CN112137968A - Formula and process for preparing liposome freeze-dried powder - Google Patents
Formula and process for preparing liposome freeze-dried powder Download PDFInfo
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Abstract
The invention provides a preparation formula of liposome freeze-dried powder, which comprises the following components: liposomes, excipients, water; the specific formula is prepared from the following raw materials in percentage by weight: 50% of liposome, 5-15% of excipient and the balance of water; the composition of the liposome is as follows: phospholipid 1-20%, cholesterol 0.1-10%, Tween 0.2-2%, trehalose 5-10%, active substance 0.2-20%, anhydrous ethanol 2-30% and water in balance. The invention has the following advantages: the liposome of the invention greatly improves the flexibility of the liposome due to the existence of the Tween, and can lead the liposome to deform randomly under the action of external force and increase the toughness.
Description
Technical Field
The invention relates to the technical field of liposome, in particular to a preparation formula and a preparation process of liposome freeze-dried powder.
Background
Since the rapid development of natural science in the 21 st century, people find that more and more medicaments have great help to human health, and the pharmaceutical field also meets the problems, wherein the solubility, stability and transdermal property of the medicaments are one of the key points and difficulties of the current preparation technology. Thus, the carrier technology has become an important issue, wherein the liposome technology is a typical representative of the carrier technology, and the advantages of the liposome technology are as follows: 1. wrapping and bearing the insoluble drug to form a wrapping object which is easily soluble in water; 2. after the unstable components are wrapped, the unstable components are well protected, and the stability of the wrapped objects is improved; 3. the phospholipid bilayer of the liposome has good bionic effect, has good compatibility with the phospholipid bilayer of a cell membrane, such as one track, can greatly improve the transdermal effect, becomes a transdermal carrier, and conveys the medicament to a target position; the disadvantages of liposomes in liquid form are also well defined, they are only relatively stable structures, they are stored for long periods of time and they have a number of unstable factors and phenomena, such as: under the influence of factors such as temperature, temperature difference, illumination, external force and the like, the particle size is enlarged due to mutual fusion, so that the liposome structure is lost, and the encapsulated medicine and the like are released; proliposomes obtained by drying techniques are an important solution to this problem. The significance of liposomes in protecting labile components makes it more desirable to obtain proliposomes by lyophilization techniques.
The freeze-drying technology, which is a deeper preparation drying technology since the 20 th century, has the principle that a medicine water solution containing a certain amount of excipient is frozen into ice through a low-temperature process, so that each water molecule is ensured to exist in an ice state; then, the three-phase line of the medicine is changed in a vacuumizing and heating mode, ice is changed into gaseous water molecules in a sublimation mode all the time and is captured by a cold well, and therefore the purpose of drying is achieved; the whole drying process is carried out in a low-temperature environment, so that the heat-sensitive medicine and the phospholipid bilayer are effectively protected. However, the freeze-dried liposome preparation has many difficulties, such as that due to the change of ice water volume, phospholipid bilayer is destroyed, so that proliposome cannot be formed after freeze-drying, liposome cannot be formed after rehydration, or liposome with larger particle size is formed, and the coating rate of the encapsulated substance is greatly reduced, and the transdermal effect is seriously damaged.
The invention solves the problems in liposome freeze-drying from the innovation of prescription, process and other aspects, and prepares the liposome freeze-dried preparation which has the advantages that the particle size of the liposome after rehydration is close to the particle size before freeze-drying, the coating rate is basically kept unchanged, and the transdermal effect is obvious.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a preparation formula and a preparation process of liposome freeze-dried powder, so as to solve the problems in the background technology.
The technical problem solved by the invention is realized by adopting the following technical scheme: a preparation formula of liposome freeze-dried powder comprises: liposomes, excipients, water; the specific formula is prepared from the following raw materials in percentage by weight: 50% of liposome, 5-15% of excipient and the balance of water; the composition of the liposome is as follows: phospholipid 1-20%, cholesterol 0.1-10%, Tween 0.2-2%, trehalose 5-10%, active substance 0.2-20%, anhydrous ethanol 2-30% and water in balance.
Preferably, the excipient comprises trehalose and one or more of mannitol, sucrose, rhamnose and glucosamine.
Preferably, the tween is one or more of tween 20, tween 60 and tween 80.
Preferably, the active substance is one or more of oligopeptide, resveratrol, sodium ascorbate, tranexamic acid, glabridin, asiaticoside, glycyrrhetinic acid, arbutin, etc.
A preparation process of liposome freeze-dried powder specifically comprises the following steps:
step 1: dissolving active substances, phospholipid and cholesterol in absolute ethyl alcohol according to the raw material percentage of a liposome formula in the liposome freeze-dried preparation to form an ethanol phase; dissolving trehalose, active substances and tween in water to form a water phase;
step 2: injecting the ethanol phase into the water phase at the temperature of 80-95 ℃ at the speed of 0.5-5L/min while stirring, wherein the stirring speed is 30-90 r/min;
and step 3: distilling the uniformly mixed solution obtained in the step 2 under the conditions of 10000Pa of 1000-;
and 4, step 4: according to the raw material percentage of the liposome freeze-dried preparation, uniformly mixing the liposome prepared in the step 3, an excipient and water to obtain freeze-dried mother liquor;
and 5: the freeze-drying mother liquor is respectively filled in a freeze-drying packing material, the temperature is quickly reduced to minus 30 ℃ to minus 40 ℃ at the speed of 1-5 ℃/minute, the temperature is maintained for 30 minutes, the temperature is increased to minus 15 ℃ to minus 20 ℃ at the speed of 1-3 ℃/minute, the temperature is maintained for 5-10 minutes, the temperature is reduced to minus 40 ℃ to minus 50 ℃ at the speed of 0.5-2 ℃/minute, the temperature is maintained for 30 minutes, the pressure in a box is controlled under the vacuum condition of 5-250Pa, the temperature is increased and the mother liquor is sublimated until being dried.
Preferably, the packing material in step 4 is a penicillin bottle, a lyophilized ampoule, a lyophilized blister, or the like.
The application method of the liposome freeze-dried powder is as follows:
the liposome freeze-dried powder can be added into various skin-care external preparations, and the two components are added by 0.1-10 percent to play corresponding active effects, or directly dissolved by a solvent and then smeared on the skin for use.
Compared with the prior art, the invention has the following advantages: the liposome of the invention greatly improves the flexibility of the liposome due to the existence of the Tween, and can lead the liposome to deform randomly under the action of external force, thereby increasing the toughness; the liposome is formed by the participation of trehalose, has the effect of supplementing the position of water molecules in the freeze-drying process, and can be maximally protected in the drying process; the particle size change of the freeze-dried proliposome is within a controllable range after rehydration; the produced liposome freeze-dried powder can be directly added with a solvent for use.
Drawings
FIG. 1 is a graph comparing particle size data of liposomes obtained in accordance with the present invention before and after lyophilization;
FIG. 2 is a graph showing the increase in particle size of a lyophilized powder obtained in an example of the present invention after reconstitution;
fig. 3 is a comparison graph of data from a freeze-dried reconstituted transdermal test obtained in an example of the present invention.
Detailed Description
In order to make the technical means, the creation features, the work flow and the using method of the present invention easily understand and understand the purpose and the efficacy, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, but not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
The formulation of a liposome freeze-dried preparation comprises: liposome, excipient and water, wherein the excipient is mannitol; the specific formula is prepared from the following raw materials in percentage by weight: 5% of mannitol, 50% of liposome and the balance of water; wherein the liposome comprises the following components: 20% of phospholipid, 10% of cholesterol, 2% of tween 20, 5% of trehalose, 7.8% of retinol, 0.2% of glabridin, 10% of ascorbic acid, 2% of tranexamic acid, 30% of absolute ethyl alcohol and water until the water content reaches 100%.
A preparation process of liposome freeze-dried powder specifically comprises the following steps:
step 1: dissolving retinol, glabridin, phospholipid and cholesterol in absolute ethyl alcohol according to the raw material percentage of the liposome formula in the liposome freeze-dried preparation to form an ethanol phase; dissolving trehalose, tranexamic acid, ascorbic acid and tween 20 in water to form a water phase;
step 2: injecting the ethanol phase into the water phase at the temperature of 80 ℃ at the speed of 0.5 liter/min, stirring while injecting, and stirring at the speed of 30 revolutions/min;
and step 3: distilling the uniformly mixed solution obtained in the step 2 under the conditions of 1000Pa and 80 ℃ to remove ethanol; then homogenizing under high pressure, homogenizing under 1200MPa, and controlling particle diameter to 60nm to obtain liposome;
step 4, according to the raw material percentage of the liposome freeze-dried preparation, uniformly mixing the liposome prepared in the step 3, mannitol and water to obtain freeze-dried mother liquor;
and 5: and (3) subpackaging the freeze-dried mother solution in a freeze-drying packaging material, wherein the packaging material is a penicillin bottle, a freeze-dried ampoule bottle or a freeze-dried blister, rapidly reducing the temperature to-30 ℃ at the speed of 1 ℃/minute, preserving the temperature for 30 minutes, raising the temperature to-15 ℃ at the speed of 1 ℃/minute, preserving the temperature for 10 minutes, reducing the temperature to-40 ℃ at the speed of 0.5 ℃/minute, preserving the temperature for 30 minutes, controlling the pressure in the box under the vacuum condition of 5Pa, and raising the temperature for sublimation until the freeze-dried mother solution is dried.
Example 2
The formulation of a liposome freeze-dried preparation comprises: liposome, excipient and water, wherein the excipient is trehalose; the specific formula is prepared from the following raw materials in percentage by weight: 10% of trehalose, 50% of liposome and the balance of water; wherein the liposome comprises the following components: phospholipid 1%, cholesterol 0.5%, Tween-60 0.2%, trehalose 10%, glabridin 0.2%, anhydrous ethanol 2% and water to 100%.
A process for preparing a liposome freeze-dried preparation specifically comprises the following steps:
step 1: dissolving glabridin, phospholipid and cholesterol in absolute ethyl alcohol according to the raw material percentage of a liposome formula in a liposome freeze-dried preparation to form an ethanol phase; dissolving trehalose and tween 60 in water to form a water phase;
step 2: injecting the ethanol phase into the water phase at the temperature of 80 ℃ at the speed of 0.5 liter/min, stirring while injecting, and stirring at the speed of 30 revolutions/min;
and step 3: distilling the uniformly mixed solution obtained in the step 2 under the conditions of 10000Pa and 80 ℃ to remove ethanol; then homogenizing under high pressure, homogenizing under 800MPa, and controlling particle diameter to 150nm to obtain liposome;
step 4, according to the raw material percentage of the liposome freeze-drying preparation, uniformly mixing the liposome, trehalose and water obtained in the step 3 to obtain freeze-drying mother liquor;
and 5: and (3) subpackaging the freeze-dried mother solution in a freeze-drying packaging material, wherein the packaging material is a penicillin bottle, a freeze-dried ampoule bottle or a freeze-dried blister, rapidly reducing the temperature to-40 ℃ at the speed of 1 ℃/minute, preserving the temperature for 30 minutes, raising the temperature to-20 ℃ at the speed of 3 ℃/minute, preserving the temperature for 5 minutes, reducing the temperature to-50 ℃ at the speed of 1 ℃/minute, preserving the temperature for 30 minutes, controlling the pressure in the box under the vacuum condition of 250Pa, and raising the temperature for sublimation until the freeze-dried mother solution is dried.
Example 3
The formulation of a liposome freeze-dried preparation comprises: the liposome, an excipient and water, wherein the excipient is erythritol; the specific formula is prepared from the following raw materials in percentage by weight: 15% of erythritol, 50% of liposome and the balance of water; wherein the liposome comprises the following components: 10% of phospholipid, 5% of cholesterol, 1% of tween 80, 7.5% of trehalose, 0.2% of oligopeptide, 1% of resveratrol, 0.2% of glabridin, 0.5% of vitamin E, 20% of absolute ethyl alcohol and 100% of water.
A process for preparing a liposome freeze-dried preparation specifically comprises the following steps:
step 1: dissolving resveratrol, ceramide, glabridin, phospholipid and cholesterol in absolute ethyl alcohol according to the raw material percentage of a liposome formula in a liposome freeze-dried preparation to form an ethanol phase; dissolving oligopeptide, trehalose and tween 80 in water to form a water phase;
step 2: injecting the ethanol phase into the water phase at the temperature of 80 ℃ at the speed of 0.5 liter/min, stirring while injecting, and stirring at the speed of 30 revolutions/min;
and step 3: distilling the uniformly mixed solution obtained in the step 2 under the conditions of 10000Pa and 80 ℃ to remove ethanol; then homogenizing under high pressure, homogenizing under 800MPa, controlling particle diameter to 120nm, and adding mannitol to obtain liposome;
step 4, according to the raw material percentage of the liposome freeze-dried preparation, uniformly mixing the liposome, erythritol, rhamnose and water obtained in the step 3 to obtain freeze-dried mother liquor;
and 5: and (3) subpackaging the freeze-dried mother solution in a freeze-drying packaging material, wherein the packaging material is a penicillin bottle, a freeze-dried ampoule bottle or a freeze-dried blister, rapidly reducing the temperature to-35 ℃ at the speed of 3 ℃/min, preserving the temperature for 30min, raising the temperature to-18 ℃ at the speed of 2 ℃/min, preserving the temperature for 8 min, reducing the temperature to-45 ℃ at the speed of 0.75 ℃/min, preserving the temperature for 30min, controlling the pressure in the box under the vacuum condition of 150Pa, and heating and sublimating until the box is dried.
Example 4
The formulation of a liposome freeze-dried preparation comprises: the liposome, an excipient and water, wherein the excipient is glucosamine and rhamnose in a mass ratio of 1: 1; the specific formula is prepared from the following raw materials in percentage by weight: 10% of excipient, 50% of liposome and the balance of water; wherein the liposome comprises the following components: 10% of phospholipid, 5% of cholesterol, 1% of tween 80, 7.5% of trehalose, 0.2% of ceramide, 0.2% of glabridin, 0.5% of vitamin E, 20% of absolute ethyl alcohol and water until the water content reaches 100%.
A process for preparing a liposome freeze-dried preparation specifically comprises the following steps:
step 1: dissolving glabridin, ceramide, vitamin E, phospholipid and cholesterol in absolute ethyl alcohol according to the raw material percentage of a liposome formula in a liposome freeze-dried preparation to form an ethanol phase; dissolving trehalose and tween 80 in water to form a water phase;
step 2: injecting the ethanol phase into the water phase at the temperature of 80 ℃ at the speed of 0.5 liter/min, stirring while injecting, and stirring at the speed of 30 revolutions/min;
and step 3: distilling the uniformly mixed solution obtained in the step 2 under the conditions of 10000Pa and 80 ℃ to remove ethanol; then homogenizing under high pressure, homogenizing under 800MPa, and controlling particle diameter to 150nm to obtain liposome;
step 4, according to the raw material percentage of the liposome freeze-drying preparation, uniformly mixing the liposome, the glucosamine, the rhamnose and the water obtained in the step 3 to obtain freeze-drying mother liquor;
and 5: and (3) subpackaging the freeze-dried mother solution in a freeze-drying packaging material, wherein the packaging material is a penicillin bottle, a freeze-dried ampoule bottle or a freeze-dried blister, rapidly reducing the temperature to-35 ℃ at the speed of 3 ℃/min, preserving the temperature for 30min, raising the temperature to-18 ℃ at the speed of 2 ℃/min, preserving the temperature for 8 min, reducing the temperature to-45 ℃ at the speed of 0.75 ℃/min, preserving the temperature for 30min, controlling the pressure in the box under the vacuum condition of 150Pa, and heating and sublimating until the box is dried.
Comparative example 1
The formulation of a liposome freeze-dried preparation comprises: liposome, excipient and water, wherein the excipient is mannitol; the specific formula is prepared from the following raw materials in percentage by weight: 5% of excipient, 50% of liposome and the balance of water; wherein the liposome comprises the following components: 20% of phospholipid, 10% of cholesterol, 5% of trehalose, 7.8% of retinol, 0.2% of glabridin, 10% of ascorbic acid, 2% of tranexamic acid, 30% of absolute ethyl alcohol and water until the water content reaches 100%.
A process for preparing a liposome freeze-dried preparation specifically comprises the following steps:
step 1: dissolving retinol, glabridin, phospholipid and cholesterol in absolute ethyl alcohol according to the raw material percentage of the liposome formula in the liposome freeze-dried preparation to form an ethanol phase; dissolving trehalose, tranexamic acid and ascorbic acid in water to form a water phase;
step 2: injecting the ethanol phase into the water phase at the temperature of 80 ℃ at the speed of 0.5 liter/min, stirring while injecting, and stirring at the speed of 30 revolutions/min;
and step 3: distilling the uniformly mixed solution obtained in the step 2 under the conditions of 1000Pa and 80 ℃ to remove ethanol; then homogenizing under high pressure, homogenizing under 1200MPa, and controlling particle diameter to 60nm to obtain liposome;
step 4, according to the raw material percentage of the liposome freeze-drying preparation, uniformly mixing the liposome, mannitol and water obtained in the step 3 to obtain freeze-drying mother liquor;
and 5: and (3) subpackaging the freeze-dried mother solution in a freeze-drying packaging material, wherein the packaging material is a penicillin bottle, a freeze-dried ampoule bottle or a freeze-dried blister, rapidly reducing the temperature to-30 ℃ at the speed of 1 ℃/minute, preserving the temperature for 30 minutes, raising the temperature to-15 ℃ at the speed of 1 ℃/minute, preserving the temperature for 10 minutes, reducing the temperature to-40 ℃ at the speed of 0.5 ℃/minute, preserving the temperature for 30 minutes, controlling the pressure in the box under the vacuum condition of 5Pa, and raising the temperature for sublimation until the freeze-dried mother solution is dried.
Comparative example 2
The formulation of a liposome freeze-dried preparation comprises: liposome, excipient and water, wherein the excipient is trehalose; the specific formula is prepared from the following raw materials in percentage by weight: 15% of excipient, 50% of liposome and the balance of water; wherein the liposome comprises the following components: phospholipid 1%, cholesterol 0.5%, Tween-60 0.2%, glabridin 0.2%, anhydrous alcohol 2% and water to 100%.
A process for preparing a liposome freeze-dried preparation specifically comprises the following steps:
step 1: dissolving glabridin, phospholipid and cholesterol in absolute ethyl alcohol according to the raw material percentage of a liposome formula in a liposome freeze-dried preparation to form an ethanol phase; dissolving Tween 60 in water to form a water phase;
step 2: injecting the ethanol phase into the water phase at the temperature of 80 ℃ at the speed of 0.5 liter/min, stirring while injecting, and stirring at the speed of 30 revolutions/min;
and step 3: distilling the uniformly mixed solution obtained in the step 2 under the conditions of 10000Pa and 80 ℃ to remove ethanol; then homogenizing under high pressure, homogenizing under 800MPa, and controlling particle diameter to 150nm to obtain liposome;
step 4, according to the raw material percentage of the liposome freeze-drying preparation, uniformly mixing the liposome, trehalose and water obtained in the step 3 to obtain freeze-drying mother liquor;
and 5: and (3) subpackaging the freeze-dried mother solution in a freeze-drying packaging material, wherein the packaging material is a penicillin bottle, a freeze-dried ampoule bottle or a freeze-dried blister, rapidly reducing the temperature to-40 ℃ at the speed of 1 ℃/minute, preserving the temperature for 30 minutes, raising the temperature to-20 ℃ at the speed of 3 ℃/minute, preserving the temperature for 5 minutes, reducing the temperature to-50 ℃ at the speed of 1 ℃/minute, preserving the temperature for 30 minutes, controlling the pressure in the box under the vacuum condition of 250Pa, and raising the temperature for sublimation until the freeze-dried mother solution is dried.
Comparative example 3
The formulation of a liposome freeze-dried preparation comprises: the liposome, an excipient and water, wherein the excipient is erythritol; the specific formula is prepared from the following raw materials in percentage by weight: 15% of excipient, 50% of liposome and the balance of water; wherein the liposome comprises the following components: 10% of phospholipid, 5% of cholesterol, 1% of tween 80, 7.5% of trehalose, 0.2% of oligopeptide, 1% of resveratrol, 0.2% of glabridin, 0.5% of vitamin E, 20% of absolute ethyl alcohol and 100% of water.
A process for preparing a liposome freeze-dried preparation specifically comprises the following steps:
step 1: dissolving resveratrol, ceramide, glabridin, phospholipid and cholesterol in absolute ethyl alcohol according to the raw material percentage of a liposome formula in a liposome freeze-dried preparation to form an ethanol phase; dissolving oligopeptide, trehalose and tween 80 in water to form a water phase;
step 2: injecting the ethanol phase into the water phase at the temperature of 80 ℃ at the speed of 0.5 liter/min, stirring while injecting, and stirring at the speed of 30 revolutions/min;
and step 3: distilling the uniformly mixed solution obtained in the step 2 under the conditions of 10000Pa and 80 ℃ to remove ethanol; then homogenizing under high pressure, homogenizing under 800MPa, controlling particle diameter to 120nm, and adding mannitol to obtain liposome;
step 4, according to the raw material percentage of the liposome freeze-dried preparation, uniformly mixing the liposome, erythritol, rhamnose and water obtained in the step 3 to obtain freeze-dried mother liquor;
and 5: and (3) subpackaging the freeze-dried mother solution in a freeze-drying packaging material, wherein the packaging material is a penicillin bottle, a freeze-dried ampoule bottle or a freeze-dried blister, rapidly reducing the temperature to-35 ℃ at the speed of 3 ℃/min, preserving the heat for 30min, reducing the temperature to-45 ℃, preserving the heat for 30min, controlling the pressure in the box under the vacuum condition of 150Pa, heating to sublimate, and drying.
Comparative example 4
The formulation of a liposome freeze-dried preparation comprises: the liposome, an excipient and water, wherein the excipient is rhamnose; the specific formula is prepared from the following raw materials in percentage by weight: excipient 12%, liposome 50% and water in balance; wherein the liposome comprises the following components: 4% of phospholipid, 2% of cholesterol, 0.2% of tween 80, 6% of trehalose, 0.5% of asiaticoside, 0.2% of glycyrrhetinic acid, 0.2% of glabridin, 2% of salicylic acid, 25% of absolute ethanol and water to 100%.
A process for preparing a liposome freeze-dried preparation specifically comprises the following steps:
step 1: dissolving asiaticoside, glycyrrhetinic acid, salicylic acid, glabridin, phospholipid and cholesterol in absolute ethyl alcohol to form an ethanol phase; dissolving trehalose and tween 80 in water to form a water phase;
step 2: injecting the ethanol phase into the water phase at the temperature of 95 ℃ at the speed of 5 liters/minute, stirring while injecting, and stirring at the speed of 90 revolutions/minute;
and step 3: distilling the uniformly mixed solution obtained in the step 2 at 8000Pa and 85 ℃ to remove ethanol; then homogenizing under high pressure, homogenizing under 1000MPa, and controlling particle diameter to 60nm to obtain liposome;
step 4, according to the raw material percentage of the liposome freeze-dried preparation, uniformly mixing the liposome, the excipient and the water obtained in the step 3 to obtain freeze-dried mother liquor;
and 5: and (3) subpackaging the freeze-dried mother solution in a freeze-drying packaging material, wherein the packaging material is a penicillin bottle, a freeze-dried ampoule bottle or a freeze-dried blister, reducing the temperature to-35 ℃ at the speed of 0.5 ℃/minute, preserving the heat for 30 minutes, raising the temperature to-18 ℃ at the speed of 0.5 ℃/minute, preserving the heat for 8 minutes, reducing the temperature to-45 ℃ at the speed of 2 ℃/minute, preserving the heat for 30 minutes, controlling the pressure in the box under the vacuum condition of 150Pa, raising the temperature and sublimating until the temperature is dried.
Effect example 1: particle size change of liposome lyophilized preparation after rehydration
First, experiment method
The samples of each example and comparative example were subjected to particle size testing using a MS2000 Malvern particle sizer, before and after lyophilization.
Procedure for the preparation of the
1. Instrument start-up
(1) Firstly, opening a computer connected with the MS2000 Marvin particle analyzer, and entering a Windows operating environment.
(2) And opening a host switch of the particle analyzer and a power switch of a disperser of the particle analyzer.
(3) And (4) opening Mastersizer 2000 operating software, entering a main working program by the instrument, and starting up and preheating for 30 min.
2. Parameter setting
(1) The rotation speed of the pump is set, and if necessary, the intensity and time of the ultrasound are set, and about 600ml of a dispersion medium (usually distilled water) is added to a beaker, and then a circulation pump is turned on to circulate the dispersion medium.
(2) And a file is newly created, a manual button in a measurement menu is clicked, and a measurement window is accessed. And then enters the "options" menu.
(3) Clicking a 'material' button, selecting 'sample material' and 'dispersing agent' according to the refractive index, clicking a 'model' to select 'universal mode' to measure an unknown sample; clicking "high", the selection result is converted to "number".
(4) Click the "measure" button and enter "sample measurement time" and "background measurement time". Click "measure" in the high level options, enter the opacity bound "lower bound" and "upper bound".
(5) Click on the "measurement cycle", select the number of measurement cycles and the delay time between measurements, and create an average.
(6) Click "document" to enter sample name "ok".
3. Sample measurement
(1) Clicking the "start" button, the system automatically lights up, measuring the background.
(2) After the background measurement is completed and the 'adding sample' is prompted, the sample is added to the set light shading limit, then the 'start' is clicked, and the instrument measures the sample.
(3) Analysis of results
The sample name was checked on a single click into the results analysis interface and the red d (0.5) is the sample size.
II, experimental results:
the results are shown in FIG. 1 and FIG. 2;
thirdly, analyzing results:
as can be seen from FIGS. 1 and 2, the particle size of the liposome is increased after the rehydration after the lyophilization, wherein the increase of the particle size of the examples is far less than that of the comparative examples, which shows that the prescription and the process of the examples have good stabilizing effect on the liposome lyophilized preparation.
Effect example 2: comparison of transdermal absorption of liposome lyophilized preparation after reconstitution
The first experiment method comprises the following steps:
1. experimental grouping and processing
The liposome freeze-dried preparations of example 1, example 2, example 3, example 4, comparative example 1, comparative example 2, comparative example 3 and comparative example 4 were re-dissolved with water to prepare liposomes containing 500ppm of glabridin for transdermal test, and 500ppm glabridin suspension was used as a control.
2. Preparation of isolated mouse skin
Removing back fur of Kunming qualified mouse with 10% Na2S, removing cervical vertebra, immediately peeling back skin, removing subcutaneous fat layer, cleaning with distilled water, and soaking in distilled water for use.
3. Transdermal absorption test
The skin was fixed between the dosing chamber and the receiving chamber of a diffusion cell using a Franz laboratory apparatus, the effective contact area of the two cells was 2.06cm2, the dermal side was in contact with the receiving fluid, the combination was noted to be tight, no air bubbles were present, and the receiving fluid was 5ml of saline. 1ml of lipid (glabridin 500ppm) rehydrated in each example was added into the supply tank, the magnetic stirring speed was 100r/min, the water bath temperature was 37. + -. 1 ℃, and after 5 hours, a sample of the receiving tank was taken, and the glabridin content in the receiving chamber was measured after the lipid structure of the sample was destroyed.
II, experimental results:
the results are shown in FIG. 3;
thirdly, analyzing results:
from fig. 3, it can be seen that the liposome lyophilized preparations of the examples and the comparative examples have better transdermal effect than glabridin suspension after rehydration, which indicates that the encapsulation is beneficial to the transdermal absorption of the insoluble substances; compared with the transdermal effect of the freeze-dried preparation of the comparative example after rehydration, the transdermal effect of the freeze-dried preparation of the invention after rehydration is obviously improved.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (6)
1. A preparation formula of liposome freeze-dried powder is characterized in that: the method comprises the following steps: liposomes, excipients, water; the specific formula is prepared from the following raw materials in percentage by weight: 50% of liposome, 5-15% of excipient and the balance of water; the composition of the liposome is as follows: phospholipid 1-20%, cholesterol 0.1-10%, Tween 0.2-2%, trehalose 5-10%, active substance 0.2-20%, anhydrous ethanol 2-30% and water in balance.
2. The preparation formula of the liposome lyophilized powder of claim 1, which is characterized in that: the excipient comprises one or more of trehalose and mannitol, sucrose, rhamnose and glucosamine.
3. The preparation formula of the liposome lyophilized powder of claim 1, which is characterized in that: the tween is one or more of tween 20, tween 60 and tween 80.
4. The preparation formula of the liposome lyophilized powder of claim 1, which is characterized in that: the active substance is one or more of oligopeptide, resveratrol, sodium ascorbate, tranexamic acid, glabridin, asiaticoside, glycyrrizoic acid, arbutin, etc.
5. A preparation process of liposome freeze-dried powder is characterized in that: the method specifically comprises the following steps of,
step 1: dissolving active substances, phospholipid and cholesterol in absolute ethyl alcohol according to the raw material percentage of a liposome formula in the liposome freeze-dried preparation to form an ethanol phase; dissolving trehalose, active substances and tween in water to form a water phase;
step 2: injecting the ethanol phase into the water phase at the temperature of 80-95 ℃ at the speed of 0.5-5L/min while stirring, wherein the stirring speed is 30-90 r/min;
and step 3: distilling the uniformly mixed solution obtained in the step 2 under the conditions of 10000Pa of 1000-;
and 4, step 4: according to the raw material percentage of the liposome freeze-dried preparation, uniformly mixing the liposome prepared in the step 3, an excipient and water to obtain freeze-dried mother liquor;
and 5: the freeze-drying mother liquor is respectively filled in a freeze-drying packing material, the temperature is quickly reduced to minus 30 ℃ to minus 40 ℃ at the speed of 1-5 ℃/minute, the temperature is maintained for 30 minutes, the temperature is increased to minus 15 ℃ to minus 20 ℃ at the speed of 1-3 ℃/minute, the temperature is maintained for 5-10 minutes, the temperature is reduced to minus 40 ℃ to minus 50 ℃ at the speed of 0.5-2 ℃/minute, the temperature is maintained for 30 minutes, the pressure in a box is controlled under the vacuum condition of 5-250Pa, the temperature is increased and the mother liquor is sublimated until being dried.
6. The preparation process of the freeze-dried liposome powder according to claim 5, which is characterized in that: and the packing materials in the step 4 are penicillin bottles, freeze-dried ampoules, freeze-dried bubble caps and the like.
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| CN113713182A (en) * | 2021-09-22 | 2021-11-30 | 苏州大学附属第二医院 | Preparation and application of resveratrol liposome-containing guided bone tissue regeneration membrane |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104721138A (en) * | 2013-12-23 | 2015-06-24 | 天津芸熙生物技术有限公司 | Glabridin freeze-dried liposome and preparation method thereof |
| CN104940040A (en) * | 2015-07-09 | 2015-09-30 | 西安艾尔菲生物科技有限公司 | Liposomes modified with hyaluronic acid and used for encapsulating whitening components, preparation method and application |
| CN104997652A (en) * | 2015-07-09 | 2015-10-28 | 西安艾尔菲生物科技有限公司 | Crease resistance and moisture retention liposome, and preparation method and application thereof |
| US20180338918A1 (en) * | 2017-05-24 | 2018-11-29 | Jiangsu Tasly Diyi Pharmaceutical Co., Ltd. | Temsirolimus liposome and preparation method thereof |
-
2020
- 2020-09-27 CN CN202011030543.XA patent/CN112137968B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104721138A (en) * | 2013-12-23 | 2015-06-24 | 天津芸熙生物技术有限公司 | Glabridin freeze-dried liposome and preparation method thereof |
| CN104940040A (en) * | 2015-07-09 | 2015-09-30 | 西安艾尔菲生物科技有限公司 | Liposomes modified with hyaluronic acid and used for encapsulating whitening components, preparation method and application |
| CN104997652A (en) * | 2015-07-09 | 2015-10-28 | 西安艾尔菲生物科技有限公司 | Crease resistance and moisture retention liposome, and preparation method and application thereof |
| US20180338918A1 (en) * | 2017-05-24 | 2018-11-29 | Jiangsu Tasly Diyi Pharmaceutical Co., Ltd. | Temsirolimus liposome and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| 唐星 等: "《药剂学》", 31 December 2019, 中国医药科技出版社 * |
| 王恒通 等: "《药厂GMP应知应会》", 31 July 2019, 中国医药科技出版社 * |
| 金谷 等: "《表面活性剂化学》", 31 August 2013, 中国科学技术大学出版社 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113713182A (en) * | 2021-09-22 | 2021-11-30 | 苏州大学附属第二医院 | Preparation and application of resveratrol liposome-containing guided bone tissue regeneration membrane |
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