CN113786386B - ROS (reactive oxygen species) -responsive mitochondrial targeting quercetin liposome as well as preparation method and application thereof - Google Patents
ROS (reactive oxygen species) -responsive mitochondrial targeting quercetin liposome as well as preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a multifunctional quercetin liposome based on ROS response and mitochondrion targeting, and a preparation method and application thereof. According to the invention, ROS response element Di-S-GPC and traditional soybean lecithin are used as liposome shell materials according to a proper proportion to wrap the hydrophobic drug quercetin, and the hydrophobic drug quercetin is prepared into a hydrophilic preparation, so that the problem of difficulty in drug formation of the quercetin is solved, the slow release effect is achieved, the cell membrane penetration capacity is enhanced, and simultaneously the peak valley phenomenon caused by direct drug delivery and the toxic and side effects of the quercetin are reduced.
Description
Technical Field
The invention relates to a liposome, a preparation method and application thereof, in particular to a multifunctional quercetin liposome based on ROS response and mitochondrial targeting, a preparation method and application thereof, and belongs to the technical field of liposomes.
Background
Retinal ischemia reperfusion injury is one of the leading causes of blindness and is mainly characterized by apoptosis and loss of function of the optic nerve cells (RGCs) in the retina. The main measures for preventing or treating retinal ischemia-reperfusion injury include:
1. eliminating ROS, inhibiting apoptosis of optic nerve cells after ischemia reperfusion;
2. inhibiting the occurrence of retinal inflammation;
3. promoting the repair of the function of the optic nerve cells.
Although the clearance of ROS and the inhibition of inflammatory responses are considered to be the major approaches to the treatment of retinal ischemia reperfusion injury, there is currently a lack of effective drugs for the treatment of retinal ischemia reperfusion injury.
Quercetin is a natural antioxidant anti-inflammatory drug and has functions of ROS removal and inflammation inhibition, however, quercetin as a fat-soluble compound has extremely poor water solubility which severely limits the clinical application of quercetin, and the currently reported quercetin liposome has the problem of lack of targeting property, which is not favorable for the full play of the anti-inflammatory and anti-oxidation capacity of quercetin.
Disclosure of Invention
The invention aims to provide a multifunctional quercetin liposome based on ROS response and mitochondrion targeting, and a preparation method and application thereof.
In order to achieve the above object, the present invention adopts the following technical solutions:
a preparation method of ROS-responsive mitochondrial targeting quercetin liposome is characterized by comprising the following steps:
step 1, synthesizing ROS response element
The ROS response element is Di-S-GPC, and the step of synthesizing the Di-S-GPC is as follows:
(1) And (2) mixing carbonyldiimidazole and C18-S-COOH according to a mass ratio of 3:4, adding the mixture into dichloromethane to obtain a first-phase solution;
(2) Adding 1,8-diazabicyclo [5.4.0] undec-7-ene and glycerophosphatidylcholine into dimethyl sulfoxide according to the mass ratio of 1:1 to obtain a second-phase solution;
(3) Stirring the first phase solution and the second phase solution at 30 ℃ for 1h respectively, transferring the first phase solution into the second phase solution, and further reacting at 30 ℃ for 24h to obtain a crude product;
(4) Purifying the crude product by gradient elution silica gel column chromatography to obtain Di-S-GPC lipid;
The method for preparing the ROS-responsive quercetin liposome comprises the following steps of:
(1) Mixing soybean lecithin, di-S-GPC, cholesterol, distearoyl phosphatidyl ethanolamine-polyethylene glycol-2000 and quercetin according to the mass ratio of 17-18: 5:5 to 6:1 to 2:1, mixing to obtain a mixture with a medicine-fat ratio of 1;
(2) Placing the yellow clear solution prepared in the previous step in a rotary evaporator at 37 ℃ for rotary evaporation until a yellow film is formed, then adding ultrapure water, then placing the yellow clear solution in a rotary evaporator at 50 ℃ for rotary evaporation hydration, transferring the obtained solution into a clean penicillin bottle after hydration, and crushing the solution by using a cell crusher to crush larger particles into small particles to finally obtain yellow transparent liquid;
(3) Subpackaging the yellow transparent liquid prepared in the previous step into an EP tube, placing the EP tube into a centrifugal machine for centrifugation to obtain the quercetin nanoliposome with higher purity, and transferring the supernatant solution in the EP tube into a clean penicillin bottle;
(4) Placing the penicillin bottle filled with the supernatant solution into a cell disruptor, and ultrasonically controlling the particle size of the liposome by using the cell disruptor again to finally obtain a yellow clarified liposome solution, namely an ROS-responsive quercetin nanoliposome solution;
The steps of adding the mitochondrion targeting molecule into the ROS-responsive quercetin liposome are as follows:
polyethylene glycol modified triphenyl phosphonic acid-2000 and ROS response type quercetin liposome prepared in the last step are added according to the weight ratio of 2mg: placing 3ml of the mixture into a round-bottom flask, heating the mixture in a water bath at 50 ℃ and continuously stirring the mixture by a magnetic rotor for 2 hours to obtain the ROS-responsive type mitochondria targeted quercetin liposome containing triphenyl phosphate molecules.
The preparation method of the ROS-responsive mitochondrial targeting quercetin liposome is characterized in that in the step 1, the synthesis method of the C18-S-COOH comprises the following steps:
adding mercaptoacetic acid and bromooctadecane into a 25% potassium hydroxide methanol solution, vigorously stirring at room temperature for 48h, then acidifying with hydrochloric acid to pH =1, and then sequentially performing ethyl acetate extraction, anhydrous sodium sulfate drying and silica gel column chromatography purification to obtain C18-S-COOH.
The preparation method of the ROS-responsive mitochondrial targeting quercetin liposome is characterized in that the molar ratio of the thioglycolic acid to the bromooctadecane is 1:1.
The preparation method of the ROS-responsive mitochondrial targeting quercetin liposome is characterized in that when silica gel column chromatography purification is carried out, hexane/ethyl acetate 3.
The preparation method of the ROS-responsive mitochondrial targeting quercetin liposome is characterized in that in the step 1, when a crude product is purified by a gradient elution silica gel column chromatography, the eluent A adopts dichloromethane: methanol 5; the eluent B used was dichloromethane: methanol: water 65.
The preparation method of the ROS response type mitochondrial targeting quercetin liposome is characterized in that in the step 2, chloroform and methanol are mixed according to the volume ratio of 3:1 in a chloroform/methanol mixed solvent.
The ROS response type mitochondrial targeting quercetin liposome is characterized by being prepared by the method, and the ROS response type mitochondrial targeting quercetin liposome can be applied to vitreous injection treatment of retinal ischemia reperfusion injury.
The invention has the advantages that:
(1) According to the invention, a ROS response element Di-S-GPC and traditional soybean lecithin are used as liposome shell materials according to a proper proportion to wrap a hydrophobic drug quercetin, and the hydrophobic drug quercetin is prepared into a hydrophilic preparation, so that the drug forming property of the quercetin is improved, and the problem of difficulty in forming the drug of the quercetin is solved;
(2) The invention wraps the quercetin by the liposome, thereby playing a role of slow release, prolonging the metabolism time of the quercetin in vivo, enhancing the penetrating capability of cell membranes, and simultaneously reducing the peak-valley phenomenon caused by direct administration and the toxic and side effects of the quercetin;
(3) According to the invention, by introducing ROS response groups and mitochondrion targeting molecules TPP, quercetin can be selectively released in a targeted manner at an ROS enrichment position, so that the anti-inflammatory capacity and the antioxidant capacity of quercetin are enhanced;
(4) The quercetin liposome prepared by the invention can effectively inhibit the activation and proliferation of retinal inflammatory cells, has good treatment effect on retinal ischemia reperfusion injury, and can be applied to the treatment of retinal ischemia reperfusion injury by vitreous injection.
Drawings
FIG. 1 is a synthetic scheme for ROS response element Di-S-GPC;
FIG. 2 is of ROS response element Di-S-GPC 1 H-NMR chart;
FIG. 3 is a MS plot of ROS response elements Di-S-GPC;
fig. 4 (a), fig. 4 (b), and fig. 4 (c) are electron micrograph, particle size distribution plot, and Zeta potential plot, respectively, of ROS responsive mitochondrial targeting quercetin liposomes;
fig. 5 is a graph of the in vitro release of ROS responsive mitochondrial targeting quercetin liposomes;
FIG. 6 is a photograph of uptake of retinal progenitor cells to ROS responsive mitochondrial targeting of quercetin liposomes;
fig. 7 is a mitochondrial targeting profile after ROS-responsive mitochondrial targeting of quercetin liposomes into retinal progenitor cells;
FIG. 8 is a primary macrophage inflammatory factor secretion assay;
FIG. 9 is a graph of retinal progenitor ATP assay;
FIG. 10 is an immunofluorescence assay of rat retinal microglia;
FIG. 11 is a graph of HE staining of rat retinas;
FIG. 12 is an immunofluorescence assay of rat retinal β -III-Tubulin protein.
Detailed Description
The invention is described in detail below with reference to the figures and the embodiments.
First part, preparation of ROS-responsive type mitochondrial targeting quercetin liposome
The raw materials comprise: quercetin (bulk drug), soybean lecithin (liposome shell material), cholesterol (membrane fluidity regulator), di-S-GPC (ROS response element), distearoylphosphatidylethanolamine-polyethylene glycol-2000 (DSPE-PEG-2000, long-circulating protective agent), polyethylene glycol modified triphenyl phosphonic acid-2000 (TPP-PEG-2000, mitochondrion targeting molecule)
The preparation method of the ROS response type mitochondrial targeting quercetin liposome specifically comprises the following steps:
step 1, synthesizing ROS response element Di-S-GPC
Di-S-GPC has the following structural formula:
referring to FIG. 1, the steps for synthesizing Di-S-GPC are specifically as follows:
(1) Mercaptoacetic acid (1.12g, 12.02mmol) and bromooctadecane (4 g, 12.02mmol) were added to a 25% strength by mass potassium hydroxide methanol solution (10 ml), stirred vigorously at room temperature for 48h, then acidified to pH =1 with aqueous HCl (0.1M), followed by ethyl acetate extraction, anhydrous sodium sulfate drying and purification by silica gel column chromatography (eluent hexane/ethyl acetate 3:1, volume ratio) in this order to give C18-S-COOH (1.72 g, yield 41%) as a white solid.
(2) Carbonyldiimidazole (0.71g, 4.36mmol) and C18-S-COOH (1g, 2.91mmol) prepared in the previous step were added to dichloromethane (15 ml) to obtain a first phase solution; 1,8-diazabicyclo [5.4.0] undec-7-ene (0.29g, 1.94mmol) and glycerophosphatidylcholine (0.3g, 1.16mmol) were added to dimethyl sulfoxide (15 ml) to give a second phase solution; stirring the first phase solution and the second phase solution at 30 ℃ for 1h respectively, transferring the first phase solution into the second phase solution, and further reacting at 30 ℃ for 24h to obtain a crude product; without any work-up, the crude product was purified by gradient elution silica gel column chromatography (eluent a: dichloromethane: methanol 5:1, vol.; eluent B: dichloromethane: methanol: water 65, vol..
Use of 1 The structure of the obtained light yellow solid is characterized by H-NMR and MS means, and the characterization results are shown in figures 2 and 3, and figures 2 and 3 show that: SDi-S-GPC was synthesized successfully.
(1) Soybean lecithin 53mg, di-S-GPC 15mg, cholesterol 17mg, distearoylphosphatidylethanolamine-polyethylene glycol-2000 (DSPE-PEG-2000) 4.8mg, and quercetin 3mg were precisely weighed to obtain a mixture having a drug-to-lipid ratio of 1.
(2) Placing the yellow clear solution prepared in the previous step on a rotary evaporator at 37 ℃ and 90r/min for rotary evaporation until a yellow film is formed, then adding 8ml of ultrapure water, then placing the yellow clear solution on a rotary evaporator at 50 ℃ and 100r/min for rotary evaporation and hydration for 50min, wherein negative pressure treatment is not needed in the process, transferring the obtained solution into a clean penicillin bottle after hydration, crushing the solution by using a cell crusher to crush larger particles into small particles, pausing for 5s every 5s of ultrasonic treatment for 2min, and finally obtaining yellow transparent liquid.
(3) Subpackaging the yellow transparent liquid prepared in the previous step into 1.5ml EP tubes, placing in a centrifuge, centrifuging at a speed of 5000r/min for 10min, sufficiently separating quercetin which is not wrapped into the liposome from the quercetin nanoliposome to obtain the quercetin nanoliposome with higher purity, and transferring the supernatant solution in the EP tubes into a clean penicillin bottle.
(4) And (3) placing the penicillin bottle filled with the supernatant solution into a cell disruptor, ultrasonically controlling the particle size of the liposome by using the cell disruptor again, pausing for 5s every 5s of ultrasonic treatment for 10min, and controlling the particle size of the liposome to be 140-180 nm to finally obtain a yellow clarified liposome solution, namely the ROS-responsive quercetin nano-liposome solution.
Storing the prepared ROS response type quercetin nanoliposome at 4 ℃ for later use.
Measuring the encapsulation rate of the quercetin liposome: the quercetin liposomes were dissolved in methanol, and the concentration of the quercetin liposomes was measured by an ultraviolet-visible spectrophotometer at a wavelength of 372nm using a Perkin Elmer Lambda 6 ultraviolet-visible spectrometer manufactured by Perkin Elmer, boston, mass. The encapsulation efficiency of the quercetin liposome is calculated to be 85.3%.
The step of adding the mitochondrial targeting molecule TPP to the ROS-responsive quercetin liposome is as follows:
accurately weighing 2mg of polyethylene glycol modified triphenyl phosphonic acid-2000 (TPP-PEG-2000), placing the weighed 2mg into a 50ml round-bottom flask, adding 3ml of ROS response type quercetin liposome prepared in the previous step, placing the round-bottom flask into a 50 ℃ water bath kettle for heating, and continuously stirring for 2 hours by using a magnetic rotor to obtain the ROS response type mitochondrial targeting quercetin liposome containing triphenyl phosphate (TPP) molecules.
Second, characterization of ROS-responsive mitochondrial-targeted quercetin liposomes
1. Particle diameter, zeta potential and morphology
And characterizing the particle size and the Zeta potential of the ROS response type mitochondrial targeting quercetin liposome prepared by the first part by a particle size analyzer.
And characterizing the morphology of the ROS-responsive type mitochondrial targeting quercetin liposome prepared by the first part by a Transmission Electron Microscope (TEM).
The characterization results are shown in FIG. 4 (a), FIG. 4 (b) and FIG. 4 (c). As can be seen from fig. 4 (a), 4 (b) and 4 (c): the ROS response type mitochondrial targeting quercetin liposome is regular spherical, has uniform size, particle size of about 148nm, zeta potential of-27.07 mV, good dispersibility and high stability.
2. In vitro release
Placing ROS response type mitochondria targeting quercetin liposome prepared from the first part into a dialysis bag with molecular weight of 500, and adding 40ml of Tween-80 with mass concentration of 1% and H with different concentrations 2 O 2 The PBS is used as a release medium, and the in vitro release condition of the drug quercetin is detected in a shaking table with 37 ℃ and 120 rpm. The release medium of each sample was replaced at fixed time points with an equal amount of fresh release medium, 1ml each time. And (3) passing the obtained sample through a 0.22-micron filter membrane, detecting the content of the medicine quercetin by using HPLC, and finally calculating the accumulated release amount according to a standard curve of the medicine quercetin.
The in vitro release results are shown in figure 5. As can be seen from fig. 5: after the quercetin is encapsulated in the liposome, the release amount is time-dependent, and H in the medium is released 2 O 2 The higher the content, the faster the release rate of quercetin, indicating that the liposome has the capability of prolonging the drug release time and the capability of ROS response.
Third, cytological study of ROS-responsive mitochondria-targeted quercetin liposomes
1. Cell uptake assay
Taking 3ml of ROS response type mitochondrial targeting quercetin liposome prepared in the first part, placing the ROS response type mitochondrial targeting quercetin liposome in a 50ml round-bottom flask, adding FITC-PEG-2000 5mg into the round-bottom flask, and then heating in a water bath at 50 ℃ and continuously stirring for 2h to obtain the FITC fluorescence labeled liposome.
Inoculating retinal progenitor cells (R28 cells) into a 6cm cell culture dish, adding the FITC fluorescent labeled liposome after overnight, washing out a culture solution containing the liposome after 24h, labeling cell nuclei through cell fixation, permeation and DAPI, labeling mitochondria by a mitochondrial fluorescent probe Mitotrack-red, and observing the uptake condition of the liposome and the targeting condition of the mitochondria by the cells by using a laser confocal microscope.
The observation results are shown in FIGS. 6 and 7. As can be seen from fig. 6 and 7: the ROS response type mitochondrial targeting quercetin liposome prepared in the first part is successfully taken up by R28 cells, and the distribution condition in the cells is matched with that of the mitochondria, which shows that the ROS response type mitochondrial targeting quercetin liposome has good biocompatibility and mitochondrial targeting property.
2. Study of anti-inflammatory Activity
Inoculating the extracted primary abdominal macrophages to a 24-hole plate, respectively adding LPS stimulating liquid containing ROS response type mitochondria targeting quercetin liposome prepared by the first part with different concentrations after overnight, and detecting the content of IL-1 beta in the supernatant by using an ELISA kit after 24 h.
The results are shown in FIG. 8. As can be seen from fig. 8: the ROS response type mitochondrial targeting quercetin liposome obviously inhibits the generation of inflammatory factors, and the inhibition effect has concentration dependence, so that the ROS response type mitochondrial targeting quercetin liposome prepared by the first part of the invention has good anti-inflammatory effect.
3. Study of cytoprotective ability
R28 cells are inoculated in a 96-well plate, the sugar-free medium containing ROS response type mitochondria targeting quercetin liposome prepared by the first part of the invention with different concentrations is replaced after overnight and is placed in a hypoxic chamber to simulate an ischemic environment, and the cells are taken out after 24h to detect the ATP content in the cells.
The results are shown in FIG. 9. As can be seen from fig. 9: the ROS response type mitochondrial targeting quercetin liposome obviously inhibits the phenomenon that the content of ATP in cells is reduced after hypoxia, and the effect has concentration dependence, so that the ROS response type mitochondrial targeting quercetin liposome prepared by the invention shows good protective capability of retinal progenitor cells in vitro.
Fourth, animal level research of ROS response type mitochondria targeting quercetin liposome
1. Rat retinal inflammation inhibition experiment
Constructing a rat retinal ischemia reperfusion injury model by using an anterior chamber pressurization method, injecting a first part of prepared ROS response type mitochondria targeting quercetin liposome into the vitreous body of a rat, killing the rat after 7d, taking out eyeballs, carrying out frozen sectioning after dehydrating, fixing and embedding the eyeballs, carrying out fluorescent staining on microglia specific protein GFAP and cell nucleuses of the eyeballs, and observing the fluorescent intensity by using a laser confocal microscope.
The observation results are shown in FIG. 10. As can be seen from fig. 10: compared with a control group, after the ROS response type mitochondria targeting quercetin liposome is injected into a vitreous body, the content of the GFAP protein of the microglia is obviously reduced, which shows that the activation proliferation of the microglia is inhibited, and proves that the ROS response type mitochondria targeting quercetin liposome has good retina inflammation inhibition capability in the body.
2. Retinal HE staining
The rat eyeball sections obtained in the previous rat retinal inflammation inhibition experiment were stained with hematoxylin-eosin, and then placed under a microscope to observe the retinal structural morphology.
The observation results are shown in FIG. 11. As can be seen from fig. 11: compared with a control group, after the ROS response type mitochondria targeting quercetin liposome is injected into a vitreous body, the form and the thickness of the retina are well maintained, and the ROS response type mitochondria targeting quercetin liposome successfully inhibits the apoptosis of retina cells after ischemia reperfusion.
3. Optic nerve cell protection experiment
The rat eyeball sections obtained in the previous rat retinal inflammation inhibition experiment were subjected to fluorescence staining of optic nerve cell specific protein β -III-Tubulin and cell nucleus, and the fluorescence intensity was observed using a confocal laser microscope.
The observation results are shown in FIG. 12. As can be seen from fig. 12: compared with a control group, after ROS response type mitochondria targeting quercetin liposome is injected into a vitreous body, the fluorescence intensity of optic nerve cell specific protein beta-III-Tubulin is obviously enhanced, and the ROS response type mitochondria targeting quercetin liposome is proved to play a good role in protecting optic nerve cells after retinal ischemia reperfusion.
It should be noted that the above-mentioned embodiments do not limit the present invention in any way, and all technical solutions obtained by using equivalent alternatives or equivalent variations fall within the protection scope of the present invention.
Claims (4)
1. A preparation method of ROS-responsive mitochondrial targeting quercetin liposome is characterized by comprising the following steps:
step 1, synthesizing ROS response element
The ROS response element is Di-S-GPC, and the step of synthesizing the Di-S-GPC is as follows:
(1) Adding mercaptoacetic acid and bromooctadecane in a molar ratio of 1:1 into a 25% potassium hydroxide methanol solution, vigorously stirring at room temperature for 48h, then acidifying with hydrochloric acid until the pH is =1, then sequentially performing ethyl acetate extraction, anhydrous sodium sulfate drying and silica gel column chromatography purification, wherein an eluent adopts a mixed solution of hexane and ethyl acetate in a volume ratio of 3:1 to obtain C18-S-COOH, and carbonyldiimidazole and C18-S-COOH are mixed according to a mass ratio of 3:4, adding the mixture into dichloromethane to obtain a first-phase solution;
(2) Adding 1,8-diazabicyclo [5.4.0] undec-7-ene and glycerophosphatidylcholine into dimethyl sulfoxide according to the mass ratio of 1:1 to obtain a second-phase solution;
(3) Stirring the first phase solution and the second phase solution at 30 ℃ for 1h respectively, transferring the first phase solution into the second phase solution, and further reacting at 30 ℃ for 24h to obtain a crude product;
(4) Purifying the crude product by gradient elution silica gel column chromatography to obtain Di-S-GPC lipid;
step 2, preparing ROS response type quercetin liposome
The method for preparing the ROS-responsive quercetin liposome comprises the following steps of:
(1) Mixing soybean lecithin, di-S-GPC, cholesterol, distearoyl phosphatidyl ethanolamine-polyethylene glycol-2000 and quercetin according to the mass ratio of 17-18: 5:5 to 6:1 to 2:1 to obtain a mixture with a medicine-fat ratio of 1;
(2) Placing the yellow clear solution prepared in the previous step in a rotary evaporator at 37 ℃ for rotary evaporation until a yellow film is formed, then adding ultrapure water, then placing the yellow clear solution in a rotary evaporator at 50 ℃ for rotary evaporation hydration, transferring the obtained solution into a clean penicillin bottle after hydration, and crushing the solution by using a cell crusher to crush larger particles into small particles to finally obtain yellow transparent liquid;
(3) Subpackaging the yellow transparent liquid prepared in the previous step into an EP tube, placing the EP tube in a centrifuge for centrifugation to obtain quercetin nanoliposome with higher purity, and transferring the supernatant solution in the EP tube into a clean penicillin bottle;
(4) Placing the penicillin bottle filled with the supernatant solution into a cell disruptor, and ultrasonically controlling the particle size of the liposome by using the cell disruptor again to finally obtain a yellow clarified liposome solution, namely an ROS-responsive quercetin nanoliposome solution;
step 3, adding a mitochondrion targeting molecule into the ROS-responsive quercetin liposome
The steps of adding the mitochondrial targeting molecule to the ROS-responsive quercetin liposomes are as follows:
polyethylene glycol modified triphenyl phosphonic acid-2000 and ROS response type quercetin liposome prepared in the last step are added according to the weight ratio of 2mg: placing 3ml of the mixture into a round-bottom flask, heating the mixture in a water bath at 50 ℃ and continuously stirring the mixture by a magnetic rotor for 2 hours to obtain the ROS-responsive type mitochondria targeted quercetin liposome containing triphenyl phosphate molecules.
2. The method for preparing an ROS-responsive mitochondrial-targeted quercetin liposome according to claim 1, wherein in step 1, when the crude product is purified by gradient elution silica gel column chromatography, the eluent a is dichloromethane: 5 parts of methanol; the eluent B used was dichloromethane: methanol: water 65.
3. The method for preparing ROS-responsive mitochondrial-targeted quercetin liposomes according to claim 1, wherein in the 2 nd step, chloroform and methanol are mixed in a chloroform/methanol mixed solvent at a volume ratio of 3:1.
4. A ROS-responsive mitochondrially targeted quercetin liposome prepared by the method of any one of claims 1 to 3.
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| CN102977360B (en) * | 2012-12-06 | 2014-07-02 | 北京大学 | Triphenylphosphine-polyethyleneglycol 1000 vitamin E succinate (TPGS 1000-TPP) conjugated compound, and preparation method and application thereof |
| UA111762C2 (en) * | 2014-07-08 | 2016-06-10 | ТОВАРИСТВО З ОБМЕЖЕНОЮ ВІДПОВІДАЛЬНІСТЮ "НаноМедТраст" | A method of obtaining a pharmacologically active liposomal drug containing quercetin |
| CN108478534A (en) * | 2018-04-25 | 2018-09-04 | 桂林医学院 | The preparation method and applications of Quercetin liposome |
| CN112402378A (en) * | 2020-11-26 | 2021-02-26 | 烟台大学 | ROS (reactive oxygen species) -responsive dexamethasone liposome containing selenium bond as well as preparation method and application thereof |
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