CN109806394A - A kind of mesoporous silicon oxide drug delivery system and its application - Google Patents
A kind of mesoporous silicon oxide drug delivery system and its application Download PDFInfo
- Publication number
- CN109806394A CN109806394A CN201711151069.4A CN201711151069A CN109806394A CN 109806394 A CN109806394 A CN 109806394A CN 201711151069 A CN201711151069 A CN 201711151069A CN 109806394 A CN109806394 A CN 109806394A
- Authority
- CN
- China
- Prior art keywords
- msn
- mesoporous silicon
- tpgs
- drug
- preparation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Abstract
The present invention relates to a kind of mesoporous silicon oxide drug delivery system and its application, the research for the stimuli responsive type drug delivery system that the new chemotherapeutic and photo-thermal therapy for belonging to medical treatment field combine.The present invention is that the MSN for having two-dimentional tubulose cellular structure is prepared in template, and removes CTAB template using acid extraction method using adriamycin as model drug, using CTAB, will remove the MSN of template as pharmaceutical carrier.By disulfide bond by Cypate grafting and modifying to carrier surface, the efficient photothermal conversion efficiency of medicine-carried system, and physiological stability and biocompatibility in outermost layer cladding hydrophilic material TPGS to improve medicine-carried system are assigned.On the basis of restoring response type drug release by chemotherapy in conjunction with photo-thermal therapy, increase the sensibility of intake and intracellular delivery system reduction responsiveness drug release of the cell to targeting vector, accelerate the release of drug, to realize the synergy of chemotherapy and photo-thermal therapy, the toxic side effect of chemotherapeutics is further decreased.
Description
Technical field
The invention belongs to pharmaceutical technology field, it is related to a kind of mesoporous silicon oxide drug delivery system and its application.Specifically
It is related to the building and its application of the mesoporous silicon oxide medicine-carried system of the disulfide bond connection photo-thermal dyestuff Cypate coated by TPGS.
The mesoporous silicon oxide drug delivery system can be used as the stimuli responsive type drug that new chemotherapeutic and photo-thermal therapy combine
Delivery system.
Background technique
Drug delivery system reaches therapentic part and usually requires the regular hour from being administered to, classical drug delivery system by
The ability of initiation drug release, can generate drug after lacking the specific recognition to therapentic part and reaching therapentic part
It discharges in advance, and then leads to the low and all kinds of toxic side effect of bioavilability.
For the drug concentration for effectively improving lesion tissue, dosage is reduced, weakens anticancer drug normal tissue organ
Toxic side effect, the stimuli responsive type drug delivery system for being caused drug release using the special physiological feature of target area become
One of the hot spot of galenic pharmacy research.It has designed in recent years and has had the properties such as sensitive reduction, pH sensitivity, enzyme sensitivity and photaesthesia
Stimuli responsive type drug delivery system, wherein reduction sensitivity apply relatively broad at present.Glutathione (GSH) is used as cell
The GSH concentration of interior important reducing substances, usual extracellular fluid and blood plasma only has 2-20 μM, and the GSH concentration of intracellular fluid
Can reach 1-10mM, can achieve if the GSH concentration in some special lesion tissues (such as tumor tissues) 3 times of normal tissue with
On.Since disulfide bond is easily broken under the GSH effect of high concentration, reduction responsive type drug delivery system can be designed accordingly.
Although stimuli responsive type drug delivery system can efficiently control the release in advance of drug, make so that the poison for reducing drug is secondary
With, but still come with some shortcomings, specifically include that the drugloading rate of (1) drug is lower, the drug big for dosage cannot expire
The needs of sufficient clinical medicine dose；(2) even if galenic pharmacy means is taken to improve drugloading rate, medicine-carried system is in storage or transportational process
Middle physical stability decline, cannot effectively inhibit drug to recrystallize or reassociate, cause the solubility of drug in physiological conditions
Or rate of release is substantially change, reduction significant in efficacy of drug etc.；(3) drugloading rate is excessive or the increase of dosage, all
It will lead to the generation of the toxic reaction and multidrug resistance of drug.Therefore, the suitable clinical medicine dose that can satisfy is found to want
The drugloading rate asked, and drug-loading system stabilization and drug delivery system significant in efficacy are the hot spot and hardly possible in Pharmaceutical study field
Point.
The appearance of porous material opens new road for the design of stimuli responsive type drug delivery system.According to the world
Purely with the definition of applied chemistry federation (IUPAC), porous material is divided into three classes according to the size in aperture: aperture is less than 2nm
Be known as poromerics, aperture between 2nm-50nm be known as mesoporous material, aperture greater than 50nm then be known as macropore material
Material.In numerous nano inorganic porous carriers, aperture that mesoporous silicon oxide is of moderate size by it, huge specific surface area
With high porosity, the adjustable mesoporous pore size of uniform and ordered and pattern, good biocompatibility and surface easy functional modification
Many advantages, such as hydrophily silanol group, can be used as ideal nano-medicament carrier, be widely used in new drug delivery systems
Research in.It finds that mesoporous silicon oxide can significantly improve the dissolution rate of insoluble drug in research, and utilizes its hole
Wall improves the physical stability of drug to the restriction effect of drug molecule.
The problems such as treatment of lesion tissue is still limited by drug merely, and is easy to cause drug resistance, because
This photo-thermal therapy (thermotherapy) becomes a kind of new methods of adjuvant treatment.Its cardinal principle mainly turns using with high light heat
The material or carrier for changing efficiency, are injected into inside of human body, are gathered near tumor tissues using targeting identification technology,
And under the irradiation of external light source (usually near infrared light), optothermal material efficiently converts luminous energy to thermal energy and then generates and controls
Treatment effect.The finer and close lesion tissue of some structures (such as tumor tissues) heat dissipation is slow and more sensitive to fuel factor, so light
Heat therapy can be such that the local temperature of lesion tissue increases, and the influence to normal cell is relatively small.Near infrared light and it is visible and
Ultraviolet light, which is compared, has stronger body tissue penetrability, provides possibility for the photo-thermal therapy for tumor of Noninvasive.Light
The advantages of heat therapy, includes, first is that reducing the pain that patient is subjected to；Second is that shortening treatment time (a few minutes) and therapeutic effect
Obviously.The core of photo-thermal therapy is the nano material of superpower photothermal conversion efficiency.Photosensitizer currently used for photo-thermal therapy is main
For inorganic nanoparticles such as gold nanorods, carbon nanotube and graphene etc. and organic fluorescent dye, such as indocyanine green
(ICG), porphyrin, phthalocyanine etc..Wherein ICG is a kind of functional dye molecule, is U.S. FDA approval with near-infrared characteristic
The optical diagnostics factor, while be also a kind of photo-thermal therapy light absorber.But since its concentration dependent is assembled, molecule is steady
It is qualitative poor, and its application is limited with properties such as the non-specific bindings of albumen.Cypate (Figure 1A in Detailed description of the invention)
It is the carboxy derivatives of ICG (Figure 1B), fluorescence can be generated near infrared region, absorption maximum and launch wavelength are respectively
780nm and 810nm.Relative to ICG, the stability of Cypate is significantly improved, and surface has modifiable carboxylic group.
Medicine is not carried with the mesoporous silicon oxide of the disulfide bond connection photo-thermal dyestuff Cypate of TPGS cladding in the prior art
The stimuli responsive type drug delivery system that system is combined for chemotherapy and photo-thermal therapy.
Summary of the invention
The object of the present invention is to provide a kind of reduction response types for having near infrared light hot-cast socket ability, to realize that drug exists
The stimuli responsive type release of target tissue and chemotherapy, the synergistic effect of photo-thermal therapy enhance therapeutic effect and reduce toxic side effect.
The present invention by near-infrared organic carbon cyanine type dye Cypate (3- (2-carboxyethyl) -2- ((1E, 3E, 5E,
7E)-7-(3-(2-carboxyethyl)-1,1-dimethyl-1H-benzo[e]indol-2(3H)-ylidene)hepta-
1,3,5-trien-1-yl) -1,1-dimethyl-1H-benzo [e] indol-3-ium) it is modified by disulfide bond mesoporous two
The channel surfaces of silica support, constructing, there is the reduction response type of near infrared light hot-cast socket ability to gate system, and in body
The surface of system carries out TPGS cladding, improves the dispersion stabilization of whole system.
Drug delivery system of the present invention is the mesoporous silicon oxide that TPGS is coated on to disulfide bond grafting Cypate
Nanoparticle and be made, which has reduction response type drug release ability and the lower change thermotherapy synergy of near-infrared irradiation.
The present invention coats the mesoporous silicon dioxide nano particle (MSN-SS-Cy/ that disulfide bond is grafted Cypate by TPGS
TPGS) Cypate covalence graft is coated TPGS in meso-porous titanium dioxide silicon face, and using film hydration method using disulfide bond
In outermost layer, the Nano medication of the thermotherapy obtained having under near infrared light and reduction response type drug release ability delivers system
System.
The technical solution adopted in the present invention is as follows:
1) preparation of mesoporous silicon dioxide nano particle (MSN):
It is template using cetyl trimethylammonium bromide (CTAB), the MSN for having two-dimentional tubulose cellular structure is made.
2) preparation of sulfhydrylation mesoporous silicon dioxide nano particle (MSN-SH):
By MSN support dispersion into dehydrated alcohol, under nitrogen protection with (3- mercaptopropyi) trimethoxy silane
(MPTMS) the mesoporous silicon carrier MSN-SH of sulfhydrylation modification is made in back flow reaction, then removes CTAB mould using acid extraction method
Plate will expose the MSN of mesopore orbit as pharmaceutical carrier.
3)MSN-SS-NH2Preparation:
MSN-SH carrier and S- (2- aminoethanethio) -2- thiopyridine (Py-SS-NH2) back flow reaction, in mesoporous silicon table
Face introduces disulfide bond, and the MSN carrier (MSN-SS-NH that amino terminal is connected by disulfide bond is made2)。
4) preparation of MSN-SS-Cy:
Cypate is dissolved in dimethyl sulfoxide (DMSO), 1- ethyl-(3- dimethylaminopropyl) carbon two is being added
After inferior amine salt hydrochlorate (EDC) and n-hydroxysuccinimide (NHS) activate 1 hour, with MSN-SS-NH2Carrier reflux is prepared into
To the MSN carrier (MSN-SS-Cy) for being grafted Cypate by disulfide bond.
5) preparation of MSN-SS-Cy/TPGS
Upper TPGS is coated using the film hydration method of improvement, the MSN-SS-Cy (MSN-SS-Cy/ of surface cladding TPGS is made
TPGS)。
In step (1): micella is formed using CTAB and is used as pore-foaming agent, ethyl orthosilicate (TEOS) is as silicon source, and 80 DEG C to 100
DEG C reaction, TEOS hydrolysis generate silica.
In step (2): the volume ratio of sulfhydrylization reagent MPTMS and first step addition TEOS are between 1: 7 to 1: 3.
In step (3): reaction reacts 24-48h under the conditions of room temperature (25 DEG C ± 2 DEG C).
In step (4): using EDC and NHS as condensing agent.
In step (5): TPGS density is (1.00-1.10) * 103kg/m3。
Specifically, in the preparation of MSN-SS-Cy/TPGS of the present invention:
1) preparation of the MSN of two-dimentional tubulose cellular structure:
The present invention forms micella using CTAB and is used as pore-foaming agent, and ethyl orthosilicate (TEOS) is as silicon source, and 80 DEG C to 100 DEG C
Lower hydrolysis TEOS is made, and it is described that specific preparation process is as follows:
0.3g -0.7g CTAB is added in 200mL -300mL distilled water, the magnetic agitation in 80-100 DEG C of water-bath
(700rpm) 30min or more is kept stirring condition after it is completely dissolved, and the sodium hydroxide that the 2mol/L of 1mL -2mL is added is molten
Liquid continues stirring and keeps after reacting 15-30min, the TEOS of 2mL -3mL is slowly added dropwise, keeps its evenly dispersed in the reaction system,
Reaction stops after keeping 2-3h, is cooled to room temperature to reaction solution, sample is collected by centrifugation, and is washed using distilled water and dehydrated alcohol,
It is spare to collect product.
2) preparation of sulfhydrylation mesoporous silicon dioxide nano particle (MSN-SH)
MSN carrier is evenly spread in 50mL -60mL dehydrated alcohol, the MPTMS of 0.3mL -0.8mL is then added,
Flow back 8-16h under 80 DEG C of nitrogen protections, to after reaction, sample be collected by centrifugation (8000rpm, 10min), with anhydrous second
Alcohol washs three times, removes surfactant templates using acid extraction method later, sample is added to 18mL concentrated hydrochloric acid (mass fraction
37%) and in the mixed solution of 320mL methanol, flow back 12h in 80 DEG C of water-baths, be centrifuged after extraction (8000rpm,
The sample for 10min) collecting removing template, is washed 3 times with dehydrated alcohol, spare.
3)MSN-SS-NH2Preparation
By the MSN-SH support dispersion of 500mg -600mg removing template into the dehydrated alcohol of 20mL -30mL, then it is added
The Py-SS-NH of 200mg -300mg2, mixture is stirred to react 24-48h in 500rpm under the conditions of room temperature (25 DEG C ± 2 DEG C),
Product collects precipitating by centrifugation (8000rpm, 10min), and dehydrated alcohol is cleaned spare.
4) preparation of MSN-SS-Cy:
The Cypate of 100mg -200mg is dissolved in 100mL -200mL DMSO, be added 50mg -100mg EDC and
After 100mg -200mg NHS is activated 1 hour, the MSN-SS-NH of 500mg -600mg is continuously added2Carrier.It is anti-in 500rpm stirring
It answers 24 hours, product collects precipitating by centrifugation (8000rpm, 10min), and dehydrated alcohol is cleaned spare.
5) cladding of TPGS
The film hydration method preparation that the MSN-SS-Cy (MSN-SS-Cy/TPGS) of TPGS cladding passes through improvement.Weigh 40mg-
50mg TPGS is dissolved in 100mL -150mL water.Separately 80mg -130mg hydrophobic MSN-SS-Cy carrier is mixed with 5mL chloroform
It closes, ultrasonic 2min disperses it well.TPGS solution and suspension are mixed.Be evaporated under reduced pressure remove chloroform after, TPGS with
MSN-SS-Cy forms uniform film.7.4 phosphate buffer of pH and ultrasound 2min that 100mL is added disperse again, and centrifugation removes
Excessive TPGS is removed, sediment is MSN-SS-Cy/TPGS.
MSN-SS-Cy/TPGS nano-carrier prepared by the present invention, particle diameter distribution 250-350nm, Zeta potential are -25
To -15mv.(being measured under pH=7.4 condition phosphate buffered saline solution).
The present invention assigns nano-carrier using the mesoporous silicon dioxide nano particle of the disulfide bond grafting Cypate of TPGS cladding
In the stimuli responsive type drug release ability and chemotherapy of target tissue, the synergy of photo-thermal therapy and reduce toxic side effect.It is described
Drug can be with are as follows: adriamycin, camptothecine, taxol etc..
Wherein, the proportionate relationship between drug and the mesoporous silicon dioxide nano particle of the disulfide bond grafting Cypate of TPGS cladding
Are as follows: between mass ratio 1:1 to 1:5.
By taking adriamycin as an example:
Adriamycin can be loaded into the MSN-SH of step 2) preparation, then according still further to the MSN- of subsequent step preparation drug containing
SS-Cy/TPGS nano-carrier.
The ratio of the mesoporous silicon dioxide nano particle of adriamycin and the disulfide bond grafting Cypate of TPGS cladding are as follows: mass ratio
Between 1:1 to 1:5.
Specifically adriamycin is loaded into the preparation of MSN-SH:
20mg -30mg DOX is dissolved in 4mL -7mL distilled water, under room temperature stir about 1h, 80mg-is then added
120mg MSN-SH is ultrasonically treated 1h, and the PBS solution of 8mL -12mL pH7.4 is then added, and continues stirring for 24 hours.It will carry DOX's
MSN-SH is centrifuged 5min precipitated nanocrystals particle in 6000, and the DOX for removing MSN-SH adsorption is washed with the PBS of pH 5.0, is subtracted
It press dry dry 10-20h and removes organic solvent to get drug-carrying nanometer particle.
The present invention utilizes the inner lumen structure of carrier and outer using mesoporous silicon oxide as the carrier of insoluble drug
The restriction effect of layer mesoporous wall realizes the efficient loading and stable dispersion of drug.Then photo-thermal dyestuff Cypate is passed through two
Sulfide linkage grafts on the mesopore orbit surface of mesoporous silicon oxide, becomes " reduction/photothermal response door-controlled type " switch, assigns the medicine
Object delivery system restores the double characteristic of response type drug release and photo-thermal synergy under near infrared light, when delivery system is targeted to lesion group
After knitting, lesions position is irradiated near infrared light, luminous energy can be efficiently converted into heat by photo-thermal dyestuff Cypate, make office
Portion's temperature increases, and cell increases the intake of targeting vector, while intracellular delivery system reduction responsiveness drug release is quick
Perception increases, and accelerates the release of drug, to realize the synergy of chemotherapy and photo-thermal therapy, further decreases chemotherapeutics
Toxic side effect.
Detailed description of the invention
Fig. 1 is the molecular formula of Cypate (A) and ICG (B)；
Fig. 2 is MSN transmission electron microscope results prepared by embodiment 1；
The MSN-SS-NH of Fig. 3 is the MSN of the preparation of embodiment 1, prepared by embodiment 4 MSN-SH, the preparation of embodiment 62And
The N2 adsorption adsorption isotherm of MSN-SS-Cy/TPGS prepared by MSN-SS-Cy and embodiment 10 prepared by embodiment 8；Fig. 4 is
The MSN-SS-NH of MSN-SH, the preparation of embodiment 6 prepared by the MSN of the preparation of embodiment 1, embodiment 42And the preparation of embodiment 8
The pore-size distribution of MSN-SS-Cy/TPGS prepared by MSN-SS-Cy and embodiment 10；
The MSN-SS-NH that Fig. 5 is MSN-SH prepared by embodiment 4 and prepared by embodiment 62, the MSN- of the preparation of embodiment 10
The Zeta potential of SS-Cy/TPGS；
Fig. 6 is MSN-SH prepared by embodiment 4, MSN-SS-NH prepared by embodiment 62The MSN- prepared with embodiment 8
The TGA data of MSN-SS-Cy/TPGS prepared by SS-Cy and embodiment 10；
The near-infrared of Fig. 7 is illumination power the MSN-SS-Cy/TPGS that prepared by embodiment 10 when being 2W heats up data；
Fig. 8 is heating of the various concentration MSN-SS-Cy/TPGS of the preparation of embodiment 10 in the case where different near infrared lights are according to power
Data；
Fig. 9 is pH and the reduction response type release for the MSN-SS-Cy/TPGS drug delivery system that DOX is loaded；
Figure 10 is drug release percentage of the MSN-SS-Cy/TPGS of DOX loading under different release conditions；
Figure 11 is DOX solution group, and MSN-SS-Cy/TPGS empty vectors, MSN-SS-Cy/TPGS medicine-carried system is in 808nm
NIR irradiation under 4T1 breast cancer cell survival rate.
Specific embodiment
Embodiment 1
It accurately weighs 0.5g CTAB to be added in the beaker of 500mL, 240mL distilled water is added into beaker, in 80 DEG C of water
Magnetic agitation (700rpm) is kept stirring condition, Xiang Shangshu mixed solution continuously adds 1.75mL's to pellucidity in bath
The sodium hydroxide solution of 2mol/L continues that the TEOS of 2.5mL is added dropwise after stirring 15min, continues to stir 2h under 80 DEG C of water-baths.
It is cooled to room temperature to reaction solution, centrifugation (7500rpm, 7min) collection obtains white solid, is filled using distilled water and dehydrated alcohol
Divide washing, clean sample is added in the mixed solution of 18mL concentrated hydrochloric acid (mass fraction 37%) and 320mL methanol, 80
Flow back 12h in DEG C water-bath, and the sample that (8000rpm, 10min) collects removing template is centrifuged after extraction, is washed with dehydrated alcohol
It washs 3 times, crosses 80 meshes after dry, obtaining white product is mesoporous silicon dioxide nano carrier (MSN).
Mesoporous silicon dioxide nano carrier (MSN) obtained is in spheroid shape, and uniform particle diameter has apparent arranged in parallel
Mesopore orbit, duct size are conducive to efficient loading and " gate " release of drug in 2nm to 4nm.
Embodiment 2
It accurately weighs 0.6g CTAB to be added in the beaker of 500mL, 250mL distilled water is added into beaker, in 80 DEG C of water
Magnetic agitation (700rpm) is kept stirring condition, Xiang Shangshu mixed solution continuously adds 1.5mL's to pellucidity in bath
The sodium hydroxide solution of 2mol/L continues that the TEOS of 2.5mL is added dropwise after stirring 15min, continues to stir 2h under 80 DEG C of water-baths.
It is cooled to room temperature to reaction solution, centrifugation (7500rpm, 7min) collection obtains white solid, is filled using distilled water and dehydrated alcohol
Divide washing, clean sample is added in the mixed solution of 18mL concentrated hydrochloric acid (mass fraction 37%) and 320mL methanol, 80
Flow back 12h in DEG C water-bath, and the sample that (8000rpm, 10min) collects removing template is centrifuged after extraction, is washed with dehydrated alcohol
It washs 3 times, is sieved with 100 mesh sieve after dry, obtaining white product is mesoporous silicon dioxide nano carrier (MSN).
Embodiment 3
It accurately weighs 0.7g CTAB to be added in the beaker of 500mL, 260mL distilled water is added into beaker, in 80 DEG C of water
Magnetic agitation (700rpm) is kept stirring condition, Xiang Shangshu mixed solution continuously adds the 2mol/L of 2mL to pellucidity in bath
Sodium hydroxide solution, continue that the TEOS of 2.75mL is added dropwise after stirring 15min, continue to stir 2h under 80 DEG C of water-baths.To anti-
Liquid is answered to be cooled to room temperature, centrifugation (7500rpm, 7min) collection obtains white solid, sufficiently washes using distilled water and dehydrated alcohol
It washs, clean sample is added in the mixed solution of 18mL concentrated hydrochloric acid (mass fraction 37%) and 320mL methanol, in 80 DEG C of water
Flow back 12h in bath, and the sample that (8000rpm, 10min) collects removing template is centrifuged after extraction, washs 3 with dehydrated alcohol
It is secondary, 120 meshes are crossed after dry, obtaining white product is mesoporous silicon dioxide nano carrier (MSN).
Embodiment 4
It weighs MSN carrier 75mg obtained in embodiment 1 to evenly spread in 50mL dehydrated alcohol, 1.0mL is then added
MPTMS, flow back 13h under 80 DEG C of nitrogen protections, to after reaction, sample is collected by centrifugation (8000rpm, 10min),
It is washed three times with dehydrated alcohol, then sample is added to 18mL concentrated hydrochloric acid (mass fraction 37%) and the mixing of 320mL methanol is molten
In liquid, flow back 12h in 80 DEG C of water-baths, the sample that (8000rpm, 10min) collects removing template is centrifuged after extraction, with nothing
Water-ethanol washs 3 times, and gained sample is sulfhydrylation mesoporous silicon dioxide nano particle (MSN-SH).
Embodiment 5
It weighs MSN carrier 70mg obtained in embodiment 1 to evenly spread in 55mL dehydrated alcohol, 1.2mL is then added
MPTMS, flow back 13h under 80 DEG C of nitrogen protections, to after reaction, sample is collected by centrifugation (8000rpm, 10min),
It is washed three times with dehydrated alcohol, then sample is added to 18mL concentrated hydrochloric acid (mass fraction 37%) and the mixing of 320mL methanol is molten
In liquid, flow back 12h in 80 DEG C of water-baths, the sample that (8000rpm, 10min) collects removing template is centrifuged after extraction, with nothing
Water-ethanol washs 3 times, and gained sample is sulfhydrylation mesoporous silicon dioxide nano particle (MSN-SH).
Embodiment 6
MSN-SH carrier obtained in 500mg embodiment 4 is weighed, is distributed in the dehydrated alcohol of 25mL, is added later
The Py-SS-NH of 200mg2, mixture is stirred to react for 24 hours under the conditions of room temperature (25 DEG C ± 2 DEG C) in 500rpm, product pass through from
The heart (8000rpm, 10min) collects precipitating, and MSN-SS-NH is cleaned and obtained to dehydrated alcohol2。
Embodiment 7
MSN-SH carrier obtained in 600mg embodiment 4 is weighed, is distributed in the dehydrated alcohol of 30mL, is added later
The Py-SS-NH of 300mg2, mixture is stirred to react for 24 hours under the conditions of room temperature (25 DEG C ± 2 DEG C) in 500rpm, product pass through from
The heart (8000rpm, 10min) collects precipitating, and MSN-SS-NH is cleaned and obtained to dehydrated alcohol2。
Embodiment 8
100mg Cypate is weighed to be dissolved in 100mL DMSO, after 60mg EDC and 100mg NHS activation 1h is added,
Continuously add MSN-SS-NH obtained in 500mg embodiment 72Carrier.It is stirred to react for 24 hours in 500rpm, product is by centrifugation
(8000rpm, 10min) collects precipitating, and MSN-SS-Cy carrier is cleaned and obtained to dehydrated alcohol.
Embodiment 9
It weighs 120mg Cypate to be dissolved in 100mL DMSO, being added, 80mg EDC and 120mg NHS activation one are small
Shi Hou continuously adds MSN-SS-NH obtained in 600mg embodiment 72Carrier.It is stirred to react for 24 hours in 500rpm, product passes through
It is centrifuged (8000rpm, 10min) and collects precipitating, MSN-SS-Cy carrier is cleaned and obtained to dehydrated alcohol.
Embodiment 10
40mg TPGS is first weighed to be dissolved in 100mL water.The MSN-SS-Cy prepared in 80mg embodiment 8 is separately weighed to carry
Body is placed in the ultrasonic disperse 2min in 5mL chloroform.TPGS solution and suspension are mixed.It is evaporated under reduced pressure after removing chloroform, TPGS
Uniform film is formed with MSN-SS-Cy.7.4 phosphate buffer of pH and ultrasound 2min that 100mL is added disperse again, are centrifuged
Excessive TPGS is removed, sediment is MSN-SS-Cy/TPGS.
The heating of the near-infrared more than 40 DEG C may be implemented in system, and heating ability is good, is realizing the photo-thermal to tumor tissues
While treatment, promote the release of anticarcinogen.
Embodiment 11
50mg TPGS is first weighed to be dissolved in 120mL water.The MSN-SS-Cy prepared in 100mg embodiment 9 is separately weighed to carry
Body is placed in the ultrasonic disperse 2min in 10mL chloroform.TPGS solution and suspension are mixed.It is evaporated under reduced pressure after removing chloroform,
TPGS and MSN-SS-Cy form uniform film.7.4 phosphate buffer of pH and ultrasound 2min that 100mL is added disperse again,
It is centrifuged off excessive TPGS, sediment is MSN-SS-Cy/TPGS.
Embodiment 12
The MSN-SS-Cy/TPGS for taking 5mg DOX to load carries out extracorporeal releasing experiment: pH in different drug release media
5.0PBS, pH 7.4PBS solution, pH 5.0PBS are added 10mM GSH and pH 7.4PBS and 10Mm GSH are added.Dissolution medium
Volume is 1mL, and temperature is 37.5 DEG C, revolving speed 135rpm, is to measure absorbance at 480nm in wavelength, calculates under different time
Extracorporeal releasing quantity of adriamycin under the conditions of different pH medium and various concentration GSH.
The result shows that the MSN-SS-Cy/TPGS that DOX is loaded is in pH 5.0PBS solution (simulation tumour weak acid environment)
Rate of release ratio pH 7.4PBS solution (simulation normal tissue environment) in it is fast.Compared with GSH group is not added, medicine after GSH is added
The rate of release of object obviously increases, and illustrates that the medicine-carried system of preparation has apparent reduction response type drug release feature, related data is shown in
Fig. 9.
Embodiment 13
The MSN-SS-Cy/TPGS for taking 1mg DOX to load is distributed in the pH 5.0PBS of 1mL, the concentration of GSH in GSH group
For 10mM, temperature is 37.5 DEG C, revolving speed 135rpm.For NIR group, in 0h and 2h, sample is taken out, with the NIR of 808nm
Light is irradiated 2min, power 2W.After 2h and 4h, wavelength be 480nm place measurement absorbance, calculating different condition under Ah
The extracorporeal releasing quantity of mycin.
The result shows that DOX load MSN-SS-Cy/TPGS GSH exist or NIR light irradiation condition under, the release of drug
Rate obviously increases compared with the control group, and when there are being irradiated while GSH with NIR light, and the rate of release of drug can be into
One step increases, and illustrates that GSH and NIR irradiation has synergistic effect to the release for accelerating drug, related data is shown in Figure 10.
Embodiment 14
The 4T1 cell of logarithmic growth phase adjusts concentration of cell suspension, is inoculated in 96 orifice plates, and 100ul is added in every hole,
It is placed in incubator.After cell is adherent, it is separately added into the DOX solution of serial various concentration, MSN-SS-Cy/TPGS blank carries
Body and MSN-SS-Cy/TPGS medicine-carried system, wherein MSN-SS-Cy/TPGS empty vectors and MSN-SS-Cy/TPGS medicine-carried system
And with the NIR laser 2W power illumination 1min of 808nm.Continue after being incubated for for 24 hours, 50 μ L of 2mg/mL MTT solution is added, continues
Culture solution in hole is carefully sucked after being incubated for 4h, every hole is added 150 μ l dimethyl sulfoxides, sets low-speed oscillation 10min on shaking table, make to tie
Brilliant object sufficiently dissolves.The light absorption value A in each hole is measured at microplate reader 570nm, and calculates the relative survival rate of cell.
The result shows that MSN-SS-Cy/TPGS empty vectors do not have the survival rate of cell within the scope of test concentrations substantially
It influences, cell survival rate maintains always 80% or more, it is believed that blank nanoparticle prepared by the present invention is without obvious thin
Cellular toxicity.And under NIR light irradiation, the temperature of carrier increases, and certain toxic side effect can be generated to cell.For MSN-SS-
For Cy/TPGS medicine-carried system under the NIR light irradiation of 808nm, the inhibiting rate highest of cell, toxicity is maximum, illustrates the load medicine of building
The synergy of chemotherapy and thermotherapy may be implemented in system, and related data is shown in attached drawing 11.
Claims (10)
1. a kind of mesoporous silicon oxide drug delivery system, which is characterized in that TPGS is coated on disulfide bond grafting Cypate's
Mesoporous silicon dioxide nano particle and be made.
2. mesoporous silicon oxide drug delivery system according to claim 1, which is characterized in that utilize disulfide bond will
TPGS is coated on outermost layer in meso-porous titanium dioxide silicon face, and using film hydration method by Cypate covalence graft.
3. the preparation method of mesoporous silicon oxide drug delivery system according to claim 1, which is characterized in that
1) preparation of mesoporous silicon dioxide nano particle:
2) preparation of sulfhydrylation mesoporous silicon dioxide nano particle:
3)MSN-SS-NH2Preparation:
4) preparation of MSN-SS-Cy:
5) preparation of MSN-SS-Cy/TPGS.
4. preparation method according to claim 3, which is characterized in that step carries mesoporous silicon dioxide nano particle in (2)
Body is distributed in dehydrated alcohol, and under nitrogen protection with (3- mercaptopropyi) trimethoxy silane back flow reaction, sulfhydrylation is made
Then the mesoporous silicon carrier of modification removes cetyl trimethylammonium bromide template using acid extraction method, mesoporous by having exposed
The mesoporous silicon dioxide nano particle in duct is as pharmaceutical carrier.
5. preparation method according to claim 3, which is characterized in that step receives sulfhydrylation mesoporous silicon oxide in (3)
Nanoparticle carrier and S- (2- aminoethanethio) -2- thiopyridine (Py-SS-NH2) back flow reaction, two sulphur are introduced in mesoporous silicon face
MSN carrier (the MSN-SS-NH that amino terminal is connected by disulfide bond is made in key2)。
6. preparation method according to claim 3, which is characterized in that Cypate is dissolved in dimethyl sulfoxide by step (4)
In, 1- ethyl-(3- dimethylaminopropyl) carbodiimide hydrochloride and n-hydroxysuccinimide activation one hour is being added
Afterwards, with MSN-SS-NH2The MSN carrier (MSN-SS-Cy) that Cypate is grafted by disulfide bond is prepared in carrier reflux.
7. preparation method according to claim 3, which is characterized in that utilize the film hydration method packet of improvement in step (5)
It is covered with TPGS, the MSN-SS-Cy of surface cladding TPGS is made, the density of the TPGS is (1.00-1.10) * 103kg/m3。
8. mesoporous silicon oxide drug delivery system described in claim 1 has thermotherapy and reduction response type drug release energy in preparation
Application in the drug of power.
9. application as claimed in claim 8, which is characterized in that the drug is adriamycin, camptothecine or taxol.
10. application as claimed in claim 8 or 9, which is characterized in that drug is loaded into the sulfydryl of claim 3 step (2)
Change mesoporous silicon dioxide nano particle, is then carried according still further to the MSN-SS-Cy/TPGS nanometer of step (3), (4), (5) preparation drug containing
The mass ratio of body, drug and MSN-SS-Cy carrier is between 1:1 to 1:5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711151069.4A CN109806394B (en) | 2017-11-18 | 2017-11-18 | Mesoporous silica drug delivery system and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711151069.4A CN109806394B (en) | 2017-11-18 | 2017-11-18 | Mesoporous silica drug delivery system and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109806394A true CN109806394A (en) | 2019-05-28 |
| CN109806394B CN109806394B (en) | 2021-09-24 |
Family
ID=66598013
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711151069.4A Active CN109806394B (en) | 2017-11-18 | 2017-11-18 | Mesoporous silica drug delivery system and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109806394B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113144216A (en) * | 2021-03-23 | 2021-07-23 | 昆明理工大学 | ICG-MSNs nano material and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102949728A (en) * | 2012-12-12 | 2013-03-06 | 重庆大学 | Meso-porous silicon nano-drug carrier with both reduction responsiveness and targeting ability and preparation method thereof |
| EP2762488A2 (en) * | 2011-09-30 | 2014-08-06 | Korea University Research and Business Foundation | Peptide for synthesizing silica, and use thereof |
| CN105056233A (en) * | 2015-08-12 | 2015-11-18 | 苏州大学 | Multifunctional mesoporous silica nanoparticles having near-infrared photothermal and in-vivo fluorescence imaging characteristics as well as preparation method and application of mesoporous silica nanoparticles |
-
2017
- 2017-11-18 CN CN201711151069.4A patent/CN109806394B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2762488A2 (en) * | 2011-09-30 | 2014-08-06 | Korea University Research and Business Foundation | Peptide for synthesizing silica, and use thereof |
| CN102949728A (en) * | 2012-12-12 | 2013-03-06 | 重庆大学 | Meso-porous silicon nano-drug carrier with both reduction responsiveness and targeting ability and preparation method thereof |
| CN105056233A (en) * | 2015-08-12 | 2015-11-18 | 苏州大学 | Multifunctional mesoporous silica nanoparticles having near-infrared photothermal and in-vivo fluorescence imaging characteristics as well as preparation method and application of mesoporous silica nanoparticles |
Non-Patent Citations (3)
| Title |
|---|
| QINFU ZHAO等: "Dual-stimuli responsive hyaluronic acid-conjugated mesoporous silica for targeted delivery to CD44-overexpressing cancer cells", 《ACTA BIOMATERIALIA》 * |
| 侯仲等: "还原响应型介孔二氧化硅-巯基嘌呤共价载药系统的构建与初步评价", 《沈阳药科大学学报》 * |
| 黄丽: "Cypate/DOX-SiO2纳米粒用于肿瘤诊疗的初步研究", 《中国优秀硕士学位论文全文数据库》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113144216A (en) * | 2021-03-23 | 2021-07-23 | 昆明理工大学 | ICG-MSNs nano material and preparation method and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109806394B (en) | 2021-09-24 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zheng et al. | Biodegradable hypocrellin derivative nanovesicle as a near-infrared light-driven theranostic for dually photoactive cancer imaging and therapy | |
| CN108354901A (en) | For chemotherapy of tumors and the pH/ of photo-thermal combination therapy reduction Dual Sensitive multifunctional nano micellas and its application | |
| CN106139144A (en) | A kind of hyaluronic acid decorated golden Nano carbon balls with synergistic antitumor characteristic and preparation method and application | |
| WO2010040312A1 (en) | Composite material and its prepration, using in tumor therapy and aititumor medicine | |
| CN105056233A (en) | Multifunctional mesoporous silica nanoparticles having near-infrared photothermal and in-vivo fluorescence imaging characteristics as well as preparation method and application of mesoporous silica nanoparticles | |
| CN111760024B (en) | Permeation enhanced gold nanocluster drug-loaded targeting preparation and preparation method and application thereof | |
| Zhang et al. | Killing three birds with one stone: Near-infrared light triggered nitric oxide release for enhanced photodynamic and anti-inflammatory therapy in refractory keratitis | |
| CN106512002B (en) | Multifunctional nano hybrid integrating CT imaging and phototherapy and preparation method thereof | |
| CN109248327B (en) | Mesoporous silica drug delivery system and application thereof | |
| CN108324955B (en) | Preparation method of ultra-small copper sulfide loaded hollow mesoporous silicon targeted nano drug-loaded compound | |
| CN106519213B (en) | A kind of platinum two azoles of fluorine boron and preparation method and application | |
| CN113559064B (en) | Novel self-oxygen-supply liposome nanoparticle and preparation method and application thereof | |
| CN104940945A (en) | Hollow mesoporous copper sulfide compound modified by hyaluronic acid and preparation method and application thereof | |
| CN105126113A (en) | Preparation method and application of transferrin modified hollow mesoporous copper sulfide/artesunate nanoparticles | |
| WO2021083370A1 (en) | Preparation and use of nanomaterial specifically activating immune system | |
| CN106166141A (en) | A kind of Multifunctional composite nanometer medicine for tumor imaging and treatment and preparation method thereof | |
| CN105194679A (en) | Preparation method and application of titanium dioxide-graphene oxide composite material modified by hyaluronic acid of antitumor drug nanometer layer | |
| CN112933229B (en) | Carrier-free self-assembly nanoparticle of IR820 and atovaquone and preparation method and application thereof | |
| CN110448699A (en) | The neoplastic cell nuclei targeted medicament carrying nano particle and preparation method of seven methine Hua Jingsu class dyestuffs are modified comprising functional polypeptide | |
| Wu et al. | Composite phospholipid-coated hollow mesoporous silica nanoplatform with multi-stimuli responsiveness for combined chemo-photothermal therapy | |
| CN109806394A (en) | A kind of mesoporous silicon oxide drug delivery system and its application | |
| CN104707138B (en) | A kind of preparation method of the three-layer nuclear shell structure pharmaceutical carrier near infrared light remote response | |
| CN106606783A (en) | Drug delivery system for targeting co-delivery of photosensitizer and chemotherapeutic drug | |
| CN105949344B (en) | A kind of cation hyperbranched polysaccharide derivates and its in enhancing blood porphyrin photosensitizer to the application in terms of tumour cell phototoxicity | |
| CN113134084B (en) | Glutathione stimulated induced aggregation type composite gold nanoparticle and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |