CN107898903A - A kind of lipidosome cream of anti-herpesvirus and preparation method thereof - Google Patents

A kind of lipidosome cream of anti-herpesvirus and preparation method thereof Download PDF

Info

Publication number
CN107898903A
CN107898903A CN201710948443.7A CN201710948443A CN107898903A CN 107898903 A CN107898903 A CN 107898903A CN 201710948443 A CN201710948443 A CN 201710948443A CN 107898903 A CN107898903 A CN 107898903A
Authority
CN
China
Prior art keywords
fruit
liposome
cherokee rose
aloe
water
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
CN201710948443.7A
Other languages
Chinese (zh)
Inventor
王妍
张英
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.)
Shenzhen Polytechnic
Original Assignee
Shenzhen Polytechnic
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 Shenzhen Polytechnic filed Critical Shenzhen Polytechnic
Priority to CN201710948443.7A priority Critical patent/CN107898903A/en
Publication of CN107898903A publication Critical patent/CN107898903A/en
Pending legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/185—Magnoliopsida (dicotyledons)
    • A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
    • A61K36/738—Rosa (rose)
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
    • A61K36/18—Magnoliophyta (angiosperms)
    • A61K36/88—Liliopsida (monocotyledons)
    • A61K36/886—Aloeaceae (Aloe family), e.g. aloe vera
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/22—Heterocyclic compounds, e.g. ascorbic acid, tocopherol or pyrrolidones
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/06—Ointments; Bases therefor; Other semi-solid forms, e.g. creams, sticks, gels
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/127—Liposomes
    • A61K9/1277—Processes for preparing; Proliposomes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation 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
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/331—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using water, e.g. cold water, infusion, tea, steam distillation, decoction
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation 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
    • A61K2236/30—Extraction of the material
    • A61K2236/33—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones
    • A61K2236/333—Extraction of the material involving extraction with hydrophilic solvents, e.g. lower alcohols, esters or ketones using mixed solvents, e.g. 70% EtOH
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation 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
    • A61K2236/30—Extraction of the material
    • A61K2236/39—Complex extraction schemes, e.g. fractionation or repeated extraction steps
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K2236/00—Isolation 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
    • A61K2236/50—Methods involving additional extraction steps
    • A61K2236/53—Liquid-solid separation, e.g. centrifugation, sedimentation or crystallization

Landscapes

  • Health & Medical Sciences (AREA)
  • Natural Medicines & Medicinal Plants (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Epidemiology (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Engineering & Computer Science (AREA)
  • Mycology (AREA)
  • Alternative & Traditional Medicine (AREA)
  • Biotechnology (AREA)
  • Botany (AREA)
  • Medical Informatics (AREA)
  • Microbiology (AREA)
  • Dispersion Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Medicines Containing Plant Substances (AREA)
  • Cosmetics (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)

Abstract

The present invention provides a kind of lipidosome cream of anti-herpesvirus and preparation method thereof.The lipidosome cream forms liposome using the active ingredient in aloe polysaccharide and the fruit of Cherokee rose as active ingredient, using preferable liposome accessory package by aloe polysaccharide and fruit of Cherokee rose general flavone, lifts medicine stability and release rate；The percutaneous permeability of medicine is improved using preferable emulsifiable paste auxiliary material.The lipidosome cream has good effect of drugs, and without obvious toxic-side effects, has considerable application value.

Description

A kind of lipidosome cream of anti-herpesvirus and preparation method thereof
Technical field
The invention belongs to formulation art, and in particular to a kind of emulsifiable paste of anti-blister sore and preparation method thereof.
Background technology
Aloe (Aloe vera var chinesis (haw.) Berg.) contains abundant natural VE, C, A and B race Vitamin：Mineral element (such as potassium, zinc, selenium)；Amino acid etc., these be all to maintain the body youth often basic nutrition element. In addition, beauty and skin care and effective component as specific to aloe polysaccharide, phenolic compound etc. are all aloe.These special compositions and Basic beautifying nourishing element is together, common to complete the good beautifying skin-protection function of aloe and other effects.The polysaccharide contained in aloe It is the material with biological effect, and recent year studies its extraction process, studies the popular domain of its biological effect.
Aloe polysaccharide can adjust the cellular immunity and humoral immunity level of body, activate the Lang Hanxi of skin base layer Born of the same parents, strengthen its immune function and repair function in local skin, promote its remove skin pigment, inoxidizability infringement (including Ultraviolet), elasticity etc. of anti-aging increase skin.
Also contain aloe-emodin and barbaloin in aloe, this two classes composition can promote to wriggle inside large intestine and big intestinal juice Secretion, increase fat spin enzymatic activity, have large intestine laxative, orectic effect.Aloe promotes the excretion of interior metabolism product, subtracts Lack its residence time in vivo, improve organismic internal environment and skin metabolism, make skin full of vigour.
Contain many lipid oxidation enzyme compounds (such as flavonoids) that can suppress Antioxidation in aloe.Can be with Prevention of arterial atherosis, improves body, the supply of blood flow of skin and microcirculation, and the nutrition supply of abundance is provided for skin, is increased The vigor of strong skin epithelial cell metabolism.Contain steroid and steroids compound, steroid and steroids compound in aloe And some very important microcomponents that people retain youthful vitality, they are thanked and are changed by participation adjusting human endocrine Kind metabolism, so that delaying human body caducity.
Although the medicinal of aloe, health care are of equal value to be worth to being widely recognized as society.Market has occurred many on makeup The aloe product of product and edible medicine etc., but find no the product of aloe polysaccharide emulsifiable paste class.
The fruit of Cherokee rose (Rosa laevigata Michx.) is traditional Chinese medicine, and the chemical composition of the fruit of Cherokee rose mainly contains apple The effect of fruit and effect acid, citric acid (the effect of lemon and effect acid), tannic acid and resin, still containing saponin, vitamin C.Separately contain Abundant carbohydrate, wherein having reduced sugar 60% (fructose 33%), sucrose 1.9%, and a small amount of starch.Its pharmacological action has three sides Face：1st, the bacteriostasis fruit of Cherokee rose contains tannin, makees bacteriostatic experiment with plate method, 25% decoction is to staphylococcus aureus, large intestine Bacillus, Pseudomonas aeruginosa, clostridium tetani and Leptospira have inhibitory action.Chicken embryo experiment proof, fruit of Cherokee rose decoction convection current Influenza Virus inhibitory action is very strong, and f1 also has A type 57-4 plants of Asia, Lee plants B-mode, 1233 plants of the third type and fourth type celestial platform strain Effect.2nd, study of anti-atherogenic effect rabbit feeding cholesterine and appropriate methylthiouracil is added to produce experimental artery congee Sample hardens, and is treated two weeks and two weeks with the fruit of Cherokee rose, serum ornitrol and beta lipoprotein content significantly reduce, liver and heart fat Calm and atherosclerosis of aorta degree substantially mitigates.3rd, other effect fruits of Cherokee rose, which take orally, can promote gastric secretion, and can make intestines Withered film secretion is reduced, and has astringing to arrest diarrhea effect.
Chinese patent CN201480065835.3 is related to a kind of method being incorporated to of the stabilization process based on by freezing, root Can obtain the pure aloe gel of all natural characteristics containing plant according to this method, gel can with emulsifiable paste, lotion, ointment or The form of ointment is frozen ground, is partly administered, for the normal usage of aloe, but primarily to alleviate various burns, Main function component is aloe gel, and biologically active mainly aloe polysaccharide in aloe gel.
Liposome is made of the continuous lipid bilayer similar to cell membrane of one or more layers concentric arrays, interior Aqueous phase space.Multilamellar liposome (MLV) can be divided into according to the size of liposome and form, small unilamellar vesicle (SUV) and big Unilamellar liposome (LUV) three classes.There is class membrane structure just because of liposome, when reprinting medicine, drug molecule is different Degree it is encapsulated in double-deck intermembranous water phase or in embedded double membrane structure, medicine is not easy by enzyme destruction or body fluid Antibody neutralizes, so as to improve the targeting and validity of medicine in vivo.Liposome is used to inject to be popularized very much already, and nearly ten Nian Lai, the research of liposome transdermal delivery system have caused the very big concern of pharmacy circle.Liposome is in topical application major embodiment The effect of five aspects：1st, hydrophilic or lipophilic medicament can be preferably wrapped up, as the carrier of insoluble drug, is had to medicine Compatibilization；2nd, liposome can encapsulate the biopharmaceutical macromolecular drug for being difficult to transdermal medicine and being easy to be destroyed by intestines and stomach, promote Into its Transdermal absorption, whole body therapeutic effect is produced；3rd, liposome, the liposome and skin that lipoid particularly similar to sebum is formed Skin Stratum corneum lipids have the similitude of height, can increase accumulation of the medicine in local skin, so as to play lasting insoluble drug release Effect；4th, medicine is strengthened into the effect in the lipoid of cuticula or epidermis；5th, there is the speed limit envelope barrier of medicine systemic Absorption Effect, reduces the systemic Absorption of medicine, so as to avoid the toxic side effect of medicine.
Mechanism on liposome local skin medication application may have the following aspects：1st, lipid physical efficiency entering angle matter The deep of layer, or even the skin corium under epidermis, are conducive to the medicine in local storage higher concentration, enhancing medicine is in part Activity；2nd, liposome and phospholipid molecule cannot pass through complete skin；3rd, liposome made of skin lipoid, than made of phosphatide Liposome is easier to enter keratoderma, the reason is that the composition of " sebum " liposome is similar to cuticula class lipid bilayer； 4th, liposome bilayers structure, is the necessary condition for reaching effective skin transhipment, some researches show that environmentally friendly mycin phosphatide fat The percutaneous absorbtion amount of plastid is more far beyond its emulsion；5th, transmission characteristic of the liposome in skin is different because of composition, containing insatiable hunger With the liposome (liquid crystal state) made by the phosphatide of aliphatic acid, the liposome made by the phosphatide of more saturated aliphatic acid, has more Intradermal penetration；6th, liposome is different because of preparation process in intracutaneous permeability, it may be possible to due to technique influence liposome into Layer property, uniformity, size, these factors or its combined influence determine skin and the interactive active surface of liposome, from And influence liposome and enter skin difficulty or ease；7th, the source of skin is different, and the uptake of lipoid is different, and medicine transit dose is different；8、 The liposome of water soluble drug can also enter deep skin quickly, and its aqueous solution cannot enter；9th, pilosebaceous follicle may It is the main thoroughfare that liposome enters deep skin；10th, liposome can make cuticula humidifying, and aquation is strengthened, and makes cutin thin The structural change of intercellular, hydrophobic tail is disorganized in double-layer of lipoid, and liposoluble medicine can be acted on by diffusion and capillary attraction, Into space between cells；11st, the liposome fusion of the phosphatide of liposome and cuticula, changes Stratum corneum lipids the Nomenclature Composition and Structure of Complexes, shape Into a kind of flat nutty structure, by the gap of lipid granule, liposomal encapsulated medicine can enter skin.
Aloe polysaccharide is a kind of glycoprotein, can be according to its hydrophobic power, or incorporation double-layer of lipoid, or is wrapped in water mutually sky Between, or with diversified forms such as fat component covalent bonds it is assembled into liposome.
The fruit of Cherokee rose is extracted from natural plant, and fruit is won after the Frost's Descent, it is formed for holder development, interior tool fruit.Holder Expand as ampuliform, 2.5-4 centimetres long, 0.8-2 centimetres of diameter, outer surface rufous, micro- tool gloss are de- by most spinigers Fall behind existing coarse dentation；The micro- curling of base portion, handle in having sometimes, middle part is plump, and the micro- excessive contracting in top, top is open and flat, and there is Huang in center Color base for post.Visible inner wall has many light yellow fine hair after holder is cut.30-50 containing achene, present irregular avette or ellipse It is circular and flat, there are 3-4 angles rib and a zanjon.Pericarp is faint yellow, wooden hard, outer by the faint yellow fine hair to come off, with fruit point One end it is special more.
Flavones content is very high in cherokee rose fruit, shows that this wild plant resource has very big Development volue.Jin Ying Seed total flavonoid class compound is mainly two class of flavones and flavonols, wherein what is separated and determine structure has Quercetin, rutin, kaempferia galamga Phenol, cyanidenon, apiolin, dihydro apiolin, glycyrrhizin, potengriffioside A and 4 ＇, 5,7- trihydroxyflavone alcohol -3-O- β-D- Ten several flavone compounds such as (6 ＇ ＇-O (E)-p- hydroxyls benzene acryloyl) glucopyranoside.Many flavone compound tools Antiviral activity and anti-inflammatory activity, it is now reported to there is Kaempferol and Chrysin to suppress herpes simplex virus, human coronary's disease Poison, human rotavirus replicate.Quercetin, quercitin, isoquercitrin can substantially suppress herpes simplex virus HSV-1 and HSV- 2 virus infection, inhibiting rate is up to 50% or so.The activation that apiolin, Chrysin can suppress HIV-1 passes through new mechanism.Rutin pair A/PuertoRico/8/1934(H1N1)、A/FM1/1/47(H1N1)、A/Human/Hubei/3/2005(H3N2)、A/ 4 plants of influenza viruses of Beijing/32/92 (H3N2) are respectively provided with In-vitro Inhibitory Effect, wherein to A/PuertoRico/8/1934 (H1N1) medium effective concentration (EC 50) of strain is minimum, selects index (SI) maximum, pharmacy in vitro is best.Cyanidenon convection current The propagation of Influenza Virus has stronger inhibitory action, while also can effectively suppress the expression of inflammatory cytokine, influences TLR signals and leads to Road and the expression of Apoptosis-Related Factors, so as to mitigate immunologic mjury of the virus to host cell, reduce Apoptosis, play anti- The effect of virus.
Two phase aqueous extraction system (aqueous two-phase systems, ATPS) refers to two kinds of hydrophilic compounds Two not miscible aqueous phase systems that aqueous solution spontaneously forms after being mixed under certain mass fraction.With conventional solvent extraction Principle is similar, i.e., target substance is on two alternate selectivity distribution and intermolecular hydrogen bonding effect, Van der Waals force, hydrophobic effect, boundary The factors such as surface properties are related.Aqueous two-phase extraction is a kind of new liquid-liquid extraction and separation technology, it is easy to operate, separative efficiency is high, point Easily amplify from mild condition and technique, possess the prospect of industrial separation natural products.At present, existing extraction system has height Polymers-high polymer-water, high polymer-salt-water, ionic liquid-salt-water, Small molecule organic solvents-salt-water etc., are chiefly used in albumen The extraction of the bioactive substances such as matter, enzyme, amino acid, antibiotic, polysaccharide.
Traditional extraction system is often cooperatively formed with polyethylene glycol and salt, and the cost is relatively high for high polymer, is difficult to recycling again Utilize.If cooperatively forming extraction system with low toxicity, ethanol and ammonium sulfate cheap, easily recycle, extraction cost will be undoubtedly reduced And toxicity, while overcome traditional double-aqueous phase system phase separation time length, easy the shortcomings of emulsifying.In recent years, the double water of ethanol-ammonium sulfate Phase system is widely used to the extraction of active ingredient in natural plants.Chlorination is added in ethanol-ammonium sulfate double-aqueous phase system Sodium, can shorten the split-phase time, improve phase separation speed.Ethanol-ammonium sulfate double-aqueous phase system of addition sodium chloride is had no at present Report applied to the extraction of fruit of Cherokee rose general flavone.
It at home and abroad there is no the dynamic of aloe polysaccharide and the anti-monovesicle viral bleb preparation of fruit of Cherokee rose general flavone compound external application Thing pharmacological evaluation is reported.
The content of the invention
It is an object of the present invention to provide a kind of lipidosome cream of anti-herpesvirus, it contains aloe extract and gold Cherry seed total flavonoid extract, is easily absorbed by the skin, and has the preferable effect of drugs of anti-herpesvirus.
It is a further object to provide the preparation method of the lipidosome cream of above-mentioned anti-herpesvirus, this hair is realized Bright product industrialization.
According to an aspect of the present invention, using aloe extract and fruit of Cherokee rose extractive of general flavone as core ingredient, then help With a series of auxiliary material, the good lipidosome cream of permeability is obtained, it can be used as preferable anti-herpesvirus liniment product；
According to another aspect of the present invention, the present invention is gone out with protecting core ingredient active with the theory of promotion Cutaneous permeation Hair, coordinates addition and the sequence of operations program of auxiliary material, successfully prepares good penetrability, the lipidosome cream production being easily absorbed by the skin Product.And by being contrasted with similar product, the validity and the verification of otherness experimental study of this product and technique are carried out.
The present invention uses the complex liposome containing aloe extract and fruit of Cherokee rose extractive of general flavone, with emulsifiable paste matrix system It is standby into cream form, it is possible to increase liposomal encapsulated property, improve aloe extract and fruit of Cherokee rose extractive of general flavone activity and Drug effect, while there is the advantages of preparation is simple, easy to use.
Aloe extract (main component is aloe polysaccharide, and aloe polysaccharide content is 0.1-1mg/ml) is being prepared, and is being prepared It is necessary to by aloe extract and the fruit of Cherokee rose after fruit of Cherokee rose extractive of general flavone (general flavone content is 0.8mg/ml~1.4mg/ml) Extractive of general flavone is compound, is coated in membrane material and forms liposome, this operating procedure is very crucial.
Since aloe polysaccharide is a kind of macromolecular substances with biological activity, it is easily affected by other factors and Activity is reduced, therefore during complex liposome is prepared, we are while considering that protein conformation is not subject to destroy, also In view of being protected aloe polysaccharide and the bioactivity of fruit of Cherokee rose extractive of general flavone as far as possible.
Extraction for plant medicinal materials, traditional extraction system are often cooperatively formed with polyethylene glycol and salt, high polymer The cost is relatively high, is difficult to recycling.If extractor body is cooperatively formed with low toxicity, ethanol and ammonium sulfate cheap, easily recycle System, will undoubtedly reduce extraction cost and toxicity, while overcome traditional double-aqueous phase system phase separation time length, easily emulsification etc. to lack Point.Sodium chloride is added in ethanol-ammonium sulfate double-aqueous phase system, the split-phase time can be shortened, improves phase separation speed.
The present invention prepares liposome by film dispersion method.Different membrane material formulas is compared, obtains ideal form Liposome.Present invention determine that scheme be that membrane material is used as using the soybean lecithin of domestic natural material and cholesterol, is added at the same time Enter micro stearmide, carry positive charge to adjust membrane material, by the use of appropriate vitamin E as antioxidant, made to avoid membrane material It is standby and storage during oxidation deterioration, by the pH value of aloe extract and the water phase of fruit of Cherokee rose extractive of general flavone be adjusted to for 7.2, to ensure that polysaccharide molecule carries the negative electrical charge opposite with membrane material, so as to improve the envelop rate of aloe polysaccharide.
Pharmaceutical carrier of the emulsifiable paste matrix as liposome, for stablizing major ingredient, uses Tween-80 conduct in emulsifiable paste matrix Emulsifying agent, coordinates the vitamin E in liposome to carry out stabilization to aloe polysaccharide, can reduce aloe polysaccharide in lipidosome cream Leakage, improve the stability of liposome, improve drug effect.
Make since the protective effect of keratoderma barrier causes usual cream to hardly enter human skin depths and play With, cause outside the skin that most of medicine refused, thus to reach the blood concentration of whole body therapeutic effect be highly difficult.It is conventional Way be to improve the concentration of medicine, dose increases, this virtually causes the waste of medicine, so that production cost occupies It is high not under.
Therefore a crucial problem for needing to solve in above-mentioned emulsifiable paste matrix preparation process is to improve the permeability of emulsifiable paste.
The present invention from lifting medicine osmotic efficiency start with, on the basis of usual cream matrix add high-efficient penetrant, Natural complex race is so as to not only expand medicinal application scope, it is often more important that can greatly improve the use effect of new product Fruit.
After obtaining lipidosome cream (LFJ emulsifiable pastes), according to preclinical animal studies guidelines, complex liposome has been carried out The zoopery of emulsifiable paste, including the effect experiment for monovesicle viral bleb, toxicologic study and skin irritation test.
Beneficial effect：
The present invention is carried out with the composite liposome body emulsifiable paste containing aloe extract and fruit of Cherokee rose extractive of general flavone first Viral effect experiment, it is as a result preferable, expand the application range of aloe and fruit of Cherokee rose product.
The present invention utilizes liposome by aloe extract (main component is aloe polysaccharide) and fruit of Cherokee rose extractive of general flavone The compound of (main component is flavones) is wrapped in liposome, using can effectively extend aloe polysaccharide and Jin Ying during medicine The action time of son, improves effective bioavilability in vivo.
In the ideal range, emulsifiable paste prepared by the main ingredient after parcel meets quality through inspection will for the liposomal particle size of preparation Ask；Results of animal shows, which can effectively suppress varicella zoster infection, and security meets the requirements, and with general city Sell emulsifiable paste to compare, positive effect has significant difference, better than general aloe emulsifiable paste.
The advantages of Product Process of the present invention is stablized, and product homogeneity is good, and drug effect improves, and main ingredient permeability is high.For epidermis Illness, it is easy to use, be applied directly to lesion.
Brief description of the drawings
Fig. 1：Standard curve
a:Aloe polysaccharide standard curve b：Fruit of Cherokee rose general flavone standard curve；
Fig. 2：The grain size distribution of sample 2；
Fig. 3：The grain size distribution of sample 8；
Fig. 4：The electron microscopic morphology figure of sample 1；
Fig. 5：The electron microscopic morphology figure of sample 2；
Fig. 6：The electron microscopic morphology figure of sample 3；
Fig. 7：The electron microscopic morphology figure of sample 4；
Embodiment
The present invention is described in further detail with reference to embodiment, but the scope of the invention is not limited to these realities Apply the scope of example.
Material source：
Aloe (Aloe vera var chinesis (haw.) Berg.)：Strain the Chinese aloe aloe, Shenzhen Polytechnic Plant division is planted after Li Xiaodong professor identifies confirmation, place of production Shenzhen City, Guangdong Province.Experiment is using no disease and pests harm, free of contamination health Fresh leaf of aloe.
The fruit of Cherokee rose (Rosa laevigata Michx.)：The fruit of Cherokee rose removes calyx for cherokee rose fruit, splits and removes kind The fruit of Cherokee rose meat of son, purchased from Shenzhen Neptune occasion pharmacy, place of production Jiangxi Fuzhou City.
Term explanation：In the present invention, the main component of aloe extract is aloe polysaccharide, and Fructus Rosae Laevigatae extract is golden cherry Seed total flavonoid；
Soybean lecithin abbreviation SPC, cholesterol abbreviation CHOL, vitamin E abbreviation VE.
Embodiment 1【The preparation of aloe polysaccharide extract】
1.1 aloe polysaccharide is extracted using water extraction and alcohol precipitation method：500 grams of aloe is taken, is cleaned, is removed blade tip and tea residue, distilling Soaked 30 minutes in water, remove the yellow juice of surface exudation.Then epidermis is cut, internal layer gel is placed in beaker, adds 3 Times distilled water, when being put into that hot dipping extraction 4 is small in 55 DEG C of thermostat water baths.Leaching liquor 2500r/min centrifuges 5min, Bu Shi leakages Struggle against filtered off with suction.The filtrate Rotary Evaporators being obtained by filtration are concentrated under reduced pressure, with HCl tune pH3.2 or so, are slowly added into 95% Ethanol, side edged stir, and continue about 15-30 minutes.When quiescent setting 12 is small at room temperature, 2500r/min centrifuges 7min, obtains To polysaccharide precipitation.
Precipitation is washed respectively with ethanol, acetone and ether successively, and washing is three times, molten after dry 8h in vacuum desiccator altogether Solution obtains aloe polysaccharide extracting solution in 250ml distilled water.
The extracting solution aloe total sugar content that 1.2 measure 1.1 obtain：Precision weighs 0.100g glucose and (is dried extremely at 105 DEG C Constant weight) it is placed in 100ml volumetric flasks, add distilled water constant volume.The accurate 5ml solution that measures is placed in from 100ml volumetric flasks again In 50ml volumetric flasks, add distilled water constant volume.Among 50ml volumetric flasks respectively it is accurate draw 0ml, 1ml, 2ml, 3ml, 4ml, 5ml is placed in 10ml volumetric flasks, adds distilled water constant volume respectively.It is 0mg/ml, 0.01mg/ml, 0.02mg/ to respectively obtain concentration The glucose standards solution of ml, 0.03mg/ml, 0.04mg/ml, 0.05mg/ml.
By above-mentioned glucose standards solution, for the accurate 2ml that draws in different test tubes, each test tube adds phenol respectively in order 1ml shakes up, enriching sulfuric acid 5ml, after shaking up rapidly, stands 5min, and boiling water heating water bath 15min, is cooled to room temperature, and is tried with zero pipe Sample adjusts zero point, and absorbance is surveyed at 490nm wavelength, and measurement result is as shown in table 1.Draw standard curve and obtain regression equation y= 16.083x+0.0088 linearly dependent coefficient R2=0.999, the standard curve for measuring aloe polysaccharide is shown in attached drawing 1a.
Table 1：Glucose standard curve initial data
Concentration of glucose (mg/ml) 0 0.01 0.02 0.03 0.04 0.05
Absorbance A 0 0.179 0.324 0.501 0.658 0.803
Aloe polysaccharide extracting solution is taken, is filtered, precision measures filtrate 10mL, is settled to 100mL, is diluted up to aloe polysaccharide Liquid.Precision measures aloe polysaccharide dilution liquor 2mL and is placed in 10mL volumetric flasks, adds distilled water to dissolve and is diluted to scale.From Middle precision pipettes 2mL and is placed in test tube, according to the processing method of glucose standards solution, is operated successively from being added phenol, with zero pipe Adjust zero point, absorbance is 0.26 at 490nm wavelength, according to regression equation calculation multiplied by extension rate 50, calculate aloe is more Aloe polysaccharide content is 0.781mg/ml in sugared extract concentration.
Embodiment 2【The preparation of fruit of Cherokee rose extractive of general flavone】
2.1 alcohol refluxs-aqueous two-phase extraction prepares fruit of Cherokee rose general flavone extracting solution：It will be carried out after 103 DEG C of drying of the fruit of Cherokee rose It is spare that mechanical lapping obtains fruit of Cherokee rose powder.By (60~90 DEG C) of the petroleum ether reflux degreasing 2 times of fruit of Cherokee rose powder, solid-liquid ratio 1: 15 (g：ML), each degreasing time 3h, filters to obtain filter residue, and it is spare that fruit of Cherokee rose degreasing product is obtained after 60~80 DEG C of dryings.Weigh Jin Ying Sub- degreasing product 25g, adds 60% ethanol solution 500mL, 60 DEG C of refluxing extractions 3 times, each 5h, merging filtrate, after being concentrated under reduced pressure Being settled to 500mL with distilled water, to obtain fruit of Cherokee rose general flavone crude extract spare.
Aqueous two-phase cumulative volume is set as 500mL.Fruit of Cherokee rose general flavone crude extract 110mL is drawn (equivalent to the body of aqueous two-phase Fraction 22%), be placed in glass container, add absolute ethyl alcohol 135mL (being 27% equivalent to aqueous two-phase volume fraction) and Ammonium sulfate (so that double-aqueous phase system ammonium sulfate containing 0.21g/mL), the 20g sodium chloride of 105g, with 2% hydrochloric acid solution and 2% hydrogen-oxygen Change sodium solution regulation system pH value 4~5, abundant shaken well stands 10min, and two-phase reaches phase separation.Upper liquid is separated, is subtracted 250mL is settled to 30% ethanol as solvent after pressure concentration and obtains fruit of Cherokee rose general flavone extracting solution.
The extracting solution general flavone content that 2.2 measure 2.1 obtain：It is (dry at 120 DEG C that precision weighs rutin standard items 10.5mg To constant weight), 50mL is settled to after being dissolved with 60% ethanol, the rutin standard items that 0.20mg/mL is made use solution.Precision is drawn 0th, the rutin standard items of the 0.21mg/mL of 1.0,2.0,3.0,4.0,6.0 and 8.0mL use solution, are respectively placed in 25mL capacity In bottle.5%Na NO are added in volumetric flask2Solution 1mL, mixes, and 10%Al (NO are added after standing 6min3)3Solution 1mL, is mixed It is even, 4%Na OH solution 10mL are added after standing 6min, add 60% ethanol solution to be settled to scale, are mixed, stand 15min.With Zero pipe adjusts zero point, and absorbance is surveyed at wavelength 510nm, draws standard curve, obtains regression equation y=10.227x-0.0065, Linearly dependent coefficient R2=0.9994, the standard curve for measuring general flavone is shown in attached drawing 1b.
Table 2：Rutin standard curve initial data
Rutin concentration/(mg/mL) 0 0.0084 0.0168 0.0252 0.0336 0.0504 0.0672
Light absorption value A 0 0.068 0.164 0.257 0.34 0.506 0.681
Fruit of Cherokee rose general flavone extracting solution is taken to filter, precision measures filtrate 10mL, is settled to 100ml, always yellow up to the fruit of Cherokee rose Ketone dilution.Precision measures fruit of Cherokee rose general flavone dilution 6mL, is placed in 25ml volumetric flasks, according to 2.2 standard curve determination sides Method develops the color, and adjusts zero point with zero pipe, it is 0.319 that absorbance is measured at wavelength 510nm, and it is total to obtain the fruit of Cherokee rose according to regression equation calculation General flavone content is 1.33mg/ml in flavone extractive.
2.3 aqueous two-phase extractions prepare the screening of fruit of Cherokee rose general flavone extracting solution extraction conditions
General flavone extraction yield (y) calculates according to the following formula
Y=m1/ (m1+m2) × 100% (1)
In formula:The quality of general flavone in m1-upper phase alcohol layer, mg,
The quality of general flavone, mg in m2-lower phase water layer.
(1) when ammonium sulfate mass concentration is 0.22g/mL, fruit of Cherokee rose extracting liq fraction is 20%, volume fraction of ethanol Influence to extraction yield is shown in Table 3.
Table 3：Influence of the volume fraction of ethanol to extraction yield
Volume fraction of ethanol/% 22 23 24 25 26 27 28 29
Extraction yield/% 63.3 64.6 67.2 73.1 78.8 82.4 79.7 77.5
(2) when volume fraction of ethanol 27%, fruit of Cherokee rose extracting liq fraction is 20%, and ammonium sulfate mass concentration is to extraction The influence of rate is shown in Table 4.
Table 4：Influence of the ammonium sulfate mass concentration to extraction yield
(3) when volume fraction of ethanol 27%, ammonium sulfate mass concentration is 0.22g/mL, fruit of Cherokee rose extracting liq fraction pair The influence of extraction yield is shown in Table 5.
Table 5：Influence of the fruit of Cherokee rose extracting liq fraction to extraction yield
Fruit of Cherokee rose extracting liq fraction/% 14 16 18 20 22 24 26 28
Extraction yield/% 72.5 73.1 80.2 85.3 85.1 84.8 79.8 73.5
Embodiment 3【The preparation of complex liposome】
3.1 liposome preparation programs
Preparation process：Membrane material is dissolved in diethyl ether solution becomes organic phase, is added to and contains what is necessarily matched in organic phase Fruit of Cherokee rose general flavone, then under conditions of 40-43 DEG C, the situation of inflated with nitrogen, the rotary speed of Rotary Evaporators is set as 120 revs/min, last organic solvent volatilizees to obtain liposome membrane, then (will be more containing the aloe necessarily matched by aqueous phase solution Sugar adds aqueous dissolution to obtain the final product) it is added in Rotary Evaporators eggplant type bottle, in the case of inflated with nitrogen, in 34-37 DEG C of feelings Under condition, rotary speed is 120 revs/min, carries out vortex oscillation, in the case of water bath sonicator instrument, liposome membrane is crushed, Obtain four kinds of liposome aqueous solutions, carry out filtration sterilization with the filter membrane of 0.45um and 0.2um respectively, then with gel chromatography column into Row concentration, then is carried out with 0.2um films degerming, carry out assay approval and dispense preservation afterwards.
Concrete operations are as follows：
(1) by the use of ether as solvent, compound concentration is 30mg/ml soybean lecithins (SPC) solution 10ml, 4mg/ml cholesterol (CHOL) solution 10ml, 1% vitamin E (VE) solution 5ml, add the fruit of Cherokee rose general flavone powder that 15ml is independently extracted.
(2) by above-mentioned solution be placed in the eggplant-shape bottle of rotary evaporation instrument be evaporated under reduced pressure to bottle wall formed one layer of liposome it is thin Film.
(3) 20ml ether is added, after solubilizing lipids film, adds the homemade aloe polysaccharide aqueous phase solutions of 25ml, mixing gained Solution forms two-phase system.
(4) every group of ultrasound forms single_phase system (about 2-5min) to mixture on water-bath type processor for ultrasonic wave, can put Put 30min not split-phases.
(5) by mixed liquor on a rotary evaporator evaporation under reduced pressure removed organic solvent to gel-forming.
(6) continue 5~10min of reduction vaporization, form aqueous suspension, that is, Liposomal suspensions.
(7) after suspension is formed, continue to dry 5~10min on an evaporator, further remove residual organic solvent.
(8) last inflated with nitrogen to ether taste disappears.Liposome solutions press 2m1/ bottles after will be degerming, are divided in cillin bottle； It is freeze-dried.
The detection of 3.2 liposomes
3.2.1 the granularmetric analysis of liposome
Four groups of complex liposomes prepared by embodiment 3.1 are carried out with the investigation of the indexs such as Morphologic observation, granularmetric analysis, shape State observation is mainly from the form of liposome particles (from being irregularly divided into 1~5 point to mellow and full, more mellow and fuller score value highest).
At room temperature, suitable sample 1000ml distilled water is diluted, with laser diffraction particle size analyzer to obtained The particle diameter of liposome is analyzed, measures its particle diameter distribution and average grain diameter.The automatically derived lipid of Particle Size Analyzer software kit The correlation values such as the grain size distribution of body, average grain diameter, span.
Quality standard in terms of liposomal particle size is as follows：Particle diameter distribution should be normal distribution, and average grain diameter should be 200~ 210nm, span should be 0.80~1.10, and uniformity should be not higher than 0.35.
3.2.2 the morphological observation of liposome
With transmission scanning electron microscope, the form of liposome is observed using negative staining, operating method is as follows：Take lipid The solution of body is appropriate, dilutes 10 times with the PBS buffer for preparing its aqueous phase solution same concentrations and ionic strength, drips to and be placed in filter On copper mesh on paper, 1% phosphotungstic acid of a drop same pH is added according to the pH drops man of PBS buffer, low-temperature heat drying, takes The copper mesh, is observed and taken pictures with transmission scanning electron microscope.
3.2.3 the measure of envelop rate
Complex liposome An Ma West Asias company Sepharose G100 calibrating chromatographic columns are filtered, collect liposome peak, First peak of purifies and separates is liposome peak, and second peak is free drug peak, as follows
Computational envelope rate EE=(B × H)/A × 100%
Wherein:A=upper prop prodrug concentration of liposomes (mg/ml)；The liposome of the excessively complete calibrating columns of B=collects the medicine at peak Concentration (mg/ml)；Medicinal liposome collects the volume at peak and the complex liposome solution body before upper calibrating column after H=crosses calibrating column Long-pending ratio.
3.3 liposome recipe determination schemes
3.3.1 the screening of different membrane material proportionings
We according to the present embodiment 3.1 processing step, except the membrane material ratio among organic phase is different, the reed in water phase Luxuriant growth polysaccharide and fruit of Cherokee rose concentration fix tentatively and are unified for 1.0mg/ml：2.0mg/ml, in the case that other technological parameters are identical, prepares fat Plastid, passes through electron microscopic morphology detection and particle diameter distribution two indices score respectively (score value is the higher the better), it is determined that optimal Membrane material formula, the results are shown in Table 6.
Table 6：
Particle diameter distribution is more concentrated, and score is higher, is scored at 1-5 points；Electron microscopic morphology is more mellow and fuller, and score is higher, is scored at 1-5 Point.The electron microscopic morphology figure of sample 1- samples 4 is shown in that the grain size distribution of Fig. 4-Fig. 7 samples 2 is shown in Fig. 2 respectively, and the score of sample 2 is most It is high.
3.3.2 the screening of different aqueous pH values
We according to the present embodiment 3.1 processing step, in the optimal membrane material ratio situation that embodiment 3.3.1 is selected Under, in the case that other technological parameters are identical, using the aqueous phase system of the buffer solution configuration medicine of different pH value, and prepare load medicine Liposome, separates liposome by gel chromatography, measures the index of envelop rate the optimal of aqueous phase solution is determined PH value, the results are shown in Table 4.
Table 7：
Sample number into spectrum Sample 6 Sample 7 Sample 8 Sample 9 Sample 10
Aqueous pH values 6.4 6.8 7.2 7.6 8.0
Envelop rate 35.2% 39.3% 43.5% 40.2% 36.7%
From the results of view, the pH value of aqueous phase solution belongs to ideal pH value for 7.2.The grain size distribution of sample 8 is shown in Fig. 3.
3.3.3 the selection of different pharmaceutical concentration
We, in the optimal membrane material ratio that embodiment 3.3.1 is selected, implement according to the processing step of the present embodiment 3.1 In the case of the optimal pH that example 3.3.2 is selected, in the case that other technological parameters are identical, medicine is configured using different pharmaceutical concentration The aqueous phase system of product, by measuring the envelop rate of liposome, to investigate bearing capacity situation of the liposome to drug concentration, the result is shown in Table 5.
Table 8：
Sample number into spectrum Sample 11 Sample 12 Sample 13 Sample 14 Sample 15
Aloe polysaccharide (mg/ml) 0.05 0.1 0.5 1.0 2.0
Fruit of Cherokee rose general flavone (mg/ml) 0.25 0.5 1.0 2.0 3.0
Envelop rate 20.1% 36.9% 43.5% 40.7% 23.4%
Pass through above-mentioned experimental result, it is contemplated that the complexity of the extraction process of aloe polysaccharide and fruit of Cherokee rose general flavone, I Among the aqueous phase system thought, the concentration of preferable aloe polysaccharide and fruit of Cherokee rose general flavone respectively 0.1-1.0mg/ml with 0.5-2.0mg/ml。
Embodiment 4【The preparation and detection of emulsifiable paste】
The preparation of 4.1 emulsifiable paste matrixes
The matrix formulations of emulsifiable paste are as shown in table 9.
Table 9：
The step of experimental group for being below 1 with numbering is demonstrated, and illustrates operating process, the experimental group of other numberings is identical, The difference is that the dosage of various composition is variant as shown in table 9.
A, oil phase and water phase are prepared respectively.
Water distribution phase：Take 100ml beakers add glycerine 16ml, dextran 1 g, 3,7- dimethyl octyl group propionic ester 0.15g, Ethyl lactate 1ml, purified water 54ml.
With oil phase：Take 100ml beakers add single stearic glyceride 10g, soybean lecithin 1g, albolene 10g, tween- 80 3ml。
B, by water phase and oil phase, water-bath is dissolved at least more than 80 degree of water temperature.
C, water phase and oil phase are poured over while hot and are mixed together, the stirring of side bevelling, pays attention to having to stir in same direction Mix, as, according to stirring clockwise, used stirring clockwise always with glass bar.
D, after whole water phases and whole oil phases pour into, continue even speed stirring, stop when being cooled to 45 DEG C or so Stirring.
E, as temperature reduces gradually, emulsifiable paste becomes milky gradually.
Operated more than and obtain emulsifiable paste matrix, the results are shown in Table 10 for emulsification test.
Table 10：
It can be seen that the 3rd group, i.e. single stearic glyceride of addition 14% is most suitable for.
The preparation of 4.2 emulsifiable paste finished products (LJF emulsifiable pastes)
The material of use is as follows.
Oil phase：Single tristearin glyceride 14g, soybean lecithin 1g, albolene 10g, Tween-80 3ml.
Water phase：Glycerine 16ml, dextran 1 g, 3,7- dimethyl octyl group propionic ester 0.15g, ethyl lactate 1ml, complex liped Plastid 2g, purified water 50ml.
Concrete operation step：
Ith, by water phase and oil phase, water-bath is dissolved at least more than 80 degree of water temperature.
IIth, treat that oil phase substance and aqueous phase substance are completely dissolved, 1g liposomes are added into aqueous phase substance and are stirred evenly, while hot (being not less than 70 degree), then water is mutually poured slowly into in oil phase
IIIth, side bevelling stirs, and pays attention to having to stir in same direction, as with glass bar according to stirring clockwise Words, always with stirring clockwise.
IVth, after whole water phases and whole oil phases pour into, even speed stirring, when being cooled to 45 DEG C or so, perfuming are continued Essence, stirs evenly, and as temperature reduces gradually, emulsifiable paste becomes milky gradually.
The stable quality detection of 4.3LJF emulsifiable pastes
Stability of the LJF emulsifiable pastes at 4 DEG C, 24 DEG C and 36 DEG C is detected respectively, is preserved to 24 months, using Chinese Pharmacopoeia as mark Whether standard, detection emulsifiable paste appearance meet standard；And detect the release rate Detection of Stability of medicine.As a result it is as shown in the table.
Table 11：LJF emulsifiable pastes physical and chemical stability detects
The results show LJF emulsifiable pastes all meet appearance standard.
Table 12：The release rate Detection of Stability of medicine in LJF emulsifiable pastes
1 month 3 months 6 months 9 months 12 months 15 months 18 months 21 months 24 months
4℃ 92% 90% 90% 90% 90% 90% 90% 89% 90%
24℃ 95% 96% 95% 95% 96% 94% 95% 96% 94%
36℃ 94% 93% 93% 94% 93% 93% 93% 93% 92%
The results show LJF emulsifiable paste release rates have good stability.
Embodiment 5【The animal experiment of LJF emulsifiable pastes】
5.1 pharmacodynamic study
Using cavy as subjects, the dosage with 5mg/ only, using emulsifiable paste finished product, per 2 smearing affected parts of bu, connects respectively It is continuous to use 7.In positive controls, with the dosages of 6.0mg/ only, in the same fashion using acyclovir.
The anti-cavy inoculation property herpe simplex result of the test (X ± SD) of table 13.LJF emulsifiable pastes
Compared with model group：*P<0.05, * * P<0.01, * * * P ﹥ 0.05, should be the result shows that the LJF breasts of three groups of different ratios Cream (aloe polysaccharide 0.1mg：Fruit of Cherokee rose general flavone 0.5mg, aloe polysaccharide 0.5mg：Fruit of Cherokee rose general flavone 1mg；Aloe polysaccharide 1mg： Golden ` cherries seed total flavonoid 2mg) there is obvious inhibiting effect to guinea pig experimental monovesicle viral bleb, and its intensity is more with aloe The content of sugar and fruit of Cherokee rose general flavone increases and strengthens, LJF emulsifiable pastes (aloe polysaccharide 1mg：Fruit of Cherokee rose general flavone 2mg) group * * P< 0.01, and traditional chemical drugs acyclovir comparative advantages are obvious, even if the LJF emulsifiable pastes (aloe 0.1 using minimum metering：Gold Cherry 0.5mg/ is only) dosage, therapeutic effect is also superior to control positive drug, * P<0.05.It can be seen at the same time by experiment same Under dosage and proportioning, only because the drug effect for not adding penetrating agent and vitamin E is substantially not so good as normal group, * * * P ﹥ 0.05 added, Illustrate the important drug action of penetrating agent and vitamin E, the emulsifiable paste result of the test of independent component illustrates that same dose of aloe is more The independent component effect of sugar or the total brass of the fruit of Cherokee rose is not so good as the effect of same dose of compound cream.
5.2 pharmacological research
Using mouse as experimental subjects, the dosage with 1mg, 3mg, 10mg/ only, takes one group of LJF emulsifiable paste (aloe polysaccharide respectively 1mg：JINYINZI flavones 2mg) its damaged skin is smeared, observe the influence to its autonomic activities；
Using anesthetized cat as experimental subjects, the dosage with 0.5g, 1g, 5g/ only, takes LJF emulsifiable pastes to smear its damaged skin respectively, Observe the influence to its blood pressure, heart rate, respiratory rate, depth of respiration and electrocardiogram；Anesthetized cat damaged skin smears the emulsifiable paste 0.5g, 1g, 5g/, the results show that being had no significant effect to its blood pressure, heart rate, respiratory rate, depth of respiration and electrocardiogram, specifically It see the table below.
Table 14：Influence (X ± SD) of the LJF emulsifiable pastes to mouse autonomic activities
The result shows that LJF emulsifiable pastes have no significant effect the autonomic activities of mouse.
Table 15：Influence (X ± SD, n=6) of the LJF emulsifiable pastes to anesthetized cat heart rate
Table 16：Influence (X ± SD, n=6) of the LJF emulsifiable pastes to anesthetized cat systolic pressure:
Table 17：Influence (X ± SD, n=6) of the LJF emulsifiable pastes to anesthetized cat diastolic pressure
Table 18：The influence (X ± SD, n=6) of LJF emulsifiable paste anesthetized cat respiratory rates
Table 19：The influence (X ± SD, n=6) of LJF emulsifiable paste anesthetized cat depths of respiration
5.3 toxicologic study
5.3.1 rat acute toxicity test
With 20gkg-1LJF emulsifiable paste maximum dosage-feedings, continuous observation 7 days.
1.1 × 10 of the dosage equivalent to clinical dosage, does not also occur abnormal response, in none death of rat in 7 days4Times, So clinical application is very safe.
5.3.2 rat chronic toxicity test
LJF emulsifiable pastes are taken to be administered by 15g/ dosage at rat damaged skin, it is once a day, 28 days continuous (equivalent to facing Bed intends 4 times with the cycle).
As a result compared with the control group, to observed each index, (activity, weight, dietary amount, amount of drinking water, blood routine, blood are given birth to Change index, organ coefficient, histopathological examination) equal Non Apparent Abnormality.Successive administration withdrawal observation 14 days again after 28 days, it is above-mentioned Each equal Non Apparent Abnormality of index.
5.5.3 security inspection
5.5.3.1 skin anaphylactic test：
LJF emulsifiable pastes press 10g/g, and 0.2g/ dosage, contacts the allergic reaction such as skin, no redness, necrosis repeatedly.
5.5.3.2 skin irritation test：
LJF emulsifiable pastes press 10g/g, 1.0g/ dosage, smear skin within continuous 7 days, to cavy intact skin and damaged skin without Irritative response.
Above detailed description of the present invention is not intended to limit the present invention, and those skilled in the art can make on this basis Various changes and deformation, without departing from the spirit of the present invention, should all belong to the scope of the claims in the present invention.

Claims (10)

  1. A kind of 1. lipidosome cream of anti-herpesvirus, containing emulsifiable paste matrix and complex liposome, wherein being used to prepare described The material of complex liposome includes：
    Aloe extract, the aloe extract are the water extract-alcohol precipitation extract of aloe, and aloe polysaccharide content is in extract 0.1-1mg/ml。
  2. 2. lipidosome cream according to claim 1, it is characterised in that be used to prepare the material of the complex liposome also Including fruit of Cherokee rose extractive of general flavone, general flavone content is 0.8mg/ml~1.4mg/ in the fruit of Cherokee rose extractive of general flavone ml。
  3. 3. lipidosome cream according to claim 1, it is characterised in that the lipidosome cream contains weight ratio 1: 20-2:1 aloe polysaccharide and fruit of Cherokee rose general flavone.
  4. 4. lipidosome cream according to claim 2, it is characterised in that the fruit of Cherokee rose extractive of general flavone is returned through ethanol Stream-aqueous two-phase extraction is made, and the aqueous two-phase extraction is ethanol-ammonium sulfate double-aqueous phase system；The double-aqueous phase system pH value 4~ 5, wherein sodium chloride concentration control is controlled in the range of 25%~29% in 0.04g/mL, volume fraction of ethanol, ammonium sulfate quality Concentration is controlled in the range of 0.18g/ml~0.23g/ml.
  5. 5. lipidosome cream according to claim 4, it is characterised in that the fruit of Cherokee rose extractive of general flavone is with golden cherry Sub- crude extract is made through alcohol reflux-aqueous two-phase extraction, and the preparation method of the fruit of Cherokee rose crude extract is as follows：
    The fruit of Cherokee rose is dried, grinds to obtain fruit of Cherokee rose powder；By the petroleum ether reflux degreasing of fruit of Cherokee rose powder, filter residue is filtered to obtain, is done Fruit of Cherokee rose degreasing product is obtained after dry；Fruit of Cherokee rose degreasing product is weighed, adds ethanol solution, refluxing extraction, collects filtrate, be concentrated under reduced pressure Obtain fruit of Cherokee rose crude extract.
  6. 6. lipidosome cream according to claim 1, it is characterised in that the weight of the emulsifiable paste matrix and complex liposome Than for 923：40；
    The complex liposome includes liposome membrane material, and the liposome membrane material includes soybean lecithin, cholesterol and vitamin E；
    The emulsifiable paste matrix includes oil phase auxiliary material and water phase auxiliary material, the oil phase auxiliary material include single palmitostearate, tween- 80th, hydrogenated soy phosphatidyl choline and albolene；The water phase auxiliary material includes glycerine, dextran, 3,7- dimethyl octyl group propionic acid Ester and ethyl lactate.
  7. 7. lipidosome cream according to claim 6, it is characterised in that it is 30 that the liposome membrane material, which includes weight ratio,: 4:5 soybean lecithin, cholesterol and vitamin E；
    It is 14 that the oil phase auxiliary material and water phase auxiliary material, which include weight ratio,:3:1:10:16:1:0.15:1 single palmitostearate, Tween-80, hydrogenated soy phosphatidyl choline, albolene, glycerine, dextran, 3,7- dimethyl octyl group propionic esters and ethyl lactate；
    The fruit of Cherokee rose general flavone is present in liposome oil phase, and aloe polysaccharide is present in liposome water phase.
  8. 8. lipidosome cream according to claim 1, it is characterised in that the aloe extract is prepared by following methods：
    Ith, aloe is taken, is cleaned, is removed blade tip and tea residue, soaked in distilled water, removes the yellow juice of surface exudation；Then cut Remove epidermis, internal layer gel be placed in beaker, add 3 times of distilled water, be put into 55 DEG C of thermostat water baths hot dipping extraction 4 it is small when；
    IIth, leaching liquor centrifuges, centrifugal condition 2500r/min, 5min, filtering；Obtained leaf juice is concentrated under reduced pressure.Use again HCl tune pH3.2 or so, the ethanol for being slow added into 95%, side edged stir 15-30 minutes；
    IIIth, centrifuged after when quiescent setting 12 is small at room temperature, centrifugal condition 2500r/min, 7min, obtain polysaccharide and sink Form sediment；
    IVth, washed successively with ethanol, acetone and ether, in triplicate；Be then placed in vacuum desiccator dry 8 it is small when.
  9. 9. lipidosome cream according to claim 1, it is characterised in that the complex liposome is prepared by following methods：
    1) take solvent compound concentration as 30mg/ml soybean lecithin solution 10ml, 4mg/ml cholesterol solution 10ml of ether, 1% vitamin E solution 5ml；
    2) above-mentioned solution is placed in eggplant-shape bottle, is evaporated under reduced pressure using Rotary Evaporators thin to bottle wall one layer of liposome of formation Film；
    3) 20ml ether is added, after solubilizing lipids film, adds the aloe extract of 25ml and the mixed solution of Fructus Rosae Laevigatae extract, The pH of the mixed solution is 7.2, and the solution for mixing gained forms two-phase system；
    4) the ultrasound 2-5min on water-bath type processor for ultrasonic wave, single_phase system is formed to mixture；
    5) by mixed liquor on a rotary evaporator evaporation under reduced pressure removed organic solvent to gel-forming；
    6) continue 5~10min of reduction vaporization, form aqueous suspension, that is, Liposomal suspensions；
    7) continue to dry 5~10min on an evaporator, further remove residual organic solvent；
    8) inflated with nitrogen to ether taste disappears, and degerming rear liposome solutions are pressed 2ml/ bottles, is divided in cillin bottle, it is dry to carry out freezing Dry complex liposome to obtain the final product.
  10. 10. lipidosome cream according to claim 1, wherein the preparation method of the lipidosome cream includes following steps Suddenly：
    (i) take complex liposome 2g, weigh single palmitostearate 14g, Tween-80 3ml, soybean lecithin 1g, Bai Fan respectively Intellectual circle 10g is as oil phase auxiliary material；Glycerine 16ml, dextran 1 g, 3,7- dimethyl octyl group propionic ester 0.15g, breast are weighed respectively Acetoacetic ester 1ml makees water phase auxiliary material；Take purified water 50ml；
    (ii) by water phase auxiliary material and oil phase auxiliary material, water-bath is dissolved at least more than 80 degree of water temperature；
    (iii) treat that oil phase auxiliary material and water phase auxiliary material are completely dissolved, 1g liposomes are added into aqueous phase substance and are stirred evenly, while hot at it During not less than 70 degree, then water is mutually poured slowly into oil phase, be stirred；
    (iv) after whole water phase auxiliary materials and whole oil phase auxiliary materials pour into, continue even speed stirring, when being cooled to 45 DEG C or so, add Essence, stir evenly, and as temperature reduces gradually, emulsifiable paste becomes milky gradually.
CN201710948443.7A 2017-10-12 2017-10-12 A kind of lipidosome cream of anti-herpesvirus and preparation method thereof Pending CN107898903A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710948443.7A CN107898903A (en) 2017-10-12 2017-10-12 A kind of lipidosome cream of anti-herpesvirus and preparation method thereof

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710948443.7A CN107898903A (en) 2017-10-12 2017-10-12 A kind of lipidosome cream of anti-herpesvirus and preparation method thereof

Publications (1)

Publication Number Publication Date
CN107898903A true CN107898903A (en) 2018-04-13

Family

ID=61841368

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710948443.7A Pending CN107898903A (en) 2017-10-12 2017-10-12 A kind of lipidosome cream of anti-herpesvirus and preparation method thereof

Country Status (1)

Country Link
CN (1) CN107898903A (en)

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1660411A (en) * 2004-01-16 2005-08-31 深圳市海王英特龙生物技术股份有限公司 Cream of liposome of interferon
US6946490B2 (en) * 1996-02-12 2005-09-20 Meryl Squires Antimicrobial treatment for herpes simplex virus and other infectious diseases

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6946490B2 (en) * 1996-02-12 2005-09-20 Meryl Squires Antimicrobial treatment for herpes simplex virus and other infectious diseases
CN1660411A (en) * 2004-01-16 2005-08-31 深圳市海王英特龙生物技术股份有限公司 Cream of liposome of interferon

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
杨项权等: ""芦荟在皮肤科临床的应用"", 《北京中医》 *
郑民实等: ""中草药抗HBsAg的实验研究"", 《微生物学杂志》 *

Similar Documents

Publication Publication Date Title
Karimi et al. Phytosome and liposome: the beneficial encapsulation systems in drug delivery and food application
Hooresfand et al. Preparation and characterization of rutin-loaded nanophytosomes
CN101537187B (en) Anthocyanin phospholipids compound and preparation method thereof
EP1092765A2 (en) Germination-activated red Ganoderma lucidum spores and method for producing the same
CN101380390A (en) Preparation method of Pu-Er ripe tea standard extract PRC-001 and preparation method and use thereof
CN1813782B (en) Composition of ginkgo leaf extract and dipyridamole, medicine containing same and preparing method thereof
CN101863871A (en) Total glycosides of Rhodiola rosea, medical application and preparation method thereof
CN102302592A (en) Preparation technology and purpose of citrus grandis peel flavones
Wang et al. Enhancement of oral bioavailability and hypoglycemic activity of liquiritin-loaded precursor liposome
CN106619588B (en) It is a kind of containing Co-Q10 from micro-emulsion type alimentation composition, Preparation method and use
ES2843640T3 (en) Procedure for preparing a plant matrix extract with a non-ionic amphiphilic compound as an extraction aid in aqueous medium
RU2322091C1 (en) Composition of biologically active substances and method for preparing its nanodispersion
Mishra et al. Phyto–vesicles: conduit between conventional and novel drug delivery system
Zolgharnein et al. Determination of cichoric acid as a biomarker in Echinacea purpurea cultivated in Iran using high performance liquid chromatography
Tallam et al. A review on phytosomes as innovative delivery systems for phytochemicals
KR101750616B1 (en) Natural liposome comprising red ginseng, process for the preparation thereof and food, pharmaceutical or cosmetic composition comprising the same
CN105125419B (en) A kind of polyethyleneglycol modified vitamin E liposome facial mask and preparation method thereof
CN107898903A (en) A kind of lipidosome cream of anti-herpesvirus and preparation method thereof
CN102626427B (en) Ginkgo dipyridamole composition and preparation method of preparation thereof
CN103877149A (en) Phospholipid complex containing dracocephalum moldavica total flavonoids, self-emulsifying agent prepared from same and preparation method of phospholipid complex containing dracocephalum moldavica total flavonoids
CN110511294A (en) A kind of preparation method and applications of licorice polysaccharide
CN115252502B (en) Preparation method of Hibiscus sabdariffa extract liposome and liposome thereof
TWI618540B (en) Composition for preventing renal toxicity caused by drug toxicity, preparation method thereof and Its use
JP5294847B2 (en) Moisturizer, whitening agent and slimming agent
Janarny et al. Optimization of ethanol-based extraction of phenolic compounds from edible flowers of Cassia auriculata

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for 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: 20180413