δ：0.81(3H，t，J＝7Hz，H-12″)，1.22(16H，br，H-4″-H-11″)，1.3-1.4(2H，m，H-3″)，1.62(2H，m，H-2″)，2.6-2.7(2H，m，H-1″)，3.93(2H，s，H-3，H-4)，5.22(1H，s，H-2)，5.86，5.95(1H?each，d，J＝2.5Hz，H-6，H-8)，6.7-6.4(2H，m，H-5′，H-6′)，7.01(1H，br?s，H-2′).
13C?NMR：δ：13.8(C-10″)，?23.0(C-9″)，28.5-32.4(C-1″-C-8″)，42.9(C-4)，71.2(C-3)，74.9(C-2)，95.1，96.3(C-6，C-8)，99.7(C-10)，114.8，115.1(C-2′，C-5′)，118.8(C-6′)，131.5(C-1′)，144.9，145.0(C-3′，C-4′)，156.5，158.0，158.4(C-5，C-7，C-9).
The above analysis can determine that ZP-CT-A (1) is the structure in the diagram, and its polymerized unit is mainly epicatechin, and the unit, end is catechin and epicatechin (molar ratio is 1: 0.8).
Embodiment 9 preparation tablets
With xanthoxylum extract 100g of the present invention (embodiment 1 method makes), starch 80g, dextrin 5g mix homogeneously adds 10% starch slurry system soft material, granulates with 14 order nylon screens, 60-70 ℃ of aeration-drying, 16 mesh sieve granulate add magnesium stearate 1.5g, carboxymethyl starch sodium 5g mixing, be pressed into 1000, coating promptly.Every contains xanthoxylum extract 100mg.Become human oral every day 2 times, each 2.
The preparation of embodiment 10 capsules
Get xanthoxylum extract 100g of the present invention (embodiment 2 methods make), add starch 78g, magnesium stearate 2g mixing directly is filled to 1000 with Autocapsulefillingmachine, and polishing promptly.Become human oral every day 2 times, each 2.
The preparation of embodiment 11 drop pill
Get xanthoxylum extract 10g of the present invention (embodiment 3 methods make), drop in the polyethylene glycol 6000 of 32g heating and melting, be stirred to dissolving, be transferred in the reservoir, airtight and insulation is regulated drop pill machine drop quantitative valve at 80-90 ℃, splash into from top to bottom in 10-15 ℃ the liquid paraffin, make 1000 altogether, the ball sound of rain pattering dry doubling erasing liquor paraffin body with forming is drying to obtain.Every contains xanthoxylum extract 10mg.Become human oral every day 3 times, each 10-15 grain.
The preparation of embodiment 12 oral liquids
Get xanthoxylum extract 20g of the present invention (embodiment 4 methods make), mix with Mel 300g, sucrose 50g, sodium benzoate 2g and distilled water 300ml, be heated to 85-90 ℃, stirring makes dissolving, and insulation 30min filters, the filtrate thin up is to 1000ml, stir evenly, embedding (every 10ml), sterilization is promptly.Become human oral every day 2 times, each 1.
The preparation of embodiment 13 granules
It is an amount of to get xanthoxylum extract 10g of the present invention (embodiment 5 methods make), dextrin 20g, sucrose 100g and ethanol, and mixing is crossed 10 mesh sieves and made granule, in 60-70 ℃ of drying, granulate, packing promptly, the heavy 5g of every bag becomes human oral every day 1 time, each 1 wraps.
The preparation of embodiment 14 injection
Get xanthoxylum extract 100g of the present invention (embodiment 3 methods make), add water for injection and make dissolving in right amount, 0.02% the active carbon that adds amount of preparation stirs 5-10min, filter, filtrate is diluted to about 10 liters, adds sodium chloride adjusting osmotic pressure and oozes to waiting, and regulates pH7.5-8.0, ultrafiltration, embedding become 1000 (10mL/ props up).100 ℃ of 30min sterilizations promptly.Adult's vein or administered intramuscular, every day 2 times, each 1.
The preparation of embodiment 15 injectable powder
Get xanthoxylum extract 100g of the present invention (embodiment 1 method makes), add injection water and dilute sodium hydroxide and make dissolving in right amount, 0.02% the active carbon that adds amount of preparation stirs 5-10min, filter, filtrate is diluted to 1 liter, regulates pH6.5-7.8, ultrafiltration, spray drying, dry powder is promptly aseptic subpackaged.Every 100mg faces with before adding the injection water and makes dissolving in right amount, with the slowly intravenous drip of sodium chloride transfusion 250-500mL dilution back.Adult every day 2 times, each 1.
Below to test the formal proof beneficial effect of the present invention of example.
Test example 1 xanthoxylum extract is to the inhibitory action of neutral endopeptidase (NEP)
NEP is the important target spot of antihypertensive drug of having generally acknowledged at present, suppresses its activity, can effectively reduce blood pressure.Experiment showed, that below the xanthoxylum extract has the activity that suppresses NEP, thereby can bring high blood pressure down.Contain an amount of neutral endopeptidase (from the rabbit nephridial tissue, extracting), pH7.4 Tris-HCl buffer, ACE inhibitor Captopril (SIGMA), APN inhibitor B estatin (SIGMA) and sample in the 200 μ l reaction systems, set up blank (not containing enzyme and sample) and negative control (not containing sample) simultaneously, 37 ℃ of reaction 30min, add neutral endopeptidase substrate DAGNPG (SIGMA) solution, 37 ℃ are reacted 60min again, measure fluorescence intensity F, excitation wavelength 355nm, emission wavelength 560nm.Calculate suppression ratio, suppression ratio=[1-(F sample-F blank)/(F feminine gender-F blank)] * 100% according to fluorescence intensity F value.The result shows that the xanthoxylum extract all has inhibitory action to NEP. the xanthoxylum extract of embodiment 1-5 preparation is to the IC of NEP
50Be respectively: 90 μ g/ml, 112 μ g/ml, 96 μ g/ml, 107 μ g/ml, 101 μ g/ml.
Test example 2 xanthoxylum extracts are to the anoxybiotic protective effect of vascular endothelial cell physical property.
Cultivate human vascular endothelial strain Eahy926 cell, in inoculating cell to 96 orifice plate, overnight incubation is inhaled and is abandoned culture medium, and the normal control group adds Glucose-Hanks, 37 ℃, CO
2The incubator incubated overnight; The anoxia model group adds Hanks, and the drug treating group adds Hanks and Pericarpium Zanthoxyli polyphenol (50 μ g/ml), and the two all puts into closed enclosure, logical 95%N
2, 5%CO
2, after lasting 5 minutes (5L/min), 37 ℃ of overnight incubation.Measure cell density with mtt assay then, calculate the suppression ratio of medicine, suppression ratio=(medicine group-model group)/(normal group-model group) damage.Result (table 1, Fig. 1) shows, the xanthoxylum extract of embodiment 1-5 preparation all has protective effect, each medicine group to compare with model group to damage due to the Eahy926 vascular endothelial cell physical property anoxia significant difference (P＜0.05) is all arranged.
Table 1 xanthoxylum extract is to the anoxybiotic protective effect of vascular endothelial cell physical property.
| Group | Concentration (μ g/ml) | Cell density (OD value) | Damage suppression ratio (%) |
| Normal group | ? | 0.422±0.011 | / |
| Model group | ? | 0.179±0.039 ** | / |
| Embodiment 1 Pericarpium Zanthoxyli polyphenol | 50 | 0.337±0.008 △ | 65.02±6.03 |
| Embodiment 2 Pericarpium Zanthoxyli polyphenol | 50 | 0.275±0.01 △ | 39.55±4.44 |
| Embodiment 3 Pericarpium Zanthoxyli polyphenol | 50 | 0.318±0.004 △ | 57.20±5.66 |
| Embodiment 4 Pericarpium Zanthoxyli polyphenol | 50 | 0.282±0.022 △ | 42.66±4.75 |
| Embodiment 5 Pericarpium Zanthoxyli polyphenol | 50 | 0.300±0.032 △ | 49.91±4.89 |
Annotate:
*Expression is compared P＜0.01 with normal group;
△Expression is compared P＜0.05 with model group.
Test example 3 xanthoxylum extracts are to the anoxybiotic protective effect of neurocyte physical property.
Cultivate Mus pheochromocytoma neurocyte strain PC12 cell, in inoculating cell to 96 orifice plate, overnight incubation is inhaled and is abandoned culture medium, and the normal control group adds Glucose-Hanks, 37 ℃, CO
2The incubator incubated overnight; The anoxia model group adds Hanks, and the drug treating group adds Hanks and Pericarpium Zanthoxyli polyphenol (50 μ g/ml), and the two all puts into closed enclosure, logical 95%N
2, 5%CO
2, after lasting 5 minutes (5L/min), 37 ℃ of overnight incubation.Measure cell density with srb assay then, calculate the suppression ratio of medicine, suppression ratio=(medicine group-model group)/(normal group-model group) damage.Result (table 2 and Fig. 2) shows, the xanthoxylum extract of embodiment 1-5 preparation all has protective effect, each medicine group to compare with model group to damage due to the PC12 neurocyte physical property anoxia significant difference (P＜0.05) is all arranged
Table 2 xanthoxylum extract is to the anoxybiotic protective effect of neurocyte physical property
| Group | Concentration (μ g/ml) | Cell density (OD value) | Damage suppression ratio (%) |
| Normal group | ? | 0.251±0.006 | /15 |
| Model group | ? | 0.124±0.002 * | / |
| Embodiment 1 Pericarpium Zanthoxyli polyphenol | 50 | 0.212±0.01 △ | 69.44±6.03 |
| Embodiment 2 Pericarpium Zanthoxyli polyphenol | 50 | 0.166±0.008 △ | 33.41±3.23 |
| Embodiment 3 Pericarpium Zanthoxyli polyphenol | 50 | 0.195±0.004 △ | 56.03±6.66 |
| Embodiment 4 Pericarpium Zanthoxyli polyphenol | 50 | 0.179±0.022 △ | 43.91±3.75 |
| Embodiment 5 Pericarpium Zanthoxyli polyphenol | 50 | 0.184±0.018 △ | 47.29±3.89 |
Annotate:
*Expression is compared P＜0.05 with normal group,
△Expression is compared P＜0.05 with model group.
Test example 4 xanthoxylum extracts are to the protective effect of neurocyte 6-OHDA damage
Cultivate Mus pheochromocytoma neurocyte strain PC12 cell, be seeded to 96 orifice plates, overnight incubation is inhaled and is abandoned culture medium, carries out the 6-OHDA injury experiment.To add normal cultivation basis set is normal control, to contain 60uM 6-OHDA is model group, each hole of medicine group is 60uM 6-OHDA+ xanthoxylum extract (50 μ g/ml), 37 ℃ of overnight incubation, measure cell density with srb assay, calculate the suppression ratio of medicine pair cell damage, suppression ratio=(medicine group-model group)/(normal group-model group).Result such as table 3 and shown in Figure 3, as seen, the xanthoxylum extract of embodiment 1-5 preparation all has significant protective effect, each medicine group to compare with model group to damage due to the PC12 neurocyte 6-OHDA significant difference (P＜0.05) is all arranged.
Table 3 Pericarpium Zanthoxyli polyphenol is to the protective effect of PC12 neurocyte 6-OHDA damage
| Group | Concentration (μ g/ml) | Cell density (OD value) | Damage suppression ratio (%) |
| Normal group | ? | 0.274±0.009 | / |
| Model group | ? | 0.172±0.008 ** | / |
| Embodiment 1 Pericarpium Zanthoxyli polyphenol | 50 | 0.239±0.006 △ | 66.47±4.63 |
| Embodiment 2 Pericarpium Zanthoxyli polyphenol | 50 | 0.210±0.007 △ | 37.32±3.40 |
| Embodiment 3 Pericarpium Zanthoxyli polyphenol | 50 | 0.231±0.002 △ | 57.37±6.80 |
| Embodiment 4 Pericarpium Zanthoxyli polyphenol | 50 | 0.219±0.021 △ | 46.09±4.45 |
| Embodiment 5 Pericarpium Zanthoxyli polyphenol | 50 | 0.222±0.01 △ | 49.36±3.49 |
Annotate:
*Expression is compared P＜0.01 with normal group;
△Expression is compared P＜0.05 with model group.
Test example 5 xanthoxylum extracts increase hepatocyte HDL receptor SR-B I expresses
Present known SR-B I is treatment hyperlipidemia and the pharmaceutically-active important target spot of atherosclerosis, promotes the high density lipoprotein of its expression in can elevating blood, blood viscosity lowering.Experimental results show that below the Pericarpium Zanthoxyli polyphenol can promote SR-B I to express, thus can hyperlipidemia and treatment atherosclerosis.Cultivate the HepG2 hepatoma carcinoma cell, add the Fructus Zanthoxyli Plansipini polyphenol extract of 100 μ g/mL embodiment, 2 preparations, effect 24hr uses the expression that sxemiquantitative RT-PCR detects HDL receptor.KODAK 1D software analysis is used in agarose gel electrophoresis (as Fig. 4) back of taking pictures, optical density (OD) value according to SR-B I purpose band and internal reference GAPDH, represent the SR-BI expression with the relative ratio of SR-B I/GAPDH, the result, blank group and administration group ratio are respectively: 0.47,0.64, illustrate that this xanthoxylum extract can strengthen the expression of HDL receptor SR-B I mRNA.
The blood coagulation resisting function of test example 6 xanthoxylum extracts
Kunming mouse 18-20g is divided into each group of normal group and embodiment 1-5 extract (60mg/kg) at random, and xanthoxylum extract gastric infusion, administration time are 21 days, detect the clotting time of animal respectively after administration in 10,14,21 days with capillary tube method.Experimental data is checked with T and is carried out statistical analysis.The result is as shown in table 3, and administration is after 14 days, 21 days as can be seen from Table 3, and each embodiment extract group clotting time all has the prolongation effect than normal group, and has compared significant difference with normal group (P＜0.05), illustrates that each extract all has blood coagulation resisting function.
Each embodiment xanthoxylum extract of table 4 is to the influence of normal clotting time of mice
Annotate: compare with normal group,
*Expression P＜0.05,
*Expression P＜0.01.
Above-mentioned pharmacodynamics test explanation, xanthoxylum extract drug effect of the present invention is that index limits with total polyphenols and procyanidin polymer ZP-CT-A, be 50～80% at total polyphenols content, the weight percentage of procyanidin polymer ZP-CT-A in extract be 5%～15% o'clock, has definite drug effect, especially the total polyphenols percentage by weight is 65%～80%, the weight percentage of procyanidin polymer ZP-CT-A in extract is 8%～15% o'clock, stable drug effect is arranged, and drug effect has significant significant difference.
Above-mentioned results of pharmacodynamic test shows; xanthoxylum extract energy neuroprotective cell of the present invention avoids anoxia-induced apoptosis and 6 hydroxy dopamine cause damage; the protection vascular endothelial cell is avoided anoxia-induced apoptosis; suppress the neutral endopeptidase activity, raise HDL receptor SR-B I expression; and have blood coagulation resisting function, can be used as the preparation nerve cell-protective agents, resist myocardial ischemia or a kind of new selection of cerebral ischemia class medicine, anticoagulant, neutral endopeptidase inhibitor and lipidemia class medicine.
Claims (7)
1. the xanthoxylum peel extract is avoided anoxia-induced apoptosis and 6 hydroxy dopamine cause damage at preparation neuroprotective cell, and the protection vascular endothelial cell is avoided anoxia-induced apoptosis, raises HDL receptor SR-B
Application in I expression, anticoagulant, the neutral endopeptidase inhibitor medicaments;
The preparation method of described xanthoxylum peel extract comprises the following steps:
A, extraction: with peel water or the aquiferous ethanol or the aqueous acetone extraction of xanthoxylum, raw material and extracting solution w/v are 1: 5～20;
B, concentrated: extracting solution is not distinguished the flavor of to having alcohol flavor or acetone through vacuum concentration;
C, absorption with macroporous adsorbent resin, eluting: above-mentioned concentrated solution is added water-soluble diffusing back go up macroporous adsorptive resins or polyamide, first water eluting is used 20%-95% ethanol or acetone eluting then;
D, precipitate with ethanol: the ethanol with 3～5 times of amounts after eluent concentrates precipitates, and abandons precipitation;
E, concentrated, dry: supernatant concentration, drying promptly get the xanthoxylum peel extract;
Described xanthoxylum peel extract is from Pericarpium Zanthoxyli (Zanthoxylum bungeanum Maxim.), green pepper (Z.schinifolium Sieb.et Zucc.), Fructus Zanthoxyli Plansipini (Z.armatum DC), pericarpium Zanthoxyli (Z.simulans Hance), reins in to extract the peel of ﹠-Floor (Z.avicennae (Lam.) DC.), Folium toonae sinensis Pericarpium Zanthoxyli (Z.ailantholides Sieb.et Zucc), a green pepper (Z.molle Rehd) or Radix Zanthoxyli (Z.nitidum (Roxb.) DC.) and obtain.
2. purposes according to claim 1 is characterized in that: containing the total polyphenols percentage by weight in the described peel extract is 50%～80%.
3. purposes according to claim 2 is characterized in that: the total polyphenols percentage by weight is 65%～80%.
4. according to claim 2 or 3 described purposes, it is characterized in that: contain procyanidin polymer ZP-CT-A in the total polyphenols, wherein, the weight percentage of procyanidin polymer ZP-CT-A in peel extract is 5%～15%.
5. purposes according to claim 4 is characterized in that: the weight percentage of described procyanidin polymer ZP-CT-A in peel extract is 8%～15%.
6. purposes according to claim 1 is characterized in that: described medicine is that the xanthoxylum peel extract with effective dose is an active component, adds the medicament that acceptable accessories or complementary composition are prepared from.
7. purposes according to claim 6 is characterized in that wherein said medicament is oral formulations, injection preparation.
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| CN102423358A (en) * | 2011-11-18 | 2012-04-25 | 邱宏亮 | Method for extracting total alkaloids from Zanthoxylum simulans Hance rhizome and medicinal composition for oral cavity |
| CN102836260B (en) * | 2012-09-07 | 2014-06-11 | 四川大学 | Method for extracting plant polyphenol with antioxidant activity from wild pepper leaves |
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