CN106166140A - A kind of palmitoyl ascorbate liposome - Google Patents

A kind of palmitoyl ascorbate liposome Download PDF

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CN106166140A
CN106166140A CN201610294387.5A CN201610294387A CN106166140A CN 106166140 A CN106166140 A CN 106166140A CN 201610294387 A CN201610294387 A CN 201610294387A CN 106166140 A CN106166140 A CN 106166140A
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acid
liposome
palmitoyl ascorbate
cholesterol
palmitoyl
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卢杨
王子厚
杨越
陈西敬
赵娣
李宁
陈琪
马恩龙
张玲
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    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
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    • A61K9/127—Liposomes
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K31/33—Heterocyclic compounds
    • A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/365—Lactones
    • A61K31/375—Ascorbic acid, i.e. vitamin C; Salts thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/28—Steroids, e.g. cholesterol, bile acids or glycyrrhetinic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner

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Abstract

The present invention relates to a kind of Liposomal formulation for drug administration by injection tumor, particularly relate to the preparation method of a kind of palmitoyl ascorbate liposome.The experiment in vivo and vitro of palmitoyl ascorbate all finds that it has notable antitumor drug effect, and toxicity is less, it is administered through modes such as vein, tremulous pulse, muscle, subcutaneous, abdominal cavities, especially for the insertion administration such as intravenous injection or intratumor injection, improves effectiveness and the safety of clinical practice.The preparation method technique that the present invention uses is simple, with low cost, and the liposomal particle size prepared is uniform, and envelop rate is high, and Release Performance is good, and extracorporeal suppression tumor cell growth and anti-tumor in vivo proliferative effect been significantly enhanced.

Description

A kind of palmitoyl ascorbate liposome
Technical field
The present invention relates to a kind of compositions Liposomal formulation for drug administration by injection tumor, particularly relate to one The preparation method of palmitoyl ascorbate liposome.
Background technology
Palmitoyl ascorbate (L-palmitoyl ascorbate, PA, Figure 14 be shown in chemical structural formula) is ascorbic acid A kind of fat-soluble derivant, due to the implantation of Palmic acid so that it is existing hydrophilic ascorbyl, have again the Petiolus Trachycarpi of oleophylic Acyl ester, thus become a kind of excellent antioxidant.Compared with ascorbic acid, its stability is higher, possesses more preferable physics and chemistry Character.After fat-soluble enhancing, palmityl ascorbic acid enters the ability of cell to be strengthened, and drug effect strengthens.Therefore itself and ascorbic acid Relatively, being administered through vein, tremulous pulse, muscle, subcutaneous, abdominal cavity etc., the particularly administering mode such as intratumor injection, insertion administration can be more Add and significantly strengthen Synergistic action and safety.And with ascorbic acid similarly, palmityl ascorbic acid can be selective Playing drug effect in tumor cell, little to normal cytotoxicity, therefore toxic and side effects is less.Additionally, hydrophobic characteristic makes Palmityl ascorbic acid is easier to by liposome entrapment, prepares the preferable liposome of slow release effect.
Antitumor drug, also can injuring normal cell medicine while playing antitumor cell effect.One for the treatment of Medicine orientation is key challenge is how to be transported to cancer cell and don't injuring normal cell.Liposome is targeting drug delivery system A kind of novel form, this system is current pharmaceutics drug-supplying system up-to-date, the most promising.It can be by drug selectivity ground It is transported to target spot position, plays therapeutical effect, the most do not affect the function of normal cell, tissue or organ simultaneously, thus reach to carry High curative effect, the purpose of minimizing toxic and side effects.
Report currently, with respect to palmitoyl ascorbate liposome is less.The Vladimir of Northeast USA university Professors P.Torchilin etc. report a kind of vitamin C palmityl ester liposome preparation, its palmitoyl ascorbate and phospholipid The ratio of cholesterol is 52.9: 100, and phospholipid and cholesterol ratio are 2: 1.It is contemplated that develop a kind of palmityl ascorbic acid The liposome of ester, to improve targeting, reduces toxic and side effects, improves curative effect.Meanwhile, preparation method of the present invention is suitable for Industry Promotion.
Summary of the invention
The solving the technical problem that of the present invention is to provide a kind of has good envelop rate, drug loading and stability；And And centralized particle diameter, size distribution uniform palmitoyl ascorbate liposome.
For solving the technical problem of the present invention, adopt the following technical scheme that
A kind of palmitoyl ascorbate liposome of the present invention, including following each component: palmitoyl ascorbate: phosphorus Lipid material: cholesterol analog.
And the weight part ratio between each component is as follows:
Palmitoyl ascorbate: phospholipid substance: cholesterol analog=0.06～60: 203.3～290: 96.7～ 9.67。
Preferably palmitoyl ascorbate: phospholipid substance: cholesterol analog=6～60: 203.3～273: 96.7 ～9.67.
Preferred palmitoyl ascorbate: phospholipid substance: cholesterol analog=6～24: 203.3～273: 96.7～27.
In compositions, each component is also selected from following weight portion:
The weight portion of ascorbyl palmitate is: 0.06～3；0.06～6；0.06～15；0.06～24；0.06～ 60；3～6；3～15；3～24；3～60；6～15；6～24；6～60；15～24；15～60；24～60；24；
The weight portion of phospholipid substance is: 203.3～225；203.3～250；203.3～273；203.3～290；225 ～250；225～273；225～290；250～275；250～290；275～290；
The weight portion of cholesterol analog is 96.7～75；96.7～50；96.7～27；96.7～9.67；75～50；75 ～27；75～9.67；50～27；50～9.67；27～9.67；
Preferably phospholipid substance is selected from phosphatidylcholine class, phosphatidyl glycerol class, phosphatidyl-4 alcohols, phosphatidyl ethanol Amine, sphingomyelin apoplexy due to endogenous wind at least one.
Preferably phospholipid substance is selected from strand or dichain phospholipids；
Preferably dichain phospholipids selected from DPPC (DPPC), tin dilaurate phosphatidylcholine (DLPC), Two myristic acid phosphatidylcholines (DMPC), distearoyl phosphatidylcholine (DSPC), two Semen Myristicae acid phosphatidyl glycerols, two Laurels Acid phosphatidyl glycerol, two palmitic acid phosphatidyl glycerols, DSPG, dilinoleic acid phosphatidylinositols, two Semen Myristicaes Acid phosphatidic acid, tin dilaurate phosphatidic acid, two palmitic acid phosphatidic acid, distearyl acid phosphatidic acid, two oleic acid Phosphatidylserine,
Preferably strand phospholipid selected from single Palmic acid phosphatidylcholine (MPPC), mono laurate phosphatidylcholine (MLPC), Single myristic acid phosphatidylcholine (MMPC), MSPC (MSPC), single Semen Myristicae acid phosphatidyl glycerol, single Laurel Acid phosphatidyl glycerol, single palmitic acid phosphatidyl glycerol, monostearate phosphatidyl glycerol, single Semen Myristicae acid phosphatidic acid, mono laurate phosphorus Fat acid, single palmitic acid phosphatidic acid, monostearate phosphatidic acid, single oleic acid Phosphatidylserine, single linoleic acid phosphatidylinositols,
Preferably phospholipid substance is further selected from soybean phospholipid, lecithin, cuorin, cephalin at least one.
Described cephalin can be naturally occurring or synthetic cephalin.
Preferred phospholipid substance is selected from two myristic acid phosphatidylcholines, DPPC, Semen sojae atricolor phosphorus In fat, lecithin, cephalin, cuorin at least one.
Most preferably phospholipid substance is selected from two myristic acid phosphatidylcholines, DPPC, Semen sojae atricolor phosphorus In fat at least one.
Preferred cholesterol analog is selected from cholesterol, Cholesteryl hemisuccinate, sitosterol at least one.
Most preferably cholesterol analog is selected from cholesterol.
In a preferred embodiment of the invention, the mass parts ratio (or weight ratio) between each component is permissible Selected from following scope combination in any:
Preferably ascorbyl palmitate: two myristic acid phosphatidylcholines: cholesterol=0.06～0.21: 6～24: 203.3～273: 96.7～27.
Preferably ascorbyl palmitate: two myristic acid phosphatidylcholines: cholesterol=0.12: 6～24: 203.3～ 273: 96.7～27.
Most preferably ascorbyl palmitate: two myristic acid phosphatidylcholines: cholesterol=0.12: 15～24: 203.3 ～273: 96.7～27.
In compositions, each component is also selected from following weight portion:
The weight portion of ascorbyl palmitate is: 6～15,6～24,15～24；
The weight portion of two myristic acid phosphatidylcholines is: 203.3～225；203.3～250；203.3～273；225～ 250；225～273；250～275；
The weight portion of cholesterol is 96.7～75；96.7～50；96.7～27；75～50；75～27；50～27；
In above-mentioned preferred embodiment, the mass parts ratio (or weight ratio) between most preferred each component is selected from:
Most preferably ascorbyl palmitate: two myristic acid phosphatidylcholines: cholesterol=0.12: 24: 203.3: 96.7。
Most preferably ascorbyl palmitate: two myristic acid phosphatidylcholines: cholesterol=0.12: 24: 250: 50.
Most preferably ascorbyl palmitate: two myristic acid phosphatidylcholines: cholesterol=0.12: 24: 273: 27.
In a preferred embodiment of the invention, the mass parts ratio (or weight ratio) between each component is permissible Selected from following scope combination in any:
Preferably ascorbyl palmitate: soybean phospholipid: sitosterol=0.06～0.21: 6～24: 203.3～273: 96.7～27.
More preferably ascorbyl palmitate: soybean phospholipid: sitosterol=0.12: 6～24: 203.3～273: 96.7～ 27。
Most preferably ascorbyl palmitate: soybean phospholipid: sitosterol=0.12: 15～24: 203.3～273: 96.7～ 27。
In compositions, each component is also selected from following weight portion:
The weight portion of ascorbyl palmitate is: 6～15,6～24,15～24；
The weight portion of soybean phospholipid is: 203.3～225；203.3～250；203.3～273；225～250；225～ 273；250～275；
The weight portion of sitosterol is 96.7～75；96.7～50；96.7～27；75～50；75～27；50～27；
In above-mentioned preferred embodiment, the mass parts ratio (or weight ratio) between most preferred each component is selected from:
Most preferably ascorbyl palmitate: soybean phospholipid: sitosterol=0.12: 24: 203.3: 96.7.
Most preferably ascorbyl palmitate: soybean phospholipid: sitosterol=0.12: 24: 250: 50.
Most preferably ascorbyl palmitate: soybean phospholipid: sitosterol=0.12: 24: 273: 27.
In a preferred embodiment of the invention, the mass parts ratio (or weight ratio) between each component is permissible Selected from following scope combination in any:
Preferably ascorbyl palmitate: DPPC: Cholesteryl hemisuccinate=0.06～ 0.12: 6～24: 203.3～273: 96.7～27.
More preferably ascorbyl palmitate: DPPC: Cholesteryl hemisuccinate=0.12: 6～ 24: 203.3～273: 96.7～27.
Most preferably ascorbyl palmitate: DPPC: Cholesteryl hemisuccinate=0.12: 15～ 24: 203.3～273: 96.7～27.
In compositions, each component is also selected from following weight portion:
The weight portion of ascorbyl palmitate is: 6～15,6～24,15～24；
The weight portion of DPPC is: 203.3～225；203.3～250；203.3～273；225～ 250；225～273；250～275；
The weight portion of Cholesteryl hemisuccinate is 96.7～75；96.7～50；96.7～27；75～50；75～27；50 ～27；
In above-mentioned preferred embodiment, the mass parts ratio (or weight ratio) between most preferred each component is selected from:
Most preferably ascorbyl palmitate: DPPC: Cholesteryl hemisuccinate=0.12: 24: 203.3∶96.7。
Most preferably ascorbyl palmitate: DPPC: Cholesteryl hemisuccinate=0.12: 24: 250∶50。
Most preferably ascorbyl palmitate: DPPC: Cholesteryl hemisuccinate=0.12: 24: 273∶27。
In a preferred embodiment of the invention, the mass parts ratio (or weight ratio) between each component is permissible Selected from following scope combination in any:
Preferably ascorbyl palmitate: soybean phospholipid: cholesterol=0.06～0.21: 6～24: 203.3～273: 96.7～27.
More preferably ascorbyl palmitate: soybean phospholipid: cholesterol=0.12: 6～24: 203.3～273: 96.7～ 27。
Most preferably ascorbyl palmitate: soybean phospholipid: cholesterol=0.12: 15～24: 203.3～273: 96.7～ 27。
In compositions, each component is also selected from following weight portion:
The weight portion of ascorbyl palmitate is: 6～15,6～24,15～24；
The weight portion of soybean phospholipid is: 203.3～225；203.3～250；203.3～273；225～250；225～ 273；250～275；
The weight portion of cholesterol is 96.7～75；96.7～50；96.7～27；75～50；75～27；50～27；
In above-mentioned preferred embodiment, the mass parts ratio (or weight ratio) between most preferred each component is selected from:
Most preferably ascorbyl palmitate: soybean phospholipid: cholesterol=0.12: 24: 203.3: 96.7.
Most preferably ascorbyl palmitate: soybean phospholipid: cholesterol=0.12: 24: 250: 50.
Most preferably ascorbyl palmitate: soybean phospholipid: cholesterol=0.12: 24: 273: 27.
Palmitoyl ascorbate composite lipidosome prepared by the present invention, particle diameter is 80-200nm, and envelop rate is more than 70%, The drug loading of palmitoyl ascorbate is 0.1%～40%.
Preferably prescription is: particle diameter: 100～200nm, envelop rate: ＞ 85%, the drug loading of palmitoyl ascorbate: 1%～20%.
Second aspect present invention provides the preparation method of a kind of above-mentioned palmitoyl ascorbate liposome, the method bag Include following steps:
A, the palmitoyl ascorbate of recipe quantity, phospholipid substance, cholesterol analog are dissolved in organic solvent；
B, evaporating organic solvent, prepare uniform lipid membrane in bottle wall；
C, be dried；
D, add distilled water, use and ultrasonic lipid membrane is washed down；
Probe Ultrasonic Searching under E, low temperature, to obtain final product.
In step A, preferred organic solvent can be dichloromethane, chloroform, acetone, ethanol, ether, oxolane. Preferred solvent is selected from dichloromethane, chloroform, acetone；Most preferably organic solvent is selected from chloroform.
In step B, evaporating organic solvent preferably employs the mode of rotating pressure-decreasing；When preferably rotating pressure-decreasing evaporates Temperature is selected from 30 DEG C～80 DEG C；Preferably temperature is selected from 50 DEG C～75 DEG C；Preferred temperature is selected from 65 DEG C～75 DEG C；Most preferably Temperature be 70 DEG C；
In step C, the mode being dried: preferably employ vacuum drying；The temperature being dried is set as 30 DEG C～90 DEG C；Temperature is selected From 40 DEG C～90 DEG C；Preferably temperature is selected from 50 DEG C～85 DEG C；Preferred temperature is selected from 70 DEG C～80 DEG C；Most preferably temperature It it is 75 DEG C；
In step E, Probe Ultrasonic Searching intensity and duration are set as 30%～90%, 10s～50s, and ultrasonic 1～7s stops 1s, follows Ring number of times is 1～8 time.
Third aspect present invention provides the application in preparing antitumor drug of the palmitoyl ascorbate liposome.
Described tumor selected from melanoma, gastric cancer, pulmonary carcinoma, breast carcinoma, renal carcinoma, hepatocarcinoma, oral cavity epidermal carcinoma, cervical cancer, Ovarian cancer, cancer of pancreas, carcinoma of prostate, colon cancer, bladder cancer, cerebroma, the esophageal carcinoma.
Advantageous Effects
The palmitoyl ascorbate liposome of the present invention has good envelop rate, drug loading and stability.
The palmitoyl ascorbate liposome toxicity of the present invention reduces, and safety is high.Improve the safety of clinical application Property and effectiveness, have important actual application prospect.
The preparation method of the palmitoyl ascorbate liposome of the present invention has that simple process, preparation cost be low, technique Favorable reproducibility, experiment prescription and technique are scalable to the features such as production application, thus the application to industrialization has directive significance.
Accompanying drawing explanation
Fig. 1. palmitoyl ascorbate pharmacy in vitro experiment in MCF-7 cell.
Fig. 2. palmitoyl ascorbate pharmacy in vitro experiment in HepG-II cell.
Fig. 3. palmitoyl ascorbate pharmacy in vitro experiment in A549 cell.
Fig. 4. palmitoyl ascorbate pharmacy in vitro experiment in SGC-7901 cell.
Fig. 5. palmitoyl ascorbate pharmacy in vitro experiment in BxPC-3 cell.
Fig. 6. palmitoyl ascorbate pharmacy in vitro experiment in Hela cell.
Fig. 7. palmitoyl ascorbate pharmacy in vitro experiment in U251 cell.
Fig. 8. palmitoyl ascorbate pharmacy in vitro experiment in PC-3 cell.
Fig. 9. palmitoyl ascorbate pharmacy in vitro experiment in LOVO cell.
Figure 10. palmitoyl ascorbate pharmacy in vitro experiment in HELF cell.
Figure 11 .PA pharmacy in vitro experiment in U87 cell.
Figure 12 .PA pharmacy in vitro experiment in HCT-116 cell.
Figure 13. ascorbyl palmitate solution group and liposome group blood in rat body after the administration of intravenous 30mg/kg Concentration-time graph.
Figure 14. the chemical structural formula of palmitoyl ascorbate
Detailed description of the invention
Embodiment 1: a kind of palmitoyl ascorbate liposome formula and preparation thereof are as follows:
Experimental technique: described palmitoyl ascorbate liposome preparation technology is: by palmitoyl ascorbate, two Myristic acid phosphatidylcholine, cholesterol are dissolved in chloroform, and vortex adds 500mL eggplant type flask after being completely dissolved In, 70 DEG C of rotating pressure-decreasing evaporating organic solvent, in bottle wall, prepare uniform lipid membrane, be placed in vacuum drying oven 75 After being dried overnight at DEG C, add distilled water, ultrasonic lipid membrane is washed lower after, Probe Ultrasonic Searching 2 circulation under ice-water bath (30%, 20s, ultrasonic 1s stop 1s), to obtain final product.
Liposome evaluation methodology:
The mensuration of the particle diameter of liposome: use laser granulometry to measure.
The liposome of envelop rate and the mensuration of drug loading: 0.3mL crosses sephadex column, is eluting with normal saline Liquid, after the most free medicine and liposome, collects liposome components, is measured with HPLC after methanol breakdown of emulsion constant volume, meter Calculate to obtain concentration of liposomes C1.Separately take 0.3mL liposome methanol constant volume to upper prop after liposome components same volume, then enter Sample also calculates total concentration C after liposome breakdown of emulsion0.According to below equation computational envelope rate (Encapsulation Efficiency, EE%) and drug loading (Drug Loading Efficiency, DLE%):
E E % = C 1 C 0 × 100 %
Experimental result:
The experimental result of table 1. embodiment 1
Palmitoyl ascorbate: PA, two myristic acid phosphatidylcholine: DMPC, cholesterol: CHOL
Preparation evaluation standard: particle diameter: 100～200nm, envelop rate: ＞ 85%, drug loading: 1%～20% is qualified preparation
Embodiment 2: a kind of palmitoyl ascorbate liposome formula and preparation thereof are as follows:
Experimental technique: described palmitoyl ascorbate liposome preparation technology is: by palmitoyl ascorbate, big Fabaceous lecithin, sitosterol are dissolved in chloroform, and vortex adds in 500mL eggplant type flask after being completely dissolved, and 60 DEG C of rotations subtract Pressure evaporating organic solvent, prepares uniform lipid membrane in bottle wall, is placed in vacuum drying oven and is dried overnight at 35 DEG C After, add distilled water, ultrasonic lipid membrane is washed lower after, under ice-water bath, (80%, 50s, ultrasonic 1s stops in Probe Ultrasonic Searching 4 circulation 1s), to obtain final product.
Experimental result:
The experimental result of table 2. embodiment 2
Palmitoyl ascorbate: PA
Preparation evaluation standard: particle diameter: 100～200nm, envelop rate: ＞ 85%, drug loading: 1%～20% is qualified preparation
Embodiment 3: a kind of palmitoyl ascorbate liposome formula and preparation thereof are as follows:
Experimental technique: described palmitoyl ascorbate liposome preparation technology is: by palmitoyl ascorbate, two Palmic acid phosphatidylcholine, Cholesteryl hemisuccinate are dissolved in chloroform, and vortex adds 500mL eggplant after being completely dissolved In shape flask, 70 DEG C of rotating pressure-decreasing evaporating organic solvent, in bottle wall, prepare uniform lipid membrane, be placed in vacuum drying After case is dried overnight at 75 DEG C, add distilled water, ultrasonic lipid membrane is washed lower after, Probe Ultrasonic Searching 6 circulation under ice-water bath (30%, 60s, ultrasonic 1s stops 1s), to obtain final product.
Experimental result:
The experimental result of table 3. embodiment 3
Palmitoyl ascorbate: PA, DPPC: DMPC, Cholesteryl hemisuccinate: CHOL
Preparation evaluation standard: particle diameter: 100～200nm, envelop rate: ＞ 85%, drug loading: 1%～20% is qualified preparation
Embodiment 4: a kind of palmitoyl ascorbate liposome formula and preparation thereof are as follows:
Experimental technique: described palmitoyl ascorbate liposome preparation technology is: by palmitoyl ascorbate, big Fabaceous lecithin, cholesterol are dissolved in chloroform, and vortex adds in 500mL eggplant type flask after being completely dissolved, and 70 DEG C of rotations subtract Pressure evaporating organic solvent, prepares uniform lipid membrane in bottle wall, is placed in vacuum drying oven and is dried overnight at 65 DEG C After, add distilled water, ultrasonic lipid membrane is washed lower after, under ice-water bath, (50%, 60s, ultrasonic 1s stops in Probe Ultrasonic Searching 2 circulation 1s), to obtain final product.
Experimental result:
The experimental result of table 4. embodiment 4
Palmitoyl ascorbate: PA, cholesterol: CHOL
Preparation evaluation standard: particle diameter: 100～200nm, envelop rate: ＞ 85%, drug loading: 1%～20% is qualified preparation
The preparation of existing document liposome is also compared with this patent liposome
Experiment 1
The palmitoyl ascorbate of Vladimir professor P.Torchilin with reference to Northeast USA university et al. report Liposomal formulation document, prepares liposome by the prescription proportioning of report in the document, and measures its envelop rate, particle diameter.
In the document, phosphatidase 2 00mg, cholesterol 100mg, the system that palmitoyl ascorbate 150mg is related to by this patent Preparation Method is prepared.
Experimental result: in preparation process, is adding distilled water, ultrasonic when washing lipid membrane, has obvious granule to separate out, it is impossible to Continue preparation.
Pharmacological evaluation
Experimental example 1: the pharmacy in vitro experiment of palmitoyl ascorbate
Mtt assay measures activity of tumor cells
The take the logarithm cell strain of trophophase, after 0.25% trypsinization, is cultivated with the RPMI1640 containing 10% hyclone Liquid is made into single cell suspension, with 3 × 103The density of individual/mL is inoculated in 96 orifice plates, and every hole adds 100 μ L cell suspension, is placed in 37 DEG C, 5%CO2Incubator is cultivated in aseptic culture case 24h, after cell is the most adherent, removes original culture fluid, every hole The variable concentrations palmitoyl ascorbate 100 μ L that addition configures with culture fluid.Matched group adds the cultivation containing 0.1%DMSO Liquid, cultivates 48h for 37 DEG C.Cultivation is inhaled after terminating and is abandoned medicinal liquid, and every hole adds PBS 100 μ L and washes away the medicinal liquid of residual, and every hole adds and contains 10%5mg/mL MTT working solution culture fluid 100 μ L, 37 DEG C are continued to cultivate 4h.Supernatant is abandoned in suction, and every hole adds 150 μ L DMSO, 10min is with abundant dissolving crystallized thing in vibration, measures OD value under microplate reader 570nm, calculates after giving medicine swollen by following equation The survival rate of oncocyte, and use SPSS data processing software calculation of half inhibitory concentration (IC50)。
According to the suppression ratio of each acute drug, apply SPSS statistical analysis software calculation of half inhibitory concentration IC50.
Above-mentioned cell strain is selected from: MCF-7 cell strainHJ2mm, human hepatoma cell strain HepG-II, human lung carcinoma cell line A549, human stomach cancer cell line SGC-7901, people pancreas adenocarcinoma cell strain BxPC-3, human cervical carcinoma cell lines Hela, people's glue in situ Matter tumor cell strain U251, human prostate cancer cell line PC-3, human colon cancer cell strain LOVO, human lung cancer cell A549 strain HELF, Human glioma cell U-87, human colon cancer cell HCT-116.
The exercising result of MCF-7 cell strainHJ2mm is shown in Fig. 1 by PA.
The exercising result of human hepatoma cell strain HepG-II is shown in Fig. 2 by PA.
The exercising result of human lung carcinoma cell line A549 is shown in Fig. 3 by PA.
The exercising result of human stomach cancer cell line SGC-7901 is shown in Fig. 4 by PA.
The exercising result of people pancreas adenocarcinoma cell strain BxPC-3 in situ is shown in Fig. 5 by PA.
The exercising result of human cervical carcinoma cell lines Hela is shown in Fig. 6 by PA.
The exercising result of human glioma cells strain U251 is shown in Fig. 7 by PA.
The exercising result of human prostate cancer cell line PC-3 is shown in Fig. 8 by PA.
The exercising result of human colon cancer cell strain LOVO is shown in Fig. 9 by PA.
The exercising result of human lung cancer cell A549 strain HELF is shown in Figure 10 by PA.
The exercising result of human glioma cell U-87 is shown in Figure 11 by PA.
The exercising result of human colon cancer cell HCT-116 is shown in Figure 12 by PA.
Table 1. palmitoyl ascorbate IC in different cell strains50Value
Pharmacokinetic experiments
Experimental example 1: palmitoyl ascorbate liposome compares with the pharmacokinetics of palmitoyl ascorbate solution
Taking rat 6, be randomly divided into two groups by body weight, often three rats of group, give 6mg/mL by 30mg/kg dosage intravenous (rat is compiled for palmitoyl ascorbate solution (rat numbered No.1, No.2, No.3), palmitoyl ascorbate liposome Number be No.4, No.5, No.6), after being administered precontract 0.25h and being administered 0.083h, 0.25h, 0.5h, 1h, 1.5h, 2h, 4h, 8h, 12h, 24h take blood, in about 200 μ L to heparinised tubes, are centrifuged 5min in 8000rpm, isolate blood plasma, in-80 DEG C Preserve for test.
Table 2. rat intravenous gives the blood drug level (μ g/L) after 30mg/kg palmitoyl ascorbate solution
Table 3. rat intravenous gives the pharmacokinetic parameter after 30mg/kg palmitoyl ascorbate solution in rat body
Table 4. rat intravenous gives the blood drug level (μ g/L) after 30mg/kg palmitoyl ascorbate liposome
The pharmacokinetics ginseng that table 5 rat intravenous gives after 30mg/kg palmitoyl ascorbate liposome in rat body Number
Experimental result: see Figure 13 and Biao 2-5.Medicine moves parameter and shows, intravenous gives rat same dose of ascorbic acid palm fibre After palmitic acid acid esters solution and liposome, the half-life of liposome group is 4 times of solution group, shows that liposome slow down drug release, Extend the action time of medicine, enhance drug effect.

Claims (12)

1. a palmitoyl ascorbate liposome, it is characterised in that include following each component: palmitic acid acyl acid ascorbyl ester, phosphorus Lipid material and cholesterol analog；And the weight part ratio between each component is as follows:
Palmitoyl ascorbate: phospholipid substance: cholesterol analog=0.06～60: 203.3～290: 96.7～9.67.
Palmitoyl ascorbate liposome the most according to claim 1, it is characterised in that palmitoyl ascorbate: phospholipid Class material: cholesterol analog=0.06～0.12: 6～24: 203.3～273: 96.7～27.
3. according to the palmitoyl ascorbate liposome any one of claim 1 or 2, it is characterised in that described phosphorus Lipid material is selected from phosphatidylcholine class, phosphatidyl glycerol class, phosphatidyl-4 alcohols, cytoskeletal protein, sphingomyelin Apoplexy due to endogenous wind at least one；
Described cholesterol analog is selected from cholesterol, Cholesteryl hemisuccinate, sitosterol at least one.
Palmitoyl ascorbate liposome the most according to claim 3, described phospholipid substance is selected from strand or double-strand phosphorus Fat.
Palmitoyl ascorbate liposome the most according to claim 4, described dichain phospholipids is selected from two Palmic acid phosphatidyls Choline (DPPC), tin dilaurate phosphatidylcholine (DLPC), two myristic acid phosphatidylcholines (DMPC), distearoylphosphatidyl The acid of choline (DSPC), two Semen Myristicae acid phosphatidyl glycerols, tin dilaurate phosphatidyl glycerol, two palmitic acid phosphatidyl glycerols, distearyl Phosphatidyl glycerol, dilinoleic acid phosphatidylinositols, two Semen Myristicae acid phosphatidic acid, tin dilaurate phosphatidic acid, two palmitic acid phosphatidic acid, two Stearic acid phosphatidic acid, two oleic acid Phosphatidylserine；
Described strand phospholipid is selected from single Palmic acid phosphatidylcholine (MPPC), mono laurate phosphatidylcholine (MLPC), single meat Myristic acid phosphatidylcholine (MMPC), MSPC (MSPC), single Semen Myristicae acid phosphatidyl glycerol, mono laurate phosphorus Phosphatidyl glycerol, single palmitic acid phosphatidyl glycerol, monostearate phosphatidyl glycerol, single Semen Myristicae acid phosphatidic acid, mono laurate phosphatidic acid, Single palmitic acid phosphatidic acid, monostearate phosphatidic acid, single oleic acid Phosphatidylserine, single linoleic acid phosphatidylinositols.
Palmitoyl ascorbate liposome the most according to claim 3, described phospholipid substance is selected from two myristic acid phosphorus In phosphatidylcholine, DPPC, soybean phospholipid, lecithin, cephalin, cuorin at least one；
Described cholesterol analog is selected from cholesterol.
Palmitoyl ascorbate liposome the most according to claim 3, it is characterised in that include following each component: Vitamin C Acid cetylate, two myristic acid phosphatidylcholine and cholesterol；And the weight part ratio between each component is as follows:
Ascorbyl palmitate: two myristic acid phosphatidylcholines: cholesterol=6～24: 203.3～273: 96.7～27.
Palmitoyl ascorbate liposome the most according to claim 3, it is characterised in that include following each component cetylate : soybean phospholipid: sitosterol=: 6～24: 203.3～273: 96.7～27.
Palmitoyl ascorbate liposome the most according to claim 3, it is characterised in that include following each component: Vitamin C Acid cetylate: DPPC: Cholesteryl hemisuccinate；And the weight part ratio between each component is as follows:
Ascorbyl palmitate: DPPC: Cholesteryl hemisuccinate=6～24: 203.3～273: 96.7～27.
Palmitoyl ascorbate liposome the most according to claim 3, it is characterised in that include following each component: Vitamin C Acid cetylate, soybean phospholipid and cholesterol；And the weight part ratio between each component is as follows:
Ascorbyl palmitate: soybean phospholipid: cholesterol=6～24: 203.3～273: 96.7～27.
The application in preparing antitumor drug of the palmitoyl ascorbate liposome of 11. any one of claim 1-10.
12. application according to claim 11, it is characterised in that: described tumor is selected from melanoma, gastric cancer, pulmonary carcinoma, mammary gland Cancer, renal carcinoma, hepatocarcinoma, oral cavity epidermal carcinoma, cervical cancer, ovarian cancer, cancer of pancreas, carcinoma of prostate, colon cancer, bladder cancer, cerebroma, esophagus Cancer.
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114081963A (en) * 2021-11-16 2022-02-25 上海理工大学 Nano-carrier for improving bioavailability of active peptide and preparation and application thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101507708A (en) * 2008-03-28 2009-08-19 中国人民解放军济南军区第四○一医院 Preparation method of liposome
US20150057341A1 (en) * 2013-08-22 2015-02-26 Stephen C. Perry Composition with Pharmacological Effect for Vaporizing and Method

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101507708A (en) * 2008-03-28 2009-08-19 中国人民解放军济南军区第四○一医院 Preparation method of liposome
US20150057341A1 (en) * 2013-08-22 2015-02-26 Stephen C. Perry Composition with Pharmacological Effect for Vaporizing and Method

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
KRISTL ET AL.: "Effect of colloidal carriers on ascorbyl palmitate stability", 《EUROPEAN JOURNAL OF PHARMACEUTICAL SCIENCES》 *
SAWANT ET AL.: "Palmitoyl Ascorbate Liposomes and Free Ascorbic Acid: Comparison of Anticancer Therapeutic Effects Upon Parenteral Administration", 《PHARM RES》 *
SAWANT ET AL.: "Palmitoyl Ascorbate-Loaded Polymeric Micelles: Cancer Cell Targeting and Cytotoxicity", 《PHARM RES》 *
孟胜男等主编: "《药剂学》", 31 January 2016, 中国医药科技出版社 *
曾戎等编著: "《生物医用仿生高分子材料》", 31 October 2010, 华南理工大学出版社 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114081963A (en) * 2021-11-16 2022-02-25 上海理工大学 Nano-carrier for improving bioavailability of active peptide and preparation and application thereof
CN114081963B (en) * 2021-11-16 2023-09-26 上海理工大学 Nanometer carrier for improving bioavailability of active peptide and preparation and application thereof

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