A kind of compositions and its application that contains sulfated fucan and propolis
Technical field
The invention discloses a kind of Pharmaceutical composition, and its application in adjusting immunity of human body.More particularly, the invention discloses a kind of sulfated fucan and propolis as the compositions of active component.
Background technology
Immune system is one of most important biosystem in human body, and its function has a significant impact human body.In case the function of immune system imbalance occurs, will cause human body that the disease resistivity is weakened.Immune disorder, immunologic hypofunction crowd ratio are increasing in recent years, will cause for a long time the disease of immune system patient sharply to increase in the past.Immune system is that human body is resisted the most important safeguard system of pathogen infringement, this system is by immune organ (bone marrow, thymus, spleen, lymph node, tonsil, little Peyer's patches, vermiform appendix etc.), immunocyte (lymphocyte, mononuclear phagocyte, neutrophilic granulocyte, basophil, eosinophilic granulocyte, mastocyte, platelet etc.), and immune molecule (complement, immunoglobulin, cytokine etc.) forms.It is the most effective weapon of body defence pathogen invasion, but the hyperfunction meeting of its function is to self organ or tissue's generation injury.In a lot of self-diseases, the not normal important pathogenesis often of function of immune system.Each component function of immune system be normally to keep the metastable assurance of body's immunity, the hyperfunction of the defective of any component or function all can be brought infringement to body.
Each component of immune system blazons whole body, and is intricate, and particularly immunocyte and immune molecule constantly produce in body, circulate and upgrade.Immune system has the discriminative power of height, can accurately identify own and non-own material, to keep the relative stability of body; Can also accept simultaneously, transmit, enlarge, store the information of immunity relevant to memory, reply and constantly adjust its responsiveness for immunologic information generation positive and negative, therefore the imbalance of function of immune system is totally unfavorable to human body: the identification ability of human body is extremely easy causes the generation of allergic phenomena (to use certain food, injectable drug anaphylaxis to occur, even cause shock), on the contrary can cause repeated infection; The Selfstabilizing ability of human body is abnormal, and immune system is reacted to the cell of self, causes autoimmune disease.This shows, the human immune system plays a part very important to the mankind's health, if its functional lability, human body probably can be by virus, these pathogen infringements of antibacterial.
Because immune system has so important effect, therefore seek a kind of nontoxic or low toxicity, effectively the medicine of adjusting immunity of human body becomes a research field that merits attention.
Sargassum is the important component part of marine living resources, and the Thallus Laminariae (Thallus Eckloniae) (Lam/nar/ajaponica aresch) of the brown sub-guiding principle Laminariales Laminariaceae Larminaria of Sargassum door is the highest Sargassum of output, and the Thallus Laminariae (Thallus Eckloniae) output of China occupies first place, the world.Thallus Laminariae (Thallus Eckloniae) claims again the ocean vegetable; contain abundant functional materials; trace element-selenium, cobalt, the chromium wanted as iodine, mannitol, vitamin (A, B, C, F), taurine and needed by human body; it is a kind of existing edibility; the macro product of medicinal, health care, industry, agronomical value is arranged again, all on the books in Compendium of Material Medica, " legendary god of farming book on Chinese herbal medicine ", " edible book on Chinese herbal medicine " and " Chinese medicinal herb compilation ".The polysaccharose substance that is rich in Thallus Laminariae (Thallus Eckloniae), its structure is totally different, different in kind, has complicated many-sided biological activity, has a extensive future in medicine, chemical industry, food service industry.
Sulfated fucan is a Sulfated polysaccharide of class, is present in Brown algae, is at first extracted from Thallus Laminariae (Thallus Eckloniae) with diluted acid in 1913 by Kylin.And be defined as the polysaccharide that contains a considerable amount of fucosees (fucose) and sulfate, or contain the polysaccharide of sulfo-α-L one fucose.Be called fucoidan (fucoidan or fucoidan, fucosan, sulfated fucan) according to international nomenclature principle unification now, also someone is called fucan and fucosan.Since extracting sulfated fucan first from Brown algae palmate Thallus Laminariae (Thallus Eckloniae), people have carried out a large amount of research to the sulfated fucan of separate sources.Now people have had comparatively clearly the composition of sulfated fucan and have understood, and it is a very complicated polysaccharide of class chemical composition and structure, take fucose and sulfate as main, along with the difference of algae also contains galactose, xylose, other compositions such as alduronic acid.Thallus Laminariae (Thallus Eckloniae) Fucoidan is by fucose.Galactose, xylose, alduronic acid, the monosaccharide such as arabinose form.Take fucose and galactose as main, both ratio is approximately 3:1.
Described sulfated fucan of the present invention can obtain in several ways, has had many pieces of documents and patent to relate to the preparation of sulfated fucan.Comprise that CN1129109A discloses by dried kelp soaking, for several times filtrations, secondary ethanol extraction, washing with alcohol, coordinated the alkali condensation method of adjusting PH scope etc.CN1344565A discloses another kind of preparation method, comprises the steps such as pretreatment of raw material, temperature control stirring lixiviate, centrifugal, concentrated, ethanol precipitation, dehydrated alcohol dehydration.CN1517356A is mixed with aqueous solution with fucoidan, adds hydrogen peroxide, hypochlorous acid or nitrous acid and salt thereof, and with the heating of gained mixed solution, the membrane ultrafiltration with molecular cut off 3000-5000 obtains fucoidan oligosaccharide.CN1670028A, CN1392160A, CN1197674A also disclose respectively methods such as adopting flocculation and have prepared Sargassum polysaccharides, and these preparation methoies are all the present invention's sulfated fucan acceptables used.
Impact (the Liaocheng University journal of kelp polysaccharide sulfate on Turnover of Mouse Peritoneal Macrophages activation and cytotoxicity, 2004.17 (2) 56-57) research thinks that kelp polysaccharide sulfate can activate Turnover of Mouse Peritoneal Macrophages, strengthen its immune function, opposing and kill tumor cell are had positive role.
It is that the laminarin of 15000D is to the cytophagous activation of mice that kelp polysaccharide sulfate has been studied a kind of mean molecule quantities such as L-fucose, D-galactose that comprise to the immunoregulation effect (Chinese immunology, 2000.2 (16)) of Turnover of Mouse Peritoneal Macrophages.Weigh with tumor control rate, inhibition tumor cell pathological changes effective percentage has reached 86.5%.Effectively activating macrophage of laminarin is described, secretory cell toxic effect molecule improves macrophage peroxidase activity and phagocytic function.
Two of flavone compound general references have a series of compound flavone compounds that the phenyl ring (A-and B-encircle) of phenolic hydroxyl group links mutually by central thricarbon atom.Often be connected with the functional groups such as phenolic hydroxyl group, methoxyl group, methyl, isopentene group in structure.In addition, it also normal and sugar be combined into glycosides.Most scientists think that the basic framework of flavone is produced by three malonyl coenzyme As and a cinnamyl coenzyme A biosynthesis.Experimental results show that through isotopic labeling the A ring comes from three malonyl coenzyme As, the B ring comes from the cinnamyl coenzyme A.
Flavone extensively exists in natural certain plants and berry, the nearly kind more than 4,000 of sum, and its molecular structure is not quite similar, and all belongs to flavone as rutin, hesperidin, quercimentin, green tea polyphenol, pattern glucosides, pattern thuja acid etc.The flavone of different molecular structures can act on the different organ of health, as Fructus Crataegi--and cardiovascular system, Lan Mei--eyes, tart fruit--urinary tract system, Fructus Vitis viniferae--lymph, liver, fruit of Ramulus Sambuci Williamsii--immune system.
Propolis (plopolis) is described as " purple gold ", is that Apis is used the resin that gathers from tender shoots and the trunk of plant and sneaks into self mandibular gland secretions and a kind of colloidal solid thing with aromatic odor that Cera Flava mixes.Propolis has stronger bacteriostasis, can treat various diseases, and has natural antioxygenic property and the immunologic function of enhancing body.The biology of propolis, pharmacology and medical functions are mainly relevant with contained flavone component in propolis.Propolis has the good reputation of " chromocor compound treasure-house ", contain more than 40 and plant chromocor compound, the flavone compound that now isolation identification has gone out from propolis has chrysin, apigenin, acacetin, Quercetin, galangin, rutin, kaempferide, rhamnetin, pinocembrin, 5,7 one dihydroxy one 3,4 one dimethyl flavone, 5 monohydroxies-4 and 7 one dimethoxy flavanones etc.But propolis source is different, and propolis flavone composition and content thereof are different, and flavone component is different from the place of production and collection season and notable difference arranged.The flavone that the present invention refers to is contained all flavone compounds in propolis.
The medical usage of flavone compound has been discussed in the research of total Flavones From Propolis and application (medicinal industry information, 2006.4.3 (11) 130-131), comprising the application of propolis flavone in improving body immunity.
Propolis flavone strengthens research (Yangzhou University's journal agricultural and life sciences version of immunization, 2006.3 (27) 21-23) PF (propolis flavone) can generate and peripheral lymphocyte proliferation by Promote immunity chickling antibody, promote the chicken spleen T lymphopoiesis of In vitro culture, and certain dose-effect and time-effect relationship are arranged.
Summary of the invention
The object of the present invention is to provide a kind of sulfated fucan compositions and in the application in adjusting immunity of human body field.The inventor studies and finds by with sulfated fucan, propolis or propolis or similar extract combination, adjusting immunity of human body to be had collaborative, outstanding effect, and its life-time service safety and effect all are better than existing product.
Creativeness of the present invention is to adopt sulfated fucan, as compositions, and uses it for adjusting immunity of human body in conjunction with propolis or its flavone extract.The present composition has not only kept the adjusting immunologic function of sulfated fucan itself, adds simultaneously propolis auxiliary adjustment effect.Consider that propolis also has the effect of antioxidation, removing free radical and slow down aging, the present composition has the suitability widely.When being applied to audient crowd, can be comprehensive improve to get their body constitution.Use the present invention and can regulate simultaneously humoral immunization, phagocyte, lymphsystem immunocompetence, compare with the medicine/health product of existing single adjusting humoral immunization on market, the effect of the present composition is omnibearing.Simple sulfated fucan medicine/health product and propolis focus mostly in the function of certain particular aspects of regulating health, causing on the one hand, user occurs puzzled when selecting product, the more important thing is that on the other hand medicine/the health product that derive from a plurality of different manufacturers can't obtain unified assurance at aspects such as safety, toxicity, consist of certain threat for the medication of consumer/edible safety.Evidence, compositions of the present invention can effectively improve body immunity and anti-fatigue ability, experimental results show that further the present invention is not only simultaneously safe and harmless, and its effect all has considerable raising with respect to independent sulfated fucan, propolis.
The present invention's propolis used can be not only the propolis of natural results, can be also it to be extracted the flavone extract of acquisition, openly analyze according to this area general knowledge and the health product composition that already gone on the market, those skilled in the art know that easily ginseng class plant extract, the Radix Astragali extracts such as Radix Ginseng, Radix Panacis Quinquefolii, Stichopus japonicus also can obtain the structure Flavonoid substances similar to propolis flavone to effect, so technical solution of the present invention has in fact also comprised the flavone component of these plants.Simultaneously, can directly extract flavone in propolis, also can take some simple extracting method to obtain propolis extract, make compositions such as wherein including Flavonoid substances, aromatic acid and ester, obviously, within this is encompassed in the scope of the invention.Technical scheme of the present invention has contained the compositions of sulfated fucan and propolis extract.
Technical solution of the present invention provides the compositions of a kind of sulfated fucan and propolis or its flavone extract, the propolis or its flavone extract that comprise the active component sulfated fucan and have obvious synergistic activity, sulfated fucan and propolis or propolis flavonoid ratio of weight and number can be 1:100-100:1, be preferably 1:30-30:1, most preferably be 1:10-10:1.
The inventor has studied simultaneously sulfated fucan, propolis or its flavone extract compositions has been added effect improved that vitamin C brings, applicant's experiment shows that vitamin C can further increase to effect of the present invention on the basis of the present invention's combination, therefore the present invention is also comprising such technical scheme: on the basis of the present composition, add appropriate vitamin C to improve implementation result of the present invention.
The present invention's sulfated fucan used seaweeds biology that has drawn from, comprise any plant that sulfated fucan can be provided, comprise Alga Sgrgassi Enerves, Thallus Laminariae, Sargassum fusiforme (Harv.) Setch, sargassum thunbergii, Thallus Sargassi Kjellmaniani, Thallus Laminariae (Thallus Eckloniae), yellow tang or Fucus Vesiculosus etc., preferred plant is Thallus Laminariae (Thallus Eckloniae), Alga Sgrgassi Enerves, be no matter artificial cultivation or spontaneous growth.The sulfated fucan method that obtains from these plants can be quoted the technical method described in background technology, and the molecular weight that obtains is on 100,000 order of magnitude levels.The sulfated fucan mean molecule quantity that obtains from Thallus Laminariae (Thallus Eckloniae), Alga Sgrgassi Enerves, Thallus Sargassi Kjellmaniani is respectively 350KD, 550KD, 980KD.When the present invention uses, can use the higher sulfated fucan of these molecular weight ratio, preferentially select the low-molecular-weight sulfated fucan of molecular weight between 5000D-200KD, low-molecular-weight algal polysaccharide sulfate used can extract from Sargassum also can be from the depolymerization of high molecular sulfated fucan.It is pointed out that the lasting progress along with technology, can drop in the scope of the invention for the sulfated fucan the change of molecular weight that the present invention uses.The purity of sulfated fucan used is not less than 50%, is preferably and is not less than 70%, more preferably is not less than 90%, and the best is to be not less than 95%.
Different according to algal kind used, there is certain difference in main group fucose, the sulfate content of the sulfated fucan that obtains, but this species diversity does not affect the present invention is applied to prevent and treat chemical liver injury, just reinforcement or the reduction of health-care effect that bring are within this belongs to the scope of the invention.The sulfated fucan of indication of the present invention contain chemically a class chemical substance of generally accepted sulfated fucan definition.
The present invention's flavone used can be that the flavone of purifying out in propolis can be also the preliminary extract of propolis that contains flavone component, can also make propolis medicinal material, propolis raw material etc.There has been kinds of schemes to disclose the method for extracting flavone.Chinese patent application CN2003101100850, CN031317529, CN031506488 etc. all disclose some methods of extracting the chromocor compound component from propolis.Propolis flavone Study on Extraction Method (Food Science, 2003.5 (24) 100-10) has been summarized the existing several technical matters that obtains flavone from propolis.
In the present invention, a preferred technique is that propolis raw material or propolis medicinal material are ground into coarse powder, carries out lixiviate according to the ethanol of the 50%-95% of the corresponding 5-10 times of quality of every g coarse powder, and solution temperature is controlled at 50-80 ℃, lixiviate 2-5 time, 1-3 hour at every turn.Merging filtrate, decompression and solvent recovery then, drying gets final product to get propolis flavone of the present invention.
The health product general reference that the present invention is directed to can improve and protect the product of health state, comprises F﹠B.The food of indication includes but not limited to Semen Tritici aestivi goods, starch-based product, meat product, plant extract, milk breast, health promoting candy etc.; The beverage of indication comprises that the drinkable flowing product of everyone body comprises common beverage and is used for improving the drinks of specific function, such as sports drink.
Compositions of the present invention can be combined with other nutrient substance or adjuvant.These nutrient substance comprise starch, dextrin, glucose, lactose, mannose, sucrose, microcrystalline Cellulose, syrup, several amino acids, calcium ferrum zinc etc.
Also can arrange in pairs or groups with adjuvant during present composition preparation, acceptable adjuvant comprises the guaranteed adjuvants of the upper safety of any pharmacy/threpsology such as excipient, antioxidant, PH regulator, antiseptic, isotonic agent.
The present composition can be prepared as polytype, comprises capsule, tablet, powder, solution, granule, sucks product etc., and the shape that changes health product belongs to coverage of the present invention.
The applicant analyzes present composition function, finds that regulating use amount of the present invention according to practical situation has positive role to adjusting immunity of human body.
The specific embodiment
Following embodiment is intended to illustrate specific embodiments of the present invention, does not limit protection domain of the present invention.In the following example, propolis flavone used is like this preparation: propolis medicinal material is ground into coarse powder, carries out lixiviate according to 80% ethanol of the corresponding 7 times of quality of every g coarse powder, solution temperature is controlled at 65 ℃, lixiviate 3 times, each 2 hours.Merging filtrate, decompression and solvent recovery then, drying gets final product to get propolis flavone of the present invention.
Preparation Example 1
Sulfated fucan 2mg
Propolis flavone 20mg
Starch 20mg
Mannitol 20mg
Glucose 15mg
Microcrystalline Cellulose 10mg
Silica gel 1mg
The mentioned component stirring and evenly mixing is become compositions, make capsule after gravimetry.
Preparation Example 2
Sulfated fucan 60mg
Propolis flavone 10mg
Starch 20mg
Glucose 15mg
Microcrystalline Cellulose 15mg
Silica gel 2mg
The mentioned component stirring and evenly mixing is become compositions, make buccal tablet.
Preparation Example 3
Sulfated fucan 25mg
Propolis flavone 2.5mg
Heme iron 7mg
Vitamin C 6ug
Folic acid 100ug
Mentioned component mixing grind into powder is dissolved in water to 500ml, seals, sterilization and packing, be prepared as the oral liquid that meets specific standard.
The present invention prepares oral liquid and can amplify on consumption in large-scale production, fully carry out after dissolving cannedly then sealing, sterilizing and be packed as oral liquid up to specification, and such as 500ml, 1L etc.
EXPERIMENTAL EXAMPLE 1
Experimental project: acute toxicity test
Experiment material: white mice is in a good state of health.The present composition, wherein sulfated fucan: propolis flavone mass ratio 3:1
Experimental technique:
Acute toxicity test: select 40 of white mice, each 20 of male and female, body weight is 18-22g.Each is divided into four dosage groups 16.00,7.27,3.31 at random according to the male and female mice, 1.50g/KG (bw), and Continuous Observation 15d after mouse stomach records toxicity performance and death condition.
Result of the test
Table 1 mice acute toxicity test result:
The health product of supposing production contrast with adult 60kg body weight daily intaking amount 0.05g/kg.bw the highest matched group that we adopt, 320 times of the intake that is equivalent to be grown up.The experimental observation fortnight has no poisoning symptom, and death toll is zero.Tested material is to the anxious toxicity LD of the mice of two kinds of sexes
50All greater than 15.00g/kg.bw. according to existing toxicity grading, belong to nontoxic rank.
EXPERIMENTAL EXAMPLE 2
Experiment material: select female mice, be divided at random four groups, body weight 18-22g.The present composition, wherein sulfated fucan: propolis flavone 3:1 is dissolved in deionized water and makes suspension.Cyclophosphamide for injection (cyclophospha-mide, Cy) is used for the hypoimmunity model construction.Sheep red blood cell (SRBC) (sheep e-rythrocyte, SRBC).
Experimental technique: according to human normal use amount adult 60kg body weight daily intaking amount 0.05g/kg.Bw, design is divided into high, normal, basic three dosage groups 0.25,0.10,0.05g/kg.Bw (dosage is equivalent to respectively 5,2,1 times of normal person's use amount), ten every group.Sulfated fucan matched group, propolis flavone matched group, model group, negative control group are set in addition, give respectively sulfated fucan, propolis flavone, distilled water, the distilled water of 0.10g/kg.Bw, be designated as respectively A matched group, B matched group, model group, matched group.Adopt gastric infusion, once a day.Measure every immune indexes after 20 days.Wherein in the time of the 15 day except negative control group the continuous 2d of every mouse peritoneal injection Cy50mg/kg (matched group waits the capacity normal saline), the animal model of preparation immunologic hypofunction. the every caudal vein of Ahau injects centrifugal rear 50% india ink (0.1ml/10g), get blood 401 from eyeball respectively after 2min and 10min, and it is added to 4ml0.1%NaHCO
3In solution, the 600nm wavelength is measured absorbance, presses literature method and measures phagocytic index.Record Mouse Weight, live in killing and get thymus and claim weight in wet base, with mouse thymus weight/body weight (mg/g) as thymus index.The assay method of half hemolysis value, lymhocyte transformation rate adopts conventional literature method.
The DTH method of testing is as follows: preparation DTH model, every Mus abdominal part was coated with 1%DNFB acetone Oleum Sesami solution 50ml sensitization in the 15 day, strengthen 1 time with dosage next day, prepare simultaneously immunodeficiency models except matched group, this model building method is with reference to above-mentioned model building method for test thymus index, phagocytic index.After sensitization three days, 1%DNFB solution evenly is applied in mouse right ear attacks.Attack 24h, crane one and put to death mice, cut the left and right auricular concha, lay round auricle with diameter 9mm card punch and weigh, the difference of left and right auricle is swelling.
The anti-fatigue ability test has nothing to do in immunity, therefore there is no model group, also need not to adopt the hypoimmunity model construction, only needs matched group to get final product.
The impact of 1 table 2 on mice delayed allergy DTH as a result
As a result 2: the impact of table 3 on normal mouse immune organ thymus index
As a result 3: the impact of table 4 on Function of Macrophages
As a result 4: the impact of table 5 on the mouse lymphocyte conversion ratio
As a result 5: the impact of table 6 on the mouse humoral immune ability
As a result 6: the impact of table 7 on the mouse anti-reflecting fatigue ability
Above-mentioned result of the test demonstration, the present composition has effectively improved the human body immunity.Contrast control experiment group data, the effect that the present invention improves immunity is better than using separately single component.Use the present composition and have the effect of regulating immunologic function, and this regulating action contained adjusting humoral system and phagocyte and lymphsystem, can omnibearing enhancing body immunity.Show: 1. omnibearing enhancing immunologic function under normal dose, comprise humoral immunization, phagocyte, lymphocytic immunity, and independent sulfated fucan or propolis flavone effect are so comprehensive.Such as propolis flavone one-component lymphocytic immunity effect is better, but humoral immunization, resisting fatigue poor effect.2. present composition unit's concentration is improved doses (such as 5 times of amounts), can suppress reinforced effects, perhaps reduce the human immunologic function.Illustrate that the present composition can the two-ways regulation immune function, to immunity is excited certain inhibitory action arranged under high dose.