CN102526182A - Application of Prunella vulgaris Linn. and extract of Prunella vulgaris Linn. to preparation of product for inhibiting angiogenesis - Google Patents
Application of Prunella vulgaris Linn. and extract of Prunella vulgaris Linn. to preparation of product for inhibiting angiogenesis Download PDFInfo
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- CN102526182A CN102526182A CN201010610675XA CN201010610675A CN102526182A CN 102526182 A CN102526182 A CN 102526182A CN 201010610675X A CN201010610675X A CN 201010610675XA CN 201010610675 A CN201010610675 A CN 201010610675A CN 102526182 A CN102526182 A CN 102526182A
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Abstract
The invention discloses novel application of Prunella vulgaris Linn. or an extract thereof. The novel application of the Prunella vulgaris Linn. or the extract thereof is angiogenesis inhibition; and the Prunella vulgaris Linn. or the extract thereof has a broad application prospect in the treatment of vascular dysplasia diseases such as rheumatoid disease, tumor, retina eyeground disease, psoriasis and the like. The invention provides a novel content and a novel direction for the development, application and research of the medicine, and also provides a novel means for the research on the treatment of vascular dysplasia diseases by Chinese medicines.
Description
Technical field
The present invention relates to the new purposes that Spica Prunellae and extract thereof suppress angiogenic growth (angiogenesis), particularly Spica Prunellae and extract thereof suppress the application in the angiogenesis product in preparation.
Background technology
Angiogenesis is meant the process that is formed new blood capillary by former already present blood capillary through sprouting.It is not only the basis of physiological process such as body normal development, reproduction and tissue repair, also in multiple pathological process, plays an important role.If angiogenesis is excessive, relevant with diseases such as rheumatoid, tumor, retina retinopathy and psoriasiss, angiogenesis inhibitors potential and widely purposes make it become current medicament research and development focus.(Judah?Folkman.Angiogenesis：an?organizing?principle?for?drug?discovery?Drug?Discovery，2007，6(4)：273-286.)
Spica Prunellae (Prunella vulgaris Linn.) is Chinese medicine commonly used; Beginning is stated from Shennong's Herbal; Bitter in the mouth cold in nature, having relieves inflammation or internal heat makes eye bright, the effect of mass dissipating and swelling eliminating, and ancient Chinese medicine doctor is used for treating diseases such as pestilence, acute mastitis, ophthalmalgia, jaundice, gonorrhea and hypertension more.Modern pharmacology research proof Spica Prunellae has pharmacological action widely; As have effects such as blood pressure lowering, blood sugar lowering, antibiotic, antiinflammatory, antiallergic, antioxidation, removing free radical and antiviral; External Spica Prunellae is as a kind of self-rehabilitation medicine, has sore throat in alleviation, aspects such as heating and accelerating wound healing are widely used and come into one's own.
Environment was similar with human body in screening angiogenic activity material was capable of using; And the animal model that angiogenic process is easy to observe detects; Wherein (chorioallantoie membrane CAM) is the ideal body inner model of research vascularization and anti-angiogenic formation to chick chorioallantoic membrane.Angiogenesis mainly occurs in the embryo development procedure; It is the most active that Embryo Gallus domesticus is hatched 3-5 days, 7-10 days angiogenic growth, and CAM blood vessel atrophy gradually after 12 days is the medicine irritation Best Times so choose 8-12 days; Utilize stereoscopic microscope, observe and calculate the quantity of blood vessel.That this technology not only can be used for is qualitative, angiogenesis in the quantitative study body, also have experiment material be easy to get, easy and simple to handle, experimental period short, do not need special installation, result to be easy to observe and suitable large sample advantage such as repeats and is used widely.The earliest CAM is conducted a research as neoplasm metastasis and erosion models and also just be based on its abundant and angiogenic growth characteristic (Leighton, J.Invasion and metastasis of heterologous tumors in the chick embryo.In " Progress.Experimental Tumor Research " 1964 intuitively; 14:98.).The CAM model effect that is used to estimate new drug has at present obtained approval (the Vargas A of U.S. food and drug administration; Zeisser-Labouebe M; Lange N; Et al.The chick embryo and its chorioallantoic membrane (CAM) for the in vivo evaluation of drug delivery systems.Adv Drug Deliv Rev, 2007,59 (11): 1162-1176.).
Based on of influence and the effect of CAM model evaluation Chinese medicine to angiogenic activity, not only can further excavate the value of Chinese medicine, can also the treatment of angiogenic disease be prolonged human longevity and make contributions.
Summary of the invention
The new purposes that the purpose of this invention is to provide Spica Prunellae and extract thereof.Show that preparation suppresses to use separately in medicine and the health product of angiogenesis or with other drug and the use of effective ingredient compatibility.
Above-mentioned Spica Prunellae extract can be Spica Prunellae water or alcohol extract.The Spica Prunellae water extract also can prepare according to conventional method; Obtain like available following method: choose medical material Spica Prunellae (flower, fruit ear and the herb of labiate Spica Prunellae Prunella vulgaris Linn.) and add 5～10 times of pure water; Heating; Boil the back and keep slight boiling condition to continue heating 0.5～1 hour, filter then and collect filtrating 1; Then in filtering residue, add 5～10 times of pure water again, heating is boiled the back and is kept slight boiling condition to continue heating 0.5～1 hour, filters then and collects filtrating 2; To filtrate 1 with filtrating 2 mixings, drying obtains the Spica Prunellae water extract.
Spica Prunellae ethanol extract can prepare according to conventional method; Obtain like available following method: the Spica Prunellae of pulverizing and dehydrated alcohol or 50% ethanol mix with 1: 10 (volume fraction); Boiling water backflow 5h collects extracting solution, residue boiling water backflow 3h once more after the coarse filtration; Merge 2 times extracting solution, rotary evaporation obtains Spica Prunellae alcohol extraction concentrated solution.
Medicine with Spica Prunellae or extract preparation can be processed various ways such as tablet, capsule, oral liquid, powder, granule, unguentum, cream, injection.The medicine of above-mentioned various dosage forms all can be according to the conventional method preparation of pharmaceutical field.
Health product with Spica Prunellae or extract preparation can be processed various ways such as tablet, capsule, oral liquid, powder, granule, unguentum, cream.The medicine of above-mentioned various dosage forms all can be according to the conventional method preparation of pharmaceutical field.
The present invention adopts this model organism of CAM, as detection means, adopts method of serum pharmacology to observe the influence of Spica Prunellae to angiogenesis with the whole animal experiment; Find that Spica Prunellae can significantly suppress angiogenesis; Drug action is more obvious than Western medicine thalidomide, and the CAM visual field is no intensive angiogenesis down, fails to see blood-vessels mesh network clearly; For the study on mechanism of this medicine provides new content and direction, also new thinking and means are provided for tcm development utilization research.
Description of drawings
Fig. 1 is that Spica Prunellae extract and thalidomide suppress CAM angiogenesis figure.
Fig. 2. be Spica Prunellae extract and thalidomide result to the blood vessel number affects
Experimentation, Spica Prunellae suppress the test of chick chorioallantoic membrane angiogenesis
1 material
1.1 Embryo Gallus domesticus
Purchase in kind of chicken or chicken farm.
1.2 medicine and reagent
The Spica Prunellae water extract prepares according to following method: choose twice of medical material Spica Prunellae (labiate Spica Prunellae Prunella vulgaris Linn. flower, fruit ear and herb) decocting; Decocting liquid filters; Mix post-heating and be concentrated into 1g/mL, this is the Spica Prunellae water extract.
Spica Prunellae ethanol extract prepares according to following method: the Spica Prunellae of pulverizing and dehydrated alcohol or 50% ethanol mix with 1: 10 (volume fraction); Boiling water backflow 5h; Collect extracting solution; Residue boiling water backflow 3h once more merges 2 times extracting solution after the coarse filtration, and rotary evaporation obtains Spica Prunellae alcohol extraction concentrated solution 1g/mL.
Thalidomide (the luxuriant imidodicarbonic diamide of chemical name α-Phthalide, common name Thalidomide, neurosedyn.Sigma company).
Mixed cellulose ester microporous membrane (0.45 μ m, the inferior scavenging material in last Haixing County factory); Qualitative filter paper (the extraordinary paper plant in Fuyang, Hangzhou); All the other reagent are homemade.
1.3 instrument
MIR-553 constant incubator (Japanese SANYO company); S6E stereomicroscope (German Leica company); XTS30-D stereomicroscope (Tyke, Beijing Instr Ltd.), desk centrifuge (go up Hai'an booth company), MLS-3780 high-pressure sterilizing pot (Japanese SANYO company) etc.
2 methods
2.1 test is divided into groups and the pastille serum preparation
Male mice in kunming is divided into Spica Prunellae water extract group, Spica Prunellae dehydrated alcohol extraction thing group, Spica Prunellae 50% ethanol extraction group, Western medicine thalidomide positive controls and normal saline matched group at random; Two drug group routines are irritated stomach by body weight and are given and the medicine that is equivalent to 5 times of people's clinical medicine doses, and the normal saline group adopts isodose normal saline, 1 time on the one; 1h after thing is taken medicine in last 1 filling of 5d; Pluck eyeball and get blood, the about 0.5mL of centrifugal preparation serum places sterile test tube; 56 ℃ of deactivation 30min seal in-20 ℃ of refrigerators frozen.
2.2 Embryo Gallus domesticus is cultivated
Hatching egg embathes with 1 ‰ bromo geramine aqueous solutions to be wiped driedly, and air chamber places the hatching of (37 ± 0.5) ℃ calorstat to cultivate up, and built-in wet dish keeps relative humidity 65%, and every day, egg-turning was 2 times, observed the hatching egg growing state down in the transmission lamp behind the 3d, rejected unfertilized egg.
2.3 chick chorioallantoic membrane (CAM) experiment
Get the 7d instar chicken embryo, random packet, iodophor disinfection egg plenum surface with the window of emery wheel and the about 1cm2 of dissecting needle breakdown, exposes CAM, and aseptic tape seal continues hatching.The next day; The directly about 0.5cm of cut-off; Warp autoclaved cellulose mixture microporous filter membrane sheet in advance places on the CAM as carrier, and the pastille serum 20 μ L that drip different groups respectively seal continued and hatch 72h on carrier; Adding fixedly 15min of 1: 1 methanol and acetone mixed liquor room temperature, is that the about 3cm of CAM is cut at the center with the carrier
2, open up film on filter paper, airing.
2.4 the result observes
With filter paper is background, checks carrier blood vessel number on every side under the stereomicroscope, respectively organizes the angiogenesis situation.
2.5 statistical analysis
Adopt the ANOVA variance test to analyze the life experiment result, quantitative data is represented with
; Adopt blood vessel quantity variance between two sample t method of inspection comparable group.
3 results
3.1CAM the angiogenesis form of expression
Fig. 1. the microscopy result of drug influence angiogenesis
Visible by Fig. 1; The angiogenesis of CAM naturalness is the vein shape; The administration group presents the vasoganglion of radial growth in various degree around carrier, the blood vessel quantity in its Chinese medicine and western medicine thalidomide group, Spica Prunellae ethanol extract group and the Spica Prunellae 50% alcohol extract group visual field is less than the normal saline matched group, but still can see blood vessel network clearly; And Spica Prunellae water extract group does not only have intensive angiogenesis, fails to see blood-vessels mesh network clearly under the visual field.
3.2 vascular counts
Table 1. respectively organize angiogenic growth quantity (X ± SD, n=9)
Group blood vessel quantity
Normal saline group 114.33 ± 17.5
Thalidomide group 71.89 ± 11.72
*
Spica Prunellae water extract group 25.00 ± 7.58
* #
Spica Prunellae ethanol extract group 75.11 ± 8.81
*
Spica Prunellae 50% alcohol extract group 77.89 ± 8.10
*
Compare P＜0.01 with the normal saline group,
#Compare P＜0.01 with the thalidomide group
Above-mentioned experimental result is qualitative from form and quantity two aspects, show that Spica Prunellae extract has the effect of significant inhibition angiogenic growth quantitatively; Wherein Spica Prunellae water extract inhibition effect is more remarkable than Western medicine thalidomide, and the inhibitory action of alcohol extract is suitable with the Western medicine thalidomide.
The specific embodiment
Experimental technique described in the following embodiment is applicable to medicine and health product preparation, like no specified otherwise, is conventional method; Said reagent and biomaterial like no specified otherwise, all can obtain from commercial sources.
The preparation tablets of embodiment 1, Spica Prunellae extract
Spica Prunellae adds 5～10 times of pure water, divides 2 decoctions, merges medicinal liquid and is concentrated into medicinal liquid than 1: 1, adds the auxiliary materials and mixing of 100 mesh sieves, selects lactose for use; Starch, dextrin are diluent, and CMS-Na is a disintegrating agent, and ethanol is wetting agent, and adding 1% magnesium stearate is lubricant; Prepare suitable soft material, granulate 80 ℃ of dryings, granulate, last machine tabletting through 16 eye mesh screens; The bag film-coat gets product, and average sheet heavily is about every 0.29g, and dosage is every 0.125g.
The capsule preparation of embodiment 2, Spica Prunellae extract
Spica Prunellae water extract or ethanol extract added the auxiliary materials and mixing of 100 mesh sieves, selected lactose for use, and starch, dextrin are diluent; Ethanol is wetting agent, pushes 40 mesh sieves and granulates, and in 60 ℃ of dryings, granulate sieves; Pour into No. 1 capsule, make finished product, dosage is every capsule 0.125g, the heavily about 0.3g of every capsule.
The oral liquid preparation of embodiment 3, Spica Prunellae extract
Spica Prunellae adds 5～10 times of pure water, divides 2 decoctions, merges 2 times decoction liquor; Be condensed into medicinal liquid than 1: 1, remove tannin, protein, waxiness isocolloid labile element through the clarifier treatment process, post precipitation is got supernatant; Adding purified water to prescribed volume boils; Be distributed into 10ml/ after the cooling and prop up, 100 ℃ of sterilizations got product in 30 minutes, and every dosage is 0.125g.
Among the present invention, Spica Prunellae extract has utmost point significant inhibitory effect to angiogenic growth, points out this medicine and extract thereof the potential value aspect angiogenic disease.
Claims (3)
1. Spica Prunellae or Spica Prunellae extract suppress the application in the angiogenesis product in preparation.
2. application according to claim 1 is characterized in that: said product is medicine or health product.
3. application according to claim 1 and 2 is characterized in that: said Spica Prunellae extract is Spica Prunellae water or alcohol extract.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105477204A (en) * | 2016-01-11 | 2016-04-13 | 高淼淼 | Composition for adjuvant treatment on occlusion of central retinal artery and preparing method of composition |
| CN113975283A (en) * | 2020-07-27 | 2022-01-28 | 韩国韩医药振兴院 | Anti-angiogenic composition comprising sepsis acid A isolated from Prunella vulgaris as active ingredient |
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| WO2004019961A1 (en) * | 2002-08-30 | 2004-03-11 | Biopharmacopae Design International Inc. | Plant extracts for treatment of angiogenesis and metastasis |
| US20070299046A1 (en) * | 2006-06-26 | 2007-12-27 | Mai Nguyen Brooks | Orally available light-independent antineoplastic compounds |
| CN101180069A (en) * | 2005-03-23 | 2008-05-14 | 李氏大药厂（香港）有限公司 | Herbal compositions useful in cancer treatment |
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Patent Citations (4)
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| WO2002013835A1 (en) * | 2000-08-11 | 2002-02-21 | Ashni Naturaceuticals, Inc. | Composition exhibiting synergistic antioxidant activity |
| WO2004019961A1 (en) * | 2002-08-30 | 2004-03-11 | Biopharmacopae Design International Inc. | Plant extracts for treatment of angiogenesis and metastasis |
| CN101180069A (en) * | 2005-03-23 | 2008-05-14 | 李氏大药厂（香港）有限公司 | Herbal compositions useful in cancer treatment |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105477204A (en) * | 2016-01-11 | 2016-04-13 | 高淼淼 | Composition for adjuvant treatment on occlusion of central retinal artery and preparing method of composition |
| CN113975283A (en) * | 2020-07-27 | 2022-01-28 | 韩国韩医药振兴院 | Anti-angiogenic composition comprising sepsis acid A isolated from Prunella vulgaris as active ingredient |
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Application publication date: 20120704 |