CN101274011A - Preparation of walnut shell extract and applications thereof in preparations of cardiotonic medicine and cardio-cerebrovascular medicine - Google Patents
Preparation of walnut shell extract and applications thereof in preparations of cardiotonic medicine and cardio-cerebrovascular medicine Download PDFInfo
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Abstract
The invention discloses a preparation method for extracting ethanol from an extract of a natural walnut shell, a pharmaceutical composition using the extract as the active component and research results of relevant pharmacology. A walnut shell is ground and boiled for extracting the ethanol and then is concentrated and dried for obtaining the brown red extract. The extracting method is characterized by short process, relatively high extracting rate and is easy for industrialized production. The discovery of the medical value of the walnut shell and the development of the product can prevent great waste of natural pharmaceutical resources. The research of the pharmacological activity shows that the walnut shell extract has a certain activity for strengthening heart and improving myocardial blood supply and is possible to be developed into a new pharmaceutical of natural traditional Chinese medicine for curing cardio-cerebrovascular disease, having a plurality of pharmacological activities, low toxicity and high efficiency. Various oral preparations of different pharmaceutical compositions can be prepared by mixing the extract of effective curing amount with pharmaceutical adjuvant.
Description
One, technical field:
The present invention belongs to field of medicaments, particularly natural pharmacology extraction of active ingredients and in the purposes of heart and cardiovascular disease prevention pharmaceutical preparation in the natural medicine field.The present invention relates to a kind of natural nucleus peach shell extraction of active ingredients preparation method, more specifically saying so obtains the active component group through water or certain density ethanol extraction from Semen Juglandis leftover bits and pieces walnut shell, this active component group is rich in Semen Juglandis quinone, its glycosides, tannin, polysaccharide and vegetable protein constituents.The invention discloses it and extract preparation method and be the pharmaceutical composition of active component with this extract, and said composition the preparation heart tonifying and improve myocardial ischemia and cardiovascular and cerebrovascular disease control pharmaceutical preparation in application.This extraction process is the technology of preparing of natural product in the Medicines and Health Product field.But it has disclosed the Technology that preparation is extracted in a kind of industrialization.
Two, background technology:
Walnut shell is the duricrust of juglandaceae plant Semen Juglandis Juglans reglia L. mature fruit.Semen Juglandis has another name called Semen Juglandis, Qiang peach, is used as medicine with Semen Juglandis usually, has the kidney invigorating and essence nourishing, effects such as warming the lung Dingchuan.Semen Juglandis has the cultivation history in more than 2000 year in China, and the country that produces Semen Juglandis in the world is a lot, except that China, also has states such as U.S., method, meaning, India, Turkey; Wherein output, area maximum is the China and the U.S., adds up to annual production to surpass 80% of Gross World Product.Walnut all has cultivation to distribute in China big extremely total points provinces and regions, area has more than 1,000 ten thousand mu, and natural resources is very abundant.After the eighties in 19th century, China's Semen Juglandis output is the trend that grows steadily.Semen Juglandis annual production in 2002 has reached 300,000 tons.Along with the popularizing planting of high-quality seeds, Semen Juglandis output is also increasing year by year.
Semen Juglandis is nutritious, and protein and unsaturated fatty acid content height are particularly suitable for the absorption of human body utilization, have higher nourishing healthy and medical value.Semen Juglandis can be used as health food and is processed into various cuisines.Along with the development of Semen Juglandis processing industry in recent years, a large amount of walnut shell offals are collected utilization.Semen Juglandis processing kernel percent calculates by 45～50%, and the Semen Juglandis hatching rate reaches 50～55%, and conversion is got up can have 15～200,000 tons walnut shell resource to utilize present every year.Walnut shell is main directly as the raw material of producing active carbon now, and make the environmental protection filtrate and carry out sewage disposal, and application, grinding-material manufacturing etc. in the oil field, walnut shell comes research and utilization also seldom as medicine at present.Modern study shows that walnut shell has antioxidant activity, can remove the OH-free radical effectively, and to O
2-no scavenging action can compare favourably with the tea polyphenols with concentration.But simple antioxidant activity, very difficult definite its used in which kind of disease can have better prevention to tire.The research that the pharmacologically active of walnut shell, function cure mainly is at home and abroad carried out also seldom, influences effective development and utilization that this enriches natural resources, causes the huge waste of this drug resource for a long time.
We know that cardiovascular and cerebrovascular disease is still the formidable enemy of human health, are old common disease and frequently-occurring disease, and M ﹠ M still occupies first of all kinds of major diseases.Treat natural cardiac tonic extensive use of heart failure clinically based on digitaloid drugs (as digoxin), but this class drug toxicity is very big, therapeutic domain is narrow, effective dose and toxic dose are approaching, be easy to cause poisoning, cause a hidden trouble for the safety of medication, patient cannot use for a long time in a large number.Therefore seek the effective cardiac drug of natural low toxicity, to heart patient and the medication of patients with heart failure long-term safety, the puzzlement that throws off one's illness is significant.Moreover deficiency myocardial blood supply is to cause heart and injury and chronic functional to decay, move towards depleted major reason gradually, and improving blood supply of cardiac muscle is the critical treatment measure that reduces and prevent relevant disease to take place, develop.Equally, the medicine of blood fat reducing can effectively be prevented and treated arteriosclerotic generation, and to the control cardiovascular and cerebrovascular disease, defying age is significant
The present invention is a raw material with Semen Juglandis leftover bits and pieces walnut shell, through selecting, pulverize, sieve, alcohol reflux, reclaim solvent, drying obtains the effective ingredient porphyrize, operations such as preparation, preparation and getting.Medicine contains Semen Juglandis quinone and compositions such as glycosides, tannin thereof.It is simple to have technology, composition and efficacy stability, the feature that easy realization of industrialization is produced.Its cardiotonic and improve the myocardial ischemia pharmacologically active and be first the new pharmacologically active of finding; Also have certain blood fat reducing and blood pressure regulation effect in addition, can be used for preventing and treating arteriosclerosis.Thereby can be used for the application of cardiovascular disease prevention medicine.
Pharmacology activity research shows, the total extract of alcohol extraction and water extraction has cardiac activity and improves the effect of blood supply of cardiac muscle, and to the liver spleen kidney not damaged of body, do not damage immunologic function, might become fully and have good heart disease and the new traditional Chinese medicinal materials assortment of cardiovascular disease prevention effect.Heart is active simultaneously body is free from side effects substantially in performance, is a kind of natural plant composition medicine with control heart failure and cardiovascular and cerebrovascular disease effect, and this also is a pharmacologically active of finding that first walnut shell extract is brand-new.
The invention also discloses walnut shell and be used to prepare the research application that improves treating cardiac and cerebral vascular diseases pharmacologically active control pharmaceutical preparatioies such as blood supply of cardiac muscle, cardiac activity and effect for reducing blood fat.
A further object of the present invention is that providing several is the prescription and the preparation formulation of the pharmaceutical composition of active component with the walnut shell extract.
Walnut shell extract treatment effective dose of the present invention is 50～500mg; Be preferably 100～350mg; Best 150～300mg.
Walnut shell extract pharmaceutical composition of the present invention, it is by walnut shell water or ethanol and total extract thereof and the various pharmaceutical adjuncts pharmaceutically used, is used for medical purpose oral formulations by what reasonable combination was made.
Described drug combination preparation comprises oral formulations such as tablet, capsule, powder, granule and drop pill.Described pharmaceutically acceptable carrier, comprise diluent, filler (as lactose, Polyethylene Glycol) conventional in the preparation, binding agent (starch, microcrystalline Cellulose), disintegrating agent (as carboxymethyl cellulose, the low hydroxypropyl cellulose that replaces), lubricant (as Pulvis Talci, magnesium stearate), wetting agent (as propylene glycol, ethanol), stabilizing agent (EDTA-2Na, sodium thiosulfate, sodium pyrosulfite, sodium sulfite, ethanolamine, sodium bicarbonate, nicotiamide) or the like.Above-mentioned adjuvant can be a common dose, mixes with walnut shell extract with proportioning commonly used, and after the walnut shell extract consumption was determined, the proportioning between each pharmaceutic adjuvant can suitably be regulated as required.
Three, summary of the invention:
Summary of the invention comprises the following aspects:
1. Study on extraction: adopt water and Different concentrations of alcohol to reflux and extract, obtain alcohol extract.
2. studies on acute toxicity: use 30 of the ICR mices of 23-25 gram, male and female half and half experimentize.
3. heart tonifying pharmacology activity research: carry out the research of cardiac activity with isolated rat heart, 6 every group.
3. improve myocardial ischemia effect research: observe with rat integral experiment and improve the myocardial ischemia activity.
4. the research of hypolipidemic activity: use rat hyperlipidemia membranous type to carry out hypolipidemic activity research.
5. the dosage form research of oral formulations: comprise the research preparation of tablet, capsule, powder, granule and drop pill.
(1), extraction and preparation technique research:
The Semen Juglandis leftover bits and pieces walnut shell that shells, shifts operation such as drying and obtains the walnut shell activity extract, extraction ratio 5-9% selected, crushing screening, reflux, extract,, recovery solvent.Solvent for use is water and Different concentrations of alcohol.
(2), acute toxicity testing:
The acute toxicity testing of walnut shell water alcohol method total extract: 10 of ICR mices/group, all do not have death when oral 2000mg/kg, 4000mg/kg, 6000mg/kg, and diet, spirit are all normal.And its optimum effective dose is 50～500mg/kg, and visible medicine is nontoxic substantially, very safe.
(3), heart tonifying pharmacology activity research:
Walnut shell extract ht1, ht2 are to the isolated heart effect
Method:
6 every group of Wistar rats, 12% chloral hydrate 360mg/kg, ip.10mg/kg carries out the whole body heparinization through sublingual vein injection 5mg/ml heparin.Open breast, take out heart, put logical 95%O immediately
2+ 5%CO
24 ℃ of K-H liquid in make asystole, find aorta, the perfusion device mouthpiece is inserted aorta, put then in the Langendorff perfusion device, open logical 95%O
2+ 5%CO
2K-H liquid perfusate carry out coronary artery reverse perfusion (perfusion liquid level and aortic orifice at a distance of 40cm).
Sew up non-traumatic sewing thread and connect tonotransducer through universal pulley at apex of the heart place, after the transducer displacement signal amplifies, lead bio signal acquisition system through MP-150 more digital signal is imported computer, gather and analyze heart rate (HR, bpm), cardiac contractile force (T, g power) and heart contraction rate of change (± dT/dt, g power/s).Behind the balance 30min, what give variable concentrations respectively is subjected to the reagent thing, observes the variation of every index.
The result: (ht1 is the water alcohol method total extract; And ht2 is a 70-100% ethanol extraction part)
Ht1 2.4,8.0mg/L can make cardiac contractile force increase, and reach as high as 24%, and the heart contraction rate of change also increases.Ht2 0.75,2.5mg/L can make cardiac contractile force increase, and reach as high as 59%, and the heart contraction rate of change also increases.The results are shown in Table 1.As seen the activity of effective site (ht2) extract will obviously be better than total extract.
Table 1 walnut shell extract ht1, ht2 to the isolated heart effect (x ± s, n=6)
Annotate: * P＜0.05, * * P＜0.01, the * * * P＜positive contrast medicine of 0.001 vs control.Iso---isoproterenol.
(4), improve the blood supply of cardiac muscle activity research:
Walnut shell extract is to the experimental study of Acute Myocardial Ischemia in Rats effect
1. test objective: observe the protective effect of walnut shell extract, in the hope of providing test basis for clinical practice to Acute Myocardial Ischemia in Rats.
2. medicine and reagent: walnut shell extract (hereinafter to be referred as HT): the total extract of water containing ethanol extraction method, lot number: 07012.With 1%CMC be mixed with 6,20,60mg/mL variable concentrations solution uses for rat oral gavage.Sorbide nitrate (Isosorbide Dinitrate, ID): 5mg/ sheet, 100 every bottle.Tianjin Pacific Pharmaceutical Co., Ltd. produces, lot number: 060402.The suspension that is mixed with 4mg/mL with 1%CMC is used for rat oral gavage.
3. animal and instrument: the Wistar rat, the SPF level, Institute of Radiation Medicine, Chinese Academy of Medical Sciences provides, the animal quality certification number: SCX Tianjin 2005-0001.MP150 leads bio signal collection and analytical system more, Biopac company product.
4. experimental technique:
78 of Wistar rats, male and female half and half, body weight 280.6 ± 8.9 (266～297) g is divided into 6 groups at random by body weight, 10 every group.Give 1%CMC (sham-operation contrast, model control group), walnut shell 70-100% ethanol extraction 30,100,300mg/kg (being subjected to reagent thing group), isosorbide mononitrate 20mg/kg (positive drug group) respectively.The administration volume is the 5ml/kg body weight, every day 1 time, successive administration 5 days.
1h after the last administration, behind the animal via ether general anesthesia, measure normal II and lead the left chest unhairing in electrocardiogram (making normal control) back, conventional iodine tincture, alcohol disinfecting, with left clavicle median line and the 4th rib intersection point is the center, cut skin along the clavicle median line, the flesh layer, circular layer is made purse string suture, get lines crossed to be equipped with and prick, cut off the 4th rib, heart is pulled out the thoracic cavity, under the left auricle and between the pulmonary conus with annular hook, sentence No. 6/0 not damaged suture silk ligation arteria coronaria left anterior descending branch (method is seen accompanying drawing) apart from branch of coronary artery initial part 1.5mm, sham operated rats is not done ligation, and heart is put back to the thoracic cavity, extracts air in the thoracic cavity out, and do the pocket ligation of flesh layer, skin suture; With after lumbar injection 12% chloral hydrate 360mg/kg body weight anesthetized animal, measure behind the ligation arteria coronaria 5,10,15,20,30,45,60,90,120min II leads electrocardiogram, and put cage and raise.
5. result of the test:
5.1 walnut shell extract is to the influence of rat II lead electrocardiogram ST section: after the ligation of rat branch of coronary artery, II leads ECG ST section and obviously raises, with the Sham-operated control group comparing difference significance meaning (P＜0.001) is arranged, present tangible ischemia injury performance, the ST section is raised and be may persist to 2h.Rat orally give walnut shell extract 30mg/kg, every day 1 time, after continuous 5 days, the ST section of 5～20min ECG is significantly reduced, 100,300mg/kg can make the ST section of 5～60min ECG II significantly reduce, and with the model control group comparing difference significance meaning is arranged.The results are shown in Table 1.
5.2 walnut shell extract is to the influence of rat heart rate (HR): rat orally give walnut shell extract 30,100,300mg/kg, every day 1 time, after continuous 5 days, do not have obvious influence to heart rate.The results are shown in Table 2.
Table 1. walnut shell extract to rat ECG-ST section influence (mV, x ± s, n=10)
Annotate: 1. compare * * * P＜0.001 with the sham operated rats non-paired t test; 2. with model group non-paired t test comparison+P＜0.05, ++ P＜0.01, +++P＜0.001.
Table 2. walnut shell extract to rat HR influence (bpm, x ± s, n=10)
Annotate: 1. compare P＞0.05 with the sham operated rats non-paired t test; 2. compare P＞0.05 with the model group non-paired t test.
6. conclusion:
The acute myocardial ischemia model that adopts rat branch of coronary artery ligation method to cause, the function of resisting myocardial ischemia of observation walnut shell extract.The result shows, rat orally give walnut shell extract 30mg/kg, every day 1 time, after continuous 5 days, the ST section of 5～20min ECG is significantly reduced, 100,300mg/kg can make the ST section of 5～60min ECG II significantly reduce, and with the model control group comparing difference significance meaning arranged.Rat orally give walnut shell extract 30,100,300mg/kg, after continuous 5 days, do not have obvious influence to heart rate at every day 1 time.The The above results prompting, the orally give walnut shell extract has the obvious treatment effect to Acute Myocardial Ischemia in Rats.
(5), the research of hypolipidemic activity:
Get 60 of rats, grouping, oral administration, continuous 30d, simultaneously except that negative control group feeding normal feedstuff, all the other 4 groups of feeding high lipid foods (cholesterol 3%, Adeps Sus domestica 10%, yolk powder 5%, methylthiouracil 0.3%, normal feedstuff 81.7%, adding suitable quantity of water after fully mixing thoroughly, to pinch into fritter edible).Get the preceding fasting 12h of blood, breaked end respectively 6 in the 15th and 30 day and get blood, measure serum cholesterol (TC), triglyceride (TG) and high density lipoprotein (HDL-C) content.The result shows: the administration group can obviously reduce rat blood serum hypercholesterolemia (TC) and the triglyceride (TG) that high lipid food brings out, the content of high density lipoprotein increasing (HDL-C).The results are shown in Table 3.Show that medicine has obvious blood fat reducing and prevents and treats arteriosclerotic effect.
Table 3 is tried before the rat experiment and experiment the 15th, 30dTC, TG and HDL-C content (mmol/L, X ± s)
Annotate: compare * P＜0.05 with the high blood lipid model group; * P＜0.01.
(6), the research of medicament form of pharmaceutical preparation:
(1) tablet: different content, coating: the extract tabletting is made the use of tablet, and the adjuvant that is used as excipient has MgSO4, corn starch, Pulvis Talci (specification: 100mg, 150mg, 200mg, 300mg).
(2) capsule: common and enteric coated capsule, different size.
(3) powder: after adding certain adjuvant after the extract drugs, dry levigation.
(4) dissolved granule: packed granulated quickly dissolving.
(5) drop pill: increase absorbance and infiltration rate, rapid-action.
The preparation concrete operations of various pharmaceutical preparatioies are as follows:
(1). take by weighing and contain 10～70% walnut shell extracts by the configuration total amount, the filler of adding 30～90%, binding agent, disintegrating agent, lubricant, correctives etc., fully mix and make granule, 60～70 ℃ of dryings after 2～4 hours, compacting makes every to contain walnut shell extract 50～150mg in flakes.
(2). maybe the granule of making directly is filled in the various hungry area softgel shells, makes every capsules contain walnut shell extract 50～200mg, be capsule.Certainly each preparation proportioning and adjuvant are selected difference to some extent for use.
(3). take by weighing and contain 20～70% walnut shell extracts by the configuration total amount, add cosolvent such as dextrin, the correctives etc. of 30-80%, dissolve even after drying, the pulverized powder pack promptly gets powder.
(4). take by weighing and contain 10%～60% walnut shell extract by the configuration total amount, add 40%～90% filler, disintegrating agent, correctives etc., fully mix with 12～14 mesh sieves and make granule.In the fully back pack of 60～70 ℃ of dryings, make every gram granule contain walnut shell extract 100～500mg.
(5). take by weighing and contain 10%～60% walnut shell extract by the configuration total amount, the cosolvent Polyethylene Glycol of adding 40%～90%, antioxidant etc., heating and melting temperature (40～90 ℃) makes raw material and adjuvant fully miscible, drip and make ball, make every drop pill contain walnut shell extract 5～20mg.
Formulation rate of the present invention can be according to all multifactor adjustment such as route of administration, patient age, body weight, disease type and the orders of severity, and daily dose is for being generally 50～500mg; Be preferably 100～250mg; Best 150～200mg.Can show suitable change according to clinical case.
Four, the specific embodiment:
The present invention will be further described below by embodiment, but be not to be limitation of the present invention:
Embodiment 1:
1.500g walnut shell was pulverized 20 mesh sieves, added 70% alcohol reflux 4 times of 1200～1500ml, refluxed 1 hour at every turn.Merge ethanol liquid rotary evaporation, 60 ℃ are reclaimed ethanol, shift and put into 60 ℃ of oven dried, obtain 32.6 gram brownish red extracts, extraction ratio 6.5%.
2.2000g walnut shell was pulverized 20 mesh sieves, added the dehydrated alcohol reflux, extract, 4 times of 10000～12000ml, refluxed 1 hour at every turn.Merge ethanol liquid rotary evaporation, 60 ℃ are reclaimed ethanol, shift and put into 60 ℃ of oven dried, obtain 120.5 gram brownish red extracts, extraction ratio 6.1%.
3.2000g walnut shell was pulverized 20 mesh sieves, added 95% alcohol reflux 3 times of 10000～12000ml, refluxed 1 hour at every turn.Defective material refluxed 1 hour with the 10000ml distilled water again, extracted 2 times, merged water alcohol liquid rotary evaporation, and 60 ℃ are reclaimed ethanol, shift and put into 60 ℃ of oven dried, obtained 172.0 gram brownish red extracts, extraction ratio 8.6%.
Embodiment 2:
Get walnut shell extract 500g, medical starch 300g, dextrin 300g, 50% ethanol is an amount of, above-mentioned raw materials is fully mixed make granule, 60～70 ℃ of dryings 2～4 hours, is pressed into 10000, sugar coating, every contains walnut shell extract 0.05g.
Embodiment 3:
Get walnut shell extract 500g, medical starch 500g, 50% ethanol is an amount of, granulates granulate, oven dry, dress 2# capsule, every 0.1g.
Embodiment 4:
Get walnut shell extract 500g, medical starch 300g, dextrin 300g, 50% ethanol is an amount of, above-mentioned raw materials is fully mixed make granule, and 60～70 ℃ of dryings 2～4 hours, pack was sealed, and is dissolved granule.
Embodiment 5:
Get walnut shell extract 40g, Polyethylene Glycol-4000 20g, Polyethylene Glycol-6000 60g, sodium pyrosulfite is an amount of, and heating and melting temperature (40～90 ℃) makes raw material and adjuvant fully miscible, drip and make 20000 drop pill, make every drop pill contain walnut shell extract 3.0～5.0mg.
Although the present invention has done detailed description in conjunction with its special embodiment, clearly concerning the personage that the present technique field is familiar with, still can make various changes and improvements, but can not depart from spirit of the present invention and protection domain.
Claims (5)
1. process: a kind of process with water or ethanol extraction walnut shell extract, this technology comprises, remove the walnut shell leftover bits and pieces that removes Semen Juglandis, through pulverizing, sieve, extract recovery solvent and oven dry, porphyrize technology, obtain being rich in its glycosides of Semen Juglandis quinone, tannin, polysaccharide and vegetable protein isoreactivity composition.
2. one kind is the pharmaceutical composition of active component with walnut shell extraction effective site, and said composition contains the activity extract for the treatment of effective dose, and one or more pharmaceutically acceptable carrier.The oral formulations of making has tablet, powder, capsule, and drop pill.
3. walnut shell extract is used to the application for preparing cardiac drug, improve myocardial ischemia drug.
4. the walnut shell effective part extract is prevented and treated the application of medicine aspect arteriosclerosis and the cardiovascular and cerebrovascular disease at blood fat reducing, blood pressure regulation.
5. with other organic solvent extraction, or the application that is used for heart tonifying and cardiovascular medicament formulation art of the walnut shell effective active composition that obtains by other separating technology means or position.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101982187A (en) * | 2010-10-27 | 2011-03-02 | 大理学院 | Walnut shell extract and anti-HIV pharmaceutical application thereof |
| CN103340923A (en) * | 2013-06-20 | 2013-10-09 | 吕梁学院 | Preparation method for walnut tannin |
| CN105418693A (en) * | 2015-10-31 | 2016-03-23 | 中北大学 | Method for extracting tannin from walnut shells and application of obtained tannin |
| CN109777741A (en) * | 2019-01-15 | 2019-05-21 | 昆明理工大学 | A kind of method that walnut shell efficiently utilizes |
-
2007
- 2007-03-26 CN CNA2007100569968A patent/CN101274011A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101982187A (en) * | 2010-10-27 | 2011-03-02 | 大理学院 | Walnut shell extract and anti-HIV pharmaceutical application thereof |
| CN101982187B (en) * | 2010-10-27 | 2012-08-22 | 大理学院 | Walnut shell extract and anti-HIV pharmaceutical application thereof |
| CN103340923A (en) * | 2013-06-20 | 2013-10-09 | 吕梁学院 | Preparation method for walnut tannin |
| CN103340923B (en) * | 2013-06-20 | 2015-02-18 | 吕梁学院 | Preparation method for walnut tannin |
| CN105418693A (en) * | 2015-10-31 | 2016-03-23 | 中北大学 | Method for extracting tannin from walnut shells and application of obtained tannin |
| CN105418693B (en) * | 2015-10-31 | 2018-03-02 | 中北大学 | The application of the extracting method of tannin and its gained tannin in a kind of walnut shell |
| CN109777741A (en) * | 2019-01-15 | 2019-05-21 | 昆明理工大学 | A kind of method that walnut shell efficiently utilizes |
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