CN110522726B - Preparation method of dipheny hydrochloride liposome and dipheny hydrochloride liposome - Google Patents

Preparation method of dipheny hydrochloride liposome and dipheny hydrochloride liposome Download PDF

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CN110522726B
CN110522726B CN201910880245.0A CN201910880245A CN110522726B CN 110522726 B CN110522726 B CN 110522726B CN 201910880245 A CN201910880245 A CN 201910880245A CN 110522726 B CN110522726 B CN 110522726B
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韩翠艳
潘涛
肖雪
费培哲
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Abstract

The invention provides a preparation method of a diphenoxylate hydrochloride liposome and the diphenoxylate hydrochloride liposome, belonging to the technical field of medicines, and the method comprises the following steps: 1) mixing the diphenhydrasol hydrochloride, the lecithin, the cholesterol and the absolute ethyl alcohol to obtain an organic phase solution; 2) injecting the organic phase solution in the step 1) into water under stirring to obtain liposome suspension; 3) removing ethanol in the liposome suspension, fixing the volume, preserving the heat in a water bath, and performing ultrasonic treatment to obtain a diphenhydrase hydrochloride liposome; the mass ratio of the lecithin to the cholesterol is (5.5-6.5): 1; the mass ratio of the lecithin to the diphenhydrasol hydrochloride is (41-43): 1. the dipheny xol hydrochloride liposome provided by the invention has the advantages of high encapsulation efficiency, good reproducibility, large drug-loading rate and slow-release effect.

Description

Preparation method of dipheny hydrochloride liposome and dipheny hydrochloride liposome
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a preparation method of a diphenoxylate hydrochloride liposome and the diphenoxylate hydrochloride liposome.
Background
The dipheny hcl is mainly used for parkinsonism, paralysis agitans or paralysis agitans caused by arteriosclerosis in clinic, and the currently marketed dipheny hcl pharmaceutical dosage form is mainly tablets, and has the defects of slow effect taking, low absorption efficiency, and a series of adverse reactions such as gastrointestinal tracts and the like caused by long-term use.
The liposome is a multi-layer microcapsule consisting of orderly arranged lipid bilayers, and the size of the liposome is in the range of dozens of nanometers to dozens of micrometers; the nasal administration has no gastrointestinal degradation effect, no liver first pass effect, and quick action after administration. At present, liposomes have attracted extensive attention in various research fields at home and abroad as good encapsulating carriers, and especially, in the research and design process of innovative dosage forms, scientists often select liposomes as the encapsulating carriers. Nasal administration is also widely studied because of its lack of gastrointestinal degradation, no liver first pass effect, rapid drug absorption, and rapid onset of action after administration; the nasal administration preparation using liposome as carrier is the hot spot of research in recent years. The liposome has the advantages that the sustained release effect on the drug is realized through the phospholipid bilayer of the liposome, the liposome has the effects of stabilizing the blood concentration and protecting the drug from being decomposed by enzyme in the absorption and transportation processes, and the stimulation and toxicity of the drug on the nasal mucosa can be effectively avoided and reduced, so that the curative effect of the drug is improved. As Parkinson patients are often seen in the elderly, the liposome nasal mucosa is more convenient to permeate for administration compared with the tablet sold in the market.
Therefore, the diphenhydrasol hydrochloride is prepared into liposome to be administrated through nasal cavity, so that the defects of oral tablets can be overcome. However, the research on preparing the diphenoxylate hydrochloride into the liposome is less, and the effects of entrapment rate, drug loading capacity, release degree and the like are poorer.
Disclosure of Invention
In view of the above, the present invention aims to provide a method for preparing a diphenoxylate hydrochloride liposome and a diphenoxylate hydrochloride liposome; the diphenhydramine hydrochloride liposome prepared by the method has high entrapment rate, large drug-loading rate and slow-release effect.
In order to achieve the above purpose, the invention provides the following technical scheme:
the invention provides a preparation method of a diphenoxylate hydrochloride liposome, which comprises the following steps:
1) mixing the diphenhydrasol hydrochloride, the lecithin, the cholesterol and the absolute ethyl alcohol to obtain an organic phase solution;
2) injecting the organic phase solution in the step 1) into water under stirring to obtain liposome suspension;
3) removing ethanol in the liposome suspension, fixing the volume, preserving the heat in a water bath, and performing ultrasonic treatment to obtain a diphenhydrase hydrochloride liposome;
the mass ratio of the lecithin to the cholesterol is (5.5-6.5): 1; the mass ratio of the lecithin to the diphenhydrasol hydrochloride is (41-43): 1;
the temperature of the water in the step 2) is 48-52 ℃; and 3) keeping the temperature of the water bath at 48-52 ℃ in the step 3), and keeping the temperature of the water bath for 50-70 min.
Preferably, the mass-to-volume ratio of the trihexyphenidyl hydrochloride to the absolute ethyl alcohol in the step 1) is 1mg (4-6) mL.
Preferably, the volume ratio of the absolute ethyl alcohol in the step 1) to the water in the step 2) is (0.9-1.1): (0.9-1.1).
Preferably, the rotation speed of the stirring in the step 2) is 250-350 rpm.
Preferably, after all the organic phase solution in the step 2) is injected into water, stirring is continued for 25-35 min, and a liposome suspension is obtained.
Preferably, the injection speed of the organic phase solution in the step 2) is (8-12) mL/h.
Preferably, the mass ratio of the lecithin to the cholesterol is 6.02: 1; the mass ratio of the lecithin to the diphenhydrasol hydrochloride is 41.96: 1.
Preferably, the power of the ultrasonic treatment in the step 3) is 180-220W; the ultrasonic treatment time is 8-12 min.
Preferably, the method for removing the absolute ethanol in the step 3) is a rotary evaporation method.
The invention provides the diphenoxylate hydrochloride liposome prepared by the preparation method, and the entrapment rate of the diphenoxylate hydrochloride liposome is more than 89%; the drug loading rate is more than 1.65 percent, and the 12h in vitro release rate is more than 84 percent.
The invention has the beneficial effects that: the invention provides a preparation method of a trihexyphenidyl hydrochloride liposome, which utilizes an ethanol injection method, and controls the mass ratio of lecithin to cholesterol, the mass ratio of lecithin to trihexyphenidyl hydrochloride, the ratio of organic phase solution to water, the temperature and time of water bath heat preservation and other parameters to obtain the trihexyphenidyl hydrochloride liposome with high encapsulation efficiency, good reproducibility and large drug-loading capacity.
Drawings
FIG. 1 is an electron micrograph of TH liposomes (x 260000) from example 2;
figure 2 is a time-release profile of the TH liposomes and drug substance of example 2.
Detailed Description
The invention provides a preparation method of a diphenoxylate hydrochloride liposome, which comprises the following steps: 1) mixing the diphenhydrasol hydrochloride, the lecithin, the cholesterol and the absolute ethyl alcohol to obtain an organic phase solution; 2) injecting the organic phase solution in the step 1) into water under stirring to obtain liposome suspension; 3) removing ethanol in the liposome suspension, fixing the volume, preserving the heat in a water bath, and performing ultrasonic treatment to obtain a diphenhydrase hydrochloride liposome; the mass ratio of the lecithin to the cholesterol is (5.5-6.5): 1; the mass ratio of the lecithin to the diphenhydrasol hydrochloride is (41-43): 1; the temperature of the water in the step 2) is 48-52 ℃; and 3) keeping the temperature of the water bath at 48-52 ℃ in the step 3), and keeping the temperature of the water bath for 50-70 min.
In the present invention, diphenhydrasol hydrochloride, lecithin, cholesterol and absolute ethanol are mixed to obtain an organic phase solution. In the invention, the mass ratio of the lecithin to the cholesterol is (5.5-6.5): 1, preferably (5.8-6.2): 1, more preferably 6.02: 1; the mass ratio of the lecithin to the diphenhydrasol hydrochloride is (41-43): 1, preferably (41.5-42.5): 1, more preferably 41.96: 1. In the invention, the mass-to-volume ratio of the trihexyphenidyl hydrochloride to the absolute ethyl alcohol is preferably 1mg (4-6) mL, more preferably 1mg (4.5-5.5) mL, and most preferably 1mg:5 mL. The sources of the diphenhydrasol hydrochloride, the lecithin, the cholesterol and the absolute ethyl alcohol are not particularly limited, and the conventional commercial products in the field can be adopted. In the invention, the anhydrous ethanol is used for fully dissolving the diphenhydramine hydrochloride, the lecithin and the cholesterol to obtain an organic phase solution.
After the organic phase solution is obtained, the organic phase solution is injected into water under the condition of stirring to obtain liposome suspension. In the present invention, the rotation speed of the stirring is preferably 250 to 350rpm, more preferably 280 to 320rpm, and most preferably 300 rpm. In the invention, the injection speed of the organic phase solution is preferably (8-12) mL/h, more preferably (9-11) mL/h, and most preferably 10 mL/h. In the present invention, the injection is preferably realized by using a syringe, and the injection can be selected from mechanical injection and manual injection. According to the invention, preferably, after all the organic phase solution is injected into water, the mixture is continuously stirred for 25-35 min to obtain liposome suspension; the stirring time is preferably 28-32 min, and more preferably 30 min; the rotation speed of the stirring is consistent with the limitation of the rotation speed of the stirring, and the details are not repeated herein. In the invention, the temperature of the water is preferably 48-52 ℃, more preferably 49-51 ℃, and most preferably 50.33 ℃. In the present invention, the water is preferably purified water.
After the liposome suspension is obtained, ethanol in the liposome suspension is removed, and the diphenyol hydrochloride liposome is obtained after constant volume, water bath heat preservation and ultrasonic treatment. In the present invention, the method for removing anhydrous ethanol is preferably a rotary evaporation method; the temperature and time for removing the absolute ethyl alcohol by rotary evaporation are not particularly limited, and the absolute ethyl alcohol can be completely removed at normal temperature. In the invention, the constant volume is preferably carried out by using water, and the volume of the feed liquid after constant volume is consistent with that of the liposome suspension. In the present invention, the water is preferably purified water; the temperature of the water bath heat preservation is preferably 48-52 ℃, more preferably 49-51 ℃, and most preferably 50 ℃, and the time of the water bath heat preservation is preferably 50-70 min, more preferably 55-65 min, and most preferably 60 min. After the water bath is insulated, ultrasonic treatment is carried out, wherein the ultrasonic treatment is preferably ultrasonic treatment of a probe, and the power of the ultrasonic treatment is preferably 180-220W, more preferably 200W; the time of ultrasonic treatment is preferably 8-12 min, and more preferably 10 min; the ultrasonication acts to control the particle size uniformity of the liposomes.
The invention also provides the diphenoxylate hydrochloride liposome prepared by the preparation method, and the entrapment rate of the diphenoxylate hydrochloride liposome is more than 89%; the drug loading rate is more than 1.65 percent, and the 12h in vitro release rate is more than 84 percent. In a specific implementation process of the invention, the diphenoxylate hydrochloride liposome has uniform size, round shape, uniform distribution, good liposome shape, the entrapment rate of 91.5%, the particle size of 72.2nm, the zeta potential of 6.3mv, the drug-loading rate of 1.67% and the in-vitro release degree of 81.4% within 12h, and the release conforms to first-order release and shows a certain slow release effect.
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
The apparatus and reagents used in the following examples and comparative examples are summarized below:
instrument, apparatus
Figure BDA0002205660940000051
The assay methods referred to in the examples are summarized below:
formulation Properties study morphology: transmission electron microscopy scans the liposome morphology. Diluting TH liposome with double distilled water, negatively dyeing with 2% phosphotungstic acid solution, dripping onto the power surface of copper mesh, drying, and scanning with transmission electron microscope.
Particle diameter and zeta potential: the particle size and the distribution of the liposome are measured under a ZEN 3690-nanometer particle size analyzer. Diluting 1mL of the liposome by 10 times, placing a proper amount of the liposome into a cuvette, placing the cuvette into a sample cell for determination, and repeating the determination for 3 times to obtain the particle size and potential distribution of the TH liposome.
Encapsulation efficiency and drug loading:
encapsulation efficiency: precisely measuring drug-containing liposome 0.1mL, precisely adding acetonitrile 0.16mL, demulsifying, precisely adding 0.24mL 0.1% triethylamine (pH adjusted to 4.0 with phosphoric acid), shaking, filtering with 0.22 μm microporous membrane, and sampling according to the following chromatographic conditions to obtain total concentration CGeneral assembly(ii) a And taking a proper amount of TH liposome to an ultrafiltration centrifugal tube, and centrifuging for 30min at 5 ℃ under the condition of 10000 r/min. The supernatant was taken at 0.1mL and 0.16mL acetonitrile and 0.24mL0 were added.1% triethylamine (pH 4.0 adjusted by phosphoric acid), shaking, filtering with 0.22 μm microporous membrane, injecting sample under the following chromatographic conditions, and measuring the concentration of free drug CFree form(ii) a The encapsulation efficiency was calculated according to the following formula.
Figure BDA0002205660940000061
In the formula CGeneral assembly: total dose encapsulated versus unencapsulated in the system; cFree form: unencapsulated drug quantity in a system
Chromatographic condition sample introduction conditions: octadecylsilane chemically bonded silica is used as a filling agent; taking 0.1% triethylamine solution (pH value is adjusted to 4.0 by phosphoric acid) to acetonitrile (60/40) as a mobile phase, and the sample amount is 20 mu l; the sample introduction speed is 1.0ml/min, and the detection wavelength is as follows: 210 nm.
The drug-loading rate measuring method is that the adding amount of each component in the prescription is calculated according to the following formula.
Figure BDA0002205660940000062
In the formula MFat and oil: the amount of the liposome traditional Chinese medicine; mCarrier: total amount of carrier
Determination of the degree of in vitro Release: precisely sucking 10mL of TH liposome, adding into a pretreated dialysis bag, fastening the bag opening, placing in a beaker filled with 50mLPBS medium, regularly sucking 500uL of dialysate in a constant-temperature water bath at (37 +/-1) DEG C at 100r/min, supplementing with an equal amount of constant-temperature PBS medium, precisely sucking 10mLTH liposome, placing in a 50mL volumetric flask, demulsifying with acetonitrile, and diluting to a certain volume with PBS to obtain the total dosage. And meanwhile, the release condition of the TH bulk drug is considered.
Example 1
100mg of lecithin, 10mg of cholesterol and 2mg of diphenyol hydrochloride are precisely weighed and fully dissolved by 10mL of absolute ethyl alcohol to form an organic phase. Weighing 10mL of purified water in a beaker, slowly injecting the oil phase into the purified water by using an injector under the magnetic stirring at the film forming temperature of 50 ℃ and the rotating speed of 300rpm, and continuously stirring for 30min to obtain the trihexyphenidyl hydrochloride liposome suspension. And (4) carrying out rotary evaporation at room temperature to remove the absolute ethyl alcohol. And finally, adding purified water to a constant volume of 10mL, preserving the heat in a water bath at 50 ℃ for 1h, cooling to 25 ℃, and performing ultrasonic treatment on the probe for 10min to obtain the diphenhydrasol hydrochloride liposome.
Comparative example 1
Accurately weighing 100mg of lecithin and 10mg of cholesterol and TH 2mg into a 100mL round bottom rotary evaporation bottle, adding 10mL of absolute ethyl alcohol to dissolve the lecithin and the cholesterol, placing the solution on a rotary evaporator, carrying out rotary evaporation for 40min under the condition of 50 ℃ water bath, removing an organic solvent until a uniform lipid film is formed on the wall of the bottle, taking 10mL of purified water by using a measuring cylinder, placing the solution in the round bottom flask, and hydrating for 40min under the condition of 50 ℃ water bath. The probe is used for ultrasonic treatment for 10 min. And (4) carrying out constant volume treatment on the liposome after ultrasonic treatment to 10mL to obtain the TH liposome.
Comparative example 2
Precisely weighing 100mg of lecithin and 10mg of cholesterol, dissolving in 20mL of absolute ethanol, and taking 0.12 mol.L-160mL of the aqueous solution of ammonium sulfate was preheated to 50 ℃ and the above ethanol solution was slowly poured into the ammonium sulfate solution under stirring. Stirring in water bath at 50 deg.C for 2 hr, volatilizing ethanol, and performing ultrasonic treatment for 10min to obtain blank liposome suspension. The prepared blank liposome suspension is filled in a dialysis bag, the two ends of the bag are sealed tightly, and the bag is placed in a beaker filled with 500mL of physiological saline and dialyzed for 3 times at room temperature, and each time lasts for 2 hours. Adding TH water solution (100mg → 6ml, stirring and dissolving at 80 deg.C) into the dialyzed liposome, stirring, performing ultrasonic treatment with probe for 10min, and keeping temperature in 37 deg.C water bath for 30min to obtain TH liposome.
The encapsulation efficiency, particle size and zeta potential of the liposomes of diphenoxylate obtained by preparation in example 1, comparative example 1 and comparative example 2 were measured, and the results are shown in table 1.
TABLE 1 measurement results of three indexes of liposome prepared by three methods
Figure BDA0002205660940000071
From the results in Table 1, it can be seen that: among the three different preparation methods, the liposome prepared by the ethanol injection method has the highest encapsulation efficiency, the thin film dispersion method is inferior, and the ammonium sulfate gradient method is the lowest; the liposome prepared by the ethanol injection method has the smallest particle size, the film dispersion method is inferior, and the ammonium sulfate gradient method is the largest; the zeta potential absolute value of the liposome prepared by the film dispersion method is highest, the ethanol injection method is second, the ammonium sulfate gradient method is lowest, and the ethanol injection method is worse than the film dispersion method in potential evaluation, but the other two indexes are optimal.
Example 2
83.92mg of lecithin, 13.94mg of cholesterol and 2mg of diphenhydrasol hydrochloride were precisely weighed and sufficiently dissolved in 10mL of absolute ethanol to form an organic phase. Weighing 10mL of purified water in a beaker, slowly injecting the oil phase into the purified water by using an injector under the magnetic stirring at the film forming temperature of 50 ℃ and the rotating speed of 300rpm, and continuously stirring for 30min to obtain the trihexyphenidyl hydrochloride liposome suspension. And (4) carrying out rotary evaporation at room temperature to remove the absolute ethyl alcohol. And finally, adding purified water to a constant volume of 10mL, preserving the heat in a water bath at 50 ℃ for 1h, cooling to 25 ℃, and performing ultrasonic treatment on the probe for 10min to obtain the diphenhydrasol hydrochloride liposome.
For morphological examination, the picture of the diphenoxylate hydrochloride liposome taken by a transmission electron microscope is shown in FIG. 1. It can be seen from the photographs taken by transmission electron microscopy at a magnification of twenty-sixty thousand times: the liposome has uniform size, round shape, uniform distribution and good liposome shape.
Particle size and zeta potential investigation: the particle size and zeta potential of the three groups of liposomes were measured separately by a nanometer particle sizer and the results are shown in table 2. From the results of table 2, it can be seen that: calculating the average value of the particle size to be 73.2 nm; the zeta potential averaged 6.3 mv.
TABLE 2 measurement results of particle diameter and zeta potential
Figure BDA0002205660940000081
The encapsulation efficiency and drug loading were examined and the results are shown in Table 3. From the results of table 3, it can be seen that: the average liposome entrapment rate was 91.5% and the drug loading was 1.69%.
TABLE 3 encapsulation efficiency and drug loading measurements
Figure BDA0002205660940000082
Figure BDA0002205660940000091
In vitro release degree investigation: the release time of the TH liposome and the bulk drug thereof in the release medium versus the accumulated release degree is plotted into a curve as shown in figure 2, and the in vitro release data of the TH liposome is processed by a single exponential function and a Higuchi equation respectively to obtain a regression equation as shown in table 4. As can be seen from fig. 2: within 12h, the TH liposome has lower in-vitro accumulated release degree than the bulk drug. The release of the bulk drug is 95.9% at 240min, the release tends to be complete, the liposome releases 64.4% at the time, and 84.1% at 12h, which indicates that the liposome has more sustained release effect.
TABLE 4 Liposome different Release model fitting equation
Figure BDA0002205660940000092
Example 3
84mg of lecithin, 14mg of cholesterol and 2mg of diphenyol hydrochloride are precisely weighed and fully dissolved by 10mL of absolute ethyl alcohol to form an organic phase. Weighing 10mL of purified water in a beaker, slowly injecting the oil phase into the purified water by using an injector under the magnetic stirring at the film forming temperature of 50 ℃ and the rotating speed of 300rpm, and continuously stirring for 30min to obtain the trihexyphenidyl hydrochloride liposome suspension. And (4) carrying out rotary evaporation at room temperature to remove the absolute ethyl alcohol. And finally, adding purified water to a constant volume of 10mL, preserving the heat in a water bath at 50 ℃ for 1h, cooling to 25 ℃, and performing ultrasonic treatment on the probe for 10min to obtain the diphenhydrasol hydrochloride liposome. The encapsulation efficiency was determined to be 61.2%.
Example 4
82mg of lecithin, 20.5mg of cholesterol and 2mg of diphenhydrasol hydrochloride are precisely weighed and fully dissolved by 10mL of absolute ethyl alcohol to form an organic phase. Weighing 10mL of purified water in a beaker, slowly injecting the oil phase into the purified water by using an injector under the magnetic stirring at the film forming temperature of 50 ℃ and the rotating speed of 300rpm, and continuously stirring for 30min to obtain the trihexyphenidyl hydrochloride liposome suspension. And (4) carrying out rotary evaporation at room temperature to remove the absolute ethyl alcohol. And finally, adding purified water to a constant volume of 10mL, preserving the heat in a water bath at 50 ℃ for 1h, cooling to 25 ℃, and performing ultrasonic treatment on the probe for 10min to obtain the diphenhydrasol hydrochloride liposome. The encapsulation efficiency was determined to be 24.3%.
Example 5
82mg of lecithin, 14.9mg of cholesterol and 2mg of diphenhydrasol hydrochloride were precisely weighed and sufficiently dissolved in 10mL of absolute ethanol to form an organic phase. Weighing 10mL of purified water in a beaker, slowly injecting the oil phase into the purified water by using an injector under the magnetic stirring at the film forming temperature of 50 ℃ and the rotating speed of 300rpm, and continuously stirring for 30min to obtain the trihexyphenidyl hydrochloride liposome suspension. And (4) carrying out rotary evaporation at room temperature to remove the absolute ethyl alcohol. And finally, adding purified water to a constant volume of 10mL, preserving the heat in a water bath at 50 ℃ for 1h, cooling to 25 ℃, and performing ultrasonic treatment on the probe for 10min to obtain the diphenhydrasol hydrochloride liposome.
Comparative example 3
40mg of lecithin, 10mg of cholesterol and 2mg of diphenyol hydrochloride are precisely weighed and fully dissolved by 10mL of absolute ethyl alcohol to form an organic phase. Weighing 10mL of purified water in a beaker, slowly injecting the oil phase into the purified water by using an injector under the magnetic stirring at the film forming temperature of 50 ℃ and the rotating speed of 300rpm, and continuously stirring for 30min to obtain the trihexyphenidyl hydrochloride liposome suspension. And (4) carrying out rotary evaporation at room temperature to remove the absolute ethyl alcohol. And finally, adding purified water to a constant volume of 10mL, preserving the heat in a water bath at 50 ℃ for 1h, cooling to 25 ℃, and performing ultrasonic treatment on the probe for 10min to obtain the diphenhydrasol hydrochloride liposome. The encapsulation efficiency was determined to be 23.1%.
Comparative example 4
120mg of lecithin, 10mg of cholesterol and 2mg of diphenyol hydrochloride are precisely weighed and fully dissolved by 10mL of absolute ethyl alcohol to form an organic phase. Weighing 10mL of purified water in a beaker, slowly injecting the oil phase into the purified water by using an injector under the magnetic stirring at the film forming temperature of 50 ℃ and the rotating speed of 300rpm, and continuously stirring for 30min to obtain the trihexyphenidyl hydrochloride liposome suspension. And (4) carrying out rotary evaporation at room temperature to remove the absolute ethyl alcohol. And finally, adding purified water to a constant volume of 10mL, preserving the heat in a water bath at 50 ℃ for 1h, cooling to 25 ℃, and performing ultrasonic treatment on the probe for 10min to obtain the diphenhydrasol hydrochloride liposome. The encapsulation efficiency was determined to be 24.7%.
Comparative example 5
200mg of lecithin, 10mg of cholesterol and 2mg of diphenyol hydrochloride are precisely weighed and fully dissolved by 10mL of absolute ethyl alcohol to form an organic phase. Weighing 10mL of purified water in a beaker, slowly injecting the oil phase into the purified water by using an injector under the magnetic stirring at the film forming temperature of 50 ℃ and the rotating speed of 300rpm, and continuously stirring for 30min to obtain the trihexyphenidyl hydrochloride liposome suspension. And (4) carrying out rotary evaporation at room temperature to remove the absolute ethyl alcohol. And finally, adding purified water to a constant volume of 10mL, preserving the heat in a water bath at 50 ℃ for 1h, cooling to 25 ℃, and performing ultrasonic treatment on the probe for 10min to obtain the diphenhydrasol hydrochloride liposome. The encapsulation efficiency was determined to be 17.0%.
Comparative example 6
120mg of lecithin, 10mg of cholesterol and 2mg of diphenyol hydrochloride are precisely weighed and fully dissolved by 10mL of absolute ethyl alcohol to form an organic phase. Weighing 30mL of purified water in a beaker, slowly injecting the oil phase into the purified water by using an injector under the magnetic stirring at the film forming temperature of 50 ℃ and the rotating speed of 300rpm, and continuously stirring for 30min to obtain the trihexyphenidyl hydrochloride liposome suspension. And (4) carrying out rotary evaporation at room temperature to remove the absolute ethyl alcohol. And finally, adding purified water to a constant volume of 10mL, preserving the heat in a water bath at 50 ℃ for 1h, cooling to 25 ℃, and performing ultrasonic treatment on the probe for 10min to obtain the diphenhydrasol hydrochloride liposome. The encapsulation efficiency was determined to be 64.6%.
Comparative example 7
120mg of lecithin, 10mg of cholesterol and 2mg of diphenyol hydrochloride are precisely weighed and fully dissolved by 10mL of absolute ethyl alcohol to form an organic phase. Weighing 20mL of purified water into a beaker, slowly injecting the oil phase into the purified water by using an injector under the magnetic stirring at the film forming temperature of 50 ℃ and the rotating speed of 300rpm, and continuously stirring for 30min to obtain the trihexyphenidyl hydrochloride liposome suspension. And (4) carrying out rotary evaporation at room temperature to remove the absolute ethyl alcohol. And finally, adding purified water to a constant volume of 10mL, preserving the heat in a water bath at 50 ℃ for 1h, cooling to 25 ℃, and performing ultrasonic treatment on the probe for 10min to obtain the diphenhydrasol hydrochloride liposome. The encapsulation efficiency was determined to be 70.9%.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.

Claims (10)

1. A preparation method of a diphenhydrasol hydrochloride liposome is characterized by comprising the following steps:
1) mixing the diphenhydrasol hydrochloride, the lecithin, the cholesterol and the absolute ethyl alcohol to obtain an organic phase solution;
2) injecting the organic phase solution in the step 1) into water under stirring to obtain liposome suspension;
3) removing ethanol in the liposome suspension, fixing the volume, preserving the heat in a water bath, and performing ultrasonic treatment to obtain a diphenhydrase hydrochloride liposome;
the mass ratio of the lecithin to the cholesterol is (5.5-6.5): 1; the mass ratio of the lecithin to the diphenhydrasol hydrochloride is (41-43): 1;
the temperature of the water in the step 2) is 48-52 ℃; and 3) keeping the temperature of the water bath at 48-52 ℃ in the step 3), and keeping the temperature of the water bath for 50-70 min.
2. The preparation method of the diphenhydrase hydrochloride liposome according to claim 1, wherein the mass-to-volume ratio of the diphenhydrase hydrochloride to the absolute ethyl alcohol in the step 1) is 1mg (4-6) mL.
3. The method for preparing the diphenoxylate hydrochloride liposome according to claim 1, wherein the volume ratio of the absolute ethanol in the step 1) to the water in the step 2) is (0.9-1.1): (0.9-1.1).
4. The method for preparing the diphenyol hydrochloride liposome according to claim 1 or 3, wherein the stirring speed in the step 2) is 250-350 rpm.
5. The method for preparing the diphenyol hydrochloride liposome according to claim 4, wherein the liposome suspension is obtained by continuously stirring for 25-35 min after the organic phase solution is completely injected into water in the step 2).
6. The method for preparing the diphenoxylate hydrochloride liposome according to claim 1, wherein the injection speed of the organic phase solution in the step 2) is (8-12) mL/h.
7. The method for preparing the diphenoxylate hydrochloride liposome according to claim 1, wherein the mass ratio of the lecithin to the cholesterol is 6.02: 1; the mass ratio of the lecithin to the diphenhydrasol hydrochloride is 41.96: 1.
8. The method for preparing the diphenyol hydrochloride liposome according to claim 1, wherein the power of the ultrasonic treatment in the step 3) is 180-220W; the ultrasonic treatment time is 8-12 min.
9. The method for preparing the diphenoxylate hydrochloride liposome according to claim 1, wherein the method for removing the absolute ethanol in the step 3) is a rotary evaporation method.
10. The diphenhydrase hydrochloride liposome prepared by the preparation method of any one of claims 1-9, wherein the entrapment rate of the diphenhydrase hydrochloride liposome is more than 89%; the drug loading rate is more than 1.65 percent, and the 12h in vitro release rate is more than 84 percent.
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