CN106806905A - It is a kind of to collect the fluorescence imaging rare earth upconversion nano pharmaceutical carrier integrated with medicine is carried and its application - Google Patents
It is a kind of to collect the fluorescence imaging rare earth upconversion nano pharmaceutical carrier integrated with medicine is carried and its application Download PDFInfo
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- CN106806905A CN106806905A CN201710001962.2A CN201710001962A CN106806905A CN 106806905 A CN106806905 A CN 106806905A CN 201710001962 A CN201710001962 A CN 201710001962A CN 106806905 A CN106806905 A CN 106806905A
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/001—Preparation for luminescence or biological staining
- A61K49/0013—Luminescence
- A61K49/0017—Fluorescence in vivo
- A61K49/005—Fluorescence in vivo characterised by the carrier molecule carrying the fluorescent agent
- A61K49/0052—Small organic molecules
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5005—Wall or coating material
- A61K9/5021—Organic macromolecular compounds
- A61K9/5031—Organic macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poly(lactide-co-glycolide)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
- A61K9/50—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals
- A61K9/5073—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals having two or more different coatings optionally including drug-containing subcoatings
- A61K9/5078—Microcapsules having a gas, liquid or semi-solid filling; Solid microparticles or pellets surrounded by a distinct coating layer, e.g. coated microspheres, coated drug crystals having two or more different coatings optionally including drug-containing subcoatings with drug-free core
Abstract
Collect the fluorescence imaging rare earth upconversion nano pharmaceutical carrier integrated with medicine is carried and its application the invention discloses a kind of.The nano-medicament carrier includes the rare earth upconversion nano particle (UCNPs) with fluorescence imaging function, the antineoplastic for being supported on up-conversion nanoparticles surface and amphipathic nature polyalcohol molecule DSPE polyethylene glycol (DSPE PEG) for being coated on nanoparticle surface.Nano-medicament carrier size uniformity of the invention, pattern rule, with good water solubility and biocompatibility, surface texture is conducive to making it be enriched in tumor tissues by EPR effects, realizes medicine passive target.
Description
Technical field
The present invention relates to a kind of nano-medicament carrier and its application, and in particular to one kind can simultaneously carry out fluorescence imaging and bag
The rare earth upconversion nano pharmaceutical carrier of carrying anti-tumor medicine and application, belong to nano biological medical domain.
Background technology
Malignant tumour is one of principal disease of influence human health, and at present, chemotherapy, operation, radiotherapy are treating cancers
Three big means.Chemotherapy refers specifically to use chemotherapy malignant tumour, into human body after be distributed to whole body rapidly, be a kind of whole body
Treatment.Because traditional chemotherapeutics does not have obvious targeting, therefore while killing tumour cell, can also damage human body
Normal cell, tissue, organ, produce obvious toxic and side effect, make human body immunity degradation.Meanwhile, traditional chemotherapeutics is also
There are problems that metabolism is fast, be also easy to produce.
With the fast development of nanometer technology, nano-medicament carrier is because size is small, toxicity is low, specific surface area is big, physics
Good chemical stability gradually attracts attention in terms of tumor diagnosis and therapy.Nano-medicament carrier can extend medicine half
Decline the phase, increase blood circulation time, improve absorptivity, increase specificity and targeting, using the high-permeability of solid tumor and stagnant
Staying effect (EPR effects) makes drug-rich in tumor locus and realizes medicine controlled release, is favorably improved drug effect, reduces poison secondary
Effect.Be currently used as the material of nano-medicament carrier mainly have liposome, dendroid nano material, polymer nano material,
Macromolecule micelle, CNT, mesoporous silicon oxide etc., but these pharmaceutical carriers are only used for oncotherapy, without examining
Disconnected function, only forms multiple by fluorescent marker or contrast agent modification using modes such as covalent bond or physical absorptions to its surface
Mould assembly nano material, can just be implemented as the Clinics and Practices integration as mediation, be accurately positioned tumour, implement efficiently to control early
Treat.
The rare earth upconversion nano material (UCNPs) for developing in recent years is due to bio-toxicity is low, penetration depth big, light
Damage the advantages of small, fluorescence background is low, signal to noise ratio is high, be the biological labled material with applications well prospect, i.e., it is a kind of new
Fluorescence probe, the inherent defect such as the easy photobleaching of conventional fluorescent dyestuff, the interference of unstable, background fluorescence can be overcome big, and carefully
Cellular toxicity larger quantum point is compared, and up-conversion nanoparticles bio-toxicity is lower.Meanwhile, up-conversion nano material can also be wrapped
Carrying anti-tumor medicine is used for pharmaceutical carrier, and the mode that contains mainly to form hydrophobic pocket structure, and to contain medicine, surface modification mesoporous
Silica contains medicine, the hollow pore passage structure of formation and contains medicine etc..Document (Kang, X.；Cheng,Z.；Li,C.；Yang,
D.；Shang,M.；Ma,P.；Li,G.；Liu,N.；Lin,J.Core–shell structured up-conversion
luminescent and mesoporous NaYF4:Yb3+/Er3+@nSiO2@mSiO2nanospheres as carriers
For drug delivery.J.Phys.Chem.C 2011,115,15801-15811.) prepared by two step sol-gel processes
Go out the UCNPs of mesoporous silicon oxide cladding, antiphlogistic brufen is contained in meso-hole structure, carry out drug encapsulation and release.
Document (Zhang, F.；Braun,G.B.；Pallaoro,A.；Zhang,Y.；Shi,Y.；Cui,D.；Moskovits,M.；
Zhao,D.；Stucky,G.D.Mesoporous multifunctional upconversion luminescent and
magnetic"nanorattle"materials for targeted chemotherapy.Nano Lett.2012,12,61-
67.) a kind of nanometer of core shell structure of ring-type is prepared for by the etching and ion exchange process of surface protection, outside is mesoporous knot
Structure, centre is hollow structure, and inside is solid stratum nucleare, and medicine can be by bag limit in hollow structure.Usual surface mesoporous structure
Stationarity to medicine is poor, medicine easily from hole leakage, it is necessary to surface modify again one layer of polymeric protective layer (Lai,
J.P.；Shah,B.P.；Zhang,Y.X.；Yang,L.T.；Lee,K.B.Real-time monitoring of ATP-
responsive drug release using mesoporous-silica-coated multicolor
upconversion nanoparticles.ACS Nano 2015,9,5234-5245.)。
But above-mentioned medicine-carrying method is required for multistep reaction and strict condition to control, and operating process is relatively cumbersome and time-consuming.
Additionally, up-conversion nanoparticles increased the probability of quenching by repeatedly treatment, up-conversion luminescence efficiency is caused to reduce.For reality
Existing nano-carrier is widely applied in biomedical sector, it is desirable to is prepared by simple synthetic method and is based on upper conversion nano
The nano-medicament carrier of material, and diagnosing tumor is applied to treatment.
The content of the invention
In order to overcome the shortcoming of above-mentioned prior art, a kind of collection fluorescence imaging is carried with the rare earth upconversion nano medicine for carrying medicine
Body, mainly includes：Rare earth upconversion nano particle (UCNPs) with fluorescence imaging function, is supported on up-conversion nanoparticles
The antineoplastic on surface and it is coated on the amphipathic nature polyalcohol molecule distearyl acyl group phosphatidyl ethanol of nanoparticle surface
Amine-polyethylene glycol (DSPE-PEG).
Preferably, the chemical composition of the rare earth upconversion nano particle is NaYF4:Yb, Ln, Ln are Er or Tm, more
Preferably crystallinity is high, the uniform hexagonal phase of pattern is nanocrystalline.
Preferably, the antineoplastic is the prodrug for having modified long chain alkyl group, wherein prodrug is selected from 7-
Ethyl -10-Hydroxycamptothecin (SN38) and Cabazitaxel medicine (CTX).Simple prodrug can not be hydrophobic with UCNPs
Layer is assembled, may be relevant with the rigid planar structure of its own, and after having modified long-chain, can freely be revolved with high resiliency
Turn, the hydrophobic effect between enhancing and oleic acid molecular realizes assembling.On the other hand, these prodrugs can be dissolved in the water.
Wherein, the long chain alkyl group is preferably chain alkyl aliphatic acid, and the aliphatic acid may be selected saturated fatty acid or not
Saturated fatty acid, preferably unrighted acid, more preferably oleic acid, linoleic acid or leukotrienes, most preferably linoleic acid or Asia
Numb acid.
The pro-drug structural formula is one of (I-1) to (I-2) described compound：
The molecular weight of the amphipathic nature polyalcohol molecule DSPE-PEG (DSPE-PEG)
There are 750,2000,5000,10000 etc., preferably molecular weight 2000, i.e. DSPE-PEG2000。
Select amphipathic molecule DSPE-PEG to be coated, mainly have the advantage that：(1) DSPE-PEG is PEGylation
Phospholipid molecule, with good water solubility and biocompatibility, is ratified for human body by FDA；(2) aliphatic chain of DSPE-PEG
Unrighted acid structure with the oleic acid molecular on UCNPs surfaces, antineoplastic is very much like, it is easy to sent out by hydrophobic effect
Raw Supramolecular Assembling, is adsorbed；(3) one layer of hydrophilic layer is formed after nanoparticle surface modified DSPE-PEG, it is to avoid hydrophobic to receive
Rice corpuscles surface and reticuloendothelial system (RES) cell, by RES cellular uptakes, removing, are entered by non-specific hydrophobic effect
And extend the half-life period in blood circulation and improve stability, be conducive to realizing medicine passive target by EPR effects.
On the other hand, the fluorescence imaging rare earth upconversion nano medicine load integrated with medicine is carried is collected the invention provides a kind of
The simple method for preparing of body, the preparation method operating procedure is simple and easy to apply, low cost.
Methods described includes：
(1) it is raw material using rare earth-iron-boron, oleic acid is part, oleic acid/1- octadecylenes are mixed solvent, molten using high temperature
The rare earth upconversion nano particle (OA-UCNPs) of the hot method synthesis oleic acid parcel of agent；
Rare earth-iron-boron is preferably YCl3、YbCl3And ErCl3/TmCl3Or its hydrate.
(2) by the up-conversion nanoparticles, DSPE-PEG2000, anti-tumor prodrug be dispersed in same medium, make
Three is sufficiently mixed with stir in situ method, hydrophobic interaction occurs each other；
Specifically, first three is dispersed in DMSO solvents respectively, is prepared into mother liquor, concentration is respectively 12mg/mL,
100mg/mL, 20mg/mL.Mix according to a certain percentage, three's mass ratio OA-UCNPs：DSPE-PEG2000：Anti-tumor predrug is
10：(10~20)：(1~10), more preferably 10：20：1.Reaction 4h is stirred at room temperature after mixing.
(3) mixture is added drop-wise in the aqueous solution under ultrasound by nanoprecipitation method, when water phase is diffused into
Synthesis drug-loading nanoparticles；
(4) above-mentioned medicine-carried nano particles are isolated and purified, dries or be dispersed in ultra-pure water.
Preferably, selection supercentrifugal process carries out separating-purifying, rotating speed 15000g, time 10min.Purification procedures
For removing the DSPE-PEG not contained with UCNPs effects2000And antineoplastic.
Present invention also offers a kind of fluorescence imaging that collects anticancer is being prepared with the rare earth upconversion nano pharmaceutical carrier of load medicine
Application in terms of medicine and fluorescence imaging.Nano-medicament carrier of the invention can effectively contain medicine, and glimmering by upper conversion
Photoimaging carries out diagnosing tumor, and medicament slow release and administration are carried out in tumor locus using passive target, while realizing to tumour
Up-conversion fluorescence imaging diagnosis and chemotherapeutic treatment, solve the problems, such as single nano-carrier multifunction.Wherein described tumour includes
Breast cancer, intestinal cancer, lung cancer, stomach cancer or cervical carcinoma.
Preferably, fluorescence imaging selects the 980nm optical fiber coupling diode laser of external power adjustable as sharp
Light emitting source.
Beneficial effects of the present invention：The rare earth upconversion nano medicine of surface PEG can be obtained by single step synthetic method
Carrier, synthesis technique is simple, with low cost, efficiency high, be suitable to produce in enormous quantities；Rare earth upconversion nano medicine of the invention is carried
Body has good dispersiveness, good stability in water；Fluorescence probe and antineoplastic are realized that integration is loaded by the present invention,
Medicament slow release and target administration are carried out under fluorescence imaging guidance, reduces toxic and side effect, improve the accuracy for the treatment of.
Brief description of the drawings
Fig. 1 is from left to right respectively OA-UCNPs+DSPE-PEG in embodiment 22000+ SN38 pro-drugs 1, OA-UCNPs
+DSPE-PEG2000Photo of the DMSO solution shake well of+SN38, SN38 after being added dropwise in water under ultrasound.
Fig. 2 is rare earth upconversion nano pharmaceutical carrier (1 pUCNPs) (grey filled lines) and not pastille prepared by embodiment 2
The nano particle (pUCNPs) (black dotted lines) and DSPE-PEG of thing2000Contain SN38 pro-drugs 1 (PEGylated 1)
The ultraviolet-ray visible absorbing curve of (solid black lines).
Fig. 3 is the graph of a relation of UCNPs carrying drug ratios and different UCNPs/ drug responses mass ratioes, wherein medicine is in (a)
SN38 pro-drugs 1, medicine is CTX pro-drugs 2 in (b).
Fig. 4 for nano-particle transmission electron microscope (TEM) photo (on) and high-resolution-ration transmission electric-lens (HRTEM) photo (under),
(a)：NaYF prepared by embodiment 14:The upper conversion nano particle (OA-UCNPs) of Yb, Er；(b)：DSPE-PEG2000Cladding
NaYF4:The upper conversion nano particle (pUCNPs) of Yb, Er；(c)：Rare earth upconversion nano pharmaceutical carrier (1@prepared by embodiment 2
pUCNPs).Black and white engineer's scale represents 100nm and 5nm respectively.
Fig. 5 is to turn on the rare earth of rare earth upconversion nano particle (OA-UCNPs) prepared by embodiment 1 and the preparation of embodiment 2
Change nano-medicament carrier (1 pUCNPs) and not the dynamic light scattering grain size distribution of the nano particle (pUCNPs) of drug containing
(a) and zeta potential diagrams (b), in figure, dhParticle dynamic light scattering particle diameter in a solvent is represented, PDI is polydispersity index.
Fig. 6 is to turn on the rare earth of rare earth upconversion nano particle (OA-UCNPs) prepared by embodiment 1 and the preparation of embodiment 2
Change nano-medicament carrier (1 pUCNPs) and not the nano particle (pUCNPs) of drug containing under 980nm light source activations upper turn
Change fluorescence spectrum to compare, wherein the concentration of UCNPs is 1.5mg/mL, OA-UCNPs is dispersed in hexamethylene, afterwards both dispersions
In water.
Fig. 7 is the rare earth upconversion nano pharmaceutical carrier (1@pUCNPs) of the preparation of embodiment 2 in 37 DEG C, 0.2%Tween
Drug accumulation release profiles in 80 dissolution mediums.
Fig. 8 is that rare earth upconversion nano pharmaceutical carrier (1@pUCNPs) prepared by embodiment 2 and Bcap-37 cell incubations are common
Co-focusing imaging photo after culture 4h.A () Green fluorescence and red fluorescence channel are the fluorescence of UCNPs；In (b)
UCNPs channel selectings be red band up-conversion fluorescence.
Fig. 9 is that rare earth upconversion nano pharmaceutical carrier (1@pUCNPs) prepared by embodiment 2 is anti-swollen in live body level
Knurl effect assessment, (a)：Breast cancer Bcap37 cells tumor bearing nude mice is through the tumor volume change curve after different pharmaceutical treatment；
(b)：Tumor bearing nude mice processed through different pharmaceutical after the tumor quality of the 24th day.
Figure 10 is that different pharmaceutical is processed on human mammary cancer line Bcap-37 tumor bearing nude mice changes of weight influence figure.
In figure, linolenic SN38 has been modified in 1 expression, and linolenic kappa has been modified in 2 expressions
He matches, and Saline represents physiological saline, and SN38 represents SN38, and CPT-11 represents Irinotecan.
Specific embodiment
The present invention is described in further detail with reference to the accompanying drawings and detailed description, but the invention is not restricted to this.
The synthesis of the rare earth upconversion nano particle (UCNPs) of embodiment 1
Accurately weigh six chloride hydrate yttrium (YCl3·6H2O) (0.2366g, 0.78mmol), six hydrous ytterbium chlorides
(YbCl3·6H2O) (0.0775g, 0.20mmol) and anhydrous erbium chloride (ErCl3) (0.0055g, 0.02mmol) addition is extremely
In 100mL round-bottomed flasks, 6mL OA and 15mL 1-ODE are added, be sufficiently mixed stirring.N2100 DEG C are warming up under protection except O2
And H2O, is continuously heating to 160 DEG C of reaction 40min and is completely dissolved to reactant, obtains light yellow clear settled solution, is cooled to
Room temperature.Separately weigh NH4F 0.1482g (4mmol) and NaOH 0.1g (2.5mmol) are dissolved in 10mL methyl alcohol, ultrasonic disperse, acute
Before being added dropwise under strong stirring in cooling solution.Be warming up to 50 DEG C, stir 1h, then heat to 100 DEG C, except methyl alcohol and
Water.After methyl alcohol volatilization is complete, N2300 DEG C of reaction 1.5h are heated under protection.Room temperature is cooled to, a large amount of ethanol, 4320g are added
Centrifugation 8min.After precipitation is disperseed with hexamethylene, plus ethanol centrifugation, so with hexamethylene and ethanol cyclic washing three times, vacuumize
Dry.
By the appropriate rare earth upconversion nano particle NaYF for preparing4:Yb, Er are dispersed in cyclohexane solution, spherical dropwise addition
Onto copper mesh, observed under transmission electron microscope (TEM) and high-resolution-ration transmission electric-lens (HRTEM) respectively.From Fig. 4 (a), preparation
Up-conversion nanoparticles size uniformity, particle diameter is 44.0 ± 1.7nm.Clearly lattice fringe is can see in HRTEM figures, is shown
Nano-particle has good crystallinity, the crystal face correspondence hexagonal structure NaYF of the spacing 0.518nm indicated in figure4:Yb, Er's
(100) crystal face.
The nanoprecipitation method of embodiment 2 prepares rare earth upconversion nano pharmaceutical carrier
By up-conversion nanoparticles (OA-UCNPs), (DSPE- of DSPE-PEG 2000
PEG2000), leukotrienes modification SN38 pro-drugs 1 distinguish ultrasonic disperse in DMSO solution, concentration be respectively 12mg/mL,
100mg/mL、20mg/mL.Three kinds of dispersion liquids are sufficiently mixed, at room temperature slowly vibrating 4h, are then added dropwise over surpassing under ultrasound
In pure water, continue to shake 30min.Centrifugation (15000g, 10min), abandoning supernatant.After the scrubbed centrifugation twice of precipitation, freezing
It is dried overnight.As control, by OA-UCNPs, DSPE-PEG2000, simple SN38 molecules DMSO solution mixing vibration, ultrasound
Under be added drop-wise in ultra-pure water.Prepared to be changed on the rare earth of the Cabazitaxel pro-drug 2 that leukotrienes is modified with same method and received
Meter Zai Ti.The nano-carrier for loading two kinds of medicines is expressed as 1@pUCNPs and 2@pUCNPs.
As shown in figure 1, OA-UCNPs+DSPE-PEG2000+ 1 is mixed with the colloidal solution that Nano medication is clarified, and
OA-UCNPs+DSPE-PEG2000There is white opacity in instilling water in+SN38 mixing, and phenomenon is consistent in instilling water with simple SN38,
Show SN38 unentrappeds, illustrate that the prodrug for not modifying chain alkyl merely can not be assembled with the hydrophobic layer of UCNPs,
May be relevant with the rigid planar structure of its own, and after having modified long-chain, be freely rotatable with high resiliency, strengthen and oleic acid
Hydrophobic effect between molecule realizes assembling.
The uv-visible absorption spectra of upper conversion nano pharmaceutical carrier of prodrug 1 is contained as shown in Fig. 2 having at 367nm
Obvious SN38 characteristic absorption peaks, with only DSPE-PEG2000The nano-particle (PEGylated 1) that cladding prodrug 1 is formed
Absworption peak is substantially overlapping, shows that UCNPs will not be impacted to SN38 absworption peaks, can be with this characteristic absorption peak come quantitative 1@
Active SN38 contents in pUCNPs.
Conversion nano pharmaceutical carrier drug encapsulation rate is determined on the middle rare earth of 3 embodiment of embodiment 2
Fixed OA-UCNPs and DSPE-PEG2000Amount (mass ratio 1:2) prodrug 1 of different quality, is added, is carried out fully
After containing, the amount of the medicine 1 being loaded on UCNPs is determined by ultraviolet-visible absorption spectroscopy (367nm absworption peaks), calculate and carry medicine
Rate.The drugloading rate of prodrug 2 is determined with similar method.Because the molar absorption coefficient of Cabazitaxel is smaller, UCNPs absworption peak meetings
Influence it to determine, therefore collect centrifuged supernatant and washing lotion, indirect determination drug encapsulation is carried out in determine unentrapped 2, wherein purple
Outer characteristic absorption peak is 274nm, while deducting DSPE-PEG at phase co-wavelength2000Absorption.Parallel determination three times.
Fig. 2 is drug encapsulation rate measurement result, when the mass ratio that feeds intake of medicine/UCNPs is 1, reaches SN38 prodrugs 1
Saturation carrying drug ratio, about 12.3%.And the carrying drug ratio of Cabazitaxel prodrug 2 is 18.9%.In order to ensure prodrug can be contained completely
As in UCNPs, the mass ratio that feeds intake of the OA-UCNP/DSPE-PEG2000/1 used in following examples is fixed as 10:20:1.
The sign of conversion nano pharmaceutical carrier on the middle rare earth of 4 embodiment of embodiment 2
The pattern of nano-particle is characterized respectively using transmission electron microscope (TEM) and high-resolution-ration transmission electric-lens (HRTEM), during sample preparation
Nano-particle is dispersed in H2In O, drop is added drop-wise on copper mesh with spherical, electric Microscopic observation again after drying, within sweep of the eye
Randomly selecting 200 particles carries out size statistical analysis.Dynamic light scattering (DLS) particle diameter and zeta current potentials of particle use Ma Er
Literary Particle Size Analyzer is determined.Fluorescence spectrum characterization is characterized using the XRF of external 980nm solid state lasers.
PUCNPs, 1@pUCNPs TEM and HRTEM characterization results it is as shown in Figure 4.Particle diameter be respectively 49.5 ± 1.5nm and
50.3 ± 1.5nm, compares with OA-UCNPs, and particle diameter increased about 5nm, shows successfully to have contained DSPE-PEG2000.DLS particle diameters
Analysis result and zeta current potential characterization results are as shown in Figure 5.Kinetic diameters of the OA-UCNPs in hexamethylene is 91.94nm,
The kinetic diameter of pUCNPs and 1@pUCNPs in water is respectively 113.0nm and 119.7nm, DLS peak shape are sharp, shows do not have
Obvious reunion, increased particle diameter and DSPE-PEG2000Hydrated sheath size be consistent.The zeta electricity of pUCNPs and 1@pUCNPs
Position is respectively -6.28mV and -5.79mV, due to DSPE-PEG2000In there is PO4 -, current potential is somewhat born partially, but numerical value is small, surface
Electric charge influences smaller to carrier transportation.
Fig. 6 compares rare earth upconversion nano particle prepared by above-mentioned nano-medicament carrier (1@pUCNPs) and embodiment 1
(OA-UCNPs) and not up-conversion fluorescence spectrum of the nano particle (pUCNPs) of drug containing under 980nm light source activations, phase
Compared with the fluorescence spectrum of same concentrations OA-UCNPs hexamethylene dispersion liquids, the weak half of fluorescence spectrums of the 1@pUCNPs in water,
Because water has absorption to the exciting light of 980nm, the high energy vibration of hydrone can strengthen Er in addition3+Nonradiative relaxation process
So as to cause fluorescent quenching, but the fluorescence intensity of above-mentioned nano-medicament carrier remains able to meet the demand of imaging.
Conversion nano pharmaceutical carrier drug release rate is determined on the middle rare earth of 5 embodiment of embodiment 2
The nano-carrier of the prodrug containing 0.2mg/mL 1 prepared in embodiment 2 is fitted into (molecular cut off in bag filter
3500Da), it is put into after sealing in the Tween 80 solution of certain volume 0.2%, is placed in constant-temperature table, is slowly shaken at 37 DEG C.Pre-
At the time point for first setting, the release liquid outside 1mL bag filters is taken out, and supplement 0.2% Tween 80 solution of equivalent.Using ultraviolet
The characteristic absorption that absorption process determines 367nm carrys out the active SN38 for quantitatively discharging, and calculates release efficiency, parallel determination three times.
, at 37 DEG C, 0.2%Tween 80 discharges rare earth upconversion nano pharmaceutical carrier (1@pUCNPs) prepared by Fig. 7 displays
Drug accumulation releasing result in medium, curve shows that drug release rate is slower, release efficiency is added up during 96h and is about 36%,
Sustained release is conducive to keep drug effect.
Conversion nano pharmaceutical carrier vitro Cytotoxicity Evaluation on the middle rare earth of 6 embodiment of embodiment 2
Cell in exponential phase is inoculated in (5000 cells/wells), 37 DEG C, 5%CO in 96 orifice plates2, saturation
24h is cultivated in the constant incubator of humidity, nutrient solution is abandoned in suction, culture mediums of the 100 μ L containing the following material of various concentrations is added per hole,
Respectively pUCNPs, CPT-11, the@pUCNPs of SN38, PEGylated 1 and 1, using CPT-11, SN38, PEGylated 1 as
Control, every kind of medicine each concentration 4 multiple holes of setting continue to cultivate 48 or 72h after adding medicine in cell culture incubator.Discard culture
Liquid, 100 μ L CCK-8 solution (mother liquor fresh culture dilutes 10 times) are added per hole, continue to cultivate 30min to 2h, use enzyme mark
The absorbance in each hole, calculates cell survival rate at instrument detection 450nm, draws cell survival curve, obtains medicine cell growth
IC50(half-inhibition concentration).In vitro toxicity knots of the@pUCNPs of rare earth upconversion nano pharmaceutical carrier 1 to various tumour cells
Fruit is shown in Table 1.
In vitro toxicity evaluation result (μM) of each trial drug of table 1
From table 1, after the@pUCNPs of rare earth upconversion nano pharmaceutical carrier 1 and cell incubation 48h, to breast cancer cell
It is the IC of Bcap-3750It is 2.24 ± 0.25 μM to be worth, and its antitumor activity is 26 times of CPT-11；To cervical cancer tumer line HeLa
IC50It is 5.85 ± 1.96 μM to be worth, and its antitumor activity is 9 times of CPT-11；To the IC of lung cancer cell line A54950It is worth and is
0.76 ± 0.09 μM, its antitumor activity is 45 times of CPT-11；To the IC of Gastric caicinoma cell line SGC-790150Be worth for 10.81 ±
2.83 μM, its antitumor activity is 16 times of CPT-11.After 1@pUCNPs and cell incubation 72h, to four kinds of survival rates of cell
Influence becomes apparent from.Wherein to the IC of breast cancer cell line Bcap-3750It is 0.23 ± 0.05 μM to be worth, and its antitumor activity is CPT-
122 times of 11；To the IC of cervical cancer tumer line HeLa50It is 1.69 ± 0.34 μM to be worth, and its antitumor activity is 18 times of CPT-11；
To the IC of lung cancer cell line A54950It is 0.16 ± 0.03 μM to be worth, and its antitumor activity is 52 times of CPT-11；To gastric carcinoma cell lines
The IC of SGC-790150It is 1.75 ± 0.33 μM to be worth, and its antitumor activity is 22 times of CPT-11.
Cytotoxicity result shows that the@pUCNPs of rare earth upconversion nano pharmaceutical carrier 1 have lure more more obvious than CPT-11
The ability of apoptosis of tumor cells is led, the nanoparticle being assembled into simple SN38 and SN38-PEG has similar antitumous effect,
Show that 1@pUCNPs can have effectively to be absorbed by cell, and discharge active SN38 molecules.
The imaging performance of conversion nano pharmaceutical carrier and cellular uptake behavior on the middle rare earth of 7 embodiment of embodiment 2
To evaluate upper conversion nano pharmaceutical carrier imaging performance on a cellular level and investigating cell to above-mentioned carrier
Intake behavior, carry out observational study using common focus point migration fluorescence microscope (CLSM).Bcap-37 after will be adherent and
Culture medium, FITC-dextran compounds (final concentration 0.1mg/mL) containing 1@pUCNPs (the μ g/mL of UCNPs EPCs 100)
4h is cultivated together.Remove the UCNPs and FITC-dextran of excess for 3 times with PBS, consolidated with the PBS solution of 4% paraformaldehyde
Determine 20min, then fully washed with PBS, nucleus dyes 15min with Hoechst 33258 (20 μ g/mL).It is abundant with PBS again
Washing, adds 1.5mL PBS to observe fluorescence imaging under CLSM.Wavelength parameter is selected：Hoechst：405nm wavelength is excited,
425-475nm wave-length coverages are collected；FITC-dextran：488nm is excited, 500-600nm scale collections；UCNPs：980nm swashs
Hair, collects green fluorescence and red fluorescence in 495-550nm and 575-650nm respectively.
The nano-medicament carrier of preparation and the co-focusing imaging result of Bcap-37 cell incubations are as shown in Figure 8.In Fig. 8 (a)
The green and red up-conversion fluorescence become clear in cell are shown, both are superposed to yellow, show that 1@UCNPs can be had by cell
Effect intake.In order to further study endocytosis mode, cultivated together using FITC- glucans, lysosome intake FITC- glucans,
So as to be fluorescently labeled, (green is small for the fluorescence of the fluorescence (dots in red) of 1@pUCNPs of display and FITC- glucans in Fig. 8 (b)
Point) partly overlap, show 1@pUCNPs by lysosome encytosis by cellular uptake, and still have some it is underlapped then because
Medicine is discharged after endocytosis from lysosome.
Result shows that above-mentioned upper conversion nano pharmaceutical carrier can effectively by cell endocytosis, while can be by changing thereon
Fluirescence observation endocytic processes.
Anti-tumor capacity in conversion nano pharmaceutical carrier body on the middle rare earth of 8 embodiment of embodiment 2
In the mammary gland position orthotopic transplantation Bcap-37 tumours of BALB/c nude mices, when tumor average volume reaches~70mm3
When, every other day (with first time injection be the 0th day) by tail vein difference injecting normal saline, pUCNPs (30mg/kg),
CPT-11 (equivalent SN38 10mg/kg) and 1@pUCNPs (equivalent SN38 10mg/kg) aqueous solution, are administered 3 times altogether, survey within every 4 days
Amount Mouse Weight and tumor size, calculate gross tumor volume：(length x width2)/2.Put to death all small within the 24th day after administration
Mouse, takes out tumour, weighs.
Fig. 9 (a) show human mammary cancer line Bcap-37 tumor bearing nude mice after drug-treated gross tumor volume with the time change
Change.The tumour growth of 1@pUCNPs administration groups has obtained substantially slowing down, and the 24th day after medication, inhibition rate of tumor growth reaches
61%, compared with physiological saline group, there is pole significant difference (n=6, p in both<0.001), the tumour of CPT-11 administration groups
It is only 14% to suppress efficiency, and physiological saline group does not exist significant difference (n=6, p>0.05).Fig. 9 (b) be tumor bearing nude mice from
The tumor quality of the 24th day compares after being administered for the first time, and 1@pUCNPs administration group tumour average qualities are significantly less than CPT-11 administrations
Group, there is significant difference (n=6, p in both<0.05).
Figure 10 shows influence of the drug-treated to Mouse Weight, and the@pUCNPs group body weight of administration initial stage 1 is declined slightly, but after
Phase increased weight shows 1@pUCNPs to animal without overt toxicity up to keeping identical growth tendency with control group.
Result shows that above-mentioned rare earth upconversion nano pharmaceutical carrier has obvious tumor growth inhibitory effect, and without bright
Aobvious toxicity, has good antitumor application thereof to be worth.
Claims (6)
1. it is a kind of to collect the fluorescence imaging rare earth upconversion nano pharmaceutical carrier integrated with medicine is carried, it is characterised in that the nanometer
Pharmaceutical carrier is supported on the anti-swollen of up-conversion nanoparticles surface by the rare earth upconversion nano particle with fluorescence imaging function
Tumor medicine and it is coated on the amphipathic nature polyalcohol molecule DSPE-PEG of nanoparticle surface
(DSPE-PEG) constitute.
2. nano-medicament carrier as claimed in claim 1, it is characterised in that the chemical group of the rare earth upconversion nano particle
As NaYF4:Yb, Ln, Ln are Er or Tm.
3. nano-medicament carrier as claimed in claim 1, it is characterised in that the antineoplastic is to have modified hydrophobicity base
One or more of combination in the 7-ethyl-10-hydroxy-camptothecin prodrug and Cabazitaxel prodrug of group.
4. nano-medicament carrier as claimed in claim 3, it is characterised in that the hydrophobic group is selected from unrighted acid
Oleic acid, linoleic acid plus linolenic acid.
5. nano-medicament carrier as claimed in claim 1, it is characterised in that the amphipathic nature polyalcohol molecule distearyl acyl group
Phosphatidyl-ethanolamine-polyethylene glycol (DSPE-PEG), molecular weight is 2000.
6. the nano-medicament carrier as described in claim 1-5 any claims is in fluorescence imaging, the application of anti-tumor aspect.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114081953A (en) * | 2021-10-19 | 2022-02-25 | 东北农业大学 | Prodrug dendrimer nano-carrier and preparation method and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102406951A (en) * | 2011-11-17 | 2012-04-11 | 苏州大学 | Thiol-polyethylene glycol modified magneto-optical composite nano-material and its application |
| CN103804472A (en) * | 2014-01-23 | 2014-05-21 | 浙江大学 | Taxane medicinal precursor |
| CN104447777A (en) * | 2014-11-26 | 2015-03-25 | 浙江大学 | Capsicine-camptothecin anti-cancer drug conjugate and preparation method and application thereof |
-
2017
- 2017-01-03 CN CN201710001962.2A patent/CN106806905B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102406951A (en) * | 2011-11-17 | 2012-04-11 | 苏州大学 | Thiol-polyethylene glycol modified magneto-optical composite nano-material and its application |
| CN103804472A (en) * | 2014-01-23 | 2014-05-21 | 浙江大学 | Taxane medicinal precursor |
| CN104447777A (en) * | 2014-11-26 | 2015-03-25 | 浙江大学 | Capsicine-camptothecin anti-cancer drug conjugate and preparation method and application thereof |
Non-Patent Citations (6)
| Title |
|---|
| CHAO WANG ET AL: "Drug delivery with upconversion nanoparticles for multi-functional targeted cancer cell imaging and therapy", 《BIOMATERIALS》 * |
| HANGXIANG WANG ET AL: "Self-Assembling Prodrugs by Precise Programming of Molecular Structures that Contribute Distinct Stability,Pharmacokinetics, and Antitumor Efficacy", 《ADV. FUNCT. MATER.》 * |
| YAN HU ET AL: "17β-Estradiol-Loaded PEGlyated Upconversion Nanoparticles as a Bone-Targeted Drug Nanocarrier", 《ACS APPL. MATER. INTERFACES》 * |
| YINGYING YUAN ET AL: "A facile supramolecular approach to fabricate multifunctional upconversion nanoparticles as a versatile platform for drug loading, in vivo delivery and tumor imaging", 《J. MATER. CHEM. B》 * |
| 宣琎楠 等: "PEG修饰NaGdF4∶Yb3+/Er3+纳米粒子合成及上转换发光性质", 《人工晶体学报》 * |
| 袁莹莹: "稀土上转换纳米颗粒制备及其在药物递送、成像及检测中的应用", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114081953A (en) * | 2021-10-19 | 2022-02-25 | 东北农业大学 | Prodrug dendrimer nano-carrier and preparation method and application thereof |
| CN114081953B (en) * | 2021-10-19 | 2023-10-17 | 东北农业大学 | Prodrug dendrimer nano-carrier and preparation method and application thereof |
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