CN108324734A - Application of the water-soluble fullerenes derivates in enhancing normal cell activity - Google Patents
Application of the water-soluble fullerenes derivates in enhancing normal cell activity Download PDFInfo
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- CN108324734A CN108324734A CN201810130575.3A CN201810130575A CN108324734A CN 108324734 A CN108324734 A CN 108324734A CN 201810130575 A CN201810130575 A CN 201810130575A CN 108324734 A CN108324734 A CN 108324734A
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- normal cell
- enhancing
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- activity
- cell
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- XMWRBQBLMFGWIX-UHFFFAOYSA-N C60 fullerene Chemical class C12=C3C(C4=C56)=C7C8=C5C5=C9C%10=C6C6=C4C1=C1C4=C6C6=C%10C%10=C9C9=C%11C5=C8C5=C8C7=C3C3=C7C2=C1C1=C2C4=C6C4=C%10C6=C9C9=C%11C5=C5C8=C3C3=C7C1=C1C2=C4C6=C2C9=C5C3=C12 XMWRBQBLMFGWIX-UHFFFAOYSA-N 0.000 title claims abstract description 126
- 229910003472 fullerene Inorganic materials 0.000 title claims abstract description 123
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/44—Elemental carbon, e.g. charcoal, carbon black
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/52—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an inorganic compound, e.g. an inorganic ion that is complexed with the active ingredient
-
- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/19—Cosmetics or similar toiletry preparations characterised by the composition containing inorganic ingredients
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
- A61K8/41—Amines
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/10—General cosmetic use
Abstract
The present embodiments relate to a kind of application of water-soluble fullerenes derivates in enhancing normal cell activity, more particularly to application of the water-soluble fullerenes derivates in preparing the active health products of enhancing normal cell, cosmetics, wherein the water-soluble fullerenes derivates include：At least one of the fullerene derivate of amido modified fullerene derivate, hydroxyl modified；The enhancing normal cell activity includes：At least one of the proliferation for enhancing the activity of succinate dehydrogenase in the mitochondria of normal cell, enhancing reducing substances activity and quickening normal cell in normal cell.The fullerene derivate of amido modified fullerene derivate, hydroxyl modified has excellent water solubility, and molecular particle size is smaller, easily enters cell interior, remains to the activity for significantly increasing normal cell even at a low concentration.
Description
Technical field
The present invention relates to biomedicine fields, further to water-soluble fullerenes derivates in enhancing normal cell activity
In application more particularly to water-soluble fullerenes derivates in preparing the active health products of enhancing normal cell, cosmetics
Using.
Background technology
The elementide with enclosed construction that fullerene is made of different number of carbon atom, unique structure and
Attention of the performance by people.The water solubility and biocompatibility of fullerene can be significantly improved by functional modification, to
Expand application study of the fullerene in biomedical sector.It is total to currently, water-soluble fullerenes derivates are widely used in nuclear-magnetism
Shake the numerous areas such as radiography, oncotherapy, chemicotherapy protection, cosmetic additive agent, anti-rad and antioxidant.We go deep into
Influence of the water-soluble fullerenes derivates to cell activity is had studied, for further pushing it in the application of biomedical sector
Research is of great significance.
Being disclosed in the information of the background technology part, it is only intended to increase understanding of the overall background of the invention, without answering
It has been the prior art well known to persons skilled in the art when being considered as recognizing or imply that the information is constituted in any form.
Invention content
Goal of the invention
It is active in preparation enhancing normal cell that an object of the present invention is to provide a kind of water-soluble fullerenes derivates
Application in health products.It is normally thin the second object of the present invention is to provide a kind of enhancing comprising the water-soluble fullerenes derivates
The health products of cytoactive.The third object of the present invention, which is to provide a kind of water-soluble fullerenes derivates, enhances normal cell preparing
Application in active cosmetics.The fourth object of the present invention is to provide a kind of enhancing including the water-soluble fullerenes derivates
The active cosmetics of normal cell.
Solution
Purpose to realize the present invention, it is normal in preparation enhancing that the embodiment of the present invention provides a kind of water-soluble fullerenes derivates
Application in the health products of cell activity, wherein the water-soluble fullerenes derivates include：Amido modified fullerene derives
At least one of the fullerene derivate of object, hydroxyl modified；The enhancing normal cell activity includes：Enhance normal cell
The activity of succinate dehydrogenase in mitochondria reducing substances activity and is accelerated in the proliferation of normal cell in enhancing normal cell
At least one.
Purpose to realize the present invention, the embodiment of the present invention additionally provide a kind of active health products of enhancing normal cell,
Including water-soluble fullerenes derivates, wherein the water-soluble fullerenes derivates include：Amido modified fullerene derivate,
At least one of fullerene derivate of hydroxyl modified；The enhancing normal cell activity includes：Enhance the line of normal cell
The activity of succinate dehydrogenase in plastochondria reducing substances activity and is accelerated in the proliferation of normal cell in enhancing normal cell
It is at least one；Optionally, the health products further include at least one suitable for the carrier of health products, diluent or excipient
Kind.
Purpose to realize the present invention, the embodiment of the present invention, which additionally provides a kind of water-soluble fullerenes derivates, to be enhanced preparing
Application in the active cosmetics of normal cell, wherein the water-soluble fullerenes derivates include：Amido modified fullerene
At least one of the fullerene derivate of derivative, hydroxyl modified；The enhancing normal cell activity includes：Enhancing is normal thin
The activity of succinate dehydrogenase in the mitochondria of born of the same parents reducing substances activity and accelerates the increasing of normal cell in enhancing normal cell
At least one of grow.
Purpose to realize the present invention, the embodiment of the present invention additionally provide a kind of active cosmetics of enhancing normal cell,
Including water-soluble fullerenes derivates, wherein the water-soluble fullerenes derivates include：Amido modified fullerene derivate,
At least one of fullerene derivate of hydroxyl modified；The enhancing normal cell activity includes：Enhance the line of normal cell
The activity of succinate dehydrogenase in plastochondria reducing substances activity and is accelerated in the proliferation of normal cell in enhancing normal cell
It is at least one；Optionally, the cosmetics further include the auxiliary material suitable for cosmetics.
Above-mentioned water-soluble fullerenes derivates are preparing the application in enhancing the active health products of normal cell, above-mentioned health care
The application in enhancing the active cosmetics of normal cell, above-mentioned cosmetics exist preparing for product, above-mentioned water-soluble fullerenes derivates
In a kind of possible realization method, the normal cell includes：People's microglia cell (HM cells) and people source liver cell
At least one of (HL-7702 cells).
Above-mentioned water-soluble fullerenes derivates are preparing the application in enhancing the active health products of normal cell, above-mentioned health care
The application in enhancing the active cosmetics of normal cell, above-mentioned cosmetics exist preparing for product, above-mentioned water-soluble fullerenes derivates
In a kind of possible realization method, the activity of succinate dehydrogenase includes in the mitochondria of the enhancing normal cell：It improves just
The quantity of succinate dehydrogenase in the mitochondria of normal cell enhances Board Lot succinate dehydrogenase in the mitochondria of normal cell
At least one of activity.
Above-mentioned water-soluble fullerenes derivates are preparing the application in enhancing the active health products of normal cell, above-mentioned health care
The application in enhancing the active cosmetics of normal cell, above-mentioned cosmetics exist preparing for product, above-mentioned water-soluble fullerenes derivates
In a kind of possible realization method, reducing substances activity includes in the enhancing normal cell：It improves in normal cell
The quantity of at least one of NADPH, FADH, FMNH, NADH enhance in normal cell in NADPH, FADH, FMNH, NADH
The activity of at least one Board Lot or make in NADPH/NADP, FADH/FAD, FMNH/FMN and NADH/NAD at least one
Kind ratio increases.
Above-mentioned water-soluble fullerenes derivates are preparing the application in enhancing the active health products of normal cell, above-mentioned health care
The application in enhancing the active cosmetics of normal cell, above-mentioned cosmetics exist preparing for product, above-mentioned water-soluble fullerenes derivates
In a kind of possible realization method, the amido modified fullerene derivate includes：C60(EDA)9And C70(EDA)9In extremely
Few one kind, wherein EDA represent ethylenediamine base.
Above-mentioned water-soluble fullerenes derivates are preparing the application in enhancing the active health products of normal cell, above-mentioned health care
The application in enhancing the active cosmetics of normal cell, above-mentioned cosmetics exist preparing for product, above-mentioned water-soluble fullerenes derivates
In a kind of possible realization method, the amido modified fullerene derivate is prepared by method comprising the following steps：It will
Fullerene is reacted with ethylenediamine mixing, is carried out post-processing later and is obtained amido modified fullerene derivate；It is wherein described
Fullerene is C60And C70At least one of, the mass ratio of the fullerene and ethylenediamine is 50:45-100.
Above-mentioned water-soluble fullerenes derivates are preparing the application in enhancing the active health products of normal cell, above-mentioned health care
The application in enhancing the active cosmetics of normal cell, above-mentioned cosmetics exist preparing for product, above-mentioned water-soluble fullerenes derivates
In a kind of possible realization method, the fullerene derivate of the hydroxyl modified includes：C60(O)5(OH)12And C70(O)5(OH)12
At least one of.
Above-mentioned water-soluble fullerenes derivates are preparing the application in enhancing the active health products of normal cell, above-mentioned health care
The application in enhancing the active cosmetics of normal cell, above-mentioned cosmetics exist preparing for product, above-mentioned water-soluble fullerenes derivates
In a kind of possible realization method, the fullerene derivate of the hydroxyl modified is prepared by method comprising the following steps：It will
Fullerene is mixed with hydrogenperoxide steam generator and aqueous slkali and is reacted under conditions of 50~80 DEG C；The wherein described fullerene is C60
And C70At least one of, the aqueous slkali is at least one of sodium hydrate aqueous solution and potassium hydroxide aqueous solution.
Above-mentioned water-soluble fullerenes derivates are preparing the application in enhancing the active health products of normal cell, above-mentioned health care
The application in enhancing the active cosmetics of normal cell, above-mentioned cosmetics exist preparing for product, above-mentioned water-soluble fullerenes derivates
In a kind of possible realization method, based on the gross mass of the aqueous slkali, the content of alkali is 8~50wt% in the aqueous slkali；
Based on the gross mass of the hydrogenperoxide steam generator, the content of hydrogen peroxide is 20~40wt% in the hydrogenperoxide steam generator；Institute
It is 30~200mg that fullerene, which is stated, with the mass volume ratio of hydrogenperoxide steam generator and aqueous slkali：5~10ml：1~4ml.
Health products or above-mentioned health products in above application in another embodiment, can be tablets, pill, dissipate
Agent, pastille, sachet, cachet, elixir, suspending agent, emulsion, solution, syrup, aerosol, ointment, soft glutoid glue
The preparation of capsule, suppository, aseptic injectable solution or aseptic packaging powder-injection.Active ingredient water-soluble fullerene is derived in the present invention
Object is prepared into health products, make its after being applied to subject can with quick-release, sustained release or sustained release active ingredient, such as：
Active ingredient can be mixed with carrier, diluted or encapsulated in the carrier with carrier.
Health products or above-mentioned health products in above application in another embodiment, suitable for as carrier, excipient
Some examples with diluent include lactose, dextrose, sucrose, sorbierite, mannitol, starch, resin, Arabic gum, phosphoric acid
Calcium, alginate, tragacanth, gelatin, calcium silicates, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, aqueous syrup
(water syrup), methylcellulose, methyl hydroxybenzoate and propyl ester, talcum powder, magnesium stearate and liquid paraffin.
Health products or above-mentioned health products in above application can also also comprise lubrication in another embodiment
The auxiliary agents such as agent, wetting agent, emulsification and suspending agent, preservative, sweetener or corrigent.
Health products or above-mentioned health products in above application in another embodiment, when health products are deposited in liquid form
When, a concentration of 0.01-50mg/mL of the active ingredient water-soluble fullerenes derivates in the health products be optionally
0.01-10mg/mL, 10-20mg/mL, 10-50mg/mL；When the health products in solid form in the presence of, active ingredient is water-soluble
A concentration of 0.01-50mg/g of the property fullerene derivate in the health products or the Halth-care composition be optionally
0.01-10mg/g, 10-20mg/g, 10-50mg/g.
Cosmetics or above-mentioned cosmetics in above application in another embodiment, are suitable for the accessory package of cosmetics
Include foaming agent, foam stabiliser, surfactant, conditioner, wetting agent, flavouring agent, tackifier, thickener, buffer, anti-corrosion
Agent and sun-screening agent etc..
Advantageous effect
(1) present invention uses two class water-soluble fullerenes derivates：Amido modified fullerene derivate, hydroxyl modified
Fullerene derivate, this two fullerenes derivative has excellent water solubility, and molecular particle size is smaller, easily enters cell
It is internal.Wherein, the surface of amido modified fullerene derivate carries positive charge, the surface of the fullerene derivate of hydroxyl modified
It is negatively charged, all there is excellent stability of solution.
(2) by two fullerene derivatives respectively with people's microglia cell (HM cells) and people source liver cell (HL-
7702 cells) both people source normal cells are incubated jointly, pass through Cell Counting Kit-8 (CCK-8) and Alamar
Two kinds of detection reagents of Blue detect the activity of reproducibility enzyme and reducing substances in normal cell in normal cell mitochondria respectively
Activity.Result of study proves that different water-soluble fullerenes derivates show general character and characteristic, such as：General character shows this
Two class water-soluble fullerenes derivates after 24 hours, remain to show even at a low concentration with HM cells and HL-7702 cell incubations
Write the mitochondria activity and cell overall activity of enhancing people's source HM cells and HL-7702 cells；Property list is now with water solubility
The effect of the increase of fullerene derivate concentration, some water-soluble fullerenes derivates shows concentration inhibition, i.e., with dense
The increase of degree, the effect for enhancing mitochondria activity and cell activity declines instead, and the effect of some water-soluble fullerenes is then
It is continuously increased with the increase of concentration.
(3) simultaneously, this two classes water-soluble fullerenes derivates is apparent without occurring under the high concentration of 500 μM (μm ol/L)
Cytotoxicity, it was demonstrated that it can be widely used in biomedical sector in the biological safety of cellular level.
Description of the drawings
According to below with reference to the accompanying drawings becoming to detailed description of illustrative embodiments, other feature of the invention and aspect
It is clear.
One or more embodiments are illustrated by the picture in corresponding attached drawing, these exemplary theorys
The bright restriction not constituted to embodiment.
Fig. 1 is the C in the embodiment of the present invention 1.460-EDA、C70-EDA、C60-OH、C70The surface potential of-OH is (i.e.:zeta
Current potential).
Specific implementation mode
In order to make the object, technical scheme and advantages of the embodiment of the invention clearer, below in conjunction with one or more real
It applies example and corresponding attached drawing technical solution in the embodiment of the present invention carries out clear, complete exemplary illustration.
Obviously, described embodiments are some of the embodiments of the present invention, instead of all the embodiments.Based on the present invention
In embodiment, the every other implementation that those of ordinary skill in the art are obtained without creative efforts
Example, shall fall within the protection scope of the present invention.Unless otherwise explicitly stated, otherwise in the whole instruction and claims
In, the term " include " or its transformations will be understood as be include stated component part, and
It does not exclude other elements or other components.
These embodiments do not constitute the restriction to protection domain.Unless otherwise indicated, any embodiment here need not
It is construed to preferred or advantageous over other embodiments.
In addition, in order to better illustrate the present invention, numerous details is given in following examples.This field
It will be appreciated by the skilled person that without certain details, the present invention can equally be implemented.In some embodiments, for ability
Method, means, element, the condition usually according to normal condition and described in handbook known to field technique personnel or according to manufacture
The experimental method of condition proposed by manufacturer is not described in detail, in order to highlight the purport of the present invention.Material used, reagent
Deng being that conventional can be obtained by commercial sources unless otherwise specified.
Certainly Japanese eastern Renhua subject skill (Shanghai) limited public affairs of CCK-8 detection kits purchase used in the embodiment of the present invention
Department, article No.：CK04；Alamar Blue detection kits are bought from Beijing Su Lanbao Science and Technology Ltd.s, article No.：A7631.
The people source normal cell used in the embodiment of the present invention is HM cells (purchase is in Bei Na biotechnologies company), it is
Third class cell in central nervous system in addition to neuron and astroglia, plays removing in central nervous system
The effects that apoptotic cell and pathogen.
The people source normal cell used in the embodiment of the present invention be HL-7702 cells (purchase in north receive biotechnology public affairs
Department), it is Human normal hepatocyte system, there are extremely similar biological characteristics with the liver cell in tissue.
Embodiment 1, the synthesis of water-soluble fullerenes derivates and its hydration grain size and zeta current potentials characterization
1.1 amido modified fullerene derivate C60- EDA and C70The synthesis of-EDA.
(a) 100mL conical flask with cover is added in 50mL ethylenediamines (analyzing pure, traditional Chinese medicines reagent, density 0.9mg/ml), be added
50mg solid fullerenes C60(purity：99%, Xiamen good fortune is taken in the fresh material Science and Technology Ltd.), magnetic stir bar is added, is used
Magnetic stirrer (temperature for 24 hours：Room temperature, rotating speed：1000r/min), using solvent filter (volume：1L, filter sizes：
200nm, Jin Teng company) reactant is filtered after obtain brown-red solution.The ingredient of solution does not participate in the second two of reaction mainly
Amine and C60(EDA)9。
(b) solution for obtaining (a) step is added in the round-bottomed flask of 250ml, reuses Rotary Evaporators (model：IKA
RV10basic filtrate) is rotarily dried into (temperature completely：70 degrees Celsius, rotating speed：80r/min).By hydrochloric acid (concentration：1mol/L)
It is added in round-bottomed flask, oscillation flask makes the object that is evaporated on its inner wall be dissolved in dilute hydrochloric acid, obtains brownish red clear solution.
(c) solution that (b) step obtains is neutralized to pH test paper and is detected as faintly acid (pH is 5 or so), to ensure excess
Ethylenediamine with chlorination salt form exist, can fully be removed in subsequent dialysis step.Solution is packed into bag filter after neutralizing
(cutoff 3500), which is put into ultra-pure water, dialyses, and the conductivity dialysed to ultra-pure water is less than 1 μ s/cm.
(d) solution after (c) step being dialysed is flowed through equipped with anion exchange resin (Ambersep900 (OH), Yi Nuokai
Reagent Company) chromatographic column, be repeated 3 times.Solution loading bag filter (cutoff 3500) after ion exchange is put into super
It dialyses in pure water, the conductivity dialysed to ultra-pure water is less than 1 μ s/cm, obtains C60(EDA)9, abbreviation C in the application60-EDA.Solution
In clear brownish red.
Using reaction step identical with 1.1 parts and condition, different is to use 50mg C70Solid (purity：
99%, Xiamen good fortune is taken in the fresh material Science and Technology Ltd.) substitute C60, obtain C70(EDA)9, abbreviation C in the application70-EDA。
Other are to the characterization of above structure referring to patent application CN201610514134.4.
The fullerene derivate C of 1.2 hydroxyl modifieds60- OH and C70The synthesis of-OH.
(a) by 7mL mass percentages be 30% hydrogen peroxide (analysis pure, be purchased from traditional Chinese medicines reagent) aqueous solution and 3mL
100mL round-bottomed flasks are added in sodium hydroxide (analyzing pure, traditional Chinese medicines reagent) aqueous solution that mass percentage is 40%, are added
200mg fullerenes C60Solid (purity：99%, Xiamen good fortune is taken in the fresh material Science and Technology Ltd.), add magnetic stir bar (type
Number：B200), using magnetic stirrer (temperature for 24 hours：70 DEG C, rotating speed：1000r/min), using solvent filter (volume：
1L, filter sizes：200nm, Jin Teng company) brown yellow solution is obtained by filtration.
(b) solution for obtaining (a) step is added in the centrifuge tube of 50ml, adds excessive a concentration of 95% ethyl alcohol
(analyzing pure, traditional Chinese medicines reagent).By centrifugation (rotating speed：10000r/min, time：Upper layer colourless solution 4min) is removed afterwards, will be received
The precipitation of collection is dissolved in ultra-pure water, obtains yellow clear solution.
(c) it the solution that (b) step obtains is fitted into bag filter (cutoff 3500) is put into ultra-pure water and dialyse, thoroughly
It analyses to the conductivity of ultra-pure water and is less than 1 μ s/cm, obtain yellow solution.
(d) solution after (c) step being dialysed is fitted into 50mL plastic centrifuge tubes, dry using freezing is put into after liquid nitrogen frozen
(temperature is freeze-dried in dry machine：- 29 DEG C, vacuum degree：55Pa, time：48h), the yellow solid obtained, i.e. C60(O)5(OH)12,
Abbreviation C in the application60-OH。
Using reaction step identical with 1.2 parts and condition, different be using etc. quality C70Substitute C60,
Obtain C70(O)5(OH)12, abbreviation C in the application70-OH。
Other are to the characterization of above structure referring to patent application 201610515403.9
1.3C60-EDA、C70-EDA、C60-OH、C70The hydration grain size of-OH is tested.
By above-mentioned water-soluble fullerenes derivates be configured to a concentration of 10 μM of aqueous solutions (through repeated detection, aqueous solution it is dense
The influence very little to being hydrated grain size is spent, a concentration of 10 μM is selected and is detected), pass through filter membrane (cut-off grain size is 0.22 μm) filtering
Afterwards, it takes 1 milliliter of solution to be added in the plastic sample pond of four sides light transmission, uses dynamic light scattering (DLS, Nano-ZS90, English
Malvern Co., Ltd of state) test respectively its be hydrated grain size size.Test result is as shown in table 1, C60-EDA、C70-EDA、
C60-OH、C70The hydration grain size of-OH is all mainly distributed on 150nm hereinafter, smaller easy swallowed of its grain size enters cell interior.
The hydration particle diameter distribution of 1 water-soluble fullerenes derivates of table
1.4 C60-EDA、C70-EDA、C60-OH、C70The zeta potential tests of-OH.
Above-mentioned water-soluble fullerenes derivates are configured to a concentration of 10 μM of aqueous solutions, (ending grain size is by filter membrane
0.22 μm) after filtering, is taken in 1 milliliter of solution injection special test pond using syringe, then use potentiometric analyzer (Nano-
ZS90, Malvern Co., Ltd of Britain) test the size of its zeta current potential.Test results are shown in figure 1, C60-EDA、C70-
EDA、C60-OH、C70The zeta Potential distributions of-OH concentrate on 25mV, 40mV, -23mV and -28mV.The symbol of Zeta potential can
Reflect the type of fullerene derivate institute belt surface charge, the bigger expression fullerene derivate stability of solution of absolute value is more
It is good.So as to prove C60- EDA and C70The surfaces-EDA are positively charged, and C60-OH、C70The surfaces-OH are negatively charged, they
Zeta current potential absolute values are all larger, it was demonstrated that it is with preferable stability of solution.
Embodiment 2, water-soluble fullerenes derivates enhance the active experiment of normal cell
2.1 CCK-8 detect normal cell activity
CCK-8 cytoactive detection reagents are 2- (2- methoxyl group -4- nitrobenzophenones) -3- (4- nitrobenzophenones) -5- (2,4-
Disulfonic acid benzene) -2H- tetrazolium monosodium salts].CCK-8 is reduced to by the amber dehydrogenase in cell mitochondrial with high water soluble
Yellow first a ceremonial jade-ladle, used in libation product (Formazan), the quantity of the first a ceremonial jade-ladle, used in libation object of generation is directly proportional to the quantity of living cells, is examined with enzyme linked immunological
Survey instrument and measure its absorbance value at 450nm wavelength, by detect reproducibility enzyme in cell mitochondrial it is active by between it is reversed
Reflect cell activity.
Using containing 10% N of embryo's serum low sugar DMEM culture mediums and RPMI-1640 culture mediums respectively to HM cells and
HL-7702 cells carry out incubation passage；HM cells and the cell suspension of HL-7702 are obtained using trypsin digestion, with 5 ×
104The cell density in a/hole is inoculated in 96 transparent well culture plates, and 200 μ L cell suspensions (HM cells or HL- are added per hole
7702 cells), in 37 DEG C and 5%CO2Cell incubator in culture 24 hours, cell is attached to 96 orifice plate bottom growns；It is several
Experimental group removes upper layer culture medium, is separately added into 200 μ L and contains a concentration of 2.5,5,10,20,40,50,100,200,500 μM
C60-EDA、C70-EDA、C60-OH、C70The fresh culture of-OH, every group of 6 multiple holes, and blank control group only needs to change into fresh training
Base is supported, 96 well culture plates are placed in cell incubator and are incubated 24 hours；Upper layer culture medium is removed, the colourless trainings of 90 μ L are changed to
Support base and 10 μ L CCK-8, be incubated 60 minutes to culture medium color by it is orange red become glassy yellow when, with microplate reader in 450nm
The absorbance in each hole, by specification require to calculate its opposite cell activity, HM cells and HL-7702 on 96 orifice plates of place's test
The testing result of cell is shown in Table 2 and table 3 respectively.
Raising ratio of the fullerene derivate of 2 various concentration of table to HM cell activity
As shown in table 2, for HM cells, the fullerene derivate of amido modified fullerene derivate and hydroxyl modified exists
Amber dehydrogenase activity in HM cell mitochondrials can be effectively improved under 2.5 μM of low concentration.In 2.5-500 μM of concentration range
Interior, amber dehydrogenase activity is with C in HM cell mitochondrials60-EDA、C70- EDA or C60The concentration of-OH increases and improves, and has bright
Aobvious dose-effect relationship, but when improving the characteristics of, is different.C60-EDA、C70- EDA in 2.5-40 μM of concentration range, activity improve
Speed will be slower than C60- OH, but the two can continue to increase active humidification, in 500 μM of high concentration, to active enhancing
Effect is maximum；And C60- OH in 2.5-40 μM of concentration range, activity improve speed faster, its increased activity at 40 μM
When percentage is 2.5 μM more than 8 times, but in 40-500 μM of concentration range, increased activity acts on unobvious.And C70- OH is then
Peak value is reached to increased activity effect in a certain concentration, is then begun to decline later.Amido modified fullerene derivate C60-EDA
Can amber dehydrogenase activity activity significantly more be improved 40%.
Enhancing effect of the fullerene derivate of 3 various concentration of table to HL-7702 cell activity
As shown in table 3, for HL-7702 cells, the fullerene of amido modified fullerene derivate and hydroxyl modified spreads out
For biology under 40 μM of concentration, cell activity can be improved 20% or more by water-soluble fullerenes derivates.But C60-EDA、C60-
OH、C70- OH is the decline for starting to occur increased activity effect after reaching a certain concentration, only C70- EDA is at 100-500 μM
In the range of, amber Dehydrogenase activtity performance in HL-7702 cell mitochondrials is continued to enhance, the enhancing for being finally reached 43% is made
With.Illustrate that different water-soluble fullerenes is different to the enhancing effect of HL-7702 cells.
Above-mentioned experimental result can prove the fullerene derivate energy of amido modified fullerene derivate and hydroxyl modified
Enough significantly increase amber dehydrogenase activity in the cell mitochondrial of HM and HL-7702, and amber in the mitochondria of CCK-8 detections
Dehydrogenase activity can reflect the activity of cell.In addition, under 500 μM of high concentration, the cell mitochondrial of all experimental groups
Middle amber Dehydrogenase activtity activity is all higher than blank control group, illustrates that this three classes fullerene derivate does not have apparent cytotoxicity.
Mitochondria is the energy plants of cell, and cyclophorase is mainly activated cell self-energy metabolism, is cell, group
It knits and provides energy with the normal physiological activity of organism；Mitochondria enzyme activity can reflect that its is new at metabolic capability, and enzymatic activity increases
It is strong to also mean that mitochondria has higher energy supply efficiency, for maintain normal physiological function and resist exogenous damage and
Stimulation is all of great significance.
2.2 Alamar Blue detection normal cell activity
Alamar blue cytoactive detections reagents are changed into a reduction state also to be originated in pink or red fluorescence
Object, for excitation wavelength between 530-560nm, the preferred excitation wavelength of the present invention is 550nm, wavelength of transmitted light 590nm.
In proliferation process, intracellular NADPH/NADP, FADH/FAD, the ratio raising of FMNH/FMN and NADH/NAD, intake is carefully
The Alamar blue of intracellular are discharged into extracellular and are dissolved in culture medium after these Metabolic Intermediates and cytochromes reduction
In, it is detected by fluophotometer, fluorescence intensity can reflect the general activity state of cell.
NADPH, NADP, FADH, FAD, FMNH, FMN, NADH, NAD are intracellular important coenzyme, be involved in sugar,
Most redox reactions of fat, three big metabolism of protein play important work during glycometabolism generates ATP
With；Ratio appropriate, which increases, means that intracellular whole metabolic capability improves, and intracellular reduced level improves, has more
Strong anti-oxidative damage ability allows living organism to have stronger vigor and applicable ability.
Using containing 10% N of embryo's serum low sugar DMEM culture mediums and RPMI-1640 culture mediums respectively to HM cells and
HL-7702 cells carry out incubation passage；HM cells and the cell suspension of HL-7702 are obtained using trypsin digestion, with 5 ×
104The cell density in a/hole is inoculated in 96 well culture plates of black, and 200 μ L cell suspensions (HM cells or HL- are added per hole
7702 cells), in 37 DEG C and 5%CO2Cell incubator in culture 24 hours, cell is attached to 96 orifice plate bottom growns；It removes
Upper layer culture medium, each experimental group are separately added into the C that 200 μ L contain a concentration of 2.5,5,10,20,40 μM60-EDA、C70-EDA、
C60-OH、C70The fresh culture of-OH, every group of 6 multiple holes, and to be added to the fresh culture of phosphate buffer solution (PBS)
As blank control group, experimental group and blank control group are placed in cell incubator and are incubated 24 hours；Upper layer culture medium is removed,
It is changed to the colourless culture mediums of 90 μ L and the Alamar blue (concentration of 10 μ L：0.01mg/ml), 60 minutes are incubated to culture medium
When color becomes pink by purple, 96 are tested at 550nm excitation wavelengths and 590nm Detection wavelengths using multi-function microplate reader
The fluorescence intensity in each hole on orifice plate indicates its opposite cell activity with specification requirement.
Enhancing effect of the fullerene derivate of 4 various concentration of table to HM cell activity
Alamar Blue are the activity of the intracellular reducing substances for participating in metabolism of detection, can reflect cell from side
Activity.As shown in table 4, for HM cells, C60-EDA、C70-EDA、C60- OH is under 2.5 μM of low concentration to cell activity
Enhancing effect unobvious can only significantly improve cell activity under 10 μM or more of high concentration.C70- OH is at 2.5 μM
10% can be reached to the enhancing effect of cell activity under low concentration, but improve C70After the concentration of-OH, the enhancing to cell activity
Effect does not significantly increase.
Enhancing effect of the fullerene derivate of 5 various concentration of table to HL-7702 cell activity
As shown in table 5, for HL-7702 cells, C60-EDA、C70-EDA、C60-OH、C70- OH can be under 2.5 μM of concentration
Effectively enhance cell activity, it is more notable to the enhancing effect of cell activity under higher concentration.
In short, C60-EDA、C70-EDA、C60-OH、C70- OH can significantly increase the mitochondria activity of human archeocyte
(CCK-8 detections), while more significantly improving the activity (Alamar for the reducing substances for being related to metabolism in human archeocyte
Blue is detected).Meanwhile C60-EDA、C70-EDA、C60-OH、C70- OH is apparent thin without occurring under 500 μM of high concentration
Cellular toxicity, it was demonstrated that this three classes water-soluble fullerenes derivates all has significant biological safety on a cellular level, can be by
It is widely used in biomedical sector.
Embodiment 3, water-soluble fullerenes derivates accelerate the proliferation of normal cell
Using -2 deoxyuridine of 5- acetenyls (EDU) and flow cytomery C60- EDA, C70- EDA, C60-
OH, C70After several water-soluble fullerenes derivates of-OH are incubated HM cells and HL-7702 cells for 24 hours, the shadow of cell proliferation generation
It rings.Detection method is as follows：
1, the preparation of fluorescent dye：
A, the preparation of the EDU solution of 10mM：The dimethyl sulfoxide (DMSO) of 4mL is taken to be added in the EDU reagent bottles of 10mg, with 1mL's
Liquid-transfering gun piping and druming is uniform, is then placed in storing liquid in -20 DEG C of refrigerator and preserves, and reagent can save 1 year；
B, the preparation of 488 azide of Alexa Fluor：Take the dimethyl sulfoxide (DMSO) of 130 μ L that Alexa Fluor are added
In the reagent bottle of 488 azide, is blown and beaten uniformly with the liquid-transfering gun of 200 μ L, then storing liquid is placed in -20 DEG C of refrigerator and is protected
It deposits, reagent can save 1 year；
C, the preparation of 1% bovine serum albumin：Culture bottle of the bovine serum albumin in 100mL of 1g is weighed using assay balance
In, the PBS of the 1X of 100mL is added, is uniformly mixed, then storing liquid is placed in 4-8 DEG C of refrigerator and is preserved；
D, the preparation of 1X Click-iT penetrating fluids and washing solution：It takes the 1X Click-iT penetrating fluids of 5mL and washs molten
Liquid is added in the culture bottle of 50mL in 1% bovine serum albumin to PBS buffer solutions, is uniformly mixed, then sets storing liquid
It is preserved in 4-8 DEG C of refrigerator；
E, the preparation of the Click-IT EDU buffer additives of 10X：The ultra-pure water of 2mL is taken to be placed in the Click-IT of 10X
It in the reagent bottle of EDU buffer additives, is uniformly mixed, then storing liquid is placed in -20 DEG C of refrigerator and is preserved, reagent can save
1 year；
2, specific steps：
A, EDU marks cell：
Using containing 10% N of embryo's serum low sugar DMEM culture mediums and RPMI-1640 culture mediums respectively to HM cells and
HL-7702 cells carry out incubation passage；Cell is prepared into after HM cells or HL-7702 cells are digested with pancreatin with culture medium to hang
Liquid, with 1 × 105Cell density by cell kind in 96 orifice plates, each cell for each water-soluble fullerenes derivates (such as
HM cells are directed to C60- EDA) it is 1 group, every group is planted three holes 1 respectively, and 2,3, wherein：1 hole is blank control wells, and 2 holes are negative right
According to hole, 3 holes are experimental port, are arranged altogether 8 groups (2 kinds of cells, 4 kinds of water-soluble fullerenes).In every group：The thin of 2mL is added in each hole
Born of the same parents' suspension (HM cells or HL-7702 cells), after being incubated for 24 hours, culture medium is replaced in 1, No. 2 hole, and No. 3 hole additions are prepared with culture medium
Final concentration of 40 μM of correspondence water-soluble fullerene (i.e.：C60-EDA、C70-EDA、C60- OH or C70- OH) it is incubated for 24 hours, then
The culture solution for removing 2,3 holes, is then washed twice with the PBS of 1X, the final concentration then prepared with culture medium in 2, No. 3 hole additions
For 10 μM of EDU dye solutions (note：Original liquid concentration is 10mM, therefore first takes the EDU stostes of 100 μ L, then add 900 μ L it is ultrapure
Water dilutes, and final concentration of 10 μM are diluted to culture medium again after), it is incubated 3h.
B, cell is fixed and is permeated
The cell in 2, No. 3 holes is digested with pancreatin, the cell suspension that 1mL is prepared into culture solution is placed in 10mL centrifuge tubes,
Supernatant is removed in centrifugation, and 1% bovine serum albumin that 3mL is then added washes twice, and removes supernatant, in each centrifuge tube
The Click-iT fixers of 100 μ L are added, are blown and beaten uniformly with the liquid-transfering gun of 200 μ L, room temperature is incubated 15min under the conditions of being protected from light, so
1% bovine serum albumin that 3mL is added afterwards washes twice, and removes supernatant, the 1X of 100 μ L is added in each centrifuge tube
Click-It reactive infiltrations liquid and cleaning solution are blown and beaten uniformly with the liquid-transfering gun of 200 μ L, and room temperature is incubated 15min under the conditions of being protected from light, so
1% BSA that 3mL is added afterwards is washed twice；
C, Click-iT reacts：
The preparation of the Click-iT EDU buffer additives of 1X：By 1:10 ratio dilutes the Click- of 10X with ultra-pure water
IT EDU buffer additives；
Click-iT Reation cocktails
It prepares Click-iT reaction solutions and can refer to following table
The Click-iT Reation cocktails of 500 μ L are added in each centrifuge tube of step B, are uniformly mixed,
Room temperature is incubated 15min under the conditions of being protected from light, the 1X Click-iT reactive infiltrations liquid of 3mL then is added and cleaning solution washed once, goes
Fall supernatant, often the 1X Click-It reactive infiltrations liquid of 1mL is added in pipe and cleaning solution prepares cell suspension；Then No. 1 hole is used
Pancreatin digests, and with the cell suspension for being prepared into 1mL of 1X Click-It reactive infiltrations liquid and cleaning solution, then up flow type detects,
Its excitation wavelength 488nm's, launch wavelength：530-560nm.
Experimental result is shown：
There is division growth (by the way that " the S phase cells of DNA replication dna are total after cellular control unit is incubated for 24 hours for HM cells
Accounting in cell " indicates), the daughter cell of the S phases cell and division completion that are in DNA replication dna accounts for the 30.66% of cell total amount；
Under the same conditions, a concentration of 40 μM of C60-EDA、C70-EDA、C60-OH、C70After-OH is incubated HM cells for 24 hours, it is in DNA
The daughter cell accounting that the S phases cell of duplication and division are completed is respectively 40.69%, 44.26%, 39.11%, 44.68%, thin
Born of the same parents' opposite proliferation rate has been respectively increased 10.03%, 13.60%, 8.45%, 14.02%；
For HL-7702 cells, the S phases cell for being in DNA replication dna of control group and the daughter cell of division completion account for cell
Total amount is 21.53%, a concentration of 40 μM of C60-EDA、C70-EDA、C60-OH、C70After-OH is incubated for 24 hours, it is in DNA replication dna
The ratio that the daughter cell that S phases cell and division are completed accounts for cell total amount is respectively increased to 34.76%, 34.86%, 31.85%,
36.90%, cell proliferation rate has increased separately 13.23%, 13.33%, 10.32%, 15.37%.
To prove that water-soluble fullerenes derivates can accelerate HM cells and HL-7702 cell Proliferations, by multiplication rate
10% or so is improved, without result in malignant proliferation.
Finally it should be noted that：The above embodiments are merely illustrative of the technical solutions of the present invention, rather than its limitations；Although
Present invention has been described in detail with reference to the aforementioned embodiments, it will be understood by those of ordinary skill in the art that：It still may be used
With technical scheme described in the above embodiments is modified or equivalent replacement of some of the technical features；
And these modifications or replacements, various embodiments of the present invention technical solution that it does not separate the essence of the corresponding technical solution spirit and
Range.
Claims (10)
1. a kind of water-soluble fullerenes derivates are preparing the application in enhancing the active health products of normal cell, wherein the water
Dissolubility fullerene derivate includes：At least one in the fullerene derivate of amido modified fullerene derivate, hydroxyl modified
Kind；The enhancing normal cell activity includes：Enhance the activity of succinate dehydrogenase in the mitochondria of normal cell, enhancing normally
Intracellular reducing substances activity and at least one of the proliferation for accelerating normal cell.
2. a kind of active health products of enhancing normal cell comprising water-soluble fullerenes derivates, wherein described water-soluble rich
Strangling ene derivative includes：At least one of the fullerene derivate of amido modified fullerene derivate, hydroxyl modified；It is described
Enhancing normal cell activity includes：Enhance the activity of succinate dehydrogenase in the mitochondria of normal cell, in enhancing normal cell
Reducing substances activity and at least one of the proliferation for accelerating normal cell；Optionally, the health products further include being suitable for
At least one of carrier, diluent or excipient of health products.
3. a kind of water-soluble fullerenes derivates are preparing the application in enhancing the active cosmetics of normal cell, wherein the water
Dissolubility fullerene derivate includes：At least one in the fullerene derivate of amido modified fullerene derivate, hydroxyl modified
Kind；The enhancing normal cell activity includes：Enhance the activity of succinate dehydrogenase in the mitochondria of normal cell, enhancing normally
Intracellular reducing substances activity and at least one of the proliferation for accelerating normal cell.
4. a kind of active cosmetics of enhancing normal cell comprising water-soluble fullerenes derivates, wherein described water-soluble rich
Strangling ene derivative includes：At least one of the fullerene derivate of amido modified fullerene derivate, hydroxyl modified；It is described
Enhancing normal cell activity includes：Enhance the activity of succinate dehydrogenase in the mitochondria of normal cell, in enhancing normal cell
Reducing substances activity and at least one of the proliferation for accelerating normal cell；Optionally, the cosmetics further include being suitable for
The auxiliary material of cosmetics.
5. water-soluble fullerenes derivates according to claim 1 are in preparing the enhancing active health products of normal cell
Using, health products according to claim 2, water-soluble fullerenes derivates according to claim 3 prepare increase
Application in the active cosmetics of strong normal cell or cosmetics according to claim 4, it is characterised in that：It is described normal
Cell includes：At least one of people's microglia cell and people source liver cell.
6. water-soluble fullerenes derivates according to claim 1 are in preparing the enhancing active health products of normal cell
Using, health products according to claim 2, water-soluble fullerenes derivates according to claim 3 prepare increase
Application in the active cosmetics of strong normal cell or cosmetics according to claim 4, it is characterised in that：The enhancing
The activity of succinate dehydrogenase includes in the mitochondria of normal cell：Improve succinate dehydrogenase in the mitochondria of normal cell
Quantity, at least one of the activity for enhancing Board Lot succinate dehydrogenase in the mitochondria of normal cell.
7. water-soluble fullerenes derivates according to claim 1 are in preparing the enhancing active health products of normal cell
Using, health products according to claim 2, water-soluble fullerenes derivates according to claim 3 prepare increase
Application in the active cosmetics of strong normal cell or cosmetics according to claim 4, it is characterised in that：The enhancing
Reducing substances activity includes in normal cell：Improve at least one of NADPH, FADH, FMNH, NADH in normal cell
Quantity enhances the activity of the Board Lot of at least one of NADPH, FADH, FMNH, NADH in normal cell or makes NADPH/
At least one of NADP, FADH/FAD, FMNH/FMN and NADH/NAD ratio increase.
8. water-soluble fullerenes derivates according to claim 1 are in preparing the enhancing active health products of normal cell
Using, health products according to claim 2, water-soluble fullerenes derivates according to claim 3 prepare increase
Application in the active cosmetics of strong normal cell or cosmetics according to claim 4, it is characterised in that：The amino
The fullerene derivate of modification includes：C60(EDA)9And C70(EDA)9At least one of, wherein EDA represents ethylenediamine base；Institute
The fullerene derivate for stating hydroxyl modified includes：C60(O)5(OH)12And C70(O)5(OH)12At least one of.
9. water-soluble fullerenes derivates according to claim 1 are in preparing the enhancing active health products of normal cell
Using, health products according to claim 2, water-soluble fullerenes derivates according to claim 3 prepare increase
Application in the active cosmetics of strong normal cell or cosmetics according to claim 4, it is characterised in that：The amino
The fullerene derivate of modification is prepared by method comprising the following steps：Fullerene is mixed with ethylenediamine and is reacted, it
Post-processing is carried out afterwards obtains amido modified fullerene derivate；The wherein described fullerene is C60And C70At least one of, institute
The mass ratio for stating fullerene and ethylenediamine is 50:45-100.
10. water-soluble fullerenes derivates according to claim 1 are in preparing the enhancing active health products of normal cell
Application, it is prepared by health products according to claim 2, water-soluble fullerenes derivates according to claim 3
Enhance the application in the active cosmetics of normal cell or cosmetics according to claim 4, it is characterised in that：The hydroxyl
The fullerene derivate of base modification is prepared by method comprising the following steps：By fullerene and hydrogenperoxide steam generator and alkali soluble
Liquid is mixed and is reacted under conditions of 50~80 DEG C；The wherein described fullerene is C60And C70At least one of, the aqueous slkali
For at least one of sodium hydrate aqueous solution and potassium hydroxide aqueous solution；Optionally, the gross mass based on the aqueous slkali, institute
The content for stating alkali in aqueous slkali is 8~50wt%；Optionally, the gross mass based on the hydrogenperoxide steam generator, the peroxidating
The content of hydrogen peroxide is 20~40wt% in hydrogen solution；Optionally, the fullerene and hydrogenperoxide steam generator and aqueous slkali
Mass volume ratio be 30~200mg：5~10ml：1~4ml.
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| CN108888534A (en) * | 2018-08-16 | 2018-11-27 | 佛山市洵腾科技有限公司 | A kind of anti-wrinkle eye cream and preparation method thereof |
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| CN112516164A (en) * | 2020-12-01 | 2021-03-19 | 中国科学院化学研究所 | Amino fullerene material for inhibiting tumor metastasis |
| CN114099366A (en) * | 2021-11-17 | 2022-03-01 | 珠海安肽生物医药技术开发有限公司 | Sterile anti-aging wrinkle-removing combined liquid for promoting cell energy metabolism and preparation method and application thereof |
| CN114099366B (en) * | 2021-11-17 | 2023-06-20 | 珠海安肽生物医药技术开发有限公司 | Sterile anti-aging wrinkle-removing combined liquid for promoting cell energy metabolism and preparation method and application thereof |
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Application publication date: 20180727 |