CN102210870A - Liposome composite phospholipid capable of adjusting phase-transition temperature and application thereof - Google Patents
Liposome composite phospholipid capable of adjusting phase-transition temperature and application thereof Download PDFInfo
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- liposome
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- 238000000034 method Methods 0.000 claims description 22
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- QMGVPVSNSZLJIA-UHFFFAOYSA-N Nux Vomica Natural products C1C2C3C4N(C=5C6=CC=CC=5)C(=O)CC3OCC=C2CN2C1C46CC2 QMGVPVSNSZLJIA-UHFFFAOYSA-N 0.000 claims description 12
- 241001279009 Strychnos toxifera Species 0.000 claims description 12
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- 239000006185 dispersion Substances 0.000 claims description 4
- JGSARLDLIJGVTE-UHFFFAOYSA-N 3,3-dimethyl-7-oxo-6-[(2-phenylacetyl)amino]-4-thia-1-azabicyclo[3.2.0]heptane-2-carboxylic acid Chemical compound O=C1N2C(C(O)=O)C(C)(C)SC2C1NC(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-UHFFFAOYSA-N 0.000 claims description 3
- 239000000499 gel Substances 0.000 claims description 3
- 239000000725 suspension Substances 0.000 claims description 3
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 2
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 2
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- 229950004354 phosphorylcholine Drugs 0.000 abstract 1
- DEGAKNSWVGKMLS-UHFFFAOYSA-N calcein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(CN(CC(O)=O)CC(O)=O)=C(O)C=C1OC1=C2C=C(CN(CC(O)=O)CC(=O)O)C(O)=C1 DEGAKNSWVGKMLS-UHFFFAOYSA-N 0.000 description 17
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- LYPFDBRUNKHDGX-SOGSVHMOSA-N N1C2=CC=C1\C(=C1\C=CC(=N1)\C(=C1\C=C/C(/N1)=C(/C1=N/C(/CC1)=C2/C1=CC(O)=CC=C1)C1=CC(O)=CC=C1)\C1=CC(O)=CC=C1)C1=CC(O)=CC=C1 Chemical compound N1C2=CC=C1\C(=C1\C=CC(=N1)\C(=C1\C=C/C(/N1)=C(/C1=N/C(/CC1)=C2/C1=CC(O)=CC=C1)C1=CC(O)=CC=C1)\C1=CC(O)=CC=C1)C1=CC(O)=CC=C1 LYPFDBRUNKHDGX-SOGSVHMOSA-N 0.000 description 7
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- 229910001868 water Inorganic materials 0.000 description 3
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
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- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
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- WPEVAYGXMBMZTM-UHFFFAOYSA-N CCCCCCCCCCCCCCCCCCC(CCCCCCCCCCCCCCCCCC)(C[N+](C)(C)C)OP([O-])(O)=O Chemical compound CCCCCCCCCCCCCCCCCCC(CCCCCCCCCCCCCCCCCC)(C[N+](C)(C)C)OP([O-])(O)=O WPEVAYGXMBMZTM-UHFFFAOYSA-N 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
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- 229930003427 Vitamin E Natural products 0.000 description 1
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- 244000309466 calf Species 0.000 description 1
- WORJEOGGNQDSOE-UHFFFAOYSA-N chloroform;methanol Chemical class OC.ClC(Cl)Cl WORJEOGGNQDSOE-UHFFFAOYSA-N 0.000 description 1
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- BNIILDVGGAEEIG-UHFFFAOYSA-L disodium hydrogen phosphate Chemical compound [Na+].[Na+].OP([O-])([O-])=O BNIILDVGGAEEIG-UHFFFAOYSA-L 0.000 description 1
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- 238000004090 dissolution Methods 0.000 description 1
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- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
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Abstract
The invention discloses liposome composite phospholipid capable of adjusting phase-transition temperature and application thereof. The liposome composite phospholipid is prepared from 1-10 molar parts of dipalmitoyl phosphorylcholine (DPPC) and 1-10 molar parts of hydrogenated soya phosphatidylcholine (HSPC), and can be used for preparing thermosensitive target liposome preparation. According to the invention, different phospholipid materials are screened through a large number of experiments, and the experimental result indicates that the liposome composite phospholipid capable of adjusting phase-transition temperature can be obtained by scientifically combining DPPC and HSPC; the composite phospholipid can form a new phase when being prepared into a liposome membrane, avoid phase separation, has a single phase-transition temperature and can overcome the shortcoming of the prior art that two different phospholipid materials have two phase-transition temperatures; and moreover, the composite phospholipid has better stability and target release effect compared with the single-DPPC liposome, has phase-transition temperature with better tolerance to human body compared with the single-HSPC liposome, and has wide application range.
Description
Technical field
The present invention relates to technical field of medicine, be specifically related to a kind ofly can regulate the liposome composite phospholipid of phase transition temperature and the application in pharmaceutical preparation thereof.
Background technology
Phase transition temperature (phase transition temperature, T
m) be one of most important physicochemical characteristics parameter of liposome membrane, when temperature reaches phase transition temperature, liposome membrane is that phospholipid bilayer has become unordered liquid crystalline phase (liquid disordered phase) from solid crystalline phase (solid gel phase), C-C singly-bound in the phospholipid alkyl chain is the skew form conformation by the alltrans conformation transition, cause the shared cumulative volume of hydrocarbon chain to increase considerably, thereby cause the liposome membrane permeability sharply to increase thereupon, medicine from liposome fast, fully discharge.The height of phase change temperature of liposome depends on the phospholipid kind that is used to prepare liposome.
Use phase transition temperature principle design thermal sensitive liposome, be called the responsive to temperature liposome again, its used phospholipid material is often selected DPPA choline (DPPC) for use, phase transition temperature is 41 ~ 42 ℃.Behind the intravenous injection medicine carrying thermal sensitive liposome, target site is heated to can realizes more than the liposome membrane phase transition temperature that targeting discharges medicine: the medicine carrying thermal sensitive liposome arrives before the target site under 37 ℃ of body temperature (being lower than phase transition temperature), liposome membrane is in solid crystalline phase, permeability is low, and medicine does not leak substantially; And the thermal sensitive liposome of arrival target site, the liposome membrane temperature is heated to more than the phase transition temperature, thereby rapidly, fully discharges medicine at target site selectively.
At present because the phospholipid of phase transition temperature between 40 ~ 50 ℃ of scopes in has only DPPA choline (DPPC), so it is the phospholipid material of present thermal sensitive liposome preparation indispensability, but the price comparison costliness.Temperature is higher than 50 ℃ distearyl phosphocholine (DSPC), hydrogenated soya phosphatide phospholipid materials such as (HSPC) can't be as the main phospholipid material of thermal sensitive liposome, because if can aggravate heating-up temperature damage to most of normal structures and physiological function above 50 ℃ of target site.
The matrix material of thermal sensitive liposome is formed except that cholesterol, targeting modification material (as stealthy decorative material DSPE-Macrogol 2000), generally only be made up of a kind of phospholipid, phase transition temperature depends primarily on the phase transition temperature that this phospholipid material possesses.But the adding of medicine has often changed the phase transition temperature of drug-loaded liposome again, for example add for behind the Mo Luofen (Temoporfin), the phase transition temperature of DPPA choline (DPPC) liposome significantly descends, even be lower than 37 ℃ of body temperature (Kuntsche J, Freisleben I, Steiniger F, et al. Temoporfin-loaded liposomes:physicochemical characterization [J]. European Journal of Pharmaceutical Sciences, 2010,40:305-315.).Therefore be badly in need of a kind of phospholipid material that can the flexible phase change temperature of liposome, medicine behind the medicine carrying can be eliminated the influence of phase change temperature of liposome, thereby guarantee the targeting drug release effect of thermal sensitive liposome.
In addition, DPPC thermal sensitive liposome stable limited under 37 ℃ of body temperature, the part medicine just leaks under 37 ℃ of body temperature, thereby reduction curative effect of medication, therefore the necessary screening of research on the basis of existing technology is a kind of can improve thermal sensitive liposome under body temperature stability and don't influence its release under phase transition temperature, can significantly reduce the liposome phospholipid of the cost of thermal sensitive liposome simultaneously again.
But, form the preparation liposome if use two or more phospholipid materials simultaneously, in most cases, different phospholipid still keep specific separately phase transition temperature in liposome membrane, when temperature is between the phase transition temperature of two kinds of phospholipid, have different phases in the liposome membrane simultaneously, be called be separated (phase separations), cause the film surface to form block structure (domain structure), the permeability of lipid film is increased, cause drug leakage.(the Lu Bin chief editor, novel pharmaceutical formulation and new technique (the 2nd edition), the People's Health Publisher, 2005:127-128).
Summary of the invention
Goal of the invention: the objective of the invention is in order to solve the deficiencies in the prior art, a kind of liposome composite phospholipid that can regulate phase transition temperature is provided, this liposome composite phospholipid is compared with existing DPPC thermal sensitive liposome, can the flexible phase transition temperature, improve the stability of liposome under body temperature, reduce cost simultaneously; Another object of the present invention provides the application of liposome composite phospholipid in preparation heat sensitive target Liposomal formulation that can regulate phase transition temperature.
Technical scheme: in order to realize above purpose, the technical scheme that the present invention takes is:
A kind of liposome composite phospholipid that can regulate phase transition temperature, it is to be made by the raw material of following molfraction:
1 ~ 10 part in DPPA choline, 1 ~ 10 part of hydrogenated soya phosphatide.
As preferred version, the above-described liposome composite phospholipid that can regulate phase transition temperature, it is to be made by the raw material of following mole ratio:
The DPPA choline: hydrogenated soya phosphatide equals 4:1 to 1:4.
As more preferably scheme, the above-described liposome composite phospholipid that can regulate phase transition temperature, it is to be made by the raw material of following mole ratio:
The DPPA choline: hydrogenated soya phosphatide equals 2:8,4:6,5:5,6:4 or 8:2.
A kind of mathematical function that is used to design the composite phospholipid material liposome of different phase transition temperatures provided by the invention, this mathematical function equation is y * 1000=0.1077x+3.0638, R
2=0.9895, wherein the y axle is the 1/T reciprocal of the absolute temperature of DPPA choline and hydrogenated soya phosphatide liposome composite phospholipid phase transition temperature, and the x axle is DPPA choline shared molar ratio in DPPA choline and hydrogenated soya phosphatide total amount.Utilize this mathematical function relationship can be used to design the liposome of different phase transition temperatures, especially can be used to design the design of medicine the influential liposome of phase change temperature of liposome.
The application of liposome composite phospholipid in preparation heat sensitive target Liposomal formulation that can regulate phase transition temperature provided by the invention.
As preferred version, the above-described application of liposome composite phospholipid in preparation heat sensitive target Liposomal formulation that can regulate phase transition temperature, described heat sensitive target Liposomal formulation is the medicine of injectable powder, injection, oral liquid, exterior-applied gel, ointment, spray or nasal drop dosage form.When preparation said medicine preparation, also can add other adjuvant, as vitamin E, DSPE-Macrogol 2000 (DSPE-PEG2000), sodium chloride, sodium hydrogen phosphate, sodium dihydrogen phosphate, water etc.
Composite phospholipid provided by the invention is formed the heat sensitive target liposome for preparing, and after the depyrogenation, through aseptic filtration, lyophilization obtains injection heat sensitive target liposome powder for injection.
When the heat sensitive target liposome that composite phospholipid composition provided by the invention prepares is made injection, get each raw material adding physiological saline solution and add active carbon then, stir, filter, regulate pH value, be filtered to sintered glass funnel or other filter clear and bright, microporous filter membrane aseptic filtration, injection is made in fill.
When composite phospholipid provided by the invention is formed the heat sensitive target liposome for preparing and made oral liquid,, filter, add syrup, regulate pH value, make oral liquid each raw material adding distil water dissolving.
As preferred version, the above-described preparation method that can regulate the heat sensitive target Liposomal formulation of phase transition temperature is film dispersion method, reverse evaporation, alcohol injection, ether injection method, freeze-drying, ammonium sulphate gradient, pH gradient method or pro-liposome method;
The most preferred scheme, preparation method is a film dispersion method, described film dispersion method may further comprise the steps: (1) takes by weighing 1～10 part of 1～10 part of DPPC, HSPC and fat-soluble medicine, other matrix materials in molar ratio, be dissolved in dehydrated alcohol or other organic solvent, decompression rotary evaporation film forming, vacuum condition is placed down.(2) add aqueous medium (water miscible medicine dissolution is in aqueous medium) rotation hydration, make liposome turbid liquor.(3) supersound process or high pressure homogenize, promptly.
The strychnine composite phospholipid hidden liposome lyophilized injectable powder that can regulate phase transition temperature provided by the invention, it is to prepare by the following method: take by weighing 900mg DPPA choline, the 300mg hydrogenated soya phosphatide, 5mg DSPE-Macrogol 2000, the 100mg strychnine is dissolved in the 50ml dehydrated alcohol altogether, under 60 ℃ bath temperature, decompression rotation film forming, vacuum is placed and is spent the night, the phosphate PBS buffer rotation hydration that adds 50mL pH7.4, aqua liquid extra high pressure homogenize machine homogenize, cross 0.22 μ m microporous filter membrane degerming, be the milky suspension clear and bright to light, be sub-packed in the cillin bottle, elder generation's pre-freeze, put lyophilization in the freezer dryer then, promptly get phase transition temperature and be 44.07 ℃ strychnine composite phospholipid hidden liposome lyophilized injectable powder.
The present invention screens the phospholipid material of two kinds of different phase transition temperatures through lot of experiments: DPPA choline and hydrogenated soya phosphatide, show when forming cenotype when composite phospholipid material preparation that the hydrogenated soya phosphatide of 1 ~ 10 part of molfraction is formed becomes liposome membrane through lot of experiments by the DPPA choline of 1 ~ 10 part of molfraction, do not exist and be separated, especially have single phase transition temperature, overcome the shortcoming of two kinds of phase transition temperatures of two kinds of different phospholipid materials existence of prior art.
Beneficial effect: the liposome composite phospholipid that can regulate phase transition temperature provided by the invention is compared with prior art and is had the following advantages:
1, the present invention is by the different phospholipid material of a large amount of experiment screenings, experimental result shows the liposome composite phospholipid that can obtain regulating phase transition temperature after the hydrogenated soya phosphatide scientific combination of the DPPA choline of 1 ~ 10 part of molfraction and 1 ~ 10 part of molfraction, when being prepared into liposome membrane, this composite phospholipid can form cenotype, do not exist and be separated, have single phase transition temperature, can overcome the shortcoming that there are two kinds of phase transition temperatures in two kinds of different phospholipid materials of prior art;
2, the present invention passes through lot of experiments, determine the phase transition temperature of liposome composite phospholipid and the mathematical function rule between two kinds of phospholipid ratios, can regulate phase transition temperature neatly according to the needs of difference treatment according to this mathematical function, thereby medicine behind the medicine carrying can be eliminated the influence of phase change temperature of liposome, be guaranteed the targeting drug release effect of thermal sensitive liposome;
3, the liposome composite phospholipid that can regulate phase transition temperature provided by the invention, compare with the DPPC liposome that only adopts single phospholipid material, the stability of liposome under 37 ℃ of body temperature significantly strengthens, can significantly reduce the leakage drug-loaded liposome arrives target site under 37 ℃ of body temperature before, thereby strengthen the targeted delivery effect of liposome, and the phase transition temperature of liposome composite phospholipid provided by the invention is lower than the high phase transition temperature of single component HSPC, human body can be well tolerable, and since the not enough DPPC of the price of HSPC 10%, therefore composite phospholipid material provided by the invention can not only significantly improve the stability of liposome membrane, increase liposome targeting drug effect, and can reduce the preparation cost of liposome, have wide range of applications.
4, the strychnine composite phospholipid hidden liposome lyophilized injectable powder that can regulate phase transition temperature provided by the invention, the concrete consumption proportion of composite, strychnine and the stealthy decorative material etc. of a large amount of experiment screening phospholipid of process, can obtain that stability is better, targeting drug release better effects if, Semen Strychni liposome that drug loading is higher, and can make things convenient for drug administration by injection through preparing lyophilized injectable powder.
Description of drawings
Fig. 1 is the releasing curve diagram of calcein liposome under different temperatures that different phospholipid are formed.
Fig. 2 is the release comparative graph of calcein liposome under 37 ℃ that different phospholipid are formed.
The different phospholipid of Fig. 3 are formed the DSC collection of illustrative plates of liposome.
Fig. 4 is the mathematical relationship figure that phase transition temperature and composite phospholipid are formed.
Fig. 5 should be the phase transition temperature measurement result figure of the liposome composite phospholipid of 0.4607:0.5393 for the mol ratio of DPPC and HSPC.
The specific embodiment
Further illustrate the present invention below in conjunction with specific embodiment, should understand these embodiment only is used to the present invention is described and is not used in and limit the scope of the invention, after having read the present invention, those skilled in the art all fall within the application's claims institute restricted portion to the modification of the various equivalent form of values of the present invention.
The preparation of embodiment 1 calcein composite phospholipid liposome and the experiment of release in vitro behavior determination
Press DPPC and HSPC mol ratio 2:8 respectively, 4:6,5:5,6:4 and 8:2 take by weighing each 120 mg of composite phospholipid material and are dissolved in 10 mL chloroforms respectively, place 60 ℃ of decompression rotary evaporations formation thin film in the 50 mL eggplant-shape bottles, take out eggplant-shape bottle, in vacuum drying oven, place 12h for 55 ℃.In eggplant-shape bottle, add pH7.4 phosphate buffer (PBS) 5 mL that contain 90 mM calceins then respectively, 60 ℃ of rotation hydration 40 min, the gained liposome turbid liquor is in 60 ℃ of ultrasonic (400 W of water-bath probe, 50 times), promptly get the calcein composite phospholipid liposome of different mol ratio DPPC and HSPC.Prepare calcein DPPC and the HSPC liposome that only contains a DPPC and a HSPC phospholipid material respectively with method.
Above calcein liposome is removed free drug with the gel filtration chromatography method, with calf serum--pH7.4PBS(1:1, v/v) be release medium, each liposome with release medium dilution back constant temperature magnetic agitation, is measured the release of placing 20min.By the release of different phospholipid composition calcein liposomees under the above method investigation different temperatures, concrete experimental result as shown in Figure 1.
Release experimental result by the calcein liposome of Fig. 1 shows, adopt the temperature of the highest release of calcein composite phospholipid liposome that different mol ratio DPPC and HSPC make to improve regularly along with the raising of HSPC ratio, point out its phase transition temperature to take place correspondingly to improve, the calcein liposome that different mol ratio DPPC and HSPC composite phospholipid material are made all has single phase transition temperature, and phase transition temperature all is higher than the calcein liposome that single DPPC material is made, and equal calcein liposome of making less than single HSPC material, can overcome not only that two kinds of phospholipid keep specific separately phase transition temperature in the prior art in liposome membrane, when temperature is between the phase transition temperature of two kinds of phospholipid, the shortcoming that is separated appears in the liposome membrane, and it is unstable under body temperature to overcome single DPPC material liposome, the slip height causes the shortcoming of targeting drug release weak effect, can overcome the too high shortcoming of single HSPC material phase change temperature of liposome in the prior art again.
The release behavior that 2 37 ℃ of down different phospholipid of embodiment are formed the calcein liposome compares
Getting DPPC and the HSPC mol ratio that embodiment 1 prepares is 2:8,4:6,5:5, the calcein liposome that 6:4 and 8:2 composite phospholipid and single DPPC and HSPC material are made, at the release behavior of 37 ℃ of following above calcein liposomees, concrete experimental result is seen shown in Figure 2 then.After experimental result shows adding certain proportion HSPC, the prepared stability of calcein composite phospholipid liposome under 37 ℃ significantly is better than the calcein liposome of single DPPC phospholipid, and present tangible dependency, the ratio of HSPC is high more, composite phospholipid liposome is stable more, and release is low more.Therefore this experiment shows that further liposome that preferred DPPC of the present invention and HSPC composite phospholipid material are made has all under 37 ℃ of normal body temperatures of human body that to make liposome than single DPPC phospholipid material better stable, can prevent effectively that leaking from appearring in medicine before arriving targeting moiety, thereby have better targeting drug release effect.
Embodiment 3 differential scanning calorimetric analysis methods (DSC) are measured the phase transition temperature of liposome and the correlation research that phospholipid is formed
1, the preparation of liposome: (mol ratio of DPPC and HSPC is respectively 2:8 to take by weighing the composite phospholipid material respectively, 4:6,5:5,6:4,8:2) 120 mg are dissolved in 10 mL chloroforms respectively, place 60 ℃ of decompression rotary evaporations formation thin film in the 50 mL eggplant-shape bottles then, take out eggplant-shape bottle, in vacuum drying oven, place 12h for 55 ℃.In eggplant-shape bottle, add pH7.4 phosphate buffer (PBS) 5 mL, 60 ℃ of rotation hydration 40 min, the gained liposome turbid liquor is in 60 ℃ of ultrasonic (400 W of water-bath probe, 50 times), promptly get different mol ratio example DPPC and HSPC(and be respectively 2:8,4:6,5:5,6:4, the composite phospholipid liposome of 8:2) making.
Prepare DPPC, the HSPC liposome that only contains a DPPC and a HSPC phospholipid material respectively with method.
Get the composite phospholipid liposome of above different proportion and the liposome of single DPPC or HSPC phospholipid composition and carry out differential scanning calorimetric analysis under liquid state, the DSC result of mensuration is (a is that HSPC liposome, b are that HSPC:DPPC=8:2 liposome, c are that HSPC:DPPC=6:4 liposome, d are that HSPC:DPPC=5:5 liposome, e are that HSPC:DPPC=4:6 liposome, f are that HSPC:DPPC=2:8 liposome, g are the DPPC liposome among the figure) as shown in Figure 3.Measurement result by Fig. 3 shows the raising of composite phospholipid liposome along with the HSPC ratio, the phase transition temperature of composite phospholipid liposome presents raising regularly, and two phase transition temperatures do not appear among the DSC mensuration figure, therefore show that the liposome that composite phospholipid material provided by the invention is made has the single temperature of comparing, do not exist and be separated, and show that by measurement result the phase transition temperature of composite phospholipid liposome all is higher than the liposome that single DPPC material is made, and equal liposome of making less than single HSPC material, therefore it is better to have stability, targeting drug release better effects if, phase transition temperature again can too high advantages.
2, the phase transition temperature of the liposome that DSC is recorded (inverse with absolute temperature T is represented) is carried out the mathematics match with DPPC shared molar ratio in (DPPC+HSPC) total amount, and the mathematical function that obtains is: y * 1000=0.1077x+3.0638, R
2=0.9895, the result presents clear regularity between visible 1/T and the DPPC ratio as shown in Figure 4, correlation coefficient reaches 0.9947, have good linear relationship, when utilizing this mathematical function can measure in the range of linearity arbitrary phase transition temperature, the mol ratio of DPPC and HSPC in the composite phospholipid material.
As measuring according to this mathematical function: if obtain phase transition temperature is 48.04 ℃ liposome composite phospholipid, and then the mol ratio of DPPC and HSPC should be 0.4607:0.5393.The liposome that invention should be the 0.4607:0.5393 preparation according to the mol ratio of DPPC and HSPC carries out phase transition temperature and measures, concrete phase transition temperature measurement result as shown in Figure 5, the phase transition temperature of measuring is 47.77 ℃, practical measurement value and theoretical prediction value be deviation-0.56% only, and the reliability and the suitability of mathematical function provided by the invention are described.Utilize this mathematical function relationship can be used to design the liposome of different phase transition temperatures.
Embodiment 4 uses the phase transition temperature that composite phospholipid is regulated drug-loaded liposome
Taking by weighing DPPC 120 mg and temoporfin 10mg is dissolved in 10 mL chloroform-methanols (3:1 v/v), places that 60 ℃ of decompression rotary evaporations form thin film in the 50 mL eggplant-shape bottles, takes out eggplant-shape bottle, places 12h for 55 ℃ in vacuum drying oven.In eggplant-shape bottle, add pH7.4 phosphate buffer (PBS) 5 mL then respectively, 60 ℃ of rotation hydration 40 min, the gained liposome turbid liquor promptly gets temoporfin DPPC liposome in 60 ℃ of water-bath probes ultrasonic (400 W, 50 times).Make the blank DPPC liposome that does not contain for Mo Baifen with method.
Take by weighing DPPC90mg and HSPC30mg and prepare the temoporfin composite phospholipid liposome as stated above as the liposome composite phospholipid.
The phase transition temperature that DSC measures blank DPPC liposome is 41.80 ℃, the phase transition temperature of temoporfin DPPC liposome is 36.22 ℃, and the phase transition temperature of temoporfin composite phospholipid liposome is 41.86 ℃, as seen adopts composite phospholipid to regulate suitable ratio and medicine can be eliminated the influence of phase change temperature of liposome.
The preparation of embodiment 5 strychnine composite phospholipid hidden liposome freeze-dried powders
Take by weighing 900mg DPPC, 300mg HSPC, 5mg DSPE-Macrogol 2000, the 100mg strychnine is dissolved in the 50ml dehydrated alcohol altogether, under 60 ℃ bath temperature, decompression rotation film forming, vacuum is placed and is spent the night, the PBS rotation hydration that adds 50mL pH7.4, aqua liquid extra high pressure homogenize machine homogenize, cross 0.22 μ m microporous filter membrane degerming, be the milky suspension clear and bright to light, be sub-packed in by (2ml/ bottle) in the cillin bottle, elder generation's pre-freeze, put lyophilization in the freezer dryer then, promptly get strychnine composite phospholipid hidden liposome freeze-dried powder, its phase transition temperature of differential scanning calorimetric analysis method (DSC) algoscopy is 44.07 ℃, and this strychnine composite phospholipid hidden liposome has higher phase transition temperature than single phospholipid material such as DPPC liposome, therefore has better stability under the normal human puts question to, and 44 ℃ can tolerate for human body, can overcome the shortcoming of comparing temperature more than 50 ℃ of single HSPC or DSPC material liposome.And the result shows after testing, and the envelop rate of strychnine composite phospholipid hidden liposome provided by the invention reaches 95%, and the liposome that obtains than prior art for preparing has higher drug loading and envelop rate.
The above only is a preferred implementation of the present invention; should be pointed out that for those skilled in the art, under the prerequisite that does not break away from the principle of the invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (8)
1. the liposome composite phospholipid that can regulate phase transition temperature is characterized in that, it is to be made by the raw material of following molfraction:
1 ~ 10 part in DPPA choline, 1 ~ 10 part of hydrogenated soya phosphatide.
2. the liposome composite phospholipid that can regulate phase transition temperature according to claim 1 is characterized in that, it is to be made by the raw material of following mole ratio:
The DPPA choline: hydrogenated soya phosphatide equals 4:1 to 1:4.
3. the liposome composite phospholipid that can regulate phase transition temperature according to claim 1 is characterized in that, it is to be made by the raw material of following mole ratio:
The DPPA choline: hydrogenated soya phosphatide equals 2:8,4:6,5:5,6:4 or 8:2.
4. each the described liposome composite phospholipid application in preparation heat sensitive target Liposomal formulation that can regulate phase transition temperature of claim 1 to 3.
5. the application of liposome composite phospholipid in preparation heat sensitive target Liposomal formulation that can regulate phase transition temperature according to claim 4, it is characterized in that described heat sensitive target Liposomal formulation is the medicine of injectable powder, injection, oral liquid, exterior-applied gel, ointment, spray or nasal drop dosage form.
6. the application of liposome composite phospholipid in preparation heat sensitive target Liposomal formulation that can regulate phase transition temperature according to claim 4, it is characterized in that the preparation method of described heat sensitive target Liposomal formulation is film dispersion method, reverse evaporation, alcohol injection, ether injection method, freeze-drying, ammonium sulphate gradient, pH gradient method or pro-liposome method.
7. a mathematical function that is used to design the composite phospholipid liposome of different phase transition temperatures is characterized in that this mathematical function equation is y * 1000=0.1077x+3.0638, R
2=0.9895, wherein the y axle is the inverse of the absolute temperature of DPPA choline and hydrogenated soya phosphatide liposome composite phospholipid phase transition temperature, and the x axle is DPPA choline shared molar ratio in DPPA choline and hydrogenated soya phosphatide total amount.
8. the strychnine composite phospholipid hidden liposome lyophilized injectable powder that can regulate phase transition temperature, it is characterized in that, it is to prepare by the following method: take by weighing 900mg DPPA choline, the 300mg hydrogenated soya phosphatide, 5mg DSPE-Macrogol 2000, the 100mg strychnine is dissolved in the 50ml dehydrated alcohol altogether, under 60 ℃ bath temperature, decompression rotation film forming, vacuum is placed and is spent the night, the PBS buffer rotation hydration that adds 50mL pH7.4, aqua liquid extra high pressure homogenize machine homogenize, cross 0.22 μ m microporous filter membrane degerming, be the milky suspension clear and bright to light, be sub-packed in the cillin bottle, elder generation's pre-freeze is put lyophilization in the freezer dryer, promptly then.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102688153A (en) * | 2012-06-05 | 2012-09-26 | 东南大学 | Blank liposome prepared by compounding phospholipid and preparation method thereof |
| CN103404844A (en) * | 2013-04-26 | 2013-11-27 | 江南大学 | Composition capable of enhancing stability of lycopene and having function of relieving physical fatigue and preparation method thereof |
| CN104785178A (en) * | 2015-03-26 | 2015-07-22 | 江南大学 | Thermosensitive liposome as well as preparation method and application thereof |
| CN107998070A (en) * | 2017-12-12 | 2018-05-08 | 南京中医药大学 | A kind of composite phospholipid carrier and its application in percutaneous drug administration preparation is prepared |
| CN108030929A (en) * | 2017-12-19 | 2018-05-15 | 浙江农林大学 | A kind of preparation method of Lazer's property liposome comprising berberine |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1883454A (en) * | 2006-06-01 | 2006-12-27 | 复旦大学 | Liposome administration system with tumor necrosis targeted antibody for preposition and application thereof |
| CN101406455A (en) * | 2008-11-28 | 2009-04-15 | 南京中医药大学 | Strychnine composite phospholipid liposome, and preparation method thereof and use in pharmacy |
| CN101744767A (en) * | 2008-12-05 | 2010-06-23 | 中国人民解放军军事医学科学院毒物药物研究所 | Thermal sensitive liposome preparation containing camptothecin antineoplastic agents |
-
2011
- 2011-06-09 CN CN2011101531428A patent/CN102210870B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1883454A (en) * | 2006-06-01 | 2006-12-27 | 复旦大学 | Liposome administration system with tumor necrosis targeted antibody for preposition and application thereof |
| CN101406455A (en) * | 2008-11-28 | 2009-04-15 | 南京中医药大学 | Strychnine composite phospholipid liposome, and preparation method thereof and use in pharmacy |
| CN101744767A (en) * | 2008-12-05 | 2010-06-23 | 中国人民解放军军事医学科学院毒物药物研究所 | Thermal sensitive liposome preparation containing camptothecin antineoplastic agents |
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102688153A (en) * | 2012-06-05 | 2012-09-26 | 东南大学 | Blank liposome prepared by compounding phospholipid and preparation method thereof |
| CN103404844A (en) * | 2013-04-26 | 2013-11-27 | 江南大学 | Composition capable of enhancing stability of lycopene and having function of relieving physical fatigue and preparation method thereof |
| CN104785178A (en) * | 2015-03-26 | 2015-07-22 | 江南大学 | Thermosensitive liposome as well as preparation method and application thereof |
| CN107998070A (en) * | 2017-12-12 | 2018-05-08 | 南京中医药大学 | A kind of composite phospholipid carrier and its application in percutaneous drug administration preparation is prepared |
| CN108030929A (en) * | 2017-12-19 | 2018-05-15 | 浙江农林大学 | A kind of preparation method of Lazer's property liposome comprising berberine |
| CN108030929B (en) * | 2017-12-19 | 2021-02-09 | 浙江农林大学 | Preparation method of double-sensitive liposome containing berberine |
| CN108669213A (en) * | 2018-04-26 | 2018-10-19 | 天津大学 | A kind of nisin thermal sensitive liposomes and its preparation method and application |
| CN108669213B (en) * | 2018-04-26 | 2021-06-08 | 天津大学 | Nisin thermosensitive liposome and preparation method and application thereof |
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