CN104415132A - Compound formula for preparing soft capsule by pro-liposomes of ganoderma spore and freshwater mussel and preparation method thereof - Google Patents

Compound formula for preparing soft capsule by pro-liposomes of ganoderma spore and freshwater mussel and preparation method thereof Download PDF

Info

Publication number
CN104415132A
CN104415132A CN201310366989.3A CN201310366989A CN104415132A CN 104415132 A CN104415132 A CN 104415132A CN 201310366989 A CN201310366989 A CN 201310366989A CN 104415132 A CN104415132 A CN 104415132A
Authority
CN
China
Prior art keywords
extraction
pro
liposome
ganoderma spore
hbtdb
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
CN201310366989.3A
Other languages
Chinese (zh)
Inventor
宓鹤鸣
柳正良
胡水根
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
SHANGHAI SHENGCIKANG BIOMEDICAL TECHNOLOGY Co Ltd
Original Assignee
SHANGHAI SHENGCIKANG BIOMEDICAL TECHNOLOGY Co Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by SHANGHAI SHENGCIKANG BIOMEDICAL TECHNOLOGY Co Ltd filed Critical SHANGHAI SHENGCIKANG BIOMEDICAL TECHNOLOGY Co Ltd
Priority to CN201310366989.3A priority Critical patent/CN104415132A/en
Publication of CN104415132A publication Critical patent/CN104415132A/en
Pending legal-status Critical Current

Links

Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/06Fungi, e.g. yeasts
    • A61K36/07Basidiomycota, e.g. Cryptococcus
    • A61K36/074Ganoderma
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/56Materials from animals other than mammals
    • A61K35/618Molluscs, e.g. fresh-water molluscs, oysters, clams, squids, octopus, cuttlefish, snails or slugs
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/25Araliaceae (Ginseng family), e.g. ivy, aralia, schefflera or tetrapanax
    • A61K36/258Panax (ginseng)
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/48Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/48Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
    • A61K36/481Astragalus (milkvetch)
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/63Oleaceae (Olive family), e.g. jasmine, lilac or ash tree
    • A61K36/638Ligustrum, e.g. Chinese privet
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18Magnoliophyta (angiosperms)
    • A61K36/185Magnoliopsida (dicotyledons)
    • A61K36/75Rutaceae (Rue family)
    • A61K36/752Citrus, e.g. lime, orange or lemon
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/48Preparations in capsules, e.g. of gelatin, of chocolate
    • A61K9/4841Filling excipients; Inactive ingredients
    • A61K9/4858Organic compounds
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/33Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/37Extraction at elevated pressure or temperature, e.g. pressurized solvent extraction [PSE], supercritical carbon dioxide extraction or subcritical water extraction
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/30Extraction of the material
    • A61K2236/39Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
    • A61K2236/50Methods involving additional extraction steps
    • A61K2236/55Liquid-liquid separation; Phase separation

Abstract

The invention relates to a compound formula for preparing soft capsule by pro-liposomes of ganoderma spore and freshwater mussel and a preparation method thereof, which belong to the technical filed of traditional Chinese medicine, the formula is composed of ganoderma spore, freshwater mussel, gen-seng, astragalus membranaceus, ligustrum lucidum, pseudo-ginseng, fermented soybean and dried orange peel, has efficacy of supporting right and dispelling evil, increasing body immunity, inhibiting and killing cancer cell, preventing and treating cancer cell metastasis or recurrence, and can be used for various tumor patients, chemoradiotherapy patients, patients with immunocom promise, middle aged and old people, valetudinarianism and increased-immunity patients in office workers. According to the invention, effective parts in the formula medicinal material are separated and enriched to obtain lucid ganoderma total triterpenoid acid, mussels glycoprotein (HBTDB), total saponins and total flavonoids, content can reach more than 50%; and the invention also relates to a HBTDB liposome and pro-liposomes preparation and the method for assembling the pro-liposomes in the soft capsule. The preparation method employs the steps of supercritical CO2 fluid extraction, membrane separation, freeze drying and spray drying under vacuum, so that a novel dosage form of the soft capsule with the HBTDB pro-liposomes and other active site can be prepared.

Description

The formula of compound ganoderma spore mussel pro-liposome assembling soft capsule and method for making
Technical field
The present invention relates to technical field of Chinese medicines, be specifically related to a kind of prescription that can be used for treating effective ingredient in Chinese group compound preparation-" the compound ganoderma spore mussel pro-liposome assembling soft capsule " of carcinoma and control tumor chemoradiotherapy toxic and side effects, the preparation of the extraction and isolation of effective part group and enrichment, mussel pro-liposome and soft capsule preparation method.This product can be used in various tumour patient, immuuoeorapromised host, mid-aged population and working clan crowd weak and sickly, resistance descender.
Technical background
Cancer is the primary killers of harm humans life always, also wreaks havoc the whole world so far.The data issued the 21st the anticancer conference in the world according to International Union Against Cancer show, within 2008, there are 1,270 ten thousand people's cancer strickens in the whole world, and death toll is up to 7,600,000.The number of world wide endogenous cause of ill cancer mortality, adds up than acquired immune deficiency syndrome (AIDS), malaria and tuberculosis also many.If do not adopted an effective measure, expect the year two thousand thirty, every year by newly-increased for appearance 2,600 ten thousand cases of cancer, number of cancer deaths will reach 1,700 ten thousand, national " severely afflicated area " that will become cancer and wreak havoc of middle and low income.China national Minister of Public Health Chen Zhu points out in the 21st the anticancer conference opening ceremony speech in the world, and cancer has become the first cause of the death of Chinese city and urban residents.China's cancer incidence is in the quick rising stage, annual pathogenesis of cancer number increase about 2,600,000, dead 1,800,000 people.Current Chinese cancer spectrum has had the epidemic characteristic of developing country and developed country's cancer concurrently, and pulmonary carcinoma, hepatocarcinoma, gastric cancer, the esophageal carcinoma, colorectal cancer, leukemia, cerebroma, breast carcinoma, cancer of pancreas and osteocarcinoma are modal ten large cancer killers.
According to " 2012 Chinese tumors registration annual report " data that tumor Register national at the beginning of 2013 issues, the current whole nation just has 6 people to be diagnosed as cancer in every 1 minute, 8550 people are had every day to become cancer patient, annual new tumour patient of sending out reaches 3,500,000, and the sickness rate of China's tumor is 285.91/10 ten thousand.Cancer morbidity is not only increased sharply, and presents the trend of obvious rejuvenation, and 40 years old later pathogenesis of cancer presents one and significantly raise up.To this, national tumor Register deputy director Chen Wanqing represents, this is mainly due to the accumulation of nonage, and such as air pollution cannot embody in a short time for the harm of human body, but after 10 years even many decades, pathogenesis of cancer just can show.Wherein the cancer of most serious threat China residents ' health is exactly pulmonary carcinoma, dies from the patient of pulmonary carcinoma every year more than 600,000 people.This shows that China's prevention and treatment of malignant tumors situation is very severe, and malignant tumor has become serious social economical burden.Coming 10 years, cancer morbidity and the mortality rate of China will continue to rise.Associated specialist is analyzed, and aging is Fundamentals of formation of cancer.Along with viable organism is old and feeble gradually, cellular repair mechanisms falls into a decline, and particular cancers risk factor has also run up to higher degree, and therefore cancer morbidity also just significantly raises.Compare 2000, between 10 years, the tumor mortality rate of city, urban residents rises 14%, 41% respectively, reaches 1,68/,100,000,1,59/,100,000.
Since nearest 20 years, though China also achieves some significant progresses in treatment of cancer, but in treatment means except radiotherapy, chemotherapy, take except common drug analgesia, target treatment and excision, seem not have the Therapeutic Method of too positive effect.More worrying, these treatment meanss can make the misery of patient be growing on and on.In addition, some experts represent, cancer patient is very low to the ability of bearing complication, so the side effect for the treatment of is also very large problem, at this moment for patients, side effect is very painful.Cancer is difficult to cure, so that people talk " cancer " complexion changed, and the thick air of 21 century fear to cancer and hatred.In the face of the Emergency time that malignant disease is attacked, effective treatment of cancer becomes people's difficult problem the most urgently to be resolved hurrily.
Chinese traditional treatment tumor comes from the Zhou Dynasty the earliest, and just start there has been " cancer " word as far back as the Song dynasty, Chinese traditional treatment tumor has the history of very origin.Particularly nearly 30 years, Chinese traditional treatment tumor obtained the confirmation of a large amount of clinical and scientific researches, have also been obtained the generally accreditation of medical circle expert.Chinese medicine is anticancer to have one's own knack: Chinese medicine merges the toxic and side effects that Radiotherapy chemotherapy obviously can alleviate Radiotherapy chemotherapy, increase the curative effect of Radiotherapy chemotherapy, as general weakness, Nausea and vomiting, abdominal distention, appetite is bad and some hemogram decline, the toxicity of hepatic and renal function, and application Chinese medicine can improve; Postoperative patient applies Chinese medicine, can reduce postoperative relapse and metastasis.For some late tumors, due to physical weakness, be unsuitable for operation, also cannot carry out Radiotherapy chemotherapy, by application Chinese medicine, the development of tumor can be controlled, extend the life span of patient, improve the symptom of patient, improve the quality of life of patient.
Multiple anticancer active constituent is there is in Chinese medicine.The anticancer active constituent filtered out from higher plant in the world at present has tens thousand of kinds, also have in animal, fungus and marine organisms and there is a large amount of anticancer active constituents, so far belong to from more than 28 sections, in the Chinese herbal medicine of more than 3000 kinds, filter out the anticancer active constituent of more than 200 kinds.It is more much higher than screening hit rate in synthetic drug that basic research and clinical practice confirm from Chinese herbal medicine, carry out screening anticarcinogen.
TCM prevention and cure carcinoma has in mind from patient's animal economy, the class disease that cancer belongs to " vital QI being weakened and pathogen being violent, pathogen being excessive but vital QI weak ", so the basic principle for the treatment of is " strengthening vital QI to eliminate pathogenic factors, reinforcement and elimination in combination ".In conjunction with clinical datas such as medical history, the course of disease, comprehensive analysis of the data gained by the four methods of diagnosis and lab testings, comprehensively analyzing, determination for the treatment of based on pathogenesis obtained through differentiation of symptoms and signs, accomplishing " control real empty when turning round and look at, tonify deficiency does not forget reality ".The method of righting mainly according to the weakened body resistance difference stressed, and adopts QI invigorating respectively in conjunction with major lesions internal organs, enriches blood, nourishing YIN, YANG invigorating method for the treatment of; Eliminating evil employing mainly for pathological changes is regulated the flow of vital energy the methods such as circulation of qi promoting, dissipating phlegm and resolving masses, blood circulation promoting and blood stasis dispelling, heat-clearing and toxic substances removing.Meanwhile, patient had both noted after disease strengthening diet care, and harmonizing emotions, takes a good rest, and is conducive to the rehabilitation of cancer.
Chinese herb prevention disease, decoct from original medical material and be boiled into decoction to traditional cream, pellet, ball, loose, develop into the preparations such as solids numerous at present, liquid, injection, transdermal absorption system, targeting drug delivery system, all with better play drug effect, improve curative effect, reduce poisonous side effect of medicine directly related.Selection and the preparation of anti-cancer drug preparation dosage form then require higher, and wherein liposome (Liposomes) is different from an important feature of other ordinary preparation is that it has targeting.Liposome is as state-of-the-art at present, and the forth generation drug-supplying system being called " biological missile " becomes the novel form of targeting drug delivery system.Because liposome enters namely by the natural tendency that macrophage is engulfed as extraneous foreign body in body, therefore liposome is not only the ideal carrier of tumor chemotherapeutic drug, is also the ideal carrier of immunoactivator.
Inventor's previous research work basis is sturdy: Liu Zhengliang, the ring of tranquil crane, Hu Shuigen had once carried out applied basic research for many years to medicines such as key agents Ganoderma spore, mussel, Semen Sojae Preparatums in formula of the present invention respectively, form national inventing patent to have: " a kind of compound ganoderma lucidum spore soft capsule and preparation technology thereof " (numbering: ZL201110097796.3, Liu Zhengliang are inventor); " preparation method of clam lipoid and application thereof " (application number: 01126187.0, Authorization Notice No. CN1141083C, Hu Shuigen are the first inventor); Having studied Shanghai City biological medicine tackling of key scientific and technical problems problem 2, is the 03DZ that the ring of tranquil crane is presided over respectively,, the 03DZ19504 (research of Extract of Cyclina Sinensis HB-Z) that (research of Semen Sojae Preparatum) and Hu Shuigen preside over.Result of study shows: in Ganoderma spore triterpene acids and polysaccharide, mussel glycoprotein h B-Z and Semen Sojae Praeparatum extract, osajin all has the active anticancer suppressing kinds of tumor cells growth, the effect of body's immunity and nonspecific resistance and tool has a more substantial increase, clinical in needs chemicotherapy tumor patient, significantly can improve the toxic and side effects sent out chemotherapy and produce.
In view of above-mentioned technical background, we recognize that the outstanding theory of Chinese medicine and clinical practice experience have unique advantage on anti-curing cancers, develop cancer therapy drug and get twice the result with half the effort from natural Chinese medicine.The present invention is directed to the stem reality of the high incidence of current cancer, high mortality and high-band tumor existence colony clinical application outstanding problem, as narrow in the most antitumor spectra of existing listing antitumor drug, clinical selectivity is low, toxic and side effects is large, effective dose and toxic dose close, poor stability, the radiotherapy of tumor, chemotherapy are then simultaneously also kill normal cell in a large number pressing down tumor killing cell, thus cause Abwehrkraft des Koepers extremely to decline, cannot completed treatment process.Therefore, treatment of cancer is in the birth of calling low-poison efficient antineoplastic medicine, particularly from Chinese Traditional Medicine treasure-house, excavate anticancer complex prescription with Chinese characteristics, in conjunction with new and high technology, embody the modernization of Chinese medicine, the prescription playing Chinese medicinal formulae is rigorous, prevention of diseae, treating both the principal and secondary aspects of a disease, multicomponent is collaborative, efficacy enhancing and toxicity reducing, multiaction target spot, multichannel approach etc. advantage.Adopt liposome and pro-liposome novel form simultaneously, strengthen the targeting to treatment of cancer.The present invention has developed a kind of modern novel formulation of Chinese medicine of resisting carcinoma---" compound ganoderma spore mussel pro-liposome assembling soft capsule ".
Summary of the invention
This problem with Ganoderma spore and mussel for monarch drug, new and high technology such as application supercritical fluid extraction etc. is extracted and triterpenes anti-cancer effective component (A) and the ganoderan active site (B) such as Ganodenic acid in enrichment Ganoderma spore, apply freezing homogenate, water extract-alcohol precipitation, saltout and the anticancer effective site of glycopeptide class (C) in the technological means separation and concentration mussel such as lyophilization, this glycoprotein position reverse phase evaporation is prepared into liposome, add suitable protective agent, adopt freeze-drying to obtain this lipid freeze-dry powder (D); According to Chinese medicine traditional outstanding, theoretical and experience is for instructing, and selects the flavour of a drug compatibilities such as Semen Sojae Preparatum, the Radix Astragali, Radix Notoginseng, Fructus Ligustri Lucidi, through the optimization of the extracting method such as orthogonal experiment, and the active component (E) required for reaching in compatibe drug; By all to A, B, C, D, E effective site, particularly D component is assembled in soft capsule dosage form as proliposome, forms novel " proliposome assembling soft capsule " compound Chinese medicinal preparation.
Prescription is resolved: the vertical Ganoderma spore of we, mussel are monarch drug, the strengthening vital QI to eliminate pathogenic factors that both performances all have, anti-cancer and inhibiting tumor effect.Two taste principal agent compatibilities, a Wen Yihan, the property of medicine is complementary, and effect is remarkable.With Radix Ginseng, the Radix Astragali to medicine for ministerial drug, the gas of tonifying five ZANG-organs is apt to by Radix Ginseng, QI invigorating and hold concurrently can yin nourishing, keeps and do not walk; Radix Astragali sweet in the mouth is warm in nature, is apt to away flesh table, QI invigorating and hold concurrently can YANG invigorating, walks and does not keep.Two medicine mutual reinforcement between pairings one are walked one and are kept, and invigorating primordial QI, promoting generation of vital essence and blood, negative and positive are taken into account, and enhancing body battalion defends ability, significantly improves immune function of human body, assists a ruler in governing a country Ganoderma spore, mussel two taste monarch drug, invigorate vital energy and yang, YIN nourishing and the production of body fluid promoting, strengthens strengthening the body resistance and cancer inhibitting and killing cell effect.In side, Fructus Ligustri Lucidi, Radix Notoginseng are adjuvant drug, and Fructus Ligustri Lucidi is cool in nature, sweet in the mouth, hardship.Return liver, kidney channel, nourishing the liver and kidney, improving eyesight black hair.Fructus Ligustri Lucidi itself has active anticancer, more can assist a ruler in governing a country Radix Ginseng, the Radix Astragali to medicine, and body can be made greatly to improve tumour immunity function.Radix Notoginseng sweet in the mouth warm in nature, returns liver, stomach warp, and dissipating blood stasis stops blooding, subduing swelling and relieving pain.Radix Notoginseng has " adaptogen " sample dual regulation, can play according to cancer patient's individual instances and just help or using corrigent effect, assist a ruler in governing a country Fang Zhongjun, ministerial drug and play QI invigorating complement effect better, strengthen blood circulation and channel invigorating effect, have hemostasis and pain-relieving effect to the pain of carcinoma and bleeding.The cool complementation of two taste adjuvant drug property of medicine temperature, effect negative and positive are taken into account, and are conducive to the dynamic equilibrium of two-way adjustment cancer patient organismic internal environment.Compatibility Pericarpium Citri Reticulatae, Semen Sojae Preparatum are for making medicine, and Pericarpium Citri Reticulatae is regulated the flow of vital energy the knot that disappears, drying dampness to eliminate phlegm, help patient to eliminate the phlegm-damp cancer source of alluvial in body; Induce sweat relieving restlessness, a surname of Semen Sojae Preparatum sends out hot and suffocating, can remove patient and smoulder the mental pressure such as annoyance agitation, restlessness of asrhenia type and insomnia in the heart, while itself there is the effect of cancer inhibitting and killing tumor, effectively can coordinate monarch, ministerial drug, enhancing active anticancer.All medicines coordinate, and play strengthening vital QI to eliminate pathogenic factors , Xiao Disorder pain relieving altogether, cancer inhibitting and killing tumor, regulate equilibrium between yin and yang, overcome the effects such as ambition unhappiness.Play the tumor-inhibiting action of our multicomponent, multipath, Mutiple Targets thus, comprehensive transfer animal economy ability, reach the tidemark of anti-curing cancers, namely people-oriented, treating both the principal and secondary aspects of a disease; Cancer inhibitting and killing tumor is thoroughly cured best, and band tumor existence " peaceful coexistence " also can.
The prescription of " compound ganoderma spore mussel pro-liposome assembling soft capsule " provided by the invention preparation comprises the medical materials such as Ganoderma spore, Extract of Cyclina Sinensis, Radix Ginseng, the Radix Astragali, Fructus Ligustri Lucidi, Radix Notoginseng, Semen Sojae Preparatum, Pericarpium Citri Reticulatae, and various proportioning raw materials is by weight: Ganoderma spore 500g ~ 1000g, Extract of Cyclina Sinensis 50g ~ 200g, Radix Ginseng 200 ~ 350g, Radix Astragali 200g ~ 350g, Fructus Ligustri Lucidi 150g ~ 250g, Radix Notoginseng 100 ~ 200g, Semen Sojae Preparatum 200 ~ 350g, Pericarpium Citri Reticulatae 100 ~ 200g.
" compound ganoderma spore mussel pro-liposome assembling soft capsule " of the present invention preparation method, concrete steps are as follows:
(1) extraction and isolation of effective ingredient in Chinese
1) triterpene acids extraction and separation process in Ganoderma spore: supercritical fluid extraction: get Ganoderma spore powder with cellular wall broken, uses supercritical CO 2extraction, extractant condition is: extracting pressure 10 ~ 25Mpa, extraction temperature 40 ~ 60 DEG C, extraction time 1 ~ 2 hour, collects extraction of essential oil thing; The residue extracting quintessence oil adds the ethanol of 10 ~ 30% as entrainer extraction Ganoderma triterpenoids acids component, and extractant condition is: extracting pressure 20 ~ 40Mpa, extraction temperature 45 ~ 65 DEG C, extraction time 1.5 ~ 3.0 hours.
2) triterpene saponin extraction and separation process: by Radix Ginseng, the Radix Astragali, Fructus Ligustri Lucidi, Radix Notoginseng, extracts 2 ~ 3 times with 60 ~ 80% alcohol heating reflux of 5 ~ 8 times amount volumes, each 1 ~ 3 hour, concentrating under reduced pressure, reclaim ethanol, vacuum spray drying, pulverize, obtain extract fine powder, detect wherein triterpene saponin content, require to reach more than 50%, need if desired to carry out macroporous resin column chromatography polishing purification further, the medicinal residues extracted retain, and are made for next step Polyose extraction.
3) polysaccharide extraction and separation process: will the Radix Ginseng of saponin, the Radix Astragali, Fructus Ligustri Lucidi, the medicinal residues of Radix Notoginseng and above-mentioned through supercritical CO be extracted 2ganoderma spore medicinal residues after extraction, extract 2 ~ 3 times with 20 ~ 50% alcohol heating reflux of 4 ~ 6 times amount volumes, each 1 ~ 3 hour, concentrating under reduced pressure, vacuum drying, pulverizes, obtain polyoses extract fine powder, detect wherein polyoses content, require to reach more than 50%, need if desired to carry out membrance separation polishing purification further.
4) isoflavone and the enrichment of flavones extraction and isolation: Semen Sojae Preparatum, tangerine peel powder are broken into coarse powder, application supercritical CO 2extraction, extractant condition is: extracting pressure 15 ~ 30Mpa, extraction temperature 40 ~ 60 DEG C, extraction time 1 ~ 2 hour, collects extraction of essential oil thing; The residue extracting quintessence oil adds ethanol extraction separating isoflavone and the flavones of 70 ~ 90%, concentrating under reduced pressure, reclaim ethanol, vacuum spray drying, pulverize, obtain flavone and isoflavone extract fine powder, detect wherein total flavonoid content, requirement reaches more than 50%, needs if desired to carry out macroporous resin column chromatography polishing purification further.
(2) preparation of mussel glycoprotein (HBTDB) extraction separation and purification and pro-liposome
1) HBTDB extraction and separation process: by mussel meat, homogenate, by 3 ~ 6 times of less than 10 DEG C cold water extraction 1 ~ 2 time, centrifugation, supernatant is separated, with 30 ~ 90% ethanol precipitate with ethanol, precipitate 2 ~ 8 times of 6 ~ 16% sodium chloride solutions to precipitate quality are saltoutd, centrifugation, gets supernatant ultrafiltration, concentrated postlyophilization, obtain HBTDB, detect wherein glycoprotein Content, require to reach more than 50%, need if desired to use membrance separation polishing purification further.
2) preparation technology of HBTDB pro-liposome: with reverse phase evaporation, get HBTDB, liposome adjuvant, freeze drying protectant (mannitol: lecithin: cholesterol: HBTDB=200 ~ 400:100 ~ 200:60 ~ 150:30 ~ 100), the lecithin of recipe quantity and cholesterol are dissolved in ether, HBTDB and mannitol are dissolved in normal saline, by two kinds of solution mixing, sonic oscillation 1 ~ 10 minute, the rotary evaporation that reduces pressure at 30 DEG C ~ 50 DEG C removing ether, obtain HBTDB liposome suspendible emulsion, detecting envelop rate should reach more than 65%; Get qualified HBTDB liposome, put in freezer dryer and carry out drying by operating process, obtain HBTDB pro-liposome.
(3) pro-liposome assembling soft capsule preparation
By the medical material extraction of essential oil thing that obtains through supercritical fluid extraction in 40 ~ 50 DEG C of insulations, above-mentioned various effective part extract fine powder is mixed homogeneously with HBTDB pro-liposome, add in the quintessence oil of insulation, appropriate medicinal oil Suitable content object sum viscosity can be added if desired, repeatedly grind through colloid mill, abundant mix homogeneously, crosses 80 ~ 120 mesh sieves, repetitive operation if desired 2 ~ 4 times, by contents homogenised one-tenth suspension, be prepared into soft capsule, granulate, washing, dry, packaging, inspection, obtains qualified finished product.
The novelty that " compound ganoderma spore mussel pro-liposome assembling soft capsule " of the present invention has: 1) adopt supercritical CO 2fluid extraction new and high technology, fat-soluble for lucid ganoderma spore essence wet goods functional component is fully extracted and enrichment, as entrainer and under optimizing other conditions, the triterpenes such as Ganodenic acid is separated and enrichment adding ethanol, makes the yield of Ganoderma spore quintessence oil bring up to present more than 10% from about 4% of traditional conventional organic solvent extraction; The yield of Ganoderma triterpenoids effective kind part improves 2 times, and enriched concentration reaches more than 50%.This technology has cut off the disadvantage of traditional handicraft with a large amount of organic solvent, and this technique is environmentally friendly, safety non-toxic.2) product formulation adopts " pro-liposome assembling soft capsule " novel form, in Chinese medicine preparation, there is not yet report.Mussel glycoprotein (HBTDB) development is liposome by the present invention; add freeze drying protectant again; being prepared into pro-liposome after lyophilization is assembled in soft capsule; preparation process adopts colloid mill to homogenize technology; make extract powder micronization; can suspendible even with quintessence oil, solve solid phase and the easy layering of oil phase in soft capsule preparation process, not easily uniform shortcoming.The product of " pro-liposome assembling soft capsule " preparation that the present invention makes has anti cancer target administration advantage, and dissolution rate is high, improves bioavailability in body; Product design of the present invention is attractive in appearance, and active component obtains abundant enrichment and concentrates, therefore dosage is little, taking convenience, overcomes " black, large, thick " of current similar compound Chinese medicinal preparation and is difficult to the shortcoming taken.3) in this product prescription, various effective site combination has again new lifting on Chinese medicine combination principle basis, in side, various effective site plays multi-agent synergy effect, antitumor and immunomodulating drug effect are eliminating evil from strengthening the body resistance, tumor suppression, efficacy enhancing and toxicity reducing and immunoregulation four aspect Comprehensive consideration, are different from simple cell poison cancer therapy drug.4) experimentation proves that the combination of effective site in this product prescription can the telomerase of effectively direct or indirect anticancer, anticancer DNA holder can flutter the activity of isomerase (TOPO), promote DNA break, cancer cell specific induction of apoptosis; General Promotion body's immunity, promote lymphopoiesis, improve the phagocytic activity of macrophage, NK cell, T cell, remarkable enhancing non-specific immunity function and humoral immune function, reverse the immunosuppressant phenomenon caused because of cyclophosphamide, leukocyte increasing, Promote immunity globulin, enhancing antibody generate, increase NK cytoactive, thus kill and wound, engulf cancerous cell, body is stimulated to produce interferon-Y (INF-Y) and tumor necrosis factor alpha a (INF-a), cancer cell specific induction of apoptosis; The routine chemicotherapy patient of Clinical practice more than 30, obviously can alleviate toxic and side effects, can tolerate and adhere to the course for the treatment of, and display this product has efficacy enhancing and toxicity reducing effect to chemotherapy, radiotherapy.
Detailed description of the invention
According to the formula of " compound ganoderma spore mussel pro-liposome assembling soft capsule " of the present invention, select three groups of prescriptions: prescription one: Ganoderma spore 500g, Extract of Cyclina Sinensis 100g, Radix Ginseng 200g, Radix Astragali 200g, Fructus Ligustri Lucidi 150g, Radix Notoginseng 100g, Semen Sojae Preparatum 200g, Pericarpium Citri Reticulatae 100g.
Prescription two: Ganoderma spore 750g, Extract of Cyclina Sinensis 150g, Radix Ginseng 250g, Radix Astragali 250g, Fructus Ligustri Lucidi 200g, Radix Notoginseng 150g, Semen Sojae Preparatum 300g, Pericarpium Citri Reticulatae 150.
Prescription three: Ganoderma spore 1000g, Extract of Cyclina Sinensis 200g, Radix Ginseng 350g, Radix Astragali 350g, Fructus Ligustri Lucidi 250g, Radix Notoginseng 200g, Semen Sojae Preparatum 350g, Pericarpium Citri Reticulatae 200g.
The extraction separation method example 1 of the effective ingredient in Chinese group of compound ganoderma spore mussel pro-liposome assembling soft capsule:
1) in Ganoderma spore, triterpene acids supercritical fluid extraction process conditions are: extracting pressure 15Mpa, extraction temperature 40 DEG C, and extraction time 1 hour, obtains Ganoderma spore quintessence oil; The residue extracting quintessence oil adds the ethanol of 10% as entrainer extraction ganoderma lucidum triterpene component, and extractant condition is: extracting pressure 25Mpa, extraction temperature 50 DEG C, extraction time 1.5 hours.
2) triterpene saponin extraction and separation process condition is: Radix Ginseng, the Radix Astragali, Fructus Ligustri Lucidi, Radix Notoginseng powder are broken into coarse powder, 2 times are extracted with 60% alcohol heating reflux of 5 times amount volumes, each 1.5 hours, concentrating under reduced pressure, reclaim ethanol, vacuum spray drying, pulverizes, obtains total triterpene saponins extract.
3) polysaccharide extraction and separation process condition is: will extract the Radix Ginseng of saponin, the Radix Astragali, Fructus Ligustri Lucidi, the medicinal residues of Radix Notoginseng and above-mentioned through supercritical CO 2ganoderma spore medicinal residues after extraction, extract 2 times with 20% alcohol heating reflux of 4 times amount volumes, each 1.5 hours, and concentrating under reduced pressure, vacuum drying, obtains total polysaccharide extractive.
4) total flavonoid extraction and isolation: Semen Sojae Preparatum, tangerine peel powder are broken into coarse powder, first through supercritical CO 2quintessence oil in oils and fats and Pericarpium Citri Reticulatae in extract and separate Semen Sojae Preparatum, extractant condition is: extracting pressure 15Mpa, extraction temperature 40 DEG C, extraction time 1 hour, collects extraction of essential oil thing, residue adds ethanol extraction separating isoflavone and the flavones of 65%, concentrating under reduced pressure, reclaims ethanol, vacuum spray drying, pulverize, obtain total flavonoid extract.
The extraction separation method example 2 of the effective ingredient in Chinese group of compound ganoderma spore mussel pro-liposome assembling soft capsule:
1) in Ganoderma spore, triterpene acids supercritical fluid extraction process conditions are: extracting pressure 25Mpa, extraction temperature 50 DEG C, and extraction time 2 hours, obtains Ganoderma spore quintessence oil; The residue extracting quintessence oil adds the ethanol of 15% as entrainer extraction ganoderma lucidum triterpene component, and extractant condition is: extracting pressure 30Mpa, extraction temperature 55 DEG C, extraction time 2 hours.
2) triterpene saponin extraction and separation process condition is: Radix Ginseng, the Radix Astragali, Fructus Ligustri Lucidi, Radix Notoginseng powder are broken into coarse powder, extracts 2 times with 70% alcohol heating reflux of 6 times amount volumes, each 2 hours, concentrating under reduced pressure, reclaims ethanol, vacuum spray drying, pulverize, obtain total triterpene saponins extract.
3) polysaccharide extraction and separation process condition is: will extract the Radix Ginseng of saponin, the Radix Astragali, Fructus Ligustri Lucidi, the medicinal residues of Radix Notoginseng and above-mentioned through supercritical CO 2ganoderma spore medicinal residues after extraction, extract 2 times with 40% alcohol heating reflux of 5 times amount volumes, each 2 hours, and concentrating under reduced pressure, vacuum drying, obtains total polysaccharide extractive.
4) total flavonoid extraction and isolation: Semen Sojae Preparatum, tangerine peel powder are broken into coarse powder, first through supercritical CO 2quintessence oil in oils and fats and Pericarpium Citri Reticulatae in extract and separate Semen Sojae Preparatum, extractant condition is: extracting pressure 20Mpa, extraction temperature 50 DEG C, extraction time 2 hours, collects extraction of essential oil thing, residue adds ethanol extraction separating isoflavone and the flavones of 80%, concentrating under reduced pressure, reclaims ethanol, vacuum spray drying, pulverize, obtain total flavonoid extract.
The extraction separation method example 3 of the effective ingredient in Chinese group of compound ganoderma spore mussel pro-liposome assembling soft capsule:
1) in Ganoderma spore, triterpene acids supercritical fluid extraction process conditions are: extracting pressure 30Mpa, extraction temperature 55 DEG C, and extraction time 1 hour, obtains Ganoderma spore quintessence oil; The residue extracting quintessence oil adds the ethanol of 20% as entrainer extraction ganoderma lucidum triterpene component, and extractant condition is: extracting pressure 35Mpa, extraction temperature 60 DEG C, extraction time 2.5 hours.
2) triterpene saponin extraction and separation process condition is: Radix Ginseng, the Radix Astragali, Fructus Ligustri Lucidi, Radix Notoginseng powder are broken into coarse powder, 3 times are extracted with 80% alcohol heating reflux of 8 times amount volumes, each 1.5 hours, concentrating under reduced pressure, reclaim ethanol, vacuum spray drying, pulverizes, obtains total triterpene saponins extract.
3) polysaccharide extraction and separation process condition is: will extract the Radix Ginseng of saponin, the Radix Astragali, Fructus Ligustri Lucidi, the medicinal residues of Radix Notoginseng and above-mentioned through supercritical CO 2ganoderma spore medicinal residues after extraction, extract 2 times with 50% alcohol heating reflux of 6 times amount volumes, each 2.5 hours, and concentrating under reduced pressure, vacuum drying, obtains total polysaccharide extractive.
4) total flavonoid extraction and isolation: Semen Sojae Preparatum, tangerine peel powder are broken into coarse powder, first through supercritical CO 2quintessence oil in oils and fats and Pericarpium Citri Reticulatae in extract and separate Semen Sojae Preparatum, extractant condition is: extracting pressure 25Mpa, extraction temperature 55 DEG C, extraction time 2.5 hours, collects extraction of essential oil thing, residue adds ethanol extraction separating isoflavone and the flavones of 80%, concentrating under reduced pressure, reclaims ethanol, vacuum spray drying, pulverize, obtain total flavonoid extract.
According to the extraction separation method of the effective ingredient in Chinese group of compound ganoderma spore mussel pro-liposome assembling soft capsule, preparation technology's example 1 of mussel glycoprotein (HBTDB) extraction separation and purification and pro-liposome:
HBTDB extraction and separation process is: by mussel meat, homogenate, with 3 times of less than 10 DEG C cold water extraction 1 time, centrifugation, supernatant is separated, with 45% ethanol precipitate with ethanol, precipitate 4 times of 6% sodium chloride solutions to precipitate quality are saltoutd, and centrifugation, gets supernatant ultrafiltration, concentrated postlyophilization, obtains HBTDB.The preparation technology of HBTDB pro-liposome: with reverse phase evaporation; get HBTDB, liposome adjuvant, freeze drying protectant (mannitol: lecithin: cholesterol: HBTDB=200:100:60:30); the lecithin of recipe quantity and cholesterol are dissolved in ether; HBTDB and mannitol are dissolved in normal saline; by two kinds of solution mixing; sonic oscillation 1 minute; the rotary evaporation that reduces pressure at 45 DEG C removing ether; obtain HBTDB liposome suspendible emulsion; put in freezer dryer and carry out drying by operating process, obtain HBTDB pro-liposome.
According to the extraction separation method of the effective ingredient in Chinese group of compound ganoderma spore mussel pro-liposome assembling soft capsule, preparation technology's example 2 of mussel glycoprotein (HBTDB) extraction separation and purification and pro-liposome:
HBTDB extraction and separation process is: by mussel meat, homogenate, with 4 times of less than 10 DEG C cold water extraction 2 times, centrifugation, supernatant is separated, with 55% ethanol precipitate with ethanol, precipitate 5 times of 8% sodium chloride solutions to precipitate quality are saltoutd, and centrifugation, gets supernatant ultrafiltration, concentrated postlyophilization, obtains HBTDB.The preparation technology of HBTDB pro-liposome: with reverse phase evaporation; get HBTDB, liposome adjuvant, freeze drying protectant (mannitol: lecithin: cholesterol: HBTDB=300: 200: 80:40); the lecithin of recipe quantity and cholesterol are dissolved in ether; HBTDB and mannitol are dissolved in normal saline; by two kinds of solution mixing; sonic oscillation 2 minutes; the rotary evaporation that reduces pressure at 50 DEG C removing ether; obtain HBTDB liposome suspendible emulsion; put in freezer dryer and carry out drying by operating process, obtain HBTDB pro-liposome.
According to the extraction separation method of the effective ingredient in Chinese group of compound ganoderma spore mussel pro-liposome assembling soft capsule, preparation technology's example 3 of mussel glycoprotein (HBTDB) extraction separation and purification and pro-liposome:
HBTDB extraction and separation process is: by mussel meat, homogenate, with 6 times of less than 10 DEG C cold water extraction 2 times, centrifugation, supernatant is separated, with 65% ethanol precipitate with ethanol, precipitate 6 times of 10% sodium chloride solutions to precipitate quality are saltoutd, and centrifugation, gets supernatant ultrafiltration, concentrated postlyophilization, obtains HBTDB.The preparation technology of HBTDB pro-liposome: with reverse phase evaporation; get HBTDB, liposome adjuvant, freeze drying protectant (mannitol: lecithin: cholesterol: HBTDB=350:250:100:50); the lecithin of recipe quantity and cholesterol are dissolved in ether; HBTDB and mannitol are dissolved in normal saline; by two kinds of solution mixing; sonic oscillation 3 minutes; the rotary evaporation that reduces pressure at 60 DEG C removing ether; obtain HBTDB liposome suspendible emulsion; put in freezer dryer and carry out drying by operating process, obtain HBTDB pro-liposome.
According to the formulation preparation method of compound ganoderma spore mussel pro-liposome assembling soft capsule, concrete steps are: by the medical material extraction of essential oil thing mixing obtained through supercritical fluid extraction, 50 DEG C of insulations, above-mentioned various effective part extract fine powder is mixed homogeneously with HBTDB pro-liposome, add in the quintessence oil of insulation, appropriate medicinal oil Suitable content object sum viscosity can be added if desired, repeatedly grind through colloid mill, abundant mix homogeneously, cross 120 mesh sieves, by contents homogenised one-tenth suspension, be prepared into soft capsule, place, granulate, washing, dry, packaging, inspection, obtain qualified finished product.
Prepare gained " compound ganoderma spore mussel pro-liposome assembling soft capsule " as stated above, its quality index is in table 1 after testing:
Table 1. effective ingredient index
The antitumor drug effect result of the test of compound ganoderma spore mussel pro-liposome assembling soft capsule of the present invention.
(1) mouse tumor model: mice intestinal cancer C26, mice Lewi pulmonary carcinoma.To be gone down to posterity maintenance by pharmacological room of Shanghai Institute of Pharmaceutical Industry in tumor source.C57BL/6 mice is provided by Shanghai City Chinese Academy of Sciences Experimental Animal Center, the quality certification number: SCXK2013-0003.Animal occupancy permit number: SYXK (Shanghai) 2004-0015.Animal male and female all can, often criticize experiment and use same sex, often organize 10 mices.Dosage is arranged: compound ganoderma spore mussel pro-liposome assembling soft capsule content 400mg/kg/d, 200mg/kg/d and 100mg/kg/d.Gastric infusion, is non-cytotoxicity class because of this product therefore starts administration, administration every day 1 time, continuous 10 days in 24 hours after tumor inoculation.Experimental control: negative control group gives blank liposome, the same test group of dosage regimen.Positive controls cisplatin for inj DDP7mg/kg, intraperitoneal administration 2 times, the 1st, 3 day.Put to death each treated animal after 2 weeks, dissect, get tumor and weigh, calculate tumor control rate.Result: result of the test shows, the tumour inhibiting rate of this product to high, medium and low three the dosage groups of mice intestinal cancer C26 is respectively 65.36%, 53.29%, 48.71%; 61.96%, 50.77%, 42.69% is respectively to the tumour inhibiting rate of high, medium and low three the dosage groups of mice Lewi pulmonary carcinoma, shows certain dose-effect relationship.
(2) human tumor nude mouse xenograft model: human hepatoma QGY, Human Gastric Cancer MNK-45.Design is the same with operation.Result of the test shows: this product is respectively 65.36%, 53.29%, 48.71% to human hepatoma QGY xenotransplantation in the tumour inhibiting rate of high, medium and low three the dosage groups of immunodeficiency nude mouse model; In the tumour inhibiting rate of high, medium and low three the dosage groups of immunodeficiency nude mouse model, 35.45%, 49.38%, 64.16% is respectively to Human Gastric Cancer MNK-45 xenotransplantation.
(3) the enhancing immunologic function result of the test of " compound ganoderma spore mussel pro-liposome assembling soft capsule " of the present invention.
Per os to give 400mg/kg/d, 200mg/kg/d, 100mg/kg/d (20 times of the per kg body weight per day recommended intake that is equivalent to respectively to be grown up every day, 10 times, 5 times) compound ganoderma spore mussel pro-liposome assembling soft capsule content 30 days, every result of the test is as follows:
1) on the impact of the weight of animals before and after test: after experiment, each animal subject dosage group mice increases weight in solvent control group without the significance difference opposite sex (P > 0.05).
2) on the impact of the ratio of internal organs body weight: with solvent control group ratio, each group mouse thymus body ratio, spleen body ratio no difference of science of statistics (P > 0.05).
3) cellular immune function test: the mice spleen of induction is to target cell conversion test (mtt assay): each dosage group optical density difference is all higher than solvent control group, and increase with dosage and increase, middle and high dosage group has statistical significant difference (P < 0.05, P < 0.01).Point out middle and high dosage this product to have and strengthen mouse lymphocyte multiplication capacity.The elimination Tardive allergy (DTH) that dinitrofluorobenzene (DNFB) is induced: the auricle swelling degree of each dosage group mice is all higher than solvent control class value, and increase with dosage and increase, each dosage group all has statistically-significant difference (P < 0.05, P < 0.01).Prompting this product can strengthen the mice Tardive allergy that DNFB causes.According to above two result of the tests, show that compound ganoderma spore mussel pro-liposome assembling soft capsule has and strengthen mouse cell immunologic function.
4) humoral immune function test: antibody-producting cell detects (Jeme improves ripple electrical method): each dosage group plaque number is all higher than solvent control group, middle and high dosage group plaque has significant difference (P < 0.05, P < 0.01) compared with matched group.Middle and high dosage this product is pointed out to have the ability strengthening mouse antibodies cellulation.Serolysin test: each dosage group mice serum half hemolysis value (HC 50) all higher than contrast class value, high dose group has remarkable significant difference (P < 0.05).Prompting high dose this product can strengthen the ability that mice produces serum hemolysin.
5) monokaryon one macrophage function test: mice carbonic clearance is tested: the phagocytic index of each dosage group, compared with solvent control group, all has statistical significant difference (P < 0.05).Prompting this product has the effect obviously strengthening mice carbonic clearance ability.Turnover of Mouse Peritoneal Macrophages engulfs chicken red blood cell test (dripping sheet method): the monokaryon one megalith phagocytic percentage of each dosage group mice and phagocytic index are all higher than solvent control group analog value, middle and high dosage group phagocytic rate and each dosage group phagocytic index all have statistical discrepancy (P < 0.05, P < 0.01).Under pointing out this experimental condition, this product has the effect strengthening mouse peritoneal phagocyte phagocytic activity.
6) NK cytoactive detection: the NK cytoactive of each dosage group all higher than solvent control group, and has obvious significant difference (P < 0.05), shows that this product has the effect strengthening NK cells in mice activity.
According to enhancing immunity functional evaluation standard, show that compound ganoderma spore mussel pro-liposome assembling soft capsule has the function of enhancing immunity.
The toxicologic study result of compound ganoderma spore mussel pro-liposome assembling soft capsule of the present invention:
1) acute toxicity test in mice: this product LD 50for 11.56g/kg (with this product content gauge), be equivalent to 241.6 times of referrer's clinical dosage.
2) rat acute toxicity test: this product LD 50for 18.44g/kg (with this product content gauge), be equivalent to 384.1 times of referrer's clinical dosage.
3) rat takes this product 13 weeks toxicity tests continuously: this test is evaluate the continuous per os of this product to give the rat toxicity of 90 days and drug withdrawal 30 days convalescent reaction reversibilitys.Basic, normal, high dosage component Wei 1g/kg/d, 2g/kg/d, 3g/kg/d, experimental period every treated animal 20, female, male each 10, convalescent period animal per group 10, female, male each 5.Result of the test: the appetite of high dose group male rat reduces after 6 weeks in administration, movable few, recover normal after drug withdrawal.The instant animal of administration has no obvious toxic reaction, does not find death.Blood biochemical analysis checks without exception, at the end of take the weight of animals, two kinds of sexes all present the trend that body weight increases, there was no significant difference compared with blank group.Experiment terminates rear each group of every zootomy, and get liver, kidney, Stomach duodenum, spleen, testis, ovarian sections have no obvious substantive pathological change in basis of microscopic observation, administration group is similar with matched group situation.Show that each organ of examined rat does not all occur assembling the obviously relevant tissue pathologies change of soft capsule with compound ganoderma spore mussel pro-liposome.

Claims (8)

1. resist carcinoma effective novel synergistic herbal effective part group compound preparation-" compound ganoderma spore mussel pro-liposome assembling soft capsule " for one kind, it is characterized in that: formula comprises the medical material prescriptions such as Ganoderma spore, Extract of Cyclina Sinensis, Radix Ginseng, the Radix Astragali, Fructus Ligustri Lucidi, Radix Notoginseng, Semen Sojae Preparatum, Pericarpium Citri Reticulatae and is prepared from.Wherein, the combination of Ganoderma spore, Extract of Cyclina Sinensis, as the key agents of this product prescription, plays and strengthens the characteristic place that the effect of resisting carcinoma activity is this prescription.This product prepares 1000 soft capsule needed raw material medical material proportionings:
Ganoderma spore 500g ~ 1000g, Extract of Cyclina Sinensis 50g ~ 200g, Radix Ginseng 200 ~ 350g, Radix Astragali 200g ~ 350g, Fructus Ligustri Lucidi 150g ~ 250g, Radix Notoginseng 100 ~ 200g, Semen Sojae Preparatum 200 ~ 350g, Pericarpium Citri Reticulatae 100 ~ 200g.
2. " compound ganoderma spore mussel pro-liposome assembling soft capsule " according to claim 1, it is characterized in that: each raw medicinal material proportioning is:
Ganoderma spore 500g, Extract of Cyclina Sinensis 100g, Radix Ginseng 200g, Radix Astragali 200g, Fructus Ligustri Lucidi 150g, Radix Notoginseng 100g, Semen Sojae Preparatum 200g, Pericarpium Citri Reticulatae 100g.
3. " compound ganoderma spore mussel pro-liposome assembling soft capsule " according to claim 1, it is characterized in that: each raw medicinal material proportioning is:
Ganoderma spore 750g, Extract of Cyclina Sinensis 150g, Radix Ginseng 250g, Radix Astragali 250g, Fructus Ligustri Lucidi 200g, Radix Notoginseng 150g, Semen Sojae Preparatum 300g, Pericarpium Citri Reticulatae 150.
4. " compound ganoderma spore mussel pro-liposome assembling soft capsule " according to claim 1, it is characterized in that: each raw medicinal material proportioning is:
Ganoderma spore 1000g, Extract of Cyclina Sinensis 200g, Radix Ginseng 350g, Radix Astragali 350g, Fructus Ligustri Lucidi 250g, Radix Notoginseng 200g, Semen Sojae Preparatum 350g, Pericarpium Citri Reticulatae 200g.
5. " compound ganoderma spore mussel pro-liposome assembling soft capsule " according to claims 1 to 4, is characterized in that in every capsules, active component content's index is:
Total triterpene acid >=100mg/ grain, mussel glycoprotein (HBTDB) >=80mg/ grain, total polysaccharides >=100mg/ grain, genistein >=20mg/ grain.
6. the extraction separation method of the effective ingredient in Chinese group of " compound ganoderma spore mussel pro-liposome assembling soft capsule " according to claims 1 to 5, is characterized in that concrete steps are:
1) triterpene acids extraction and separation process in Ganoderma spore: supercritical fluid extraction: get Ganoderma spore powder with cellular wall broken, uses supercritical CO 2extraction, extractant condition is: extracting pressure 10 ~ 25Mpa, extraction temperature 40 ~ 60 DEG C, extraction time 1 ~ 2 hour, collects extraction of essential oil thing; The residue extracting quintessence oil adds the ethanol of 5 ~ 20% as entrainer extraction ganoderma lucidum triterpene component, and extractant condition is: extracting pressure 20 ~ 40Mpa, extraction temperature 45 ~ 65 DEG C, extraction time 1.5 ~ 3.0 hours;
2) triterpene saponin extraction and separation process: by Radix Ginseng, the Radix Astragali, Fructus Ligustri Lucidi, Radix Notoginseng, extracts 2 ~ 3 times with 60 ~ 80% alcohol heating reflux of 5 ~ 8 times amount volumes, each 1 ~ 3 hour, concentrating under reduced pressure, reclaim ethanol, vacuum spray drying, pulverize, obtain extract fine powder, detect wherein triterpene saponin content, require to reach more than 50%, need if desired to carry out macroporous resin column chromatography polishing purification further, the medicinal residues extracted retain, and are made for next step Polyose extraction;
3) polysaccharide extraction and separation process: will the Radix Ginseng of saponin, the Radix Astragali, Fructus Ligustri Lucidi, the medicinal residues of Radix Notoginseng and above-mentioned through supercritical CO be extracted 2ganoderma spore medicinal residues after extraction, extract 2 ~ 3 times with 20 ~ 50% alcohol heating reflux of 4 ~ 6 times amount volumes, each 1 ~ 3 hour, concentrating under reduced pressure, vacuum drying, pulverizes, obtain polyoses extract fine powder, detect wherein polyoses content, require to reach more than 50%, need if desired to carry out membrance separation polishing purification further;
4) isoflavone and the enrichment of flavones extraction and isolation: Semen Sojae Preparatum, tangerine peel powder are broken into coarse powder, application supercritical CO 2extraction, extractant condition is: extracting pressure 15 ~ 30Mpa, extraction temperature 40 ~ 60 DEG C, extraction time 1 ~ 2 hour, collects extraction of essential oil thing; The residue extracting quintessence oil adds ethanol extraction separating isoflavone and the flavones of 65 ~ 90%, concentrating under reduced pressure, reclaim ethanol, vacuum spray drying, pulverize, obtain flavone and isoflavone extract fine powder, detect wherein total flavonoid content, requirement reaches more than 50%, needs if desired to carry out silica gel column chromatography refinement purification further.
7. the extraction separation method of the effective ingredient in Chinese group of " compound ganoderma spore mussel pro-liposome assembling soft capsule " according to claims 1 to 6, the preparation of mussel glycoprotein (HBTDB) extraction separation and purification and pro-liposome, is characterized in that concrete steps are:
1) HBTDB extraction and separation process: by mussel meat, homogenate, by 3 ~ 6 times of less than 10 DEG C cold water extraction 1 ~ 2 time, centrifugation, supernatant is separated, with 30 ~ 90% ethanol precipitate with ethanol, precipitate 2 ~ 8 times of 6 ~ 16% sodium chloride solutions to precipitate quality are saltoutd, centrifugation, gets supernatant ultrafiltration, concentrated postlyophilization, obtain HBTDB, detect wherein glycoprotein Content, require to reach more than 50%, need if desired to use membrance separation polishing purification further;
2) preparation technology of HBTDB pro-liposome: with reverse phase evaporation, get HBTDB, liposome adjuvant, freeze drying protectant (mannitol: lecithin: cholesterol: HBTDB=200 ~ 400:100 ~ 200:60 ~ 150:30 ~ 100), the lecithin of recipe quantity and cholesterol are dissolved in ether, HBTDB and mannitol are dissolved in normal saline, by two kinds of solution mixing, sonic oscillation 1 ~ 10 minute, the rotary evaporation that reduces pressure at 30 DEG C ~ 50 DEG C removing ether, obtain HBTDB liposome suspendible emulsion, detecting envelop rate should reach more than 65%; Get qualified HBTDB liposome, put in freezer dryer and carry out drying by operating process, obtain HBTDB pro-liposome.
8. the finished product preparation preparation method of " compound ganoderma spore mussel pro-liposome assembling soft capsule " according to claims 1 to 7, is characterized in that concrete steps are:
By the medical material extraction of essential oil thing mixing obtained through supercritical fluid extraction, 40 ~ 50 DEG C of insulations, above-mentioned various effective part extract fine powder is mixed homogeneously with HBTDB pro-liposome, add in the quintessence oil of insulation, appropriate medicinal oil Suitable content object sum viscosity can be added if desired, repeatedly grind through colloid mill, abundant mix homogeneously, cross 80 ~ 120 mesh sieves, repetitive operation if desired 2 ~ 4 times, by contents homogenised one-tenth suspension, be prepared into soft capsule, place, granulate, washing, dry, packaging, inspection, obtains qualified finished product.
CN201310366989.3A 2013-08-21 2013-08-21 Compound formula for preparing soft capsule by pro-liposomes of ganoderma spore and freshwater mussel and preparation method thereof Pending CN104415132A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201310366989.3A CN104415132A (en) 2013-08-21 2013-08-21 Compound formula for preparing soft capsule by pro-liposomes of ganoderma spore and freshwater mussel and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201310366989.3A CN104415132A (en) 2013-08-21 2013-08-21 Compound formula for preparing soft capsule by pro-liposomes of ganoderma spore and freshwater mussel and preparation method thereof

Publications (1)

Publication Number Publication Date
CN104415132A true CN104415132A (en) 2015-03-18

Family

ID=52965955

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201310366989.3A Pending CN104415132A (en) 2013-08-21 2013-08-21 Compound formula for preparing soft capsule by pro-liposomes of ganoderma spore and freshwater mussel and preparation method thereof

Country Status (1)

Country Link
CN (1) CN104415132A (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106581443A (en) * 2016-10-12 2017-04-26 中国人民解放军第二军医大学 Traditional Chinese medicinal composition for preventing or treating military stress disorders and preparation method of traditional Chinese medicinal composition
CN109452622A (en) * 2018-11-15 2019-03-12 江苏康缘药业股份有限公司 A kind of compound ganoderma conidia powder and preparation method thereof

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1403147A (en) * 2002-10-24 2003-03-19 河北以岭医药研究院有限公司 Medicines composition for raising body's immunity and its prepn
CN101804165A (en) * 2010-04-16 2010-08-18 武汉枫叶红生物科技有限公司 Preparation for clearing nephrotoxicity, strengthening internal organs, enhancing physique and preventing aging and preparation method thereof
CN101816739A (en) * 2010-05-18 2010-09-01 昆明中药厂有限公司 Chinese medicinal composition and preparation method thereof
CN102228595A (en) * 2011-06-29 2011-11-02 闫来生 Chinese prescription energy pure Chinese medicine extract with anti-aging function
CN102277250A (en) * 2011-07-14 2011-12-14 葛双铭 Nourishing wine
CN102423438A (en) * 2011-12-27 2012-04-25 峨眉山天梁星制药有限公司 Anticancer double-prescription medicament and preparation method thereof

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1403147A (en) * 2002-10-24 2003-03-19 河北以岭医药研究院有限公司 Medicines composition for raising body's immunity and its prepn
CN101804165A (en) * 2010-04-16 2010-08-18 武汉枫叶红生物科技有限公司 Preparation for clearing nephrotoxicity, strengthening internal organs, enhancing physique and preventing aging and preparation method thereof
CN101816739A (en) * 2010-05-18 2010-09-01 昆明中药厂有限公司 Chinese medicinal composition and preparation method thereof
CN102228595A (en) * 2011-06-29 2011-11-02 闫来生 Chinese prescription energy pure Chinese medicine extract with anti-aging function
CN102277250A (en) * 2011-07-14 2011-12-14 葛双铭 Nourishing wine
CN102423438A (en) * 2011-12-27 2012-04-25 峨眉山天梁星制药有限公司 Anticancer double-prescription medicament and preparation method thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN106581443A (en) * 2016-10-12 2017-04-26 中国人民解放军第二军医大学 Traditional Chinese medicinal composition for preventing or treating military stress disorders and preparation method of traditional Chinese medicinal composition
CN106581443B (en) * 2016-10-12 2020-04-07 中国人民解放军第二军医大学 Traditional Chinese medicine composition for preventing or treating military stress disorder and preparation method thereof
CN109452622A (en) * 2018-11-15 2019-03-12 江苏康缘药业股份有限公司 A kind of compound ganoderma conidia powder and preparation method thereof

Similar Documents

Publication Publication Date Title
CN102178796A (en) Application of effective component of medulla Junci in preparing medicine, health food or cosmetic for resisting tumor or inhibiting angiogenesis
CN103621869B (en) Giant salamander oligosaccharide peptide health-care product
CN105943615A (en) Application of whole hemp extract in preparation of pancreatic cancer treating drug
CN1307131C (en) Antineoplastic extract of Chinese fan palm and its preparation method and uses
CN102114102B (en) Traditional Chinese medicine extract mixed preparation and application thereof
CN108524814A (en) A kind of Chinese medicine composition and preparation method thereof for reducing blood glucose
CN102114099B (en) Traditional Chinese medicine preparation for inhibiting liver cancer and preparation method thereof
CN101804103B (en) Drug for treating leukopenia caused by concurrent chemoradiotherapy of malignant tumor and preparation method thereof
CN101711848B (en) Chinese medicinal composition capable of adjunctively treating tumour
CN102247479B (en) Antitumor strong medicine and preparation method thereof
CN101537036A (en) Soap pod saponin extract as well as preparation method and application thereof
CN104415132A (en) Compound formula for preparing soft capsule by pro-liposomes of ganoderma spore and freshwater mussel and preparation method thereof
CN101732377B (en) Method for preparing anti-cancer total saponin extracts of oplopanax elatus nakai leaves
CN103385931B (en) Blood-sugar-lowering medicine composition
CN101062374B (en) Method for preparing Chinese traditional combination capsule for treating cancer and the product thereof
CN105250440B (en) A kind of lenitive Chinese medicine composition and its application
CN104127684B (en) Traditional Chinese medicine compound preparation for treating cancers
CN103142934A (en) Traditional Chinese medicinal composition for treating lung cancer and liver cancer
CN103169844B (en) Traditional Chinese medicine composition having anti-lung cancer and anti-liver cancer effects
CN103933472B (en) A kind of Traditional Chinese medicinal composition for treating lung cancer and liver cancer
CN100411663C (en) Anticancer compound Chinese medicine powder and its preparing method
CN102114065B (en) Longhairy antenoron herb and common threewingnut root Chinese medicinal composition with effect of preventing and treating cancer and application thereof
CN100464773C (en) Yixinshu Injecta for retaliation of Qi and pulse and promotion of blood circulation and removing of blood stasis, and preparing method therefor
CN104707101A (en) Liver cancer and lung cancer resistant traditional Chinese medicine composition and application thereof
CN104127545B (en) New application of murraya tetramera huang and extract thereof in preparation of medicines

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
WD01 Invention patent application deemed withdrawn after publication
WD01 Invention patent application deemed withdrawn after publication

Application publication date: 20150318