CN105214091A - Multi-functional gold nano flower pesticide thing composite carrier and preparation method thereof - Google Patents
Multi-functional gold nano flower pesticide thing composite carrier and preparation method thereof Download PDFInfo
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Abstract
The present invention relates to a kind of multi-functional gold nano flower pesticide thing composite carrier and preparation method thereof, this medicine complex carrier is Jenner's popped rice with near infrared absorption function is core, in its Surface coating layer of silicon dioxide, then on silicon dioxide layer coating mesoporous silicon dioxide layer and the nanoparticle formed.Carrier material feature of the present invention is that silicon bi-layer shell is coated, outer mesoporous silicon oxide can be used for the medicines such as load Treated with Chemotherapeutic Drugs thing, optical dynamic therapy, there is universality, the direct coated Jenner's popped rice of internal layer silicon dioxide, avoid the ripening of Jenner's popped rice, ensure that strong mechanical property, Jenner's popped rice with near-infrared photo-thermal converting function can realize the drug controllable release under near infrared light, can be realized the synergism of chemotherapy, optical dynamic therapy and near infrared light thermal therapeutical by the medicine that load is different simultaneously.
Description
Technical field
The present invention relates to the targeted drug carrier material for disease treatment, be specifically related to a kind of irradiate by near-infrared laser under realize the golden nanometer particle composite carrier and preparation method thereof of the controllable release of medicine and heat, change therapeutic alliance effect.
Background technology
The targeted of medicine and controllable release are medical circle study hotspots always, and along with nano material technology development, nanoscale Metaporous silicon dioxide material becomes noticeable drug carrier material.Generally use the mode medicine carrying such as surface adsorption, surface functionalities group (as acrylic) covalently bound medicine at present, for reaching good medicine controlled releasing, realize drug release by temperature change, the change of environment pH value, the effect of pathological tissues peripheral protein enzyme etc.
The near-infrared thermotherapy of nanometer gold mediation, it is the photo-thermal Therapeutic Method in recent years occurred with the discovery of nanometer gold near infrared light hot-cast socket characteristic, nano Au particle is along with the change of size and surface topography, there is surface and wait particle resonance change, near infrared absorption function is provided with in near-infrared region, and can be rapidly that heat energy causes local temperature rising by light energy conversion, tumor cell necrosis apoptosis can be caused.Current researcher has prepared the particle such as nano ball shell, gold nanorods, gold nanometer cage, Jenner's popped rice that can be used for near-infrared thermotherapy.In order to reach better tumor thermotherapy effect, people attempt nanometer gold near infrared light thermal therapeutical to combine with chemotherapy, optical dynamic therapy etc., improve curative effect, and the research and development therefore as the multi-functional complex carrier of mediation material become crucial.The multifunctional nano gold combination drug carrier material of current bibliographical information has several as follows:
1. gold nano-spherical shell: with nano silicon spheres, hollow Nano liposome/silicon oxide ball etc. for core, the coated thin au shell in outside, by absorption chemotherapeutics or finishing acrylic, amino Deng Guanneng group connection medicine;
2. nanometer gold cage: hollow Nano gold cage, carboxylated etc. by finishing, connect medicine;
3. nanometer gold bar: connect medicine by the sense group of finishing, or by Shell Materials absorption medicines such as Surface coating porous silicon, Graphenes;
4. gold nano star or Jenner's popped rice: Jenner's popped rice is used for Raman signal to be strengthened, with during for pharmaceutical carrier, report is had directly to be mixed by the reactants such as photo-dynamical medicine methylene blue etc. and ethyl orthosilicate (TEOS), the nano-silicon Shell Materials of direct drug containing is prepared in co-precipitation, is coated on Jenner's popped rice surface.
Above-mentioned multi-functional medicament-carried nano complex carrier, for reaching its targeting, finishing monoclonal antibody while of normal, join fit, small peptide, saccharide, micromolecule etc., medicine realizes the selectivity release of medicine by the change of surrounding pH value, temperature change etc., but the nanometer gold combination drug carrier of preparation at present still also exists shortcomings such as lacking universality, drug release poor controllability, the improvement of nano Au particle stability needs.
Summary of the invention
What the object of the invention is to develop a kind of stable performance, can realize near-infrared controllable release and have a universality can the multifunctional nano composite carrier of load multi-medicament.
The present invention's multi-functional gold nano flower pesticide thing composite carrier is Jenner's popped rice with near infrared absorption function is core, in its Surface coating layer of silicon dioxide, then on silicon dioxide layer coating mesoporous silicon dioxide layer and the nanoparticle formed.
Described Jenner's popped rice particle diameter is 50 ~ 70nm, and silicon dioxide layer thickness is 20 ~ 30nm, and meso-porous titanium dioxide silicon layer thickness is 30 ~ 50nm.
The preparation method of the present invention's multi-functional gold nano flower pesticide thing composite carrier, comprises the following steps:
A. use Frens legal system for gold nano seed:
Under agitation the sodium citrate aqueous solution of 1% concentration is joined in the tetra chlorauric acid aqueous solution of the 0.25mM of heat boiling, the mol ratio of sodium citrate and tetra chlorauric acid is 7:1, until solution becomes burgundy color, after in 15min, color remains unchanged, stop heating and stir, centrifugalize after natural cooling, namely obtains precipitate color of spherical gold;
B. the preparation of Jenner's popped rice:
1) the gold nano seeded dispersion obtained by step a, in the sodium citrate aqueous solution of 1.75mM concentration, makes its concentration be 5.18 × 10
-11mol/L;
2) above-mentioned gold nano seed solution and aqueous ascorbic acid is got, in molar ratio for 200:1 prepares mixed liquor, and be 7 by this mixed liquor adjustment pH value, the amount being 200:20 by gold nano seed and tetra chlorauric acid mol ratio is afterwards on the waiting list tetra chlorauric acid aqueous solution, and to adjust pH value be 7;
3) under fast stirring, above-mentioned tetra chlorauric acid aqueous solution is joined fast in gold nano seed and ascorbic acid mixed liquor, now gold seeds in system: tetra chlorauric acid: ascorbic acid mol ratio is 200:20:1, solution becomes rapidly aeruginous Jenner popped rice solution, centrifugalize, namely obtains precipitate Jenner popped rice particle (summary is AuNF);
C. the ground floor of Jenner's popped rice is coated:
Dose volume ratio is TEOS:CH3CH2OH:H2O:NH3.H2O=1:2000:500:50
solution, the Jenner's popped rice particle getting step b acquisition joins in the stober solution of above-mentioned preparation, and Room-temperature seal stirs 3h ~ 4h, centrifugalize, namely obtains Jenner's popped rice particle (summary is AuNFSiO) with coated with silica layer;
D. the second layer of Jenner's popped rice is coated:
1) preparation of porous silicon coating reaction solution: after CH3CH2OH, H2O, NH3.H2O, TEOS 600:1400:5:3 mixing by volume, adds Organic substance and touches an edition CATB to make it concentration in system be 2 ~ 3mmol/L;
2) the Jenner's popped rice particle with coated with silica layer obtained by step c uses water and washing with alcohol twice respectively, join in above-mentioned porous silicon coating reaction solution, room temperature lower seal stirs centrifugalize after 3h ~ 4h, nanoparticle precipitate is used respectively water and washing with alcohol twice for subsequent use;
3) 6g/L ammonium nitrate/alcoholic solution is prepared, above-mentioned nanoparticle precipitate is distributed in ammonium nitrate/alcoholic solution, nanoparticle precipitate and ammonium nitrate mass ratio is made to be 1:120 ~ 1:60, reflux 6h to remove organic formwork CTAB under 60 DEG C of water-baths, be cooled to room temperature centrifugal, use water respectively, ethanol washes twice, and (summary is AuNFSiOmSiO namely to obtain the compound Jenner popped rice particle with mesoporous silicon oxide second clad
2).
This composite carrier not only can load multi-medicament, is irradiated and realizes drug controllable release, have near-infrared thermotherapeutic function simultaneously by near-infrared laser.The ideal carrier that the various medicine of load realizes thermochemotherapy can be widely used as.
The invention has the advantages that:
1. devise with Jenner's popped rice be core, the coated novel structure of double-deck silicon oxide shell, the coated Jenner's popped rice of internal layer oxide thin layer silicon shell, outer mesoporous silicon Shell Materials is coated can high-efficient carrier medicine;
2. ensure that the stability of gold nano floral material, for avoiding the ripening of Jenner's popped rice particle, the macromolecular compound that uses of preparation in the past as dispersant more, Jenner's popped rice that this research is prepared from aqueous phase, first coated oxide thin layer silicon materials, ensure that its stability and the function such as near infrared light hot-cast socket, Raman signal enhancing;
3. the stability of gold nano floral material ensure that the near-infrared photo-thermal controlled-release realizing medicine better;
4. carrier material of the present invention has universality, and outer mesoporous silicon shell material can load chemotherapeutics, photo-dynamical medicine etc., the different types of monoclonal antibody of modified surface, joins fit, small peptide etc. with the targeting demand of satisfied different cell tissue.
5. the inventive method with tetra chlorauric acid aqueous solution for growth solution, the Jenner's popped rice particle size prepared is even, good dispersion degree is not easily reunited, good stability, and do not use in preparation process there is Cytotoxic surfactant, improve the biological safety of material.Observe Jenner's popped rice particle under transmission electron microscope and record diameter about 50-70nm.Record the absorption spectrum of Jenner's popped rice through ultraviolet-visible spectrophotometer, main absworption peak, between 630-680nm, has band edge to lift in the near infrared region of more than 700nm, illustrates that this kind of gold nano floral material has certain near infrared absorption function.
6. by coated for Jenner's popped rice one deck thin silicon dioxide, silicon dioxide average thickness is about 20 ~ 30nm, Jenner's popped rice and surrounding not only can be protected to isolate, prevent ripening and reunion, and this core-shell structure of AuNFsSiO2 has better water solublity; The meso-porous titanium dioxide silicon thickness on top layer is approximately 30-50nm, have biocompatibility better, specific surface area is comparatively large, can carry out the features such as finishing
Accompanying drawing explanation
Fig. 1 is multi-functional gold nano flower pesticide thing complex carrier (AuNFSiO prepared by embodiment 1
2mSiO
2) transmission electricity photo;
Fig. 2 is AuNF, the AuNFSiO of preparation in embodiment 1
2, AuNFSiO
2mSiO
2particle uv-visible absorption spectra figure;
Fig. 3 is the AuNFSiO of variable concentrations
2mSiO
2photothermal conversion figure under different near-infrared intensity of illumination (power); Wherein: A, B, C power is respectively 5w/cm
2, 7w/cm
2and 9w/cm
2;
Fig. 4 be embodiment 1 multi-functional gold nano flower pesticide thing composite carrier load chemotherapeutic drugs Doxorubicin (DOX) of preparing afterwards under near infrared light, the asynchronous drug release rate schematic diagram of pH value.
Detailed description of the invention
By the following embodiment provided, the present invention is described in further details, but can not be interpreted as concrete restriction of the present invention.
Embodiment 1
The preparation of multi-functional gold nano flower pesticide thing complex carrier.
A. use Frens legal system for gold nano seed:
Under agitation the sodium citrate aqueous solution of 1% concentration is joined in the tetra chlorauric acid aqueous solution of the 0.25mM of heat boiling, the mol ratio of sodium citrate and tetra chlorauric acid is 7:1, until solution becomes burgundy color, after in 15min, color remains unchanged, stop heating and stir, centrifugalize after natural cooling, namely obtains precipitate color of spherical gold;
B. the preparation of Jenner's popped rice:
1) the gold nano seeded dispersion obtained by step a, in the sodium citrate aqueous solution of 1.75mM concentration, makes its concentration be 5.18 × 10
-11mol/L;
2) above-mentioned gold nano seed solution and aqueous ascorbic acid is got, in molar ratio for 200:1 prepares mixed liquor, and be 7 by this mixed liquor adjustment pH value, the amount being 200:20 by gold nano seed and tetra chlorauric acid mol ratio is afterwards on the waiting list tetra chlorauric acid aqueous solution, and to adjust pH value be 7;
3) under fast stirring, above-mentioned tetra chlorauric acid aqueous solution is joined fast in gold nano seed and ascorbic acid mixed liquor, now gold seeds in system: tetra chlorauric acid: ascorbic acid mol ratio is 200:20:1, solution becomes rapidly aeruginous Jenner popped rice solution, centrifugalize, namely obtains precipitate Jenner's popped rice particle (AuNF);
C. the ground floor of Jenner's popped rice is coated:
Dose volume ratio is TEOS:CH3CH2OH:H2O:NH3.H2O=1:2000:500:50
solution, the Jenner's popped rice particle getting step b acquisition joins in the stober solution of above-mentioned preparation, and Room-temperature seal stirs 3h ~ 4h, and centrifugalize, namely obtains Jenner's popped rice particle (AuNFSiO) with coated with silica layer;
D. the second layer of Jenner's popped rice is coated:
1) preparation of porous silicon coating reaction solution, after CH3CH2OH, H2O, NH3.H2O, TEOS 600:1400:5:3 mixing by volume, adds Organic substance and touches an edition CATB to make it concentration in system be 2 ~ 3mmol/L;
2) the Jenner's popped rice particle with coated with silica layer obtained by step c uses water and washing with alcohol twice respectively, join in above-mentioned porous silicon coating reaction solution, room temperature lower seal stirs centrifugalize after 3h ~ 4h, nanoparticle precipitate is used respectively water and washing with alcohol twice for subsequent use;
3) 6g/L ammonium nitrate/alcoholic solution is prepared, above-mentioned nanoparticle precipitate is distributed in ammonium nitrate/alcoholic solution, nanoparticle precipitate and ammonium nitrate mass ratio is made to be 1:120 ~ 1:60, reflux 6h to remove organic formwork CTAB under 60 DEG C of water-baths, be cooled to room temperature centrifugal, use water respectively, ethanol washes twice, namely obtains the compound Jenner popped rice particle (AuNFSiOmSiO with mesoporous silicon oxide second clad
2).
Prepared AuNFSiO
2mSiO
2particle, as shown in Figure 1, it is of a size of 150nm-200nm to the size modes under transmission electron microscope; AuNF, AuNFSiO
2, AuNFSiO
2mSiO
2record its absorption spectrum as shown in Figure 2 through ultraviolet-visible spectrophotometer, main absworption peak is positioned near 670nm.
Embodiment 2
To AuNFSiO prepared by embodiment 1
2mSiO
2the near infrared light hot-cast socket effect determination test of particle.
Use the Jenner popped rice compound particle of 808nm near-infrared laser to variable concentrations of different capacity to irradiate, power is respectively 5w/cm
2, 7w/cm
2and 9w/cm
2, sample concentration is respectively 0ug/ml, 100ug/ml, 200ug/ml, 300ug/ml and 400ug/ml, respectively gets 2ml, and each sample irradiates 20 minutes respectively.Fig. 3 is temperature change value in record experiment and draws near infrared light hot-cast socket real-time curve, and visible system temperature increases along with the increase of particle concentration and irradiation power.
Embodiment 3
To the biocompatibility test of AuNFSiO2mSiO2 particle composite carrier prepared by embodiment 1.
Use CCK-8 method to normal hepatocytes HL-7702 cell, people liver tumor HepG2 cell and human tongue carcinoma CAL27 cell measure.The method is easy and simple to handle, highly sensitive.Concentration is respectively 0 μ g/ml, 6.25 μ g/ml, 12.50 μ g/ml, 25 μ g/ml, the AuNFsSiO2mSiO2 of 50 μ g/ml and 100 μ g/ml joins in different cells and cultivates 24 little, and result carries out statistical analysis by SPSS16.0 software.Result of the test is as shown in table 1, table 2, table 3:
Table 1.CCK-8 testing inspection AuNFSiO
2mSiO
2research that nanoparticle affects Cal27 cell proliferation (n=6,
%)
Table 2.CCK-8 testing inspection AuNFSiO
2mSiO
2research that nanoparticle affects Hep-G2 cell proliferation (n=6,
)
Table 3.CCK-8 testing inspection AuNFSiO
2mSiO
2nanoparticle to the research of HL7702 cell proliferation (n=6,
%)
Along with the increase of effect metering, cell survival rate changes little each dosage group compared with Normal group, and difference does not have statistical significance (P ﹥ 0.05).Experiment proves, this new material does not have obvious cytotoxicity, has good biocompatibility.
Embodiment 4
The AuNFSiO2mSiO2 particle of embodiment 1 preparation is used as to the application test of pharmaceutical carrier.
(1) AuNFSiO
2mSiO
2the preparation of+DOX
Get amycin mother solution and draw amycin standard curve, concentration is 0.1mg/mL, is diluted to 10 μ g/mL respectively, 20 μ g/mL, 30 μ g/mL, 40 μ g/mL, 50 μ g/mL measure its ultraviolet-visible absorption spectroscopy, get the peak value at its absworption peak 480nm place, make UV, visible light standard absorption curve.
Get 2mg sample AuNFsSiO2mSiO2 and 3ml chemotherapeutic drugs Doxorubicin DOX (0.1mg/ml, PH=7.4), under room temperature, lucifuge stirs 12 hours.Then mixture is centrifugal and repeatedly wash twice by PBS solution, collect supernatant.Be determined wavelength by test DOX at 480nm ultraviolet characteristic absorption peak, according to the standard curve drawn at 480nm, test the absorbance of DOX, calculate quality and the concentration of DOX.In this experiment, the carrying drug ratio of carrier can reach 9.78%, and envelop rate is 62.6%.
(2) AuNFSiO
2mSiO
2the Drug Releasing Test of+DOX in different PH medium and under the irradiation of different near-infrared laser
By the support dispersion after medicine carrying in PBS (pH5.5 and the 7.4) solution of the different pH value of 2ml.Be placed in the water-bath of 37 DEG C respectively, get supernatant respectively at set intervals and by its quality of standard curve determination, and supplement the PBS solution of same volume.Calculate cumulative release amount, quality × 100% of medicine realeasing rate=Cumulative release amount/carrier Chinese medicine.808nm near-infrared laser (5W/cm is carried out to medicine carrying carrier
2) irradiate 20min, after having irradiated, collect supernatant rapidly, and measure its quality, calculate medicine realeasing rate, and curve plotting compares, as shown in Figure 4 under near-infrared irradiates, low PH can promote the release of medicine.
Claims (4)
1. a multifunctional drug composite carrier, is characterized in that, be Jenner's popped rice with near infrared absorption function is core, in its Surface coating layer of silicon dioxide, then on silicon dioxide layer coating mesoporous silicon dioxide layer and the nanoparticle formed.
2. multifunctional drug composite carrier according to claim 1, is characterized in that, described Jenner's popped rice particle diameter is 50 ~ 70nm, and silicon dioxide layer thickness is 20 ~ 30nm, and meso-porous titanium dioxide silicon layer thickness is 30 ~ 50nm.
3. prepare a method for multifunctional drug composite carrier described in claim 1, it is characterized in that comprising the following steps:
A. use Frens legal system for gold nano seed:
Under agitation the sodium citrate aqueous solution of 1% concentration is joined in the tetra chlorauric acid aqueous solution of the 0.25mM of heat boiling, the mol ratio of sodium citrate and tetra chlorauric acid is 7:1, until solution becomes burgundy color, after in 15min, color remains unchanged, stop heating and stir, centrifugalize after natural cooling, namely obtains precipitate color of spherical gold;
B. the preparation of Jenner's popped rice:
1) the gold nano seeded dispersion obtained by step a, in the sodium citrate aqueous solution of 1.75mM concentration, makes its concentration be 5.18 × 10
-11mol/L;
2) above-mentioned gold nano seed solution and aqueous ascorbic acid is got, in molar ratio for 200:1 prepares mixed liquor, and be 7 by this mixed liquor adjustment pH value, the amount being 200:20 by gold nano seed and tetra chlorauric acid mol ratio is afterwards on the waiting list tetra chlorauric acid aqueous solution, and to adjust pH value be 7;
3) under fast stirring, above-mentioned tetra chlorauric acid aqueous solution is joined fast in gold nano seed and ascorbic acid mixed liquor, now gold seeds in system: tetra chlorauric acid: ascorbic acid mol ratio is 200:20:1, solution becomes rapidly aeruginous Jenner popped rice solution, centrifugalize, namely obtains precipitate Jenner popped rice particle;
C. the ground floor of Jenner's popped rice is coated:
Dose volume ratio is TEOS:CH3CH2OH:H2O:NH3.H2O=1:2000:500:50
solution, the Jenner's popped rice particle getting step b acquisition joins in the stober solution of above-mentioned preparation, and Room-temperature seal stirs 3h ~ 4h, and centrifugalize, namely obtains Jenner's popped rice particle with coated with silica layer;
D. the second layer of Jenner's popped rice is coated:
1) preparation of porous silicon coating reaction solution: after CH3CH2OH, H2O, NH3.H2O, TEOS 600:1400:5:3 mixing by volume, adds Organic substance and touches an edition CATB to make it concentration in system be 2 ~ 3mmol/L;
2) the Jenner's popped rice particle with coated with silica layer obtained by step c uses water and washing with alcohol twice respectively, join in above-mentioned porous silicon coating reaction solution, room temperature lower seal stirs centrifugalize after 3h ~ 4h, nanoparticle precipitate is used respectively water and washing with alcohol twice for subsequent use;
3) 6g/L ammonium nitrate/alcoholic solution is prepared, above-mentioned nanoparticle precipitate is distributed in ammonium nitrate/alcoholic solution, nanoparticle precipitate and ammonium nitrate mass ratio is made to be 1:120 ~ 1:60, reflux 6h to remove organic formwork agent CTAB under 60 DEG C of water-baths, be cooled to room temperature centrifugal, use water respectively, ethanol washes twice, namely obtains the compound Jenner popped rice particle with mesoporous silicon oxide second clad.
4. multifunctional drug composite carrier according to claim 1, is preparing the application in near-infrared thermochemotherapy medicine.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108842270A (en) * | 2018-08-30 | 2018-11-20 | 佛山市南海区佳妍内衣有限公司 | It can three-dimensional structure fabric to fine tune temperature section |
| CN112775435A (en) * | 2020-12-30 | 2021-05-11 | 杭州电子科技大学 | Preparation method of gold nanorod surface-grown mesoporous silica material |
| CN112809016A (en) * | 2020-12-30 | 2021-05-18 | 杭州电子科技大学 | Preparation method of thickness-adjustable silicon dioxide material grown on surface of gold nanorod |
| CN112828284A (en) * | 2020-12-30 | 2021-05-25 | 杭州电子科技大学 | Preparation method of mesoporous silica-coated gold nanorod surface-grown silver composite material |
| CN113181139A (en) * | 2021-05-18 | 2021-07-30 | 首都医科大学附属北京友谊医院 | Method for preparing selenium/double-layer silicon dioxide spherical shell nano particles |
| CN113244198A (en) * | 2021-05-18 | 2021-08-13 | 首都医科大学附属北京友谊医院 | Selenium/double-layer silicon dioxide spherical shell nano particle and composition thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108842270A (en) * | 2018-08-30 | 2018-11-20 | 佛山市南海区佳妍内衣有限公司 | It can three-dimensional structure fabric to fine tune temperature section |
| CN112775435A (en) * | 2020-12-30 | 2021-05-11 | 杭州电子科技大学 | Preparation method of gold nanorod surface-grown mesoporous silica material |
| CN112809016A (en) * | 2020-12-30 | 2021-05-18 | 杭州电子科技大学 | Preparation method of thickness-adjustable silicon dioxide material grown on surface of gold nanorod |
| CN112828284A (en) * | 2020-12-30 | 2021-05-25 | 杭州电子科技大学 | Preparation method of mesoporous silica-coated gold nanorod surface-grown silver composite material |
| CN112809016B (en) * | 2020-12-30 | 2022-11-15 | 杭州电子科技大学 | Preparation method of thickness-adjustable silicon dioxide material grown on surface of gold nanorod |
| CN113181139A (en) * | 2021-05-18 | 2021-07-30 | 首都医科大学附属北京友谊医院 | Method for preparing selenium/double-layer silicon dioxide spherical shell nano particles |
| CN113244198A (en) * | 2021-05-18 | 2021-08-13 | 首都医科大学附属北京友谊医院 | Selenium/double-layer silicon dioxide spherical shell nano particle and composition thereof |
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Application publication date: 20160106 |