CN103893764A

CN103893764A - Drug carrier, preparation method thereof, pharmaceutical composition made from drug carrier, and applications of drug carrier and pharmaceutical composition

Info

Publication number: CN103893764A
Application number: CN201210572112.5A
Authority: CN
Inventors: 陈春英; 仉振江; 王静; 王黎明; 吴晓春
Original assignee: National Center for Nanosccience and Technology China
Current assignee: National Center for Nanosccience and Technology China
Priority date: 2012-12-25
Filing date: 2012-12-25
Publication date: 2014-07-02

Abstract

The invention discloses a drug carrier, a preparation method, a pharmaceutical composition made from the drug carrier, and applications of the drug carrier and the pharmaceutical composition. The mesoporous silica-coated gold nanorod (Au@SiO2) carrier is characterized in that: gold nanorods are coated with a layer of mesoporous silica. According to the preparation method, the Au@SiO2 carrier is prepared by taking hexadecyl trimethyl ammonium bromide-coated gold nanorods as raw materials via hydrolytic polymerization with orthosilicate ester, can be used for coating a plurality of drugs and probe molecules, and delivering the drugs and the probe molecules to parts of diseases so as to realize targeting therapy. After absorption of near-infrared laser, the Au@SiO2 carrier, or drug-loaded Au@SiO2 nanoparticles are capable of emitting fluorescence and transforming a part of light energy into heat energy, so that the Au@SiO2 carrier and the drug-loaded Au@SiO2 nanoparticles can be used for bioimaging, light-dependent control drug release (chemotherapy), and thermotherapy. The preparation method of the Au@SiO2 carrier is simple; the Au@SiO2 carrier can be used for a plurality of diagnosis and treatment methods, and the plurality of diagnosis and treatment methods can be combined, so that it is beneficial for avoiding defects of a single diagnosis and treatment method, and the drug carrier and the pharmaceutical composition are especially suitable for tumor diagnosis and treatment with complex requirements.

Description

Pharmaceutical carrier and preparation method thereof and pharmaceutical composition and application thereof

Technical field

The present invention relates to pharmaceutical carrier and preparation method thereof and pharmaceutical composition and application thereof.Particularly, relate to a kind of pharmaceutical carrier and preparation method thereof, a kind of on described pharmaceutical carrier load the pharmaceutical composition of medicine and described pharmaceutical carrier and the application of described pharmaceutical composition in the medicine for the preparation of diagnosis and/or treatment tumor.

Background technology

Traditional cancer treatment method, due to the targeting lacking for cancerous tissue, also can hurt non-pathological tissues when performance therapeutic effect, often has very large side effect.In the last few years, nanotechnology showed unique advantage strengthening the pharmacologically active of cancer therapy drug and reduce aspect toxicity.In cancerous issue, owing to having, rich blood vessel, blood vessel wall gap are wider, poor structural integrity and lymphatic return disappearance, and make macromole and lipid granule have high-permeability and anelasticity effect.Nano medication also can utilize this effect, is optionally enriched in (being called " passive targeting ") around cancerous tissue by blood circulation, can effectively improve like this cancer therapy drug active and reduce side effect.In addition, can also identify specifically cancerous cell or cancerous tissue with the Nano medication of targeted molecular, strengthen anticancer effect by active targeting.Nanotechnology has been classified as one of strategic objective of coming 10 years nanosecond medical science development for the clinical treatment of tumor by NIH in 2005.

In the exploration that recent years, Application of micron was studied in cancer diagnosis and treatment, gold nano grain shows the character and the advantage that attract people's attention.Due to its adjustable surface plasma resonance effect, golden nanometer particle not only has broad application prospects aspect cell imaging, can also become by photo-thermal transition effects the thermal source of localization when the laser irradiation.The heat producing on the one hand can be used for doing cancer thermotherapy, in the time that nanoparticle is used as anti-cancer medicament carrier, also can trigger the release of medicine on the other hand, realizes chemotherapy.Therefore, golden nanometer particle is likely embodied as picture, chemotherapy and three kinds of cancer diagnosis and treatment functions of thermotherapy in a carrier simultaneously.In addition, wherein each function also may synergism, produces new treatment pattern, such as medicine transmission and the thermotherapy of imaging mediation, or the combined therapy that concurs of chemotherapy and thermotherapy.Building the cancer that this multifunctional nano carrier is conducive to needing complex process treats targetedly.

For color of spherical gold, longitudinal surface plasma resonance effect of gold nanorods can be controlled by changing its draw ratio easily, makes its maximum absorption wavelength be positioned near infrared region.Because near infrared light can penetrate darker tissue, so the gold nanorods of realizing various functions by outside laser irradiation is particularly useful for diagnosis and the treatment of the shallow cancers such as breast carcinoma.

Although gold nanorods has good application prospect in the diagnosis of cancer and treatment, but still faces a problem: gold nanorods is difficult for carrying medicaments.

Summary of the invention

The object of the invention is to overcome the problem that gold nanorods in prior art is difficult for carrying medicaments, a kind of novel pharmaceutical carrier and preparation method thereof is provided, this novel pharmaceutical carrier contains gold nanorods, and drug loading amount is large.

The present invention also aims to provide a kind of pharmaceutical composition that contains above-mentioned novel pharmaceutical carrier.

The present invention also aims to provide the application of aforementioned pharmaceutical compositions in diagnosis and/or treatment tumor.

That is, the invention provides a kind of pharmaceutical carrier, wherein, this pharmaceutical carrier comprises gold nanorods, and is wrapped in the meso-porous titanium dioxide silicon layer on gold nanorods surface.

The present invention also provides a kind of preparation method of pharmaceutical carrier, wherein, the method comprises the following steps: in the gold nanorods aqueous solution that contains cetyl trimethyl ammonium bromide, under the condition that is 8.5-11 at pH value, add the alcoholic solution of esters of silicon acis to react, and carry out solid-liquid separation, obtain surface and be coated with the gold nanorods of meso-porous titanium dioxide silicon layer.

The present invention also provides a kind of pharmaceutical composition, and described pharmaceutical composition comprises above-mentioned carrier and loads on the medicine on above-mentioned carrier.

The present invention also provides said medicine carrier and the application of aforementioned pharmaceutical compositions in the medicine for the preparation of diagnosis and/or treatment tumor.

Pharmaceutical composition provided by the invention has diagnosis, the multiple functions such as chemotherapy and thermotherapy, and these functions can use in conjunction.

According to novel pharmaceutical carrier provided by the present invention, owing to being coated with meso-porous titanium dioxide silicon layer at gold nanorods, can effectively prevent that pharmaceutical carrier from reuniting in aqueous solution, and drug loading amount is large.In addition, the pharmaceutical composition obtaining after carrying medicament on this carrier can be realized the diagnosis of tumor simultaneously, and chemotherapy and two kinds of therapeutic modalities of thermotherapy.Wherein, the selection of two kinds of therapeutic modalities and concrete Therapeutic Method, can carry out targetedly according to the result of diagnosis; The result for the treatment of can be reported by the diagnostic function of carrier, to determine follow-up therapeutic scheme.Chemotherapy and two kinds of therapeutic modalities of thermotherapy combine utilization, contribute to overcome two kinds of therapeutic modalities shortcoming separately, obtain optimum therapeutic effect.

Other features and advantages of the present invention are described in detail the specific embodiment part subsequently.

Accompanying drawing explanation

The transmission electron microscope figure of the gold nanorods that Fig. 1 obtains for embodiment 1;

The transmission electron microscope figure of the pharmaceutical carrier that Fig. 2 obtains for embodiment 1;

The transmission electron microscope figure of the pharmaceutical carrier that Fig. 3 obtains for embodiment 3;

Fig. 4 is the related temperature-irradiation time curve chart of embodiment 5;

Fig. 5 is pharmaceutical carrier (the Au@SiO that the related bag of embodiment 6 has carried amycin ₂-DOX) release curve chart;

Fig. 6 is the release curve chart that the related bag of embodiment 7 has carried the pharmaceutical carrier of paclitaxel;

Fig. 7 is the related constitutional diagram of embodiment 8, and wherein, the A in Fig. 7 is amycin (DOX) and lysosome network for location altogether; B in Fig. 7 is carrier A u@SiO ₂with lysosome (Lyso) network for location altogether; C in Fig. 7 is carrier A u@SiO ₂with amycin (DOX) network for location altogether; D in Fig. 7 is carrier A u@SiO ₂, amycin (DOX) and lysosome (Lyso) network for location altogether;

Fig. 8 is the related constitutional diagram of embodiment 9, and wherein, the A in Fig. 8 is amycin and mitochondrion (Mito) network for location altogether; B in Fig. 8 is carrier A u@SiO ₂with mitochondrion (Mito) network for location altogether; C in Fig. 8 is carrier A u@SiO ₂with amycin (DOX) network for location altogether; D in Fig. 8 is carrier A u@SiO ₂, amycin (DOX) and mitochondrion (Mito) network for location altogether;

Fig. 9 is the related constitutional diagram of embodiment 10, and wherein, the A in Fig. 9 is amycin and lysosome (Lyso) network for location altogether; B in Fig. 9 is carrier A u@SiO ₂with lysosome (Lyso) network for location altogether; C in Fig. 9 is carrier A u@SiO ₂with amycin (DOX) network for location altogether; D in Fig. 9 is carrier A u@SiO ₂, amycin (DOX) and lysosome (Lyso) network for location altogether;

Figure 10 is the related constitutional diagram of embodiment 11, and wherein, the A in Figure 10 is amycin (DOX) and mitochondrion (Mito) network for location altogether; B in Figure 10 is carrier A u@SiO ₂with mitochondrion (Mito) network for location altogether; C in Figure 10 is carrier A u@SiO ₂with amycin (DOX) network for location altogether; D in Figure 10 is carrier A u@SiO ₂, amycin (DOX) and mitochondrion (Mito) network for location altogether;

Figure 11 is the trendgram of the related cellular uptake amycin (DOX) of embodiment 12;

Figure 12 is the graph of a relation of the related amycin of embodiment 13 (DOX) concentration and cell viability;

Figure 13 is mesoporous silicon oxide gold-covered nano rod carrier (the Au@SiO that the related bag of embodiment 13 has carried amycin ₂-DOX, concentration is in the amycin of load) graph of a relation of concentration and cell viability;

Figure 14 is the design sketch of the related laser controlling pharmaceutical composition release in cell of embodiment 14;

Figure 15 is the figure of the impact of thermotherapy on lysosome membrane integrity that represent the related laser irradiation of embodiment 15 induction;

Figure 16 represents that the related low power laser of embodiment 16 irradiates the figure of the impact of the chemotherapy that causes drug release on 7 kinds of tumor cell activity;

Figure 17 is the figure that represents the thermotherapy of the related high power laser light radiation-induced of embodiment 17 and the impact of the chemotherapy synergism of drug release on cell viability;

Figure 18 represents the figure of the related pharmaceutical composition of embodiment 18 in the in-house distribution of animal tumor.

The specific embodiment

Below the specific embodiment of the present invention is elaborated.Should be understood that, the specific embodiment described herein only, for description and interpretation the present invention, is not limited to the present invention.

Pharmaceutical carrier of the present invention comprises gold nanorods, and is wrapped in the meso-porous titanium dioxide silicon layer on gold nanorods surface.

According to pharmaceutical carrier of the present invention, under normal circumstances, the length of described gold nanorods can be 5-200 nanometer, and under preferable case, described gold nanorods length is 10-150 nanometer; Consider from being conducive to cellular uptake, more preferably described gold nanorods length is 15-120 nanometer; Further preferred described gold nanorods length is 20-100 nanometer; Further preferred described gold nanorods length is 30-80 nanometer.

More preferably, the draw ratio of described gold nanorods is 1.5-20; From being conducive to cellular uptake and being suitable for considering by Submillineter Wave Technology control, further the draw ratio of preferred described gold nanorods is 2-10; Further the draw ratio of preferred described gold nanorods is 2.5-6, most preferably 3-4.5.

According to pharmaceutical carrier of the present invention, described gold nanorods be coated with meso-porous titanium dioxide silicon layer.Under preferable case, the thickness of described meso-porous titanium dioxide silicon layer is 3-50 nanometer; Consider from being easy to preparation and being suitable for cellular uptake, more preferably the thickness of described meso-porous titanium dioxide silicon layer is 5-45 nanometer; Further the thickness of preferred described meso-porous titanium dioxide silicon layer is 10-40 nanometer; Further be preferably 15-35 nanometer.

Under preferable case, the aperture of described meso-porous titanium dioxide silicon layer is 1-20 nanometer; Consider from being easy to preparation and increasing stability, more preferably the aperture of described meso-porous titanium dioxide silicon layer is 1.2-15 nanometer; Further the aperture of preferred described meso-porous titanium dioxide silicon layer is 1.5-10 nanometer; It is further 2-8 nanometer; Further be preferably 3-4 nanometer.

More preferably, the specific surface area of described mesoporous pharmaceutical carrier is 1-150 meters squared per gram, pore volume be 0.05-10 cubic centimetre/gram; Increase stability and consider from being easy to preparation, further the specific surface area of preferred described pharmaceutical carrier is 30-120 meters squared per gram, pore volume be 0.5-5 cubic centimetre/gram; Further the specific surface area of preferred described pharmaceutical carrier is 60-100 meters squared per gram, pore volume be 0.6-2 cubic centimetre/gram; Further the specific surface area of preferred described pharmaceutical carrier is 65-95 meters squared per gram, pore volume be 0.65-1.3 cubic centimetre/gram.

According to pharmaceutical carrier of the present invention, under preferable case, its maximum absorption band is in the scope of 600-1500 nanometer, more preferably in the scope of 650-1000 nanometer.

There is the pharmaceutical carrier of the present invention of said structure, in aqueous solution, be not easy to reunite, and the drug loading amount of pharmaceutical carrier is large.

The present invention also provides a kind of preparation method of pharmaceutical carrier, wherein, the method comprises the following steps: in the gold nanorods solution that contains cetyl trimethyl ammonium bromide, under the condition that is 8.5-11 at pH value, add the alcoholic solution of esters of silicon acis to react, and carry out solid-liquid separation, obtain surface and be coated with the gold nanorods of meso-porous titanium dioxide silicon layer.

According to the preparation method of pharmaceutical carrier of the present invention, the method for preparing described gold nanorods aqueous solution can adopt the conventional method of this area to carry out.For example can adopt seed mediated growth method to prepare, its step is under the existence of cetyl trimethyl ammonium bromide, use sodium borohydride reduction gold chloride, obtain the seed solution of gold nano grain, this seed solution is added the monovalence gold ion growth from solution being obtained by ascorbic acid reduction gold chloride to obtain gold nanorods under cetyl trimethyl ammonium bromide exists.In addition, in seed growth, can be by regulating the amount of silver nitrate solution and sulfuric acid solution to control length and the draw ratio of gold nanorods.

According to the preparation method of pharmaceutical carrier of the present invention, in described gold nanorods aqueous solution, contain cetyl trimethyl ammonium bromide.Under preferable case, the concentration of the cetyl trimethyl ammonium bromide in described gold nanorods aqueous solution is 0.3-8 mM/l; More preferably the concentration of the cetyl trimethyl ammonium bromide in described gold nanorods aqueous solution is 0.5-1.2 mM/l.When the concentration of cetyl trimethyl ammonium bromide in the gold nanorods aqueous solution preparing by said method is during higher than above-mentioned preferred concentration, can obtain by centrifuge washing the gold nanorods aqueous solution of required cetyl trimethyl ammonium bromide concentration.The method of centrifuge washing can adopt method known in the field to carry out.

According to the preparation method of pharmaceutical carrier of the present invention, under preferable case, in the aqueous solution of described gold nanorods, take the gold nanorods concentration of gold element as 10 micromoles per liter ~ 2.5 mM/l; Consider from being easy to preparation, more preferably in described gold nanorods aqueous solution, take the gold nanorods concentration of gold element as 0.1 mM/l ~ 1 mM/l; Further in preferred described gold nanorods aqueous solution, take the gold nanorods concentration of gold element as 0.15 ~ 0.5 mM/l.

According to the preparation method of pharmaceutical carrier of the present invention, under normal circumstances, the length of described gold nanorods can be 5-200 nanometer, and under preferable case, described gold nanorods length is 10-150 nanometer; Consider from being conducive to cellular uptake, more preferably described gold nanorods length is 15-120 nanometer; Further preferred described gold nanorods length is 20-100 nanometer; Further preferred described gold nanorods length is 30-80 nanometer.

According to the preparation method of pharmaceutical carrier of the present invention, the consumption of described esters of silicon acis can be selected according to the amount of the gold nanorods in described gold nanorods aqueous solution.Under preferable case, respectively in gold element and element silicon, the gold nanorods in the aqueous solution of described gold nanorods and the mol ratio of described esters of silicon acis are 1:1-50; Consider from increasing silicon dioxide layer thickness and intensity, more preferably respectively in gold element and element silicon, the gold nanorods in the aqueous solution of described gold nanorods and the mol ratio of described esters of silicon acis are 1:2-10; Further, preferably respectively in gold element and element silicon, the gold nanorods in the aqueous solution of described gold nanorods and the mol ratio of described esters of silicon acis are 1:3-5.

According to the preparation method of pharmaceutical carrier of the present invention, under preferable case, in the described gold nanorods of gold element, with the mol ratio of cetyl trimethyl ammonium bromide be 1:1-20; Consider from thickness and the intensity of silicon dioxide layer, more preferably in the aqueous solution of the described gold nanorods of gold element, with the mol ratio of cetyl trimethyl ammonium bromide be 1:1.5-10; Further preferably in the aqueous solution of the described gold nanorods of gold element, with the mol ratio of cetyl trimethyl ammonium bromide be 1:2-4.

According to the preparation method of pharmaceutical carrier of the present invention, to the way of contact of described esters of silicon acis and gold nanorods aqueous solution, there is no particular limitation, for example, described esters of silicon acis can be joined in described gold nanorods aqueous solution.To the mode adding, also there is no particular limitation, can be disposable adding, and also can add in batches, considers when the uniformity from the meso-porous titanium dioxide silicon layer that obtains, preferably adds in batches.Consider from operability, preferably 2-5 time, the interval time more preferably adding for each time is identical.

According to the preparation method of pharmaceutical carrier of the present invention, under the condition that to contact at pH value be 8.5-11 of described esters of silicon acis and gold nanorods aqueous solution, carry out, under preferable case, under the condition that to contact at pH value be 9-10.5 of esters of silicon acis and gold nanorods aqueous solution, carry out.The adjusting of pH value can adopt alkaline pH value regulator to realize, and described alkaline pH value regulator can be the conventional alkali compounds in this area, for example, can use sodium hydroxide, potassium hydroxide etc.

According to the preparation method of pharmaceutical carrier of the present invention, described esters of silicon acis also comprises with the contact conditions of gold nanorods aqueous solution: the temperature contacting is 4-95 ℃, and the time of contact is 0.04-10 days; Under preferable case, the temperature of contact is 10-50 ℃, and the time of contact is 1-3 days.

According to the preparation method of pharmaceutical carrier of the present invention, described esters of silicon acis can be alkyl silicate; Preferred described alkyl is that carbon number is the alkyl of 1-4; The alkyl that more preferably carbon number is 1-3.In above-mentioned esters of silicon acis, preferably one or more in quanmethyl silicate, tetraethyl orthosilicate, silicic acid orthocarbonate and silicic acid four butyl esters.

According to the preparation method of pharmaceutical carrier of the present invention, described alcoholic solvent is as long as being easy to dissolve described esters of silicon acis and solvent soluble in water.For example can be less than 200 for methanol, ethanol or other molecular weight, be one or more of the alcohol apoplexy due to endogenous wind of liquid under normal temperature and pressure; Be preferably methanol and/ethanol.

According to the preparation method of pharmaceutical carrier of the present invention, the method comprises by centrifugalize, obtains surface and be coated with the gold nanorods of meso-porous titanium dioxide silicon layer.As long as the condition of described centrifugalize fully sedimentation surface is coated with the gold nanorods of meso-porous titanium dioxide silicon layer.Generally, described centrifugal speed can be 3000-15000 rev/min; The above-mentioned centrifugal time can be 5-100 minute.

The pharmaceutical carrier obtaining according to the preparation method of pharmaceutical carrier of the present invention, under preferable case, its maximum absorption band is in the scope of 620-1200 nanometer, more preferably in the scope of 650-1000 nanometer.

According to pharmaceutical composition of the present invention, to the type of the medicine of load on described carrier, there is no particular limitation, can carry out according to the actual needs the various types of medicines of load.For example can load hydrophilic and hydrophobic medicine.Under preferable case, described medicine is one or more in amycin, paclitaxel, vinblastine, cisplatin, camptothecine, DNA, siRNA and pharmaceutical grade protein.

According to pharmaceutical composition of the present invention, in preferred situation, take the weight of described carrier as benchmark, the carrying drug ratio of described pharmaceutical composition is 1-40 % by weight; More preferably take the weight of described carrier as benchmark, the carrying drug ratio of described pharmaceutical composition is 5-30 % by weight, and further, preferably take the weight of described carrier as benchmark, the carrying drug ratio of described pharmaceutical composition is 10-20 % by weight.

The computing formula of carrying drug ratio is:

Weight × 100% of carrying drug ratio (% by weight)=(weight of medicine on carrier)/carrier.

In the present invention, to by drug loading the method on above-mentioned carrier there is no particular limitation, as long as can by a certain amount of drug loading to the method on carrier all can, can adopt known the whole bag of tricks.It is for example, described that by drug loading, the method on carrier can be that drug solution is contacted with described pharmaceutical carrier.

In the method that drug solution is contacted with described pharmaceutical carrier, the consumption of described drug solution can be selected according to the amount of described pharmaceutical carrier.Particularly, the weight ratio of the medicine in described pharmaceutical carrier and described drug solution can be 1:0.1-10; Be preferably 1:0.5-5.In addition, to the concentration of described drug solution, there is no particular limitation, but be preferably 0.01-5 mg/ml; More preferably 0.05-1.5 mg/ml.

In addition, drug solution is comprised with the condition that described pharmaceutical carrier contacts: the temperature of contact is 1-50 ℃, the time of contact is 1-4 days.Preferred described contact is under agitation carried out.

In the method that drug solution is contacted with described pharmaceutical carrier, to the type of the medicine in described drug solution, there is no particular limitation, can select according to the actual needs.It can be for example the medicine of hydrophilic or hydrophobicity type.Under preferable case, described medicine is one or more in amycin, paclitaxel, vinblastine, cisplatin, camptothecine, DNA and siRNA and pharmaceutical grade protein.

According to pharmaceutical composition of the present invention, it comprises the carrier of said structure of the present invention, because this carrier can bio-imaging, and can be for the diagnosis of various diseases; In addition, because this carrier can absorb the laser of near infrared region, and launch fluorescence and part light energy conversion is become to heat, therefore, can be by the release of near-infrared laser control medicine, thereby for the treatment of various diseases, for example can be for the chemotherapy of the diseases such as cancer.

As mentioned above, pharmaceutical composition of the present invention comprises the carrier of said structure of the present invention, because this carrier can bio-imaging, and can be for the diagnosis of various diseases; In addition, because this carrier can absorb the laser of near infrared region, and launch fluorescence and part light energy conversion is become to heat, therefore, can be by the release of near-infrared laser control medicine, thereby for the treatment of various diseases.Particularly can use it for diagnosis and/or treatment tumor.While using it for diagnosis and treatment tumor, can realize the diagnosis of tumor simultaneously, and chemotherapy and two kinds of therapeutic modalities of thermotherapy.

There is no special requirement as the kind of tumor, as long as according to the applicable medicine of the type load of tumor.Such as the tumor of applicable diagnosis and treatment comprises: breast carcinoma, pulmonary carcinoma, melanoma, hepatocarcinoma, skin carcinoma, cervical cancer, bladder cancer, cancer of pancreas, gastric cancer etc.

Below will describe the present invention by embodiment, but the present invention is not limited in following embodiment.

In following examples, gold chloride, cetyl trimethyl ammonium bromide, silver nitrate, ascorbic acid, sodium borohydride, tetraethyl orthosilicate are all purchased from Chemical Reagent Co., Ltd., Sinopharm Group.Doxorubicin hydrochloride (DOX) is purchased from Jinan Wei Dou medical material company limited.If no special instructions, in the present invention, all waters are redistilled water.

In following examples, the aperture of the rod length of gold nanorods, gold nanorods draw ratio, meso-porous titanium dioxide silicon layer thickness, meso-porous titanium dioxide silicon layer adopts transmission electron microscope (being purchased from FEI Co. of the U.S., Tecnai G2F20S-TWIN model) to measure.

Mesoporous silicon oxide gold-covered nano rod carrier is after lyophilizing, and the full-automatic specific surface area analysis instrument of Tristar II3020 type that uses Micromeritics Instrument Corp. U.S.A to produce is measured its specific surface area, aperture and pore volume.In following examples, the multi-functional microplate reader of Tecan Infinite M200 type continuous spectrum that maximum absorption wavelength uses FEI Co. of the U.S. to produce is measured.

Embodiment 1

The present embodiment is for illustrating the preparation of pharmaceutical composition.

1) preparation of pharmaceutical carrier

In the beaker of 20 milliliters, be that 0.1 mol/L is mixed with the aqueous solution of chloraurate (concentration is 0.01 mol/L) of 250 microlitres by the concentration of the cetyl trimethyl ammonium bromide aqueous solution of 7.5 milliliters, adding water to cumulative volume is 9.4 milliliters; Then add the sodium borohydride aqueous solution of 0.6 milliliter (concentration is 0.01 mol/L) of the cooling mistake of frozen water; At 25 ℃, stir 3 hours, obtain gold nano grain seed solution.

In the flask of 150 milliliters, add the cetyl trimethyl ammonium bromide (concentration is 0.1 mol/L) of 100 milliliters, the aqueous solution of chloraurate (concentration is 0.01 mol/L) of 5 milliliters, the silver nitrate aqueous solution (concentration is 0.01 mol/L) of 0.8 milliliter, the aqueous sulfuric acid (concentration is 0.5 mol/L) of 2 milliliters, and the aqueous ascorbic acid of 800 microlitres (concentration is 0.1 mol/L); Then add the gold nano grain seed solution obtained above of 240 microlitres, reaction is carried out 24 hours at 30 ℃, obtains gold nanorods aqueous solution.As shown in Figure 1, the rod of gold nanorods is grown up and is about 50 nanometers, and gold nanorods draw ratio is 3.6, and maximum absorption wave wavelength is 759 nanometers.

Then,, by the gold nanorods solution centrifugal washing of 40 milliliters of above-mentioned preparations 2 times, all making residual volume for twice is 2.53 milliliters.After centrifugal completing, adding water to for the first time 40 milliliters disperses again, after centrifugal completing, adding water to for the second time 20 milliliters disperses again, in the gold nanorods aqueous solution finally obtaining, cetyl trimethyl ammonium bromide concentration is 0.8 mM/l, and take the gold nanorods concentration of gold element as 0.25 mM/l.Then, under agitation adding the sodium hydrate aqueous solution adjusting pH value of 0.1 mol/L was 10, then added the ethyl orthosilicate methanol solution (concentration is 20 % by weight) of 60 microlitres every 30 minutes, altogether added three times.Reaction is under agitation carried out 2 days at 27 ℃, obtain the gold nanorods that surface is coated with meso-porous titanium dioxide silicon layer (be pharmaceutical carrier, below also referred to as Au SiO ₂) solution, then water carries out centrifuge washing 2 times to remove the cetyl trimethyl ammonium bromide in solution, obtains the Au@SiO after centrifuge washing ₂, by Au@SiO ₂carry out lyophilizing and obtain 9.5 milligrams of dried Au@SiO ₂.As shown in Figure 2, meso-porous titanium dioxide silicon layer thickness is 30 nanometers, and the aperture of meso-porous titanium dioxide silicon layer is 3-4 nanometer; In addition the Au@SiO obtaining, ₂specific surface area be 80 meters squared per gram, pore volume is 0.8 cubic centimetre/gram; The maximum absorption wavelength of the pharmaceutical carrier obtaining in addition, is 770 nanometers.

2) preparation of pharmaceutical composition

In beaker, by 1.6 milligrams of dried Au@SiO ₂be scattered in the amycin aqueous solution (concentration of amycin is 0.1 mg/ml) of 4 milliliters, use aluminium-foil paper lucifuge, and at 25 ℃, stir 4 days, obtain the solution that contains pharmaceutical composition, then obtain pharmaceutical composition (below also referred to as Au@SiO through centrifugal drying ₂-DOX).The carrying drug ratio of this pharmaceutical composition is 14.5 % by weight.

Above-mentioned carrying drug ratio adopts following methods to measure: before above-mentioned centrifugal drying, medicine carrying solution centrifugal takes a morsel, in measurement supernatant, amycin is in the characteristic absorption of 480 nanometers, by concentration-absorption standard curve of comparison amycin, the amount of carrying medicament not in the solution that obtains containing pharmaceutical composition.And calculate carrying drug ratio by following formula:

Weight × 100% of carrying drug ratio (% by weight)=(dropping into the weight-not weight of carrying medicament of medicine)/carrier.

The maximum absorption wavelength of the pharmaceutical composition obtaining in addition, is 790 nanometers.

Embodiment 2

1) preparation of pharmaceutical carrier

In the beaker of 20 milliliters, the cetyl trimethyl ammonium bromide aqueous solution of 7.5 milliliters (concentration of cetyl trimethyl ammonium bromide is 0.1 mol/L) is mixed with the aqueous solution of chloraurate (concentration is 0.01 mol/L) of 250 microlitres, and adding water to cumulative volume is 9.4 milliliters; Then add the sodium borohydride aqueous solution of 0.6 milliliter (concentration is 0.01 mol/L) of the cooling mistake of frozen water; At 25 ℃, stir 3 hours, obtain gold nano grain seed solution.

In the flask of 150 milliliters, add the cetyl trimethyl ammonium bromide (concentration is 0.1 mol/L) of 100 milliliters, the aqueous solution of chloraurate (concentration is 0.01 mol/L) of 5 milliliters, the silver nitrate aqueous solution (concentration is 0.01 mol/L) of 2 milliliters, the aqueous sulfuric acid (concentration is 0.5 mol/L) of 4 milliliters, and the aqueous ascorbic acid of 800 microlitres (concentration is 0.1 mol/L); Then add the gold nano grain seed solution gold nano grain obtained above of 240 microlitres, reaction is carried out 24 hours at 30 ℃, obtains gold nanorods aqueous solution.Known by transmission electron micrograph, the rod length of gold nanorods is 60 nanometers, and gold nanorods draw ratio is 4; In addition, the maximum absorption wave wavelength of gold nanorods is 800 nanometers.

Then,, by the gold nanorods solution centrifugal washing of 40 milliliters of above-mentioned preparations 2 times, all making residual volume for twice is 2.83 milliliters.After centrifugal completing, adding water to for the first time 40 milliliters disperses again, after centrifugal completing, adding water to for the second time 20 milliliters disperses again, in the gold nanorods aqueous solution finally obtaining, cetyl trimethyl ammonium bromide concentration is 1 mM/l, and take the gold nanorods concentration of gold element as 0.5 mM/l.Then, under agitation adding the sodium hydrate aqueous solution adjusting pH value of 0.1 mol/L was 9.0, then added the ethyl orthosilicate methanol solution (concentration is 20 % by weight) of 70 microlitres every 30 minutes, altogether added three times.Reaction is under agitation carried out 2 days at 35 ℃, obtain the gold nanorods that surface is coated with meso-porous titanium dioxide silicon layer (be pharmaceutical carrier, below also referred to as Au SiO ₂) solution, then water carries out centrifuge washing 2 times to remove the cetyl trimethyl ammonium bromide in solution, obtains the Au@SiO after centrifuge washing ₂, by Au@SiO ₂carry out lyophilizing and obtain 13.2 milligrams of dried Au@SiO ₂.The thickness of meso-porous titanium dioxide silicon layer is 40 nanometers, and the aperture of meso-porous titanium dioxide silicon layer is 5-7 nanometer; In addition the Au@SiO obtaining, ₂specific surface area be 95 meters squared per gram, pore volume is 1.05 cubic centimetres/gram.The maximum absorption wave wavelength of the pharmaceutical carrier obtaining is 820 nanometers.

2) preparation of pharmaceutical composition

In beaker, by 2.4 milligrams of dried Au@SiO ₂be scattered in the paclitaxel aqueous solution (concentration of paclitaxel is 0.2 mg/ml) of 4 milliliters, use aluminium-foil paper lucifuge, and stir 3 days at 25 ℃, obtain the solution that contains pharmaceutical composition, then obtain pharmaceutical composition through centrifugal drying.The carrying drug ratio of this pharmaceutical composition is 17 % by weight.

Above-mentioned carrying drug ratio adopts following methods to measure: before above-mentioned centrifugal drying, the medicine carrying solution centrifugal that takes a morsel, uses high performance liquid chromatograph to measure the amount of paclitaxel in supernatant, the amount of carrying medicament not in the solution that obtains containing pharmaceutical composition.And calculate carrying drug ratio by following formula,

Weight × 100% of carrying drug ratio (% by weight)=(dropping into the amount-not amount of carrying medicament of medicine)/carrier.

The maximum absorption wavelength of the pharmaceutical composition obtaining in addition, is 840 nanometers.

Embodiment 3

1) preparation of pharmaceutical carrier

In the flask of 150 milliliters, add the cetyl trimethyl ammonium bromide (concentration is 0.1 mol/L) of 100 milliliters, the aqueous solution of chloraurate (concentration is 0.01 mol/L) of 5 milliliters, the silver nitrate aqueous solution (concentration is 0.01 mol/L) of 2 milliliters, the aqueous sulfuric acid (concentration is 0.5 mol/L) of 4 milliliters, and the aqueous ascorbic acid of 800 microlitres (concentration is 0.1 mol/L); Then add the gold nano grain seed solution gold nano grain obtained above of 240 microlitres, reaction is carried out 24 hours at 30 ℃, obtains gold nanorods aqueous solution.Known by transmission electron micrograph, the rod length of gold nanorods is 70 nanometers, and gold nanorods draw ratio is 4.2; In addition, the maximum absorption wave wavelength of gold nanorods is 850 nanometers.

Then,, by the gold nanorods solution centrifugal washing of 40 milliliters of above-mentioned preparations 2 times, all making residual volume for twice is 2.19 milliliters.After centrifugal completing, adding water to for the first time 40 milliliters disperses again, after centrifugal completing, adding water to for the second time 20 milliliters disperses again, in the gold nanorods aqueous solution finally obtaining, cetyl trimethyl ammonium bromide concentration is 0.6 mM/l, and take the gold nanorods concentration of gold element as 0.5 mM/l.Then, under agitation adding the sodium hydrate aqueous solution adjusting pH value of 0.1 mol/L was 10.5, then added the ethyl orthosilicate methanol solution (concentration is 20 % by weight) of 50 microlitres every 30 minutes, altogether added three times.Reaction is under agitation carried out 1.5 days at 26 ℃, obtain the gold nanorods that surface is coated with meso-porous titanium dioxide silicon layer (be pharmaceutical carrier, below also referred to as Au SiO ₂) solution, then water carries out centrifuge washing 2 times to remove the cetyl trimethyl ammonium bromide in solution, obtains the Au@SiO after centrifuge washing ₂, by Au@SiO ₂carry out lyophilizing and obtain 14.5 milligrams of dried Au@SiO ₂.Transmission electron microscope Fig. 3 by pharmaceutical carrier is known, and the thickness of meso-porous titanium dioxide silicon layer is 15 nanometers, and the aperture of meso-porous titanium dioxide silicon layer is 2-5 nanometer; In addition the Au@SiO obtaining, ₂specific surface area be 65 meters squared per gram, pore volume is 0.65 cubic centimetre/gram.The maximum absorption wave wavelength of the pharmaceutical carrier obtaining is 870 nanometers.

2) preparation of pharmaceutical composition

In beaker, by 3.0 milligrams of dried Au@SiO ₂be scattered in the siRNA aqueous solution (concentration is 1.0 mg/ml) of 4 milliliters, use aluminium-foil paper lucifuge, and stir 2 days at 25 ℃, obtain the solution that contains pharmaceutical composition, then obtain pharmaceutical composition through centrifugal drying.The carrying drug ratio of this pharmaceutical composition is 18 % by weight.

Above-mentioned carrying drug ratio adopts following methods to measure: before above-mentioned centrifugal drying, the medicine carrying solution centrifugal that takes a morsel, uses HPLC to measure the amount of siRNA in supernatant, the amount of carrying medicament not in the solution that obtains containing pharmaceutical composition.And calculate carrying drug ratio by following formula,

The maximum absorption wave wavelength of the pharmaceutical composition obtaining in addition, is 890 nanometers.

Embodiment 4

1) preparation of pharmaceutical carrier

Preparation method according to gold nanorods in embodiment 1 is carried out, different is to use the silver nitrate aqueous solution (concentration is 0.01 mol/L) of 2.5 milliliters, the aqueous sulfuric acid (concentration is 0.5 mol/L) of 5 milliliters, the rod length of similarly preparing gold nanorods is 80 nanometers, draw ratio is 4.5 gold nanorods solution, wherein contain the cetyl trimethyl ammonium bromide of 0.09 mol/L, its maximum absorption wavelength is 870 nanometers.

Then,, by the gold nanorods solution centrifugal washing of 40 milliliters of above-mentioned preparations 2 times, all making residual volume for twice is 3.26 milliliters.After centrifugal completing, adding water to for the first time 40 milliliters disperses again, after centrifugal completing, adding water to for the second time 20 milliliters disperses again, in the gold nanorods aqueous solution finally obtaining, cetyl trimethyl ammonium bromide concentration is 1.2 mM/ls, and take the gold nanorods concentration of gold element as 0.5 mM/l.Then, under agitation adding the sodium hydrate aqueous solution adjusting pH value of 0.1 mol/L was 11, then added the methanol solution (concentration is 20 % by weight) of the quanmethyl silicate of 80 microlitres every 30 minutes, altogether added three times.Reaction is under agitation carried out 1 day at 45 ℃, obtain the gold nanorods that surface is coated with meso-porous titanium dioxide silicon layer (be pharmaceutical carrier, below also referred to as Au SiO ₂) solution, then water carries out centrifuge washing 2 times to remove the cetyl trimethyl ammonium bromide in solution, obtains the Au@SiO after centrifuge washing ₂, by Au@SiO ₂carry out lyophilizing and obtain 17 milligrams of dried Au@SiO ₂.Transmission electron microscope figure by pharmaceutical carrier is known, and the thickness of meso-porous titanium dioxide silicon layer is 25 nanometers, and the aperture of meso-porous titanium dioxide silicon layer is 3-4 nanometer; In addition the Au@SiO obtaining, ₂surface area be 85 meters squared per gram, pore volume is 1.2 cubic centimetres/gram.The maximum absorption wave wavelength of the pharmaceutical carrier obtaining is 885 nanometers.

2) preparation of pharmaceutical composition

In beaker, by 3 milligrams of dried Au@SiO ₂be scattered in the dimethyl sulphoxide solution (concentration of cisplatin is 0.5 mg/ml) of the cisplatin of 4 milliliters, use aluminium-foil paper lucifuge, and stir 2 days at 15 ℃, obtain the solution that contains pharmaceutical composition, then obtain pharmaceutical composition through centrifugal drying.The carrying drug ratio of this pharmaceutical composition is 12 % by weight.

Above-mentioned carrying drug ratio adopts following methods to measure: before above-mentioned centrifugal drying, the medicine carrying solution centrifugal that takes a morsel, measures the content of cisplatin in supernatant by high pressure liquid chromatography, the amount of carrying medicament not in the solution that obtains containing pharmaceutical composition.And calculate carrying drug ratio by following formula,

The maximum absorption wave wavelength of the pharmaceutical composition obtaining in addition, is 895 nanometers.

Embodiment 5

The present embodiment is for illustrating the photo-thermal transition effects of pharmaceutical carrier of the present invention

The pharmaceutical carrier of preparation in embodiment 1 is scattered in the water of 20 milliliters, gets 200 microlitres and put into 96 orifice plates.Using spot diameter is 2 millimeters, and power is 150 milliwatts or 250 milliwatts, and wavelength is the laser irradiation of 780 nanometers.By the temperature of temperature sensitive thermometer recording solution, and draw out temperature-irradiation time curve (Fig. 4).As shown in Figure 4, under 150 milliwatt laser irradiations, the highest 38 degree that rise to of the temperature of nano-particle solution, and under 250 milliwatt laser irradiations, the highest 48 degree that rise to.

Embodiment 6

The present embodiment is for illustrating the release of pharmaceutical composition of the present invention.

Get the pharmaceutical composition of preparation in embodiment 1, divide two groups and carry out release experiment.

First group: the pharmaceutical composition 3 parts (0.5 milligram every part) of preparation in embodiment 1 is used respectively to pH=7.4,6.0,1.0 milliliters of dispersions of phosphate buffered solution of 4.5, be placed in 37 ℃ of shaking tables and start release experiment.Took out once every 1 hour, centrifugal, collect supernatant, and by the characteristic absorption of 480 nanometers, the amycin quality that measuring and calculating has discharged.Residue disperses with identical buffer solution, puts back to shaking table, proceeds drug release experiment.Release experiment is carried out 12 hours altogether.After experiment finishes, calculate release amount hourly and account for the ratio of total pharmaceutical pack carrying capacity, draw release curve according to cumulative release rate-drug release time, specifically as shown in Figure 5.

Second group: the pharmaceutical composition 3 parts (0.5 milligram every part) of preparation in embodiment 1 is used respectively to pH=7.4,6.0,1.0 milliliters of dispersions of phosphate buffered solution of 4.5, with wavelength be 760 nanometers, power is 250 milliwatts, spot diameter is the laser irradiation of 2 millimeters, make solution temperature rise to 48 ℃, carry out release experiment.Took out once every 1 hour, centrifugal, collect supernatant, and by the characteristic absorption of 480 nanometers, the amycin quality that measuring and calculating has discharged.Residue disperses with identical buffer solution, again uses laser irradiation, proceeds drug release experiment.Release experiment is carried out 12 hours altogether.After experiment finishes, calculate release amount hourly and account for the ratio of total pharmaceutical pack carrying capacity, draw release curve according to cumulative release rate-drug release time, specifically as shown in Figure 5.

As shown in Figure 5, in the time of 37 ℃, drug releasing rate is very slow, although slightly faster than drug release rate under neutrallty condition at low pH, and no significant difference.But it is known to contrast first group of release experiment, and after laser irradiation raises solution temperature, drug release rate is accelerated greatly; PH is lower, and it is more that drug release rate is accelerated, and therefore, pharmaceutical composition of the present invention can be by the speed of external its release of laser irradiation control, and can be in response to the distinctive weak acid environment in tumor tissues inside.

Embodiment 7

Get the pharmaceutical composition of preparation in embodiment 2, divide two groups and carry out release experiment.

First group: the pharmaceutical composition 3 parts (0.5 milligram every part) of preparation in embodiment 2 is used respectively to pH=7.4,6.0,1.0 milliliters of dispersions of phosphate buffered solution of 4.5, be placed in 37 ℃ of shaking tables and start release experiment.Took out once every 1 hour, centrifugal, collect supernatant, and use high performance liquid chromatograph, the quality of the paclitaxel that measuring and calculating has discharged.Residue disperses with identical buffer solution, puts back to shaking table, proceeds drug release experiment.Release experiment is carried out 12 hours altogether.After experiment finishes, calculate release amount hourly and account for the ratio of total pharmaceutical pack carrying capacity, draw release curve according to cumulative release rate-drug release time, specifically as shown in Figure 6.

Second group: the pharmaceutical composition 3 parts (0.5 milligram every part) of preparation in embodiment 2 is used respectively to pH=7.4,6.0,1.0 milliliters of dispersions of phosphate buffered solution of 4.5, with wavelength be 840 nanometers, power is 250 milliwatts, spot diameter is the laser irradiation of 2 millimeters, make solution temperature rise to 48 ℃, carry out release experiment.Took out once every 1 hour, centrifugal, collect supernatant, and use high performance liquid chromatograph to calculate the paclitaxel quality having discharged.Residue disperses with identical buffer solution, again uses laser irradiation, proceeds drug release experiment.Release experiment is carried out 12 hours altogether.After experiment finishes, calculate release amount hourly and account for the ratio of total pharmaceutical pack carrying capacity, draw release curve according to cumulative release rate-drug release time, specifically as shown in Figure 6.

As shown in Figure 6, in the time of 37 ℃, drug releasing rate is very slow, although slightly faster than drug release rate under neutrallty condition at low pH, and no significant difference.But it is known to contrast first group of release experiment, and after laser irradiation raises solution temperature, drug release rate is accelerated greatly; PH is lower, and it is more that drug release rate is accelerated, and therefore, pharmaceutical composition of the present invention can be by the speed of external its release of laser irradiation control, and can be in response to the distinctive weak acid environment in tumor tissues inside.

Embodiment 8

The present embodiment is used for illustrating that pharmaceutical carrier of the present invention and pharmaceutical composition are for cancerous cell imaging.

1) cultivation of people's lung cancer A549 cell

People's lung cancer A549 cell is inoculated in 35 millimeters of co-focusing imaging culture dishs, use DMEM fluid medium (streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls) in 37 ℃, in 5% CO2 gas incubator, cultivate phosphate buffered solution washed twice 24 hours.Then add respectively the Au@SiO that contains 5 micromoles per liter equivalence amycin ₂prepared by-DOX(embodiment 1) and contain Au@SiO ₂(prepared by embodiment 1, the Au@SiO of its consumption and 5 micromoles per liter equivalence amycin ₂contained Au@SiO in-DOX ₂identical) complete medium (DMEM fluid medium, its streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls), and process respectively cell 12 hours.

2) in lysosome, locate altogether

Abandon cell culture medium, add containing 100 nanomoles/liter the serum-free DMEM of lysosome dyestuff (LysoTracker Green) cultivate based on 37 ℃ of incubated cells 45 minutes.With phosphate buffered solution washed cell three times, add serum-free DMEM fluid medium, micro-Microscopic observation.Single photon mode imaging: 488 nanometer lasers excite, collecting 530 nano fluorescent signals is organelle locating information, and collects the red fluorescence signal of DOX in 580-610 nanochannel region original position.Two-photon mode imaging: excitation source is adjusted to two-photon pattern, and 780 nanometer femtosecond lasers (fs-pulse, Maitai of Spectra Physics in USA), collect 530 nanometer utilizing emitted light signals.Thus, obtain constitutional diagram 7.Wherein, A in Fig. 7 is amycin (DOX) and lysosome (Lyso) network for location altogether, same intracellular amycin (DOX, represent by redness) photo and lysosome (Lyso, represent by green) after photo-overlap, can observe yellow speckle, show that intracellular amycin is positioned in lysosome; B in Fig. 7 is Au@SiO ₂carrier and lysosome (Lyso) be network for location altogether, same intracellular Au@SiO ₂after carrier (representing by blueness) photo and lysosome (Lyso, with green represent) photo-overlap, can observe glaucous speckle, show intracellular Au@SiO ₂be positioned in lysosome; C in Fig. 7 is Au@SiO ₂with amycin (DOX) network for location altogether, same intracellular Au@SiO ₂after carrier (representing by blueness) photo and amycin (DOX, with redness represent) photo-overlap, can observe peach speckle, show intracellular Au@SiO ₂with amycin in cell in same site; D in Fig. 7 is Au@SiO ₂, amycin (DOX) and lysosome (Lyso) three network for location altogether, same intracellular Au@SiO ₂after carrier (representing by blueness) photo, amycin (DOX, with redness represent) photo and lysosome (Lyso, with green represent) are overlapping, can observe white speckle, show intracellular Au@SiO ₂be positioned in lysosome together with amycin.As seen from Figure 7, by nano-carrier Au@SiO ₂two-photon itself causes fluorescent effect and determines after it enters A549 cell and can accumulate in lysosome (Lyso).

Embodiment 9

1) cultivation of people's lung cancer A549 cell

2) in mitochondrion, locate altogether

Abandon cell culture medium, add containing 100 nanomoles/liter 37 ℃ of incubated cells of serum-free DMEM culture medium 45 minutes of mitochondrion dyestuff (MitoTracker Green).With phosphate buffered solution washed cell three times, add serum-free DMEM fluid medium, micro-Microscopic observation.Single photon mode imaging: 488 nanometer lasers excite, collecting 530 nano fluorescent signals is organelle locating information, and collects the red fluorescence signal of amycin in 580-610 nanochannel region original position.Two-photon mode imaging: excitation source is adjusted to two-photon pattern, and 780 nanometer femtosecond lasers (fs-pulse, Maitai of Spectra Physics in USA), collect 530 nanometer utilizing emitted light signals.Thus, obtain constitutional diagram 8.Wherein, A in Fig. 8 is amycin (DOX) and mitochondrion (Mito) network for location altogether, same intracellular amycin (DOX, represent by redness) photo and mitochondrion (Mito, represent by green) after photo-overlap, do not observe yellow speckle, show the indefinite mitochondrion that is positioned at of intracellular amycin; B in Fig. 8 is Au@SiO ₂carrier and mitochondrion (Mito) be network for location altogether, same intracellular Au@SiO ₂after carrier (representing by blueness) photo and mitochondrion (Mito, with green represent) photo-overlap, only observe a small amount of glaucous speckle, show in cell, to only have a small amount of Au@SiO ₂be positioned in mitochondrion; C in Fig. 8 is Au@SiO ₂with amycin (DOX) network for location altogether, same intracellular Au@SiO ₂after carrier (representing by blueness) photo and amycin (DOX, with redness represent) photo-overlap, can observe peach speckle, show intracellular Au@SiO ₂with amycin in cell in same site; D in Fig. 8 is Au@SiO ₂, amycin (DOX) and mitochondrion (Mito) three be network for location altogether, same intracellular Au@SiO ₂after carrier (representing by blueness) photo, amycin (DOX, with redness represent) photo and mitochondrion (Mito, with green represent) are overlapping, can observe obvious pink speckle and minute quantity white dot, show intracellular Au@SiO ₂in same site, and only there is minute quantity Au@SiO with amycin ₂be positioned in mitochondrion together with amycin.As seen from Figure 8, by nano-carrier Au@SiO ₂two-photon effect itself can be determined that it enters after A549 cell and have and accumulate on a small quantity in mitochondrion.

Embodiment 10

Lung carcinoma cell in embodiment 8 is replaced by human breast cancer cell MCF-7, other steps are identical, obtain constitutional diagram 9, wherein, the A in Fig. 9 is amycin (DOX) and lysosome (Lyso) network for location altogether, same intracellular amycin (DOX, represent by redness) photo and lysosome (Lyso, represent by green) after photo-overlap, can observe yellow speckle, show that intracellular amycin is positioned in lysosome; B in Fig. 9 is Au@SiO ₂carrier and lysosome (Lyso) be network for location altogether, same intracellular Au@SiO ₂after carrier (representing by blueness) photo and lysosome (Lyso, with green represent) photo-overlap, can observe glaucous speckle, show intracellular Au@SiO ₂be positioned in lysosome; C in Fig. 9 is Au@SiO ₂with amycin (DOX) network for location altogether, same intracellular Au@SiO ₂after carrier (representing by blueness) photo and amycin (DOX, with redness represent) photo-overlap, can observe peach speckle, show intracellular Au@SiO ₂with amycin in cell in same site; D in Fig. 9 is Au@SiO ₂, amycin (DOX) and lysosome three be network for location altogether, same intracellular Au@SiO ₂after carrier (representing by blueness) photo, amycin (DOX, with redness represent) photo and lysosome (Lyso, with green represent) are overlapping, can observe white speckle, show intracellular Au@SiO ₂be positioned in lysosome together with amycin.As seen from Figure 9, by nano-carrier Au@SiO ₂two-photon effect itself can be determined after it enters human breast cancer cell MCF-7 cell and can be enriched in lysosome.

Embodiment 11

Lung carcinoma cell in embodiment 8 is replaced by human breast cancer cell MCF-7, other steps are identical, obtain combining Figure 10, wherein, the A in Figure 10 is amycin (DOX) and mitochondrion (Mito) network for location altogether, same intracellular amycin (DOX, represent by redness) photo and mitochondrion (Mito, represent by green) after photo-overlap, only observe yellow speckle, show that intracellular amycin only has to enter on a small quantity in mitochondrion; B in Figure 10 is Au@SiO ₂carrier and lysosome (Mito) be network for location altogether, same intracellular Au@SiO ₂after carrier (representing by blueness) photo and mitochondrion (Mito, with green represent) photo-overlap, only observe a small amount of glaucous speckle, show in cell, to only have a small amount of Au@SiO ₂be positioned in mitochondrion; C in Figure 10 is Au@SiO ₂with amycin (DOX) network for location altogether, same intracellular Au@SiO ₂after carrier (representing by blueness) photo and amycin (DOX, with redness represent) photo-overlap, can observe peach speckle, show intracellular Au@SiO ₂with amycin in cell in same site; D in Figure 10 is Au@SiO ₂, amycin (DOX) and mitochondrion (Mito) three be network for location altogether, same intracellular Au@SiO ₂after carrier (representing by blueness) photo, amycin (DOX, with redness represent) photo and mitochondrion (Mito, with green represent) are overlapping, can observe obvious pink speckle and minute quantity white dot, show intracellular Au@SiO ₂in same site, and only there is minute quantity Au@SiO with amycin ₂be positioned in mitochondrion together with amycin.As seen from Figure 10, by nano-carrier Au@SiO ₂two-photon effect itself can be determined that it enters after A549 cell and have and be enriched on a small quantity in mitochondrion.

By above-described embodiment, 8-11 is known, and because gold nanorods can be excited by the near-infrared femtosecond pulse of 780 nanometers, emitting fluorescence signal is in 450-600 nanometer, but do not excited by single photon 488 nanometer lasers, can distinguish the signal of gold nanorods and dyestuff.And can utilize software to locate altogether organelle, DOX and gold nanorods.

Embodiment 12

The present embodiment is for illustrating the absorption of cancerous cell to pharmaceutical composition of the present invention.

1) cultivation of people's lung cancer A549 cell

People's lung cancer A549 cell is inoculated in 35 millimeters of co-focusing imaging culture dishs, use DMEM fluid medium (to contain 10 % by weight hyclones, the streptomycin of 1 % by weight penicillin and 100 units per ml, the glutamine of 2 mM/ls) in 37 ℃, in 5% CO2 gas incubator, cultivate 24 hours.

2) cellular uptake quantitative determination

Cultured cells in step 1 is inoculated in 6 orifice plates, 80000 cells/well, add 2 milliliters of complete medium (DMEM fluid mediums, its streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls), be placed in 37 ℃, in 5% CO2 gas incubator, cultivate 24 hours.Then in the DMEM fluid medium of new preparation, add respectively the DOX of 5 micromoles per liter and the Au@SiO of 5 micromoles per liter equivalence DOX concentration ₂prepared by-DOX(embodiment 1), mix and respectively get 2 milliliters and add in 6 orifice plates afterwards, every kind of concentration arranges 4 parallel holes, 37 ℃, in 5% CO2 gas incubator, cultivates.Use respectively trypsin digestion and cell in different time points (3,6,12,24 hours), 800g is centrifugal, and then 0.5 milliliter of resuspended preparation cell suspension of hanks buffer detect fluorescent value in cell with flow cytometer.Detected parameters: take fixing streaming laser voltage in experimentation, 488 nanometer lasers excite, collects FL2 access detection signal.Gathering the mean fluorecence value of 10,000 cells, by several Duplicate Samples calculating mean values, is this cellular uptake amycin (DOX) of this time point and Au@SiO ₂the fluorescent quantitation value of-DOX.Get fluorescent quantitation value, the mapping of Origin software, analysis of cells is taken in the trend of amycin (DOX), as Figure 11.

As seen in Figure 11 with respect to independent use anticancer drugs, doxorubicin, with carrier A u@SiO of the present invention ₂bag carries amycin can increase cellular uptake amount about 5 times.

Embodiment 13

The present embodiment is for illustrating pharmaceutical carrier of the present invention and the impact of pharmaceutical composition on cell viability.

1) cell culture

People's lung cancer A549 cell is incubated at respectively to complete medium (DMEM fluid medium, its streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls) in, be placed in 37 ℃, in 5% CO2 gas incubator, cultivate 24 hours.

2) cell viability is measured

With 6000 cells/well density, cell is inoculated in 96 orifice plates.After adherent 24 hours, adding respectively containing concentration is amycin and the Au@SiO of 0.1,1.0,5.0,10 micromoles per liter ₂prepared by-DOX(embodiment 1, concentration is in the amycin of load) complete medium (DMEM fluid medium, its streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls).Processing after 24 hours, 48 hours, 72 hours, remove culture medium; Every hole adds 110 microlitres, containing 10%(volume ratio) cell culture fluid of CCK-8, in incubator, hatch after 2 hours; In microplate reader, be determined at 450 nanometers absorbances, 600 nanometers light absorption values are reference wavelength.After every group of light absorption value deduction blank solution light absorption value, every hole respective value divided by matched group as cell viability.Every group arranges 6 parallel holes, and every group of experiment in triplicate.Cell viability measurement result as shown in Figure 12 and Figure 13.Figure 12 is the graph of a relation of doxorubicin concentration and cell viability; Figure 13 is Au@SiO ₂the graph of a relation of-DOX concentration (concentration is in the amycin of load) and cell viability.Contrast Figure 12 and Figure 13, can find out, with respect to the isocyatic anticancer drugs, doxorubicin of independent use, and carrier A u SiO of the present invention ₂there is the effect that slows down amycin reduction cell viability, the effect that amycin slowly discharges from carrier is described.This feature illustrates that this carrier is specially adapted to require the treatment needs of medicament slow release.

Embodiment 14

The present embodiment is for illustrating the cell viability after laser irradiation.

1) cell culture

People's lung cancer A549 cell, with 3000 cells/well density, is inoculated in 96 orifice plates.Use DMEM fluid medium (streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls) in 37 ℃, in 5% CO2 gas incubator, after adherent 24 hours, add respectively the Au@SiO that contains 5 micromoles per liter equivalence amycin ₂prepared by-DOX(embodiment 1) and contain Au@SiO ₂(prepared by embodiment 1, the Au@SiO of its consumption and 5 micromoles per liter equivalence amycin ₂contained Au@SiO in-DOX ₂identical) complete medium (complete medium is DMEM fluid medium, its streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls), and process respectively cell 12 hours.

2) cell survival rate is measured

Before laser irradiation, phosphate buffered solution washing 2 times for cell, is that 780 nanometers, power are the laser irradiation 8 minutes of 24 watts/square centimeter with wavelength.After irradiation, continue to cultivate 12 hours, 24 hours, with LIVE-DEAD test kit (Invitrogen company), to cell dyeing, dead cell dyes for redness, and living cells dyes for green, detects cell survival rate, and result is as Figure 14.

As seen in Figure 14, matched group and blank carrier A u@SiO ₂group is after laser irradiation, and cell survival rate is without obvious decline; And the carrier A u@SiO of bag medicine carrying thing ₂-DOX after laser irradiation 12 hours and 24 hours, cell survival rate will decline gradually.This explanation carrier A u@SiO ₂-DOX is after laser irradiation, and the amycin that bag is carried discharges, and has played the effect of killing cancerous cell.Matched group is not to the cell of any drug treating, blank carrier A u@SiO ₂group is Au@SiO ₂the cell of processing, vehicle group is the Au@SiO of 5 micromoles per liter equipotent concentration ₂the cell that-DOX processes.

Embodiment 15

The present embodiment is used for illustrating cell membrane integrity evaluation after laser irradiation cell

1) cell culture

People's lung cancer A549 cell is inoculated in 35 millimeters of co-focusing imaging culture dishs, use DMEM fluid medium (streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls) in 37 ℃, in 5% CO2 gas incubator, cultivate phosphate buffered solution washed twice 24 hours.

2) cell membrane integrity detects

Two-photon Laser Scanning Confocal Microscope is for studying the integrity of lysosome membrane.

(1) add the Au@SiO containing 5 micromoles per liter equivalence doxorubicin concentration of preparation in embodiment 1 ₂the DMEM fluid medium (streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls) of-DOX is processed cell 12 hours;

(2) with containing the DMEM liquid culture of 5 mcg/ml acridine oranges (AO) based on 37 ℃ of incubated cells 15 minutes, phosphate buffered solution washing 3 times, then adds serum-free DMEM fluid medium;

(3) utilize the microscopical near-infrared laser of two-photon fluorescence (780 nano wave lengths, femtosecond pulse Maitai laser instrument, microscope is OlympusBX61W1, Japan) irradiate, laser power density is 24 and 48 watts/square centimeter, and irradiation time is 0,3,8 minute;

(4) under Laser Scanning Confocal Microscope, observe, 488 nanometers excite, and transmit respectively in 537 nanometers and the collection of 615 nanometers.Red membrane bubble structure is complete lysosome; If the danger signal of endochylema inner membrance balloon-shaped structure weakens, around lysosome, there is red green alternate or green membrane structure simultaneously, show that lysosome membrane structure sustains damage.In addition, observation of cell film edge emit bubble structure, the integrity degree of analysis of cells film, as Figure 15.

As seen in Figure 15, take in Au@SiO ₂the cancerous cell of-DOX is after high power (48 watts/square centimeter) laser irradiation, and lysosome membrane breaks, cell death; The following laser of low-power (24 watts/square centimeter) has no significant effect lysosome membrane, illustrates that carrier described in the present invention can be converted into heat by laser, kills cancerous cell by destroying lysosome membrane.

Embodiment 16

The present embodiment is for illustrating the impact of intracellular low power laser control drug release on tumor cell activity.The present embodiment selects dissimilar tumor cell, different lower powered laser, different irradiation time to investigate the chemotherapy effect of laserthermia and drug release induction.

1) cell culture

Select various human source tumor cell to comprise human lung carcinoma cell (A549), human breast cancer cell (MCF-7), human breast cancer cell (MDA-MB-231), human melanoma cell (A375), human cervical carcinoma cell (HeLa), human liver cancer cell (HepG2) and Human Prostate Cancer Cells (LNCap), with 3000 cells/well density, these cells are inoculated in respectively in 96 orifice plates, use DMEM fluid medium (to contain 10 % by weight hyclones, 1 % by weight penicillin, the streptomycin of 100 units per ml, the glutamine of 2 mM/ls) in 37 ℃, in 5% CO2 gas incubator, cultivate 24 hours, phosphate buffered solution washed twice.Add respectively the Au@SiO that contains 5 micromoles per liter equivalence amycin ₂prepared by-DOX(embodiment 1) and contain Au@SiO ₂(prepared by embodiment 1, the Au@SiO of its consumption and 5 micromoles per liter equivalence amycin ₂contained Au@SiO in-DOX ₂identical) complete medium (DMEM fluid medium, its streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls), and process respectively cell 12 hours.

2) laser irradiation

Abandon cell culture medium, add complete medium (DMEM fluid medium, its streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls).Select A549 cell, MCF-7 cell, MDA-MB-231 cell, A375 cell to irradiate, near infrared laser irradiates the cell (femtosecond laser 780 nanometers, power density: 24 watts/square centimeter) in orifice plate, and irradiation time is 3,5,8 minutes.Select HeLa cell, HepG2 cell, HNCap cell to irradiate, near infrared laser irradiates the cell (femtosecond laser 780 nanometers, power density: 12 watts/square centimeter) in orifice plate, and irradiation time is 5,10min.Respectively establish an experimental group and (be respectively the Au@SiO of above-mentioned 5 micromoles per liter equivalence amycin for taking in medicine group ₂groups of cells and Au@SiO that-DOX processes ₂the groups of cells of processing), take in medicine laser irradiation group, and have matched group (without drug treating with without laser irradiation group), separately establish an experimental group for adding 1 micromoles per liter amycin.

3) cell viability is measured

Irradiate the rear vigor that utilizes respectively CCK-8 test kit test cell for 24 hours.Every hole adds 110 microlitre mixed liquors, mixed liquor is containing 90 volume % complete medium (DMEM fluid mediums, its streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls) and the cell culture fluid of 10 volume %CCK-8, in incubator, hatch after 2 hours.In microplate reader, be determined at 450 nanometers absorbances, 600 nanometers light absorption values are reference wavelength; After every group of light absorption value deduction blank solution light absorption value, every hole respective value divided by matched group as cell viability.Every group arranges 6 parallel holes, and every group of experiment in triplicate.

Evaluate after laser irradiation Au@SiO ₂with Au@SiO ₂the impact of-DOX on cell viability; Whether analysis of cells vigor changes is that drug release causes.Can find out Au@SiO by Figure 16 (A, B) ₂group is after low-power (24 watts/square centimeter and 12 watts/square centimeter) laser irradiation, and cell viability is all without obviously declining, and shows that shortage of heat that carrier changes into the laser of two kinds of power is to kill cancerous cell; And Au@SiO ₂-DOX group is after two kinds of power lasers irradiate, and cell viability all declines gradually in follow-up 24 hours, illustrates that the amycin that wherein bag carries discharges gradually after laser irradiation, has caused the decline of cell viability, reaches the effect of chemotherapy.

Embodiment 17

The present embodiment is the impact on tumor cell activity for the chemotherapy double-mode that high power laser light is described realizes drug release in thermotherapy and cell simultaneously.

1) cell culture

Select three-type-person source tumor cell to comprise the cells such as human lung carcinoma cell (A549), human breast cancer cell (MCF-7), human melanoma cell (A375), with 3000 cells/well density, these cells are inoculated in respectively in 96 orifice plates, use complete medium (DMEM fluid medium, its streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls) in 37 ℃, in 5% CO2 gas incubator, cultivate phosphate buffered solution washed twice 24 hours.Add respectively the Au@SiO that contains 5 micromoles per liter equivalence amycin ₂prepared by-DOX(embodiment 1) and contain Au@SiO ₂(prepared by embodiment 1, the Au@SiO of its consumption and 5 micromoles per liter equivalence amycin ₂contained Au@SiO in-DOX ₂identical) complete medium (DMEM fluid medium, its streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls), and process respectively cell 12 hours.

2) laser irradiation

Abandon cell culture medium, add complete medium (DMEM fluid medium, its streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls), select A549, MCF-7, A375 cell, use near infrared laser to irradiate cell (femtosecond laser 780 nanometers in orifice plate, power density: 48 watts/square centimeter), irradiation time is 12 minutes.Respectively establish an experimental group and (be respectively the Au@SiO of above-mentioned 5 micromoles per liter equivalence amycin for taking in medicine group ₂groups of cells and Au@SiO that-DOX processes ₂the groups of cells of processing), take in medicine laser irradiation group, and have matched group (without drug treating with without laser irradiation group).

3) cell viability is measured

Irradiate the rear vigor that utilizes respectively CCK-8 test kit test cell for 12 hours.Every hole adds 110 microlitre mixed liquors, mixed liquor is containing 90 volume % complete medium (DMEM fluid mediums, its streptomycin that contains 10 % by weight hyclones, 1 % by weight penicillin, 100 units per ml, the glutamine of 2 mM/ls) and the cell culture fluid of 10 volume %CCK-8, in incubator, hatch after 2 hours.In microplate reader, be determined at 450 nanometers absorbances, 600 nanometers light absorption values are reference wavelength; After the light absorption value of every group of light absorption value deduction blank solution (only adding 90 volume % complete mediums and 10 volume %CCK, acellular), every hole respective value divided by matched group (without drug treating, without the groups of cells of laser irradiation) as cell viability.Every group arranges 6 parallel holes, and every group of experiment in triplicate.

Evaluate after high power laser light irradiation Au@SiO ₂with Au@SiO ₂the impact of-DOX on cell viability; Whether analysis of cells vigor changes is that the synergism that thermotherapy, hot ancillary drug discharge causes.Can find out Au@SiO by Figure 17 ₂group is after 48 watts of/square centimeter of high power laser lights irradiate, and cell viability obviously declines, and shows that the heat that carrier changes into this power laser can kill most cancerous cell; After laser irradiation, compare carrier, Au@SiO simultaneously ₂-DOX group cell viability significantly declined in follow-up 12 hours, illustrated that it kills the cooperative effect that tumor cell mode is photo-thermal and drug release chemotherapy, and carrier can be changed by photo-thermal on the one hand, kills most of tumor cell, plays a leading role; The amycin that the opposing party's bread carries discharges rapidly after laser irradiation, has also reduced cell viability.

Embodiment 18

The present embodiment is used for illustrating Au@SiO ₂-DOX suppresses the effect of tumor growth on animal model for tumour.

1) animal inoculation

Get

6,7 week age, body weight is the male Balb C mice of 17-19 gram.By the number of cells of exponential phase (about 10 ⁶individual) to be inoculated in mice upper limb armpit subcutaneous for mankind mastopathy cell (MCF-7), observes and record tumor growth situation every day.After inoculated tumour 10-14 days, treat that tumor mass grows to diameter comparatively suitable at 2 millimeters ~ 20 millimeters.

2) laser irradiation

Give the Au@SiO of preparation in embodiment 1 to the tumor locus of tumor-bearing mice by tail vein injection ₂-DOX(dosage is equivalent to 50 mg/kg body weight), then use wavelength 780 nanometers, power 600 milliwatts, the laser scanning uniform irradiation tumor locus that hot spot is 2 millimeters, position, every place is irradiated 15 minutes.

2) after laser irradiation, tumor growth suppresses situation

After laser irradiation, taking in medicine 24 hours, 72 hours, 120 hours respectively, measuring tumor average diameter and be respectively 8 millimeters, 5 millimeters and 2 millimeters.And the tumor average diameter of medication mice matched group (inoculated tumour gives normal saline injection) is not respectively 10 millimeters, 12 millimeters and 15 millimeters.Then, dispose standard by local animal and put to death mice, take out tumor tissues, weigh, confirm neoplasm necrosis by observation.Partial tumors is organized in 10% formalin and fixes 24 hours, by paraffin embedding, section (5 micron thickness), under inverted microscope, observe tumor tissue section in Au@SiO ₂the gathering situation of-DOX, as Figure 18.

Carrier A u@SiO of the present invention as seen in Figure 18 ₂-DOX can be accumulated in tumor tissues after entering in tumor-bearing mice body; And after laser irradiation, can cause the tumor tissue necrosis of carrier periphery.

More than describe the preferred embodiment of the present invention in detail; but the present invention is not limited to the detail in above-mentioned embodiment, within the scope of technical conceive of the present invention; can carry out multiple simple variant to technical scheme of the present invention, these simple variant all belong to protection scope of the present invention.

It should be noted that in addition, each concrete technical characterictic described in the above-mentioned specific embodiment, in reconcilable situation, can combine by any suitable mode, for fear of unnecessary repetition, the present invention is to the explanation no longer separately of various possible compound modes.

In addition, also can carry out combination in any between various embodiment of the present invention, as long as it is without prejudice to thought of the present invention, it should be considered as content disclosed in this invention equally.

Claims

1. a pharmaceutical carrier, is characterized in that, this pharmaceutical carrier comprises gold nanorods, and is coated on the meso-porous titanium dioxide silicon layer on gold nanorods surface.

2. pharmaceutical carrier according to claim 1, wherein, the length of described gold nanorods is 5-200 nanometer; The draw ratio of described gold nanorods is 1.5-20.

3. pharmaceutical carrier according to claim 1 and 2, wherein, the thickness of described meso-porous titanium dioxide silicon layer is 3-50 nanometer.

4. pharmaceutical carrier according to claim 3, wherein, the aperture of described meso-porous titanium dioxide silicon layer is 1-20 nanometer, the specific surface area of described pharmaceutical carrier is 1-150 meters squared per gram, pore volume be 0.05-10 cubic centimetre/gram.

5. the preparation method of a pharmaceutical carrier, it is characterized in that, the method comprises the following steps: in the gold nanorods aqueous solution that contains cetyl trimethyl ammonium bromide, under the condition that is 8.5-11 at pH value, add the alcoholic solution of esters of silicon acis to react, and carry out solid-liquid separation, obtain surface and be coated with the gold nanorods of meso-porous titanium dioxide silicon layer.

6. method according to claim 5, wherein, in the aqueous solution of described gold nanorods, take the gold nanorods concentration of gold element as 10 micromoles per liter ~ 2.5 mM/l, the length of described gold nanorods is 5-200 nanometer; The draw ratio of described gold nanorods is 1.5-20.

7. according to the method described in claim 5 or 6, wherein, respectively in gold element and element silicon, the gold nanorods in the aqueous solution of described gold nanorods and the mol ratio of described esters of silicon acis are 1:1-50; Take the described gold nanorods of gold element and the mol ratio of cetyl trimethyl ammonium bromide as 1:1-20.

8. method according to claim 5, wherein, the condition of described reaction also comprises: the temperature of contact is 4-95 ℃, the time of contact is 0.04-10 days.

9. method according to claim 5, wherein, described esters of silicon acis is quanmethyl silicate, tetraethyl orthosilicate, one or more in silicic acid orthocarbonate and silicic acid four butyl esters.

10. method according to claim 5, wherein, described alcohol is that methanol, ethanol or other molecular weight are less than 200, be one or more of the alcohol apoplexy due to endogenous wind of liquid under normal temperature and pressure.

11. 1 kinds of pharmaceutical compositions, described pharmaceutical composition comprises carrier and loads on the medicine on carrier, it is characterized in that, described carrier is the pharmaceutical carrier described in any one in claim 1-4, or is by the prepared pharmaceutical carrier of method described in any one in claim 5-10.

12. pharmaceutical compositions according to claim 11, wherein, described medicine is one or more in amycin, paclitaxel, vinblastine, cisplatin, camptothecine, DNA and siRNA.

13. according to the pharmaceutical composition described in claim 11 or 12, and wherein, take the weight of described carrier as benchmark, the carrying drug ratio of described pharmaceutical composition is 1-40 % by weight.

The application of pharmaceutical composition in pharmaceutical carrier in 14. claim 1-4 described in any one and/or claim 11-13 described in any one in the medicine for the preparation of diagnosis and/or treatment tumor.