CN104188910A - Targeting light-operated nitric oxide release nanometer composite material medicine system and preparation method thereof - Google Patents

Targeting light-operated nitric oxide release nanometer composite material medicine system and preparation method thereof Download PDF

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CN104188910A
CN104188910A CN201410280092.3A CN201410280092A CN104188910A CN 104188910 A CN104188910 A CN 104188910A CN 201410280092 A CN201410280092 A CN 201410280092A CN 104188910 A CN104188910 A CN 104188910A
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carrier
tpy
drug system
exogenous
donor
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CN104188910B (en
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刘劲刚
向慧静
唐伟伟
邓乔
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East China University of Science and Technology
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East China University of Science and Technology
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Abstract

The invention provides a preparation method for a multifunctional nanometer medicine system with the functions of fluorescence tracing, targeting conveying and intracellular light-operated release of nitric oxide. The nanometer system takes titanium dioxide nanoparticle, carbon quantum dot, graphene quantum dot or the like as a carrier, takes metal ruthenium and manganese nitrosyl compounds as exogenous NO donors, and takes folic acid, galactose molecules and the like as targeting guiding groups. The nitric oxide release nanometer system has good biological compatibility and stability, has fluorescence tracing function, is capable of performing nitric oxide selective light-operated release on specific cancer cells, and has potential application prospect and commercial business in the fields of treatment, photodynamic therapy (PDT) and the like on diseases caused by lack of NO in body.

Description

The light-operated release nitric oxide of targeting nano composite material drug system and preparation method thereof
Technical field
The present invention relates to inorganic nano drug world.Concrete, the invention provides a class and there is nitric oxide production multifunctional nanocomposites drug system of light-operated delivery and preparation method thereof in fluorescent tracing, targeted, cell.
Background technology
Nitric oxide (NO) is a kind of endogenic biological micromolecule, has very important effect in a lot of physiological process, as regulated blood pressure, immunity and inflammatory reaction, nerve conduction and apoptosis etc.Its site discharging in vivo, time and dosage are depended in the performance of NO biological function.Low concentration (nmolL -1) NO promote cell enlargement, and high concentration (μ molL -1-mmolL -1) can cause the apoptosis of cell.NO is a kind of hydroxyl radical gas, easy and oxygen reaction, and extremely unstable, the half-life is short.Although NO has many biological regulation functions, due to its unstability, limit NO in clinical medical application.Therefore, research worker wishes to find a kind of ectogenic NO donor, and it can stablize preservation between the storage life, and when needed by extraneous stimulation, can discharge in specific position the NO of suitable dose, thereby reaches the therapeutical effect of NO mediation.
People comprise that to all kinds of ectogenic NO donors organic NO donor and metal nitrosyl (M-NO) donor have carried out large quantity research (Huerta, S.et al.Intl.J.Oncology2008,33,909 – 927; Sortino, S.Chem.Soc.Rev.2010,39,2903 – 2913; Fry, N.L.et al.Acc.Chem.Res.2011,44,289-298; Naghavi, N., et al.Small2013,9,22 – 35; Kim, J.et al.J.Mater.Chem.B2014,2,341 – 356.).All kinds of delivery platforms of organic NO donor and composition thereof, as organic nitrates RONO 2, nitrites RONO, nitrosothiols RSNO etc., although in certain external condition (illumination, pH value changes, variations in temperature etc.) stimulate under can discharge NO, but such donor system is generally unstable in vivo, the dosage of instant-free NO is uncontrollable, and lacks targeted function.And the shortage of targeting often causes NO to kill and wound Normocellular, thereby bring very large toxic and side effects.
Light, as a kind of Noninvasive easy-regulating means, can accurately be controlled the dosage of metal nitrosyl donor release NO by capable of regulating illumination intensity and light application time.The metal nitrosyl donor of research mainly contains metal M n, Fe, Cr, the nitrosyl compounds such as Ru at present.The metal nitrosyl donor of early stage research can only discharge NO under the exciting of high-octane ultraviolet light, so research worker is by changing the part of metal and being connected some chromophores (as fluorescein, resorufin, red sulphonyl) or being loaded on the surface (go up converting material, quantum dot) of some functional materials and then can discharge NO under visible ray, near infrared light, be conducive to like this application (Rose of these compounds in living things system, M.J.et al.Coord.Chem.Rev.2008,252,2093 – 2114; Tfouni, E.et al.Coord.Chem.Rev.2010,254,353 – 371; Ford, P.C.Nitric Oxide2013,34,56 – 64.).Above-mentioned metal nitrosyl donor shines to control by ambient light can realize NO release in vivo, but how can in specific region or specific cell, be that selectivity discharges the still Challenge of NO that is applicable to concentration.
Therefore, study a kind of medicine that can fix a point, regularly, quantitatively discharge NO (or system) particularly important.
Summary of the invention
The object of this invention is to provide a kind ofly there is targeting guiding, visible ray controlled release is put nano composite material drug system of NO and preparation method thereof.
A first aspect of the present invention, provides a kind of nano composite material drug system, and described drug system comprises:
Carrier, described carrier is nanoparticle;
The exogenous NO gas donor being linked with described carrier; With
The targeting homing device being linked with described carrier.
In another preference, described carrier is selected from lower group: titanium dioxide nano-particle, carbon quantum dot, graphene quantum dot, up-conversion nanoparticles, magnetic nano-particle or its combination.
In another preference, described carrier is the carrier of surface amination.
In another preference, described exogenous NO gas donor is metal nitrosyl compound; Preferably, described metal nitrosyl compound has the structure shown in following formula I:
[(tpy′)M(R 1)(NO)](PF 6) 3 (I)
Wherein, tpy ' is three tooth containing n-donor ligands, is preferably selected from: 4'-formic acid-2,2':6', 2 "-terpyridyl or derivatives thereof, 2,2':6', 2 "-terpyridyl or derivatives thereof or its combination;
R 1for bidentate containing n-donor ligand, be preferably selected from: DAMBO (boron two pyrroles's methyl-derivatives), o-phenylenediamine analog derivative, diaminourea fluorescein derivative, diaminourea Rhodamine Derivatives or 3-formic acid-o-phenylenediamine or its combination;
M is metal.
In another preference, tpy ' and R 1in at least one contains hydroxy-acid group.
In another preference, M is selected from lower group: Ru, Mn.
In another preference, described metal nitrosyl compound is selected from lower group: metal Ru nitrosyl compound, or manganese metal nitrosyl compound.
In another preference, described metal Ru nitrosyl compound is [(tpy ') Ru (R 1) (NO)] (PF 6) 3, wherein tpy ' is 4'-formic acid-2,2':6', 2 " and-terpyridyl, R 1for DAMBO and/or o-phenylenediamine.
In another preference, described manganese metal nitrosyl compound is [(tpy ') Mn (R 1) (NO)] (PF 6) 3, wherein tpy ' is 4'-formic acid-2,2':6', 2 " and-terpyridyl, R 1for DAMBO and/or o-phenylenediamine.
In another preference, described targeting homing device is selected from lower group: folate molecule, galactose molecule, biotin or its combination.
In another preference, described carrier is linked by covalent bond and described exogenous NO gas donor; And/or
In another preference, described carrier is linked by covalent bond and described targeting homing device.
In another preference, the radius of described nano composite material drug system is 3-200nm, is preferably 5-100nm.
In a second aspect of the present invention, a kind of method of preparing the nano composite material drug system described in first aspect present invention is provided, described method comprises step:
(1) provide exogenous NO gas donor, targeting homing device and carrier nanoparticle;
(2) described exogenous NO gas donor, described targeting homing device and described carrier nanoparticle are carried out to covalency load, thereby form the drug system described in first aspect present invention.
In another preference, in step (1), described exogenous NO gas donor comprises metal nitrosyl compound [(tpy ') M (R 1) (NO)] (PF 6) 3, tpy ' is 4'-formic acid-2,2':6', 2 " and-terpyridyl, M is metal Ru (Ru) or manganese metal (Mn), R 1for DAMBO (boron two pyrroles's methyl-derivatives) or o-phenylenediamine.
In another preference, described carrier nanoparticle is the nanoparticle of surface amination.
In another preference; in described step (2); first described targeting homing device and described surface amination carrier nanoparticle are carried out to amidation process; there is the nanoparticulate carriers of targeting homing device to carry out amidation process described metal nitrosyl compound and described load again, thereby make described nano composite material drug system.
In another preference, described exogenous NO gas donor comprises metal nitrosyl compound.
In another preference, described method also comprises the following steps of the described exogenous NO gas donor of preparation before in step (1):
(a1) provide formula A compound,
M(tpy′)Cl 3 (A)
Wherein, M is metal, is preferably selected from lower group: Ru, Mn;
Tpy ' is three tooth containing n-donor ligands;
(a2) in inert atmosphere, with described M (tpy ') Cl 3with R 1and NH 4pF 6reaction, forms [(tpy ') M (R 1) Cl] (PF 6) 3; Wherein, R 1be selected from lower group: DAMBO (boron two pyrroles's methyl-derivatives) or o-phenylenediamine;
(a3) in suitable solvent, by described [(tpy ') M (R 1) Cl] (PF 6) 3react with nitrite, obtain [(tpy ') M (R 1) NO 2] (PF 6);
(a4) by [(tpy ') M (R 1) NO 2] (PF 6) and acid and NH 4pF 6reaction, obtains described exogenous NO gas donor.
In another preference, acid is selected from following group: HNO 3, H 2sO 4, HPF 6or hydrochloric acid.
In another preference, described step (a2) is carried out in inert gas environment.
In another preference, described step (a4) is carried out under 0 DEG C~room temperature (as 25 DEG C).
In another preference, described method also comprises in step (1) following steps of preparing nanoparticulate carriers before:
By (4-NH 2)-C 6h 4-PO 3h reacts with nanoparticle, obtains the nanoparticulate carriers of surface amination.
In another preference, nanoparticle is dispersed in the aqueous solution of pH9.
In another preference, in step (2), described nanoparticle is first reacted with targeting homing device, and then supply precursor reactant with exogenous NO gas, form described nano composite material drug system.
In another preference, the mol ratio of described targeting homing device and described exogenous NO gas donor is (1 ± 0.2): (8 ± 1.6), preferably, are (1 ± 0.5): (8 ± 4).
In another preference, in described step (2), under existing, coupling agent carries out, preferably, described coupling agent is selected from lower group: EDC/NHS, wherein EDC is 1-ethyl-3-[3-dimethylaminopropyl] carbodiimides hydrochloride, NHS is N-hydroxy-succinamide.
In a third aspect of the present invention, the nano composite material drug system described in a kind of first aspect present invention is provided, and pharmaceutically acceptable carrier.
In another preference, described pharmaceutical composition also comprises other medicines as active component.
In a fourth aspect of the present invention, provide the purposes of the nano composite material drug system described in a kind of first aspect present invention.For the preparation of medicine, described medicine is targeted drug and has visible ray controlled release NO.
In another preference, described medicine is for regulating and controlling or regulating the physiological process that is selected from lower group: blood pressure, immunity, inflammation, nerve conduction or apoptosis.
In a fifth aspect of the present invention, the method for a kind of release nitric oxide (NO) is provided, by the nano composite material drug system described in radiation of visible light first aspect present invention, thereby make described nano composite material drug system discharge NO.
In another preference, described visible light wavelength is 400-800nm.
In another preference, described method is non-therapeutic and non-diagnostic.
In another preference, described release is in vitro, and nano composite material drug system in solution is carried out to illumination, thereby discharges NO.
In another preference, radiation modality is pulse irradiation or prolonged exposure.
In another preference, the light intensity of irradiation is 10-1000 milliwatt/square centimeter.
In a sixth aspect of the present invention, a kind of method to cell by nano composite material drug system targeted of non-therapeutic is provided, comprise step: the nano composite material drug system described in first aspect present invention is hatched together with described cell, the surface receptor or the surface protein that match with described targeting homing device are carried in the surface of wherein said cell, thereby make described nano composite material drug system by extremely described cell of targeted.
In another preference, described cell is the HeLa cell of folacin receptor overexpression.
In another preference, targeted cells is the MCF-7 cell of folic acid weak expression.
In another preference, described incubation conditions is 37 ± 2 DEG C (preferably 37 DEG C), 2 ± 1 hours, and the concentration of described nano composite material drug system is 5-500 ug/ml.
In another preference, described method also comprises the release conditions with NO probe in detecting NO.
In another preference, NO probe is DAF-FM DA.
In another preference, targeted cells is hatched 1-24 hour with nano composite material drug system together with NO probe, and then with radiation of visible light 0.5-30 minute.
In a seventh aspect of the present invention, the method for NO of conveying a kind of is provided, comprise step: use the nano composite material drug system described in first aspect present invention to the object of needs.
In another preference, described object comprises mammal (for example people).
In another preference, described method also comprises irradiates with visible ray, thus light-operated release NO.
In should be understood that within the scope of the present invention, above-mentioned each technical characterictic of the present invention and can combining mutually between specifically described each technical characterictic in below (eg embodiment), thus form new or preferred technical scheme.As space is limited, tire out and state no longer one by one at this.
Brief description of the drawings
Fig. 1 has shown nitric oxide nano composite material drug system structural representation.This system is taking nanoparticle as carrier, and carrier cochain is connected to exogenous NO gas donor, and on carrier, chain is connected to targeting homing device simultaneously.
Fig. 2 has shown { Ru-NO@TiO 2the sign of Nano medication system.Wherein figure (A) is { Ru-NO@TiO 2the transmission electron microscope picture of NPs} and particle size distribution; Figure (B) is { Ru-NO@TiO 2diffuse-reflectance uv-vis spectra and the infrared spectrum of NPs}.
Fig. 3 has shown { Ru-NO@TiO 2the illumination in saline solution of NPs} Nano medication system discharges NO.Wherein figure (A) is visible light pulses irradiation.{ Ru-NO@TiO 2nPs} concentration: 2.0 mg/ml; Light intensity: 100 milliwatt/square centimeters, λ >400nm.Figure (B) is visible ray prolonged exposure.{ Ru-NO@TiO 2nPs} concentration: 1.0 mg/ml; Light intensity: 300 milliwatt/square centimeters, λ >400nm.
Fig. 4 has shown { Ru-NO@TiO 2the targeted cells of NPs} Nano medication system is carried.Wherein figure (A) is the HeLa cell and { Ru-NO@TiO of folacin receptor overexpression 2nPs} (50.0 ug/ml) is in 37 DEG C of laser co-focusing figure of hatching after 2 hours; The MCF-7 cell and { Ru-NO@TiO of figure (B) folacin receptor weak expression 2nPs} (50.0 ug/ml) is in 37 DEG C of laser co-focusing figure of hatching after 2 hours.
Fig. 5 has shown { Ru-NO@TiO 2in the targeted cells of NPs} Nano medication system, NO delivers.Wherein figure (A) is the HeLa cell and { Ru-NO@TiO of folacin receptor overexpression 2nPs} (50 μ g/mL) and NO probe DAF-FM DA (5 μ M) cultivate the laser co-focusing figure after 6 hours together.Figure (B) is the HeLa cell and { Ru-NO@TiO of folacin receptor overexpression 2nPs} (50 μ g/mL) and NO probe DAF-FM DA (5 μ M) cultivate 6 hours together, then use the laser co-focusing figure of radiation of visible light after 1 minute.(light intensity: 200 milliwatt/square centimeters, λ >400nm).
Detailed description of the invention
The inventor is through extensive and deep research, by a large amount of screening and tests, made first the light-operated release of a kind of novel targeting NO nano composite material drug system, described system comprises carrier and is linked exogenous NO gas donor and the targeting homing device of (as covalently bound) with described carrier.System of the present invention not only can discharge rapidly NO molecule under radiation of visible light, and can carry out within a large range by regulating illumination time and intensity of illumination (nM~μ M) adjustment release NO concentration.In addition, nano composite material drug system of the present invention also has targeting, the good advantage such as bio-compatibility and stability.Complete on this basis the present invention.
Term
As used herein, term " drug system of the present invention ", " system of the present invention ", " the light-operated release of targeting NO nano composite material drug system ", " the light-operated release of targeting NO nano composite material medicine " and " the light-operated release of targeting NO nano composite material pharmaceutical composition " are used interchangeably, all refer to taking nanoparticle as carrier, carrier cochain is connected to exogenous NO gas donor, and on carrier, while chain is connected to the compositions of targeting homing device.
Nanoparticulate carriers
Being applicable to nanoparticulate carriers of the present invention and being not particularly limited, can be the conventional various nanoparticulate carriers in this area.Representational example includes, but are not limited to: titanium dioxide nano-particle, carbon quantum dot, graphene quantum dot, up-conversion nanoparticles, magnetic nano-particle or its combination.
Typically, the particle diameter of nanoparticulate carriers is 3-200nm, is preferably 5-100nm.
The carrier that the preferred nanoparticulate carriers of one class is surface amination.
Exogenous NO gas donor
Being applicable to exogenous NO gas donor of the present invention and being not particularly limited, can be the conventional various exogenous NO gas donors in this area.Representational example includes, but are not limited to: metal nitrosyl compound.
Typically, exogenous NO gas donor is metal Ru nitrosyl compound or manganese metal nitrosyl compound.
In the present invention, the weight ratio of exogenous NO gas donor and nanoparticulate carriers is 0.8-10:100.
Targeting homing device
Being applicable to targeting homing device of the present invention and being not particularly limited, can be the conventional various targeting homing devices in this area.Representational example includes, but are not limited to: folate molecule, galactose molecule, biotin or its combination.
In the present invention, the mol ratio of described targeting homing device and described exogenous NO gas donor is not particularly limited, typically the mol ratio of described targeting homing device and described exogenous NO gas donor is (1 ± 0.2): (8 ± 1.6) are more preferably 1:8.
Composite drug system
Composite drug system of the present invention all refers to that, taking nanoparticle as carrier, carrier cochain is connected to exogenous NO gas donor, and on carrier, while chain is connected to the composite particle of targeting homing device.A kind of schematic diagram of simplification as shown in Figure 1.
Composite drug system of the present invention can be used for carrying NO to cell, tissue or object, thereby for regulating or regulation and control physiological process, the especially physiological process relevant to NO.
In another preference, described physiological process is selected from lower group: blood pressure, immunity, inflammation, nerve conduction or apoptosis etc.
Preparation method
The present invention also provides the preparation method of drug system of the present invention, generally includes following steps:
(1) provide exogenous NO gas donor, targeting homing device and carrier nanoparticle;
(2) described exogenous NO gas donor, described targeting homing device and described carrier nanoparticle are carried out to covalency load, thereby form described NO nano composite material drug system.
In another preference, in step (1), described exogenous NO gas donor comprises metal nitrosyl compound [(tpy ') M (R 1) (NO)] (PF 6) 3, tpy ' is 4'-formic acid-2,2':6', 2 " and-terpyridyl, M is metal Ru (Ru) or manganese metal (Mn), R 1for DAMBO (boron two pyrroles's methyl-derivatives) or o-phenylenediamine.
In another preference, described carrier nanoparticle is the nanoparticle of surface amination.
In another preference, described targeting homing device is selected from: folate molecule, galactose molecule, biotin or its combination.
In another preference; in described step (2); first described targeting homing device and described surface amination carrier nanoparticle are carried out to amidation process; there is the nanoparticulate carriers of targeting homing device to carry out amidation process described metal nitrosyl compound and described load again, thereby make described nano composite material drug system.
Major advantage of the present invention comprises:
(1) provide a kind of and can discharge rapidly NO molecule under radiation of visible light, and can carry out by regulating illumination time and intensity of illumination the nano composite material drug system of adjustment release NO concentration (nM~μ M).
(2) nano composite material drug system of the present invention has blue autofluorescence, can be in cell the endocytosis situation of spike drug system, and the NO discharging in cell can detect by NO fluorescent probe (DAF-FM-DA).
(3) nano composite material drug system of the present invention can be optionally carried (as HeLa cell) to having in the tumor cell of folacin receptor (FR) overexpression.
(4) nano composite material drug system of the present invention has good bio-compatibility and stability under dark condition.
Below in conjunction with specific embodiment, further set forth the present invention.Should be understood that these embodiment are only not used in and limit the scope of the invention for the present invention is described.The experimental technique of unreceipted actual conditions in the following example, conventionally according to normal condition, or the condition of advising according to manufacturer.Unless otherwise indicated, otherwise percentage ratio and umber are percentage by weight and parts by weight.
The nitrosyl compound [(tpy of embodiment 1 metal Ru cOOH) Ru (DAMBO) (NO)] (PF 6) 3synthetic
(1) [(tpy cOOH) Ru (DAMBO) (Cl)] (PF 6) synthetic (tpy cOOHfor 4'-formic acid-2,2':6', 2 "-terpyridyl):
In the there-necked flask of 100mL, add Ru (tpy cOOH) Cl 3(150mg, 0.31mmol), DAMBO (117mg, 0.33mmol), LiCl (5mg, 2.0mmol) and Et 3n0.4mL, evacuation and logical nitrogen are distinguished three times, add the EtOH/H of 40mL 2o (3:1, v/v), at N 2in atmosphere, reflux 8 hours, sucking filtration while hot, the dark red filtrate obtaining is concentrated to several milliliters, to be cooled after room temperature, adds excessive saturated NH 4pF 6solution, is placed on this mixture the refrigerator overnight of 5 DEG C.Filter out red-brown precipitation, use H 2o and Et 2o washs respectively three times, vacuum drying.Obtain target product 182.3mg, productive rate is 64%.
(2) [(tpy cOOH) Ru (DAMBO) (NO 2)] (PF 6) synthetic:
By [(tpy cOOH) Ru (DAMBO) (Cl)] (PF 6) (100mg, 0.11mmol) and excessive AgNO 3(37mg, 0.22mmol) joins in the round-bottomed flask of 100mL, adds 30mL CH 3cN-H 2o (1:1, v/v), reflux 2 hours, solution colour becomes purple gradually by redness, by this mixture cool to room temperature, filters out linen AgCl, in filtrate, adds excessive NaNO 2(103.5mg, 1.5mmol), reflux 6 hours, treats that solution is cooled to room temperature, and solution is concentrated to several milliliters, adds excessive saturated NH 4pF 6solution, is placed on this mixture the refrigerator overnight of 5 DEG C.Filter out red-brown precipitation, use H 2o and Et 2o washs respectively three times, vacuum drying.Obtain target product 78.2mg, productive rate is 77%.
(3) [(tpy cOOH) Ru (DAMBO) (NO)] (PF 6) 3synthetic:
At 273K temperature, by [(tpyCOOH) Ru (DAMBO) (NO 2)] (PF 6) (100mg, 0.11mmol) join in the round-bottomed flask of 25mL, then dropwise drips 2mL HNO 3(2molL -1) in above-mentioned solid, form the solid of pasty state, stir after 30 minutes, add excessive saturated NH 4pF 6solution, is placed on this mixture the refrigerator overnight of 5 DEG C.Filter out red-brown precipitation, use H 2o and Et 2o washs respectively three times, vacuum drying.Obtain target product 75.8mg, productive rate is 58%.
{ the Ru-NO@TiO of the present embodiment 2the transmission electron microscope picture of Nano medication system and particle size distribution as shown in Figure 2 A, diffuse-reflectance uv-vis spectra and the results of FT-IR are as Fig. 2 B.
Embodiment 2 nano composition drug system { Ru-NO@TiO 2synthetic
(1) NH 2@TiO 2(the TiO of surface amination 2nanoparticle) preparation
In the round-bottomed flask of 50mL, add 9.3mg (4-NH 2)-C 6h 4-PO 3after H, add the aqueous solution of 2mL pH9, after dissolution of solid, add 18mL TiO 2aqueous dispersion in above-mentioned solution, mixed liquor at room temperature stirs and spends the night, the dispersion liquid obtaining turns lower centrifugal 10 minutes 1000, removes supernatant, washes nanoparticle twice with water, the TiO of the surface amination obtaining 2nanoparticle, is dispersed in aqueous solution again, for subsequent use.
(2) nano composition drug system { Ru-NO@TiO 2preparation
By [(tpy cOOH) Ru (DAMBO) (NO)] (PF 6) 3(100mg, 0.08mmol) and FA (folic acid) (5.0mg, 0.01mmol) are dissolved in the DMF of 5mL, add EDC/NHS activation 30 minutes, afterwards, add 50.0mg NH 2@TiO 2nPs, reacts 12 hours.Centrifugal, remove supernatant, with DMF and H 2o washs respectively three times, finally obtains { Ru-NO@TiO 2nPs}, is scattered in water again.
Embodiment 3{Ru-NO@TiO 2nPs} Nano medication system pulsed light in saline solution shines and discharges NO
{ Ru-NO@TiO 2the concentration of NPs}: 2.0mg/mL;
Light intensity: 100 milliwatt/square centimeters, λ >400nm.
Visible light pulses (5 seconds to 25 seconds) by different time is irradiated sample, and measurement result is as shown in A in Fig. 3.
Result: the nitric oxide concentration detecting in solution increases progressively (in Fig. 3 shown in A) with the burst length.This shows, can obtain instant required suitable nitric oxide concentration by the regulating impulse time.
Embodiment 4{Ru-NO@TiO 2nPs} Nano medication system continuous light in saline solution discharges NO
{ Ru-NO@TiO 2the concentration of NPs}: 1.0mg/mL;
Light intensity: 300 milliwatt/square centimeters, λ >400nm.
Can obtain the nitric oxide solution of about 9 micromoles per liter by continuing the sample of radiation of visible light 1.0mg/mL, in the prolonged exposure of 2-3 hour, this concentration remain unchanged (in Fig. 3 shown in B).
Embodiment 5{Ru-NO@TiO 2the targeted cells of NPs} Nano medication system is carried
By the HeLa cell and { Ru-NO@TiO of folacin receptor overexpression 2nPs} (50.0 μ g/mL) is hatched in 37 DEG C, carries out laser co-focusing observation after 2 hours.
Result: can be observed obvious blue-fluorescence in HeLa cell, this result shows in a large number { Ru-NO@TiO 2nPs} nanometer system has entered cell, and major part is distributed in Cytoplasm, also has distribution (in Fig. 4 shown in A) in nucleus.If extend the cultivation time to 8 hour, can be observed and enter nuclear amount showed increased.
Embodiment 6{Ru-NO@TiO 2the targeted cells of NPs} Nano medication system is carried
By the MCF-7 cell and { Ru-NO@TiO of folacin receptor weak expression 2nPs} (50.0 μ g/mL) is hatched in 37 DEG C, carries out laser co-focusing observation after 2 hours.
Result: only observe faint blue-fluorescence in MCF-7 cell, this result shows { Ru-NO@TiO 2nPs} nanometer system enters the amount of MCF-7 cell less (in Fig. 4 shown in B).
Embodiment 7{Ru-NO@TiO 2in the targeted cells of NPs} Nano medication system, NO delivers
By the HeLa cell and { Ru-NO@TiO of folacin receptor overexpression 2nPs} (50 μ g/mL) and NO probe DAF-FM DA (5 μ M) cultivate together, after 6 hours, carry out laser co-focusing observation.
Result: do not having under rayed condition, only observing NO probe and show the faint green fluorescence of probe self (in Fig. 5 shown in A) in cell.
Embodiment 8{Ru-NO@TiO 2in the targeted cells of NPs} Nano medication system, NO delivers
By the HeLa cell and { Ru-NO@TiO of folacin receptor overexpression 2nPs} (50 μ g/mL) and NO probe DAF-FM DA (5 μ M) cultivate 6 hours together, then use radiation of visible light after 1 minute, carry out laser co-focusing observation.(light intensity: 200 milliwatt/square centimeters, λ >400nm).
Result: under radiation of visible light condition, observe NO probe and show very strong green fluorescence (in Fig. 5 shown in B) in cell, this result shows that the nitric oxide that the irradiation by controlling light can realize in targeted cells discharges.
All documents of mentioning in the present invention are all quoted as a reference in this application, are just quoted separately as a reference as each section of document.In addition should be understood that those skilled in the art can make various changes or modifications the present invention after having read above-mentioned teachings of the present invention, these equivalent form of values fall within the application's appended claims limited range equally.

Claims (10)

1. a nano composite material drug system, is characterized in that, described drug system comprises:
Carrier, described carrier is nanoparticle;
The exogenous NO gas donor being linked with described carrier; With
The targeting homing device being linked with described carrier.
2. drug system as claimed in claim 1, described carrier is selected from lower group: titanium dioxide nano-particle, carbon quantum dot, graphene quantum dot, up-conversion nanoparticles, magnetic nano-particle or its combination; And/or
Described carrier is the carrier of surface amination.
3. drug system as claimed in claim 1, described exogenous NO gas donor is metal nitrosyl compound; Preferably, described metal nitrosyl compound has the structure shown in following formula I:
[(tpy′)M(R 1)(NO)](PF 6) 3 (I)
Wherein, tpy ' is three tooth containing n-donor ligands, is preferably selected from: 4'-formic acid-2,2':6', 2 "-terpyridyl or derivatives thereof, 2,2':6', 2 "-terpyridyl or derivatives thereof or its combination;
R1 is bidentate containing n-donor ligand, is preferably selected from: DAMBO (boron two pyrroles's methyl-derivatives), o-phenylenediamine analog derivative, diaminourea fluorescein derivative, diaminourea Rhodamine Derivatives or 3-formic acid-o-phenylenediamine or its combination;
M is metal.
4. drug system as claimed in claim 1, is characterized in that, described targeting homing device is selected from lower group: folate molecule, galactose molecule, biotin or its combination.
5. drug system as claimed in claim 1, is characterized in that, described carrier is linked by covalent bond and described exogenous NO gas donor; And/or
Described carrier is linked by covalent bond and described targeting homing device.
6. a method of preparing nano composite material drug system as claimed in claim 1, is characterized in that, described method comprises step:
(1) provide exogenous NO gas donor, targeting homing device and carrier nanoparticle;
(2) described exogenous NO gas donor, described targeting homing device and described carrier nanoparticle are carried out to covalency load, thereby form drug system as claimed in claim 1.
7. preparation method as claimed in claim 6, is characterized in that, described method also comprises the following steps of the described exogenous NO gas donor of preparation before in step (1):
(a1) provide formula A compound,
M(tpy′)Cl 3 (A)
Wherein, M is metal, is preferably selected from lower group: Ru, Mn;
Tpy ' is three tooth containing n-donor ligands;
(a2) in inert atmosphere, with described M (tpy ') Cl 3with R 1and NH 4pF 6reaction, forms [(tpy ') M (R 1) Cl] (PF 6) 3; Wherein, R 1be selected from lower group: DAMBO (boron two pyrroles's methyl-derivatives) or o-phenylenediamine;
(a3) in suitable solvent, by described [(tpy ') M (R 1) Cl] (PF 6) 3react with nitrite, obtain [(tpy ') M (R 1) NO 2] (PF 6);
(a4) by [(tpy ') M (R 1) NO 2] (PF 6) and acid and NH 4pF 6reaction, obtains described exogenous NO gas donor.
8. this drug system preparation method according to claim 6, is characterized by, and described method also comprises in step (1) following steps of preparing nanoparticulate carriers before:
By (4-NH 2)-C 6h 4-PO 3h reacts with nanoparticle, obtains the nanoparticulate carriers of surface amination.
9. a pharmaceutical composition, is characterized in that, comprising: the nano composite material drug system as described in as arbitrary in claim 1-5, and pharmaceutically acceptable carrier.
10. the purposes of the nano composite material drug system as described in as arbitrary in claim 1-5, is characterized in that, for the preparation of medicine, described medicine is targeted drug and has visible ray controlled release NO.
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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104829652A (en) * 2015-03-31 2015-08-12 华东理工大学 Near infrared light-controlled nitrogen monoxide release nanosystem, and preparation method and application thereof
CN107961375A (en) * 2017-10-24 2018-04-27 中国科学院高能物理研究所 A kind of nano metal sulfide material and its preparation method and application
TWI624269B (en) * 2016-07-05 2018-05-21 郭文碩 One-and two-photon photoexcited photodynamic therapy and contrast agent with graphene-based quantum dots
CN109289052A (en) * 2018-12-04 2019-02-01 华东理工大学 Nitric oxide and cis-platinum targeting joint controllable drug delivery Nano medication system and preparation
CN110520462A (en) * 2017-04-18 2019-11-29 基础科学研究院 High-molecular gel and its manufacturing method and article comprising the high-molecular gel
CN110632142A (en) * 2019-08-28 2019-12-31 江南大学 Preparation method and application of electrochemical biosensor based on gold palladium-graphene quantum dot composite material
CN111481737A (en) * 2020-05-07 2020-08-04 华东理工大学 Magnetic temperature-sensitive hydrogel for near-infrared light-controlled release of nitric oxide, and preparation and application thereof
CN113082206A (en) * 2021-04-27 2021-07-09 中国药科大学 Macromolecule nitric oxide donor modified up-conversion nanoparticles, preparation method and application

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102167405A (en) * 2011-01-24 2011-08-31 昆明贵金属研究所 New method for preparing ruthenium nitrosylnitrate solid
CN102429868A (en) * 2011-12-09 2012-05-02 南开大学 Liposome medicinal composition with tumor targeting, in-vivo tracing and treating functions and preparation method thereof
CN103520720A (en) * 2013-10-18 2014-01-22 上海交通大学 Preparation method of folic acid coupled carboxymethyl chitosan nanoparticle serving as photo-releasing NO carrier

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102167405A (en) * 2011-01-24 2011-08-31 昆明贵金属研究所 New method for preparing ruthenium nitrosylnitrate solid
CN102429868A (en) * 2011-12-09 2012-05-02 南开大学 Liposome medicinal composition with tumor targeting, in-vivo tracing and treating functions and preparation method thereof
CN103520720A (en) * 2013-10-18 2014-01-22 上海交通大学 Preparation method of folic acid coupled carboxymethyl chitosan nanoparticle serving as photo-releasing NO carrier

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
王兰等: "抗肿瘤一氧化氮供体药物研究进展", 《中国新药杂志》 *

Cited By (12)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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CN110520462A (en) * 2017-04-18 2019-11-29 基础科学研究院 High-molecular gel and its manufacturing method and article comprising the high-molecular gel
CN110520462B (en) * 2017-04-18 2022-07-26 基础科学研究院 Polymer gel, method for producing same, and article containing same
CN107961375A (en) * 2017-10-24 2018-04-27 中国科学院高能物理研究所 A kind of nano metal sulfide material and its preparation method and application
CN107961375B (en) * 2017-10-24 2020-12-08 中国科学院高能物理研究所 Metal sulfide nano material and preparation method and application thereof
CN109289052A (en) * 2018-12-04 2019-02-01 华东理工大学 Nitric oxide and cis-platinum targeting joint controllable drug delivery Nano medication system and preparation
CN109289052B (en) * 2018-12-04 2022-02-08 华东理工大学 Nitric oxide and cis-platinum targeted combined controllable drug delivery nano-drug system and preparation
CN110632142A (en) * 2019-08-28 2019-12-31 江南大学 Preparation method and application of electrochemical biosensor based on gold palladium-graphene quantum dot composite material
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CN113082206A (en) * 2021-04-27 2021-07-09 中国药科大学 Macromolecule nitric oxide donor modified up-conversion nanoparticles, preparation method and application

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