CN115919775A - A kind of fat-soluble drug lipid composition and its preparation method and application - Google Patents

Abstract

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a fat-soluble pharmaceutical lipid composition, and a preparation method and application thereof. The fat-soluble drug lipid composition provided by the invention comprises a drug-loaded liposome and a dispersion solvent; the drug-loaded liposome comprises a lipid material and a fat-soluble drug, wherein the fat-soluble drug comprises one or more of cepharanthine, cepharanthine salt, cepharanthine derivatives, ivermectin salt and ivermectin derivatives, and the lipid material comprises one or more of phosphatidyl choline, phosphatidyl ethanolamine, phosphatidyl glycerol, acyl phosphatidic acid, phosphatidyl serine and lysophospholipid; the mass concentration of the lipid material in the dispersing solvent is 10-300 mg/mL; in the fat-soluble medicine lipid composition, the medicine-loading rate of the fat-soluble medicine is more than or equal to 8mg/mL. The fat-soluble medicine lipid composition can realize high drug loading rate and high medicine encapsulation rate of the cepharanthine and/or ivermectin, and has wide application prospect.

Description

A kind of fat-soluble drug lipid composition and its preparation method and application

technical field

The invention belongs to the technical field of pharmaceutical preparations, and in particular relates to a fat-soluble pharmaceutical lipid composition and its preparation method and application.

Background technique

CEP is a bisbenzylisoquinoline alkaloid monomer compound isolated from the root tubers of Stephaniaceae plants of the family Fangjiaceae, Stephania, Dirong and Jinxiandang tortoise. The molecular formula of stephadin is C ₃₇ H ₃₈ N ₂ O ₆ , and the molecular weight is 606.71. It is light yellow amorphous powder, odorless, with bitter taste, darkens when exposed to light, and has the property of sublimation.

Modern pharmacological studies have shown that stephatrin has anti-virus, anti-tumor, immune regulation, increased white blood cells, CEP and chloroquine synergistic anti-malarial, treatment of endotoxic shock, treatment of recurrent oral ulcers, and primary thrombocytopenic purpura And so on. In my country, it is mainly used for the prevention of silicosis and pneumoconiosis and the treatment of leukopenia caused by radiotherapy and chemotherapy. Stephaniacine has a clinical application history of more than 40 years, and its safety has been fully verified. In particular, in the context of the raging novel coronavirus, the development of therapeutic drugs is urgently needed. Stephalin has shown good anti-coronavirus effects and has attracted widespread attention from medical workers, and may become a very promising therapeutic drug.

Throughout the domestic and foreign use of stephatrin-related preparations, the marketed dosage forms are only tablets and injections, and the drug loading is relatively low. The dosage is 0.5% (W/V, equivalent to 5mg/mL). And it is reported in the literature that the drug-loading capacity of stepherin lotion is 0.1% (W/V, equivalent to 1mg/mL), and the drug-loading capacity of stepherin nanostructured lipid carrier is 1% (W/W); Chinese patent CN102475680A discloses "a kind of stepherin hydrochloride injection and its preparation method and detection method". and its preparation method", although the drug-loading capacity of stephatrin nanosuspension can reach 10.3% (W/V), but stephatrin exists in the form of solid nanocrystals, compared with stephatrin that exists in molecular state The hormone absorption is relatively poor, and the stability is poor, easy to aggregate, and the route of administration is single. At the same time, when taking orally administered as a tablet, stepherin has a serious hepatic first-pass effect, and its bioavailability is low; because stepherin metabolizes faster in the body, its elimination half-life is shorter (T _1/2 is 4.62h), therefore, Even if the conventional injection preparation prepared by solubilization is still difficult to maintain a stable blood concentration, in addition, due to frequent injections and the pain caused by the injection, the patient's medication compliance decreases accordingly. Therefore, the dosage forms corresponding to the above two routes of administration of stephatrin have relatively low drug loading, which directly affects the efficacy of the drug, resulting in poor therapeutic effect.

Ivermectin is a new type of broad-spectrum, high-efficiency, and low-toxicity antibiotic antiparasitic drug, which has a good killing effect on internal and external parasites, especially nematodes and arthropods. Ivermectin is a white crystalline powder; odorless. This product is soluble in methanol, ethanol, acetone, ethyl acetate, almost insoluble in water, slightly hygroscopic. Ivermectin is a sixteen-membered ring macrolide antibiotic, its basic structure is a large ester ring at carbon 16 and three main substituent groups, namely hexahydrophenylpropanoid at C2 to C8 Furans, disaccharide group at C13, keto group at C17 to C18.

Ivermectin is an older drug approved by the FDA for parasitic infections. In vitro cell culture experiments have shown that ivermectin can effectively inhibit the new coronavirus (SARS-CoV-2) in vitro, and a single administration can reduce the viral RNA by about 5000 times within 48 hours, so it has the ability to fight new coronaviruses with old drugs and new applications potential. However, the concentration of ivermectin used in in vitro experiments is difficult to achieve in human lungs or plasma. Even if the oral dose of ivermectin is increased to 10 times its antiparasitic dose (200 μg/kg), the maximum The plasma concentration (0.3μM) also cannot reach its anti-new coronavirus IC50 level (2μM).

To sum up, the existing preparations of stephatrin and ivermectin have the problem of low drug loading, which cannot meet the clinical application.

Contents of the invention

The purpose of the present invention is to provide a fat-soluble drug lipid composition and its preparation method and application. The fat-soluble drug lipid composition provided by the present invention can realize high drug loading and high drug loading of stepherin and ivermectin. Encapsulation efficiency, broad application prospects.

In order to achieve the above object, the present invention provides the following technical solutions:

The invention provides a fat-soluble drug lipid composition, including drug-loaded liposomes and a dispersion solvent; , one or more of ivermectin salts and ivermectin derivatives, the lipid materials include phosphatidylcholine compounds, phosphatidylethanolamine compounds, phosphatidylglycerol compounds, acylphosphatidic acids One or more of compounds, phosphatidylserine compounds, and lysophospholipid compounds; the mass concentration of the lipid material in the dispersion solvent is 10-300 mg/mL;

In the fat-soluble drug lipid composition, the drug loading amount of the fat-soluble drug is more than or equal to 8 mg/mL.

Preferably, the lipid material includes hydrogenated soybean lecithin, egg yolk lecithin, dioleoyl lecithin, dipalmitoyl lecithin, distearoyl phosphatidylcholine, 2-palmitoyl phosphatidylglycerol, distearyl One or more of acylphosphatidylethanolamine-polyethylene glycol.

Preferably, the lipid material comprises egg yolk lecithin, dioleoyl lecithin, dipalmitoyl lecithin (DPPC), distearoyl phosphatidylcholine, 2-palmitoyl phosphatidylglycerol, distearoyl phosphatidyl phosphatidylcholine One or more of ethanolamine-polyethylene glycol.

Preferably, the drug loading amount of the fat-soluble drug is 8-20 mg/mL.

Preferably, the dispersion solvent includes one or more of water, physiological saline, glucose solution or phosphate buffered saline solution.

Preferably, the fat-soluble drug lipid composition further includes cholesterol; the mass ratio of the cholesterol to the fat-soluble drug is (0-1):(0.5-2).

The present invention provides the preparation method of the fat-soluble drug lipid composition described in the above technical scheme, including film dispersion method, reverse evaporation method, solvent injection method, freeze-drying method, French pressure method, double emulsion method, high-pressure homogenization method , pH gradient method, microfluidic method and any one of microfluidic method.

The present invention provides the application of the fat-soluble drug lipid composition described in the above technical scheme or the fat-soluble drug lipid composition prepared by the preparation method described in the above technical scheme in the preparation of medicines, and the medicine includes the treatment of leukocytes Drugs for hypothalamus, drugs for silicosis, drugs for pneumoconiosis, drugs for snake bites, drugs for alopecia areata, drugs for hair loss, drugs for essential thrombocytopenic purpura, drugs for mouth ulcers, treatments Drugs for endotoxic shock, anti-malarial drugs, immunomodulatory drugs, antiviral drugs or antineoplastic drugs.

Preferably, the dosage form of the drug includes oral preparations, external preparations or injection preparations.

Preferably, the external preparations include transdermal preparations, ophthalmic preparations or nasal cavity preparations.

The present invention provides a fat-soluble drug lipid composition, comprising a drug-loaded liposome and a dispersion solvent; the drug-loaded liposome includes a fat-soluble drug and a lipid material carrying the fat-soluble drug, the Fat-soluble drugs include stepherin and its salts and/or ivermectin and its salts, and the lipid materials include phosphatidylcholines, phosphatidylethanolamines, phosphatidylglycerols, acylphosphatidic acids, phosphatidyl One or more of serines and lysophospholipids; the mass concentration of the lipid material in the dispersion solvent is 10-300 mg/mL; in the fat-soluble drug lipid composition, the fat-soluble The drug loading amount is ≥8mg/mL. The present invention uses lipid materials to entrap stepherin and its salt and/or ivermectin and its salt in a dispersion solvent to form a dispersion of drug-loaded liposomes, with phosphatidylcholines, phosphatidylethanolamine One or more of lipids, phosphatidylglycerols, acylphosphatidic acids, phosphatidylserines, and lysophospholipids are used as lipid materials, and the mass concentration of lipid materials in the dispersion solvent is controlled at 10-300mg /mL, the fat-soluble drug lipid composition obtained can realize the high drug loading capacity and high drug encapsulation efficiency of stepherin and/or ivermectin, and has broad application prospects.

Description of drawings

Fig. 1 is the particle size distribution figure of the stepherin lipid composition prepared in Example 1 of the present invention;

Fig. 2 is a potential distribution diagram of the stepherin lipid composition prepared in Example 1 of the present invention.

Detailed ways

The present invention provides a fat-soluble drug lipid composition, comprising a drug-loaded liposome and a dispersion solvent; the drug-loaded liposome includes a fat-soluble drug and a lipid material carrying the fat-soluble drug, the The fat-soluble drug includes one or more of stepherin, stepherin salt, stepherin derivative, ivermectin, ivermectin salt and ivermectin derivative, and the lipid material includes One or more of phosphatidylcholine compounds, phosphatidylethanolamine compounds, phosphatidylglycerol compounds, acylphosphatidic acid compounds, phosphatidylserine compounds, lysophospholipid compounds; the lipid material in the The mass concentration in the dispersing solvent is 10-300mg/mL;

In the fat-soluble drug lipid composition, the drug loading amount of the fat-soluble drug is more than or equal to 8 mg/mL.

In the present invention, unless otherwise specified, all preparation raw materials/components are commercially available products well known to those skilled in the art.

The fat-soluble drug lipid composition provided by the present invention includes a drug-loaded liposome, and the drug-loaded liposome includes a fat-soluble drug and a lipid material encapsulating the fat-soluble drug.

In the present invention, the fat-soluble drug includes one or more of stepherin, stepherin salt, stepherin derivative, ivermectin, ivermectin salt and ivermectin derivative .

In the present invention, the stepherine is also known as stepherine, stepherine, and stepherine.

As a specific embodiment of the present invention, the stephatrin is one or more of the extracts of Fenfangcai extract, Stephania vine extract, Dirong extract, and Jinxiandian tortoise extract.

In the present invention, the golden-threaded tortoise is aliased as Baiyao, golden-threaded toad, and cypress.

In the present invention, the drug loading amount of the fat-soluble drug is ≥8 mg/mL, preferably 8-20 mg/mL, more preferably 8.5-19.5 mg/mL.

In the present invention, the drug loading amount of the fat-soluble drug specifically refers to the mass concentration of the fat-soluble drug in the lipid composition of the fat-soluble drug, and the unit is mg/mL.

In the present invention, the mass percentage of the fat-soluble drug in the total solid matter in the fat-soluble drug lipid composition is preferably ≥ 6%, more preferably 7-12%.

In the present invention, the particle size of the drug-loaded liposome in the fat-soluble drug lipid composition is preferably 76-96nm; Zeta potential 1.9-8.5mV; encapsulation efficiency 98.17-99.97%.

In the present invention, the lipid material includes phosphatidylcholine compounds (PCs), phosphatidylethanolamines (PEs), phosphatidylglycerols (PGs), acylphosphatidic acids (PAs) ), phosphatidylserine compounds (PS class), one or more of lysophospholipid compounds; preferably include hydrogenated soybean lecithin (HSPC), egg yolk lecithin (EPC), dioleoyl lecithin (DOPC), Dipalmitoyl lecithin (DPPC), distearoylphosphatidylcholine (DSPC), 2-palmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-MPEG2000) One or more of, more preferably egg yolk lecithin (EPC), dioleoyl lecithin (DOPC), dipalmitoyl lecithin (DPPC), distearoyl phosphatidylcholine (DSPC), 2-palm One or more of acylphosphatidylglycerol (DPPG), distearoylphosphatidylethanolamine-polyethylene glycol (DSPE-MPEG2000).

As a specific embodiment of the present invention, the lipid material is DOPC and DSPE-MPEG2000; the mass ratio of DOPC and DSPE-MPEG2000 is 9:1.

As a specific embodiment of the present invention, the lipid material is DOPC and DPPC; the mass ratio of DOPC and DPPC is 1:1.

As a specific embodiment of the present invention, the lipid material is DOPC and DSPE-MPEG2000; the mass ratio of DOPC and DSPE-MPEG2000 is 135:15.

In the present invention, the total mass concentration of the lipid material in the dispersion solvent is 10-300 mg/mL, preferably 15-280 mg/mL, more preferably 20-265 mg/mL.

In the present invention, when the mass concentration of the lipid material in the dispersion solvent is lower than the above-mentioned range, the ability of the lipid material to entrap the fat-soluble drug is reduced; the effective effect on the fat-soluble drug cannot be realized. Encapsulation; when the mass concentration of the lipid material in the dispersion solvent is higher than the above range, the viscosity of the fat-soluble drug lipid composition system is too high, the fluidity is poor, and the effective delivery of the drug cannot be achieved.

The fat-soluble drug lipid composition provided by the invention includes a dispersion solvent.

In the present invention, the dispersion solvent preferably includes one or more of water, physiological saline, glucose solution or phosphate buffered saline solution. The water is preferably deionized water.

In the present invention, the dispersion solvent more preferably includes water or phosphate buffered saline (PBS). In the present invention, the pH value of the PBS is preferably 7.4.

In the present invention, the dispersion solvent is also called a hydration solution.

In the present invention, the fat-soluble pharmaceutical lipid composition preferably further includes cholesterol.

In the present invention, there is a competitive relationship between the cholesterol and the fat-soluble drug, and they co-exist in the gaps in the phospholipid bilayer formed by the lipid material. In the present invention, cholesterol is preferably added to reduce the density of the phospholipid bilayer. Permeability to prevent leakage of fat-soluble drugs.

In the present invention, when the lipid composition of the fat-soluble drug preferably also includes cholesterol, the mass ratio of the fat-soluble drug to the cholesterol is preferably (0～1):(0.5～2), more preferably (0.1 ~1): (0.6~1.5).

The present invention provides the preparation method of the fat-soluble drug lipid composition described in the above technical scheme, including film dispersion method, reverse evaporation method, solvent injection method, freeze-drying method, French pressure method, double emulsion method, high-pressure homogenization method One or more of pH gradient method, microfluidic method and microfluidic method.

In the present invention, the solvent injection method is preferably an ethanol injection method.

In the present invention, the preparation method of the solvent injection method preferably includes the following steps: dissolving the lipid material and the fat-soluble drug in an organic solvent to obtain a fat-soluble drug solution; injecting the fat-soluble drug solution into the In the dispersion solvent, a mixed solution is obtained; after removing the organic solvent, the mixed solution is ultrasonically dispersed to obtain the fat-soluble drug lipid composition. In the present invention, the organic solvent is preferably ethanol; the ratio of the mass of the lipid material, the mass of the fat-soluble drug and the volume of the solvent is preferably 100mg:10mg:10mL. The volume ratio of the organic solvent and the dispersion solvent is preferably 1:1. The injection is preferably performed using an autonomous syringe pump, and the injection speed is preferably 200 μL/min. In the present invention, the temperature for removing the organic solvent is preferably 55°C. The ultrasound is preferably performed in a cell disruptor, and the power of the ultrasound is preferably 30W-100W, preferably 45-70W. The ultrasonic time is preferably 2-10 min, more preferably 3-6 min. The ultrasound is preferably in an intermittent mode, and the intermittent mode is preferably that the ultrasound lasts for 2s and pauses for 2s. When the ultrasonic time of the present invention is more than 1 min, the mixed solution from which the organic solvent is removed changes from milky white to clear and transparent.

In the present invention, the microfluidic method preferably includes the following steps: dissolving the lipid material, cholesterol and fat-soluble drug in an organic solvent to obtain a fat-soluble drug solution; removing the solvent from the fat-soluble drug solution to obtain lipid film; the lipid film, dispersion solvent and glass beads are mixed for hydration to obtain a hydration solution; the hydration solution is dispersed to obtain the fat-soluble drug lipid composition, which is dispersed in a high-pressure micro-jet carried out in the instrument. In the present invention, the organic solvent is preferably absolute ethanol; the dissolution is preferably carried out under ultrasonic conditions, the ultrasonic power is preferably 220W, and the ultrasonic wave is 45Hz; the ultrasonic dissolution time for 5min. In the present invention, the specific embodiment of the solvent removal is preferably vacuum rotary evaporation, and the temperature of the vacuum rotary evaporation is preferably a 45°C water bath. The dispersing solvent is preferably deionized water. The temperature of the hydration is preferably 50° C., the time of the hydration is preferably 30 minutes, and the water is preferably stirred under the condition of stirring, and the stirring speed is preferably 75 rpm.

In the present invention, the film dispersion method preferably includes the following steps: dissolving the lipid material and the fat-soluble drug in an organic solvent to obtain a fat-soluble drug solution; coating the fat-soluble drug solution to obtain a lipid-soluble drug solution. Film: mixing the lipid film with a dispersion solvent for hydration and then dispersing to obtain the fat-soluble drug lipid composition. In the present invention, the organic solvent is preferably chloroform; the dissolution is preferably performed under ultrasonic conditions. In the present invention, the film laying is preferably rotary film laying, and the lipid film is preferably stored in a vacuum drying oven. The dispersing solvent is preferably PBS. The temperature of the hydration is preferably 55° C., the time of the hydration is preferably 45 minutes, the hydration is preferably carried out under the condition of stirring, and the rotational speed of the stirring is preferably 100 rpm. The dispersion is preferably carried out in a cell disruptor, the time of the ultrasound is 2 minutes, the power of the ultrasound is preferably 45W, and the ultrasound is preferably in an intermittent working mode, which lasts for 2s and pauses for 2s.

In the present invention, the freeze-drying method preferably includes the following steps: dissolving the lipid material and the fat-soluble drug in an organic solvent to obtain a fat-soluble drug solution; dissolving trehalose or mannose in physiological saline to obtain Protecting solution; filtering the fat-soluble drug solution and protecting solution to obtain a filtrate; pre-freezing the filtrate and freeze-drying to obtain the fat-soluble drug lipid composition. The dissolution is preferably performed under ultrasonic conditions. In the present invention, the filtration is preferably performed by using a microporous membrane; the temperature of the pre-freezing is preferably -80° C., and the time of the pre-freezing is preferably 12 hours. The freeze-drying is preferably carried out in a freeze-drying agent, and the freeze-drying time is preferably 24 hours.

In the present invention, the high-pressure homogenization method preferably includes the following steps: dissolving the lipid material and the fat-soluble drug in an organic solvent to obtain a fat-soluble drug solution; coating the fat-soluble drug solution to obtain a lipid-soluble drug solution. a thin film; the lipid film is mixed with a dispersion solvent, homogenized and then freeze-dried to obtain the fat-soluble drug lipid composition. In the present invention, the organic solvent is preferably chloroform; the dissolution is preferably performed under ultrasonic conditions. In the present invention, the film laying is preferably rotary film laying, the film spreading is preferably carried out on a rotary steamer, the temperature of the rotary film laying is preferably 45°C, and the rotating speed of the rotary film laying is preferably 70rpm, The time for the rotary coating is preferably 50 min; the lipid film is preferably stored in a vacuum oven. The homogenization is preferably carried out in a high-pressure homogenizer, and the homogenization preferably includes the following steps: mixing 5 times at 50,000 kPa, and then mixing 3 times at 100,000 kPa. The fat-soluble drug lipid composition is preferably stored at 4°C.

In the present invention, the pH gradient method is preferably an ammonium sulfate gradient method. In the present invention, the ammonium sulfate gradient method preferably includes the following steps: dissolving the lipid material in an organic solvent to obtain a lipid material solution; coating the lipid material solution to obtain a blank film of lipid material ; the lipid material blank film is hydrated with an ammonium sulfate aqueous solution and mixed with a dispersion solvent to obtain a blank lipid; after dialyzing the blank lipid, add a fat-soluble drug to incubate and load the drug to obtain the fat-soluble drug lipid quality composition. In the present invention, the organic solvent is preferably chloroform; the dissolution is preferably performed under ultrasonic conditions. In the present invention, the film laying is preferably rotary film laying, the film spreading is preferably carried out on a rotary steamer, the temperature of the rotary film laying is preferably 45°C, and the rotating speed of the rotary film laying is preferably 70rpm, The time for the rotary coating is preferably 45 min; the molar concentration of the ammonium sulfate aqueous solution is preferably 530 mmol/L. The dialysis is preferably carried out in an aqueous sodium chloride solution, the molar concentration of the aqueous sodium chloride solution is preferably 160mmol/L, the dialysis is preferably carried out at room temperature, and the number of times of the dialysis is preferably 3 times, each dialysis The time is preferably 1h. In the present invention, the temperature for incubating drug loading is preferably 40°C.

The present invention provides the application of the fat-soluble drug lipid composition described in the above technical scheme or the fat-soluble drug lipid composition prepared by the preparation method described in the above technical scheme in the preparation of medicines, and the medicine includes the treatment of leukocytes Drugs for hypothalamus, drugs for silicosis, drugs for pneumoconiosis, drugs for snake bites, drugs for alopecia areata, drugs for hair loss, drugs for essential thrombocytopenic purpura, drugs for mouth ulcers, treatments Drugs for endotoxic shock, anti-malarial drugs, immunomodulatory drugs, antiviral drugs or antineoplastic drugs.

In the present invention, the antiviral drugs preferably include anti-new coronavirus drugs.

In the present invention, the dosage form of the drug preferably includes oral preparations, external preparations or injection preparations.

In the present invention, the external preparation preferably includes a transdermal preparation, an ophthalmic preparation or a nasal administration preparation.

In order to further illustrate the present invention, the technical solutions provided by the present invention will be described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the protection scope of the present invention.

Example 1

The mass ratio relationship of the raw materials of the fat-soluble drug lipid composition provided in this embodiment is shown in Table 1:

Table 1 The raw material mass ratio relationship of the fat-soluble drug lipid composition of this embodiment

In this embodiment, the ethanol injection method is adopted to prepare a high-load stepherin lipid composition, and the specific preparation method includes the following steps:

Weighed the lipid material and stepherin in the prescribed amount in Table 1 and dissolved them in 10 mL of ethanol to obtain a mixed solution, and injected the mixed solution into PBS buffer (pH 7.4) at a constant speed with an automatic syringe pump at 200 μL/min. Heat at a constant temperature of 55°C and continue to stir until the ethanol is completely volatilized. Use a cell disruptor to sonicate for 10 minutes. The ultrasonic power is 45W. Continuously sonicate for 2s and pause for 2s to obtain a colorless and translucent lipid composition with high loading of stepherin. The liposome particle size in the prepared high-load stepherin lipid composition is 84nm, the Zeta potential is 6.5mV, and the drug loading is 10.00mg/mL (stepherin accounts for 10.00mg/mL of the fat-soluble drug lipid composition). Mass concentration), Stephalin accounted for 8.7% by mass of the total solid matter in the fat-soluble drug lipid composition, and the encapsulation efficiency was 99.96%.

Fig. 1 is the particle size distribution diagram of the high-loaded stepherin lipid composition in Example 1 of the present invention. Fig. 2 is a graph showing the potential distribution of the high-load stepherin lipid composition of the present invention in Example 1. From Figure 1 and Figure 2, it can be seen that the liposome particle size distribution in the high-load stepherin lipid composition prepared in this example is narrow, and the liposome particles are uniform.

Example 2

The ratio relationship of the raw materials of the fat-soluble drug lipid composition provided by this embodiment is shown in Table 2:

Table 2 The raw material mass ratio relationship of the fat-soluble drug lipid composition of this embodiment

In this embodiment, a microfluidic method is adopted to prepare a high-capacity stepherin lipid composition, and the specific preparation method includes the following steps:

Weigh the lipid material, cholesterol, and stepherin in the prescribed amount in Table 2, add 500mL of absolute ethanol, ultrasonicate in a water bath at 45Hz and 220W for 5 minutes until fully dissolved, and spin evaporate under reduced pressure in a water bath at 45°C to remove ethanol. After a layer of lipid film is formed, add deionized water and glass beads, rotate and hydrate at 75 rpm for 30 minutes in a water bath at 50°C, and disperse the liquid in a high-pressure micro-fluidic instrument. The dispersing pressure is 1200 bar, and the number of dispersing cycles 7 times, a colorless and translucent high-loading stepherin lipid composition was obtained. The liposome particle size in the prepared high-load stepherin lipid composition is 86nm, the Zeta potential is 3.2mV, and the drug loading is 8.49mg/mL (stepherin accounts for 8.49mg/mL of the fat-soluble drug lipid composition). mass concentration), stepherin accounts for 7.6% by mass of the total solid matter in the fat-soluble drug lipid composition, and the encapsulation efficiency is 99.92%.

Example 3

The ratio relationship of the raw materials of the fat-soluble drug lipid composition provided by this embodiment is shown in Table 3:

Table 3 The raw material mass ratio relationship of the fat-soluble drug lipid composition of this embodiment

In this embodiment, a thin film dispersion method is adopted to prepare a high-capacity stepherin lipid composition, and the specific preparation method includes the following steps:

Weigh the prescription amount of lipid material and stepahin in Table 3 and add it to 1mL of chloroform, and dissolve it by ultrasonic; place it on a rotary evaporator, rotate for 45min at 45°C and 75rpm, and spread the film. If it is not hydrated immediately, it can be Place the film in a vacuum drying oven overnight; add 1mL of PBS with a pH value of 7.4, hydrate at 55°C, 100rpm for 45min; ultrasonicate in a cell disruptor for 2min, with an ultrasonic power of 45W and an interval of 2s, to obtain a colorless translucent high Lipid composition loaded with stepherin. The particle size of the liposome in the high-load stepherin lipid composition is 76nm, the Zeta potential is 4.0mV, and the drug loading is 8.48mg/mL (the stepherin accounts for the mass concentration of the fat-soluble drug lipid composition ), Stephalin accounted for 7.2% by mass of the total solid matter in the fat-soluble drug lipid composition, and the encapsulation efficiency was 99.73%.

Example 4

The ratio relationship of the raw materials of the fat-soluble drug lipid composition provided by this embodiment is shown in Table 4:

Table 4 The raw material mass ratio relationship of the fat-soluble drug lipid composition of this embodiment

In this example, the freeze-drying method is adopted to prepare a high-load stepherin lipid composition, and the specific preparation method includes the following steps:

Weigh the lipid material and stepherin in the prescription amount in Table 4 into a beaker, add 1mL of chloroform to ultrasonically dissolve to obtain solution a; weigh an appropriate amount of trehalose or mannose, and prepare a protective solution of a certain concentration with normal saline , to obtain solution b. Mix solution a and solution b, filter with a microporous membrane, pack in vials, pre-freeze in a -80°C refrigerator for 12 hours, put them in a freeze dryer for 24 hours and take them out to obtain a colorless translucent high Lipid composition loaded with stepherin. The liposome particle size in the prepared high-load stepherin lipid composition is 78nm, the Zeta potential is 1.9mV, and the drug loading is 9.82mg/mL (stepherin accounts for the total of the fat-soluble drug lipid composition). Mass concentration), Stephalin accounts for 6.0% by mass of the total solid matter in the fat-soluble drug lipid composition, and the encapsulation efficiency is 98.17%.

Example 5

The ratio relationship of the raw materials of the fat-soluble drug lipid composition provided by this embodiment is shown in Table 5:

Table 5 The raw material mass ratio relationship of the fat-soluble drug lipid composition of this embodiment

In this example, a high-pressure homogenization method is adopted to prepare a high-capacity stepahin lipid composition. The specific preparation method includes the following steps:

Weigh the prescription amount of lipid material and stephalin in Table 5, add it to 1mL chloroform, and dissolve it by ultrasonic; place it on a rotary evaporator, rotate at 45°C, 70rpm for 50min, and spread the film. If it is not hydrated immediately, it can be Place the film in a vacuum drying oven overnight; add 1mL of PBS with a pH value of 7.4, mix 5 times in a high-pressure homogenizer at 50,000kPa, and then mix 3 times at 100,000kPa, vacuum freeze-dry, and store at 4°C to obtain stephin with high drug loading lipid composition. The particle size of the liposome in the high-load stepherin lipid composition is 76nm, the Zeta potential is 2.9mV, and the drug loading is 16.99mg/mL (the stepherin accounts for the mass concentration of the fat-soluble drug lipid composition ), Stephalin accounted for 10.17% by mass of the total solid matter in the fat-soluble drug lipid composition, and the encapsulation efficiency was 99.92%.

Example 6

The ratio relationship of the raw materials of the fat-soluble drug lipid composition provided by this embodiment is shown in Table 6:

Table 6 The raw material mass ratio relationship of the fat-soluble drug lipid composition of this embodiment

In this embodiment, the ammonium sulfate gradient method is used to prepare a high-load stepahin lipid composition. The specific preparation method includes the following steps:

Weigh the lipid material in the prescription amount in Table 6 and dissolve it in chloroform, place it on a rotary evaporator, lay a film at 45°C, 70 rpm, and rotate for 45 minutes to obtain a lipid material film, and use 530mmol/L ammonium sulfate aqueous solution for lipid Hydrate the membrane of the plasma material, homogenize the obtained hydration solution under high pressure to prepare a blank lipid, dialyze with 160mmol/LNaCl at room temperature, 1h/time, a total of 3 times, add stepherin to incubate the loaded drug at 40°C , that is, to obtain a high drug-loaded stepherin lipid composition. The liposome particle size in the prepared high-load stepherin lipid composition is 96nm, the Zeta potential is 8.5mV, and the drug loading is 11.00mg/mL (stepherin accounts for 1% of the fat-soluble drug lipid composition). Mass concentration), Stephalin accounted for 9.0% by mass of the total solid matter in the fat-soluble drug lipid composition, and the encapsulation efficiency was 99.97%.

Example 7

The ratio relationship of the raw materials of the fat-soluble drug lipid composition provided by this embodiment is shown in Table 7:

Table 7 The raw material mass ratio relationship of the fat-soluble drug lipid composition of this embodiment

In this embodiment, a thin film dispersion method is adopted to prepare a high-capacity stepherin lipid composition, and the specific preparation method includes the following steps:

Weigh DOPC, DSPE-MPEG2000 and ivermectin in the prescription amount in Table 7 respectively, add chloroform, ultrasonically dissolve, place on a rotary evaporator at 45°C, 75rpm, and rotate for more than 45 minutes to lay a film; add 2mL of pH value of 7.4 PBS, 55°C, 100 rpm, hydrated for 45 minutes, and then ultrasonically dispersed. The ivermectin lipid composition with high drug load is obtained. Ivermectin drug loading is 15.00mg/mL (ivermectin accounts for the mass concentration of described fat-soluble drug lipid composition), and ivermectin accounts for total solid matter in described fat-soluble drug lipid composition The mass percent content is 12.93%, and the encapsulation efficiency is 100%.

Comparative example 1

The raw material mass ratio relationship of the fat-soluble drug lipid composition provided in this comparative example is shown in Table 8:

Table 8 The raw material mass ratio relationship of the fat-soluble drug lipid composition of this comparative example

This comparative example adopts thin-film dispersion method to prepare stepherin lipid composition, and concrete preparation method comprises the following steps:

Weigh the HSPC, DSPG, cholesterol and stephalin in the prescription amount in Table 8 respectively, add chloroform, ultrasonically dissolve, place on a rotary evaporator at 45°C, 75rpm, and rotate for more than 45 minutes to spread the film; add 1mL of PBS with a pH value of 7.4 , 55°C, 100rpm, hydration for 45 minutes, and then ultrasonic dispersion.

The experimental results showed that after the film was laid, vacuuming revealed that there was a white gelatinous solid at the bottom of the eggplant-shaped bottle, and a thin film was formed around the eggplant-shaped bottle. After hydration, it is a white milky opaque liquid with insoluble matter at the bottom. After ultrasound, the color becomes slightly transparent, but it is still an opaque milky white liquid.

Comparative example 2

The raw material mass ratio relationship of the fat-soluble drug lipid composition provided in this comparative example is as shown in Table 9:

Table 9 The raw material quality ratio relationship of the fat-soluble drug lipid composition of this comparative example

This comparative example adopts thin-film dispersion method to prepare stepherin lipid composition, and concrete preparation method comprises the following steps:

Weigh the HSPC, DSPG, cholesterol and stephalin in the prescription amount in Table 9 respectively, add chloroform, ultrasonically dissolve, place on a rotary evaporator at 45°C, 75rpm, and rotate for more than 45 minutes to spread the film; add 2mL of PBS with a pH of 7.4, 55°C, 100rpm, hydration for 45 minutes, and then ultrasonic dispersion.

The experimental results show that the film laying state is normal, the hydration state is good, the fluidity of the composition is good after ultrasound, and the color is slightly lighter than that of Comparative Example 1, but it is still an opaque liquid, which is different from the normal translucent liquid with light blue opalescence Inconsistent state.

Comparative example 3

The raw material mass ratio relationship of the fat-soluble drug lipid composition provided in this comparative example is shown in Table 8:

Table 10 Comparative Example 3 The raw material mass ratio relationship of the fat-soluble drug lipid composition

This comparative example adopts thin-film dispersion method to prepare stepherin lipid composition, and concrete preparation method comprises the following steps:

Weigh the HSPC, DSPG, cholesterol and stephalin in the prescription amounts in Table 10, add chloroform, dissolve by ultrasonication, place on a rotary evaporator at 45°C, 75rpm, and rotate for more than 45 minutes to lay a film; add 2mL of PBS with a pH of 7.4, 55 °C, 100 rpm, hydration for 45 minutes, and then ultrasonic dispersion.

In the preparation process, it was found that DPPE was difficult to dissolve in chloroform, and it was insoluble by ultrasound, so it was difficult to form a film. However, the subsequent hydration state is good and the final composition has good fluidity.

Table 11 shows the results of the lipid compositions with high loads of stepherin or ivermectin prepared in various examples and comparative examples of the present invention.

The basic properties of table 11 high drug loading lipid composition

It can be concluded from Table 11 that, compared with Comparative Examples 1-3, the amount of drug-loaded liposomes in the high-load stepherin or ivermectin lipid composition prepared in Examples 1-7 of the present invention is smaller Small size, high drug loading capacity and high encapsulation efficiency. The main reason for the poor experimental effect in Comparative Examples 1-3 may be that the composition of the prescription and its dosage have poor tolerance/compatibility to the drug in the invention, resulting in its inability to form a uniform and stable drug with a higher amount of drug. lipid solution.

Although the foregoing embodiment has described the present invention in detail, it is only a part of the embodiments of the present invention, rather than all embodiments, and other embodiments can also be obtained according to the present embodiment without inventive step, and these embodiments are all Belong to the protection scope of the present invention.

Claims (10)

1. a fat-soluble drug lipid composition, is characterized in that, comprises drug-loaded liposome and dispersion solvent; Described drug-loaded liposome comprises fat-soluble medicine and the lipid material that entraps described fat-soluble medicine, The fat-soluble drug includes one or more of stepherin, stepherin salt, stepherin derivative, ivermectin, ivermectin salt and ivermectin derivative, and the lipid The material includes one or more of phosphatidylcholine compounds, phosphatidylethanolamine compounds, phosphatidylglycerol compounds, acylphosphatidic acid compounds, phosphatidylserine compounds, and lysophospholipid compounds; the lipid material The mass concentration in the dispersion solvent is 10-300 mg/mL; In the fat-soluble drug lipid composition, the drug loading amount of the fat-soluble drug is more than or equal to 8 mg/mL. 2. fat-soluble drug lipid composition according to claim 1, is characterized in that, described lipid material comprises hydrogenated soybean lecithin, egg yolk lecithin, dioleoyl lecithin, dipalmitoyl lecithin, dihard One or more of fatty acylphosphatidylcholine, 2-palmitoylphosphatidylglycerol, distearoylphosphatidylethanolamine-polyethylene glycol. 3. fat-soluble drug lipid composition according to claim 2, is characterized in that, described lipid material comprises egg yolk lecithin, dioleoyl lecithin, dipalmitoyl lecithin, distearoyl phosphatidyl choline One or more of alkali, 2-palmitoylphosphatidylglycerol, distearoylphosphatidylethanolamine-polyethylene glycol. 4. The fat-soluble drug lipid composition according to claim 1, characterized in that, the drug loading amount of the fat-soluble drug is 8-20 mg/mL. 5. The fat-soluble drug lipid composition according to claim 1, wherein the dispersion solvent comprises one or more of water, physiological saline, glucose solution or phosphate buffered saline. 6. fat-soluble drug lipid composition according to claim 1, is characterized in that, described fat-soluble drug lipid composition also comprises cholesterol; The mass ratio of described cholesterol and described fat-soluble drug is (0～ 1): (0.5～2). 7. The preparation method of the fat-soluble drug lipid composition described in any one of claims 1 to 6, comprising film dispersion method, reverse evaporation method, solvent injection method, freeze-drying method, French pressure method, double emulsion method, high pressure Any one of homogeneous method, pH gradient method, microfluidic method and microfluidic method. 8. The application of the fat-soluble drug lipid composition described in any one of claims 1 to 6 or the fat-soluble drug lipid composition prepared by the preparation method described in claim 7 in the preparation of medicines, characterized in that , the drugs include drugs for treating leukopenia, drugs for treating silicosis, drugs for treating pneumoconiosis, drugs for treating snake bites, drugs for treating alopecia areata, drugs for treating alopecia, drugs for treating essential thrombocytopenic purpura, Medicines for mouth ulcers, medicines for endotoxic shock, antimalarial medicines, immunomodulatory medicines, antiviral medicines, or antineoplastic medicines. 9. The application according to claim 8, characterized in that the dosage form of the drug comprises oral preparations, external preparations or injection preparations. 10. The application according to claim 9, characterized in that, the external preparations include transdermal preparations, ophthalmic preparations or nasal cavity preparations.

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