CN104922181A - Preparation composition containing broccoli extract with effects of improving immunity and suppressing tumors and preparation method of preparation composition - Google Patents
Preparation composition containing broccoli extract with effects of improving immunity and suppressing tumors and preparation method of preparation composition Download PDFInfo
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- CN104922181A CN104922181A CN201410741566.XA CN201410741566A CN104922181A CN 104922181 A CN104922181 A CN 104922181A CN 201410741566 A CN201410741566 A CN 201410741566A CN 104922181 A CN104922181 A CN 104922181A
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Abstract
The invention provides an anti-tumor and immune-modulating composition, a preparation method and an application of the composition in preparing anti-tumor and immune-modulating medicines. The preparation composition is formed by mixing 1 to 20 parts of broccoli extracting solution, 0.5 to 1 part of chitosan oligosaccharide, 1 to 2 parts of ganoderan, 1 to 2 parts of cordyceps polysaccharide and 1 to 2 parts of dextrin in parts by weight; the preparation composition is characterized in that the content of sulforaphane is 2 to 15 mg/g and the content of polysaccharide is 200 to 400 mg/g in the preparation composition; the preparation method comprises the following steps: crushing, porphyrizing, filtering, hydrolyzing and macroporous resin adsorption separating broccoli seeds or corms to obtain a water solution containing sulforaphane, uniformly mixing the broccoli extracting solution, chitosan oligosaccharide, ganoderan, cordyceps polysaccharide and dextrin with water, spray drying into fine powder, preparing into granules, tablets, capsules and the like. Animals researches prove that the compound polysaccharide preparation has remarkable effects on improving immunity and suppressing tumors.
Description
Technical field
The invention belongs to functional food and field of medicaments, relate to and comprise Caulis et Folium Brassicae capitatae preparation and this extract and oligochitosan, ganoderan, the composite formula of Cordyceps polysaccharide and preparation method.
Background technology
In recent years, China's cancer morbidity is in rising trend, the research of various anti-cancer and cancer-preventing health food becomes current study hotspot, sulforaphen (Sulforaphane is rich in Caulis et Folium Brassicae capitatae, Sulforaphane), sulforaphen is that in all natural anti-cancer materials, effect is the strongest, the active component that effect is best, it has unique anticancer function, it can cause apoptosis and the cell block of cancerous cell, the II type detoxication enzyme in human body can be induced simultaneously, suppress the generation of I type enzyme simultaneously, the harmful components such as carcinogen and free radical are discharged eventually through multiple enzyme system, simultaneously, this composition can not remain in human body, body is had no side effect, it is a kind of novel anticancer component.
Oligochitosan (COS) be also Chitosan oligosaccharide, be a kind of water solublity that chitosan is obtained through method process such as enzyme hydrolysis or chemical method hydrolysis better, molecular weight is at the oligosaccharide of below 2KD.Oligochitosan overcomes the water-fast shortcoming of chitin, can be absorbed by the body through intestinal well.Large quantifier elimination proves, oligochitosan has good immunoloregulation function, and it can strengthen the phagocytic function, lymphopoiesis differentiation function etc. of mononuclear phagocyte NK cell and neutrophilic granulocyte; The Prostaglandin PGE2 that oligochitosan produces can strengthen chemotaxis (the 1.Suzuki K of neutrophilic granulocyte, Okawa Y, Hashimoto K, et al.Protecting effect of chitin and chitosan on experimentally inducedmurine candidiasis.Microbiol Immunol.1984; 28 (8): 903-912,2.Suzuki K, Tokoro A, Okawa Y et al.Enhancing effects ofN-acetyl-chitooligosaccarides on the active oxygen-generating andmicrobicidal activeties of peritoneal exuddate cells in mice.ChemPharm Bull (Tokyo) .1985:33 (2): 886-888).It also can Promote immunity cell cytokine production.Oligochitosan energy Promote immunity competent cell produces interleukin-1 and interferon.Oligochitosan also has obvious anti-tumor activity.Qinghua LIUs etc. find that oligochitosan can make S180 cell block in the G0/G1 phase, thus change the distribution (Qinghua LIU of S180 cell cycle; Hole celebrating Yan; Liu Hong; Oligochitosan is on the impact of S180 sarcoma cell apoptosis, cell cycle and apoptosis-related protein Bcl-2, Bax. Xuzhou Medical College's journal 2006,26 (4), 290 ~ 293.) apoptosis of oligochitosan also by regulating the expression of bcl-2 and Bax to induce LoVo and S180 cell.
Ganoderma is medicinal fungi, just thinks that it has effect of strengthening the body resistance, strengthening by means of tonics, extremely praises highly its property of medicine ancient times.Modern science detects and shows, Ganoderma is in immune adjustment, by strengthen host immune regulatory function reach antitumor action (Lin Zhibin. the progress [J] of Ganoderma anti-tumor activity and immunoregulation effect. Peking University's journal: medicine, 2002, 34 (5): 493-498.) ganoderan is main component in Ganoderma, be widely used in the health food improving immunity at present, large quantity research shows that ganoderan has anti-tumor activity simultaneously, may be (the Mao Jian etc. reached by motion and the adhesion properties of inhibition tumor cell, Food Science, 2010.V31.Nol, the progress of P295 ganoderan).
Cordyceps polysaccharide is the main effectiveness composition of Cordyceps, there is antitumor, immunomodulating, antioxidation, blood sugar lowering isoreactivity, because Cordyceps Resources scarcity price is expensive, major part healthy food material adopts Cordyceps fungus to cultivate through biofermentation, effective component extracting from mycelium, the Cordyceps mycelium polysaccharide comprising extraction has the effect identical with natural cs, and Cordyceps mycelium can resist or recover immunosuppressant that tumours of chemotherapeutic medicine causes or immunologic injury.Cordyceps mycelium physical ability promotes the biosynthesis of spleen DNA, increases the content of nucleic acid and protein, promotes the propagation of splenocyte; The increase of abdominal cavity adherent cell, Macrophage in Spleen, liver Kupffer Cell volume can be made, endochylema increases, caryoplasm loosens, intracellular acidic phosphatase activity strengthens and is state of activation, blood plasma carbon clearance rate regulating liver-QI, spleen phagocytic index can be significantly improved, have the dual regulation making immunologic function strengthen or weaken.After the research of Liu Minpei etc. shows that lotus euphorbia egg decoctum its mouse oral gives cultural mycelium continuously, spleen index is apparently higher than matched group, obviously can suppress the growth of mice with tumor in-vivo tumour, extend the life cycle of mice with tumor, main organs be showed no tissue pathologies change (Liu Minpei, Ma Shiying, Dai Bing. artificial cordyceps militaris fruiting body in tumor-bearing mice body tumor-inhibiting action research [J]. Shenyang army medicine, 1999,12 (2): 101-103.).
Our research shows that Caulis et Folium Brassicae capitatae containing sulforaphen and oligochitosan, ganoderan and Cordyceps polysaccharide have synergism, can improve the active anticancer of sulforaphen, and can play the effect of immunity moderation function.Not yet report that this several extract combination is improving the research report of immunity and anti-tumor aspect, also has no this kind of extract and prepares granule, tablet, the preparation technology of capsule and the report of Patents at present.
Summary of the invention
The object of the present invention is to provide a kind of raising immunity antineoplastic compositions, and preparation method, granule, tablet and capsule preparation method thereof.It is characterized in that, compositions is according to the ratio mixing composition of Caulis et Folium Brassicae capitatae extracting solution 1 ~ 5 part of weight, oligochitosan 0.5 ~ 1 part of weight, ganoderan 1 ~ 2 part of weight, Cordyceps polysaccharide 1 ~ 2 part of weight and dextrin 1 ~ 2 part of weight, it is characterized in that in said preparation compositions, sulforaphen content is 2 ~ 15mg/g, polyoses content is 200 ~ 400mg/g.
Above-mentioned composition preparation process: prepared by Caulis et Folium Brassicae capitatae, adopts broccoli seed, bud Seedling or bulb to be raw material.By raw material mechanical activation comminution, after defat with petroleum ether (petroleum ether: raw material=1: 1 weight ratio), add 3-5 times of water mixing, add proper amount of acetic acid, pH is adjusted to 5-6, add the compound enzyme (papain: cellulase=1: 1) to promote extracts active ingredients of amount of solid 1/100, control temperature 25-40 DEG C, hydrolysis 2-8 hour, glucosinolate in Caulis et Folium Brassicae capitatae is made to be converted into sulforaphen under myrosin effect, after reaction terminates, filters solid part, hydrolyzed solution is adsorbed by macroporous adsorptive resins, resin can select AB-8, D101, HP850, HP870 etc., after having adsorbed, the impurity adsorbed in resin is washed away respectively with three times of column volume deionized waters and 20% ethanol, with active component in 80% ethanol elution resin of three times of column volumes, eluent is through less than 20-40 DEG C vacuum concentration, obtain the solution of extract containing sulforaphen.
Above-mentioned Caulis et Folium Brassicae capitatae aqueous solution is according to the ratio of compositions noted earlier, be the paste liquid of 8-15% with oligochitosan, ganoderan, Cordyceps polysaccharide and the dextrin furnishing solid content that adds water, powder is made, inlet temperature 170-180 DEG C, leaving air temp 70-80 DEG C through spraying dry.Ganoderan is that Ganoderma mycelium extraction is obtained, and Cordyceps polysaccharide is also that Cordyceps mycelium extracts gained.
The preparation of composition granule: get above-mentioned Caulis et Folium Brassicae capitatae, oligochitosan, Cordyceps polysaccharide and ganoderan, adds starch or dextrin by the 20-40% of all extract gross masses, adopts bed process to make particulate matter, dry, obtains granule;
The preparation of composition capsule: get above-mentioned Caulis et Folium Brassicae capitatae, oligochitosan, Cordyceps polysaccharide and ganoderan, add starch or dextrin by the 20-40% of all extract gross masses, adopt bed process to make particulate matter, dry, load in capsulae vacuus, obtain capsule;
The preparation of composition tablet: get above-mentioned Caulis et Folium Brassicae capitatae, oligochitosan, Cordyceps polysaccharide and ganoderan, adds starch or dextrin by the 20-40% of all extract gross masses, makes particulate matter, and dry, tabletting, coating, obtains tablet.
Detailed description of the invention
Embodiment 1: the preparation of Caulis et Folium Brassicae capitatae
Get 3kg broccoli seed to pulverize through pulverizer, mixed defatting is carried out with 3kg petroleum ether, then petroleum ether is isolated, add 15kg water mix homogeneously, acetic acid is added in mixed solution, pH is transferred to 6, add 300 grams of mixed enzyme (papains: cellulase=1: 1), add deionized water 10kg, add appropriate vitamin C, keep pH to be 6, at room temperature (25 DEG C) stir the reaction that is hydrolyzed, the glucosinolate in broccoli seed is made to be converted into sulforaphen, hydrolysis 8h.Centrifugalize solid portion, hydrolyzed solution adds in vacuum concentration pot, is 0.06Mpa in vacuum, at 30 DEG C, extracting solution is concentrated into the half of original volume.Concentrated solution adds in the chromatographic column of the column volume 2L that Hp850 resin is housed, loading flow velocity 20ml/min, after resin absorption, washes away impurity with 6L deionized water, then washes the impurity adsorbed in resin with 6L20% ethanol.Finally use active component in 6L80% ethanol elution resin, vacuum concentration at 25 DEG C, reclaim ethanol, obtain Caulis et Folium Brassicae capitatae extracting solution, sulforaphen content 30g/L in extracting solution.
Embodiment 2: the preparation of composition grain
Caulis et Folium Brassicae capitatae extracting solution 1L in Example 1, mix with 0.5kg oligochitosan, 1kg ganoderan, 1kg Cordyceps polysaccharide and 1kg dextrin, add deionized water, be adjusted to the slurry that solid content is 10%, powder is made through spraying dry, inlet temperature 180 DEG C, leaving air temp 80 DEG C, obtains composition powder.Composition powder adds 20% starch, adopts fluidized bed granulation method to make particulate matter, dry, obtains granule.
Embodiment 3: the immunologic function test of compositions
Experimental technique: laboratory animal selects cleaning grade female mice (body weight 20g ± 2g), be divided into four groups, i.e. blank group, low dose group, middle dosage group and high dose group, the Caulis et Folium Brassicae capitatae compositions that mice is fed is the solution that composition grain prepared by embodiment 2 is dissolved in distilled water gained, basic, normal, high dosage group calculates by 5 times, 10 times, 30 times (people's recommended amounts is 2-4g/d) of people's recommended amounts, is respectively low dose group 300mg/kg, middle dosage group 600mg/kg, high dose group 1800mg/kg.Matched group gives equivalent distilled water.With reference to Ministry of Public Health " health food inspection and assessment technical specification " and document Xue Bin " immunotoxicology experimental technique " (Beijing Medical University, China Concord Medical Science University's combined publication, nineteen ninety-five publish) in method to mice carry out test on immune function carry out ConA inducing mouse Splenic vein hemodynamics test, measure mice serum hemolysin (Hemagglutination Method), antibody-producting cell detects, Turnover of Mouse Peritoneal Macrophages engulfs chicken red blood cell test (half intracorporal method), monocytes/macrophages detection of phagocytic function (mice carbonic clearance test method(s)), NK cytoactive detection (LDH algoscopy), its result is as follows:
Table 1 Caulis et Folium Brassicae capitatae compositions is on the impact (n=10, x ± s) of the mice spleen lymphocytes proliferation ability that ConA induces
| Group | Dosage (mg/kg) | Optical density difference |
| Blank group | 0 | 0.131±0.062 |
| Low dose group | 300 | 0.300±0.077 |
| Middle dosage group | 600 | 0.315±0.057 |
| High dose group | 1800 | 0.389±0.059 |
Comparing as shown in Table 1 by the post-stimulatory net added value of more each experimental group splenocyte ConA (representing with OD570 value) and negative control group, find that Caulis et Folium Brassicae capitatae compositions low dose group, middle dosage group, high dose group all can improve the multiplication capacity of the mouse spleen lymphocyte of ConA induction, difference all has significance (P < 0.05), and showing that Caulis et Folium Brassicae capitatae combination has stimulates splenocyte increment effect.
Table 2 Caulis et Folium Brassicae capitatae compositions is on the impact (n=10, x ± s) of mouse antibodies cellulation and serum hemolysin
| Group | Dosage (mg/kg) | Hemolysis plaque number/ | Anti-volumetric quantities |
| Blank group | 0 | 54±13 | 241±32 |
| Low dose group | 300 | 68±15 | 254±22 |
| Middle dosage group | 600 | 128±16 | 268±25 |
| High dose group | 1800 | 119±14 | 286±24 |
Note: hemolysis plaque number is the number of hemolysis plaque in 106 splenocytes.
The antibody-producting cell number (plaque number) of middle dosage group and high dose group is all higher than negative control group as shown in Table 2, and difference all has significance (P < 0.05).Show that Caulis et Folium Brassicae capitatae compositions has the value-added effect of enhancing antibody cellulation.Serum hemolysin measurement result shows: the Hemolysin product of middle dosage group and high dose group mice all has significance (P < 0.05) higher than negative control group difference.Show that Caulis et Folium Brassicae capitatae compositions has the effect of raising mice serum agglutinin level by promoting the generation of mice internal antibody thus improving humoral immune function.
Table 3. Caulis et Folium Brassicae capitatae compositions engulfs the impact (n=10, x ± s) of chicken red blood cell and carbonic clearance to Turnover of Mouse Peritoneal Macrophages
As can be seen from Table 3, the phagocytic rate of middle and high dosage group peritoneal macrophage all significantly raises compared with blank group with phagocytic index (P < 0.05).Show that Caulis et Folium Brassicae capitatae compositions improves the phagocytic activity of Turnover of Mouse Peritoneal Macrophages, carbon wheel is cleaned up phagocytic index each dosage group and is risen not fairly obvious.
Table 4. Caulis et Folium Brassicae capitatae compositions is to the activity influence (n=10, x ± s) of NK cells in mice
| Group | Dosage (mg/kg) | NK cytoactive % |
| Blank group | 0 | 25.5±10.55 |
| Low dose group | 300 | 32.4±12.15 |
| Middle dosage group | 600 | 35.6±11.32 |
| High dose group | 1800 | 38.8±10.33 |
As can be seen from Table 4 along with composition dosage increases, NK cytoactive increases, wherein middle and high dosage group NK cytoactive increases significantly (P < 0.05), illustrates that Caulis et Folium Brassicae capitatae compositions can improve the immunocompetence of mice.
Comprehensively analyzed by above-mentioned immunologic function experimental test result, the immunocompetence of middle and high dosage group mice significantly improves, and illustrates that Caulis et Folium Brassicae capitatae compositions has and improves immunization significantly.
Embodiment 4: Caulis et Folium Brassicae capitatae compositions is to Lewis lung cancer model mice tumor-inhibiting action
Test method: select SPF level mice 60, male, body weight 18-22g, tumor strain is the strain of Lewis lung cancer tumor, and Caulis et Folium Brassicae capitatae compositions is the granule of preparation in embodiment 1.
Modeling method: the Lewis lung cancer cell of trophophase of taking the logarithm, 200 mesh filter screens filter, and make single-cell suspension liquid, with normal saline, suspension are diluted to 6 × 10
6the cell suspending liquid of individual/ml, gets 0.1ml conventional method and is inoculated in mice right fore oxter, and after inoculation, mice is divided into lotus tumor model group, positive CTX matched group (CTX cyclophosphamide), low dose group, high dose group according to random digits table.CTX group is disposable celiac injection CTX solution 0.2ml (4mg/ml) after inoculating latter 72 hours, gavage 0.4ml every day (30mg/ml) after 72h after low dose group inoculation, gavage 0.4ml every day (90mg/ml) after 72h after high dose group inoculation, lotus tumor model group inoculation 24h after, every day gavage 0.4ml normal saline.Tumour inhibiting rate calculate according to document (Tian Fei etc., Fei Yi Wan to the expression of mice-transplanted tumor VEGF and Lung metastases correlation research [J] treatment and prevention of tumour magazine, 2004,11 (11): 1141-1143) calculate.
Experimental result is as follows:
Table 5. each group Lewis lung cancer mouse tumor weighs and tumour inhibiting rate compares (x ± s)
| Group | Mus number | Tumor heavy (g) | Tumour inhibiting rate (%) |
| Lotus tumor model group | 15 | 3.72±0.05 | --- |
| CTX matched group | 15 | 0.60±0.05 | 83.87 |
| Low dose group | 15 | 0.94±0.08 | 74.73 |
| High dose group | 15 | 0.62±0.05 | 83.33 |
As shown in table 5, high dose group and low dose group all have significant tumor-inhibiting action (P < 0.01) compared with lotus tumor model group, and wherein high dose group tumour inhibiting rate is more than 80%.Find out from the mice process of feeding, the survival condition of high dose group and low dose group mice and vigor, significantly better than CTX matched group, also illustrate that Caulis et Folium Brassicae capitatae compositions contributes to improving the immunity of mice.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the premise without departing from the principles of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (5)
1. one kind is improved immunity antineoplastic compositions, it is characterized in that, compositions is according to the ratio mixing composition of Caulis et Folium Brassicae capitatae extracting solution 1 ~ 5 part of weight, oligochitosan 0.5 ~ 1 part of weight, ganoderan 1 ~ 2 part of weight, Cordyceps polysaccharide 1 ~ 2 part of weight and dextrin 1 ~ 2 part of weight, feature is that in said preparation compositions, sulforaphen content is 2 ~ 15mg/g, and polyoses content is 200 ~ 400mg/g.
2. compositions preparation process: prepared by Caulis et Folium Brassicae capitatae, adopts broccoli seed, bud Seedling, bulb to be raw material.By raw material mechanical activation comminution, after defat with petroleum ether (petroleum ether: raw material=1: 1 weight ratio), add 3-5 times of water mixing, add proper amount of acetic acid, pH is adjusted to 5-6, add the compound enzyme (papain: cellulase=1: 1) to promote the extraction of effective ingredient of amount of solid 1/100, control temperature 25-40 DEG C, hydrolysis 2-8 hour, glucosinolate in Caulis et Folium Brassicae capitatae is made to be converted into sulforaphen under myrosin effect, after reaction terminates, filters solid part, hydrolyzed solution is adsorbed by macroporous adsorptive resins, resin can select AB-8, D101, HP850, HP870 etc., after having adsorbed, the impurity adsorbed in resin is washed away respectively with three times of column volume deionized waters and 20% ethanol, with active component in 80% ethanol elution resin of three times of column volumes, eluent is through less than 20-40 DEG C vacuum concentration, obtain the solution of extract containing sulforaphen.
Above-mentioned aqueous solution, according to the ratio of claim 1, is the paste liquid of 8-15% with oligochitosan, ganoderan, Cordyceps polysaccharide and the dextrin furnishing solid content that adds water, makes powder, inlet temperature 170-180 DEG C, leaving air temp 70-80 DEG C through spraying dry.
3. the preparation of granule: get above-mentioned Caulis et Folium Brassicae capitatae, oligochitosan, Cordyceps polysaccharide and ganoderan, adds starch or dextrin by the 20-40% of all extract gross masses, makes particulate matter, dry, obtains granule.
4. the preparation of capsule: get above-mentioned Caulis et Folium Brassicae capitatae, oligochitosan, Cordyceps polysaccharide and ganoderan, adds starch or dextrin by the 20-40% of all extract gross masses, makes particulate matter, dry, loads in capsulae vacuus, obtains capsule.
5. the preparation of tablet: get above-mentioned Caulis et Folium Brassicae capitatae, oligochitosan, Cordyceps polysaccharide and ganoderan, adds starch or dextrin by the 20-40% of all extract gross masses, makes particulate matter, and dry, tabletting, coating, obtains tablet.
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| CN111419857A (en) * | 2016-06-30 | 2020-07-17 | 深圳福山生物科技有限公司 | Glucoraphanin composition and application thereof |
| CN106963800A (en) * | 2017-04-12 | 2017-07-21 | 广州市天河华南发展有限公司 | A kind of Caulis et Folium Brassicae capitatae and its preparation method and application |
| CN106962918A (en) * | 2017-05-05 | 2017-07-21 | 甘肃寿鹿山药业有限公司 | A kind of Multifunctional protective health granules and preparation method thereof |
| CN108634319A (en) * | 2018-05-23 | 2018-10-12 | 宁波拜尔玛生物科技有限公司 | A kind of composition of adjunct antineoplastic and its application |
| CN108815463A (en) * | 2018-08-06 | 2018-11-16 | 李军 | Natural drug, preparation method and application with cancer prevention effect |
| CN109248171A (en) * | 2018-09-13 | 2019-01-22 | 黄泳华 | Composition containing Imatinib and plant polyose |
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Effective date of registration: 20161017 Address after: 100176 street Beijing economic and Technological Development Zone Branch No. 88 Yizhuang biomedical industry park E2 block 6 layer Applicant after: Beijing hi tech United Bioengineering Technology Research Institute Co Ltd Address before: 100102, room 206, block B, building 210, East Garden, Wangjing, Beijing, Chaoyang District Applicant before: Lin Qiang |
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