CN111110863A - Rhein phospholipid complex, preparation method and application thereof, rhein phospholipid complex long-circulating liposome and preparation method thereof - Google Patents

Rhein phospholipid complex, preparation method and application thereof, rhein phospholipid complex long-circulating liposome and preparation method thereof Download PDF

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CN111110863A
CN111110863A CN202010068053.2A CN202010068053A CN111110863A CN 111110863 A CN111110863 A CN 111110863A CN 202010068053 A CN202010068053 A CN 202010068053A CN 111110863 A CN111110863 A CN 111110863A
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rhein
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张金洁
李建波
强弘
张振中
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Zhengzhou University
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Abstract

The invention provides a rhein phospholipid complex and a preparation method and application thereof aiming at the problem of clinical administration of rhein, wherein the preparation method of the rhein phospholipid complex comprises the following steps: (1) weighing rhein and first phospholipid according to the weight ratio of 2: 1-1: 20, and dispersing in an organic solvent to obtain a reaction solution; (2) stirring the reaction solution at 30-60 ℃ for reaction for 2-24 h; (3) concentrating the reacted solution under reduced pressure, and collecting the obtained solid, namely the rhein phospholipid complex; the preparation method provided by the invention is adopted to compound rhein and phospholipid, so that the solubility is improved, the pharmaceutical property is improved, the preparation is facilitated, the long-circulating liposome is further prepared, and the biological half-life period of rhein is effectively prolonged while the high-efficiency drug-loading rate is ensured.

Description

Rhein phospholipid complex, preparation method and application thereof, rhein phospholipid complex long-circulating liposome and preparation method thereof
Technical Field
The invention belongs to the technical field of rhein preparations, and particularly relates to a preparation method of a rhein phospholipid complex, which is further prepared into a long circulating liposome.
Background
Rhein is one of effective components of Chinese medicine rhubarb, belongs to 1,8-Dihydroxy anthraquinone derivatives of monoanthracene nucleus, has the chemical name of 1, 8-Dihydroxy-3-carboxyl anthraquinone (1, 8-Dihydroxy-3-carboxyl-anthraquinone), and has the structural formula as follows:
Figure RE-GDA0002424308000000011
structurally, Rhein contains a planar 1, 8-dihydroxyanthraquinone structural unit, has electrochemical redox property, and endows Rhein with wide Pharmacological effects such as anticancer, antibacterial and anti-inflammatory, immunosuppression and antioxidation and great clinical application value (Sun H, Luo G, Chen D, Xuang Z. aerobic and System review for the pharmaceutical mechanism of Action of Rhein, an Active ingredient of frontiers in pharmacology.2016; 7: 247.). It is worth mentioning that diacetyl rhein (Diacerein) obtained by acetylating two hydroxyl groups of rhein is already marketed in italy as a drug for treating osteoarthritis, and can be rapidly metabolized by the human body into rhein to play a role. However, the special planar structure and the co-planar 1-position hydroxyl, 8-position hydroxyl and 3-position carboxyl cause the compound to have poor water solubility and fat solubility, very poor drug forming property, difficult preparation of the preparation, short biological half-life and serious limitation on the clinical application of the compound.
Due to the defects, the novel delivery system carrying rhein reported in the literature so far is very few, the drug-carrying amount of the obtained delivery system is very low, and no corresponding dosage form is available on the market. Chinese patent CN201811431853 discloses Rhein lipid vesicle nanoparticles with kidney targeting distribution characteristics and application thereof, which are used for drug delivery after Rhein is encapsulated by cationic polymers such as polycaprolactone-polyethyleneimine and the like, and have higher encapsulation efficiency and kidney targeting distribution characteristics on Rhein. However, the cationic polymer nanoparticles prepared by the patent have no long-circulating effect, and the used cationic polymer has strong cytotoxicity and serious side effects such as erythrocyte hemolysis and the like, so the clinical application is greatly limited.
Chinese patent CN105288648A discloses a phospholipid compound of hydrophilic drugs, its pharmaceutical composition and application, wherein a pharmaceutical composition is provided, which uses rhein as active ingredient and phospholipids or cholesterol as adjuvant, but the phospholipid compound prepared by the patent combines drugs and phospholipids by covalent bond, changes the structure of original drug, and needs to release rhein to exert drug effect by breaking chemical bond. In addition, modification of chemical bonds also greatly limits drug loading of phospholipid compounds.
Disclosure of Invention
Aiming at the problems of clinical administration of rhein, the invention firstly provides a rhein phospholipid complex and a preparation method and application thereof.
Meanwhile, the invention also researches that the rhein phospholipid compound is further prepared into the long-circulating liposome and provides a corresponding preparation method.
The invention adopts the following technical scheme:
a preparation method of a rhein phospholipid complex comprises the following steps: (1) weighing rhein and first phospholipid according to the weight ratio of 2: 1-1: 20, and dispersing in an organic solvent to obtain a reaction solution; (2) stirring the reaction solution at 30-60 ℃ for reaction for 2-24 h; (3) and (3) concentrating the reacted solution under reduced pressure, and collecting the obtained solid, namely the rhein phospholipid complex.
Preferably, in the step (1), the amount of the organic solvent is controlled such that the concentration of the rhein in the reaction solution is 1 to 50 mg/ml.
Preferably, in step (1), the organic solvent is one or more of ethyl acetate, acetone, chloroform, dichloromethane, tetrahydrofuran, n-hexane, ethanol and methanol, preferably chloroform or absolute ethanol.
Preferably, the first phospholipid is one or more of lecithin, soybean phospholipid, hydrogenated yolk phospholipid, dipalmitoylphosphatidylethanolamine, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, dimyristoylphosphatidylcholine, dioleoylphosphatidylethanolamine, dipalmitoylphosphatidylglycerol, dipalmitoylphosphatidic acid, phosphatidylcholine and phosphatidylethanolamine. The preferred first phospholipid of the present invention is soybean phospholipid and/or lecithin.
The rhein phospholipid complex is prepared by the preparation method of the rhein phospholipid complex.
The long-circulating lipid nanoparticle prepared from the rhein phospholipid compound comprises 10 parts of the rhein phospholipid compound, 10-50 parts of second phospholipid, 10-50 parts of cholesterol and 1-10 parts of distearoyl phosphatidyl ethanolamine-polyethylene glycol.
Preferably, the molecular weight of the polyethylene glycol in the distearoylphosphatidylethanolamine-polyethylene glycol is 2000-5000-.
Preferably, the second phospholipid is one or more of lecithin, soybean phospholipid, hydrogenated yolk phospholipid, dipalmitoylphosphatidylethanolamine, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, dimyristoylphosphatidylcholine, dioleoylphosphatidylethanolamine, dipalmitoylphosphatidylglycerol, dipalmitoylphosphatidic acid, phosphatidylcholine and phosphatidylethanolamine. The preferred phospholipids of the present invention are soy phospholipids and/or lecithins.
Further, the preparation method of the long-circulating lipid nanoparticles prepared from the rhein phospholipid complex comprises the following steps:
(1) taking the rhein phospholipid complex according to the weight ratio, adding the rhein phospholipid complex into an organic solvent, stirring and dissolving the rhein phospholipid complex into a transparent oily liquid state to obtain a rhein phospholipid complex solution; wherein the organic solvent is chloroform, dichloromethane and other solvents with good dissolving capacity for rhein phospholipid complex, preferably chloroform and dichloromethane;
(2) adding three lipid materials of second phospholipid, cholesterol and distearoyl phosphatidyl ethanolamine-polyethylene glycol which are taken according to the weight ratio into the rhein phospholipid complex solution prepared in the step (1), and stirring until all the lipid materials are completely melted to obtain a clear and transparent oily solution;
(3) concentrating the oily solution prepared in the step (2) under reduced pressure under the conditions of a vacuum degree of 120-150 mbar and a temperature of 25-40 ℃, and removing the organic solvent to form a uniform film;
(4) and (4) adding deionized water into the film formed in the step (3), hydrating the film under water bath ultrasound, and performing ultrasound through a probe to obtain the rhein phospholipid complex long-circulating liposome.
The rhein phospholipid complex can be further added with pharmaceutically acceptable excipient to prepare various pharmaceutically acceptable dosage forms, including but not limited to tablets, capsules, granules, suspensions or powder injections, and the like, and can be applied through the administration routes of gastrointestinal tracts, oral cavities, rectum, skin and the like.
The invention has the following beneficial effects:
phospholipid complexes (phytosomes) are relatively stable compounds or complexes of drugs and phospholipid molecules formed by charge transport. The oxygen atom on the phosphorus atom in the phospholipid structure has a strong tendency of getting electrons, and the nitrogen atom has a strong tendency of losing electrons, so that the phospholipid structure can generate a compound with a drug molecule with a certain structure under certain conditions. After the drug and phospholipid form a complex, the physicochemical properties, biological activity and the like are changed to a great extent, and the complex shows a plurality of characteristics different from those of the parent drug. The change of physicochemical properties such as obviously enhanced fat solubility, obvious change of melting point, absorption coefficient, spectral characteristics and the like, and the change of biological activity such as the activity of the phospholipid complex is generally stronger than that of a parent drug, higher bioavailability and smaller toxic and side effects.
The invention discovers through creative research that oxygen on carboxyl and oxygen on phenolic hydroxyl of rhein both have electronegativity and can generate dipole-dipole acting force with quaternary ammonium nitrogen with electropositivity in lecithin to form a phospholipid complex, so that a planar 1, 8-dihydroxyanthraquinone structural unit of rhein is damaged, the fat solubility and the water solubility of rhein can be obviously improved, and if the rhein phosphate complex is further loaded into liposome, the drug loading capacity of rhein can be greatly improved. Polyethylene glycol modified material can be further added into the liposome material for preparing liposome, which can remarkably prolong the circulation time of rhein in blood, thereby remarkably improving the drug effect of rhein.
For rhein, phospholipid is a proper composite material capable of obviously improving the property of a finished medicine, and is mainly shown in the following aspects that ① phospholipid structure has a functional group capable of being compounded with rhein, ② the rhein phospholipid compound prepared by the invention can break the rigid planar structure of the rhein, improve the fat solubility and the water solubility of the rhein, improve the property of the rhein, can be widely applied to a novel drug delivery system, ③ phospholipid has no obvious toxic or side effect, and ④ the compound method is simple, easy for large-scale production, low in production cost and good in application prospect.
The rhein phospholipid complex prepared by the invention can be used for resisting cancers, inflammation, oxidation, virus infection, headache, coronary heart disease, chronic inflammation and the like, and can be prepared into a rhein phospholipid complex long-circulating liposome.
Drawings
FIG. 1: comparing the results of DSC analysis of rhein phospholipid complex and physical mixture (A. rhein, B. phospholipid, C. complex, D. physical mixture);
FIG. 2: comparison of results of IR analysis of Rhein phospholipid Complex and physical mixture (A. rhein, B. phospholipid, C. Complex; D. physical mixture);
FIG. 3: comparing the XRD analysis results of the rhein phospholipid complex and the physical mixture (A. rhein, B. phospholipid, C. complex, D. physical mixture);
FIG. 4: blood concentration (mean ± SD, n ═ 5) at different times in rats administered with rhein, rhein phospholipid complex common liposomes and rhein phospholipid complex long circulating liposomes intravenously.
Detailed Description
The technical solution of the present invention is further described below with reference to specific examples.
Example 1
Taking 0.3g of rhein and 0.6g of soybean lecithin, adding 50ml of absolute ethanol, stirring and reacting at 45 ℃ for 24h to obtain a clear solution, removing the ethanol at 50 ℃ under reduced pressure, collecting the solid, drying for 12h in vacuum, and crushing to obtain the rhein phospholipid compound.
Dissolving 0.9g of the obtained rhein phospholipid complex in 50mL of chloroform, and stirring and dissolving to obtain a rhein phospholipid complex solution; then, 1.8g of soybean lecithin, 1.2g of cholesterol and 0.18g of distearoylphosphatidylethanolamine-polyethylene glycol were added thereto, and the mixture was stirred until dissolved to form a clear and transparent oily solution. And (2) carrying out reduced pressure rotary evaporation at the temperature of 35 ℃ to remove the solvent to form a film, adding 10mL of deionized water, hydrating under the action of water bath ultrasound (80w for 3min), and then carrying out probe ultrasound (200w for 2min, setting work for 3s, pause for 7s, and total circulation for 12 times) to finally obtain the rhein phospholipid complex long-circulating liposome.
Example 2
Taking 0.2g of rhein and 0.8g of yolk lecithin, adding 40ml of dichloromethane, stirring and refluxing at 40 ℃ for 10h to obtain a clear solution, removing the dichloromethane under reduced pressure at 50 ℃, collecting solids, drying in vacuum for 12h, and crushing to obtain the rhein phospholipid compound.
Dissolving 1g of the obtained rhein phospholipid complex in 50mL of dichloromethane, and stirring and dissolving the obtained rhein phospholipid complex to form a transparent oily liquid to obtain a rhein phospholipid complex solution; then, 1.2g of lecithin, 1g of cholesterol and 0.5g of distearoylphosphatidylethanolamine-polyethylene glycol were added and dissolved to form a clear and transparent oily solution. Performing rotary evaporation at 30 deg.C under reduced pressure to remove solvent to form film, adding deionized water, hydrating under the action of water bath ultrasound (80w,3min), performing probe ultrasound (200w, 2min, setting work for 3s, intermittent for 7s, and circulating for 12 times), and finally obtaining rhein phospholipid complex long-circulating liposome.
Example 3
Taking 0.5g of rhein and 2g of soybean lecithin, adding 50ml of methanol, stirring and reacting at 45 ℃ for 24h to obtain a clear solution, removing ethanol at 50 ℃ under reduced pressure, collecting solids, drying for 12h in vacuum, and crushing to obtain the rhein phospholipid complex.
Dissolving 1.0g of the obtained rhein phospholipid complex in 50mL of chloroform, and stirring to dissolve the rhein phospholipid complex to obtain a rhein phospholipid complex solution; then adding 2.0g lecithin, 1.0g cholesterol and 0.2g distearoyl phosphatidyl ethanolamine-polyethylene glycol, and dissolving to form clear oily solution; performing rotary evaporation at 30 deg.C under reduced pressure to remove solvent to form film, adding 5mL deionized water, hydrating under the action of water bath ultrasound (60w,5min), performing probe ultrasound (200w, 3min, setting work for 3s, intermittent 7s, and circulating for 18 times), and finally obtaining rhein phospholipid complex long-circulating liposome.
Example 4
Adding 0.9g of the rhein phospholipid complex prepared in the example 1 into 30ml of dichloromethane, and stirring and dissolving the mixture until the mixture is in a transparent oily liquid state to obtain a rhein phospholipid complex solution; then adding 1g of soybean lecithin, 0.9g of cholesterol and 0.2g of distearoyl phosphatidyl ethanolamine-polyethylene glycol, stirring and dissolving to completely melt the lipid material, thus obtaining a clear and transparent oily solution. Concentrating under reduced pressure at 35 deg.C, removing low boiling point organic solvent to form uniform film, adding deionized water, hydrating under the action of water bath ultrasound (70w,3min), and performing probe ultrasound (200w, 2min, setting work for 3s, pause for 7s, and circulating for 12 times) to obtain Rhein phospholipid complex liposome.
Comparative example 1
Adding 0.3g of rhein and 0.6g of soybean lecithin into a mortar, and grinding and mixing the rhein and the soybean lecithin uniformly to obtain a physical mixture of the rhein and the lecithin.
Comparative example 2
Taking 0.3g of rhein and 0.6g of soybean lecithin, adding 50ml of absolute ethyl alcohol, and stirring and reacting at 45 ℃ for 24 hours to obtain a clear solution. Removing ethanol at 50 deg.C under reduced pressure, collecting solid, vacuum drying for 12 hr, and pulverizing to obtain Rhein phospholipid complex. Dissolving 0.9g of the obtained rhein phospholipid complex in chloroform, adding 1.8g of soybean lecithin and 1.2g of cholesterol, and stirring until the rhein phospholipid complex is dissolved to form a clear and transparent oily solution. Under the condition that the temperature is 35 ℃, rotary evaporation is carried out to remove the solvent, a film is formed, 10mL deionized water bath ultrasound (80w,3min) is added for hydration, and then probe ultrasound (200w, 2min, setting work for 3s, pause for 7s, and total circulation for 12 times) is carried out to obtain the rhein phospholipid complex common liposome.
DSC analysis of example 1 and comparative example 1
With Al2O3Is a reference substance, and the heating rate is as follows: 10 ℃/min, scan range: 10-400 deg.C, nitrogen flow rate is 0.2ml/min, and rhein, soybean lecithin, rhein phospholipid complex of example 1 and physical mixture of comparative example 1 (10 mg) are respectively analyzed, and the results are shown in figure 1. From DSC, rhein has an obvious endothermic peak, a strong endotherm at 331.12 ℃ shows a peak, and a weak endotherm at about 370 ℃ has a blunt peak. The soybean lecithin has three obvious endothermic peaks, a blunt peak is respectively generated at about 221.9 ℃ and 306.7 ℃, and a sharp peak is generated at about 262.8 ℃. The spectrum of the physical mixture of comparative example 1 exhibited a similar endothermic tendency to that of rhein and phospholipid, and the phase transition temperature range was unclearThe change is significant. In the spectrum of the rhein-phospholipid complex in example 1, the original endothermic peaks of phospholipid and rhein are all lost, and the complex only has weak heat absorption at about 315.4 ℃ and 357 ℃, and has no obvious heat absorption phenomenon. The above results indicate that an interaction occurs between rhein and phospholipids, such as the formation of hydrogen bonds or the presence of van der waals forces, thereby forming phospholipid complexes.
IR analysis of example 1 and comparative example 1
Rhein, soy lecithin, the rhein phospholipid complex of example 1 and the physical mixture of comparative example 1 were scanned in an infrared scan and the results are shown in figure 2. The absorption peak of carboxyl-COOH of rhein is 1714.20cm-1The physical mixture of comparative example 1 and the formation of the rhein phospholipid complex of example 1 showed substantially no change in the peak position, but the peak shape was more or less deactivated, whereas the rhein phospholipid complex of example 1 showed a greater degree of deactivation, indicating that the carboxyl-COOH group of rhein might interact with some groups of phospholipid under both conditions of comparative example 1 and example 1, but the intensity of the interaction was different. The C ═ O absorption peak of the phospholipid itself was 1744.56 cm-1The absorption peak of P ═ O is 1245.73cm-1In the spectrum of the physical mixture of the comparative example 1, the peak positions of the two are basically unchanged, but the peak shapes are slightly passivated, while in the spectrum of the rhein phospholipid complex of the example 1, the C ═ O absorption peak is shifted to 1735.62 cm-1It is evident that P ═ O absorption is also greatly diminished, presumably because the phenolic-OH of rhein associates to some extent with P ═ O and C ═ O of phospholipids. The above changes in peak position and peak shape indicate that the rhein phospholipid complex of example 1 has an infrared spectrum different from that of a physical mixture thereof and exists as a complex.
XRD analysis of example 1 and comparative example 1
XRD analysis was performed on rhein, soybean lecithin, rhein phospholipid complex and the physical mixture of comparative example 1, and the results are shown in FIG. 3. As can be seen from the graphs, the four samples show different diffraction characteristics, and the graph of the physical mixture of the comparative example 1 still has obvious peak diffraction characteristics and no newly generated peak compared with rhein; the rhein phospholipid complex pattern of example 1 shows an amorphous structure characteristic similar to that of phospholipid, and the crystal diffraction peak of rhein completely disappears. The above shows that the rhein phospholipid complex of example 1 is obviously different from a simple physical mixture, and after the phospholipid complex is formed, rhein and phospholipid are in a highly dispersed state due to certain interaction, namely directional combination, of certain groups in the structure of rhein and polar ends of phospholipid, and the particle structure and the crystal structure of the rhein and the phospholipid are damaged.
Examples of pharmacokinetic experiments
The experiment examines rhein, the rhein phospholipid complex common liposome prepared in the comparative example 2 and the distearoylphosphatidylethanolamine-polyethylene glycol modified rhein phospholipid complex liposome prepared in the example 1 have bioavailability in rats
1. Materials and animals:
rhein phospholipid Complex (prepared according to example 1), Rhein phospholipid Complex Normal liposomes (prepared according to comparative example 2), Distearylphosphatidylethanolamine-polyethylene glycol modified Rhein phospholipid Complex Long-circulating liposomes (prepared according to example 1)
Rhein: purchased from Sigma-aldrich with a purity > 96% (HPLC)
Male Wistar rat 15 (weight 200 ~ 300g)
2. The method comprises the following steps:
the administration scheme is as follows: 15 Wistar rats were randomly divided into three groups, and after fasting for 12 hours, the three groups were each intravenously administered with rhein phospholipid complex (prepared according to example 1), rhein phospholipid complex ordinary liposome (prepared according to comparative example 2), distearoylphosphatidylethanolamine-polyethylene glycol-modified rhein phospholipid complex long-circulating liposome (prepared according to example 1)
The administration dosage is equivalent to 10mg/kg of rhein. Collecting samples: after administration of each group, 0.3ml of blood was collected from the tail vein at 5, 10, 30, 60, 120, 240, 360, 480min, placed in a heparin-treated centrifuge tube, plasma was centrifuged and stored in a refrigerator at-20 ℃ until analysis. Plasma sample treatment: and taking 100 mul of plasma sample, adding the plasma sample into a clean centrifuge tube, adding 300 mul of methanol, mixing uniformly by vortex for 5min, centrifuging for 15min at 14000rpm, and taking 100 mul of supernatant for sample injection detection.
Chromatographic conditions are as follows: a chromatographic column: Kromasil-C18 column (150 mm. times.4.6 mm,5 μm); mobile phase: methanol-0.1% phosphoric acid water by volume (85:15, v/v); flow rate: 1 ml/min; detection wavelength: 258 nm; column temperature: 25 ℃; sample introduction amount: 20 μ l.
The method comprises the following steps: the linear relationship is examined by rhein, and the rhein linear range is as follows: 0.48-48 μ g/ml (r ═ 0.9998). Exclusive examination shows that endogenous substances in blood plasma do not interfere with the determination result, and the separation degree of the internal standard and the standard is good. The recovery rates of the high, medium and low concentrations of rhein are respectively 98.83%, 100.7% and 98.7%, and the RSD in the day and the RSD in the daytime are respectively 2.2%, 5.1% and 3.7%; 2.6%, 4.3%, 3.1%.
3. Results
The results of the blood concentration measurement are shown in fig. 4.
Calculation of pharmacokinetic parameters:
plasma concentration-time data of rhein phospholipid complex (prepared according to example 1), rhein phospholipid complex common liposome (prepared according to comparative example 2), distearoylphosphatidylethanolamine-polyethylene glycol modified rhein phospholipid complex long-circulating liposome (prepared according to example 1) were subjected to computer fitting using DAS 3.0 software to obtain AUC and related parameters, and the main pharmacokinetic parameters are shown in table 1:
TABLE 1 pharmacokinetic parameters of Rhein, Rhein phospholipid Complex Normal liposomes and Rhein phospholipid Complex Long circulating liposomes (equivalent to Rhein dose 2.5mg/kg) administered in tail vein of rats (n ═ 5)
Figure BDA0002376548070000081
Substituting the above main drugsThe kinetic parameters were subjected to a t-test (in which,*p<0.05, compared to the rhein group;#p<0.05, compared to the rhein phospholipid complex common liposome group), the results showed AUC of the rhein phospholipid complex long-circulating liposome group(0-t)(pg/L) × h and MRT(0-t)(h) Is significantly higher than rhein group (*p<0.05), which shows that the rhein phospholipid complex long-circulating liposome can obviously improve the blood concentration of rhein and obviously prolong the in-vivo circulation time of rhein. In addition, t of Rhein phospholipid Complex Long circulating Liposome group1/2(h) Also significantly higher than rhein group (*p<0.05), which shows that the rhein can remarkably reduce the clearance rate of rhein in vivo. Meanwhile, the common liposome is also found to be capable of obviously increasing the drug concentration in the blood plasma of the retinoic acid and prolonging the circulation time in vivo after the common liposome is injected into the vein of a rat. The half-life period in vivo of the three administration groups is in the order of rhein phospholipid complex long-circulating liposome group>Group of common liposomes>Rhein group. Therefore, the rhein phospholipid complex common liposome and the rhein phospholipid complex long-circulating liposome can obviously improve the half-life period of rhein in vivo, wherein the rhein phospholipid complex long-circulating liposome group has better improvement effect than the rhein phospholipid complex common liposome, which is probably related to that DSPE-PEG can inhibit the effect of the liposome and opsonin in plasma, thereby prolonging the in vivo circulation time of the liposome.
Finally, it should be noted that: the above embodiments are merely illustrative and not restrictive of the technical solutions of the present invention, and any equivalent substitutions and modifications or partial substitutions made without departing from the spirit and scope of the present invention should be included in the scope of the claims of the present invention.

Claims (9)

1. A preparation method of a rhein phospholipid complex is characterized by comprising the following steps: (1) weighing rhein and first phospholipid according to the weight ratio of 2: 1-1: 20, and dispersing in an organic solvent to obtain a reaction solution; (2) stirring the reaction solution at 30-60 ℃ for reaction for 2-24 h; (3) and (3) concentrating the reacted solution under reduced pressure, and collecting the obtained solid, namely the rhein phospholipid complex.
2. The method for preparing the rhein phospholipid complex according to claim 1, wherein in the step (1), the amount of the organic solvent is controlled to be 1-50 mg/ml of rhein in the reaction solution.
3. The method for preparing the rhein phospholipid complex according to claim 1, wherein in the step (1), the organic solvent is one or more of ethyl acetate, acetone, chloroform, dichloromethane, tetrahydrofuran, n-hexane, ethanol and methanol.
4. The method for preparing the rhein phospholipid complex, according to claim 1, wherein the first phospholipid is one or more selected from lecithin, soybean phospholipid, hydrogenated egg yolk phospholipid, dipalmitoylphosphatidylethanolamine, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, dimyristoylphosphatidylcholine, dioleoylphosphatidylethanolamine, dipalmitoylphosphatidylglycerol, dipalmitoylphosphatidic acid, phosphatidylcholine, and phosphatidylethanolamine.
5. The rhein phospholipid complex prepared by the method for preparing the rhein phospholipid complex according to the claims 1 to 5.
6. The long-circulating lipid nanoparticle prepared from the rhein phospholipid complex of claim 6 is characterized by comprising 10 parts of the rhein phospholipid complex, 10-50 parts of second phospholipid, 10-50 parts of cholesterol and 1-10 parts of distearoyl phosphatidyl ethanolamine-polyethylene glycol in parts by weight.
7. The long-circulating lipid nanoparticle prepared from the rhein phospholipid complex of claim 7, wherein the molecular weight of the polyethylene glycol in the distearoylphosphatidylethanolamine-polyethylene glycol is 2000-5000-.
8. The rhein phospholipid complex prepared long-circulating lipid nanoparticle according to claim 7, wherein the second phospholipid is one or more of lecithin, soybean phospholipid, hydrogenated egg yolk phospholipid, dipalmitoylphosphatidylethanolamine, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, dimyristoylphosphatidylcholine, dioleoylphosphatidylethanolamine, dipalmitoylphosphatidylglycerol, dipalmitoylphosphatidic acid, phosphatidylcholine, and phosphatidylethanolamine.
9. The method for preparing the long-circulating lipid nanoparticles prepared from the rhein phospholipid complex according to any one of claims 7 to 9, which is characterized by comprising the following steps:
(1) taking the rhein phospholipid complex according to the weight ratio, adding the rhein phospholipid complex into an organic solvent, stirring and dissolving the rhein phospholipid complex into a transparent oily liquid state to obtain a rhein phospholipid complex solution;
(2) adding three lipid materials of second phospholipid, cholesterol and distearoyl phosphatidyl ethanolamine-polyethylene glycol which are taken according to the weight ratio into the rhein phospholipid complex solution prepared in the step (1), and stirring until all the lipid materials are completely melted to obtain a clear and transparent oily solution;
(3) concentrating the oily solution prepared in the step (2) under reduced pressure under the conditions of a vacuum degree of 120-150 mbar and a temperature of 30-45 ℃, and removing the organic solvent to form a uniform film;
(4) and (4) adding deionized water into the film formed in the step (3), hydrating the film under water bath ultrasound, and performing ultrasound through a probe to obtain the rhein phospholipid complex long-circulating liposome.
CN202010068053.2A 2020-01-21 2020-01-21 Rhein phospholipid complex, preparation method and application thereof, rhein phospholipid complex long-circulating liposome and preparation method thereof Pending CN111110863A (en)

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