A kind of Grifola frondosa polysaccharide soft capsules and preparation method thereof
Technical field
The present invention relates to a kind of water-soluble active polysaccharide that utilizes Grifola frondosa fermentation mycelium or sporophore to extract---soft capsule that grifola frondosa beta-glucose is made and preparation method thereof.
Background technology
Grifolan is to be raw material with Grifola frondosa fermentation mycelium or sporophore, extracts through hot water extraction or cold alkali and thermokalite, and again through the ultrafiltration and concentration ethanol precipitation, the ion-exchange chromatography purification, lyophilization obtains, and also can adopt existing other technologies to make simultaneously.Grifolan contains albumen or does not contain albumen, and its monosaccharide consists of glucose or contains other monosaccharide.Wherein grifola frondosa beta-glucose is the ramose β of β-1-3-1-6-glucosan, glucose is formed by connecting by β-1-6 glycosidic bond and β-1-3 glycosidic bond in the molecule, and β-1-6 is connected to main chain, and β-1-3 is connected to side chain, contain albumen 1-2%, molecular weight 5-35 ten thousand dalton.Have good thermally-stabilisedly, its aqueous solution is in the time of temperature 20-125 ℃, and biological activity is not lost; When temperature is no more than 80 ℃, stable to alkali.
Above-mentioned grifola frondosa beta-glucose is a kind of biological response modifier, has the tumor of inhibition, immunomodulating, radioprotective and antiviral effect, can be used for the adjuvant drug of various tumors and immunodeficiency symptoms treatment.Such polysaccharide is a water-soluble substances, is insoluble to organic solvent and vegetable oil.At present, mostly grifolan is to be made into hard capsule or oral liquid, and the dosage form of making tablet is also arranged.
The content of polysaccharide is generally lower in the existing Grifola frondosa extract, about 15-20%, and wherein the content of beta glucan is lower, and dissolubility is about about 1% in the water, does not reach the basic content of biological activity performance.Based on above-mentioned reason, solid preparation or oral liquid all have the problem that effective component content is low, dose big, assimilation effect is poor, bioavailability is low.Therefore be badly in need of a kind of can reduce dose and be beneficial to swallow, can guarantee the grifolan preparation of polysaccharide intake again, remedy the deficiency of above-mentioned solid preparation or oral liquid.
Summary of the invention
At many weak points that prior art exists, the invention provides a kind of brand-new water-soluble active polysaccharide that utilizes Grifola frondosa fermentation mycelium or sporophore to extract---soft capsule that grifola frondosa beta-glucose is made and preparation method thereof.In this method, except that the polysaccharide that utilizes Grifola frondosa fermentation mycelium or sporophore itself to be contained as the active component, added high-load grifola frondosa beta-glucose again separately, improving the beta glucan content of whole preparation, and make soft capsule dosage form.Adopt the grifolan of this dosage form, its content height is easy to absorb, and bioavailability also is improved, and can be used for the auxiliary treatment of antitumor, radioprotective and other immunodeficiency symptoms, also can be used for the adjusting of body blood glucose, blood fat and blood pressure and the efficacy enhancing and toxicity reducing that chemicotherapy is treated.
Grifola frondosa polysaccharide soft capsules of the present invention, its proportioning is counted by weight: Grifola frondosa solid content 30-65 part, other additives 35-70 part.So promptly can guarantee the flowability of capsule 's content in capsule, can guarantee the content of effective ingredient Grifola frondosa solid content again, improve the drug effect of this soft capsule preparation.
No matter wherein, the Grifola frondosa solid content comprises grifolan and maitake mushroom mycelia or Grifola Frondosa sporophore, wherein when adopting the sort of hybrid mode, preferably keeps all containing grifolan, can guarantee that like this drug effect of medicine is in level preferably.In capsule 's content, the content of grifolan should be the 6%-20% of capsule 's content gross weight.
The Grifola frondosa polysaccharide soft capsules that the present invention relates to, wherein grifolan extracts the grifola frondosa beta-glucose of purification or the polysaccharide of α-configuration for process, and the main polysaccharide that adopts is a grifola frondosa beta-glucose.Also contain the part beta glucan in maitake mushroom mycelia or the sporophore.Contain Grifola frondosa glycopeptide and glycoprotein in addition, polypeptide wherein or albumen are to be connected with sugar chain with combined state, can make things convenient for the better absorption of human body.In the capsule 's content, when grifolan all adopts grifola frondosa beta-glucose, its content also should be the 6%-20% of capsule 's content gross weight, maitake mushroom mycelia is a fermentation mycelium, this mycelium and Grifola Frondosa sporophore are through micronizing, granularity gets final product greater than 100 orders, preferably adopts the 300-500 order, is beneficial to the abundant suspension of solid content in adsorbent.Above-mentioned grifolan can adopt existing method to make, and also can directly buy on market.
Other additive amount in the Grifola frondosa polysaccharide soft capsules are 35-70 part.This additive is made of adsorbent and additives; Wherein, what adsorbent generally adopted is edible vegetable oil, comprises soybean salad oil, edible peanut wet goods, also can be edible safflower seed oil, Testa oryzae oil or olive oil.It can be any one miscella of also forming more than two or three wherein.In order to reach best absorption dispersion effect, can be with soybean salad oil, edible peanut oil as a class adsorbent (I), edible safflower seed oil is as the independent adsorbent of a class (II), Testa oryzae oil and olive oil are as a class adsorbent (III), adopt the mixing of above-mentioned three class adsorbents to reach best, when adopting adsorbent (I) and adsorbent (II) to mix, its weight ratio is: 1-3:1 when mixing; When adopting the mixture of adsorbent (I), adsorbent (II) and adsorbent (III), its weight ratio is 4-7:2-4:1.
Additives are emulsifying agent or suspending agent or reinforcing agent or its mixture.
Emulsifying agent comprises a kind of in polyglycerin ester, sucrose ester, the unimolecule distillation glyceride or several, and its amount ranges is by weight the 0.5-5% into capsule 's content; Suspending agent can be made of Cera Flava, oil with hydrogenated soybean, Semen sojae atricolor oil emulsion and short chain linoleic acid, can also can be its whole mixture by any one or several compositions wherein, and its content range is by weight the 0.5-5% into capsule 's content.The adding of these auxiliary agents can make grifolan and other solid content better disperse in capsule, makes it obtain better utilization and improves drug effect, does not influence absorption of human body simultaneously, and human body is not had harm.
Reinforcing agent mainly has the material of potentiation to constitute by the activity to grifolan, as a kind of in Vc or Ve or the lecithin or several mixture, also can adopt the polysaccharide of other fungus polysaccharide or animal origin, as chitin, simultaneously also can be the vegetable oil in the adsorbent, as safflower oil.The content range of reinforcing agent is by weight the 0.5-5% into capsule 's content.After having adopted these reinforcing agents, the activity of grifolan is given full play to, and further be strengthened.The capsular drug effect of grifolan is greatly improved.
In above-mentioned adjuvant dicyandiamide solution, the part by weight of adsorbent and additives is 100:1-20.
When water soluble polysaccharide and other solid contents are made into enough little granule, can disperse uniformly and be suspended in the oil preparation system.Therefore as long as the content of polysaccharide is enough high in the extract, the soft capsule of making just contains the effective ingredient that is higher than other dosage forms.Liquid drug preparation, especially high concentration liquid preparation have more advantage than solid preparation.Because the good stability of grifolan itself, after having adopted above-mentioned dicyandiamide solution and proportioning, can be made into the soft capsule preparation that volume is little, content is high, not only can reduce dose and number of times, and be made into liquid condition and help absorbing, the characteristic of oil preparation more helps grifolan running and drug effect performance in vivo.Select for use suitable vegetable oil to make solvent, not only can improve the stability of system, also can further promote the action effect of grifola frondosa beta-glucose.In addition, good dicyandiamide solution can keep the high homogeneity of medicine, and soft capsule preparation also has canned accurate characteristics, can strict control dosage.
The method for preparing above-mentioned Grifola frondosa polysaccharide soft capsules comprises mixing of materials and capsule shaping, it is characterized in that: described mixing of materials may further comprise the steps:
(1) configuration of adsorbent: choose its mixed proportion of required edible vegetable oil by formula proportion and defer to above-mentioned principle, mix;
(2) layoutprocedure of dicyandiamide solution: adsorbent is heated to 60-90 ℃, adds emulsifying agent, suspending agent and reinforcing agent in turn, continue insulation according to proportioning;
(3) adding of solid content: under 60-90 ℃ of keeping warm mode, according to proportioning raw materials with Grifola frondosa solid content mix homogeneously, add in the above-mentioned adsorbent that contains emulsifying agent, suspending agent and reinforcing agent, the limit edged stirs, solid content is uniformly dispersed in liquid phase, after the adjuvant interpolation is finished, continue to stir the mixing that 30min finishes material.
When the reinforcing agent that adopts is solid, it can be added with the Grifola frondosa solid content, and not add in the adsorbent with emulsifying agent, suspending agent.
Capsule shaping can adopt existing forming technique, and concrete steps are as follows:
1, the material that mixes fully grinds by colloid mill under 60-90 ℃ keeping warm mode, and with 100 purpose screen filtrations, the feed liquid that grinds comes back in the colloid mill it, and circulation is carried out 2-3 time repeatedly, can be all by 100 purpose screen clothes up to material.
2, capsule pressing conditions, as the capsule press, 60-65 ℃ of colloid temperature, 45-55 ℃ of charging aperture temperature, mold temperature 40-45 ℃, rotating speed 5-10r/min, liquid paraffin is made lubricant, and air RH is 60%.
3, typing is cleaned and is dry, and the soft capsule that suppresses is finalized the design in rotating cage, and temperature 25-30 ℃, rotating cage rotating speed 5-10r/min, time 4h; 3-10min in the alcoholic solution of the capsule immersion 60-85% that typing is finished, drop removes ethanol after the gentle agitation, and this process repeats twice.Send into the hot blast chamber after the cleaning, 25-30 ℃ of temperature, rotating cage rotating speed 5-10r/min, air humidity 45%, continuous drying 18-24h.
After adopting this processing method, at first make solid content, especially the content of effective ingredient grifola frondosa beta-glucose improves greatly, has reduced dose when reaching effective dose.Solid content is carried out micronizing, and the use of emulsifying agent, suspending agent in addition makes its easier even suspension in the oiliness adsorbent, and loading amount is more accurate.In addition, the use of reinforcing agent has certain synergism to the activity performance of effective ingredient beta glucan.And as soft capsule dosage form, then have quantitatively accurately, be beneficial to and swallow, the short and bioavailability advantages of higher of disintegration time.
The Grifola frondosa polysaccharide soft capsules that adopts proportioning of the present invention and processing method to make, make by grifolan, grifola frondosa beta-glucose and maitake mushroom mycelia powder, its effective ingredient is greatly improved than existing grifolan preparation, it is beneficial to taking of patient and absorbs, the auxiliary treatment that can be used for antitumor, radioprotective and immunodeficiency symptoms also can be used for the adjusting of body blood glucose, blood fat and blood pressure and the efficacy enhancing and toxicity reducing that chemicotherapy is treated.
The specific embodiment
Embodiment 1
A kind of antitumor and immunoregulatory Grifola frondosa polysaccharide soft capsules, its used composition is by weight: grifolan and high-purity beta-Dextran 8, Grifola frondosa mycelia powder 32 (micronizing can be crossed 300 mesh sieves), safflower oil 38.5, olive oil 11, Testa oryzae oil 5.5, polyglycerin ester 1, Cera Flava 1.45, oil with hydrogenated soybean 2.05, vitamin C 0.5; Described grifolan is the crude polysaccharides that maitake mushroom mycelia or sporophore extract, and the weight ratio of itself and high-purity beta glucan is 1:9.
The preparation process of Grifola frondosa polysaccharide soft capsules:
1, measures safflower oil, olive oil and Testa oryzae oil in the prescription ratio, mix, as adsorbent;
2, adsorbent is heated to 60-90 ℃, adds polyglycerin ester, oil with hydrogenated soybean and Cera Flava successively, stir and make abundant thawing;
3, in the prescription ratio with solid adjuvant material mix homogeneously such as Grifola frondosa solid content and vitamin Cs, add in the adsorbent, the limit edged stirs, and makes to be uniformly dispersed.After all adjuvants interpolations are finished, continue to stir 30min;
4, the material that mixes fully grinds by colloid mill under 60-90 ℃ keeping warm mode, and with 100 purpose screen filtrations.The feed liquid that grinds comes back in the colloid mill it, and circulation is carried out 2-3 time repeatedly, can be all by 100 purpose screen clothes up to material;
5, clean and the drying process condition according to capsule compacting, typing, the soft capsule made according to the preparation method under the summary of the invention item, is progressively carried out subsequent treatment and makes capsule finished product.
Embodiment 2
A kind of antitumor and immunoregulatory Grifola frondosa polysaccharide soft capsules, its used composition is by weight: grifola frondosa beta-glucose 15, Grifola frondosa mycelia powder 25 (micronizing can be crossed 100 mesh sieves), safflower oil 38, soybean salad oil 12, Testa oryzae oil 5, polyglycerin ester 1, Cera Flava 1.45, oil with hydrogenated soybean 2.05, vitamin C 0.5.
The preparation process of Grifola frondosa polysaccharide soft capsules:
1, measures safflower oil, olive oil and Testa oryzae oil in the prescription ratio, mix, as adsorbent;
2, adsorbent is heated to 60-90 ℃, adds polyglycerin ester, oil with hydrogenated soybean and Cera Flava successively, stir and make abundant thawing;
3, in the prescription ratio with solid adjuvant material mix homogeneously such as Grifola frondosa solid content and vitamin Cs, add in the adsorbent, the limit edged stirs, and makes to be uniformly dispersed.After all adjuvants interpolations are finished, continue to stir 30min;
4, the material that mixes fully grinds by colloid mill under 60-90 ℃ keeping warm mode, and with 100 purpose screen filtrations.The feed liquid that grinds comes back in the colloid mill it, and circulation is carried out 2-3 time repeatedly, can be all by 100 purpose screen clothes up to material;
5, clean and the drying process condition according to capsule compacting, typing, the soft capsule made according to the preparation method under the summary of the invention item, is progressively carried out subsequent treatment and makes capsule finished product.
Embodiment 3
A kind of antitumor and immunoregulatory Grifola frondosa polysaccharide soft capsules, its used composition is by weight: grifola frondosa beta-glucose 10, Grifola Frondosa sporophore micropowder 25 (micronizing, can cross 500 mesh sieves), safflower oil 45, edible peanut oil 10, monoglyceride 1, Cera Flava 1.45, oil with hydrogenated soybean 2.05, vitamin C 0.5.
The preparation technology of present embodiment Grifola frondosa polysaccharide soft capsules adopts and embodiment 1 described identical preparation technology.
Experimental example:
Experimental example 1
The antitumor action of the Grifola frondosa polysaccharide soft capsules that employing embodiment 1 makes
Experimental animal is a white mice in the Kunming, and the experimental animal number average is 14 every group.Dosage is 60,120,240mg.kg
-1.d, continuous oral 5d, inoculation transplantation type tumor cell suspension is taken continuously after 15 days again and is measured, and the results are shown in Table 1.
The antitumor action of table 1 Grifola frondosa polysaccharide soft capsules
* this numerical value is increase in life span.By table 1 as can be known, Grifola frondosa polysaccharide soft capsules all has good inhibitory effect to murine sarcoma S180, hepatocarcinoma HepA solid type and Lewis lung cancer solid type transplantation tumor, and EAC ascitic type mice life is had to a certain degree prolongation effect.
Experimental example 2
The immunoregulation effect of the Grifola frondosa polysaccharide soft capsules that employing embodiment 1 makes
Experimental animal is a white mice in the Kunming, and the experimental animal number average is 14 every group.Dosage is 60,120,240mg.kg
-1.d, oral, measure after continuous 10 days, the results are shown in Table 2.
The immunoregulation effect of table 2 Grifola frondosa polysaccharide soft capsules
The result shows that the Grifola frondosa polysaccharide soft capsules per os gave mice 10 days, can improve toes swelling degree, antibody-producting cell number, carbon clearance phagocytic index and the monokaryon-macrophage phagocytic function of mice.Show that Grifola frondosa polysaccharide soft capsules has the effect that strengthens cellular immunization and humoral immune function.
Experimental example 3
The radiation resistance of the Grifola frondosa polysaccharide soft capsules that employing embodiment 1 makes
Experimental animal is a white mice in the Kunming, and the experimental animal number average is 34 every group.Grifola frondosa polysaccharide soft capsules dosage is 60,120,240mg.kg
-1.d, continuous oral 30d, each group is all with 8Gy
60The Co-radiation gamma once, irradiation back d2, d7 get half animal for every group and make numeration of leukocyte, second half observes mean survival time and 30d survival rate, the results are shown in Table 3, table 4.
Table 3 Grifola frondosa polysaccharide soft capsules is to the influence of murine interleukin sum
Table 4 Grifola frondosa polysaccharide soft capsules is to the influence of mice mean survival time and 30d survival rate
The result shows, behind the irradiation 2-7 days, the total number average of murine interleukin of taking Grifola frondosa polysaccharide soft capsules was higher than matched group; Take 240mg.kg
-1.d the mice of Grifola frondosa polysaccharide soft capsules, survival rate was significantly improved compared with the control in 30 days, and had the significance of difference.Above result shows, Grifola frondosa polysaccharide soft capsules can make that 30 days survival rates of mice obviously increase behind the irradiation, average survival rate prolongs, total white blood cells obviously increases.Illustrate that disposable irradiation has antagonism to Grifola frondosa polysaccharide soft capsules to higher dosage.