CN102475682B - Berberine liposome and preparation method thereof - Google Patents
Berberine liposome and preparation method thereof Download PDFInfo
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Abstract
The invention belongs to the technical field of medicines, relating to a berberine liposome and a preparation method thereof. The invention also relates to an application of the berberine liposome. The berberine liposome comprises a membrane material and berberine, wherein the membrane material is a phospholipid bilayer and is composed of phospholipids, cholesterol and polyethylene glycol lipid derivates, granularity of the liposome is 30-200nm, and encapsulation efficiency is more than 80%. Targeted substance lipid derivates also can be added into the membrane material, such as a targeted antibody, a ligand and the like, comprising transferrin, lactoferrin, folic acid, galactose, glucose, RGD (arginine-glycine-aspartic acid) and albumin and also comprising a cell penetrating peptide lipid derivate. In the invention, encapsulation efficiencies of liposomes formed by different membrane materials are all higher than 80% and even reach up to 100%, and the process is stable.
Description
Technical field
The invention belongs to medical technical field, relate to berberine liposome and preparation method thereof.The invention still further relates to the purposes of berberine liposome.
Background technology
Berberine belongs to isoquinoline alkaloid, Recent study is found, 200810113680.2), the many-sides such as hyperlipidemia, inflammation, antibacterial and viral infection, cerebral ischemic injury, mental sickness, Alzheimer (Alzheimer disease), osteoporosis have pharmacological action (berberine has good therapeutical effect to cardiovascular disease, application number: in treatment tumor, diabetes, cardiovascular disease for berberine and derivant thereof.
Berberine can suppress the activity of Topo I and Topo II simultaneously, i.e. economic benefits and social benefits inhibitor effectively disturbs DNA of tumor cell to copy and brings into play antitumous effect [Pharm Pharmacol, 2001,53:763-768]; Directly the double-spiral structure of the intercalation of DNA destroys the dna double chain, the DNA of inhibition tumor cell and protein synthesis [J Pharm Pharmacol, 2006,58:263-270.]; Can be by reducing the expression of cell periodic protein B 1 and increase Wee1, stopping of G2 in the inducing cell growth cycle/M stage makes tumor cell stop at G1 phase and G2 phase (Anticancer Res, 2006,26:1097-1104.); Mouse melanin tumor cell there is inhibitory action (J Pharmacol Exp Ther, 2007,323:636-649.), by reducing in the cell Ca2+control of the concentration COX-2 mrna expression and COX-2 to arachidonic catalytic activity, thereby suppress the generation of PGH2, effect (Chinese Pharmacological circular, 2005, the 21:950-953. of performance inhibition tumor cell propagation; J Ethnopharmacol, 1999,66:227-233.); Berberine can be by suppressing the activity of activator protein-1 (AP-1), the relevant signal path of blocking-up neoplasm metastasis; Also can reduce VEGF by proteolysis and lysine acetylation approach, and can directly suppress the epithelial formation of umbilical vein and movement.Also can the transfer of inhibition tumor cell by the generation that reduces UPA and matrix metalloproteinase.Can improve the dyscrasia disease that too much causes because of the L-6 synthesis secretion by the expression that suppresses L-6 mRNA, the resistance of enhancing body is from the ability of integral body raising body anti metastasis.Also can be by suppressing sick cell Bcl22(proto-oncogene), the unconventionality expression of mtP53 gene, promote the expression of wt P53 gene, apoptosis be tending towards normally, stop, the reverse precancerous lesion [Chinese combination of Chinese and Western medicine taste magazine), 2005,13:81-84.].
2O 30 or 40 years in century, it was a kind of special knot chair of end of chromosome, can prevent chromosomal DNA degraded, terminal fusion, disappearance and improper restructuring for the concept that proposes telomere, and it is stable to keep chromosomal complete sum.Recent study is found the telomere dystropy of human malignancies, and telomerase is high expressed in most of malignant cells, has become a novel targets of oncotherapy.There is research to find that berberine (45 μ molL-1) can reduce telomerase activation, induce the HL260 apoptosis [Int J Cancer, 1999,81:923-929]; Suppress nasopharyngeal carcinoma CNE-2 Cell Telomerase Activity, may improve curative effect as the auxiliary treatment of nasopharyngeal carcinoma. (Luo Biao. etc., berberine affects Shaanxi medical journal 2,007 36 (10) to human nasopharyngeal carcinoma CNE-2 Cell Telomerase Activity).
For absorption problem and liver metabolism loss problem in the body that solves berberine, improve its therapeutic effect, patent is arranged, and (ammonium sulphate gradient-film evaporation method prepares the method application number of berberine hydrochloride long circulating liposomes: 200710025536.9 applyings date: 2007-08-03) adopt ammonium sulphate gradient-film evaporation method that it is prepared into long circulating liposomes, concrete operation method is as follows: 1) cholesterol of Ovum Gallus domesticus Flavus lecithin, 1/3 Ovum Gallus domesticus Flavus lecithin quality and the Macrogol 2000 of 1/20-1/5 Ovum Gallus domesticus Flavus lecithin quality are dissolved in the ethanol; 2) decompression recycling ethanol forms film at chamber wall; Inject ammonium sulfate and make the dissolving of film material; Aquation 1H～2H; 3) take distilled water as dialysis medium dialysis 15～20H, make ammonium sulphate gradient, get blank liposome; 4) in blank liposome, add berberine hydrochloride solution; 5) 30 ℃～60 ℃ hatchings 10～60 minutes; 6) get solution after the hatching, the 12H that dialyses under different temperatures take distilled water as extracellular fluid dialysis is to outer liquid water white transparency; Dialysis solution shifts, standardize solution, and get final product, but its envelop rate of liposome of formulation and technology preparation is less than 40%.Another patent has been described the preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method method, and (the method application number of preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method: 200810200546.6 applyings date: 2008-09-26), problem is that envelop rate is also very low.The prepared berberine hydrochloride liposome particle size range of the reports such as Xu Lijun is 2. 4～4. 6 um, and envelop rate is preparation method and 2004 years 06 phase of quality research Journal of Chinese Hospital Pharmacy of 39. 77%(berberine hydrochloride liposomes); Zhang Fen etc. are take cholesterol and lecithin as the film material, film evaporation method prepare berberine hydrochloride liposome (the Study on Preparation Nanjing Normal University journal (natural science edition) 2006 of berberine hydrochloride liposome, 29(1): 56～58); Tan Lirong etc. adopt the preferred berberine hydrochloride liposome of uniform design, prepared liposome particle size range is 2.2 μ m～3.8 μ m (practical medical technologies magazine 11 phases in 2007 of the prescription of the preferred berberine hydrochloride liposome of uniform design), is not suitable for drug administration by injection.
Summary of the invention
Technical problem solved by the invention provides a kind of berberine liposome and its production and use.
In the berberine liposome preparation of the present invention,, during forming, its prescription contains the PEG lipid derivate, and envelop rate is greater than 80%, and particle diameter is less than 200nm.
Classical pH gradient is with citrate buffer solution aquation lipid film material, and then the preparation liposome adds alkaline matter in liposome turbid liquor, regulate outer aqueous pH values and carry out medicine carrying.As the product Evacet (trade name Myocet) that goes on the market, carrying out clinical liposomal vincristine body (Marqibo) etc., wherein Myocet is comprised of with " amycin (50mg) " " blank liposome (EPC 142.6 mg/ml, CH57.4 mg/ml, citrate buffer solution 57.6mg/ml) ", " sodium carbonate liquor (54.6 mg/ml, 3.1 ml) ", and namely citrate buffer solution is that aquation medium, sodium carbonate are pH adjusting agent; And the pH adjusting agent of Marqibo is sodium radio-phosphate,P-32 solution.
If the pH gradient with classics is loaded berberine, although envelop rate can reach more than 70%, but data are unstable, and formed and particle diameter by the liposome membrane material, the impacts such as hybrid mode (Preparation of berberine hydrochloride liposomes by active loading method Chinese Pharmaceutical Journal 01 phase in 2004), when spent ion exchange resin (fiber), ultrafiltration, dialysis, the methods such as molecular sieve are removed the citrate buffer solution of the outer water of blank liposome, simultaneously in conjunction with adding alkaline matter, such as sodium carbonate, sodium bicarbonate, sodium phosphate, regulate pH to 6.5～9.5, namely adopt citric acid gradient and pH gradient integrated processes to load berberine, can obtain the envelop rate greater than 80%, even reach 100% envelop rate, and technique favorable reproducibility, preparation stabilization, be not subjected to liposome membrane material and liposome grain diameter influence, and the medicine carrying time also can foreshorten to 5～10min.In addition, the gap between the normal blood vessels endotheliocyte is about 2nm usually, and tumor is because Fast Growth, and its vascular space can reach 100-780nm, and this passs medicine for targeting and lays a good foundation.Our experiment is found, adds PEG-DSPE in berberine liposome, and granularity is controlled at less than 200nm, has good antitumous effect.
Generally speaking; liposome is by phospholipid; cholesterol is the film material; wherein phospholipid can be selected from following at least a: natural phospholipid; semi-synthetic phospholipid; synthetic phospholipid; such as soybean lecithin; Ovum Gallus domesticus Flavus lecithin; phosphatidyl glycerol; EPG; phosphatidic acid; cardiolipin; sphingomyelins; the phosphatidic acid serine; phosphatidylinositols; PHOSPHATIDYL ETHANOLAMINE; hydrogenated soy phosphatidyl choline; hydrogenated yolk lecithin; distearoyl phosphatidylcholine; dipalmitoyl phosphatidyl choline; DOPC; dimyristoyl phosphatidyl choline; DLPC; DDPC; two decoyl phosphatidylcholines; DHPC; DSPG and salt thereof; DPPG and salt thereof; L-α-GLYCEROL,DIMYRISTOYL PHOSPHATIDYL and salt thereof; PE; two caprinoyl phosphatidyl glycerols; two decoyl phosphatidyl glycerols; two hexanoyl phosphatidyl glycerols; DSPE; DPPE; DOPE, DMPEA; two lauroyl PHOSPHATIDYL ETHANOLAMINE; two DSPGs and salt thereof; two DPPGs and salt thereof; two GLYCEROL,DIMYRISTOYL PHOSPHATIDYLs and salt thereof; two PEs; the distearyl phosphatidylinositols; two palmityl phosphatidylinositols; dioleoyl phospholipid acyl inositol; two myristoyl phosphatidylinositols; two lauroyl phosphatidylinositols; POPC; the inferior oleoyl phosphatidylcholine of palmityl; the inferior oleoyl phosphatidylcholine of stearoyl; stearoyl oleoyl phosphatidylcholine; stearoyl arachidonic phosphatidyl choline.The present invention once comparatively systematically studied/investigated different membrane materials and formed, although conventional liposome, as the conventional liposome that adopts soybean lecithin and cholesterol to form, envelop rate also can access more than 80%, even this is that existing all berberine liposome formulation and technology institutes are inaccessiable to reach 100%(), but the antineoplaston application facet has problems in vivo, i.e. the short problem of circulation time.Based on this, we add hydroaropic substance in the liposome membrane material, particularly HLB is greater than 8 surfactant, as use the hydrophilic polymer lipid material, comprise poly-DSPE-PEG(ethylene glycol-DSPE), DPPE-PEG(ethylene glycol-DPPE), DMPE-PEG(ethylene glycol-DMPEA), DLPE-PEG(ethylene glycol-two lauroyl PHOSPHATIDYL ETHANOLAMINE), DSPG-PEG(Polyethylene Glycol-DSPG), the CHOL-PEG(PEG-CHOL), DSPE-PVP(polyvidone-DSPE), DSPG-PVP(polyvidone-DSPG), CHOL-PVP(polyvidone-cholesterol), preferred ethylene glycol-two phosphatidyl ethanolamine derivant wherein; The molecular weight of PEG is 100～100000, preferred 1000～5000.In order to increase the targeting of liposome, still can add specific antibodies or aglucon lipid derivate in the liposome membrane material, such as various monoclonal antibodies, transferrins part, lactoferrin part, folic acid part, galactose part, glucose part, RGD, albumin, still comprise cell membrane penetrating peptide lipid derivate.
The used blank liposome of the present invention can by multiple appropriate method preparation, comprise such as film dispersion method, detergent method, injection method, microjet dispersion method, homogenizer facture, ultrasound wave dispersion method, freeze-drying, freeze-thaw method, single phase soln (such as tertiary butanol and water) method etc.The method that the outer aqueous phase substance of prepared blank liposome is removed or replaced comprises ultrafiltration, dialysis, sieve method, ion exchange resin (fiber) method etc.Comprehensive it, basic technology of the present invention is: 1) first with citrate buffer solution (pH2～4) the aquation liposome membrane material of 100mM～1000mM, reduce particle diameter, the preparation blank liposome; 2) remove or replace the material of outer water, set up gradient; 3) add alkaline matter, such as sodium carbonate, sodium bicarbonate, sodium phosphate, regulate pH to 6.5～9.5, and add berberine or berberine citrate, berberine sulfate, berberine hydrochloride, hatch 5～30min for 40～70 ℃.
Also optionally comprise other known in pharmaceutical field material in the liposomal pharmaceutical preparation of the present invention, comprise isoosmotic adjusting agent, antiseptic, antioxidant etc., material wherein such as trehalose, sucrose, histidine, lactose, mannitol, xylitol, glucose, sodium chloride, protein etc., can be a kind of, also can be more than one.
Liposomal pharmaceutical preparation can be measured the envelop rate of medicine in liposome by routine techniques.The liposome encapsulation method for measuring has ultrafiltration, dialysis, column chromatography, microtrabeculae centrifuging etc., and all there is some problems in these methods.We adopt ion exchange resin (fiber) to carry out entrapment efficiency determination, have weak point consuming time, simple to operate, cheap, reusable, do not need the expensive advantages such as instrument and equipment.
Advantage of the present invention
The liposome encapsulation that different membrane materials form is all greater than 80%, even reaches 100%, process stabilizing.
The specific embodiment
Below in conjunction with embodiment, be described more specifically content of the present invention.Should be appreciated that enforcement of the present invention is not limited to the following examples, any pro forma accommodation and/or change that the present invention is made all will fall into protection domain of the present invention.
Contrast test
Carry out the berberine liposome medicine carrying according to document (Preparation of berberine hydrochloride liposomes by active loading method Chinese Pharmaceutical Journal 01 phase in 2004), the as a result difficult repetition of envelop rate, possible cause is the differences such as phospholipid composition, liposome structure.Continue further investigation and find that envelop rate is difficult to the problem of repetition, can solve by removing outer water citrate.Prepare same liposome according to this article method, with ion exchange resin (D331 type Cl
-The type anion) remove citrate, sodium bicarbonate is regulated pH to 6.8, and envelop rate is greater than 80%, and can reappear, and pH increases, and envelop rate is greater than 90%.
Embodiment 1
Prescription HSPC 3g
CH 1g
PEG-DSPE 1g
Aquation medium: 300mM citric acid soln (pH 4.00) 100ml.
The preparation blank liposome: the HSPC(hydrogenated soy phosphatidyl choline that takes by weighing recipe quantity), the CH(cholesterol), the PEG-DSPE(PEG molecular weight is 2000), 55 ℃, with 10ml dissolve with ethanol film material, get the lipid phase; The aquation medium that is preheated to 55 ℃ is injected the lipid phase, hatch 10 min, make the liposome first product, process through the 20000psi high pressure homogenize again, reduce the liposome particle diameter to being about 100nm, by the microporous filter membrane of 0.8,0.45,0.22 μ m, namely get blank liposome successively.
Set up the gradient liposome: calculate the 1.2ml(apparent volume that wets according to exchange capacity) resin anion (R.A.) (OH
-) (717 type resin anion (R.A.)) namely can complete exchange 300mmol/L citric acid soln 1ml.Set up gradient by ion exchange resin, obtain the gradient blank liposome.
Medicine carrying initiatively: get gradient liposome 10mL, add sodium bicarbonate solution and regulate pH to about 8, add 10mL berberine hydrochloride solution (2mg/ml), 60 ℃ of insulation 10 min make berberine liposome.Particle diameter is 126nm, and envelop rate is about 96%.
The formulation and technology of present embodiment can obtain the different-grain diameter liposome by reducing dispersive pressure.
Also optionally comprise other known in pharmaceutical field material in this liposomal pharmaceutical preparation, comprise isoosmotic adjusting agent, antiseptic, antioxidant etc., material wherein such as trehalose, sucrose, histidine, lactose, mannitol, glucose, xylitol, sodium chloride, protein etc., can be a kind of, also can be more than one.
Still targeting material lipid derivate be can add in this liposome membrane material, such as targeting antibodies, part, transferrins, lactoferrin, folic acid, galactose, glucose, RGD etc. comprised.Still comprise cell membrane penetrating peptide lipid derivate etc.
Embodiment 2
Prescription
DSPC 0.9g
CH 0.3g
Tocopherol 0.01g
PEG5000-DSPE 0.3g
Get an amount of chloroform dissolving DSPC(distearoyl phosphatidylcholine), the CH(cholesterol), tocopherol, PEG5000-DSPE, rotary evaporation is removed chloroform, the preparation lipid film.Take 50ml 300mM citrate buffer (pH2.5) as the aquation medium, hydrated phospholipid is thin, the preparation large unilamellar vesicle.Process through the 30000psi high pressure homogenize, reduce the liposome particle diameter to being about 60nm, by the microporous filter membrane of 0.8,0.45,0.22 μ m, namely get blank liposome successively.
Set up the gradient liposome: adopt ZB-2 type Cl
-The type anion-exchange fibre is removed outer water citric acid cushion, obtains the gradient blank liposome.
Medicine carrying initiatively: get gradient liposome 10mL, add sodium carbonate liquor and regulate pH to about 8.5, add 5mL berberine citrate solution (10mg/ml), 60 ℃ of insulation 20 min make berberine liposome.Particle diameter is 63nm, and envelop rate is about 91%.
Still targeting material lipid derivate be can add in this liposome membrane material, such as targeting antibodies, part, transferrins, lactoferrin, folic acid, galactose, glucose, RGD etc. comprised.Still comprise cell membrane penetrating peptide lipid derivate etc.
Embodiment 3
Prescription
HEPC 1g
CH 0.4g
Tocopherol hemisuccinic acid ester 0.3g
PEG200-DSPE lipid derivate 0.3g
Operational approach is with " embodiment 1 ".The gradient method for building up is for adopting the outer water of 5% xylitol solution dialysis patient ultrafiltration exchange.
Also optionally comprise other known in pharmaceutical field material in the liposomal pharmaceutical preparation of the present invention, comprise isoosmotic adjusting agent, antiseptic, antioxidant etc., material wherein such as trehalose, sucrose, histidine, lactose, mannitol, glucose, sodium chloride, protein etc., can be a kind of, also can be more than one.
Certainly, still can add the materials such as sodium bicarbonate, sodium phosphate, sodium hydrogen phosphate, sodium hydroxide, amine butantriol, Portugal's first ammonia, basic amino acid and regulate outer water pH to 6.5～9.5.
Still targeting material lipid derivate be can add in this liposome membrane material, such as targeting antibodies, part, transferrins, lactoferrin, folic acid, galactose, glucose, RGD etc. comprised.Still comprise cell membrane penetrating peptide lipid derivate etc.
Embodiment 4
Prescription
DPPC 1g
CH 0.4g
PEG2000-DPPE lipid derivate 0.3g
Lipid phase: get each lipid of above-mentioned prescription, add 5ml ethanol, dissolve to get the lipid phase in 65 ℃.
Water: the citrate buffer solution 30mL of pH2.8 of preparation 200mM is preheated to 65 ℃ and get final product.
Blank liposome preparation: with water be added to lipid mutually in, dispersed with stirring, small-sized nanometer machine is processed (20000psi), by 0.8 μ m, 0.45 μ m, 0.22 μ m microporous filter membrane, gets blank liposome turbid liquor successively.
Gradient is set up: remove the outer water citrate buffer solution of liposome with ultrafiltration, obtain having the blank liposome suspension of transmembrane gradient.
Medicine carrying: get above-mentioned gradient liposome 1 mL, add berberine sulfate solution (3 mg/mL) 1 mL, sodium carbonate liquor is regulated pH to 6.5, and 60 ℃ of insulation 10 min make berberine liposome.Particle diameter is 126nm, envelop rate 82%.
Similarly, regulate pH to 7.0,7.5,8.0,8.5,9.0,9.5,10.0 with sodium carbonate liquor, measure envelop rate, be respectively 89%, 87%, 92%, 92%, 99%, 95%, 73%.Hence one can see that, and pH surpasses 9.5, and envelop rate significantly reduces, and possible cause is phospholipids degradation, causes the bimolecular film permeability to increase.
Still targeting material lipid derivate be can add in this liposome membrane material, such as targeting antibodies, part, transferrins, lactoferrin, folic acid, galactose, glucose, RGD etc. comprised.Still comprise cell membrane penetrating peptide lipid derivate etc.
Embodiment 5
The liposome prescription
DMPG 1g
PEG400-CH 0.3g
Operational approach is with " embodiment 1 ".Envelop rate is greater than 80%.
Still targeting material lipid derivate be can add in this liposome membrane material, such as targeting antibodies, part, transferrins, lactoferrin, folic acid, galactose, glucose, RGD etc. comprised.Still comprise cell membrane penetrating peptide lipid derivate etc.
Embodiment 6
The liposome prescription
DLPG 1g
CH 0.4
PEG2000-CH 0.1g
Operational approach is with " embodiment 1 ".Envelop rate is greater than 90%.
Still targeting material lipid derivate be can add in this liposome membrane material, such as targeting antibodies, part, transferrins, lactoferrin, folic acid, galactose, glucose, RGD etc. comprised.Still comprise cell membrane penetrating peptide lipid derivate etc.
The test of embodiment 7 antitumor
The mice of 36 inoculation S180 tumors is divided into 6 groups at random, be normal saline matched group (NS), berberine hydrochloride solution group, berberine liposome group (the different-grain diameter liposome of embodiment 1, envelop rate is greater than 95%), the conventional liposome group (is comprised of SPC and CH, particle diameter 186nm), every group of 6 animals.Inoculation was calculated according to the 0th day the same day, and the 4th day begins respectively at intravenous administration (20mg/kg) successive administration 7 days after the inoculation.Animal is normally raised after the administration, and weighing every day Mouse Weight is observed the growth conditions of mice simultaneously.After the inoculation the 11st day, put to death animal, completely peel off Subcutaneous tumor, calculate tumour inhibiting rate, the results are shown in Table 1.
Table 1 different-grain diameter liposome tumor suppression result
| The solution group | Conventional liposome (186nm) | PEGization liposome (326nm) | PEGization liposome (163nm) | PEGization liposome (107nm) | PEGization liposome (82nm) | PEGization liposome (39nm) | |
| Tumour inhibiting rate (%) | 18.8 | 33.2 | 37.2 | 55.7 | 61.3 | 58.1 | 65.2 |
As seen from table, the tumour inhibiting rate of conventional liposome (contrast test conventional liposome) is low, and PEGization liposome particle diameter is during greater than 200nm, such as 326nm, tumour inhibiting rate is also lower, when PEGization liposome particle diameter during less than 200nm, such as 163nm, 107 nm, 82 nm, 39 nm, tumour inhibiting rate significantly improves.
The test of embodiment 8 antitumor
The mices of 36 inoculation S180 tumors are divided into 6 groups at random, i.e. normal saline matched group (NS), berberine hydrochloride solution group, berberine liposome group, every group of 6 animals.Inoculation was calculated according to the 0th day the same day, and the 4th day begins respectively at intravenous administration (20mg/kg) successive administration 7 days after the inoculation.Animal is normally raised after the administration, and weighing every day Mouse Weight is observed the growth conditions of mice simultaneously.After the inoculation the 11st day, put to death animal, completely peel off Subcutaneous tumor, calculate tumour inhibiting rate, the results are shown in Table 2.
The different prescriptions of table 2 form liposome tumor suppression result
| The solution group | Embodiment 1 | Embodiment 3 | Embodiment 4 | Embodiment 5 | |
| Tumour inhibiting rate (%) | 26.3 | 62.3 | 57.3 | 69.1 | 56.6 |
Embodiment 9 liposome stabilities
4～8 ℃ of preservations of refrigerator check content, particle diameter, envelop rate, the results are shown in Table 3.
Table 3 stability result
Claims (12)
1. berberine liposome, it is characterized in that, comprise film material and berberine, described film material is phospholipid bilayer, by phospholipid, cholesterol, polyethyleneglycol lipid derivates forms, liposome particle size is 30～200nm, envelop rate is greater than 80%, described film material is one or more phospholipid, described phospholipid be natural or synthetic phospholipid or above-mentioned substance by the material of common method hydrogenation, be selected from phosphatidylcholine, phosphatidyl glycerol, phosphatidic acid, PHOSPHATIDYL ETHANOLAMINE, Phosphatidylserine, phosphatidylinositols, sphingomyelin, cuorin, HSPC, HEPC, DSPC, DPPC, DMPC, DSPG, DPPG, DMPG; Polyethyleneglycol lipid derivates is selected from polyethyleneglycol modified DSPE, polyethyleneglycol modified DSPG, polyethyleneglycol modified cholesterol, independent or applied in any combination; The mass ratio of phospholipid, cholesterol and polyethyleneglycol lipid derivates is 1:0～0.4:0.01～0.3(w/w/w).
2. berberine liposome according to claim 1 is characterized in that, described polyethyleneglycol lipid derivates is PEG2000-DSPE.
3. berberine liposome as claimed in claim 1 is characterized in that, the molecular weight of Polyethylene Glycol PEG is 100-100000 dalton in the described polyethyleneglycol lipid derivates.
4. berberine liposome as claimed in claim 1 is characterized in that, the molecular weight of Polyethylene Glycol PEG is 1000-5000 dalton in the described polyethyleneglycol lipid derivates.
5. the preparation method of a berberine liposome as claimed in claim 1, it is characterized in that, adopt the citric acid gradient, the technique that outer water adds alkaline matter adjusting pH is loaded into liposome with medicine, its preparation technology is: with organic solvent dissolving film material, remove/or do not remove organic solvent; Add citrate buffer solution hydration shell material, preparation liposome crude product; Reduce the liposome particle diameter, remove outer water citrate; Add alkaline matter and regulate pH, medicine carrying gets final product.
6. the preparation method of berberine liposome as claimed in claim 5 is characterized in that, the concentration of citrate buffer solution is 150mM～500mM; The alkaline matter that adds is selected from carbonate, phosphoric acid salt, sodium hydroxide, amine butantriol, Portugal's first ammonia, basic amino acid; Regulate outer water pH to 6.5～9.5.
7. the preparation method of berberine liposome as claimed in claim 6 is characterized in that, the alkaline matter that adds is selected from sodium bicarbonate, sodium phosphate or sodium hydrogen phosphate, regulates outer water pH7～9.
8. berberine liposome as claimed in claim 1 is characterized in that, also comprises antioxidant, isoosmotic adjusting agent, also comprises 10% with interior ethanol in preparation.
9. berberine liposome as claimed in claim 8 is characterized in that, described antioxidant is tocopherol, tocopherol hemisuccinic acid ester or EDTA; Described isoosmotic adjusting agent is sucrose, xylitol or trehalose.
10. berberine liposome as claimed in claim 1 is characterized in that, adds targeting material lipid derivate in membrane material.
11. berberine liposome as claimed in claim 10 is characterized in that, described targeting material lipid derivate is targeting antibodies or part.
12. berberine liposome as claimed in claim 11 is characterized in that, described targeting antibodies or part are selected from transferrins, lactoferrin, folic acid, galactose, glucose, RGD, albumin.
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| CN101108167A (en) * | 2007-08-03 | 2008-01-23 | 南京师范大学 | Method for preparing hydrochloric acid berberine long circulation liposome with ammonium sulphate gradient method-film evaporation method |
| CN101683322A (en) * | 2008-09-26 | 2010-03-31 | 华东理工大学 | Method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method |
| CN101804028A (en) * | 2010-04-20 | 2010-08-18 | 上海纳米技术及应用国家工程研究中心有限公司 | Method for preparing traditional Chinese medicine liposome |
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2010
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1446534A (en) * | 2002-12-30 | 2003-10-08 | 沈阳药科大学 | Lipoidosis of Chinese herbal medicine alkaloid and its preparation |
| CN101108167A (en) * | 2007-08-03 | 2008-01-23 | 南京师范大学 | Method for preparing hydrochloric acid berberine long circulation liposome with ammonium sulphate gradient method-film evaporation method |
| CN101683322A (en) * | 2008-09-26 | 2010-03-31 | 华东理工大学 | Method for preparing nano berberine hydrochloride liposome by supercritical carbon dioxide method |
| CN101804028A (en) * | 2010-04-20 | 2010-08-18 | 上海纳米技术及应用国家工程研究中心有限公司 | Method for preparing traditional Chinese medicine liposome |
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| 《主动载药法制备盐酸小檗碱脂质体》;邓意辉;《中国药学杂志》;20040131;第39卷(第1期);全文 * |
| 《盐酸小檗碱脂质体制备工艺研究》;苏春梅;《中国药业》;20091231;第18卷(第21期);全文 * |
| 苏春梅.《盐酸小檗碱脂质体制备工艺研究》.《中国药业》.2009,第18卷(第21期), |
| 邓意辉.《主动载药法制备盐酸小檗碱脂质体》.《中国药学杂志》.2004,第39卷(第1期), |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109602707A (en) * | 2019-02-03 | 2019-04-12 | 隋新兵 | Erussian liposome composition and preparation method thereof |
| CN109602707B (en) * | 2019-02-03 | 2021-09-03 | 隋新兵 | Erianin liposome composition and preparation method thereof |
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