CN105688222A - Application of chitosan metal complex particles as dosage carrier based on active oxygen responsiveness - Google Patents
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/02—Inorganic compounds
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
Abstract
The invention discloses application of chitosan metal complex particles as a dosage carrier based on active oxygen responsiveness. The weight-average molecular weight of chitosan is 60-250 kDa, the degree of deacetylation is 80-95%, and the metal ions are divalent or trivalent metal ions. According to in-vitro ultraviolet turbidity measurement and nile red in-vitro release test results, it is found that the chitosan metal complex particles obtained through different methods have the good oxidation responsiveness, the oxidation response speeds and in-vitro release efficiency of two particles are remarkably increased on the existence condition of metal ions, and the chitosan metal complex particles are expected to be developed to be the novel drug carrier based on active oxygen intelligent responsiveness.
Description
Technical field
The present invention relates to the new opplication of a kind of material, particularly to chitosan metal composition particles as the application responding to drug carrier based on active oxygen。
Background technology
Oxidative stress can cause activity in vivo oxygen (Reactiveoxygenspecies, ROS) change of content, oxidation response material can spontaneously change macroscopic view or the microscopic property of material according to the change of ROS content, and the result of its change mainly includes swelling, dissolving or specific contraction, the hardening etc. of material。This kind of material being directed to vivo oxidation micro-environmental variation is mainly used in strengthening the targeting of pharmaceutical carrier, present stage research is more is the synthetic material by chemical modification, including organometallic polymer, conjugatd polymers, containing heteroatomic organic polymer。The oxidation responsive materials of synthetic, although it is high with ROS reactivity, but their preparation condition, preservation condition require of a relatively high, cytotoxicity is big, internal hold-up is big, the immunoreactivity of human body is strong, and this series of problems limits the use at field of medicaments of the synthesis oxidation responsive materials to a certain extent。Based on this, present people increasingly pay attention to some natural products with oxidation response characteristic, to avoiding a series of defects of chemical synthetic material。
Chitosan metal complexes (CS-Met) is chitosan or derivatives thereof and a kind of material of metal ion reaction generation。With chitosan or modification of chitosan for substrate, the chitosan-based metal complex materials of complexed metal ion gained has the performance of many excellences, good application prospect (Yin Xueqiong is had in enzyme immobilization agent, separation film, artificial Urea absorbent, the growth inducing agent of nano microcrystalline, chemical catalyst, plant growth regulator and the field such as metal separation and content detection, Sun Zhongliang, Lin Qiang etc. chitosan-based metal complex materials and application present situation [J] thereof. Kunming University of Science and Technology's journal)。
Yang Zifang. the synthesis of Chitosan-phospholipid complex metal complex, sign and antibacterial anti-cancer properties research [D]. Shanghai Normal University, 2010, adopting Chitosan-phospholipid complex and rare earth nitrate and transition metal salt is raw material, use reflux condensation mode method and the sedimentation method, synthesize the Chitosan-phospholipid complex metal complex CTS/RE (RE=La of series of new3+, Ce3 +, Ce4+, Pr3+, Nd3+, Sm3+, Eu3+, Y3+, Er3+, Gd3+；CTS is Chitosan-phospholipid complex), find that coordination compound all has good inhibiting effect to testing with bacterium, belong to broad spectrum antimicrobicide, there is good antibacterial durability and long-lasting, and coordination compound to the antibacterial ability of golden yellow glucose coccus more than to colibacillary antibacterial ability。Additionally, part coordination compound also carries out Anticancer Activity in vitro testing research, experimental test target cell is people's chronic myelocytic leukemia K562 cell。In experiment, the cell after K562 cell and coordination compound effect is carried out morphologic observation research by the aobvious mirror of application inversion light, fluorescence microscope and transmission electron microscope etc. respectively, use mtt assay to carry out cytoactive detection, utilize apoptotic cell rate after flow cytomery coordination compound effect。Anti-tumor experiment result shows that growth and the propagation of K562 tumor cell are had obvious inhibitory action by oligochitosan cerium complexes, property that there were significant differences compared with matched group, it is suppressed that rate improves with the increase of dosage within the specific limits。
Existing research focuses primarily upon antibacterial, the anti-tumor activity of chitosan metal complexes, but yet there are no the report using chitosan metal composition particles as oxidation response medicine carrier。
Summary of the invention
It is an object of the invention to provide chitosan metal composition particles as the application responding to drug carrier based on active oxygen。
The technical solution used in the present invention is:
The chitosan metal composition particles that a kind of active oxygen (hydrogen peroxide) responds, the weight average molecular weight of chitosan is 60kDa～250kDa, and deacetylation is 80～95%, and metal ion is bivalence or trivalent metal ion。
Microgranule is micron order microgranule and nano_scale particle, particularly including particle diameter microgranule between 10nm～1 μm。
As the further improvement of above-mentioned chitosan metal composition particles, metal ion is selected from copper ion, calcium ion, zinc ion, iron ion。
In chitosan metal complexes, the use level of metal ion can be adjusted as required, it is preferable that the use level of metal ion is the 50～100% of its maximum use level, and the best is 50～90%。The limit that the definition of maximum use level can coordinate after chitosan or chitosan particle are mixed with the water soluble salt of enough corresponding metal ions and fully reacted。Especially, the mass ratio that feeds intake of chitosan and slaine is (1:1)～(5:1), more preferably be (1:1)～(2.5:1), more preferably (1.67:1)～(2.5:1)。
The preparation method of above-mentioned chitosan metal composition particles, comprises the steps:
1) being mixed by the water-soluble salt solution of chitosan particle and a metal ion species, prepare chitosan metal composition particles, wherein, the weight average molecular weight of chitosan is 60k～250kDa, and deacetylation is 80～95%(A type carrier)；
Or
A) water-soluble salt solution of chitosan and a metal ion species is mixed, prepare chitosan metal complexes；
B) chitosan metal complexes is mixed with sodium polyphosphate, prepare chitosan metal composition particles (Type B carrier) by ionic cross-linking；
Wherein, the weight average molecular weight of chitosan is 60k～250kDa, and deacetylation is 80～95%。
Further improvement as above-mentioned preparation method, when preparing chitosan metal complexes or chitosan metal composition particles, the mass ratio that feeds intake of chitosan or chitosan particle and slaine is (1:1)～(5:1), it is preferably (1:1)～(2.5:1), more preferably (1.67:1)～(2.5:1), reaction temperature is 20～30 DEG C, and magnetic stirring apparatus rotating speed is 100～200rpm, and the response time is 1～4h。Mass ratio refers to mass ratio during dry weight。
Further improvement as above-mentioned preparation method; step 1) prepares chitosan particle or when step b) prepares chitosan metal composition particles; the mass ratio of chitosan and sodium polyphosphate is 6.25:1; reaction temperature is 20～30 DEG C; magnetic stirring apparatus rotating speed is 500～600rpm; response time is 20min～60min, pH is 5.3～5.5。
Especially, the preparation method of chitosan metal composition particles, comprise the steps:
1) chitosan is dissolved in 1% acetum it is sufficiently stirred for 1h, regulate pH value with the sodium hydroxide solution of 1.0mol/L and stir 0.5h after 5.3～5.5,8.0mL polyphosphoric acids sodium solution (1.0mg/mL) is slowly added dropwise in 20mL chitosan-acetic acid solution (2.5mg/mL) under 500～600rpm rotating speed, after hybrid reaction 0.5h, the centrifugal 10min of 3000rpm takes supernatant, namely obtains chitosan particle after lyophilization；
2) by after chitosan particle solution and metal salt solution hybrid reaction 3h, the centrifugal 10min of 3000rpm takes supernatant, namely obtains chitosan metal composition particles (A type carrier) after lyophilization。
Or comprise the steps:
A) chitosan aqueous solution and reacting metal salt solution 3h, then room temperature stands, and sucking filtration takes precipitation, washs successively 3 times with 3 times of volume distilled water and 3 times of volume dehydrated alcohol, and product vacuum is drying to obtain chitosan metal complexes；
B) chitosan metal complexes is dissolved in 1% acetum and is sufficiently stirred for 1h, then regulate pH value with the sodium hydroxide solution of 1.0mol/L and stir 0.5h after 5.3～5.5, again 8.0mL polyphosphoric acids sodium solution (1.0mg/mL) is slowly added dropwise in 20mL chitosan metal complexes solution (2.5mg/mL) under 500～600rpm rotating speed, after hybrid reaction 0.5h, the centrifugal 10min of 3000rpm takes supernatant, and namely lyophilization obtains chitosan metal composition particles (Type B carrier)。
Chitosan metal composition particles as the application responding to drug carrier based on active oxygen, wherein chitosan metal composition particles as it has been described above, or above-mentioned preparation method prepare。
The invention has the beneficial effects as follows:
According to external ultraviolet turbidimetric analysis turbidimetry, Nile red extracorporeal releasing test result of the test, finding that the chitosan metal composition particles for preparing of distinct methods is respectively provided with and well aoxidize response, under the existence condition of metal ion, the oxidation speed of response of two kinds of microgranules and release in vitro efficiency all increase significantly；And relatively Type B carrier is strong for the oxidation responding ability of A type carrier；The oxidation sensitive of each metallic ion coordination is also different, and the sensitivity after coordinating with copper ion is the strongest。According to cytotoxicity experiment and Experiment of Histocompatibility result, A type carrier and Type B carrier to histoorgan all without obvious inflammatory reaction, it is shown that good histocompatibility。
Chitosan metal composition particles is expected to exploitation for the intelligent response new drug carrier based on active oxygen。
Accompanying drawing explanation
Fig. 1～9 are different chitosan metal composition particles testing results of ultraviolet turbidity under hydrogen peroxide；
Figure 10～12 are the fluorescence intensity testing results after different chitosan metal composition particles bag load Nile red under Hydrogen Peroxide；
Figure 13～21 are the fluorescence intensity testing results after different molecular weight chitosan metal composition particles bag load Nile red under Hydrogen Peroxide；
Figure 22 and 23 are the cytotoxicity experiment results of different chitosan metal composition particles；
Figure 24 and 25 are the Experiment of Histocompatibility results of chitosan-calciumion complexes particles A type carrier and Type B carrier。
Detailed description of the invention
The preparation of chitosan particle
With reference to Fan, W., etal., Formationmechanismofmonodisperse, lowmolecularweightchitosannanoparticlesbyionicgelationte chnique [J] .ColloidsandSurfacesB:Biointerfaces, 2012.90 (21): p.21-27. disclosed method, concretely comprising the following steps of chitosan particle (CSNPs) is prepared: be dissolved in by 50mg chitosan in 1% acetum forming concentration is the chitosan solution of 2.5mg/mL according to Ionic gelation method principle, after magnetic agitation 1h, pH to 5.3-5.5 is adjusted with 1.0mol/L sodium hydroxide solution, continue stirring 0.5h。Being slowly added dropwise under 500～600rpm rotating speed in 20mL chitosan-acetic acid solution by 8.0mL polyphosphoric acids sodium solution (1.0mg/mL), after hybrid reaction 0.5h, take supernatant after centrifugal (3000rpm) 10min, namely lyophilization obtains CSNPs。
Chitosan metal composition particles CSNPs-Met(A type carrier) preparation
The preparation of chitosan particle-copper ion coordination compound (CSNPs-Cu)
5mL copper chloride solution (4.0mg/mL) mixes with 20mL chitosan particle (2.5mg/mL), and 200rpm magnetic agitation is about 3h, takes supernatant after centrifugal (3000rpm) 10min, and namely lyophilization obtains CSNPs-Cu。The molecular weight of the chitosan weight average used is 60kDa, and deacetylation is 95%。
The preparation of chitosan particle-calcium ion complexes (CSNPs-Ca)
5mL calcium chloride solution (6.0mg/mL) mixes with 20mL chitosan particle (2.5mg/mL), and 200rpm magnetic agitation is about 3h, takes supernatant, obtain lyophilization CSNPs-Ca after centrifugal (3000rpm) 10min。The molecular weight of the chitosan weight average used is 100kDa, and deacetylation is 89%。
The preparation of chitosan particle-zinc ion coordination thing (CSNPs-Zn)
5mL solution of zinc sulfate (6.0mg/mL) mixes with 20mL chitosan particle (2.5mg/mL), and 200rpm magnetic agitation is about 3h, takes supernatant, obtain lyophilization CSNPs-Zn after centrifugal (3000rpm) 10min。The molecular weight of the chitosan weight average used is 150kDa, and deacetylation is 80%。
The preparation of chitosan particle-ferric iron complexes (CSNPs-Fe)
5mL ferrum sulfuricum oxydatum solutum (6.0mg/mL) mixes with 20mL chitosan particle (2.5mg/mL), and 200rpm magnetic agitation is about 3h, takes supernatant after centrifugal (3000rpm) 10min, and namely lyophilization obtains CSNPs-Fe。The molecular weight of the chitosan weight average used is 250kDa, and deacetylation is 86%。
Chitosan metal composition particles (CS-Met) NPs(B type carrier) preparation
The preparation of chitosan-copper ion coordination compound (CS-Cu)
5mL copper chloride solution (4.0mg/mL) mixes with 20mL chitosan aqueous suspension (2.5mg/mL), and 200rpm magnetic agitation is about 3h, and reaction temperature is 20 ~ 30 DEG C, standing makes precipitation assemble, vacuum pump sucking filtration, with the distilled water wash 3 times of 3 times of volumes, then with absolute ethanol washing 3 times。By product room temperature vacuum drying after washing, obtain CS-Cu。The molecular weight of the chitosan weight average used is 60kDa, and deacetylation is 95%。
The preparation of chitosan-calcium ion complexes (CS-Ca)
5mL calcium chloride solution (6.0mg/mL) mixes with 20mL chitosan aqueous suspension (2.5mg/mL), and 200rpm magnetic agitation is about 3h, and reaction temperature is 20 ~ 30 DEG C, standing makes precipitation assemble, vacuum pump sucking filtration, with the distilled water wash 3 times of 3 times of volumes, then with absolute ethanol washing 3 times。By product room temperature vacuum drying after washing, obtain CS-Ca。The molecular weight of the chitosan weight average used is 100kDa, and deacetylation is 89%。
The preparation of chitosan-zinc ion coordination thing (CS-Zn)
5mL solution of zinc sulfate (6.0mg/mL) mixes with 20mL chitosan aqueous suspension (2.5mg/mL), and 200rpm magnetic agitation is about 3h, and reaction temperature is 20 ~ 30 DEG C, standing makes precipitation assemble, vacuum pump sucking filtration, with the distilled water wash 3 times of 3 times of volumes, then with absolute ethanol washing 3 times。By product room temperature vacuum drying after washing, obtain CS-Zn。The molecular weight of the chitosan weight average used is 150kDa, and deacetylation is 80%。
The preparation of chitosan-ferric iron complexes (CS-Fe)
5mL ferrum sulfuricum oxydatum solutum (6.0mg/mL) mixes with 20mL chitosan aqueous suspension (2.5mg/mL), and 200rpm magnetic agitation is about 3h, and reaction temperature is 20 ~ 30 DEG C, standing makes precipitation assemble, vacuum pump sucking filtration, with the distilled water wash 3 times of 3 times of volumes, then with absolute ethanol washing 3 times。By product room temperature vacuum drying after washing, obtain CS-Fe。The molecular weight of the chitosan weight average used is 250kDa, and deacetylation is 86%。
The preparation of chitosan-copper ion composition particles ((CS-Cu) NPs)
50mgCS-Cu is dissolved in 1% acetum and forms the CS-Cu acetum that concentration is 2.5mg/mL, after magnetic agitation 1h, adjust pH to 5.3-5.5 with 1.0mol/L sodium hydroxide solution, continue stirring 0.5h。Being slowly added dropwise under 600rpm rotating speed in CS-Cu acetum by 8.0mL polyphosphoric acids sodium solution (1.0mg/mL), after hybrid reaction 0.5h, take supernatant after centrifugal (3000rpm) 10min, namely lyophilization obtains (CS-Cu) NPs。
The preparation of chitosan-calcium ion complexes microgranule ((CS-Ca) NPs)
50mgCS-Ca is dissolved in 1% acetum and forms the CS-Ca acetum that concentration is 2.5mg/mL, after magnetic agitation 1h, adjust pH to 5.3-5.5 with 1.0mol/L sodium hydroxide solution, continue stirring 0.5h。Being slowly added dropwise under 600rpm rotating speed in CS-Ca acetum by 8.0mL polyphosphoric acids sodium solution (1.0mg/mL), after hybrid reaction 0.5h, take supernatant after centrifugal (3000rpm) 10min, namely lyophilization obtains (CS-Ca) NPs。
The preparation of chitosan-zinc ion coordination thing microgranule ((CS-Zn) NPs)
50mgCS-Zn is dissolved in 1% acetum and forms the CS-Zn acetum that concentration is 2.5mg/mL, after magnetic agitation 1h, adjust pH to 5.3-5.5 with 1.0mol/L sodium hydroxide solution, continue stirring 0.5h。Being slowly added dropwise under 600rpm rotating speed in CS-Zn acetum by 8.0mL polyphosphoric acids sodium solution (1.0mg/mL), after hybrid reaction 0.5h, take supernatant after centrifugal (3000rpm) 10min, namely lyophilization obtains (CS-Zn) NPs。
The preparation of chitosan-ferric iron complexes microgranule ((CS-Fe) NPs)
50mgCS-Fe is dissolved in 1% acetum and forms the CS-Fe acetum that concentration is 2.5mg/mL, after magnetic agitation 1h, adjust pH to 5.3-5.5 with 1.0mol/L sodium hydroxide solution, continue stirring 0.5h。Being slowly added dropwise under 600rpm rotating speed in CS-Fe acetum by 8.0mL polyphosphoric acids sodium solution (1.0mg/mL), after hybrid reaction 0.5h, take supernatant after centrifugal (3000rpm) 10min, namely lyophilization obtains (CS-Fe) NPs。
Contrast microgranule
The chitosan using molecular weight to be 10kDa, 50kDa and 300kDa respectively, prepares the CSNPs of different molecular weight, CSNPs-Cu, CSNPs-Ca, CSNPs-Zn, CSNPs-Fe, (CS-Cu) NPs, (CS-Ca) NPs, (CS-Zn) NPs, (CS-Fe) NPs microgranule by the method for above-mentioned correspondence。
Dynamic Light Scattering Determination
Take CSNPs, A type carrier and each 1mL of Type B carrier, after diluting 10 times, take 3mL, use dynamic light scattering detection each sample light intensity to measure particle diameter。
Experimental result is as shown in table 1。As can be seen from the table: A type carrier and Type B carrier particle are uniformly dispersed, and uniform particle sizes is distributed between 200～400nm scope。The microgranule of different metal complex preparation, particle diameter gap is little。
The particle diameter of table 1CSNPs, A type carrier and Type B carrier and PDI
。
Turbidity ultraviolet detection
Respectively by 2mL chitosan particle, A type carrier and Type B carrier and 2mL2000,1000,300,100mMH2O2Mixing, is placed in constant temperature oscillator (37 DEG C, 100rpm)。Not in the same time, observing the oxidative degradation situation (n=3) of CSNPs, CSNPs-Met and (CS-Met) NPs with ultraviolet-uisible spectrophotometer detection。
Experimental result is such as shown in Fig. 1～9, and the molecular weight of the chitosan weight average of use is 100kDa, and deacetylation is 85%。As can be seen from the figure: A type carrier and Type B carrier will be good than chitosan and chitosan particle to the responding ability of hydrogen peroxide, response order is A type carrier > Type B carrier > chitosan particle > chitosan。
The release in vitro of bag load Nile red carrier measures
This effects chitosan particle, A type carrier, Type B carrier release in vitro behavior, concrete operation step is: 2mL is encapsulated the chitosan particle of Nile red (fluorescent dye, aqueous environments can produce fluorescent quenching), A type carrier, Type B carrier respectively with 2mL2000,1000,300,100mMH2O2Mixing, is placed in constant temperature oscillator (37 DEG C, 100rpm)。After different time sections extraction 0.05mL mixed liquor adds 3mL distilled water diluting, use its fluorescence intensity of fluorescent spectrophotometer assay (n=3)。
The bag support method of Nile red is as follows: be dissolved in 1g Tween 80 by 1mg Nile red, again by itself and polyphosphoric acids sodium solution (8mL, 1.0mg/mL) mixing, is subsequently adding in chitosan or chitosan metal complexes, obtains the microgranule of bag load Nile red by the preparation method of blank microparticles。Or prepare by other known methods。
After each carrier bag load Nile red, the change of fluorescence intensity in hydrogen peroxide of each carrier is detected with spectrofluorophotometer, embodiment CSNPs, CSNPs-Cu, CSNPs-Ca, CSNPs-Zn, CSNPs-Fe, (CS-Cu) NPs, (CS-Ca) NPs, (CS-Zn) NPs, (CS-Fe) NPs experimental result such as shown in Figure 10～12, the molecular weight of the chitosan weight average used is 100kDa, and deacetylation is 85%；The experimental result of the contrast microgranule prepared by 10kDa, 50kDa and 300kDa respectively such as Figure 13～15, shown in 16～18,19～21。
Can be seen that from result, A type carrier and Type B carrier are at H2O2Nile red and H all can be discharged under condition2O2Concentration is more high, and rate of release is more fast, and when there is no H2O2When existing, fluorescent probe is without discharging。Show that two kinds of carriers are respectively provided with oxidation response, without H2O2Water in can remain stable for for a long time；And run into H2O2Just can being destroyed thus discharging Nile red, and the more high release of reactive oxygen species is more fast, namely rate of release presents H2O2Concentration dependent。And, add the oxidation sensitive of microgranule after chitosan particle and metal ion complexation, accelerate rate of release。Result shows, A type carrier is best to the oxidation response performance of hydrogen peroxide, and response order is A type carrier > Type B carrier > chitosan particle > chitosan。
The particle metal coordination compound that different molecular weight chitosan prepares is to H2O2There is good response performance, but it can be seen that chitosan molecule amount is within the scope of 60kDa～250kDa, the particle metal coordination compound prepared is to H2O2Response best。
Cytotoxicity experiment
Mouse monokaryon macrophage leukaemia RAW264.7 is with 1 × 106Density cultivate containing 1mL height sugar DEME basal medium, 10% hyclone (FBS) 100U/mL 24 orifice plates in, chitosan, chitosan particle, A type carrier, Type B carrier is added respectively after 24h, concentration respectively 25,50,100,200 and 500 μ g/mL process 2 days, then measure the apoptosis of cell with flow cytometer。
As depicted in figures 22 and 23, the molecular weight of the chitosan weight average of use is 100kDa to experimental result, and deacetylation is 85%。Compared with chitosan, chitosan particle, A type carrier and Type B carrier are relatively low to the toxicity of RAW264.7 cell or do not have toxicity under concentration conditions low, middle。And under a high concentration condition, only CSNPs-Zn has bigger toxicity, all the other A type carriers and Type B carrier toxicity are relatively low or do not have toxicity, illustrate that A type carrier and Type B carrier can as the drug administration carriers of safety。
Experiment of Histocompatibility
Weighing 250mg chitosan, chitosan particle, A type carrier and Type B carrier respectively, add 6mLpH2.0 dissolving with hydrochloric acid, 1.0mol/LNaOH adjusts pH value to 7.4, puts in 10mL measuring bottle, is diluted with water to scale, prepares 25mg/mL solution。Take body weight 18-22g healthy mice, single intraperitoneal injection solution (1000mg/kg)。Putting to death mice after administration 14d, take main organs (heart, liver, lung, kidney, brain) HE dyeing, after section, each internal organs of basis of microscopic observation are with or without inflammatory reaction。
As shown in FIG. 24 and 25, the molecular weight of the chitosan weight average of use is 100kDa to the experimental result of CSNPs-Ca and (CS-Ca) NPs, and deacetylation is 85%。Tissue slice microexamination shows, two kinds of nano materials to histoorgan all without obvious inflammatory reaction, it is shown that good histocompatibility。
Claims (9)
1. a chitosan metal composition particles for active oxygen response, the weight average molecular weight of chitosan is 60k～250kDa, and deacetylation is 80～95%, and metal ion is bivalence or trivalent metal ion。
2. chitosan metal composition particles according to claim 1, it is characterised in that: metal ion is selected from copper ion, calcium ion, zinc ion, iron ion。
3. chitosan metal composition particles according to claim 1, it is characterised in that: the mass ratio that feeds intake of chitosan and slaine is (1:1)～(5:1)。
4. the preparation method of chitosan metal composition particles described in claim 1, comprises the steps:
1) being mixed by the water-soluble salt solution of chitosan particle and a metal ion species, prepare chitosan metal composition particles, wherein, the weight average molecular weight of chitosan is 60k～250kDa, and deacetylation is 80～95%；
Or
A) water-soluble salt solution of chitosan and a metal ion species is mixed, prepare chitosan metal complexes；
B) chitosan metal complexes is mixed with sodium polyphosphate, prepare chitosan metal composition particles by ionic cross-linking；
Wherein, the weight average molecular weight of chitosan is 60k～250kDa, and deacetylation is 80～95%。
5. preparation method according to claim 4, it is characterized in that: when preparing chitosan metal complexes or chitosan metal composition particles, the mass ratio that feeds intake of chitosan or chitosan particle and slaine is (1:1)～(5:1), reaction temperature is 20～30 DEG C, magnetic stirring apparatus rotating speed is 100～200rpm, and the response time is 1～4h。
6. preparation method according to claim 4; it is characterized in that: step 1) prepares chitosan particle or when step b) prepares chitosan metal composition particles; the mass ratio of chitosan and sodium polyphosphate is 6.25:1; reaction temperature is 20～30 DEG C; magnetic stirring apparatus rotating speed is 500～600rpm; response time is 20min～60min, pH is 5.3～5.5。
7. preparation method according to claim 4, it is characterised in that: comprise the steps:
1) chitosan is dissolved in 1% acetum it is sufficiently stirred for 1h, regulate pH value with the sodium hydroxide solution of 1.0mol/L and stir 0.5h after 5.3～5.5, again 8.0mL polyphosphoric acids sodium solution (1.0mg/mL) is slowly added dropwise in 20mL chitosan-acetic acid solution (2.5mg/mL) under 500～600rpm rotating speed, after hybrid reaction 0.5h, the centrifugal 10min of 3000rpm takes supernatant, namely obtains chitosan particle after lyophilization；
2) by after chitosan particle solution and metal salt solution hybrid reaction 3h, the centrifugal 10min of 3000rpm takes supernatant, namely obtains chitosan metal composition particles after lyophilization。
8. preparation method according to claim 4, it is characterised in that: comprise the steps:
A) chitosan aqueous solution and reacting metal salt solution 3h, then room temperature stands, and sucking filtration takes precipitation, washs successively 3 times with 3 times of volume distilled water and 3 times of volume dehydrated alcohol, and product vacuum is drying to obtain chitosan metal complexes；
B) chitosan metal complexes is dissolved in 1% acetum and is sufficiently stirred for 1h, then regulate pH value with the sodium hydroxide solution of 1.0mol/L and stir 0.5h after 5.3～5.5, again 8.0mL polyphosphoric acids sodium solution (1.0mg/mL) is slowly added dropwise in 20mL chitosan metal complexes solution (2.5mg/mL) under 500～600rpm rotating speed, after hybrid reaction 0.5h, the centrifugal 10min of 3000rpm takes supernatant, and namely lyophilization obtains chitosan metal composition particles。
9. chitosan metal composition particles is as the application responding to drug carrier based on active oxygen, wherein described in chitosan metal composition particles such as claims 1 to 3 any one, or prepares by the preparation method described in claim 4～8 any one。
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006044785A1 (en) * | 2004-10-18 | 2006-04-27 | E.I. Dupont De Nemours And Company | Process for making antimicrobial polymer articles |
| CN101104651A (en) * | 2007-06-22 | 2008-01-16 | 中南大学 | Preparation for modified chitosan metal complexes micro-sphere used for carrying medicament and preparation for the modified chitosan |
-
2016
- 2016-03-14 CN CN201610143602.1A patent/CN105688222A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2006044785A1 (en) * | 2004-10-18 | 2006-04-27 | E.I. Dupont De Nemours And Company | Process for making antimicrobial polymer articles |
| CN101104651A (en) * | 2007-06-22 | 2008-01-16 | 中南大学 | Preparation for modified chitosan metal complexes micro-sphere used for carrying medicament and preparation for the modified chitosan |
Non-Patent Citations (1)
| Title |
|---|
| 张世倡: "基于活性氧响应壳聚糖纳米给药载体的研究", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107991374A (en) * | 2017-11-30 | 2018-05-04 | 江苏省农业科学院 | The method that actin in meat is detected based on CS-Zn NPs sensors |
| CN107991374B (en) * | 2017-11-30 | 2019-12-10 | 江苏省农业科学院 | Method for detecting actin in meat based on CS-Zn NPs sensor |
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