CN105078926A - Nano-carrier entrapping anticancer drugs and gold nanoparticles lipids and preparation method of nano-carrier - Google Patents
Nano-carrier entrapping anticancer drugs and gold nanoparticles lipids and preparation method of nano-carrier Download PDFInfo
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- CN105078926A CN105078926A CN201510529705.7A CN201510529705A CN105078926A CN 105078926 A CN105078926 A CN 105078926A CN 201510529705 A CN201510529705 A CN 201510529705A CN 105078926 A CN105078926 A CN 105078926A
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Abstract
The invention relates to a nano-carrier entrapping anticancer drugs and gold nanoparticles lipids and a preparation method of the nano-carrier, and belongs to the technical field of medicines. The nano-carrier entraps the anticancer drugs and the gold nanoparticles jointly and then is modified by functional molecules and (or) high molecular polymers. Draw ratio of gold nanoparticles in the carrier is 1:1-8:1, average grain diameters are 1 nanometer to 100 nanometers, and the temperature can rise to 25-80 DEG C after solution of the nano-carrier is irradiated by near-infrared light. The average grain diameter of the lipid nano-carrier is 20 nanometers to 500 nanometers. The lipid nano-carrier has effects of tumor targeted chemical treatment and/or tumor thermal synergy therapy, toxic and side effects of the anticancer drugs can be reduced, and an anti-tumor effect is improved. A preparation technology is simple, and the stability of the lipid nano-carrier is high.
Description
Technical field
The invention belongs to medical sci-tech field, be specifically related to common bag loaded with anti-cancer medicine and Jenner's grain of rice liposome nanometer carrier and preparation method thereof.
Background technology
Gold nanorods is due to the optics of its uniqueness and physicochemical character, and the application in tumor thermotherapy has become the focus that scientific research is paid close attention to.Along with the change of gold nanorods draw ratio, can regulate and control from visible region to near-infrared region, and near infrared light region wave-length coverage is the transmission window of body tissue, produces heat for utilizing gold nanorods by absorbing near infrared light and kill tumor cell and provide theoretical foundation.Gold nanorods, under the irradiation of laser, can absorb near-infrared low-energy radiation, makes its inner generation electron transition and forms unstable electron-hole pair, waiting electron-hole pair to reset when returning to steady statue and just can produce a large amount of heat.The heat produced can make cancerous cell temperature raise, and effectively can kill tumor cell, reaches the object for the treatment of tumor.
Liposome nanometer carrier is the more a kind of new drug carrier of Recent study, and it has phospholipid bilayer, with biomembrane bilayer similar, can wrap and carry fat-soluble, water soluble drug and thermotherapeutic agent etc.Fat-soluble medicine bag is downloaded to after in liposome, can increase the dissolubility of medicine, the action time of prolong drug, but also can change medicine distribution in vivo, substantially increases the drug level in tumor tissues.By the receptor-ligand of specific targets neoplastic cells or antibody modification liposome nanometer carrier, the targeted such as the medicine that bag can be loaded in wherein and thermotherapeutic agent play drug effect to tumor tissues.
Documents (CN103768600A) discloses a kind of magnetic thermal sensitive liposome nano-Au composite, and described complex is that thermal sensitive liposome is wrapped up Fe
3o
4after magnetic nano-particle and chemotherapeutics, then load gold nano grain is prepared from; This patent mainly applies Physical Target to (magnetic) delivering drugs and thermo-sensitive material, its poor biocompatibility, and targeting efficiency is low, and therefore therapeutic effect is not obvious low with safety.
Summary of the invention
The object of the invention is to the shortcoming overcoming tumor single therapy, a kind of construction method based on the common bag loaded with anti-cancer medicine of targets neoplastic cells and thermotherapeutic agent liposome nanometer carrier is provided, this liposome nanometer carrier can play the effect of cancer target chemotherapy and combined with hyperthermia or Synergistic treatment tumor, the toxic and side effects of cancer therapy drug can be reduced, improve Graft Versus Tumor.Preparation technology of the present invention is simple, liposome nanometer carrier good stability.
The present invention seeks to be achieved through the following technical solutions:
Bag loaded with anti-cancer medicine of the present invention and Jenner's grain of rice liposome nanometer carrier, in liposome nanometer carrier, bag is loaded with Jenner's grain of rice and cancer therapy drug, and described bag loaded with anti-cancer medicine and the finishing of Jenner's grain of rice liposome nanometer carrier have functional molecular and (or) high molecular polymer.
The draw ratio of described Jenner's grain of rice is 1:1 ~ 8:1, and mean diameter is 1nm ~ 100nm, and its solution temperature of near infrared light can be elevated to 25 DEG C ~ 80 DEG C.
Described cancer therapy drug is one or more in paclitaxel, Docetaxel, amycin, epirubicin, daunorubicin, idarubicin, pirarubicin, penta soft ratio, mitoxantrone, camptothecine, hydroxy camptothecin.
Described bag loaded with anti-cancer medicine and Jenner's grain of rice liposome nanometer carrier, wherein bag loaded with anti-cancer medicine and Jenner's grain of rice liposome nanometer carrier are liposome and solid lipid nanoparticle, and mean diameter is 20nm ~ 500nm.
Described functional molecular comprise can with one or more in the part of the specific antibody of tumor cell surface expression, ligand binding, antibody and aptamer, described high molecular polymer Polyethylene Glycol, mean molecule quantity is 1000 ~ 5000.
Described functional molecular is RLT polypeptide, folic acid, rgd peptide, TAT polypeptide, NGR polypeptide, SL
2one in B aptamer etc.
The preparation method of bag loaded with anti-cancer medicine of the present invention and Jenner's grain of rice liposome nanometer carrier is as follows: by ultrasonic dissolutions such as lipid material and medicines in organic solvent, decompression rotary evaporation flings to organic solvent, transparent membrane is formed in round-bottomed flask, add gold nanorods aqueous solution, continue to rotate aquation, ultrasonic, afterwards that it is centrifugal, discard precipitation, add functional molecular wherein again, room temperature shaker oscillating reactions, obtain loaded with anti-cancer medicine and Jenner's grain of rice liposome nanometer carrier.
Bag loaded with anti-cancer medicine of the present invention and Jenner's grain of rice liposome nanometer carrier have the effect of cancer target chemotherapy and combined with hyperthermia or Synergistic treatment tumor.
Adopt the common bag loaded with anti-cancer medicine of liposome nanometer carrier and Jenner's grain of rice, use again afterwards the functional molecular of targets neoplastic cells as with the specific antibody of tumor cell surface expression, the part of ligand binding, the modification such as antibody and aptamer, by cancer therapy drug and Jenner's grain of rice targeted delivery to tumor cell, the effect of cancer target chemotherapy and combined with hyperthermia or Synergistic treatment tumor can be played.
Liposome nanometer carrier finishing of the present invention has efficient targeting and has functional molecular and (or) the high molecular polymer of good biocompatibility, can significantly strengthen cancer target efficiency, improve the efficiency of tumor thermotherapy chemotherapy combined treatment, and reducing the toxic and side effects of its normal tissue organ, safety is high.
Advantage of the present invention is: present invention utilizes the Chemotherapy of cancer therapy drug and the thermotherapy characteristic of Jenner's grain of rice, achieve the effect of cancer target chemotherapy and combined with hyperthermia or Synergistic treatment tumor.Cancer therapy drug and Jenner's grain of rice are wrapped jointly and is loaded in liposome nanometer carrier, improve the stability of medicine and Jenner's grain of rice, after target function molecular modification, in turn enhance the uptake ratio of its tumor-targeting and tumor cell.Therefore, overcome traditional single chemotherapy poor specificity, toxic and side effects is large, the shortcoming that selectivity is low.Preparation technology of the present invention is simple, liposome nanometer carrier good stability, is expected to become the clinical treatment that a kind of new Therapeutic Method is applied to cancer.
Accompanying drawing explanation
Fig. 1 is the transmission electron microscope picture of gold nanorods;
Fig. 2 is gold nanorods spectrogram and temperature changing trend corresponding under laser irradiates thereof;
Fig. 3 is the transmission electron microscope picture of AuNRs/DTXLNC-RLT;
Fig. 4 is the release in vitro feature of AuNRs/DTXLNC-RLT;
Fig. 5 is the intensification experiment of AuNRs/DTXLNC-RLT;
Fig. 6 is the inhibitory action of AuNRs/DTXLNC-RLT to PC-3 tumor cell proliferation.
Detailed description of the invention
By cholesterol, soybean phospholipid, hydrogenated phospholipid, cancer therapy drug, DSPE-PEG (2000)-Mal ultrasonic dissolution is in chloroform, decompression rotary evaporation flings to chloroform, transparent membrane is formed in round-bottomed flask, add 4ml gold nanorods aqueous solution, continue to rotate aquation, aquation completely after ultrasonic dissolution again, in ultrasonic cell disruptor, ice bath is visited and is surpassed 40 times, by its centrifugal 10min under 8000r/min condition, discard precipitation, add functional molecular aqueous solution wherein again, vortex 1min, room temperature shaker oscillating reactions 12h, obtain bag loaded with anti-cancer medicine and Jenner's grain of rice liposome nanometer carrier, instantiation is as following table, but be not limited thereto.
| Bag loaded with anti-cancer medicine and Jenner's grain of rice liposome nanometer carrier | Cancer therapy drug | Functional molecular |
| AuNRs/TaxLNC-RLT | Paclitaxel (Tax) | RLT |
| AuNRs/DOXLNC-RGD | Amycin (DOX) | RGD |
| AuNRs/DTXLNC-TAT | Docetaxel (DTX) | TAT |
| AuNRs/CPTLNC-NGR | Camptothecine (CPT) | NGR |
| AuNRs/HCPTLNC-SL 2B | Hydroxy camptothecin (HCPT) | SL 2B |
| AuNRs/DTXLNC-RLT | Docetaxel (DTX) | RLT |
| AuNRs/TaxLNC-RGD | Paclitaxel (Tax) | RGD |
Now for Docetaxel (DTX) conventional clinically and low density lipoprotein, LDL peptide (RLT), set forth detailed description of the invention.
the preparation of Jenner's grain of rice
The preparation of Jenner's grain of rice seed: be that the cetyl trimethyl ammonium bromide aqueous solution of 0.2mol/L is placed in 25 ~ 26 DEG C of thermostat water baths by concentration, and to add concentration be wherein 5 × 10
-4the aqueous solution of chloraurate of mol/L, be uniformly mixed rear solution and become brown color, in above-mentioned solution, add the sodium borohydride aqueous solution that concentration is the ice of 0.01mol/L again, be statically placed in after rapid stirring 2min in the water-bath at this temperature, obtain dark brown Jenner's grain of rice seed solution.Wherein, the mol ratio of the aqueous solution mixture of the aqueous solution of cetyl trimethyl ammonium bromide, the aqueous solution of gold chloride and sodium borohydride is 1:0.0025:0.006.
The preparation of growth-promoting media: insert the cetyl trimethyl ammonium bromide aqueous solution that concentration is 0.2mol/L in 25 ~ 26 DEG C of thermostat water baths, add concentration 4 × 10
-3the silver nitrate aqueous solution of mol/L, adding concentration after stirring again is wherein 1 × 10
-3the aqueous solution of chloraurate of mol/L, the solution that stirs becomes brown color from colourless, then adds the aqueous ascorbic acid that concentration is 0.0788mol/L in this mixed solution, and rapid stirring becomes colorless to this mixed solution, is called growth solution.Wherein, the mol ratio of cetyl trimethyl ammonium bromide aqueous solution, silver nitrate aqueous solution, aqueous solution of chloraurate, aqueous ascorbic acid mixture is 1:0.0008:0.005:0.005516.
The preparation of gold nanorods: add a certain amount of seed solution in growth-promoting media, be positioned in 27 ~ 30 DEG C of water-baths after mix homogeneously, within about 5 minutes, this solution becomes kermesinus from colourless, continue to be positioned over 12h in the water-bath under this condition, obtain gold nanorods aqueous solution, wherein, seed solution is 11:102.7 with the ratio of the volume of growth-promoting media, and form and the size of gold nanorods are shown in Fig. 1.
the temperature rise effect of Jenner's grain of rice
The gold nanorods of same concentrations, its longitudinal absworption peak peak position is respectively the gold nanorods of 720nm, 749nm, 804nm, 843nm under laser irradiates, its temperature raises along with the increase of laser irradiation time, and during irradiation 8min, its temperature reaches 46 DEG C, 50 DEG C, 54 DEG C, 59 DEG C respectively, sees Fig. 2.Gold nanorods has surface plasma body resonant vibration wavelength, the near-infrared low-energy radiation that continuous-wave laser is launched can be absorbed, gold nanorods inside is made to produce electron transition and form unstable electron-hole pair, when electron-hole pair returns to steady statue, two-photon fluorescence and a large amount of heat will be produced.The gold nanorods of identical longitudinal absworption peak peak position is under laser irradiates, and it is higher that the larger temperature of concentration rises.Result shows, longitudinal absworption peak peak position is that the gold nanorods Thermogenesis of 804nm is the strongest, and has time and concentration dependent.
the common bag loaded with anti-cancer medicine of cancer target modified and the preparation of thermotherapeutic agent liposome nanometer carrier (AuNRs/DTXLNC-RLT)
By cholesterol, soybean phospholipid, hydrogenated phospholipid, DTX, DSPE-PEG (2000)-Mal ultrasonic dissolution in chloroform, decompression rotary evaporation flings to chloroform, transparent membrane is formed in round-bottomed flask, add 4ml gold nanorods aqueous solution, continue to rotate aquation, aquation completely after ultrasonic dissolution again, in ultrasonic cell disruptor, ice bath is visited and is surpassed 40 times, by its centrifugal 10min under 8000r/min condition, discard precipitation, then add RLT polypeptid solution wherein, vortex 1min, room temperature shaker oscillating reactions 12h, obtains AuNRs/DTXLNC-RLT.
Adopt that laser nano Particle Size Analyzer measures the particle diameter of AuNRs/DTXLNC-RLT, PDI and zeta current potential is respectively 163.15 ± 1.83nm, 0.25 ± 0.04 and ﹣ 32.8 ± 2.16mV.Result shows the even particle size distribution of AuNRs/DTXLNC-RLT, and stability is better.HPLC measures its content of dispersion and envelop rate is respectively 1.41 ± 0.11mg/ml and 98.45 ± 0.37%.Its configuration is shown in Fig. 3.
tablets in vitro experiment
The tablets in vitro of AuNRs/DTXLNC-RLT the results are shown in Figure 4.Connect RLT group obviously not distinguish with the rate of releasing drug not connecting RLT group.In identical release medium, the release dose rates of same preparation laser irradiation group is obviously accelerated.The main cause that laser irradiates the quickening of lower release dose rates may be that gold nanorods produces a large amount of heat under laser irradiates, and solution temperature raises the phase transition temperature reaching lecithin, and along with lecithin permeability changes, medicine is easier to be discharged from system.Be likely also that the large calorimetric that gold nanorods produces makes the temperature of whole medicine-carried system raise, drug molecule motion is more violent, accelerates its diffusion velocity in the solution, finally shows as the quickening of release dose rates.
intensification experiment
AuNRs/DTXLNC-RLT irradiates under 808 laser, measures its temperature, and the time dependent data of record temperature, and curve plotting figure, the results are shown in Figure 5.Along with the increase of laser irradiation time, the temperature of system constantly rises.Under identical laser irradiation condition, concentration is larger, and system temperature raises faster.Containing the carrier system of 0.2nM gold nanorods, laser irradiates 5min system temperature just can be elevated to 52 DEG C.This result illustrates, the photothermal deformation of carrier system is very capable, can be used for tumor thermotherapy.
antitumor action
AuNRs/DTXLNC-RLT the results are shown in Figure 6 to PC-3 Cytostatic to tumor cell rate.The suppression ratio of various preparation to PC-3 cell all shows concentration dependent, and the increase suppression ratio namely with formulation concentrations is increase trend.DTX liposome (DTXLNC) group is to the suppression ratio of cell apparently higher than crude drug DTX group, and AuNRs/DTXLNC group and DTXLNC group are suitable to the suppression ratio of cell, but AuNRs/DTXLNC group obviously increases the suppression ratio of cell under laser irradiates.AuNRs/DTXLNC-RLT group under laser irradiates with not with compared with laser irradiation group, it is more remarkable to the inhibitory action of tumor cell proliferation.
Claims (7)
1. a bag loaded with anti-cancer medicine and Jenner's grain of rice liposome nanometer carrier, it is characterized in that, in liposome nanometer carrier, bag is loaded with Jenner's grain of rice and cancer therapy drug, and described bag loaded with anti-cancer medicine and the finishing of Jenner's grain of rice liposome nanometer carrier have functional molecular and (or) high molecular polymer.
2. bag loaded with anti-cancer medicine according to claim 1 and Jenner's grain of rice liposome nanometer carrier, is characterized in that: the draw ratio of described Jenner's grain of rice is 1:1 ~ 8:1, and mean diameter is 1nm ~ 100nm, and its solution temperature of near infrared light can be elevated to 25 DEG C ~ 80 DEG C.
3. bag loaded with anti-cancer medicine according to claim 1 and Jenner's grain of rice liposome nanometer carrier, is characterized in that: described cancer therapy drug is one or more in paclitaxel, Docetaxel, amycin, epirubicin, daunorubicin, idarubicin, pirarubicin, penta soft ratio, mitoxantrone, camptothecine, hydroxy camptothecin.
4. bag loaded with anti-cancer medicine according to claim 1 and Jenner's grain of rice liposome nanometer carrier, is characterized in that: wherein bag loaded with anti-cancer medicine and Jenner's grain of rice liposome nanometer carrier are liposome and solid lipid nanoparticle, and mean diameter is 20nm ~ 500nm.
5. bag loaded with anti-cancer medicine according to claim 1 and Jenner's grain of rice liposome nanometer carrier, it is characterized in that: described functional molecular comprise can with one or more in the part of the specific antibody of tumor cell surface expression, ligand binding, antibody and aptamer, described high molecular polymer Polyethylene Glycol, mean molecule quantity is 1000 ~ 5000.
6. bag loaded with anti-cancer medicine according to claim 5 and Jenner's grain of rice liposome nanometer carrier, is characterized in that: described functional molecular is RLT polypeptide, folic acid, rgd peptide, TAT polypeptide, NGR polypeptide, SL
2one in B aptamer etc.
7. the preparation method of bag loaded with anti-cancer medicine according to claim 1 and Jenner's grain of rice liposome nanometer carrier: by ultrasonic dissolutions such as lipid material and medicines in organic solvent, decompression rotary evaporation flings to organic solvent, transparent membrane is formed in round-bottomed flask, add gold nanorods aqueous solution, continue to rotate aquation, ultrasonic, afterwards that it is centrifugal, discard precipitation, add functional molecular wherein again, room temperature shaker oscillating reactions, obtain loaded with anti-cancer medicine and Jenner's grain of rice liposome nanometer carrier.
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| CN108498460A (en) * | 2017-02-24 | 2018-09-07 | 国家纳米科学中心 | Gold nanoclusters-liposome composite particles and its preparation method and application |
| CN109044993A (en) * | 2018-09-18 | 2018-12-21 | 华南理工大学 | It is a kind of to target polyethyleneglycol modified mesoporous silicon dioxide nano particle and preparation method thereof with aptamer |
| CN114867472A (en) * | 2019-12-23 | 2022-08-05 | 巴斯克大学 | Gold lipid nanoparticles for gene therapy |
| CN114867472B (en) * | 2019-12-23 | 2024-04-19 | 巴斯克大学 | Gold lipid nanoparticles for gene therapy |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109044993A (en) * | 2018-09-18 | 2018-12-21 | 华南理工大学 | It is a kind of to target polyethyleneglycol modified mesoporous silicon dioxide nano particle and preparation method thereof with aptamer |
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Application publication date: 20151125 |