CN102688501A - Proanthocyanidin B2 phospholipid compound, and preparation method and application thereof - Google Patents

Abstract

The invention relates to the technical fields of medicines, health products and the like, and provides a proanthocyanidin B2 phospholipid compound associated with effective parts of traditional Chinese medicines, a preparation method thereof and application thereof mainly used for vascular dementia. The proanthocyanidin B2 phospholipid compound is composed of proanthocyanidin B2 and soya bean lecithin, yolk lecithin or synthetic phospholipid according to a mol ratio of (1:10)-(15:1). The preparation method comprises the following steps: heating, refluxing, washing and drying. By putting the proanthocyanidin B2 and a certain amount of phospholipid in an appropriate solvent to form the phospholipid compound, the physical and chemical properties of the proanthocyanidin B2 are changed, the lipid solubility and stability are enhanced, and the transmembrane transport performance is improved, thereby increasing the bioavailability. The phospholipid compound has the advantages of powerful action, obvious effect, stable property, high use safety, abundance of sources, simple preparation process and good reproducibility, and is suitable for industrial production.

Description

The procyanidin B 2 phosphatide complexes

Technical field

The present invention relates to medicine, food, health product and technical field of beverage; Specifically relate to a kind of composition and method of making the same relevant and purposes with effective ingredient in Chinese; More particularly relate to a kind of procyanidin compounds, specifically relate to procyanidin B 2 again and (claim again: phosphatide complexes procyanidin B 2).

Background technology

One, the research overview of procyanidin B 2

1, anthocyanidin

Anthocyanidin (Anthocyanins) is a kind of water colo(u)r, can change color along with the soda acid of Cell sap.It is then red partially that Cell sap is acidity, and it is then blue partially that Cell sap is alkalescence.Anthocyanidin is one of main pigment that constitutes petal and fruit color.Intercrescence becomes approach (flavonoids biosynthetic pathway) to generate with flavonoid via phenylpropionic acid class synthesis path (phenylpropanoid pathway).

The factor that influences the anthocyanidin colour generation comprises the structure of anthocyanidin, pH value, is total to color effect (copigmentation) etc.The peel colour generation receives the influence of inherence, extrinsic factor and cultivation technique.Light can increase anthocyanidin content; High temperature can make the anthocyanidin degraded.Anthocyanidin is plant secondary metabolism product, on physiology, plays the part of important role.The color of petal and fruit can attract animal to pollinate and seed dispersal (Stintzing and Carle, 2004).Reach the epidermis cell and the following table cortex of stem and leaf in the tissue of common what flower, fruit.The part fruit is with the shade decision fruit market price.

Anthocyanidin belongs to the flavonoid class (flavonoids) in the what phenolic compound.Basic structure comprises two phenyl ring, and links (C by the unit of one 3 carbon ₆-C ₃-C ₆).Intercrescence becomes approach to generate to anthocyanidin with flavonoid via the phenylpropionic acid path, by many ferment regulation and control catalysis.With pelargonidin (pelargonidin), cyanidin (cyanidin), delphinidin (delphinidin), peonidin (peonidin), petunidin (petunidin) and six kinds of non-glycoside bodies of enidin (malvidin) (aglycone) is main.Anthocyanidin because of hydroxyl number (OH), methylate (methylation), factor such as candy baseization (glycosylation) number, candy kind and link position presents different colours (model and Qiu, 1998).The performance of color is because of the change of biochemical environmental condition, as receives the influence (Clifford, 2000) of pH value among anthocyanidin concentration, color effect altogether, the vacuole.

Orange and yellow is the effect of carotene.In Radix Dauci Sativae, found beta-carotene in 1910, found other 2 kinds of carotene isomers later on altogether, be respectively: α, β, three kinds of isomers of γ.Beta-carotene in 1958 patent (US2849495, on August 26th, 1958, the patentee: Hoffmann La Roche), mainly from the ocean, extract at present, but also synthetic.

2, procyanidin B 2

Procyanidin compounds (Oligomeric Proantho Cyanidins; Be called for short: OPCs or OPC); Claim to concentrate gene (Pycnjogenols) again; Being a kind of bioflavonoids that special molecular structure is arranged, is cofactor efficiently, be that generally acknowledge in the world at present, active the strongest, remove the most effectively Natural antioxidant of free radical in the human body, remove free radical with and the material of anti-aging effects.Be generally the rufous powder, feeble QI, puckery, water-soluble and most organic solvent.Be generally Semen Vitis viniferae extract or Pinus pinaste bark extract.

Procyanidin compounds is a kind of new and effective antioxidant, and the free radical scavenger of imitating the most by force that is so far to be found has very strong activity in vivo.The free radical resisting oxidability that experiment showed, OPCs is 50 times of vitamin E, ascorbic 20 times, and absorb rapidly fully, can reach the highest haemoconcentration in oral 20 minutes, metabolic half life is for 7 hours.

Procyanidin compounds is rich in the plant flavonoids material (flavonoids) the most useful to human body---proanthocyanidin glycosides (Proanthocyanidin) or title procyanidin (Procyanidin or Procyanidins are called for short PC).

Procyanidin is present in the plant widely; Be a kind of by procyanidin monomers (mainly being catechin, epicatechin etc.), Oligomeric Proanthocyanidins (monomeric two, three, four polymeric acceptors; English is Oligomeric procyanidins; Be called for short: (more than the pentamer, English is Procyanidolic polymers, is called for short: the chemical compound of PPC) forming PCO) to gather procyanidin with height.The most basic procyanidin monomers, a kind of is catechin, and another kind is an epicatechin, and these two kinds of monomers can form oligomer or polymer through polymerization, usually two～tetramer are called oligomer by the size of the degree of polymerization, then are called high polymer more than the pentamer.

Wherein, dimer distributes the widest in all kinds of procyanidins, and antioxidant activity is the strongest, is most important one type.Dimer has multiple isomer because of 2 monomeric conformations or condensation key position difference, 8 kinds of isomers at present having identified, and procyanidin B 2 is an active the strongest dimer in 8 kinds of isomers.Its structural formula is following:

But because the procyanidin water solublity is strong, the transmembrane transport poor performance is difficult to be absorbed the entering body through gastrointestinal wall and circulates; And in gastrointestinal tract character unstable, be prone to be degraded, cause bioavailability very low.Influence the performance of procyanidin B 2 oral formulations curative effect, limited its use clinically.

3, source

Procyanidin compounds is generally Semen Vitis viniferae extract.Though procyanidin compounds has the effect of mystery like this, human body can't produce procyanidin compounds, and procyanidin compounds focuses mostly at the skin of plant, shell, seed, leaf, the first-class position of bar, like Semen Vitis viniferae, skin etc.; And best resource is picked up from fruit nymphaeaceous and leaf, can be higher than (that extracts in general Semen Vitis viniferae, the skin can only reach 90% at most) more than 98% with the OPC purity of Nelumbonaceae plant extract.

In recent years; The countries in the world scholar has carried out number of research projects to physicochemical property and the biological nature of OPCs, and makes a breakthrough, in Semen Vitis viniferae extract; OPCs has higher content, and the content height of oligomer has guaranteed the high curative effect of the procyanidin compounds of unit quantity.And the bioflavonoids OPCs that from Semen Vitis viniferae, extracts compares with most of flavone, and structure is special, good water solubility, effectiveness is high, bioavailability very easily is absorbed by the body more than 90%, be find up to now imitate natural antioxidant the most by force.

According to the achievement in research of 1951 ~ 1971 years and 1972 ~ 1978 years Jacques Masquelier, Semen Vitis viniferae is the source of the OPCs that is considered to best, is rich in 92 ~ 95% OPCs.Simultaneously the scientific research result shows, grape seed extract OPC s is following important defying age and anticancer natural product, and Semen Vitis viniferae extract obtains medical licence in France.

In present market, the OPCs all size emerges, and only has monomer more than 10%; Procyanidin is more than 95%, and polyphenol is only real grape seed extract OPC s more than 90%, if monomer is lower than 3%; Polyphenol 60 ~ 70% even reach 90% is generally then thought procyanidin PPC.

It should be noted that skin, shell, seed, leaf, the processing of bar un-extracted of occurring in nature, OPC wherein can not extract out, and natural human body can not absorb.

4, the various health-care of procyanidin compounds

Theoretical research through for many years proves that procyanidin compounds has plurality of health care functions, comprising:

1. protect the blood circulation of human bodies through three approach: remove the oxygen-derived free radicals that exists in the blood; Help body to stop the generation of oxygen-derived free radicals; Strengthen the integrity of blood vessel wall.

2. prevent collagenase and elastoser Degradation, thereby help keeping the elasticity of skin, the effect of performance anti aging effect connective tissue.

3. can strengthen the microcirculation of optical fundus retinal capillary and extremity tip blood capillary.

4. health and the microcirculation improvement through keeping blood vessel makes each organ and tissue obtain more competent blood supply.

5. can raise immunity, show that anthocyanidin improves the immunne response ability of the white mice cytokine 2 of the white mice that feeds ethanol and retroviral infection.

6. procyanidin compounds also can strengthen ascorbic effect, prolongs the time that effect takes place in human body vitamin C.

7. the dissolubility of procyanidin compounds in water is very good, and oral post-absorption is good, and be rapid-action in the entering human body, and action time is lasting.

8. as health food, safety is extremely important.The result of multiple zoopery and experimental system research proves that nontoxic, the no mutagenicity of procyanidin compounds, non-carcinogenesis, nothing cause birth defect, no sensitization, so anthocyanidin is safe.The concrete elaboration as follows:

(1) blood circulation

In Europe, in order to improve blood circulation, treat diabetic retinopathy, alleviate edema and to suppress varicosis etc., oneself is used for clinical treatment decades anthocyanidin.Anthocyanidin can be strengthened blood capillary, tremulous pulse and vein blood vessel, and therefore, it has the effect of detumescenceization silt.The resistance of blood capillary reduces and permeability improves, and makes cell absorb nutrient and eliminating refuse more easily.This is the function of blood circulation with transporting refuse to carry nutrient.Heart is responsible for taking out pressure blood; Tremulous pulse and vein blood vessel pumping blood; Give cell and be responsible for transporting nutrition, what transport refuse again is blood capillary.Anthocyanidin can the scavenger cell film in water solublity and fat-soluble free radical, therefore, suppressed to discharge the process that some enzyme removes to injure capillary wall.

The nourishing effects of anthocyanidin can be observed in a short period of time.Professor HenriChoussat of Dogue de Bordeaux university has done single test, and the experimenter has 47 people, and the age, everyone took 100 milligrams of anthocyanidin from 37 years old to 85 years old.Find after 27 hours that the resistance of blood capillary has reduced 40%.

(2) vision protection

Diabetic retinopathy is the sign of diabetes, and it is little hemorrhage the causing of eyes blood capillary, be grow up the blind person commonly encountered diseases because of.France allow with the anthocyanidin treatment should disease a lot of year.This method has significantly reduced the eyes capillary hemorrhage, has improved vision.Anthocyanidin also has been used for preventing the complication behind the diabetics cataract operation.

1998, brainstrust selected many people that do not have the oculopathy eye to hinder to do experiment, and investigates anthocyanidin and whether can alleviate nyctalopia.The participant is divided into two groups, and one group is the driver who drives evening, and one group is all day and the people of computer screen for accompanying.After four weeks, reexamine their anti-blind ability, 98% experimenter has had improvement.

(3) edema

Edema is that moisture, electrolyte in the blood etc. has infiltered tissue and causes its normally injury swelling.Sit to such an extent that healthy subjects of a specified duration excessively has edema, woman month is through before having edema, and the injured edema that often causes of moving has edema after some operation, and some diseases also can cause edema.

Research shows, takes anthocyanidin every day one time, and edema can obviously be alleviated.Clinical trial was done by Dogue de Bordeaux university, and having investigated the mean age is 60 years old, and 40 patients of edema disease are arranged.The participant is divided into two groups, the patient be in 60 days every day take 300 milligrams anthocyanidin, proprietary disease alleviates.After 30 days, 26% patient's swelling disappears.After 60 days, the recovery from illness of the edema of experimenter's shank of 63%.

(4) skin care

People from Europe claims that anthocyanidin is the youth nutriment, skin vitamin, oral cosmetics.Because it can recover the collagen protein vigor, make skin level and smooth and flexible.Collagen protein is the fundamental component of skin, and is that a kind of our health that makes becomes as a whole colloid substance.Vitamin C is the necessary nutriment of biochemical synthetic collagen protein.Anthocyanidin comes into force more vitamin C, this means, vitamin C can more easily go to accomplish its all functions (comprising that collagen protein produces).Anthocyanidin is connected on the collagen protein, can stop those to destroy the harm of the enzyme of collagen protein.Anthocyanidin not only helps collagen fabric to form cross-linked structure, and can help to recover because of injured and excessively crosslinked infringement of free-radical induced.Excessive crosslinked meeting suffocates connective tissue and hardens, thereby skin is corrugated and premature aging.Anthocyanidin also protects human body to avoid the sunlight injury, promotes to cure psoriasis and senile plagues.Anthocyanidin also is the fabulous additive of the skin cream of local application.

(5) cholesterol

Cholesterol is the fundamental component of cell membrane, for producing hormone and promoting transporting of fatty acid to say that it has played pivotal role.But too much cholesterol is a potential bad sign.Anthocyanidin and ascorbic combination can make cholesterol decomposition, become bile salts, and then get rid of external.Anthocyanidin has been accelerated the decomposition and the eliminating of deleterious cholesterol.Here, the conspiracy relation between vitamin C and the anthocyanidin is confirmed again.

(6) heart protective agent

Anthocyanidin not only helps to recover skin elasticity, and helps joint, tremulous pulse and other tissue (like heart) to keep normal function.Vascular system is responsible for blood flow, delivers to all cells and tissue to blood.Therefore, no matter be that anthocyanidin all is that find so far, best heart protective agent from long-term and short run effect.Anthocyanidin also suppresses histamine and produces, thereby can reduce inflammation, and helps tremulous pulse to resist the impact that those cause cardiovascular diseases's mutagen.

Although the Frenchman eats food rich in fat usually far more than the American, it is a lot of less that the Frenchman dies from cardiopathic ratio American, and the Frenchman is famous to like to drink, and normally has meal and must drink.The number that France drink gentleman suffers from coronary heart disease is also low by 30% to 40% than the crowd who is not stained with wine, and this is because contain anthocyanidin in the Bordeaux Rouge.

(7) allergy and inflammation

Anthocyanidin not only helps to slow down the cardiovascular inflammation, and also helps the treatment numerous disease, for example, and allergy, asthma, bronchitis, Hay Fever, rheumatoid arteritis, injured, the pressure ulcers of moving etc.Will discharge a kind of chemical compound that is named as histamine during the human body inflammation, it can bring out all symptoms of above-mentioned disease.Anthocyanidin suppresses to produce the enzyme that histamine needs, and prevents to generate histamine, thus dephlogistication.Anthocyanidin is the famous inhibitor of histamine, but it does not hinder other enzyme.Now, our health has too much born chemical substance and the pollutant from food, drinking water, air and animals and plants (like pollen), and its result is just allergy has become very general disease.About the ability of anthocyanidin antiallergic and anti-inflammatory, many documents are all on the books.The athlete appreciates anthocyanidin very much because anthocyanidin make the joint flexibly, can repair the collagen protein in the connective tissue, and alleviate edema.Also have report, anthocyanidin improves many people's arthritis.

(8) varicosis

Cirsoid disease can comprise pain, scratches where it itches, burns, fatigue.Serious varicosis can cause heart disease, apoplexy, thrombophlebitis, pulmonary infarction etc.Doctor Haake did clinical research in the Hamburg, Germany, found that anthocyanidin is of value to the varicosis patient.The patient who participates in test has 110 people, and wherein 41 people have shank spasm disease.Patient's every day oral 90 milligrams of anthocyanidin, 77% experimenter has major improvement.In addition, 93% shank spasm patient's disease disappears.

(9) brain function

Anthocyanidin can help to promote memory, the risk of delaying senility and apoplexy.Even after apoplexy took place, anthocyanidin can help improve memory and brain function, and this fact is proved by clinical research.For example, for child's hyperfunctioning sick (or hyperkinetic syndrome), often be to cure with the Western medicine that is called Ritilan.This Western medicine has the side effect that hinders child's growth, and the patient possibly produce dependency to it, in case cut out, the patient will produce appetite to be increased, diseases such as depression and lethargy.Some the report recommends, substitute Ritilan not only effectively but also safety with the anthocyanidin natural tonic.

(10) improve anoxia

Anoxia just is meant and lacks oxygen for a long time that this has irremediable injury to health.Old people's anoxia possibly cause problem mentally and physically, as suffering from Alzheimer (A Erchamo) disease.Aged people, blood circulation is often not so good.Anthocyanidin has been removed free radical, has suppressed the destruction of capillary rupture and surrounding tissue.Anthocyanidin has also improved the blood capillary state, the cerebripetal blood circulation of enhanced flow, and therefore, brain can obtain more oxygen.

(11) premenstrual syndrome

Clinical experiment is the result show, anthocyanidin can alleviate (anxiety) syndrome before the warp that torments the women.Because the hormone disequilibrium many kinds of diseases can occur psychologically and on the health.The body fluid retention effects blood proper flow, the result, brain, ovary and uterus can not get enough oxygen.Hypoglycemia also possibly be a kind of factor of bringing out premenstrual syndrome.This disease possibly show as following any or whole disease: mammary swelling and tenderness, sore waist and aching in the waist and the back, abdominal part bulge, muscle spasm, personality change and (as fly into a rage; Fly into a rage, even want to commit suiside), depressed, tired, faint, insomnia, arthralgia, headache, urine retention.

In the research of a premenstrual syndrome, have 165 women's every day oral 200 milligrams anthocyanidin.After two months, 60% body of women disease has disappeared.After four months, 80% women's physical symptom has not had, and the disease of half women's psychological aspects has not existed yet.

Much nourish the body in addition about anthocyanidin, wholesome research report, it comprises following (but not limiting to) these aspects: improved liver function, reduced the risk of phlebitis and cancer, improved multiple sclerosis and prevention cataract etc.

4, the safety of anthocyanidin

Masquelier professor and many scientists utilize anthocyanidin, done the research of for many years various clinical, chemical examination, toxicity and pharmacokinetics.

Extensive experimentation has proved that anthocyanidin is tonic nontoxic, not carcinogenic, that do not have antigenicity, unlikely fetal anomaly.Professor Masquelier points out that he had four more than ten years with the anthocyanidin preventing and treating diseases, never sees direct or indirect toxicity.

5, taking dose

In the many clinical reports of big number, the adult takes the anthocyanidin between 100 and 200 milligrams every day.Expert advice, maximal dose are to calculate in body weight one milligram ratio, one pound of every day, and be one to fortnight the course of treatment.Dosage reduces then, maintains every day between 50 and 100 milligrams.The time hypersensitive, a roughly method is arranged when sick or unexpected, every day oral 150 to 300mg, go over up to crisis.Child's dosage is normally adult half the.

Two, the research overview of phosphatide complexes

People such as Italy scholar Bombardelli chance in the research liposome: the natural flavone compounds has special affinity to phospholipid; The two can combine to form complex; And show and remarkable different biological characteristics of parent drug and pharmacologically active; Discovering afterwards, the natural component and the natural extract of a lot of types can be processed phosphatide complexes, and this type phosphatide complexes is named as Phytosome.

After active skull cap components and phospholipids incorporate form complex; Usually show and remarkable different physicochemical property and the biological natures of former chemical compound; Can improve the fat-soluble and stable of water soluble drug significantly, improve its transmembrane transport performance, and then improve bioavailability of medicament; Have the release of delaying simultaneously, reduce advantages such as body internal diabetes removal rates.Therefore, phosphatide complexes is considered to improve a kind of effective means of Chinese medicine water soluble ingredient oral administration biaavailability.

Phosphatide complexes (phytosome) means in non-proton carrier series solvent, and medicine combines with certain proportion relation with phospholipid and the complex that forms, is formed through Van der Waals force or hydrogen bonded in aprotic solvent by medicine and phospholipid.

Phospholipid is the important component part of cell membrane; Its molecular structure is made up of polar head and nonpolar amphipathic afterbody; Owing to having the strong electronics that gets, oxygen atom in the hydroxyl on the phosphorus atoms in the phospholipid is inclined to; Nitrogen-atoms has strong betatopic tendency, therefore under certain condition, can with the compound generation complex of the medicine of a fixed structure.And because phospholipid molecule amphipathic, also be very effective emulsifying agent.

Phospholipid is a kind of phosphorated lipid material; Be divided into natural and synthetic phospholipid two big classes; Wherein natural phospholipid mainly contains soybean phospholipid and egg yolk lecithin; Be phospholipid commonly used in the pharmaceutical preparation, it consists of phosphatidylcholine and (is called for short: PC), PHOSPHATIDYL ETHANOLAMINE (is called for short: PE), phosphatidylinositols (is called for short: PI), serinephosphatide (is called for short: PS) with phosphatidic acid (abbreviation: PA) etc.Be mainly as pharmaceutic adjuvant and contain the more natural phospholipid of PC.Synthetic phospholipid is product-free still at home, abroad mainly is that dipalmitoyl phosphatidyl choline (is called for short: DPPC) use more, but very expensive.

Phytosome is similar with Liposome aspect a lot; Also be made up of phospholipid although difference is liposome, its volume ratio phosphatide complexes is much bigger, and the more important thing is; In liposome; Phospholipid reality does not combine with the drug molecule that is wrapped, and the phosphatide complexes new molecule that to be phospholipid combine with natural component produces, significant change has taken place in its biological characteristics.

The medicine of many determined curative effects since the drawbacks limit that self property exists their application.Like some medicine stronger untoward reaction is arranged, what have has bigger zest to tissue and organ, and in gastrointestinal tract, being difficult for of having is absorbed etc.Recent study result shows that some medicine can form complex with phospholipid, and the complex of medicine and phospholipid not only can strengthen the lipotropy of medicine, reduces the untoward reaction of medicine, and can strengthen pharmaceutically active, improves drug bioavailability.

(1) phosphatide complexes

Phosphatide complexes (Phytosome) is comparatively stable chemical compound or the complex that medicine and phospholipid molecule form through the electric charge migration.It can change the physicochemical property of parent drug, improve bioavailability, and method for preparing is simple, and is with low cost.Aspect pharmaceutical preparation, use in recent years and increase gradually, particularly obtaining bigger achievement aspect the research of Chinese medicine phosphatide complexes.

1, the formation mechanism and the fundamental characteristics of phosphatide complexes

In the structure of phospholipid on the phosphorus atoms oxygen atom in the hydroxyl stronger electronics tendency is arranged, and nitrogen-atoms has stronger betatopic tendency, therefore under certain condition, it can generate complex with the drug molecule of a fixed structure.As in the structure of breviscapine, oxygen and the oxygen on the phenolic hydroxyl group on the carboxyl all have elecrtonegativity, all can produce dipole one dipole active force with the quaternary amine nitrogen of positively charged property in the lecithin and form complex; Puerarin and lecithin combine to form phosphatide complexes J through the Van der Waals force between polar fraction.After medicine and phospholipid formed complex, physicochemical property, BA etc. all can take place to change significantly, showed a lot of and parent medicine different characteristic.The change of physicochemical property such as fat-soluble obvious enhancing, significant change etc. also can take place in fusing point, absorptance, spectral signature etc.The change of BA as the activity of phosphatide complexes than parent drug is stronger, bioavailability is higher, toxic and side effects is littler.

2, the preparation of phosphatide complexes

About the existing a lot of research reports of the preparation of phosphatide complexes; Usual way is that medicine and phospholipid are placed aprotonic solvent such as aromatic hydrocarbons, halogen derivatives or some cyclic ethers (like oxolane, chloroform, methanol, ether, dioxane etc.), handles making through means such as heating, stirring, backflows.The complex for preparing can or go down to desolventize in vacuum and obtains through evaporation, and the also available freeze-drying or the non-solvent sedimentation method are separated and obtained.The factors such as rate of charge of the selection of reaction dissolvent, response time, reaction temperature, concentration of reactants, medicine and phospholipid all can influence the combination rate of medicine, can select best preparation prescription and technology through testing sieve.

3, the discriminating of phosphatide complexes

General adopt method discriminatings such as thin layer, ultraviolet, infrared, mass spectrum and heat analysiss.Proton nmr spectra such as broad-mouthed receptacle for holding liquid celebrating state and TLC are analyzed the product that puerarin and phospholipid react, and reach a conclusion: product is complex rather than chemical compound.Equally puerarin and the product that phospholipid reacts are analyzed with ultraviolet method and wish that industry light etc. is infrared, infrared spectrum analysis shows that the spectrum of phosphatide complexes is the different spectrographic adductions with phospholipid of puerarin; Visible phosphatide complexes of ultraviolet spectral analysis and puerarin absworption peak are all in the 251nm wavelength.Reach a conclusion thus, what puerarin and phospholipid formed is phosphatide complexes.The Tang Xiao buckwheat carries out differential calorimetric scanning to breviscapine, lecithin and the complex of the two and physical mixture, and the result obviously reduces for the phase transition temperature of complex, and the phase transition temperature of physical mixture occurs overlapping, but the scope of phase transition temperature does not have significant change.

4, the physicochemical property research of phosphatide complexes

Compare with parent drug, great changes have all taken place in the dissolubility of phosphatide complexes, fusing point, absorptance, spectral signature.

5, the materia medica of phosphatide complexes research

Lecithin is the basic composition material of cell membrane; Strong with the affinity of cell membrane; Behind medicine and lecithin formation phosphatide complexes, lecithin can promote thing to get into cell from the enterocyte film that hydrophilic environment shifts through the lipophilic environment better as pharmaceutical carrier, arrives blood at last.Chemical analysis experiment through precision shows, the flavonoid phosphatide complexes of a part normally connects at least the lecithin of a part by the flavone of a part and forms.The conjugate of this fat-solubility is very easy to the cell membrane through enterocyte.

Phospholipid can also with the relevant group effect with zest and untoward reaction, or the long carbochain in the phospholipid surrounds these groups, makes the zest of medicine diminish, toxic and side effects reduces.

Phosphatide complexes can improve the fat-soluble of medicine significantly as a kind of novel medicament preparation, strengthens drug absorption, improves drug bioavailability, reduces adverse effect etc.This all has important theory and practical significance for exploring China's natural drug, quickening Study on Modernization progress, raising Chinese medicine industrial technology level and the clinical practice quality of Chinese medicine compound preparation.But aspect a lot, especially aspects such as the preparation technology of complex, analysis discrimination method and mechanism of absorption are still waiting further research and exploration.

(2) progress of active ingredient of Chinese herbs phosphatide complexes

After active ingredient of Chinese herbs and phospholipid formed complex, physicochemical property had certain change than active ingredient of Chinese herbs, thereby drug treating time is prolonged, and untoward reaction reduces, and pharmacological action strengthens, and bioavailability improves.Conclusion active ingredient of Chinese herbs phosphatide complexes has excellent application value and DEVELOPMENT PROSPECT.

Though a lot of active ingredient of Chinese herbs have definite pharmacodynamics effect; But because of physicochemical property such as the characteristics such as polarity is big, fat-soluble difference of itself; Cause orally difficult absorb, bioavailability is low, or limited its application clinically because of it is big to gastrointestinal irritation property, untoward reaction is more.In recent years, countries in the world document (Giori, Andrea.Phospholipid complex ofcurcumin having improved bioavailability [P] .United States, 20090131373,2009-03-21.; Y Li, W S Pan, S L Chen, et al.Studies on preparation of puerarin phytosomes and their solid dispersions [J] .Chin Pharm J, 2006,37 (5): 695.; Maiti; K.Mukherjee; A.Gantait; Et al.Enhanced therapeutic benefit of quercetin-phospholipid complex in carbon tetrachloride induced acute liver injury in rats:a comparative study [J] .Iran J.Pharmacol Ther, 2005,4 (2): 84.; Zhai Guangxi; Lou Hongxiang; Zou Lijia, etc. the progress of medicine phosphatide complexes [J]. Chinese Pharmaceutical Journal, 2001; 36 (12): the active ingredient of Chinese herbs of 800.) having reported a lot of types all can form complex with phospholipids incorporate, and shows and parent drug different biological characteristics and pharmacologically active.Phosphatide complexes (phospholipid complex or phytosome) is a kind of novel drug-loading system that Italian scholar Bombardelli finds when the research liposome; After active ingredient of Chinese herbs and phospholipid form phosphatide complexes under certain condition, the medicine lipotropy is obviously strengthened, prolong action time; Untoward reaction reduces; Simultaneously can strengthen the absorption in gastrointestinal tract, improve bioavailability, strengthen pharmacological action.

1, the preparation of active ingredient of Chinese herbs phosphatide complexes and discriminating

The preparation of active ingredient of Chinese herbs phosphatide complexes generally in aprotonic solvent (left high sea, Shang Jingchuan. the progress of Chinese medicine phosphatide complexes [J]. China Dispensary, 2007; 18 (27): 2149.) react; Like aromatic hydrocarbons, halogen derivatives or some cyclic ethers etc., this is because phospholipid and the bonded key of medicine are intermolecular force, in the ionized solvent of ability; This active force is easy to broken, so adopt aprotic solvent.Reaction can be handled through means such as heating, stirring, backflows and make; The complex for preparing can or go down to desolventize in vacuum and obtains through evaporation; Also available freeze-drying or the non-solvent sedimentation method (Ying Li; Da Jian Yang, Shi Lin Chen, et al.Comparative physicochemical characterization of phospholipids complex of puerarin formulated by conventional and supercritical methods [J] .Pharm Res; 2008,25 (3): 563.) separation obtains.Preparation technology's evaluation index of phosphatide complexes generally adopts the percentage rate that combines of medicine and phospholipid; Factors such as the rate of charge of the selection of the composition of phospholipid and quality, reaction dissolvent, medicine and phospholipid, response time, reaction temperature, reactant concentration all can influence the combination rate of medicine, can select best preparation technology through testing sieve, as: (Wu Jianmei such as Wu Jianmei; Chen Dawei; Liu Yanli. baicalin phosphatide complexes preparation technology's research [J]. CHINA JOURNAL OF CHINESE MATERIA MEDICA, 2001,26 (3): 166.) percentage rate that combines with baicalin and phospholipid is an evaluation criteria; Adopt single factor investigation and orthonormal design of experiments to carry out systematic research; Having confirmed the optimum preparating condition of baicalin phosphatide complexes, at room temperature is reaction dissolvent with the oxolane promptly, and the response time is 1.5h.(be called for short: PC) phospholipid of content is different with the optimum response ratio of baicalin, and the formation that draws the baicalin phosphatide complexes receives the influence of solvent, reactant concentration and reactant ingredient proportion bigger for different phosphatidylcholines.

The discriminating of Chinese medicine phosphatide complexes generally can pass through nuclear magnetic resonance, NMR (be called for short: NMR), infrared (be called for short: IR), ultraviolet (be called for short: UV) wait with heat analysis (Sun Yan, Gao Er, Wang Ruqin. active ingredient of Chinese herbs phosphatide complexes progress [J]. medical science is summarized; 2007,13 (11): 875.) data of aspect are judged the formation of complex, as week waiting (Zhou Jing quietly; Wu Zhenghong, flat its ability, etc. the preparation of indirubin phosphatide compound and the research of physicochemical property [J]. Chinese Journal of Pharmaceuticals; 2006; 37 (6): 394.) through nuclear magnetic resonance spectroscopy phospholipid and indirubin phosphatide compound, the result shows that the position at phospholipid and indirubin phosphatide compound wave spectrum peak is almost constant, the latter's wave spectrum peak slightly broadens; This shows that the phosphorus atoms in the phospholipid molecule does not change, and does not produce new chemical bond; (Song YM, Zhuang J, Guo JX such as Song; Et al.Preparation and properties of a silybin-phospholipid complex [J] .Pharmazie; 2008,63 (1): 35.) adopt ultraviolet spectroscopy that silibinin and its phosphatide complexes are analyzed, the result shows; The ultraviolet spectra of silibinin and its phosphatide complexes is as broad as long; This shows that medicine and phospholipids incorporate do not influence the conjugated system of silibinin, and its chromophore does not change yet, and therefore we can say that bright silibinin phosphatide complexes is not new chemical compound.

2, the physicochemical property of active ingredient of Chinese herbs phosphatide complexes

2.1 the diffraction analysis x-ray diffraction pattern of the X-ray of active ingredient of Chinese herbs phosphatide complexes shows that mostly active ingredient of Chinese herbs is crystal type; And demonstrating unformed characteristic behind the formation complex, this possibly be because active ingredient of Chinese herbs combines with the polar end of phospholipid is directed, thereby makes medicine be in the state of high degree of dispersion; The crystal characteristic of himself is suppressed; As Liang Na etc. (Liang Na, Shi Kai, Wang Yongsheng. the preparation of ferulic acid-phosphatide complexes and physical characteristic thereof [J]. Shenyang Pharmaceutical University's journal; 2008,25 (3): 163.) respectively ferulic acid, phospholipid, ferulic acid mixture of phospholipids, ferulic acid-phosphatide complexes are carried out the X-ray diffraction analysis.The result shows that ferulic acid presents many peak crystallization on collection of illustrative plates, show that ferulic acid mainly exists with crystal type.Phospholipid then shows unformed characteristic, is merely a broadband on the diffraction pattern.In the diffracting spectrum of ferulic acid and phospholipid physical mixture, the crystalline diffraction maximum of ferulic acid still appears.In the diffracting spectrum of ferulic acid-phosphatide complexes, when the two combined with 1: 2 molar ratio, the crystal diffraction peak almost completely disappeared, and complex shows the unformed characteristic of phospholipid fully.

Show that the active ingredient of Chinese herbs phosphatide complexes compares with active ingredient of Chinese herbs 2.2 active ingredient of Chinese herbs phosphatide complexes differential calorimetric scanning method is analyzed differential calorimetric scanning method, phase transition temperature decreases, this possibly be since active ingredient of Chinese herbs with after the polar end of phospholipid combines; The hydrocarbon chain of phospholipid is free to rotate; Wrapped up the polarity part of phospholipid, the cause that makes the interchain order reduction of phospholipid aliphatic hydrocarbon is like (Song YM such as Song; Zhuang J; Guo JX, et al.Preparation and properties of a silybin-phospholipid complex [J] .Pharmazie, 2008; 63 (1): 35.) measure silibinin, phospholipid, silibinin phosphatide complexes respectively; Can find out that from the DSC curve of silibinin phosphatide complexes medicine peak and phospholipid peak all disappear, and the phase transition temperature of silibinin phosphatide complexes is lower than phospholipid.

2.3 the dispersing morphology active ingredient of Chinese herbs phosphatide complexes of active ingredient of Chinese herbs phosphatide complexes in water can form the micelle structure in water; Can be dispersed in the water; Intermolecular ordered arrangement forms the multilamellar capsule of outer appearnce like liposome; As Zhang Qian etc. (open pretty. the anti-capsular pharmacokinetic studies of hepatitis kurarinone phosphatide complexes [D]. Sichuan: Chengdu University of Traditional Chinese Medicine, 2006.) under transmission electron microscope, observe, the kurarinone phosphatide complexes disperses to be in water and the similar micelle shape of liposome.But because the generting machanism of itself and liposome is completely different, its internal structure certainly exists difference: the liposome Chinese medicine is encapsulated in the hydrophilic group interlayer of vesicle or is scattered in the interlayer of hydrophobic group of vesicle; Phosphatide complexes then is an integral body, and medicine interacts through the polar group with phospholipid and is fixed.

2.4 after the solubility property active ingredient of Chinese herbs of active ingredient of Chinese herbs phosphatide complexes and phospholipid form complex, can improve the lipotropy and the hydrophilic of medicine to a certain extent, so moisten (Xu Runchun such as spring; Lin Yanjun; Wu Pinjiang, etc. the research [J] of baicalin phosphatide complexes physicochemical property. Chinese patent medicine, 2008; 30 (6): 932.) measure the solubility property of baicalin phosphatide complexes in water and n-octyl alcohol, its meltage in water is 4.56 times of baicalin as a result; In n-octyl alcohol; Its meltage is 70.17 times of baicalin approximately, so it had both improved the hydrophilic of baicalin, has improved lipotropy again; And it is bigger that lipotropy improves degree, and physical mixture is only improving the solubility property of baicalin in water and n-octyl alcohol on the limited degree.

3, the study of pharmacy of active ingredient of Chinese herbs phosphatide complexes

3.1 pharmacodynamic study silibinin phosphatide complexes is a research phosphatide complexes early; Countries in the world are to more (the DiSario A of its research; Bendia E; Taffetani S.Hepatoprotective and antifibrotic effect of a new silybin-phosphatidylcholine-vitamin E complex in rats [J] .Dig Liver Dis, 2005,11 (37): 869.; Kidd P; Head K.A review of the bioavailability and clinical efficacy of milk thistle phytosome:a silybin-phosphatidylcholine complex (Siliphos) [J] .Altern Med Rev; 2005; 10 (3): 193.); (Chen ZY such as Chen; Xu JW, Zhang GJ.Protective reactione of Silybin-phosphate-dylcholine compound on cerebral ischemic reperfusion injury in rats [J] .Chin J Clin Rehabil, 2004; 8 (22): the rat oral gavage that 4629.) damages to Cerebral Ischemia with silibinin and phosphatidylcholine complex thereof respectively, paired observation silibinin and silibinin phosphatidylcholine complex are to the influence of rat cerebral ischemia/reperfusion injury model brain water content, pathological change, plasma malonaldehyde content.The result shows that silibinin and silibinin phosphatidylcholine complex all have protective effect to rat cerebral ischemia/reperfusion injury, and the effect of silibinin phosphatidylcholine complex is superior to silibinin.Possibly be because phospholipid can strengthen silibinin and absorb at gastrointestinal on the one hand; Thereby the bioavailability that has improved silibinin strengthens its drug effect; Phosphatidylcholine also has free radical resisting, lipoid peroxidization resistant on the other hand, possibly produce synergism with silibinin.

(Liu Jinjin such as Liu Jinjin; Wang Fang, Chen Jinhe, etc. the pharmacodynamic study [J] of Folium Ginkgo extract phosphatidylcholine complex. medical science new knowledge magazine; 2003; 13 (1): 27.) adopt Cavia porcellus working heart ischemical reperfusion injury model, (be called for short: Gb-PC) to the influence of ischemical reperfusion injury cardiac muscle cardiac function and biochemical indicator, the result shows that Gb-PC can obviously improve the systolic and diastolic function of ischemical reperfusion injury cardiac muscle to observe Folium Ginkgo extract phosphatidylcholine complex; The activity of creatine phosphokinase and lactic acid dehydrogenase in the dose dependent ground reduction arteria coronaria effluent; Reduce MDA content in the ischemic myocardial tissue, explain that Gb-PC has protective effect to the ischemical reperfusion injury cardiac muscle, its mechanism is relevant with its anti peroxidation of lipid.

Tang Xiao buckwheat etc. (Tang Xiaoqiao. the preparation of breviscapine-phosphotide compound and Pharmacological action study thereof [D]. Wuhan: the Central China University of Science and Technology; 2004.) to improve anoxia mice endurance with the breviscapine powder group of dosage better for the research breviscapine-phosphotide compound; Can better prolong that normobaric hypoxia suffocates and chemical substance due to the life span of histanoxia mice; Increase the number of times of dehiscing of the Mus head that exsomatizes, reduce the whole mice and the heart, brain oxygen consumption.

3.2 pharmacokinetic studies phospholipid is the basic composition material of cell membrane; Strong with the affinity of cell membrane, behind medicine and lecithin formation phosphatide complexes, physicochemical property can change to some extent; Thereby can improve medicine absorbs through gastrointestinal tract or percutaneous; Obtain higher blood drug level, medicine elimination in vivo simultaneously is slack-off, has significantly improved bioavailability.(Peng-FeiYue such as Yue; Wen-Jin Zhang, Hai-Long Yuan, et al.Process optimization; Characterization and pharmacokinetic evaluation in Rats ofursodeoxycholic acid-phospholipid complex [J] .AAPS Pharm Sci Tech; 2008,9 (1): 322.) studied the ursodesoxycholic acid tablet and (be called for short: UDCATablet) (be called for short:, measure the content of UDCA in the rat serum with HPLC UDCA-PLC) in the Wistar its pharmacokinetics in rats with the Ursodeoxycholic acid phospholipid complex; Divide two groups (6 at random with rat; Male) irritate clothes ursodesoxycholic acid tablet respectively and Ursodeoxycholic acid phospholipid complex (dosage is ursodesoxycholic acid 20mg/kg) result shows that the Tmax of UDCA among the UDCA-PLC is 1.6h, Cmax is 0.1346 μ g/ml, AUC is 11.437 μ gh/ml; Compare with the ursodesoxycholic acid tablet, the relative bioavailability of UDCA has improved 241% among the UDCA-PLC.

(Y.Li, W.S.Pan, S.L.Chen such as Li; Et al.Pharmacokinetic; Tissue distribution, and excretion of puerarin and puerarin-phospholipid complex in rats [J] .Drug Dev Ind Pharm, 2006; 32 (5): 413.) puerarin pharmacokinetics in Beagle dog body in research puerarin and the phosphatide complexes thereof; Adopt the HPLC method to measure puerarin content in the dog blood, intersect oral puerarin and phosphatide complexes thereof with dog (3, male) on the same group; The result shows that medicine-time process of puerarin meets opening two chambers first order kinetics model; The AUC of oral puerarin, Cmax, Tmax are respectively (10.91 ± 4.83) mgh/L, (3.00 ± 1.13) mg/L, (1.62 ± 0.30) h, and the AUC of puerarin phosphate composition, Cmax, Tmax are respectively (13.67 ± 2.72) mgh/L, (1.91 ± 0.51) mg/L, (2.38 ± 1.27) h, and statistical result shows that significant difference is arranged between the AUC; Can improve puerarin in the intravital absorption of Beagle dog it is thus clear that form phosphatide complexes, increase its bioavailability.

(Xiao YY, Song YM, Chen ZP such as Xiao; Et al.The preparation of silybin-phospholipid complex and the study on its pharmacokinetics in rats [J] .Int J Pharm, 2006,307 (1): 77.) studied silibinin phosphatide complexes and silibinin-N-cardiografin in its pharmacokinetics in rats; After the result showed rats gavaged silibinin phosphatide complexes and silibinin-N-cardiografin, medicine-time process of silibinin met open single chamber first order kinetics model, the AUC of silibinin phosphatide complexes; Cmax, Tmax is respectively 1020.33ngh/ml, 126.72ng/ml; 10min, and the AUC of silibinin-N-cardiografin, Cmax; Tmax is respectively 235.81ngh/ml, 104.29ng/ml, 5min; This shows that the formation of phosphatide complexes has increased the lipotropy of medicine, thereby its bioavailability is improved.

4, the application of active ingredient of Chinese herbs phosphatide complexes

4.1 functional cosmetics skin is the maximum organ of human body, is made up of epidermis, corium and subcutaneous tissue, can protect body to avoid the infringement of physical property, mechanicalness, chemical and pathogenic microbes.Dermatosis and delaying sanilities such as xerosis cutis, eczema, acne, acne can cured or prevent to some active ingredient of Chinese herbs; Have effects such as antiinflammatory, detumescence, analgesia, anti-fungal infection like the flavonoid active component; Not only have medical value to also have beauty functions, thereby but because the Transdermal absorption ability is used restricted.Phospholipid and active ingredient of Chinese herbs form behind the complex as functional cosmetics; It is externally applied product (the Giorgio P that the effect of existing cosmetics also has medicinal efficacy simultaneously; Pirgiorgio A.Resveratrol-phospholipids complexes, their preparation, and pharmaceutical and cosmetic composition containing them [P] .United States; 20040116386,2004-06-17.).The polymeric phosphatide complexes of Semen Ginkgo procyanidin (Gupta Amit; M.S.Ashawat; Saraf Shailendra; Et al.Phytosome:A Novel Approach Towards Functional Cosmetics [J] .J Plant Sci, 2007,2 (6): 644.) be applied to 18 health volunteer's tests (they all scytitis and erythema have occurred because of ultraviolet radiation); Experiment shows that phosphatide complexes has the effect of protectiving ultraviolet injury preferably, can be used for skin antiinflammatory and microcirculation improvement; Terpenoid in the Herba Centellae extract and phospholipid can help wound healing after forming complex, cure skin injury.The phosphatide complexes of functional cosmetics has high similarity because of itself and human skin lipid; So have the good skin permeability; Thereby improved the absorption of active component, required dosage reduces, and prepares used phospholipid in addition except that making carrier; Also to the skin Yellow Gentian Extract, and biological degradability in the good body is arranged.

4.2 medicine active ingredient of Chinese herbs phosphatide complexes at first is used for cosmetics, has expanded to the pharmaceutical preparation industry at present.Active ingredient of Chinese herbs and phospholipid form complex, and its character also changes, and has following advantage: 1. can significantly improve drug bioavailability, the fast and interior absorption enhancing of intestinal of travelling speed.2. after the absorption of main active improved, the corresponding reduction of required dosage still can reach good therapeutic effect.3. the phospholipid that is used to prepare phosphatide complexes not only can be used as carrier, also has hepatoprotective effect.4. as far as hepatic,, promoted hepatic targeting through strengthening the dissolubility of medicine in bile.Common industrialized phospholipid complex formulation is seen Ajay Semalty at present; Mona Semalty; M.S.M.Rawat.The Phyto-phospholipid complex phytosomes:A potential therapeutic approach for herbal hepatoprotective drug delivery [J] .Phcog Rev; 2007,2 (1): 369.; Gupta A, Saraf S.Phytosome:A novel approach for phytoconstituents [J] .Plant archives, 2007,7 (2): 457..

4.3 nutrient and healthcare products lecithin is described as and protein, vitamin " the 3rd nutrient " arranged side by side; Even as appetizing food; And itself and some active ingredient of Chinese herbs with health role; As: the epicatechin acid esters in the green tea extract, the Oligomeric Proanthocyanidins in the Semen Vitis viniferae extract, the flavone in the Fructus Crataegi extract, (the Gupta A such as ginsenoside in the Radix Ginseng extract; Saraf S.Phytosome:A novel approach for phytoconstituents [J] .Plant archives, 2007,7 (2): 457.; Kidd, P.M., 2007.Phytosome, highly bioavailable plant extracts.www.indena.com.) form phosphatide complexes after, can be absorbed better, as nutrient and healthcare products, make consumers in general's income.

5, prospect

At present, the research of Chinese medicine preparation is also more and more, but compares with Western medicine prepn; The traditional Chinese medicines preparation still exists technology to fall behind, and dosage form is outmoded, and quality is low; The preparation basic research is weak; Therefore problems such as novel form research and development ability difference develop the Chinese medicine novel form, improve the bioavailability of active ingredient of Chinese herbs and the focus that the pharmacology curative effect has become countries in the world field of medicaments research.The active ingredient of Chinese herbs phosphatide complexes is as a kind of new potential drug-supplying system; Have a good application prospect undoubtedly; It not only has some characteristics of liposome; As: reduce toxicity, delay release, target administration, minimizing dosage etc.; Also,, therefore be regarded as a new breakthrough in traditional medicine field so phosphatide complexes has more excellent characteristics than liposome because the drug molecule phosphatide complexes of equivalent is more much smaller than liposome volume; The active ingredient of Chinese herbs phosphatide complexes be formed with the foundation that modern pharmacy possibly is provided for pharmaceutics of Chinese drugs; But a lot of aspects are such as the evaluation index of active ingredient of Chinese herbs phosphatide complexes, analyze discrimination method and mechanism of absorption etc. and await further research and explore (changeing the http://www.studa.net in Chinese paper download center, the author: Ge Yanru, Zhang Chunyan, Ji Shunli).

Three, the research overview of vascular dementia

Dull-witted (Dementia) is the acquired and persistence disturbance of intelligence syndrome that produces owing to disordered brain function, comprises that in various degree memory, language, visual space function, abnormality of personality and cognitive competence reduce.Vascular dementia is caused by cerebrovascular, is mainly caused by cerebral hypoxia ischemia.

Dull-witted main type is to comprise vascular dementia (vascular dementia; Be called for short: VD), the senile dementia in the senile dementia (Alzheimer ' s Disease; Or the title alzheimer's disease, or claim to be called for short Alzheimer, Alzheimer's disease: AD) etc.

Senile dementia comprises senile dementia, multiple infraction type dementia (Multimfarct Dementia), alcoholic dementia disease (Alcoholic Dementia) and normal brain activity setting-out brain disease (Normal Pressure Hychocephalus), and wherein senile dementia is the main type in the senile dementia.Because AD falls ill in old people more than 60 years old well, so custom is called as senile dementia or degenerative brain disorder.

The common clinically multi-infarct dementia of vascular dementia, large tracts of land cerebral infarction dementia, thalamic dementia and types such as concurrent dementia of hemorrhagic cerebrovaseular and Combination dementia.Clinical manifestation is depression, delirium, aphasia etc.Pathological change is that multiple lacuna sexually transmitted disease (STD) becomes or large tracts of land infarction kitchen range and atherosclerotic lesion (referring to the neurological, the 4th edition, People's Health Publisher).Clinical treatment can use the cholinergic inhibitor, and blood vessel dilating, cerebral blood flow increasing amount perhaps utilize cerebral protective agent-antioxidant or free radical scavenger can improve ischemia, the caused pathology damage minimizing brain cell necrosis of anoxia and apoptosis, protection cerebral tissue.

The vascular dementia person only shows nearly dysmnesia in early days, and memory far away keeps better relatively; With the increase of the course of disease, memory ability far away is lost gradually, and hypophrenia is gradual and increases the weight of.

And alzheimer disease just shows far and near dysmnesia in early days; Hypophrenia demonstrates a kind of (Guo Mingying of progression slowly; Korea S's tinkling of pieces of jade, etc. the comparative study of alzheimer disease and vascular dementia disturbance of intelligence. the journal .2007 of Qinghai Medicine College, 28 (2): 125-127.).

The vascular risk factor is all being played the part of important role in the morbidity of vascular dementia and Alzheimer, so there is theory to think that Alzheimer and vascular dementia possibly not be two independent illness.The general character research of Alzheimer and vascular dementia is carried out in various aspects such as risk factor, pathogenesis, Pathophysiology, iconography changes widely.

Vascular dementia is the carrying out property decline property disease by the acquired Premium Features of cerebral cortex due to apoplexy repeatedly or the long-term chronic cerebral ischemia.

Show that in America and Europe investigation dementia incidence rate is about 1.1% among the old man more than 55 years old, wherein the Alzheimer disease accounts for top priority (incidence rate about 7.7 ‰); Be about 50%～60% of dull-witted total number of persons, and VD occupies time (incidence rate about 1.5 ‰), accounts for 10%～20%; And with age, the VD proportion significantly increases (1. Ott A, Breteler MMB; Harskamp F, Stijnen T, Hofman A.Incidence and risk of dementia:the Rotterdam study.Am.J.Epidemiol.; 1998,147 (6): 574 – 580.; 2. Di Carlo A, Baldereschi M, Amaducci L, et al.Incidence of dementia, Alzheimer'sdisease, and vascular dementia in Italy.The ILSA Study.J.Am.Geriatr.Soc., 2002; 50:41-48.).

The nearest research of China shows that also the AD incidence rate is about 3.5% among the old man of over-65s, VD incidence rate 1.1% (ZhangZX; Zahner GE; Roman GC, et al.Dementia subtypes in China:prevalence in Beijing, Xian; Shanghai, and Chengdu.Arch Neurol.2005; 62 (3): 447-453.).

Owing to also lack the cognitive impairment pattern that is applicable to diagnosis VD at present; Discriminating and " mixed type " dull-witted (AD+ cerebrovascular for the diagnosis of AD and VD; Be called for short: diagnosis AD+CVD) still has certain challenge (1. plain chrysanthemum of fourth; Li Yunxia. the clinical diagnosis of vascular dementia and treatment. foreign medical science: cerebrovascular disease fascicle .2005,13 (9): 676-680; 2. Erkinjuntti T:Vascular dementia:challenge of clinical diagnosis.Int Psychogeriatr.1997; 9:51 – 58.); So someone thinks that vascular dementia possibly be a dull-witted type the most common among the old people; Mainly be because a lot " mixed types " dull-witted (AD+CVD) possibly be diagnosed as AD (1. Rom á n GC.Vascular dementia may be the most common form ofdementia in the elderly.J Neurol Sci.2002,203-204:7-10; 2. the plain chrysanthemum of fourth. vascular dementia. Chinese gerontology magazine .2003; 23 (4): 200-202.).

The incidence rate of VD is at men and women's no significant difference (Anderson K; Launer LJ; Dewey ME, et al.Gender differences in the incidence of AD and vascular dementia:the EURODEM studies.Neurology.1999; 53:1992 – 1997.); Its most significant risk factor is age [HebertR., Lindsay J., VerreaultR.; Rockwood K.; Hill G., Dubois MF.Vascular dementia:incidence and risk factors in the Canadian study of health and aging.Stroke, 2000; 31 (7): 1487-1493.], according to Epidemiological study, in greater than 60 years old crowd, the sickness rate of 5 years old VD of every increase of age just increases by 1 times.

Two main causes that cause VD are that (ischemic heart disease, be called for short: IHD), and the two all is a commonly encountered diseases in the old people for apoplexy and ischemic heart desease.Only with regard to apoplexy, the present annual whole world increases apoplexy patient 1,000 ten thousand newly, wherein China (the 1. Feigin VL that just has 2,000,000; Lawes CMM, Bennett DA, et al.Stroke epidemiology:a review of population-based studies of incidence; Prevalence; And case-fatality in the late 20th century.LancetNeurol., 2003,2 (1): 43-53; 2. Wu Zhao revives, Yao Chonghua, Zhao's winter. the epidemiological study of Chinese population stroke onset rate, mortality rate. and Chinese epidemiology magazine, 2003; 24 (3): 236-239.), sickness rate is high especially in the old people, thereby causes the concurrent VD of a lot of patients.The national dull-witted epidemiology statistics data of China also shows the aging along with population, the change of dietary structure, and the morbidity of VD is and increases progressively trend.

Along with the aging of population, the sickness rate of cardiovascular and cerebrovascular disease raises year by year, and concurrent VD patient also increases gradually.VD is the grievous injury patient health not only, brings long-term misery to the patient, influences patient's quality of life, brings white elephant also for society and family, has caused the common concern of countries in the world, and it is an important topic in the geriatrics field.Generally believe that the age influences through many-side the effect of VD; For example the aging change of metabolism, blood brain barrier and the autonomic function aspect of self the regulating of brain, cell makes cerebrovascular be easy to suffer damage, and encephaloclastic accumulative total effect also is one of reason.

The diagnosis of vascular dementia must meet three conditions: 1. dull-witted; 2. cerebrovascular; 3. the two is closely related more than.Dull-witted and apoplexy is maintained close ties with in time, takes place dull-witted usually after the apoplexy in 3 months.Cardinal symptom is: 1. the cerebral tissue pathomorphism changes; 2. hypomnesis.

About the treatment of VD, at present still do not have sure Therapeutic Method and can change the whole course of disease, because downright bad brain cell can not reverse behind the cerebral infarction, mainly be to alleviate stage of attack symptom, prevention infringement etc. again to the treatment of the brain cell of blood supply insufficiency.Normal employing actively improves the method for brain cell oxygen supply, microcirculation improvement, the new thrombosis of prevention and re-infarction etc.Improve cerebral circulation treatment commonly used 9,10-Dihydroergotoxine class, calcium ion antagonist, nicotinic acid class, other medicament for expanding vascellum and medicament for resisting platelet aggregation arranged.Cholinesterase inhibitor is a developed recently dementia treatment medicine comparatively rapidly, example hydrochloric acid donepezil and rivastigmine-hydrogentartrate.Also often should require mental skill clinically and protect medicine such as calcium ion antagonist (nimodipine and the spirit of western pyrrole); Free radical scavenger (vitamin E, vitamin C and gingko leaf preparation) etc.But from the present Clinical Application effect of said medicine, curative effect is general, and major part all only is anti symptom treatment, can not improve mortality rate.There are weak points such as active component is unclear, mechanism of action not clear, quality is unstable, toxic and side effects is big mostly in preparations such as some Chinese medicine preparation such as Folium Ginkgo, Herba Erigerontis, Radix Et Caulis Acanthopanacis Senticosi at present, are difficult to meet clinical needs.

Therefore; At present countries in the world still do not have effective ways and medicine to controlling this course of disease progress, the drug research of the prevention of VD, treatment and prognosis and exploitation become the research topic paid much attention to various countries (the 1. plain chrysanthemum of fourth. vascular dementia. Chinese gerontology magazine .2003; 23 (4): 200-202; 2. Chen Chunlei, Qiu Zhihui, Su Shixin, Zheng Zhongcheng, Chen Peifen, Ye Zhiping. the therapeutic advance of vascular dementia. the new .2007 of medical science forum of China, 7 (7): 59-62.).

Four, the progress of senile dementia

1, general introduction

Topmost disease type is AD in the senile dementia.AD is a kind of chronic former, is the old central nervous system degenerative disease basis, carrying out property cerebral degeneration with the brain aging; It is a kind of progressive neurodegenerative diseases that betides geratic period or presenium; Be a kind of common stiff reaction after the entering declining years, belong to the presenile dementia disease of improper aging state.

AD is a kind of serious, degeneration brain illness.AD is with hypomnesis, cognitive disorder, personality changes; Dystropy and linguistic competence's forfeiture are characteristic; The comprehensive obstacle and the cognitive function that mainly show as the acquired Premium Features of cerebral cortex are obviously impaired; Cerebral functional deterioration decline, i.e. memory function decay and identification ability obstacle rapidly, and with the change of behavior or personality aspect.Its Clinical symptoms is carrying out a property cognitive dysfunction, and clinical manifestation is amnesia, cognitive disorder, personality changes, and its pathological characters is that neurofibrillary tangles in the neuron, extracellular senile plaque deposition and neuronal quantity reduce.Because when people or other animal were old and feeble, many degeneration can take place body itself changed, therefore, AD is as Senile disease, and it takes place with aging substantial connection is arranged.

Along with global aged tendency of population, the sickness rate of alzheimer disease also increases thereupon.According to the Rotterdarm result of study, the annual sickness rate of old people was that 1.4%, 70～74 years old person is that 3.9%, 75～79 years old person is 16.7 ‰ in 65～69 years old, increased to 45.4 ‰ during by 85 years old.This sick genesis mechanism has various theories, but the definite cause of disease does not still have final conclusion.

At present; The whole world has more than 5,000 ten thousand old people to suffer from alzheimer disease approximately, and the trend of continuous rejuvenation is arranged, become the social problem that can not be ignored and domestic problem (Zhang Juntian. the pathogenesis of senile dementia and control medicine. medical Leader; 2002,2l (8): 469.471).And senile dementia patient's mean survival time (MST) is 5.5 years, makes this disease become main one of disease that causes death of modern society old people.

But because the cause of disease of AD is not exclusively clear and definite; Still there is not the effectively preventing medicine so far; This disease is still one of comparatively thorny disease in the whole world at present; The control of AD has become a medical treatment outstanding, that need to be resolved hurrily and social problem, and the new Therapeutic Method of active research exploitation is an instant thing.Such disease course is generally longer, is about 3～20 years.To the long-term nurse fees of AD patient is huge, and not only there is very big influence in the medical security system to entire society, also is great challenge to nuclear family's pattern of Chinese Future Society.Therefore, it takes place to give society, family all to bring heavy financial burden and mental burden, brings great misery also for patient self.

2, pathological manifestations and pathogenesis

The main pathological manifestations of AD is the formation at forebrain substrate, Hippocampus and cerebral cortex extracellular neuritis's speckle, and the appearance of neurofibrillary tangles and neurocyte and number of synapses purpose reduce in the cell.Mainly (be called for short: (Beta-Amyloid Protein is called for short: A β) assemble and form protein hydrolysate amyloid-beta APP) speckle by amyloid precursor protein.Under normal circumstances, the APP hydrolyzate is the A β of solubility ₄₀, and under pathological conditions, the APP hydrolyzate is the A β of the easy formation fiber of length _1-42, it exists with a kind of immature, non-fibrous dispersive speckle form earlier, finally passes through conformational change, aggregates into a kind of deleterious fibrous material, promptly forms speckle.Neurofibrillary tangles mainly is present in the interior and unusual neural axon of the neuronic born of the same parents of taper; It is a water-insoluble structure; It mainly consists of the Double helix fibre structure of diameter 10nm, and (be called for short: PHF), main component is a kind of microtubule bindin, i.e. Protein tau; The proteic interaction of normal Protein tau ability stabilize microtubules, the Protein tau among the PHFs then exists with the form of ultra phosphorylation.

The etiology of AD is complicated, mainly contains two kinds of hypothesis at present: Protein tau hypothesis and amyloid hypothesis.The main argument of Protein tau hypothesis is: because the ultra phosphorylation of Protein tau causes the instability of tubulin, cause that the Golgi body structure changes, thereby influence the metabolism of APP, produce excessive A β.This hypothesis also is not confirmed at cell or animal individual level.The main argument of amyloid hypothesis is: the gathering of amyloid around neuropathy takes place in the extracellular causes the formation of neuritis's speckle and the formation of neurofibrillary tangles subsequently.Therefore, amyloid is the cause of AD pathology chain formation.Support that the evidence of amyloid hypothesis is very extensive.

The senile plaque that the amyloid-beta abnormal deposition forms is the typical pathological characters of AD.To sexually revise the activation prompting A β with glial cell possibly be the reason that causes neuron loss and inflammatory reaction among the AD to significant neurological around the speckle.In nerve retrograde affection; The activation of glial cell and neuronic death have effect of equal importance; And exist between the two and necessarily get in touch (referring to Bales KR; Du Y, et al.The NF-kappaB/Rel family of proteins mediates Abeta-induced neurotoxicity and glial activation.Brain Res Mol Brain Res.1998; 57 (1): 63-72.).Behind inoculation removing amyloid-beta speckle, the nervous system lesion relevant with AD can be repaired.This fact provides strong evidence for the amyloid-beta theory.Other evidences of amyloid-beta theory comprise: trisomy 21 syndrome patient is because A β produces too much; AD appearance nervous system lesion can appear in early days; The only nervous system disease of its young patient becomes diffusivity A β deposition, explains that amyloid beta deposition possibly be neuro pathology's change the earliest; Behind some measures removing amyloids, nervous system lesion such as neuron shrinkage, microglial activation can reverse.The key link that these evidence explanation amyloid beta depositions are AD morbidities (referring to Zhou Huifang, Xue Bing, Wang Xiaomin; The sick progress of Alzheimer-amyloid-beta theory and main control strategy; The natural science progress, 2003,13 (2): 121-125.).

3, epidemiology situation

Along with developed country and the raising of developing country's average life span, the old number of suffering from dementia increases sharply.In all dementias, the ratio shared the highest (about 50% ~ 60%) with AD.

According to document announcement, with advancing age, under the influence of and gene aging in tissue, the incidence rate of dull-witted phenomenon constantly raises; Wherein, AD, rose one times from 60 years old with old rapid rising in per 4 years ~ 5 years; The sickness rate of AD accounts for 10% in the old people of one's mid-60s, in the old people more than 85 years old, account for 47%, has become No. four human killer.

The past academia thought once that China was the low dangerous country of AD, yet in second Chinese elderly dementia's disease of holding in the Tunxi, Anhui in the early time and depression scientific seminar and the international senile dementia scientific seminar, this viewpoint is challenged.Have 5 parts of research reports to show, China also is the high-risk country of AD.

Global at present patient's AD estimation is about 1,200 ten thousand, and the existing senile dementia patient of China is approximately 3,000,000～4,000,000, and is rapid ascendant trend, makes a definite diagnosis generally death in about 7 years ~ 9 years of back.Have data to show, China more than 60 years old population reached 1.2 hundred million as far back as 1998, and, be much higher than the rate of population increase with average annual 3.2% speed increase.Old man more than 75 years old has that the old man has 8,000,000 more than 2,000 ten thousand, 80 years old, and increases with average annual 5.4% speed, and middle period next century can reach about 400,000,000, will become the maximum country of the aged in the world.

Sino-U.S.'s Shanghai investigation in 1988 shows; China over-65s old people's dementia incidence rate is 4.6%; Wherein the proportion of AD is 63%, and MID is 28%, and the dementia that brain tissue impairment sequela, chemicals poisoning and nervous system disorder or malnutrition cause accounts for 10%.

The Epidemiological study in Shanghai shows that China 55 years old, the dull-witted prevalence of over-65s are respectively 2.57% and 4.6%.Senile disease medical research center, Beijing finds that to 2788 old man's more than 60 years old in Beijing area investigation have 208 old men to suffer from dementia, prevalence reaches 7.5%.Female patients 139 people wherein, prevalence reaches 9.7%, is significantly higher than male's 5.8%.Their research also shows, advanced age, low educational level, lives in the high risk factor that the rural area is a senile dementia.And PLA General Hospital shows 1728 retired veteran cadres' investigation, have in dull-witted family history, " Culture Revolution " be afflicted by persecution, electromagnetic field exposes, cerebral trauma also is the risk factor of senile dementia.In the investigation of Guangzhou City elderly dementia's prevalence more than 75 years old, the AD prevalence is 7.49%.

4, the medicine of senile dementia

AD is one of common complaint among the elderly, is a kind of gradual function of nervous system's degenerative deficiency disorder, and the cause of disease of primary disease is thrown a flood of light on as yet at present, is a thorny problem in the treatment always.

The research and development of antidementia agent has caused the great attention of countries in the world the world of medicine.In recent years, along with to the deepening continuously of aspect researchs such as old people's nervous physiology, biochemistry, pharmacology, cause the developmental research of related drugs constantly to make progress.Calendar year 2001 has 1270 multiple products to put on market, and 90% is the discovery eighties, the research and development success nineties in these products, and the new product of this type of drug development has outnumbered the number of any other treatment type drug development.

The medicine of treatment senile dementia mainly contains cholinesterase inhibitor at present, cerebral vasodilator, and calcium antagonists prevents A β deposition medicine, suppresses β, gamma secretase medicine and anti-inflammatory agent etc.Main application cholinergic drug or choline vinegar enzyme inhibitor are treated, and the medicine of drugs approved by FDA listing treatment AD is cholinesterase inhibitor.Be used for clinical cholinesterase inhibitor has tacrine, donepezil (donepezil) and huperzine first-class at present.This type of medicine is early stage and to improve cognitive dysfunction effective to dementia, but certain side effect is also arranged and act on shortcomings such as of short duration, and especially tacrine is serious to hepar damnification.Also often should require mental skill clinically and protect medicine such as calcium ion antagonist (nimodipine and the spirit of western pyrrole); Free radical scavenger (vitamin E, vitamin C and gingko leaf preparation) etc.But the curative effect at present clinical from said medicine is general, and major part all only is anti symptom treatment, can not improve mortality rate, and toxic and side effects is big, and effect is often not satisfactory.

Through nearly 10 years clinical showing; (be called for short: AChE) inhibitor has certain therapeutical effect to AD to Acetylcholinesterase; Its main medicine tacrine, sharp this bright, donepezil, galantamine have been the representative kind in this field, thereby have also promoted the development process of dementia treatment drug market.

5, the market development prospect of senile dementia

The world's seven big drug markets showed in 2000, and the AD drug market has increased nearly one times from more than 400,000,000 dollar of the mid-90, calendar year 2001 nearly 1,200,000,000 dollars of whole world AD market value; External analyst's prediction AD patient in following 10 years will be increased to about 20%, and the growth rate of AD drug market will have rising by a relatively large margin.

Because the patient is on the increase, the market sales revenue of AD medicine is also always in steady-state growth.The nineties, such medicine became the situation of selling well medicine, nineteen ninety-five world's sales volume reached 5,000,000,000 dollars.At the beginning of 21 century, the sales volume of such medicine has surpassed the share in the treatment cardiovascular disease treating medicine, treatment gastrointestinal disease medicine and the anti-infectives market that are arranged in front three, and its growth momentum is good.

At present; Such disease more and more receives the great attention of China national and society, and because such disease is similar to affluenza such as diabetes, equally need take medicine for a long time; Therefore its medication market enlarges along with aged tendency of population gradually, and market prospect is good.Based on this; For in time understanding the market situation of China treatment AD medicine; State food and drug administration south medication economics institute information centre chooses various schools of thinkers sampling hospital, 60 doctor experts and 120 consumers of China six main medication cities (Beijing, Shanghai, Guangzhou, Nanjing, Hangzhou, Chengdu), to China domestic senile dementia medication market (1999 and 2000) and mainly compete kind and carried out comprehensive market survey activity.

The treatment of AD accounts for the 7th of world's drug market, is worth 6,100,000,000 dollars to the end of the year 2005.And if present anti-AD drug main relief of symptoms is not contained the development of the state of an illness.Therefore; Working out a kind of efficacious therapy method, to stop potential pathogenic process be very necessary, thus it is considered herein that development is used to prevent, diagnose, detect, protect and the product of treating aspect such as AD particularly medicine can have remarkable social benefit and economic benefit.

Therefore; Traditional Chinese herbal medicine in conjunction with China; Therefrom excavate effectively, the medicine of low toxicity, inexpensive prevention, diagnosis, detection, protection, treatment and aspects such as research vascular dementia and directly related disease thereof, food, health product especially medicine and health product, have very important significance.

But, up to the present, do not find as yet the report of procyanidin B 2 phosphatide complexes as dementia product application facet through document retrieval etc.

Summary of the invention

The technical problem that will solve required for the present invention is to disclose a kind of procyanidin B relevant with effective ingredient in Chinese ₂Composition and method of making the same and purposes, to overcome the above-mentioned defective that prior art exists.

That is to say that one of the object of the invention is intended to clear and definite procyanidin B ₂Compositions, i.e. procyanidin B ₂Phosphatide complexes;

Two of purpose is clear and definite procyanidin B ₂The method for preparing of phosphatide complexes;

Three of purpose is clear and definite procyanidin B ₂The purposes of phosphatide complexes, i.e. the activity of dementia application facet, and then the procyanidin B 2 phosphatide complexes is used to prepare anti-angiogenic dementia product.

Described procyanidin B 2 is meant and comprises procyanidin monomers (comprising: catechin and epicatechin) through an active the strongest isomer in the dimer of polymerization formation that its structural formula is seen the Instructions Page 2 middle part.

Described dementia product is meant the product that is used to prevent, diagnose, detect, protect, treat and study dementia disease and directly related disease thereof; Be to comprise in medicine, reagent, food, the health product etc. one or more; Preferred agents and health product, further preferred health product.

Described dementia disease and directly related disease thereof are meant and comprise the vascular dementia disease; Alzheimer; Or in early stage, mid-term of the diseases such as mixing disease of vascular dementia disease and senile dementia and late period etc. one or more, the present invention has the early stage mild cognitive impairment of vascular dementia disease and Alzheimer, hypomnesis or symptom such as forgetful and prevents preferably and therapeutic effect.

The employed medicinal raw material of described procyanidin B 2 be meant procyanidin B 2 many natural plants of extensively existing; Like in Semen Vitis viniferae, Fructus Hippophae, black currant, cacao bean, Fructus Fragariae Ananssae, Semen Ginkgo, Semen arachidis hypogaeae or the Fructus Crataegi etc. one or more; Preferred Semen Vitis viniferae, Fructus Hippophae, Semen Ginkgo, Semen arachidis hypogaeae or Fructus Crataegi, further preferred Fructus Hippophae.

(1) technical conceive

The independent development original new drug is a present urgent task of China; Chinese medicine and pharmacy has a long history; Also accumulated rich experience with Chinese herbal medicine prevention and treatment disease aspect; Therefore directly from Chinese medicine the screen effective active component be a valid approach, also be the place of the advantage of Chinese original new drug development.

Procyanidin is the plant polyphenol compounds that is rich in the various plants, because its unique physiological hygiene function, and people's attention extremely in recent years.The procyanidin wide material sources extensively exist in many natural plants Semen Vitis viniferae, Fructus Hippophae, black currant etc.More abroad about STUDIES ON THE BIOACTIVITIES OF PROCYANIDINS; Because its extremely strong antioxidant activity; Procyanidin becomes the focus of research aspect anticancer, antiinflammatory, antioxidation, radioprotective, mutation, antiallergic, protection cardiovascular and anti-ageing the waiting for a long time of improving looks in recent years, and it is a kind of medicine that has potentiality for various diseases.Procyanidin is a kind ofly to gather the chemical compound that procyanidin (more than the pentamer) is formed by monomer (mainly being catechin, table catechu rope etc.), Oligomeric Proanthocyanidins (monomeric two, three, four polymeric acceptors) and height.Wherein dimer distributes the extensivelyst in all kinds of procyanidins, and antioxidant activity is the strongest, is most important one type.Dimer has multiple isomer because of 2 monomeric conformations or condensation key position difference, 8 kinds of isomers at present having identified, and procyanidin B 2 is a wherein active the strongest dimer in 8 kinds of isomers.

The inventor finds; The procyanidin B 2 phosphatide complexes can improve cerebral ischemia re-pouring mice and the permanent ligation rats with cerebral ischemia of bilateral common carotid arteries model learning memory ability; Thereby show that this compounds has the good curing effect to vascular dementia, dementia is showed mild cognitive impairment, hypomnesis or forgetful in early days also has therapeutical effect.

Dementia has become important death cause and the important social and the family burden of Chinese aging population; Development anti-dementia product; Especially prevent, diagnose, protect and treat the vascular dementia disease; Or the product of aspects such as early stage, mid-term of the diseases such as mixing disease of vascular dementia disease and senile dementia and late period particularly medicine have remarkable social benefit, economic benefit; The economic pressures and monitoring burden of family can be obviously alleviated, also patient's life quality can be obviously improved.

(2) procyanidin B 2 phosphatide complexes

One of the object of the invention provides a kind of phosphatide complexes of the procyanidin B 2 relevant with effective ingredient in Chinese.This procyanidin B 2 phosphatide complexes can improve the fat-soluble of procyanidin B 2 and transmembrane transport performance, increases its oral administration biaavailability, strengthens its pharmacologically active.

It is crude drug that the present invention adopts the water soluble ingredient procyanidin B 2, adopts suitable method for preparing to process the procyanidin B 2 phosphatide complexes with phospholipid; Or process peroral dosage forms such as tablet, capsule after further adding sorptive material etc.

Described procyanidin B 2 phosphatide complexes is made up of by certain mol proportion procyanidin B 2 and phospholipid, in suitable organic solvent, forms through reflux, washing drying;

Described certain mol proportion is l:10～15:l, and preferred proportion is l:5～5:l;

Described phospholipid is meant and comprises in natural phospholipid or the synthetic phospholipid etc. one or more, can be wherein a kind of independent use, also can be that two or more mixes and uses.

Described natural phospholipid is meant and comprises in soybean lecithin or the Ovum Gallus domesticus Flavus lecithin etc. one or more; Preferably soya lecithin;

Described synthetic phospholipid is meant and comprises in hydrogenated soy phosphatidyl choline, hydrogenated yolk lecithin, dipalmitoyl phosphatidyl choline, two palmityl PHOSPHATIDYL ETHANOLAMINEs, DSPC or the DSPE etc. one or more; In preferred hydrogenated soy phosphatidyl choline, hydrogenated yolk lecithin or the DSPE etc. one or more; Further preferred hydrogenated yolk lecithin;

The present invention has changed the physicochemical property of procyanidin B 2 through procyanidin B 2 and a certain amount of phospholipid are formed phosphatide complexes in suitable solvent, strengthens its fat-soluble and stability, improves its transmembrane transport performance, thereby improves its bioavailability.Preparation technology of the present invention is simple, favorable reproducibility, is fit to suitability for industrialized production.

(2) method for preparing of procyanidin B 2 phosphatide complexes

Two of the object of the invention provides a kind of method for preparing of phosphatide complexes of the procyanidin B 2 relevant with effective ingredient in Chinese.

The method for preparing of procyanidin B 2 phosphatide complexes of the present invention is: take by weighing an amount of procyanidin B 2 and phospholipid by certain mol proportion, join in the suitable organic solvent, under the certain reaction temperature conditions; After time, drying is removed organic solvent in certain reaction; Add atent solvent again and separate not compound free drug; Remove atent solvent, drying promptly gets the procyanidin B 2 phosphatide complexes;

Described organic solvent is to select for use to comprise in normal hexane, oxolane, methanol, ethanol, cyclohexane extraction, dioxy cyclohexane extraction, dichloromethane, ethyl acetate, chloroform, ether, acetone, n-butyl alcohol, dioxy six alkane, butanone or the isopropyl alcohol etc. one or more; That is: can be wherein a kind of independent use, also can be that two or more mixes and uses;

Preferably include in normal hexane, oxolane, ethanol, cyclohexane extraction, ethyl acetate, chloroform, ether, acetone, n-butyl alcohol or the isopropyl alcohol etc. one or more; Further preferred oxolane;

The consumption of described suitable organic solvent is counted drug level with procyanidin B 2 and is not less than lmL solvent/mg medicine, and the drug level of the best after the optimization is 5 ~ 40ml organic solvent/mg medicine.

Described reaction temperature is 10 ℃～90 ℃, preferred 60 ℃～90 ℃, and further preferred 78 ℃;

The described response time is 20min～8h, preferred 3h～8h; Further preferred 5.5h;

Described drying can adopt drying under reduced pressure or lyophilization, to remove organic solvent or atent solvent;

Described atent solvent can be selected for use and comprise in dichloromethane, chloroform, ethyl acetate or the oxolane etc. one or more, can be wherein a kind of independent use, also can be that two or more mixes and uses; Preferably include in chloroform or the oxolane etc. one or more, further preferred chloroform.

(4) activity of procyanidin B 2 phosphatide complexes and effect

The present invention has carried out many-sided test to the procyanidin B 2 phosphatide complexes in the activity of the aspects such as prevention, diagnosis, protection, treatment and research of vascular dementia disease or its directly related disease etc.

The test of procyanidin B 2 phosphatide complexes aspect prevention, diagnosis, protection and treatment vascular dementia disease

1, to the protective effect of vascular dementia due to the reperfusion injury of mice GBI

Group:

Blank group: give and the isopyknic normal saline of drug group (that is: positive controls, trial drug group).

Positive controls: give the medicine Folium Ginkgo.

The trial drug group: the procyanidin B 2 phosphatide complexes is low, and high 3 dose groups neutralize.

Method: with injected in mice anesthesia, the neck median incision separates the left and right side common carotid artery, the bulldog clamp blocking-up bilateral common carotid arteries blood flow some time.Removing bulldog clamp pours into again.Sham operated rats is not pressed from both sides bulldog clamp, and all the other steps are the same.Gastric infusion when mice is poured into again, sham operated rats and model group (that is: blank group) are irritated stomach and are given pure water.Mice is irritated stomach every day, carries out mice behind the continuous irrigation stomach 21d and keeps away dark test.Begin to carry out the mice water maze test when irritating stomach 23d.

The mice water maze test:

Form by the positive circular channels of a black lucite, have one in the tank, can supply mice to stand.Before the experiment, pour water in the labyrinth, the depth of water there was not platform, and water temperature is moderate.First day mice in training is placed in some seconds on the platform; Make its existence of understanding this safety zone, then with mice in order head put into water, timing from four quadrants successively towards tank; After mice was found platform and stops some seconds above that, timing stopped automatically.Removed platform on the 5th day, the swimming starting point of mice is two quadrantal points away from platform, and the platform number of times worn in the incubation period of mice platform and record in the record limiting time.

Mice is kept away dark laboratory method:

Experimental provision is a trained reflex case, is divided into light and shade two parts, and a small holes can have free passage mice in two Room, and the copper grid are arranged at the bottom of the case, and wherein the copper grid of bottom, darkroom pass to 36V voltage.During experiment mice is put into bright chamber and darkroom dorsad; Because of the mice dark and habit of getting into the cave in the family way,,, mice one will be shocked by electricity when entering into darkroom four-footed contact copper grid so can repeatedly pierce the darkroom at the experiment training period; This behavior is mistake, and its normal reaction manages to withdraw from the darkroom at once.

During first day adaptive training, the record mice gets into the number of times in darkroom and gets into the time of camera bellows for the first time in some minutes; Behind the adaptive training, open supply unit, mice is put into bright chamber and darkroom dorsad, after when mice one enters into darkroom four-footed contact copper grid, will being shocked by electricity, mice is taken out immediately.Do experiment after 24 hours again, the time that the interior mice of observation some minutes gets into camera bellows first is incubation period.

The result shows that the procyanidin B 2 phosphatide complexes has therapeutical effect to vascular dementia, improves its learning and memory function.

2, to the protective effect of the permanent ligation hemopoietic of bilateral common carotid arteries pipe dementia injury in rats

Blank group: give and the isopyknic normal saline of drug group.

Positive controls: give the medicine Folium Ginkgo.

The trial drug group: the procyanidin B 2 phosphatide complexes is low, and high 3 dose groups neutralize.

Animal: rat

Method: with rats by intraperitoneal injection anesthesia, the neck median incision separates bilateral common carotid arteries, pricks with toe-in, carries out gastric infusion then, and sham operated rats and model group are irritated stomach and given pure water.Rat is irritated stomach every day, and continuous irrigation stomach 21d carries out the rat water maze test.

The rat water maze test:

Form by the positive circular channels of a black lucite, have one in the tank, can supply rat to stand.Before the experiment, pour water in the labyrinth, the depth of water there was not platform, and water temperature is moderate.

First day rat in training is placed in some seconds on the platform; Make its existence of understanding this safety zone, then with rat in order head put into water from four quadrants successively towards tank, the timing some minutes; After rat was found platform and stops some seconds above that, timing stopped automatically.

Removed platform on the 5th day, the swimming starting point of rat is two quadrantal points away from platform, and the platform number of times worn in the incubation period of rat platform and record in the record certain hour.

The result shows that the procyanidin B 2 phosphatide complexes has therapeutical effect to vascular dementia, improves its learning and memory function.

Through experimentation, prove that all procyanidin B 2 phosphatide complexes of the present invention is a kind of medicine of safe, efficient, stable, inexpensive treatment of vascular dementia disease.

The procyanidin B 2 phosphatide complexes of addressing of the present invention can be with the administered of the compositions patient in the needs treatment, and said compositions comprises the procyanidin B 2 of treating effective dose and pharmaceutically acceptable carrier.

(5) purposes of procyanidin B 2 phosphatide complexes

1, general introduction

The purpose of this invention is to provide a kind of product that is used to prevent, diagnose, detect, protect, treat and study vascular dementia disease and directly related disease thereof; Comprise in medicine, reagent, the food etc. one or more; Preferred agents and health product, further preferred health product.

Completed acute toxicity testing proves, the mouse stomach administration surpasses 3.0g/kg to the maximum tolerated dose of this active site, shows that this single component is safe and reliable, solved that Chinese medicine ingredients is complicated, active constituent content is low and contained the problem of toxic component.Therefore, procyanidin B 2 phosphatide complexes and compositions thereof can be used for preparing anti-angiogenic dementia product, are medicine and the health product that feedstock production forms with procyanidin B 2 phosphatide complexes of the present invention preferably, further preferred health product.

2, the method for using of procyanidin B 2 phosphatide complexes and compositions thereof and requirement

Procyanidin B 2 phosphatide complexes of the present invention can be united use separately or with other active component; Comprise and be used to prepare the product that is used to prevent, diagnose, detect, protect, treat and study vascular dementia disease and directly related disease thereof; Comprise medicine, reagent, food etc., especially medicine.

Aspect concrete use, procyanidin B 2 phosphatide complexes of the present invention can use separately, can also use with other many chemical substances.These chemical substances biologically active or have the function of treatment disease whether no matter; Comprise miscellaneous function like collaborative amplification, antagonism or alleviate the side effect etc. of procyanidin B 2 phosphatide complexes, these chemical substances are to comprise in pharmaceutically acceptable carrier, food, natural product, chemical synthetic drug or the human medication etc. one or more; Preferably include in pharmaceutically acceptable carrier or the food etc. one or more; Further preferred pharmaceutically acceptable carrier.

" the pharmaceutically acceptable carrier " that this paper uses comprises one or more in any He all physiology suitable solvent, disperse medium, afterbirth, antibacterial and antifungal, isotonic agent or the absorption delay agent etc.The example of pharmaceutically acceptable carrier comprises one or more water, saline, PBS, glucose, glycerol or ethanol etc. and in the compositions one or more thereof.In many cases, in said composition, preferably include isotonic agent, for example, sugar, such as in the polyhydric alcohol of mannitol, sorbitol, sorbitol or the sodium chloride etc. one or more.Pharmaceutically acceptable carrier can also comprise a spot of auxiliary substance, one or more in wetting agent or emulsifying agent, antiseptic or the buffer etc. for example, and they have strengthened the effect duration or the effectiveness of this procyanidin B 2 phosphatide complexes.

See that from concrete classification said pharmaceutically acceptable carrier is meant the pharmaceutical carrier that the medicine and pharmacology field is conventional, comprises excipient, like in starch or the water etc. one or more; Lubricant is like in glycerol or the magnesium stearate etc. one or more; Disintegrating agent is like microcrystalline Cellulose etc.; Filler is like in starch or the lactose etc. one or more; Bonding agent is like in pregelatinized Starch, dextrin, cellulose derivative, alginate, gelatin or the polyvinylpyrrolidone etc. one or more; Osmotic pressure regulator is like in glucose, sucrose, sorbitol or the mannitol etc. one or more; Diluent is like water etc.; Disintegrating agent is like in agar, calcium carbonate or the sodium bicarbonate etc. one or more; Absorption enhancer is like quaternary ammonium compound etc.; Surfactant is like hexadecanol etc.; Absorption carrier is like in Kaolin or the soap clay etc. one or more; Lubricant is like in Pulvis Talci, calcium stearate, magnesium stearate or the Polyethylene Glycol etc. one or more; In addition, can also in compositions, add other adjuvant, like in flavouring agent or the sweeting agent etc. one or more.

For example; With the dissolving of active component procyanidin B 2 phosphatide complexes, suspendible or (for example be emulsifiable in the suitable aqueous solvent; In distilled water, normal saline or the Green's solution etc. one or more) or in the oil-based solvent (for example; Vegetable oil is one or more in olive oil, Oleum sesami, Oleum Gossypii semen, Semen Maydis oil or the propylene glycol etc. for example) in; Can make ejection preparation; Wherein can contain dispersant (for example, one or more in polyoxyethylene sorbitan monoleate, polyoxyethylene hardened castor oil 60, Polyethylene Glycol, benzyl alcohol, chlorobutanol or the phenol etc.), osmotic pressure regulator (for example, one or more in sodium chloride, glycerol, D9-mannose, D-sorbitol or the glucose etc.) in the solvent.In this case, if necessary, can add additive, for example solubilizing agent (for example, one or more in sodium salicylate or the sodium acetate etc.), stabilizing agent (for example, human serum albumin etc.), analgesic (for example, benzyl alcohol etc.) etc.

According to the invention and the procyanidin B 2 phosphatide complexes can also unite use with the form of compositions, particularly with other chemical substance such as medicine animal especially mammal is comprised that people or other animals treat compositions for use or similar compositions.Said mammal; Comprise in people, mice, rat, sheep, monkey, cattle, pig, horse, rabbit, dog, chimpanzee, baboon, Adeps seu carnis Rhiopithecus roxellanae, macaque or the Rhesus Macacus etc. one or more; In preferred people, mice, rat, monkey, pig, rabbit or the dog etc. one or more, one or more in further preferred people, rat or the monkey etc.For example, can with procyanidin B 2 phosphatide complexes of the present invention add be suitable for to curee's Pharmaceutical composition in.Usually, this Pharmaceutical composition comprises procyanidin B 2 phosphatide complexes of the present invention and pharmaceutically acceptable carrier.

The compositions of procyanidin B 2 phosphatide complexes particularly pharmaceutical composition can have various forms, comprises in the dosage forms such as liquid, semisolid and solid for example one or more; Wherein said pharmaceutical composition comprises that the procyanidin B 2 phosphatide complexes of treating effective dose is an active component, and one or more pharmaceutically acceptable carriers.

The pharmaceutical composition of procyanidin B 2 phosphatide complexes can adopt conventional production method well known in the art to process various dosage forms, and active component is mixed with one or more carriers, is made into required dosage form then.For example; Procyanidin B 2 phosphatide complexes of the present invention can further be processed dosage forms such as granule, capsule, tablet, pill through adding suitable adjuvant, and described adjuvant can be selected from one or more in starch, dextrin, lactose, silicon dioxide, calcium hydrogen phosphate, cyclodextrin, microcrystalline Cellulose, sodium carboxymethyl cellulose, magnesium stearate, the Pulvis Talci etc.

Described dosage form comprises one or more in tablet, capsule, granule, suspensoid, Emulsion, solution, syrup or the injection etc., takes one or more route of administration in oral or the injection (comprise in intravenous injection, intravenous drip, intramuscular injection or the subcutaneous injection etc. one or more) etc. to carry out prevention, diagnosis, detection, protection, treatment or the scientific research of vascular dementia disease and directly related disease thereof.

It is 0.5% ~ 99% active component procyanidin B 2 phosphatide complexes that pharmaceutical composition preferably contains weight ratio; Further preferably contain weight ratio and be 60% ~ 99% active component procyanidin B 2 phosphatide complexes, most preferably contain weight ratio and be 95% ~ 99% active component procyanidin B 2 phosphatide complexes.

The pharmaceutical composition of procyanidin B 2 phosphatide complexes generally must be aseptic and stable under the production condition of storage.Can said composition be mixed with solution, microemulsion, dispersion liquid, liposome or other is suitable for the ordered structure of high drug level.Through with a kind of of this procyanidin B 2 phosphatide complexes of aequum and required mentioned component or combine to add in the appropriate solvent and then carry out aseptic filtration and prepare aseptic parenteral solution.Generally speaking, prepare dispersion liquid through this procyanidin B 2 phosphatide complexes being added in the aseptic solvent that contains basic disperse medium and required above-mentioned other composition.Under the situation of the sterile powder that is used to prepare aseptic parenteral solution, the method for preparing of recommendation is vacuum drying and lyophilized preparation.For example, through passing through to keep required granular size such as the coating of lecithin, under the situation of dispersion liquid and, can keeping the adequate liquidity of solution through using surfactant.Through comprising that in said composition the medicament (for example Monostearate or gelatin) that postpones to absorb can reach the prolongation absorption of injectable composition.

When being used for the patient, procyanidin B 2 phosphatide complexes dosage of the present invention is 5 ~ 200mg/kgd, and this dosage or consumption decide according to the age of patient or user and the situation of body weight and health or patient's symptom usually.

Procyanidin B 2 phosphatide complexes of the present invention and Pharmaceutical composition thereof can comprise the procyanidin B 2 phosphatide complexes of the present invention of " treatment effective dose " or " prevention effective dose "." treatment effective dose " is meant at the dosage of necessity and effectively reaches the amount of required therapeutic effect under the time.The treatment effective dose of procyanidin B 2 phosphatide complexes can cause that at this individuality the factors such as ability of required reaction change according to the patient's condition, age, sex and body weight and this procyanidin B 2 phosphatide complexes such as individuality.The treatment effective dose also refers to that the useful therapeutic effect of this procyanidin B 2 phosphatide complexes surpasses the amount of its any toxicity or harmful effect." prevention effective dose " is meant the amount that under necessary dosage and time, effectively reaches required preventive effect.Because preventive dose is used for the ill preceding or early stage curee of disease, the prevention effective dose is usually less than the treatment effective dose.The typical non-limiting scope of the treatment of procyanidin B 2 phosphatide complexes of the present invention or prevention effective dose is 5 ~ 200mg/kg, more preferably 20 ~ 100mg/kg.Should note; Dose value will change according to disease type of desiring to alleviate and seriousness; That is to say when being used for the patient that procyanidin B 2 phosphatide complexes dosage of the present invention or consumption decide according to the age of patient or user and the situation of body weight and health or patient's symptom usually.In addition; Should understand; For any specific curee; Should along with the time according to individual need and give with or supervision give the professional judgement adjustment given dose system with the people of said compositions, and the dosage range that this paper sets is merely illustrative, the scope or the practice of compositions that can't the requirement for restriction protection.

That is to say, need be according to object, route of administration, institute's disease of treating and the situation etc. of treatment, change procyanidin B 2 phosphatide complexes of the present invention at every turn and/or dosage or the consumption of every day.For example, give mammal through vein, adult (like body weight 60kg) especially, the single dose of said procyanidin B 2 phosphatide complexes is about 50～200mg, preferably about 100mg, preferred administration every day 1～3 time.Can adjust dosage unit, so that best required reaction (for example, treatment or prevention are replied) to be provided.For example, can single heavy dose of administration can give several divided doses or reduce or increase dosage in proportion according to the urgency of treatment situation in a period of time.The non-intestinal compositions that preparation is easy to the unified dosage unit form of administration and dosage is especially favourable.The dosage unit form that this paper uses refers to be suitable for the physical separation unit of dosage unit of the mammalian subject of desire treatment; The calculating that each unit contains scheduled volume is used for together producing with required pharmaceutical carrier the active matter procyanidin B 2 phosphatide complexes of required therapeutic effect.The specification of dosage unit form of the present invention; Mixing the interior of this technology that is used for treating individual sensitivity procyanidin B 2 phosphatide complexes by the following specific characteristic of confirming and directly depending on following (a) this procyanidin B 2 phosphatide complexes and the particular treatment of desiring to reach or preventive effect with (b) in restriction.

3, the pharmaceutical dosage form of procyanidin B 2 phosphatide complexes and compositions thereof and route of administration

The product that is used to prevent, diagnose, detect, protect, treat and study vascular dementia disease and directly related disease thereof of procyanidin B 2 phosphatide complexes of the present invention and preparation of compositions thereof, wherein the product according to the requirement of beverage, food technology field preparation can be used in prevention, protection and treats vascular dementia disease and directly related disease thereof; Can be used in patient's treatment or health care according to the product of the requirement of medical technical field preparation; Can either directly be used to prepare the medicine of treatment or health care separately; Also can mix with many chemical substances or make up, directly or indirectly be used to prepare the medicine of treatment or health care.Chemical substance described here identical with described in this joint preceding text.

In the present invention, required material comprises raw material of the present invention, above-mentioned matching used chemical substance etc., all should adopt the material of food stage or pharmaceutical grade according to practical situation and needs.

Procyanidin B 2 phosphatide complexes of the present invention and compositions thereof can be used the whole bag of tricks administration known in the art, although route of administration/administering mode of in many therapeutic use, recommending is spray or oral administration.But the technical staff will appreciate that route of administration/administering mode changes with required result.In some practical implementation, this reactive compound can together prepare for example empty release formulation with the carrier of protecting this chemical compound to avoid rapid release, comprises that graft transmission system, transdermal paste one or more in transmission system or the microcapsule transmission system etc.In addition, can also use biodegradable, biocompatible polymer, for example one or more in ethylene-ethyl acetate, polyanhydride, polyglycolic acid, collagen protein, polyorthoesters or the polylactic acid etc.Prepare the equal patent applied for of many methods of this preparation or be generally those skilled in the art and know (referring to for example Sustained and Controlled Release Drug Delivery Systems; J.R.Robinson edits, Marcel Dekker, Inc.; New York, 1978).

Procyanidin B 2 phosphatide complexes of the present invention and compositions thereof, one or more modes in administered through oral or the parenteral etc. are applied to the patient who needs this treatment usually.

Be used for when oral, can be made into conventional solid preparation such as in tablet, powder, granule or the capsule etc. one or more.When implementing, procyanidin B 2 phosphatide complexes of the present invention can be together oral with for example inert diluent or assimilable edible carrier.This procyanidin B 2 phosphatide complexes (with its composition altogether, if desired) can also be wrapped in hard or soft shell gelatin capsules, is pressed into tablet or directly adds in curee's the meals.About oral therapeutic administration, can said procyanidin B 2 phosphatide complexes be added with excipient and uses with one or more forms in edible tablet, buccal tablet agent, lozenge, capsule, suspension, syrup or wafer or the like.

For to give procyanidin B 2 phosphatide complexes of the present invention outside the parenterai administration, possibly together give to this procyanidin B 2 phosphatide complexes coating or with this procyanidin B 2 phosphatide complexes with the material that prevents its inactivation.Can also the reactive compound that replenish be added in the said composition.Other medicine that in the specific implementation, procyanidin B 2 phosphatide complexes of the present invention and one or more can be used to treat disease is prepared altogether and/or is given altogether.Thisly unite use, can utilize this medicine that gives primely, therefore avoid possible toxicity or the complication relevant with various monotherapies than low dosage.

Process in liquid preparation such as water preparation, oil-suspending agent or other liquid preparation one or more, like in syrup, tincture or the elixir etc. one or more; When being used for parenteral, can be made in solution, water preparation or the oiliness suspending agent etc. of injection one or more.

In the above-described type of service, preferred form is one or more in tablet, coated tablet, capsule or the injection etc., further one or more in preferred tablet, capsule or the injection etc., preferred tablet especially.

In sum, procyanidin B 2 phosphatide complexes of the present invention and compositions thereof can be used for preventing, diagnose, detect, protect, treating and study the product of vascular dementia disease and directly related disease thereof, preferred agents and health product, further preferred health product.

(4) technological speciality

The present invention has expanded new medical usage to the procyanidin B 2 phosphatide complexes, also for prevention, diagnosis, detect, protection, treatment and research vascular dementia disease and directly related disease thereof provide a kind of new medicament sources.Procyanidin B 2 phosphatide complexes safety and low toxicity of the present invention; Pharmacological action is stronger; Its raw material sources are abundant, inexpensive, and preparation technology is simple, can be used for preparing the product of prevention, diagnosis, detection, protection, treatment and research vascular dementia disease and directly related disease thereof.

The prepared procyanidin B 2 phosphatide complexes of the present invention can significantly improve the fat-soluble of procyanidin B 2, changes its physicochemical property, strengthens it and penetrates biomembrane and get into sanguimotor ability, thereby significantly improve the procyanidin B 2 bioavailability.Preparation technology is simple simultaneously, is suitable for suitability for industrialized production.

The present invention studies the procyanidin B 2 phosphatide complexes targetedly, and the pharmacological action of procyanidin B 2 phosphatide complexes is stronger, and its raw material sources are abundant, and preparation technology is simple, and yield is high; And safe in utilization, particularly can one-object-many-purposes, brought into play effect to greatest extent, and the scope of application is wide especially, therefore apply easily, can have a tremendous social and economic benefits in the short period of time.

Procyanidin B 2 phosphatide complexes stable in properties; Use the quality of the pharmaceutical preparations of preparation stable; The effect of prevention, diagnosis, detection, protection, treatment and research vascular dementia disease and directly related disease thereof is obvious, so it is more suitable for preventing, diagnose, detect, protect, treating and study the suitability for industrialized production of vascular dementia disease and directly related disease product thereof.

In a word, active adaption of the present invention modern medical service and the job demand of scientific research field and the needs of human nature service, be the safe raw material that is used to prevent, diagnose, detect, protect, treat and study aspects such as vascular dementia disease and directly related disease thereof.

The specific embodiment

The present invention has studied the new pharmacological action of existing procyanidin B 2 phosphatide complexes and new purposes; A kind of raw material that can be used in preparation prevention, diagnosis, detection, protection, treatment and products such as research vascular dementia disease and directly related disease thereof is provided, has been convenient to the safe handling in medical industry and fields such as relevant industries such as food, beverage.

(1) test of procyanidin B 2 phosphatide complexes aspect prevention, diagnosis, protection and treatment vascular dementia disease

In order to set forth activity of the present invention and effect better, be described in detail through instance below, procyanidin B 2 sample wherein comes from Chinese market, and its specification is 95% for procyanidin B 2 content, and the content of procyanidin B 2 in phosphatide complexes is 25%.

1, to the protective effect of vascular dementia due to the reperfusion injury of mice GBI

Group:

Blank group: give and the isopyknic normal saline of drug group.

Positive controls: give the medicine Folium Ginkgo.

The trial drug group: the procyanidin B 2 phosphatide complexes is low, high 3 dose groups of neutralization.

Method: with the mouse peritoneal injecting anesthetic, the neck median incision separates the left and right side common carotid artery, bulldog clamp blocking-up bilateral common carotid arteries blood flow.Removing bulldog clamp pours into again.Sham operated rats is not pressed from both sides bulldog clamp, and all the other steps are the same.Gastric infusion when mice is poured into again, sham operated rats and model group are irritated stomach and are given pure water.After mice continuous irrigation stomach was some day, perfusion was carried out mice and is kept away dark test, mice water maze test again.

The mice water maze test:

Form by the positive circular channels of a black lucite, have one can supply mice to stand in the tank.Before the experiment, pour water in the labyrinth, the depth of water there was not platform.First day mice in training is placed on the platform, makes its existence of understanding this safety zone, then with mice in order head put into water from four quadrants successively towards tank, timing, after mice was found platform and stops above that, timing stopped automatically.Removed platform on the 5th day, the swimming starting point of mice is two quadrantal points away from platform, writes down the incubation period and the record of mice platform in 2 minutes and wears the platform number of times.

Mice is kept away dark laboratory method:

Experimental provision is a trained reflex case, is divided into light and shade two parts, and a small holes can have free passage mice in two Room, and the copper grid are arranged at the bottom of the case, and wherein the copper grid of bottom, darkroom pass to voltage.During experiment mice is put into bright chamber and darkroom dorsad; Because of the mice dark and habit of getting into the cave in the family way,,, mice one will be shocked by electricity when entering into darkroom four-footed contact copper grid so can repeatedly pierce the darkroom at the experiment training period; This behavior is mistake, and its normal reaction manages to withdraw from the darkroom at once.

During first day adaptive training, the record mice gets into the number of times in darkroom and gets into the time of camera bellows for the first time; Behind the adaptive training, open supply unit, mice is put into bright chamber and darkroom dorsad, after when mice one enters into darkroom four-footed contact copper grid, will being shocked by electricity, mice is taken out immediately.Do experiment again, the time that the observation mice gets into camera bellows first is incubation period, and the number of times that mice gets into camera bellows is an errors number.

The result shows that the procyanidin B 2 phosphatide complexes has therapeutical effect to vascular dementia, improves its learning and memory function.

2, to the protective effect of the permanent ligation hemopoietic of bilateral common carotid arteries pipe dementia injury in rats

Blank group: give and the isopyknic normal saline of drug group.

Positive controls: give the medicine Folium Ginkgo.

The trial drug group: the procyanidin B 2 phosphatide complexes is low, high 3 dose groups of neutralization.

Animal: rat

Method: with rats by intraperitoneal injection anesthesia, the neck median incision separates bilateral common carotid arteries, pricks with toe-in, and gastric infusion when rat is poured into again, sham operated rats and model group are irritated stomach and are given pure water.Behind some days of the rat continuous irrigation stomach, the rat water maze test is carried out in perfusion again.

The rat water maze test:

Form by the positive circular channels of a black lucite, have one can supply rat to stand in the tank.Before the experiment, pour water in the labyrinth, the depth of water there was not platform.

First day rat in training is placed on the platform, makes its existence of understanding this safety zone, then with rat in order head put into water from four quadrants successively towards tank, timing, after rat was found platform and stops above that, timing stopped automatically.

Removed platform on the 5th day, the swimming starting point of rat is two quadrantal points away from platform, and the platform number of times worn in the incubation period and the record of record rat platform.

More than the experiment and of the present invention other the experiment all the proof, procyanidin B 2 phosphatide complexes of the present invention with and pharmaceutical composition be a kind of safe, efficient, stable, the inexpensive treatment of vascular dementia disease and the medicine or the health product of directly related disease thereof.

The mensuration of saturated dissolubility, the mensuration of profit partition coefficient prove beneficial effect of the present invention and practicality in mensuration through saturated dissolubility in the water, the n-octyl alcohol below.

(2) mensuration of saturated dissolubility in the water of procyanidin B 2 and procyanidin B 2 phosphatide complexes

, add excessive procyanidin B 2 and procyanidin B 2 phosphatide complexes respectively and stir as solvent with water, centrifugal.Get the upper strata stillness of night and filter, get subsequent filtrate in volumetric flask, be diluted to standardize solution, survey ultraviolet absorptivity, triplicate then at the 265nm place.

Excessive procyanidin B 2 and phosphatide complexes thereof are dissolved in the water, reach dissolution equilibrium.Record other saturation solubility of branch.The procyanidin B 2 phosphatide complexes can significantly improve the water solublity of medicine, is original 2.6 times.

(3) mensuration of saturated dissolubility in the n-octyl alcohol of procyanidin B 2 and procyanidin B 2 phosphatide complexes

Respectively with water and n-octyl alcohol as solvent, add excessive procyanidin B 2 and stir, centrifugal.Get the upper strata stillness of night and filter, get subsequent filtrate in volumetric flask, be diluted to standardize solution, survey ultraviolet absorptivity, triplicate then at the 265nm place.

Excessive procyanidin B 2 and phosphatide complexes thereof are dissolved in the n-octyl alcohol, reach dissolution equilibrium behind the 3h.Record other saturation solubility of branch.The procyanidin B 2 phosphatide complexes can significantly improve fat-soluble 19 times more than of medicine.

(4) mensuration of the profit partition coefficient of procyanidin B 2 and procyanidin B 2 phosphatide complexes

The n-octyl alcohol solution of preparation procyanidin B 2 and procyanidin B 2 phosphatide complexes; Respectively get after part places distilled water to stir; Up and down two layers of solution separately, be diluted to suitable concentration at 165nm place the survey ultraviolet absorptivity, and according to equation 1. and standard curve 2. be converted into concentration.At last the n-octyl alcohol concentration at each pH value place is compared with concentration of aqueous solution, draw the profit partition coefficient.

The saturated solution of preparation procyanidin B 2; The n-octyl alcohol solution of preparation procyanidin B 2 phosphatide complexes.Precision pipettes the distilled water of n-octyl alcohol solution and different pH of each sample in round-bottomed flask, after jolting, measures as stated above.

Experiment shows that the water oil content distribution coefficient of procyanidin B 2 phosphatide complexes compares with procyanidin B 2, obviously increases, and indicates to be absorbed with in its body and possibly to improve.

(5) dosage form of procyanidin B 2 phosphatide complexes aspect the anti-angiogenic dementia product of preparation

The procyanidin B 2 phosphatide complexes can adopt method well known in the art to process various dosage forms; Like tablet, capsule, granule, suspensoid, Emulsion, solution, syrup, injection etc., take oral or route of administration such as injection (comprising intravenous injection, intravenous drip, intramuscular injection, subcutaneous injection), mucosa dialysis are carried out the control of vascular dementia disease.

When being used for the patient, dosage is according to the situation decision of patient's age, body weight, route of administration and symptom.

The present invention prepares injectable powder and generally adopts conventional freeze-drying, as solvent, the steps include: to get the procyanidin B 2 phosphatide complexes with water, adds excipient; Be dissolved in water, add active carbon, filtration sterilization, fill; Partly roll plug, lyophilization, tamponade are rolled lid and are got final product.Used excipient is selected from one or more in mannitol, gelatin hydrolysate, glucose, lactose, the dextran etc.Every bottle contains procyanidin B 2 phosphatide complexes 10 ~ 100mg.

The present invention prepares injectable powder also can adopt spray drying method, as solvent, the steps include: to get the procyanidin B 2 phosphatide complexes with water, adds or do not add excipient (excipient is the same); Be dissolved in water, add active carbon, filtration sterilization; Spray drying, aseptic subpackaged, tamponade is rolled lid and is got final product.Every bottle contains procyanidin B 2 phosphatide complexes 10 ~ 100mg.

When the present invention prepared small-volume injection, preparation got final product as solvent with water for injection, also can add appropriate amount of auxiliary materials, and adjuvant is selected from one or more in ethanol, propylene glycol, glycerol, Polyethylene Glycol, benzyl benzoate, the dimethyl acetylamide.Every contains procyanidin B 2 phosphatide complexes 10 ~ 100mg.

The present invention prepares glucose infusion liquid or sodium chloride transfusion; With water for injection as solvent; Add the preparation of an amount of glucose or sodium chloride and get final product, also can add appropriate amount of auxiliary materials, adjuvant is selected from one or more in ethanol, propylene glycol, glycerol, Polyethylene Glycol, benzyl benzoate, the dimethyl acetylamide.Every bottle contains procyanidin B 2 phosphatide complexes 10 ~ 100mg.

The present invention prepares oral formulations such as tablet, capsule, granule, oral liquid, and adjuvant can be lactose, starch, dextrin, stearate etc., presses the routine techniques preparation.

Method for preparing as for procyanidin B 2 is sophisticated method, delivers at public publications such as multiple periodicals and magazines and patent documentations, just repeats no more in the present invention.

In the present invention, the instance of the above-described specific embodiment and the following stated all is in order to set forth the present invention better, is not to be used for limiting scope of invention.

Further the present invention is described in detail through the mode of embodiment below.

One of method for preparing of embodiment 1, procyanidin B 2 phosphatide complexes

The procyanidin B 2 phosphatide complexes (medicine: fat=13:1)

Take by weighing procyanidin B 2 450mg and soybean phospholipid 50mg places conical flask, add 45ml.Oxolane in 40 ℃ of stirred in water bath 3h, forms transparent dark brown yellow solution, has the minute quantity medicine to separate out, and the decompressing and extracting organic solvent adds an amount of chloroform, and shake well removes by filter deposition, and filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

Two of the method for preparing of embodiment 2, procyanidin B 2 phosphatide complexes

The procyanidin B 2 phosphatide complexes (medicine: fat=6:1)

Take by weighing procyanidin B 2 800mg and soybean phospholipid 200mg places conical flask, add the 80ml. oxolane, in 40 ℃ of stirred in water bath 3h; Form transparent dark brown yellow solution, the decompressing and extracting organic solvent adds an amount of ethyl acetate; Shake well; Remove by filter deposition, filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

Three of the method for preparing of embodiment 3, procyanidin B 2 phosphatide complexes

The procyanidin B 2 phosphatide complexes (medicine: fat=2:1)

Take by weighing procyanidin B 2 600mg and soybean phospholipid 400mg places conical flask, add the 60ml. oxolane, in 40 ℃ of stirred in water bath 3h; Form transparent light brown yellow solution, the decompressing and extracting organic solvent adds an amount of dichloromethane; Shake well; Remove by filter deposition, filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

Four of the method for preparing of embodiment 4, procyanidin B 2 phosphatide complexes

The procyanidin B 2 phosphatide complexes (medicine: fat=l:1)

Take by weighing procyanidin B 2 400mg and soybean phospholipid 600mg places conical flask, add the 40ml. oxolane, in

40 ℃ of stirred in water bath 3h form transparent light brown yellow solution, and the decompressing and extracting organic solvent adds an amount of oxolane, and shake well removes by filter deposition, and filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

Five of the method for preparing of embodiment 5, procyanidin B 2 phosphatide complexes

The procyanidin B 2 phosphatide complexes (medicine: fat=l:2)

Take by weighing procyanidin B 2 245mg and soybean phospholipid 755mg places conical flask, add 24.5ml.Oxolane in 40 ℃ of stirred in water bath 3h, forms transparent light brown yellow solution, and the decompressing and extracting organic solvent adds an amount of chloroform, and shake well removes by filter deposition, and filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

Six of the method for preparing of embodiment 6, procyanidin B 2 phosphatide complexes

The procyanidin B 2 phosphatide complexes (medicine: fat=l:4)

Take by weighing procyanidin B 2 139mg and soybean phospholipid 86lmg places conical flask, add the 14mL oxolane, in 40 ℃ of stirred in water bath 3h; Form transparent light brown yellow solution, the decompressing and extracting organic solvent adds an amount of chloroform; Shake well; Remove by filter deposition, filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

Seven of the method for preparing of embodiment 7, procyanidin B 2 phosphatide complexes

The procyanidin B 2 phosphatide complexes (medicine: fat=l:10)

Take by weighing procyanidin B 2 60mg and soybean phospholipid 940mg places conical flask, add the 6mI. oxolane, in 40 ℃ of stirred in water bath 3h; Form transparent light brown yellow solution, the decompressing and extracting organic solvent adds an amount of chloroform; Shake well; Remove by filter deposition, filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

Eight of the method for preparing of embodiment 8, procyanidin B 2 phosphatide complexes

The procyanidin B 2 phosphatide complexes

Take by weighing procyanidin B 2 400mg and soybean phospholipid 600mg places conical flask, add the 40ml. oxolane, in 80 ℃ of stirred in water bath 3h; Form transparent light brown yellow solution, the decompressing and extracting organic solvent adds an amount of chloroform; Shake well; Remove by filter deposition, filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

Nine of the method for preparing of embodiment 9, procyanidin B 2 phosphatide complexes

The procyanidin B 2 phosphatide complexes

Take by weighing procyanidin B 2 400mg and soybean phospholipid 600mg places conical flask, add the 40ml. oxolane, in 40 ℃ of stirred in water bath 30min; Form transparent light brown yellow solution, the decompressing and extracting organic solvent adds an amount of chloroform; Shake well; Remove by filter deposition, filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

Ten of the method for preparing of embodiment 10, procyanidin B 2 phosphatide complexes

The procyanidin B 2 phosphatide complexes

Take by weighing procyanidin B 2 400mg and soybean phospholipid 600mg places conical flask, add the 40ml. oxolane, in 40 ℃ of stirred in water bath 12h; Form transparent light brown yellow solution, color wants dark, the decompressing and extracting organic solvent than mixing time is short; Add an amount of chloroform, shake well removes by filter deposition; Filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

11 of the method for preparing of embodiment 11, procyanidin B 2 phosphatide complexes

The procyanidin B 2 phosphatide complexes

Take by weighing procyanidin B 2 400mg and soybean phospholipid 600mg places conical flask, add 8m [.Oxolane in 40 ℃ of stirred in water bath 3h, forms transparent dark brown yellow solution, and the decompressing and extracting organic solvent adds an amount of chloroform, and shake well removes by filter deposition, and filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

12 of the method for preparing of embodiment 12, procyanidin B 2 phosphatide complexes

Procyanidin B 2 phosphatide complexes (egg yolk lecithin)

Take by weighing procyanidin B 2 400mg and egg yolk lecithin 600mg places conical flask, add the 40ml. oxolane, in 40 ℃ of stirred in water bath 3h; Form transparent light brown yellow solution, the decompressing and extracting organic solvent adds an amount of chloroform; Shake well; Remove by filter deposition, filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.13 of the method for preparing of embodiment 13, procyanidin B 2 phosphatide complexes

Procyanidin B 2 phosphatide complexes (hydrogenated phospholipid)

Take by weighing procyanidin B 2 400mg and hydrogenated phospholipid 600mg places conical flask, add the 40ml. oxolane, in 40 ℃ of stirred in water bath 3h; Form transparent light brown yellow solution, the decompressing and extracting organic solvent adds an amount of chloroform; Shake well; Remove by filter deposition, filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

14 of the method for preparing of embodiment 14, procyanidin B 2 phosphatide complexes

Procyanidin B 2 phosphatide complexes (ethyl acetate)

Take by weighing procyanidin B 2 400mg and soybean phospholipid 600mg places conical flask, add the 40ml. ethyl acetate, in 40 ℃ of stirred in water bath 3h; Form transparent light brown yellow solution, the decompressing and extracting organic solvent adds an amount of chloroform; Shake well; Remove by filter deposition, filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

15 of the method for preparing of embodiment 15, procyanidin B 2 phosphatide complexes

Procyanidin B 2 phosphatide complexes (chloroform)

Take by weighing procyanidin B 2 400mg and soybean phospholipid 600mg places conical flask, add the 40ml. chloroform, in 40 ℃ of stirred in water bath 3h; Form transparent light brown yellow solution, have small amount of drug to separate out, the decompressing and extracting organic solvent; Add an amount of chloroform, shake well removes by filter deposition; Filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.16 of the method for preparing of embodiment 16, procyanidin B 2 phosphatide complexes

Procyanidin B 2 phosphatide complexes (acetone)

Take by weighing procyanidin B 2 400mg and soybean phospholipid 600mg places conical flask, add 40mI. acetone, in 40 ℃ of stirred in water bath 3h; Form transparent light brown yellow solution, have medicine to separate out, the decompressing and extracting organic solvent; Add an amount of chloroform, shake well removes by filter deposition; Filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

17 of the method for preparing of embodiment 17, procyanidin B 2 phosphatide complexes

Procyanidin B 2 phosphatide complexes (dichloromethane)

Take by weighing procyanidin B 2 400mg and soybean phospholipid 600mg places conical flask, add the 40ml. dichloromethane, in 40 ℃ of stirred in water bath 3h; Because the solvent highly volatile forms transparent dark brown yellow solution, the decompressing and extracting organic solvent; Add an amount of chloroform, shake well removes by filter deposition; Filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.

18 of the method for preparing of embodiment 18, procyanidin B 2 phosphatide complexes

Procyanidin B 2 phosphatide complexes (methanol)

Take by weighing procyanidin B 2 400mg and soybean phospholipid 600mg places conical flask, add 40mI. methanol, in 40 ℃ of stirred in water bath 3h; Form transparent light brown yellow solution, have medicine to separate out, the decompressing and extracting organic solvent; Add an amount of chloroform, shake well removes by filter deposition; Filtrating is dry, promptly gets the procyanidin B 2 phosphatide complexes.Embodiment 19, procyanidin B 2 phosphatide complexes tablet

Prescription is formed: procyanidin B 2 phosphatide complexes: 0.5g (embodiment 5 methods make)

The skill process: the procyanidin B 2 phosphatide complexes is mixed a certain amount of micropowder silica gel, place mortar, porphyrize is crossed 80 eye mesh screens, and the powder of obtaining adds 0.5% magnesium stearate, and tabletting promptly gets.

Embodiment 19, procyanidin B 2 phosphatide complexes capsule

Prescription is formed: procyanidin B 2 phosphatide complexes: 0.5g (embodiment 5 methods make)

Technical process: the procyanidin B 2 phosphatide complexes is mixed a certain amount of starch place mortar, porphyrize is crossed 80 eye mesh screens, and the powder of obtaining is got the blank capsules shell, and is encapsulated, promptly gets.

Embodiment 20, procyanidin B 2 phosphatide complexes capsule

Prescription is formed: procyanidin B 2 phosphatide complexes: 0.5g (embodiment 5 methods make)

Technical process: the procyanidin B 2 phosphatide complexes is mixed a certain amount of dextrin place mortar, porphyrize is crossed 80 eye mesh screens, and the powder of obtaining is got the blank capsules shell, and is encapsulated, promptly gets.

Embodiment 21, procyanidin B 2 phosphatide complexes capsule

Prescription is formed: procyanidin B 2 phosphatide complexes: 0.5g (embodiment 5 methods make)

Technical process: the procyanidin B 2 phosphatide complexes is mixed a certain amount of sodium carboxymethyl cellulose place mortar, porphyrize is crossed 80 eye mesh screens, and the powder of obtaining is got the blank capsules shell, and is encapsulated, promptly gets.

The preparation of embodiment 22, procyanidin B 2 phosphatide complexes injection

Prescription:

Technology:

Get 20 ℃ of ethanol (95%) 2000g, add active carbon 5g and stir, through core group filtration under diminished pressure, subsequent use.Get recipe quantity procyanidin B 2 phosphatide complexes and add among the above-mentioned ethanol 2000g, be heated to about 40 ℃, fully stir; Make it to dissolve clear and bright, add the recipe quantity propylene glycol then, add fresh water for injection again and make full dose become 10000ml; Fully stir, mixing adds active carbon 3g; Clear and bright through core group filtration under diminished pressure, filtrating before and after merging is clear and bright with special-purpose core fine straining again.The qualified back embedding of inspection content and clarity, 100 ℃ of circulation steam sterilization 30min promptly get the injection that every 10ml contains procyanidin B 2 phosphatide complexes 30mg.

Embodiment 23, the test of procyanidin B 2 phosphatide complexes aspect prevention, diagnosis, protection and treatment vascular dementia disease

The purity of described procyanidin B 2 is 95%, and the content of procyanidin B 2 in phosphatide complexes is 25%.

1, to the protective effect of vascular dementia due to the reperfusion injury of mice GBI

Group:

Blank group: give and the isopyknic normal saline of drug group.

Positive controls: give medicine Folium Ginkgo (Dr Willmar Schwabe) 48mg/kg.

The trial drug group: the procyanidin B 2 phosphatide complexes is low, high 3 dose groups of neutralization are respectively 80mg/kg (containing procyanidin B 2 is 20mg/kg), 160mg/kg (containing procyanidin B 2 is 40mg/kg) and 320mg/kg (containing procyanidin B 2 is 80mg/kg).

Method: with kunming mice, 28～32g, with 3% chloral hydrate 10ml/kg intraperitoneal injection of anesthesia, the neck median incision separates the left and right side common carotid artery, bulldog clamp blocking-up bilateral common carotid arteries blood flow 30 minutes.Removing bulldog clamp pours into again.Sham operated rats is not pressed from both sides bulldog clamp, and all the other steps are the same.Gastric infusion when mice is poured into again, sham operated rats and model group are irritated stomach and are given pure water (0.1ml/10g).1 time/d of mouse stomach, continuous irrigation stomach 27d.Pouring into 21d again carries out mice and keeps away dark test.Pour into 23d again and begin to carry out the mice water maze test.

The mice water maze test:

Form by the positive circular channels of a black lucite (d=90cm), have one in the tank, can supply mice to stand.Before the experiment, pour water in the labyrinth, the depth of water there was not platform 2cm, 24 ± 1 ℃ of water temperatures.First day mice in training is placed in platform last 10 second; Make its existence of understanding this safety zone, then with mice in order head put into water from four quadrants successively towards tank, timing 2min; After mice was found platform and stops 10s above that, timing stopped automatically.Removed platform on the 5th day, the swimming starting point of mice is two quadrantal points away from platform, writes down the incubation period and the record of mice platform in 2 minutes and wears the platform number of times.

Mice is kept away dark laboratory method:

Experimental provision is a trained reflex case, is divided into light and shade two parts, and a small holes can have free passage mice in two Room, and the copper grid are arranged at the bottom of the case, and wherein the copper grid of bottom, darkroom pass to 36V voltage.During experiment mice is put into bright chamber and darkroom dorsad; Because of the mice dark and habit of getting into the cave in the family way,,, mice one will be shocked by electricity when entering into darkroom four-footed contact copper grid so can repeatedly pierce the darkroom at the experiment training period; This behavior is mistake, and its normal reaction manages to withdraw from the darkroom at once.

During first day adaptive training, the record mice got into the number of times in darkroom and gets into the time of camera bellows for the first time in 3 minutes; Behind the adaptive training, open supply unit, mice is put into bright chamber and darkroom dorsad, after when mice one enters into darkroom four-footed contact copper grid, will being shocked by electricity, mice is taken out immediately.Do experiment after 24 hours again, observing the time that mice in 3 minutes gets into camera bellows first is incubation period, and the number of times that mice gets into camera bellows is an errors number.

Keeping away dark result shows; Mice focal cerebral ischemia 2h causes the fourth ventricle in mice with vascular dementia model after pouring into 21d again; Model group learning and memory of little mouse function is impaired, and passive to keep away dark incubation period short and wear the case errors number and increase, and compares difference with sham operated rats and has remarkable meaning (P < 0.05); The middle and high dose groups of procyanidin B 2 phosphatide complexes can increase fourth ventricle in mice with vascular dementia and keep away dark incubation period; Basic, normal, high 3 dose groups all reduce wears the case number of times, compares with model group, and difference has significance meaning (P < 0.05).The result sees table 1.

Table 1, experimental fourth ventricle in mice with vascular dementia is kept away dark incubation period, worn influence (n=10, the x ± SD) of case number of times

Compare * P < 0.05 with model group

The water maze test result shows after mice focal cerebral ischemia 2h pours into 23d again and causes the fourth ventricle in mice with vascular dementia model; Model group learning and memory of little mouse function is impaired; The 5th day water maze time incubation period of undergoing training is compared remarkable increase with sham operated rats; Wear the platform number of times and significantly reduce (P < 0.05); The middle and high dose groups of procyanidin B 2 phosphatide complexes significantly reduces the time and the increase of fourth ventricle in mice with vascular dementia water maze escape latency and wears the platform number of times, compares with model group, and difference has significance meaning (P < 0.05).The result sees table 2.

Table 2, to experimental fourth ventricle in mice with vascular dementia water maze escape latency, the influence (n=10,

) of wearing the platform number of times

Compare * P < 0.05 with model group

The result shows that the procyanidin B 2 phosphatide complexes has therapeutical effect to vascular dementia, improves its learning and memory function.

2, to the protective effect of the permanent ligation hemopoietic of bilateral common carotid arteries pipe dementia injury in rats

Blank group: give and the isopyknic normal saline of drug group.

Positive controls: give medicine Folium Ginkgo (Dr Willmar Schwabe) 24mg/kg.

The trial drug group: the procyanidin B 2 phosphatide complexes is low, high 3 dose groups of neutralization are respectively 40mg/kg (containing procyanidin B 2 is 10mg/kg), 80mg/kg (containing procyanidin B 2 is 20mg/kg) and 160mg/kg (containing procyanidin B 2 is 40mg/kg).

Animal: SD rat

Method: with the SD rat, 220～250g is with 3% chloral hydrate 10ml/kg intraperitoneal injection of anesthesia; The neck median incision separates bilateral common carotid arteries, uses the Line 1 ligation; Gastric infusion when rat is poured into again (1ml/100g), sham operated rats and model group are irritated stomach and are given pure water.1 time/d of rat oral gavage, continuous irrigation stomach 30d.Pour into 21d again and carry out the rat water maze test.

The rat water maze test:

Form by the positive circular channels of a black lucite (d=150cm), have one in the tank, can supply rat to stand.Before the experiment, pour water in the labyrinth, the depth of water there was not platform 2cm, 24 ± 1 ℃ of water temperatures.

First day rat in training is placed in platform last 10 second; Make its existence of understanding this safety zone, then with rat in order head put into water from four quadrants successively towards tank, timing 1.5min; After rat was found platform and stops 20s above that, timing stopped automatically.

Removed platform on the 5th day, the swimming starting point of rat is two quadrantal points away from platform, writes down the incubation period and the record of rat platform in 1.5 minutes and wears the platform number of times.

The water maze test result causes the vascular dementia rats model after showing the permanent ligation 21d of rat bilateral common carotid arteries; The model group's learning and memory function is impaired; The 5th day water maze escape latency time of undergoing training is compared remarkable increase with sham operated rats, wear the platform number of times and significantly reduce (P < 0.05).The high, normal, basic dose groups water maze of procyanidin B 2 phosphatide complexes is hidden and is compared equal significance aspire to model group and shorten, and wears platform time number average more than model group, and high, normal, basic dose groups has been compared significant difference (P < 0.05) with model group.The result sees table 4.

Table 4, to experimental vascular dementia rat water maze escape latency, the influence (n=10,

) of wearing the platform number of times

Compare * P < 0.05 with model group

Above specifically experiment showed, procyanidin B 2 phosphatide complexes of the present invention and pharmaceutical composition are a kind of safe, efficient, stable, the inexpensive treatment of vascular dementia disease and the medicine or the health product of directly related disease thereof.

The mensuration of saturated dissolubility in the water of embodiment 24, procyanidin B 2 and procyanidin B 2 phosphatide complexes

The mensuration of saturated dissolubility, the mensuration of profit partition coefficient prove beneficial effect of the present invention and practicality in mensuration through saturated dissolubility in the water, the n-octyl alcohol below.

, add excessive procyanidin B 2 and procyanidin B 2 phosphatide complexes respectively and stir as solvent with 20ml water.In 1,2,2.5, respectively get 2ml centrifugal (3000r/min) 10min during 3h.Get the upper strata stillness of night and cross the 0.45um filter membrane, get subsequent filtrate 1ml, be diluted to standardize solution, survey ultraviolet absorptivity, triplicate then at the 265nm place in the 10ml volumetric flask.

Table 5, procyanidin B 2 and phosphatide complexes thereof the saturation solubility in water

Excessive procyanidin B 2 and phosphatide complexes thereof are dissolved in the water, reach dissolution equilibrium behind the 3h.Record other saturation solubility of branch, result's arrangement is (table 5) as above.The procyanidin B 2 phosphatide complexes can significantly improve the water solublity of medicine, is original 2.6 times.

The mensuration of saturated dissolubility in the n-octyl alcohol of embodiment 25, procyanidin B 2 and procyanidin B 2 phosphatide complexes

Respectively with the water of 20ml and n-octyl alcohol as solvent, add excessive procyanidin B 2 and stir.In 1,2,2.5, respectively get 2ml centrifugal (3000r/min) 10min during 3h.Get the upper strata stillness of night and cross the 0.45um filter membrane, get subsequent filtrate 1ml, be diluted to standardize solution, survey ultraviolet absorptivity, triplicate then at the 265nm place in the 10ml volumetric flask.

Table 6, procyanidin B 2 and phosphatide complexes thereof the saturation solubility in n-octyl alcohol

Excessive procyanidin B 2 and phosphatide complexes thereof are dissolved in the n-octyl alcohol, reach dissolution equilibrium behind the 3h.Record other saturation solubility of branch, result's arrangement is (table 6) as above.The procyanidin B 2 phosphatide complexes can significantly improve fat-soluble 19 times more than of medicine.

The mensuration of the profit partition coefficient of embodiment 26, procyanidin B 2 and procyanidin B 2 phosphatide complexes

The preparation content of dispersion is the procyanidin B 2 of 200 μ g/ml and the n-octyl alcohol solution of complex.Respectively getting 10ml, to place pH be 1.0,3.6,4.5,6.0, in 7.4 the 10ml distilled water, stir 3h under the room temperature after, up and down two layers of solution separately, be diluted to suitable concentration at 165nm place the survey ultraviolet absorptivity, and according to equation 1. and standard curve 2. be converted into concentration.At last the n-octyl alcohol concentration at each pH value place is compared with concentration of aqueous solution, draw the profit partition coefficient.

The saturated solution of preparation procyanidin B 2; Preparation is equivalent to the n-octyl alcohol solution of the phosphatide complexes of procyanidin B 2 200mg/ml concentration.Precision pipettes the distilled water of the different pH of n-octyl alcohol solution and 10ml of each sample of 10ml in the 50ml round-bottomed flask, jolt 3h after, measure as stated above.The result is (table 7 and table 8) as above.

The water oil content distribution coefficient of medicine is relevant with the distribution of medicine in biomembrane; The increase within the specific limits of water oil content distribution coefficient can make medicine get into biomembranous amount and increase, and helps the transmembrane transport of medicine; Get into intravital medication amount increase thereby make, bioavailability improves.Experiment shows that the water oil content distribution coefficient of procyanidin B 2 phosphatide complexes compares with procyanidin B 2, obviously increases, and indicates to be absorbed with in its body and possibly to improve.

The water oil content distribution coefficient of table 7, procyanidin B 2

The water oil content distribution coefficient of table 8, procyanidin B 2 phosphatide complexes

Claims (34)

1. a procyanidin B 2 phosphatide complexes is characterized in that, described procyanidin B 2 phosphatide complexes is made up of by certain mol proportion procyanidin B 2 and phospholipid, in suitable organic solvent, forms through reflux, washing drying;

Described certain mol proportion is l:10～15:l;

The structural formula of described procyanidin B 2 is following:

2. procyanidin B 2 phosphatide complexes according to claim 1 is characterized in that, described certain mol proportion is l:5～5:l.

3. procyanidin B 2 phosphatide complexes according to claim 1 is characterized in that, described phospholipid is meant and comprises in natural phospholipid or the synthetic phospholipid one or more.

4. procyanidin B 2 phosphatide complexes according to claim 3 is characterized in that, described natural phospholipid is meant and comprises in soybean lecithin or the Ovum Gallus domesticus Flavus lecithin one or more.

5. procyanidin B 2 phosphatide complexes according to claim 4 is characterized in that described natural phospholipid is a soybean lecithin.

6. procyanidin B 2 phosphatide complexes according to claim 3; It is characterized in that described synthetic phospholipid is meant and comprises in hydrogenated soy phosphatidyl choline, hydrogenated yolk lecithin, dipalmitoyl phosphatidyl choline, two palmityl PHOSPHATIDYL ETHANOLAMINEs, DSPC or the DSPE one or more.

7. procyanidin B 2 phosphatide complexes according to claim 6 is characterized in that, described synthetic phospholipid is meant and comprises in hydrogenated soy phosphatidyl choline, hydrogenated yolk lecithin or the DSPE one or more.

8. procyanidin B 2 phosphatide complexes according to claim 7 is characterized in that described synthetic phospholipid is a hydrogenated yolk lecithin.

9. according to each described procyanidin B 2 phosphatide complexes of claim 1 ~ 8, it is characterized in that described phospholipid can be wherein a kind of independent use, also can be that two or more mixes use.

10. according to the method for preparing of each described procyanidin B 2 phosphatide complexes of claim 1 ~ 8, it is characterized in that described this method for preparing is: take by weighing an amount of procyanidin B 2 and phospholipid by certain mol proportion; Join in the suitable organic solvent, under the certain reaction temperature conditions, in certain reaction after the time; Drying is removed organic solvent, adds atent solvent again and separates not compound free drug; Remove atent solvent, drying promptly gets the procyanidin B 2 phosphatide complexes.

11. the method for preparing of procyanidin B 2 phosphatide complexes according to claim 10; It is characterized in that described organic solvent is to select for use to comprise in normal hexane, oxolane, methanol, ethanol, cyclohexane extraction, dioxy cyclohexane extraction, dichloromethane, ethyl acetate, chloroform, ether, acetone, n-butyl alcohol, dioxy six alkane, butanone or the isopropyl alcohol one or more.

12. the method for preparing of procyanidin B 2 phosphatide complexes according to claim 11; It is characterized in that described organic solvent is to select for use to comprise in normal hexane, oxolane, ethanol, cyclohexane extraction, ethyl acetate, chloroform, ether, acetone, n-butyl alcohol or the isopropyl alcohol one or more.

13. the method for preparing of procyanidin B 2 phosphatide complexes according to claim 12 is characterized in that, described organic solvent is an oxolane.

14. the method for preparing of procyanidin B 2 phosphatide complexes according to claim 10 is characterized in that, the consumption of described suitable organic solvent is counted drug level with procyanidin B 2 and is not less than lmL solvent/mg medicine.

15. the method for preparing of procyanidin B 2 phosphatide complexes according to claim 14 is characterized in that, it is 5 ~ 40ml organic solvent/mg medicine that the consumption of described suitable organic solvent is counted drug level with procyanidin B 2.

16. the method for preparing of procyanidin B 2 phosphatide complexes according to claim 10 is characterized in that, described reaction temperature is 10 ℃～90 ℃.

17. the method for preparing of procyanidin B 2 phosphatide complexes according to claim 16 is characterized in that, described reaction temperature is 78 ℃.

18. the method for preparing of procyanidin B 2 phosphatide complexes according to claim 10 is characterized in that, the described response time is 20min～8h.

19. the method for preparing of procyanidin B 2 phosphatide complexes according to claim 18 is characterized in that, the described response time is 5.5h.

20. the method for preparing of procyanidin B 2 phosphatide complexes according to claim 10 is characterized in that, described drying can adopt drying under reduced pressure or lyophilization, to remove organic solvent or atent solvent.

21. the method for preparing of procyanidin B 2 phosphatide complexes according to claim 10 is characterized in that, described atent solvent can be selected for use and comprise in dichloromethane, chloroform, ethyl acetate or the oxolane one or more.

22. the method for preparing of procyanidin B 2 phosphatide complexes according to claim 21 is characterized in that, described atent solvent can be selected for use and comprise in chloroform or the oxolane one or more.

23. according to the application of each described procyanidin B 2 phosphatide complexes of claim 1 ~ 8 in preparation dementia product.

24. the application of procyanidin B 2 phosphatide complexes according to claim 23 is characterized in that, described dementia product is meant the product that is used to prevent, diagnose, detect, protect, treat and study dementia disease and directly related disease thereof.

25. the application of procyanidin B 2 phosphatide complexes according to claim 24 is characterized in that, described dementia product is to comprise in medicine, reagent, food or the health product one or more.

26. the application of procyanidin B 2 phosphatide complexes according to claim 25 is characterized in that, described dementia product is health product.

27. the application of procyanidin B 2 phosphatide complexes according to claim 24; It is characterized in that; Described dementia disease and directly related disease thereof are meant and comprise the vascular dementia disease; Alzheimer, or early stage, mid-term of the diseases such as mixing disease of vascular dementia disease and senile dementia or in late period one or more.

28. the application of procyanidin B 2 phosphatide complexes according to claim 27 is characterized in that, described dementia disease and directly related disease thereof are the early stage mild cognitive impairment of vascular dementia disease and Alzheimer, hypomnesis or forgetful.

29. the application of compositions in preparation dementia product according to each described procyanidin B 2 phosphatide complexes of claim 1 ~ 8.

30. the application of the compositions of procyanidin B 2 phosphatide complexes according to claim 29 is characterized in that, described dementia product is meant the product that is used to prevent, diagnose, detect, protect, treat and study dementia disease and directly related disease thereof.

31. the application of the compositions of procyanidin B 2 phosphatide complexes according to claim 29 is characterized in that, described dementia product is to comprise in medicine, reagent, food or the health product one or more.

32. the application of the compositions of procyanidin B 2 phosphatide complexes according to claim 31 is characterized in that, described dementia product is health product.

33. the application of the compositions of procyanidin B 2 phosphatide complexes according to claim 30; It is characterized in that; Described dementia disease and directly related disease thereof are meant and comprise the vascular dementia disease; Alzheimer, or early stage, mid-term of the diseases such as mixing disease of vascular dementia disease and senile dementia or in late period one or more.

34. the application of the compositions of procyanidin B 2 phosphatide complexes according to claim 33; It is characterized in that described dementia disease and directly related disease thereof are the early stage mild cognitive impairment of vascular dementia disease and Alzheimer, hypomnesis or forgetful.