CN106474064B - Artemether nanoliposome and preparation method and application thereof - Google Patents
Artemether nanoliposome and preparation method and application thereof Download PDFInfo
- Publication number
- CN106474064B CN106474064B CN201510533475.1A CN201510533475A CN106474064B CN 106474064 B CN106474064 B CN 106474064B CN 201510533475 A CN201510533475 A CN 201510533475A CN 106474064 B CN106474064 B CN 106474064B
- Authority
- CN
- China
- Prior art keywords
- artemether
- liposome
- nanoliposome
- ultrasonic
- preparation
- 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.)
- Active
Links
Landscapes
- Medicinal Preparation (AREA)
Abstract
The invention discloses an artemether nanoliposome and a preparation method and application thereof. The artemether nanoliposome is prepared from the following raw materials in parts by weight: 1 part of artemether, 2 parts of cholesterol and 8-10 parts of egg yolk lecithin. The preparation method comprises a film dispersion-ultrasonic-film filtration method and an ethanol injection-ultrasonic method. The application of the artemether nano-liposome is the application of the artemether nano-liposome in preparing artemether targeted preparation medicaments. The encapsulating rate of the artemether nanoliposome is 61.33-63.86%, the average particle size is 161.65-162.73 nm, and the dispersion index PDI is 0.2678-0.4463. The artemether nanoliposome provided by the invention has good storage stability at 4 ℃.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to an artemether nanoliposome and a preparation method and application thereof.
Background
Artemether is a derivative of artemisinin and is mainly obtained by semi-synthesis of artemisinin. The artemisinin is an effective component extracted and separated from Artemisia annua (Artemisia) of Compositae family herbaceous distributed in Guangxi, Yunnan, Sichuan and other places in China, and the industrially synthesized artemether has mature technology and large synthesis amount.
The antimalarial effect and effect of artemisinin are internationally acknowledged, so far, artemisinin derivatives have been used as antimalarial derivatives, and mainly include dihydroartemisinin (dihydroartemesinin), arteether, Artesunate (Artesunate) and artemether derivatives, wherein the antimalarial effect of artemether is superior to other artemisinin derivatives.
In addition to its use in the treatment of malaria, artemisinin and its derivatives have a broader range of biological activity. In recent years, the potential application of the medicine also comprises the treatment of schistosomiasis, pneumocystis carinii, toxoplasma, human cytomegalovirus, herpes simplex virus, hepatitis B and hepatitis C virus infection.
Modern research on artemether shows that the artemether has an anti-brain tumor effect, and wuzhiping et al conducts experimental research on the anti-tumor effect of artemether on SD rat brain in-situ inoculated C6 brain glioma according to the characteristics that artemisinin and derivatives thereof can selectively kill tumor cells, have low toxicity on normal tissue cells, and can penetrate through a blood-brain barrier (blood-brain barrier) to enter brain tissues. Experimental results show that the oral artemether has obvious inhibition effect on the brain glioma of SD rats inoculated with C6 brain in situ, and the inhibition effect is enhanced along with the increase of the dosage. The result of experiments on glioma by Deng Xingli et al shows that artemether can inhibit the growth of C6 glioma cells, and the inhibition effect of artemether is time-dependent and concentration-dependent; artemether interferes with the cell cycle of C6 glioma cells, blocks them in the G0-G1 phase and induces their apoptosis.
The research on the anti-tumor effect and the action mechanism of the artemether and cisplatin combined use on L ewis lung cancer of mice are studied by wanghuang and the like, the experimental result shows that the artemether and cisplatin combined use has additive inhibition effect on L ewis lung cancer of mice, can promote tumor cell apoptosis, and the mechanism may be related to the regulation and control of the expression of genes related to apoptosis, the Wanyan and the like investigate the cytotoxic effect of the artemisinin drugs on colon adenocarcinoma cells L S174T, the thiazole blue (MTT) method is adopted to detect the inhibition conditions of drugs with different concentrations and on cell proliferation after acting for different times respectively, and calculate the inhibition rate and calculate the half-number inhibition concentration (IC 50), wherein the inhibition effect of the artemisinin drugs on colon adenocarcinoma cells L S174T proliferation is better, the concentration dependence and time dependence of artemisinin on the time of the drugs can not only be increased when the anti-tumor effect of the artemether and the residence time of the artemisinin drugs on the CNE is better than the survival rate of other tumors of the CNE-1-tumor-resistant cells when the lipid-artemether-based on the lung cancer cell proliferation is studied by the research, the research on the high bioavailability of the high-cholesterol-lipid-based on the high-lipid-.
The concept of targeted agents was proposed by Ehrlich in 1906. The target preparation is also called Target Drug Delivery System (TDDS), and is a novel pharmaceutical preparation technology and process capable of purposefully delivering drugs to focus parts so as to reduce the dosage of the drugs and facilitate control of the speed and mode of drug delivery. The brain targeting preparation targets (i.e. focus position) at brain, and the medicine can penetrate Blood Brain Barrier (BBB) to treat diseases.
There are two categories of targeted agents: the first is the biophysical targeting formulation, the biochemical targeting formulation, the bioimmunological targeting formulation, the dual and multiple targeting formulations, in which the carrier penetrates the tissue target site. Secondly, the active targeting preparation, the passive targeting preparation and the precursor targeting medicine are divided according to the type of the targeting source power, wherein the passive targeting preparation is more researched, and the preparation mainly represents nano-capsule preparation, microspheres and liposome. In this study, brain-targeted nanoliposomes belong to the group of passive targeting agents.
With the development of science and technology and the improvement of technology, the expected drug effect which is originally impossible to achieve reaches the expected effect, and the requirements on auxiliary materials and safety thereof are higher and higher due to deeper understanding of the medicine, the liposome is used as a new inclusion technology, has better affinity with organisms, can reach a focus part, and has small toxic and side effects, the Liu-lan and the like take lecithin and an amphiphilic material P L-GA-PEG as liposome materials, meanwhile, a targeting group (angiopep) is introduced on the surface of the liposome, the taxol nanoliposome is prepared by using a nano precipitation method, the uptake condition of the taxol liposome by brain glioma cells U87MG is detected by a Nile red fluorescent probe molecule detection method, then the targeting of the taxol nanoliposome is determined by using MTT (thiazole blue) to determine the influence of the taxol liposome on the proliferation of U87MG cells, the result shows that the prepared taxol liposome is about 100nm in size, and the fluorescence imaging shows that the targeting taxol liposome is easier to be taken by the U87MG cells of the U87MG cells and the yellow to the Hua and the like than the non-targeting taxol liposome[23]Summarizing the common type of brain targeting liposomes, ① surface modified liposomes can form sterically stable liposomes by modifying polyethylene glycol (PEG) chains on the liposome surface, thus avoiding recognition by phagocytes, prolonging the blood circulation time of the liposomes, and increasing their brain targeting property. ② multi-receptor mediated liposomes can enhance nonspecific binding by modifying multiple ligands, or individual targeting molecules can interfere with specific binding to receptors each other. ③ thermosensitive liposomes, which are characterized in that physical transfer of phospholipids in the liposomes from a colloidal state to a liquid crystal state occurs at a phase transition temperature, thus greatly increasing permeability of liposome membranes, and most drugs are released. ④ others (such as magnetic liposomes, etc.) are selectively concentrated at the targeting site by the action of an external magnetic field and can be used for brain targeting drug delivery.
Disclosure of Invention
The first purpose of the invention is to provide an artemether nanoliposome; the second purpose is to provide a preparation method of the artemether nanoliposome; the third purpose is to provide the application of the artemether nanoliposome.
The first purpose of the invention is realized by that the artemether nanoliposome is prepared by the following raw materials by weight: 1 part of artemether, 2 parts of cholesterol and 8-10 parts of egg yolk lecithin.
The second purpose of the invention is realized by adopting any one of the following methods:
(1) thin film dispersion-ultrasonic-membrane filtration method: dissolving artemether, cholesterol and egg yolk lecithin in an organic solvent, performing ultrasonic oscillation to dissolve the artemether, performing rotary evaporation and reduced pressure evaporation to remove the organic solvent after complete dissolution to form a film, adding water for injection to hydrate the film until the film falls off and dissolves, performing ultrasonic treatment for 180s, filtering the film through a 0.45 mu m filter membrane, and filtering the film through a 0.22 mu m filter membrane to obtain the target artemether nanoliposome;
(2) ethanol injection-sonication: dissolving artemether, cholesterol and egg yolk lecithin in an organic solvent, uniformly dissolving by ultrasonic oscillation, obtaining a solution a after complete dissolution, dropwise adding the solution a into a 5% glucose injection containing tween-80, stirring continuously, oscillating and hydrating at 40 ℃ to obtain a suspension b, evaporating the organic solvent in the suspension b by rotary evaporation and reduced pressure to obtain a crude liposome, and carrying out ultrasonic treatment for 120s, and filtering with a 0.22 mu m filter membrane for 2-4 times to obtain the target artemether nanoliposome.
The third purpose of the invention is realized by the application of the artemether nanoliposome in the preparation of artemether targeted preparation drugs.
The encapsulating rate of the artemether nanoliposome is 61.33-63.86%, the average particle size is 161.65-162.73 nm, and the dispersion index PDI is 0.2678-0.4463. The artemether nanoliposome provided by the invention has good storage stability at 4 ℃.
The liposome (liposome) is formed by Bangham, UK[23]Doctor discovered and named in 1965. He and colleagues found in one study that phospholipid dispersion in water can form closed bilayer vesicles, the center and outer layers of which are separated by water, with a structure similar to a biofilm. Pharmacology is defined as the encapsulation of a drug within a lipid bilayerAnd forming the micro-bubble body.
The liposome is a capsule formed by taking egg yolk lecithin, soybean lecithin or other lecithins as lipid materials, and the phospholipids have amphiphilicity, namely hydrophilicity and lipophilicity. When amphipathic phospholipids and sphingolipids are dispersed in an aqueous phase, the hydrophobic tails of the molecules tend to clump together, avoiding the aqueous phase, while the hydrophilic heads are exposed to the aqueous phase, forming closed vesicles with a bilayer structure. Cholesterol and phosphatidic acid are added into the liposome, and the cholesterol is added to increase the rigidity of a phospholipid bilayer and reduce the mobility of phospholipid so as to properly improve the encapsulation efficiency and stability of the liposome; the purpose of increasing phosphatidic acid is to modulate the zeta potential of the liposomes, reducing their aggregation and increasing their storage stability.
The liposome can be divided into common liposome and nano liposome according to the size of the particle size, generally, the particle size range of the common liposome is 1-100 μm, the particle size is larger, the common liposome is difficult to penetrate through a biological membrane, the stability is poor, the common liposome is easy to agglomerate, and the common liposome has certain limitation in practical application. There is no uniform statement in the limit academic world of nanoliposomes, and it is generally considered that the particle size is less than 200nm, and the nanoliposome has targeting property and can penetrate cells, so many researches are made in recent years]。
Liposomes can be classified by structure essentially as small unilamellar liposomes (SUV) containing only a single bilayer membrane, multilamellar liposomes (M L V) containing multiple concentric bilayer membranes, multivesicular liposomes (MV L) containing multiple non-concentric bilayer membranes.
Modern research into liposomes is increasing, and in combination with current disease and functional requirements, the appearance of new liposomes is endless, and there are several:
1) flexible nano liposome (FN L), also called transfersome, belonging to lipoid aggregate, generally with particle size of tens to hundreds of nanometers, mainly comprising lecithin and surfactant (such as cholesterol, sodium cholate, etc.), wherein the surfactant is embedded in liposome membrane to make liposome have high deformability, under the influence of external force, the time of flexible liposome and pure water with same volume passing through the polymer membrane with pore size smaller than several times of its own diameter is almost the same, while the time of common liposome passing through the polymer membrane is obviously prolonged, so the flexible nano liposome is widely used as carrier of various macromolecules, water-soluble drugs or fat-soluble drugs.
2) Magnetic nano-liposome: the magnetic targeting drug delivery system is prepared from drugs, appropriate magnetic materials and necessary auxiliary materials, and can selectively reach and be positioned at a tumor target area under the guidance of an adequate in vitro magnetic field and along with the operation of blood flow. The drug is released from the carrier in a controlled manner and then exerts its pharmacological effect at the cellular or subcellular level of the tumor tissue, thus having no great influence on normal tissues]. The xylonite and the like prepare the nano liposome with good in-vitro magnetic field responsiveness, uniform particle size distribution, superparamagnetism and ferromagnetism, and the particle size of the magnetic nano liposome is 134.3 nm.
3) Targeting long-circulating nanoliposome: the liposome surface is modified, so that the properties and biological characteristics of the liposome can be effectively controlled, for example, the PEG coating is carried out on the liposome, the recognition of a reticuloendothelial system can be avoided, the circulation time of the liposome in vivo is prolonged, or the liposome surface is connected with a specific ligand (such as an antibody, a polypeptide, a hormone and a sugar), and the target cell can be specifically targeted, so that the active targeting is realized. The liver cancer targeted taxol long-circulating nano liposome prepared by the Thangyong and the like can effectively improve the distribution of taxol medicaments in vivo and improve the safety and curative effect of the taxol medicaments in vivo.
4) Cationic liposome: the cationic liposome generally comprises a cationic amphiphilic compound such as cholesterol derivative or Dimethyl Dioctadecyl Ammonium Bromide (DDAB) and a neutral lipid such as Phosphatidylcholine (PC) or dioleoyl phosphatidylethanolamine (DOPE), has good biocompatibility, can transfer large-charge molecules such as DNA, RNA, ribosome and the like and biological macromolecules, and is an excellent non-viral gene vector.
5) Proliposome: adding one or more cryoprotectants (such as sucrose, glucose, mannitol, etc.) into liposome suspension, and freeze drying the suspension to obtain powder or granule, i.e. precursor liposome, which solves the problems of influence of high temperature sterilization and poor stability of liposome suspension[25]。
6) And others: the novel liposomes such as temperature sensitive liposome, photosensitive liposome, pH sensitive liposome, immunoliposome and the like come into wide range, which adapt to the social demand and bring hope to people suffering from diseases.
① is biodegradable in host body, and has no toxicity or toxic side effect, ② has similarity and histocompatibility with biological membrane, and cell can take more liposoluble medicine to raise its utilization rate, ③ can encapsulate liposoluble medicine and water-soluble medicine, ④ wraps medicine physically without destroying medicine components, ⑥ wraps medicine to reduce its toxicity to specific parts, ⑦ reduces dosage to make medicine have slow release and controlled release functions, ⑧ can be administered by multiple routes.
From the traditional classical thin film dispersion method of discovering liposome to various new methods, a plurality of preparation methods are derived by combining modern science and technology. Several common methods are as follows:
1) film dispersion method: a process for preparing liposome includes such steps as dissolving lipoid material (such as phosphatide, cholesterol and liposoluble medicine) in organic solvent (chloroform, alcohol, etc), oscillating for dissolving, rotary evaporating for removing organic solvent, vacuum evaporating for removing organic solvent, low-temp drying for volatilizing organic solvent, adding hydrated medium (such as phosphate buffer, physiological saline, glucose injection, etc), oscillating for washing membrane until it drops off, and preparing milky white suspension. The method mainly obtains the unilamellar liposomes, the particle size of the liposomes is larger, and the particle size can be reduced and distributed uniformly by utilizing technical means such as probe ultrasound, a high-pressure homogenizer, high-pressure microjet and the like; finally, the liposome can be extruded through a filter membrane, so that the particle size of the obtained liposome is more uniform. The method is simple and easy to operate, but has the defects of organic solvent residue and large particle size difference of the liposome. Wumin and the like adopt a membrane dispersion-ultrasound-membrane filtration method to prepare the magnolia flower volatile oil nano-liposome, and the obtained liposome has good dispersibility and the particle size is mainly distributed in (35 +/-5) nm.
2) Ethanol injection method: dissolving lipid material such as phospholipid and cholesterol in ethanol, and mixing
And (3) quickly injecting ethanol into the aqueous phase medium by using an injector, and performing rotary evaporation under reduced pressure until the ethanol is completely volatilized to obtain the liposome. The obtained liposome is mostly multi-compartment liposome, and the obtained liposome can be sized uniformly as the above method. The method is simple to operate and does not use toxic reagents. The disadvantages are that the reduced pressure distillation of ethanol takes a long time and has a small amount of ethanol residue, and the precondition is that the included medicine must have good solubility in ethanol. Wu Yani et al adopts an ethanol injection method to prepare the papain nano-liposome, and the prepared papain nano-liposome has the average particle size of less than 100nm, high particle circularity, clear outline, regular shape and mutual dispersion.
3) The multiple emulsion method comprises the following steps: the first emulsification prepares W/O type emulsion, the second emulsification prepares W/O/W emulsion. Dissolving a lipid material in an organic solvent, adding a proper amount of a hydration medium, and oscillating or ultrasonically emulsifying to form a W/O type emulsion; adding a large amount of hydration medium (PBS or normal saline, etc.) into the emulsion prepared for the first time, and performing rotary evaporation and reduced pressure evaporation to obtain the liposome. This method is complicated and has many uncertainty factors.
4) Reverse phase evaporation method: dissolving lecithin, cholesterol, fat-soluble medicine and surfactant in an organic solvent according to a proper proportion, then adding a hydration medium, stirring or carrying out ultrasonic treatment to obtain a stable W/O type emulsion, carrying out reduced pressure evaporation or nitrogen filling to remove the organic solvent to obtain a water-soluble suspension, then adding a proper amount of hydration medium, and stirring or carrying out ultrasonic treatment to obtain the liposome. The experiments also show that the bioavailability of the oleanolic acid is obviously improved, and the retention time of the oleanolic acid in vivo is prolonged.
5) And others: freeze thawing method, and extending film dispersing method, to obtain liposome, freezing in refrigerator, taking out, dissolving, and repeating for three times.
When the particle size and uniformity of the liposome are treated, the particle size needs to be reduced by means of ultrasound, but if the power of ultrasound is too low, the whole particle effect cannot be achieved, and if the power of ultrasound is too high, the ultrasound is too long, or the time is too long, or the temperature is too high, the liposome can be damaged and leaked, so that an appropriate ultrasound power, an appropriate ultrasound time and an appropriate temperature need to be found in the preparation process.
The application of the liposome in the pharmaceutical industry is rapidly researched and developed in recent years, and the liposome serving as a drug carrier can be applied to the aspects of tumor resistance, cancer resistance, parasite resistance, immune activators, genetic engineering, gene therapy and the like. With the development and improvement of the preparation technology, the leading direction of liposome technology research is as follows: as artificial blood substitute, as ophthalmic administration carrier, as intrapulmonary controlled release drug delivery system for percutaneous absorption therapy system, for cardiovascular disease diagnosis and treatment.
Detailed Description
The present invention is further illustrated by the following examples, which are not intended to be limiting in any way, and any modifications or alterations based on the teachings of the present invention are intended to fall within the scope of the present invention.
The artemether nanoliposome is prepared from the following raw materials in parts by weight: 1 part of artemether, 2 parts of cholesterol and 8-10 parts of egg yolk lecithin.
The preparation method of the artemether nanoliposome is prepared by any one of the following methods:
(1) thin film dispersion-ultrasonic-membrane filtration method: dissolving artemether, cholesterol and egg yolk lecithin in an organic solvent, performing ultrasonic oscillation to dissolve the artemether, performing rotary evaporation and reduced pressure evaporation to remove the organic solvent after complete dissolution to form a film, adding water for injection to hydrate the film until the film falls off and dissolves, performing ultrasonic treatment for 180s (ultrasonic 1s, intermittent 1 s), filtering the film through a 0.45 mu m filter membrane, and filtering the film through a 0.22 mu m filter membrane to obtain the target artemether nanoliposome;
(2) ethanol injection-sonication: dissolving artemether, cholesterol and egg yolk lecithin in an organic solvent, uniformly dissolving by ultrasonic oscillation, obtaining a solution a after complete dissolution, dropwise adding the solution a into a 5% glucose injection containing tween-80, stirring continuously, oscillating and hydrating at 40 ℃ to obtain a suspension b, evaporating the organic solvent in the suspension b by rotary evaporation and reduced pressure to obtain a crude liposome, carrying out ultrasonic treatment for 120s (ultrasonic 1s, intermittent 1 s), and filtering with a 0.22 mu m filter membrane for 2-4 times to obtain the target artemether nanoliposome.
(1) The organic solvent is absolute ethyl alcohol.
(1) The ultrasonic power of the ultrasonic treatment in (1) is 40%, and the ultrasonic temperature is 40 ℃.
(1) The adding amount of the water for injection is 10-20 ml.
(2) The organic solvent is absolute ethyl alcohol.
(2) The ultrasonic power of the ultrasonic treatment in (1) is 40%, and the ultrasonic temperature is 40 ℃.
(2) The adding amount of the tween-80-containing 5% glucose injection is 10-20 ml.
The adding amount of the Tween in the 5% glucose injection containing the Tween-80 is 5% -20% of the amount of the phospholipid.
The application of the artemether nano-liposome is the application of the artemether nano-liposome in preparing artemether targeted preparation medicaments.
The invention is further illustrated by the following specific examples:
example 1
The artemether nano liposome is mainly prepared from the following raw materials, by weight, 10mg of artemether, 20mg of cholesterol and 80mg of egg yolk lecithin. The membrane is prepared by adopting a membrane dispersion-ultrasound-membrane filtration method, and the preparation method comprises the following specific operations: putting artemether, cholesterol and yolk lecithin into an eggplant-shaped bottle, adding 5-10 ml of absolute ethyl alcohol, carrying out ultrasonic oscillation to dissolve the artemether uniformly, after the dissolution is complete, putting the eggplant-shaped bottle into a constant-temperature water bath at 40 ℃, carrying out reduced pressure rotary evaporation to remove the absolute ethyl alcohol, forming a layer of lipid film on the inner wall of the bottle, slowly adding 10ml of water for injection to hydrate the lipid film until the lipid film falls off and dissolves, carrying out ultrasonic treatment (ultrasonic power is 40%, ultrasonic temperature is 40 ℃) for 180s, carrying out ultrasonic treatment for 1s, carrying out intermittent treatment for 1s, filtering through a 0.45 mu m filter membrane, and then filtering through a 0.22 mu m filter membrane to obtain the target artemether nanoliposome.
Through detection, the artemether nanoliposome prepared in example 1 is milky white and blue opalescent suspension, the average particle size is 161.65nm, the encapsulation efficiency is 61.33%, and the dispersion index (PID) is 0.4463.
Example 2
The artemether nano liposome is mainly prepared from the following raw materials, by weight, 10mg of artemether, 20mg of cholesterol and 120mg of egg yolk lecithin. The membrane is prepared by adopting a membrane dispersion-ultrasound-membrane filtration method, and the preparation method comprises the following specific operations: putting artemether, cholesterol and yolk lecithin into an eggplant-shaped bottle, adding 5-10 ml of absolute ethyl alcohol, carrying out ultrasonic oscillation to dissolve the artemether uniformly, after the dissolution is complete, putting the eggplant-shaped bottle into a constant-temperature water bath at 40 ℃, carrying out reduced pressure rotary evaporation to remove the absolute ethyl alcohol, forming a layer of lipid film on the inner wall of the bottle, slowly adding 10ml of water for injection to hydrate the lipid film until the lipid film falls off and dissolves, carrying out ultrasonic treatment (ultrasonic power is 40%, ultrasonic temperature is 40 ℃) for 180s, carrying out ultrasonic treatment for 1s, carrying out intermittent treatment for 1s, filtering through a 0.45 mu m filter membrane, and then filtering through a 0.22 mu m filter membrane to obtain the target artemether nanoliposome.
Through detection, the artemether nanoliposome prepared in example 2 is milky white and blue opalescent suspension, the average particle size is 185.1nm, the encapsulation efficiency is 62.90%, and the dispersion index (PID) is 0.8561.
Example 3
The artemether nano liposome is mainly prepared from the following raw materials, by weight, 10mg of artemether, 20mg of cholesterol and 80mg of egg yolk lecithin. The membrane is prepared by adopting a membrane dispersion-ultrasound-membrane filtration method, and the preparation method comprises the following specific operations: putting artemether, cholesterol and yolk lecithin into an eggplant-shaped bottle, adding 5-10 ml of absolute ethyl alcohol, carrying out ultrasonic oscillation to dissolve the artemether uniformly, after the dissolution is complete, putting the eggplant-shaped bottle into a constant-temperature water bath at 40 ℃, carrying out reduced pressure rotary evaporation to remove the absolute ethyl alcohol, forming a layer of lipid film on the inner wall of the bottle, slowly adding 5ml of water for injection to hydrate the lipid film until the lipid film falls off and dissolves, carrying out ultrasonic treatment (ultrasonic power is 40%, ultrasonic temperature is 40 ℃) for 180s, carrying out ultrasonic treatment for 1s, carrying out intermittent treatment for 1s, filtering through a 0.45 mu m filter membrane, and then filtering through a 0.22 mu m filter membrane to obtain the target artemether nanoliposome.
Through detection, the artemether nanoliposome prepared in example 3 is milky white suspension with blue opalescence, the average particle size is 184.4nm, the encapsulation efficiency is 60.74%, and the dispersion index (PID) is 0.7349.
Example 4
The artemether nano liposome is mainly prepared from the following raw materials, by weight, 10mg of artemether, 20mg of cholesterol and 100mg of egg yolk lecithin. The preparation method is characterized by comprising the following specific operations: putting artemether, cholesterol and yolk lecithin into an eggplant-shaped bottle, adding 5-10 ml of absolute ethyl alcohol, carrying out ultrasonic oscillation to dissolve uniformly to obtain a solution a after complete dissolution, dropwise adding the solution a into 10ml of 5% glucose injection containing 10mg of tween-80 (10% of phospholipid), stirring continuously, carrying out oscillation hydration for 60min at 40 ℃ to obtain a suspension b, carrying out reduced pressure rotary evaporation for 45-120 min in a constant-temperature water bath at 40 ℃ to volatilize the absolute ethyl alcohol to obtain a crude liposome, carrying out ultrasonic treatment (the ultrasonic power is 40%, the ultrasonic temperature is 40 ℃) for 120s, carrying out ultrasonic treatment for 1s, carrying out intermittent 1s, and filtering a filter membrane for 4 times at 0.22 mu m to obtain the target artemether nanoliposome.
Through detection, the artemether nanoliposome prepared in example 4 is semitransparent liquid with blue opalescence, the average particle size is 162.73nm, the encapsulation efficiency is 63.86%, and the dispersion index (PID) is 0.2678.
Example 5
The artemether nano liposome is mainly prepared from the following raw materials, by weight, 10mg of artemether, 20mg of cholesterol and 140mg of egg yolk lecithin. The preparation method is characterized by comprising the following specific operations: putting artemether, cholesterol and yolk lecithin into an eggplant-shaped bottle, adding 5-10 ml of absolute ethyl alcohol, performing ultrasonic oscillation to dissolve uniformly to obtain a solution a after complete dissolution, dropwise adding the solution a into 10ml of 5% glucose injection containing 10mg of Tween-80, stirring continuously, performing oscillation hydration for 60min at 40 ℃ to obtain a suspension b, performing reduced pressure rotary evaporation for 45-120 min in a constant-temperature water bath at 40 ℃ to volatilize the absolute ethyl alcohol to obtain a crude liposome, performing ultrasonic treatment (the ultrasonic power is 40%, the ultrasonic temperature is 40 ℃) for 120s, performing ultrasonic treatment for 1s, performing intermittent 1s, and filtering with a 0.22 mu m filter membrane for 4 times to obtain the target artemether nano-liposome.
Through detection, the artemether nanoliposome prepared in example 5 is a semitransparent liquid with blue opalescence, the average particle size is 127.9nm, the entrapment rate is 52.41%, and the dispersion index (PID) is 0.2591.
Example 6 phospholipid concentration differences
The artemether nano liposome is mainly prepared from the following raw materials, by weight, 10mg of artemether, 20mg of cholesterol and 100mg of egg yolk lecithin. The preparation method is characterized by comprising the following specific operations: putting artemether, cholesterol and yolk lecithin into an eggplant-shaped bottle, adding 5-10 ml of absolute ethyl alcohol, performing ultrasonic oscillation to dissolve uniformly to obtain a solution a after complete dissolution, dropwise adding the solution a into 5ml of 5% glucose injection containing 10mg of Tween-80, stirring continuously, performing oscillation hydration for 60min at 40 ℃ to obtain a suspension b, performing reduced pressure rotary evaporation for 45-120 min in a constant-temperature water bath at 40 ℃ to volatilize the absolute ethyl alcohol to obtain a crude liposome, performing ultrasonic treatment (the ultrasonic power is 40%, the ultrasonic temperature is 40 ℃) for 120s, performing ultrasonic treatment for 1s, performing intermittent 1s, and filtering with a 0.22 mu m filter membrane for 4 times to obtain the target artemether nano-liposome.
Through detection, the artemether nanoliposome prepared in example 6 is a transparent liquid with blue opalescence, the average particle size is 214.8nm, the entrapment rate is 72.33%, and the dispersion index (PID) is 0.6423.
Example 7 different% of Tween-80 based on phospholipid
The artemether nano liposome is mainly prepared from the following raw materials, by weight, 10mg of artemether, 20mg of cholesterol and 10mg of egg yolk lecithin. The preparation method is characterized by comprising the following specific operations: putting artemether, cholesterol and yolk lecithin into an eggplant-shaped bottle, adding 5-10 ml of absolute ethyl alcohol, carrying out ultrasonic oscillation to dissolve uniformly to obtain a solution a after complete dissolution, dropwise adding the solution a into 5ml of 5% glucose injection containing 20mg of tween-80 (20% of phospholipid), stirring continuously, carrying out oscillation hydration for 60min at 40 ℃ to obtain a suspension b, carrying out reduced pressure rotary evaporation for 45-120 min in a constant-temperature water bath at 40 ℃ to volatilize the absolute ethyl alcohol to obtain a crude liposome, carrying out ultrasonic treatment (the ultrasonic power is 40%, the ultrasonic temperature is 40 ℃) for 120s, carrying out ultrasonic treatment for 1s, carrying out intermittent 1s, and filtering a filter membrane for 4 times at 0.22 mu m to obtain the target artemether nanoliposome.
Through detection, the artemether nanoliposome prepared in example 6 is a transparent liquid with blue opalescence, the average particle size is 165.9nm, the encapsulation efficiency is 60.48%, and the dispersion index (PID) is 0.4741.
Claims (5)
1. The preparation method of the artemether nanoliposome is characterized in that the artemether nanoliposome is prepared from 1 part by weight of artemether, 2 parts by weight of cholesterol and 8-10 parts by weight of egg yolk lecithin by adopting an ethanol injection-ultrasonic method: dissolving artemether, cholesterol and egg yolk lecithin in absolute ethyl alcohol, performing ultrasonic oscillation to dissolve the artemether, dissolving the artemether, the cholesterol and the egg yolk lecithin uniformly to obtain a solution a completely, dripping the solution a into a 5% glucose injection containing tween-80, stirring continuously, performing oscillation hydration at 40 ℃ to obtain a suspension b, performing rotary evaporation and reduced pressure evaporation to remove the absolute ethyl alcohol in the suspension b to obtain a crude liposome, performing ultrasonic treatment for 120s in an ultrasonic mode of 1s and 1s at intervals, wherein the ultrasonic power is 40%, the ultrasonic temperature is 40 ℃, and filtering with a 0.22 mu m filter membrane for 2-4 times to obtain the target artemether nanoliposome.
2. The method for preparing artemether nanoliposomes according to claim 1, wherein the amount of the 5% glucose injection containing tween-80 is 10-20 ml.
3. The method for preparing artemether nanoliposomes according to claim 1, wherein the amount of tween added to the 5% glucose injection containing tween-80 is 5% to 20% of the weight of egg yolk lecithin.
4. An artemether nanoliposome prepared by the preparation method of claim 1.
5. An application of the artemether nanoliposome of claim 4 in preparing artemether targeted preparation medicine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510533475.1A CN106474064B (en) | 2015-08-27 | 2015-08-27 | Artemether nanoliposome and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510533475.1A CN106474064B (en) | 2015-08-27 | 2015-08-27 | Artemether nanoliposome and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106474064A CN106474064A (en) | 2017-03-08 |
| CN106474064B true CN106474064B (en) | 2020-07-28 |
Family
ID=58234587
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510533475.1A Active CN106474064B (en) | 2015-08-27 | 2015-08-27 | Artemether nanoliposome and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106474064B (en) |
Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107669637B (en) * | 2017-09-30 | 2021-11-02 | 山东大学 | Artemether liposome for injection and preparation method and application thereof |
| CN109893664B (en) * | 2017-12-11 | 2022-03-11 | 四川大学 | Cationic lipid nanoparticle/DNA compound and preparation method thereof |
| WO2020109418A1 (en) * | 2018-11-27 | 2020-06-04 | Avignon Universite | Method for extracting substances of interest |
| CN113181350B (en) * | 2021-04-27 | 2023-04-25 | 浙江大学 | Peptide vaccine based on activated fibroblast growth protein (FAP) and preparation method thereof |
| CN113521006B (en) * | 2021-07-14 | 2023-04-28 | 广州中医药大学(广州中医药研究院) | Artemether liposome, erythrocyte membrane-encapsulated liposome, targeting peptide-modified biomimetic liposome, preparation method thereof and application of targeting peptide-modified biomimetic liposome in treating malaria |
| CN113975234B (en) * | 2021-09-26 | 2023-01-31 | 成都中医药大学 | Hydroxyl-alpha-sanshool nano liposome and preparation method thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101244027A (en) * | 2008-03-20 | 2008-08-20 | 昆明制药集团股份有限公司 | Artemether percutaneous drug administration preparation for treating dermatosis |
| CN103417530A (en) * | 2013-09-03 | 2013-12-04 | 福州大学 | Method for applying artemisinin derivative and lipidosome of artemisinin derivative to preparation of sound sensitive agent |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2002069949A2 (en) * | 2001-03-06 | 2002-09-12 | Prendergast Patrick T | Combination therapy for reduction of toxycity of chemotherapeutic agents |
-
2015
- 2015-08-27 CN CN201510533475.1A patent/CN106474064B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101244027A (en) * | 2008-03-20 | 2008-08-20 | 昆明制药集团股份有限公司 | Artemether percutaneous drug administration preparation for treating dermatosis |
| CN103417530A (en) * | 2013-09-03 | 2013-12-04 | 福州大学 | Method for applying artemisinin derivative and lipidosome of artemisinin derivative to preparation of sound sensitive agent |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106474064A (en) | 2017-03-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Majumder et al. | Nanocarrier-based systems for targeted and site specific therapeutic delivery | |
| CN106474064B (en) | Artemether nanoliposome and preparation method and application thereof | |
| Dal Magro et al. | ApoE-modified solid lipid nanoparticles: A feasible strategy to cross the blood-brain barrier | |
| Gulati et al. | Parenteral drug delivery: a review | |
| Jing et al. | A novel polyethylene glycol mediated lipid nanoemulsion as drug delivery carrier for paclitaxel | |
| US20080279764A1 (en) | Method and composition for treating cancer | |
| Kushwaha et al. | Novel drug delivery system for anticancer drug: a review | |
| CN102188377B (en) | Method for preparing medicine encapsulating liposome | |
| WO2020203961A1 (en) | Lipid membrane structure and manufacturing method therefor | |
| CN101953792B (en) | Irinotecan nano circulating liposome and preparation method thereof | |
| Bandawane et al. | A review on novel drug delivery system: a recent trend | |
| Holsæter et al. | How docetaxel entrapment, vesicle size, zeta potential and stability change with liposome composition–A formulation screening study | |
| CN111920768A (en) | Entrapped molecular targeted drug liposome and application thereof in preparation of tumor treatment drug | |
| Kumar et al. | Novel drug delivery system | |
| Bangale et al. | Stealth liposomes: a novel approach of targeted drug delivery in cancer therapy | |
| Cai et al. | Lipid nanoparticle steric stabilization roadmap | |
| Upadhyay et al. | Vesicular approach review on nanocarriers bearing curcumin and applications | |
| Pagar et al. | A review on novel drug delivery system: a recent trend | |
| Tian et al. | Preparation and evaluation of artemether liposomes for enhanced anti-tumor therapy | |
| Patel et al. | Niosome: a vesicular drug delivery tool | |
| Chandra et al. | An overview: The novel carrier for vesicular drug delivery system | |
| CN114712520A (en) | Nanocrystalline drug stabilizing system, preparation method, pharmaceutical composition and application | |
| Liao et al. | Preparation of galactosyl nanoparticles and their targeting efficiency to hepatocellular carcinoma | |
| Jing-Jing et al. | Fabrication of a folic acid-modified arsenic trioxide prodrug liposome and assessment of its anti-hepatocellular carcinoma activity | |
| Binaymotlagh et al. | Liposome–Hydrogel Composites for Controlled Drug Delivery Applications |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20210802 Address after: No. 166, Keyi Road, national high tech Development Zone, Wuhua District, Kunming, Yunnan 650106 Patentee after: Kunming Pharmaceutical Corp. Patentee after: Shanghai kunheng Medical Technology Co.,Ltd. Address before: 650106 No. 166, medical Road, hi tech Industrial Development Zone, Yunnan, Kunming Patentee before: Kunming Pharmaceutical Corp. |
|
| TR01 | Transfer of patent right |