CN106620698A - Preparation method for ZnPc-UCNP-PEG-G nano-compound - Google Patents
Preparation method for ZnPc-UCNP-PEG-G nano-compound Download PDFInfo
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
- A61K41/0057—Photodynamic therapy with a photosensitizer, i.e. agent able to produce reactive oxygen species upon exposure to light or radiation, e.g. UV or visible light; photocleavage of nucleic acids with an agent
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/535—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one oxygen as the ring hetero atoms, e.g. 1,2-oxazines
- A61K31/5375—1,4-Oxazines, e.g. morpholine
- A61K31/5377—1,4-Oxazines, e.g. morpholine not condensed and containing further heterocyclic rings, e.g. timolol
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y20/00—Nanooptics, e.g. quantum optics or photonic crystals
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y5/00—Nanobiotechnology or nanomedicine, e.g. protein engineering or drug delivery
Abstract
The invention discloses a preparation method for a ZnPc-UCNP-PEG-G nano-compound. The preparation method comprises the following steps: synthesizing NaYF4:Yb<3+> through a micro-emulsion method; modifying Er<3+> up conversion nanoparticles with polyethylene glycol through a ligand oxidizing method to obtain water-soluble UCNP-PEG-OH; modifying a photosensitive agent and bi-carboxyl polyethylene glycol onto the surface through an EDC concentration method to a obtain ZnPc-UCNP-PEG nano-compound; lastly, loading targeted molecular gefitinib (G) modified by the polyethylene glycol onto the surface to obtain the ZnPc-UCNP-PEG-G nano-compound. The ZnPc-UCNP-PEG-G prepared by the preparation method has uniform particle distribution, high crystallization performance and high photoactivity, and can be hopefully developed into an efficient and low-toxicity novel drug loading system; the PDT therapy depth of zinc phthalocyanine and targeting ability for tumor cells can be improved.
Description
Technical field
The present invention relates to biology medical material technical field, specifically a kind of ZnPc-UCNP-PEG-G nano-complexes
Preparation method.
Background technology
Optical dynamic therapy is also referred to as photochemical therapy (Photodynamic therapy, PDT), and it is referred to using specific
The light irradiation of wavelength locally or systemically injects in advance the medicine (sensitising agent, PS) of body, and sensitising agent is activated therewith and and oxygen molecule
Effect produces the cytotoxic active oxygen (ROS) of tool, through directly irreversible to tumor tissues generation with indirectly-acting
Damage.Photodynamic therapy is different from traditional treatment tumour means, and it all has alternative to target tissue and degree of injury,
The damage of normal tissue can be reduced, the very fast metabolism of energy in human body, it is to avoid toxicity, chemically stable are produced to other body parts
The advantages of property and photostability are high.But traditional PDT still suffers from some shortcomings, if sensitising agent is to tumor tissues
Targeting is not enough, simultaneously because the absorbing wavelength of sensitising agent is shorter, its tissue penetration is limited, therefore, it is larger to volume
The therapeutic effect of tumour or deep tumor is not good.Nowadays, the fast development of nanometer technology is quick to solve Photodynamic Therapy
The problems such as tumor-targeting and treatment depth of agent, provides new way.
Molecular targeted therapy refer on cellular and molecular level by fixed carcinogenic site (i.e. with tumour cell and swell
The related specific molecular target spot of tumor tissue, the target spot is probably a certain protein molecule in tumour cell, a certain nucleic acid fragment
Or a certain gene outcome), respective ligand or antibody are designed in combination with specific position, so as to effective suppressor canceration
Or blocking Cell signal propagation pathways and Tumor angiogenesis, ultimately result in death of neoplastic cells normal thin without injury
A kind of brand-new Therapeutic mode of born of the same parents.Compared with traditional chemotherapy and radiation therapeutic modality, molecular targeted therapy has very high choosing
Selecting property, also ensures that the ideal treatment result of relatively low damage normal structure while to tumour cell Efficient killing effect.Small molecule
Targeted drug Gefitinib is a kind of selective EGF-R ELISA (EGFR) tyrosine kinase inhibitor, tyrosine-kinase
Enzyme is often expressed as among the entity tumor of epithelial origin, can indirectly be suppressed by the suppression to EGFR tyrosine kinase activities
The growth of tumour cell and Angiogenesiss, and accelerate the apoptosis of tumour cell.Gefitinib（Gefitinib）It is beautiful in 2003
State FDA ratifies for treating non-small cell lung cancer, is the first listing that gets the Green Light for treating non-small cell lung cancer
(NSCLC) oral type small molecule targeted drug.Clinical test results show that Gefitinib is to colon cancer, prostate cancer, breast
The treatment such as gland cancer and head and neck neoplasm confirms effective.
Rare earth up-conversion luminescent material (upconversion luminescence nanoparticles, UCNPs) is
One kind can be excited by near infrared light (NIR) and send the luminescent material of visible ray, i.e., up-conversion nano material can be by light more
Handset system is converted into shortwave long wave, therefore is referred to as " upper conversion ".Because the luminescence process of up-conversion has run counter to this
Lentor (Stokes) law, therefore the anti-Stokes that is otherwise known as (anti-Stokes) law luminescent material.Upper conversion
Nano material has the luminosity of stable and uniqueness, and low to the toxicity of cell, its excitation wavelength is near infrared region (usually
980 nm, in the range of the light transmission window 800-1200 nm of biological tissue), it is strong to the penetrability ability of biological tissue, and
Light will not be caused to injure tissue, protein etc., while the background autofluorescence of detection can be effectively reduced, improve fluorescence noise
Than, therefore have broad application prospects at aspects such as biomarker, multi-modality imaging, medicament transport and optical dynamic therapies.
Molecular target medicine is introduced in UCNP/sensitising agent nanosystems, by preparing molecular targeted anticancer nano platform system
System, the point of penetration for making tumor pharmacother is improved to molecular level by the cellular level studied at present.The UCNP/ of synthesis is photosensitive
In agent/Gefitinib compound, the FRET effects of UCNP and sensitising agent can solve oncotherapy depth problem, sensitising agent and Ji Fei
Synergy for Buddhist nun's molecular target medicine can solve the problems, such as that targeting is not strong and medicine is short in the holdup time of tumour cell.This
A little advantages become novel molecular targeting light power carrying platform and provide greatly possibility for the design system.
The content of the invention
It is an object of the invention to provide the preparation and its application of a kind of ZnPc-UCNP-PEG-G nano-complexes, solve
Target-oriented drug is not enough in optical dynamic therapy and problem for the treatment of depth as shallow.
For achieving the above object, the present invention is adopted the following technical scheme that：
A kind of preparation method of ZnPc-UCNP-PEG-G nano-complexes, comprises the following steps that：
（1）0.4 g NaOH are weighed in 100 mL reactors, then is separately added into 20 mL deionized waters, 30 mL ethanol, 7.5
The n-hexane of the oleic acid of mL and 4 mL, stirs and evenly mixs as solution I；Weigh 0.8 mmol rare earth nitrades and be dissolved in 5 mL deionizations
In water, with 20 drop/min instill solution I solution II, then weigh 3.2 mmol NaF and be dissolved in 5 mL deionized waters and be added to
In solution II, 6 h are reacted at being placed in 180 DEG C；Reaction naturally cools to room temperature after terminating, and is extracted in three times with 30 mL hexamethylenes
Take, collect hexamethylene layer, add excess ethyl alcohol to be collected by centrifugation；Again deionized water and ethanol are alternately washed 3 times, are finally dispersed in 10
It is stand-by in mL hexamethylenes；
（2）Measure 8 mL steps（1）UCNPs cyclohexane solution, add 12 mL hexamethylenes and 10 mL dichloromethane；Again
Add 25 mg metachloroperbenzoic acids, be warming up to 54 DEG C and be stirred at reflux 3 h, be cooled to room temperature, add 1.0 g PEG-OH after
8 h of continuous reaction；After reaction terminates, decompression is spin-dried for, then alternately washes 3 times with ethanol and deionized water, and -60 DEG C lyophilized to obtain UCNP-
PEG-OH；
（3）Weigh 3 mgβ- ZnPc-COOH and 8 mg HOOC-PEG-COOH are dissolved in 5 mL DMSO and obtain mixed liquor I, then
Weigh 6 mg N, N- dicyclohexylcarbodiimides（DCC）With 4 mg DMAPs（DMAP）It is dissolved in 200 μ L's respectively
In DMSO, it is added in mixed liquor I and stirs 1 h, adds 30 mg steps（2）48 h are reacted under product, room temperature；Centrifugation is received
Collection, with DMSO 3 times are washed, and -60 DEG C lyophilized to obtain ZnPc-UCNP-PEG-COOH；
（4）By step（3）Product is dissolved in 12 mL DMSO, adds 7 mg DCC and 5 mg DMAP stirring 1h, adds compound
48 h are reacted under G, room temperature；It is collected by centrifugation, is washed 3 times with DMSO, obtains product ZnPc-UCNP-PEG-G.
Described rare earth nitrades are Y (NO3)·6H2O and Yb (NO3) ·5H2O and Er (NO3) ·5H2O is with mass ratio
68:30:2 mixing.
Step（4）The addition of middle compound G is the 5% ~ 20% of product total amount.
The compound G is the miscellaneous octyl group-Gefitinib of the alcoxyl of 8- hydroxyls -3,6.
The beneficial effects of the present invention is：
1）The present invention will synthesize ZnPc-UCNP-PEG-G nano-complexes, solve target-oriented drug deficiency in optical dynamic therapy, control
Treat the problem of depth as shallow.
2）The ZnPc-UCNP-PEG-G nano-complex uniform particle sizes of synthesis, transferring efficiency of fluorescence resonance energy is high,
980nm excites lower singlet oxygen yield high, solves the problems, such as to treat depth as shallow.
3）The ZnPc-UCNP-PEG-G nano-complexes of synthesis show superior photodynamic activity, in vitro light power
Therapeutic process has certain dose dependent to the killing ability of cell, and with the increase of concentration, killing ability strengthens, and
Obvious lethal effect is not shown to cell under non-illuminated conditions.
4) the ZnPc-UCNP-PEG-G nano-complexes of synthesis have obvious targeting to cancer cell, and normal thin
It is distributed less in born of the same parents, reduces the side effect in cancer treatment procedure.
Description of the drawings
Fig. 1 is (a) up-conversion nanoparticles NaYF4:Yb3+,Er3+(UCNPs) X-ray diffractogram；
Fig. 2 is (a) up-conversion nanoparticles NaYF4:Yb3+,Er3+(UCNPs) scheme with the TEM of (b) UCNP-PEG-OH；
Fig. 3 is the infrared spectrogram of UCNPs up-conversion nanoparticles and UCNP-PEG-OH；
Fig. 4 is NaYF4:Yb3+,Er3+(UCNPs) up-conversion luminescence figure；
I and II is the corresponding fluorescence emissions of UCNPs in Fig. 5, and III and IV is the corresponding fluorescence emissions of ZnPc-UCNP-PEG；
Fig. 6 a are DPBF, UCNPs, UCNP-PEG, β-ZnPc-COOH, ZnPc-UCNP-PEG, ZnPc-UCNP- PEG-
The Ф comparison diagrams of G；Fig. 6 b are the Ф comparison diagrams under ZnPc-UCNP-PEG-G different conditions；
Fig. 7 is HepG2 cells (circle) and HELF cells (bar shaped) to ZnPc-UCNP-PEG-G competition 24 h laser of intake
Co-focusing imaging figure；
Fig. 8 be object ZnPc-UCNP-PEG-G and tester ZnPc-UCNP-PEG to the h of HepG2 cell incubations 24 after it is glimmering
Light statistics block diagram；
Fig. 9 a and Fig. 9 b are intakes of the ZnPc-UCNP-PEG-G in cancer cell HepG2, the UCNPs of instruction andβ -ZnPc-
Being co-located at for COOH is intracellular；
The lower HepG2 cell survival rates of variable concentrations ZnPc-UCNP-PEG-G and ZnPc-UCNP-PEG incubation under Figure 10 no lights
Figure；
Figure 11 is that have the lower HepG2 cell survival rates of variable concentrations ZnPc-UCNP-PEG-G and ZnPc-UCNP-PEG incubation under illumination
Figure.
Specific embodiment
Below technical scheme is described further with specific embodiment, but protection scope of the present invention is not limited
In this:
Embodiment 1
A kind of preparation method of ZnPc-UCNP-PEG-G nano-complexes, concretely comprises the following steps：
1) 0.4 g NaOH are weighed in 100 mL reactors, then measures 20 mL deionized waters respectively, 30 mL ethanol, 7.5
The n-hexane of the oleic acid of mL and 4 mL, stirring allows it to form uniform solution.Weigh 0.8 mmol rare earth nitrades（0.2390
g Y(NO3)·6H2O, 0.1065 g Yb (NO3) ·5H2O, 0.0069 g Er (NO3) ·5H2O）It is dissolved in 5 mL deionizations
Be added dropwise in above-mentioned reactant liquor with 20 drop/min in water, then weigh 0.1314 g (3.2 mmol) NaF be dissolved in 5 mL go from
It is added in sub- water in reactant liquor, takes out stirring magneton encapsulation and be placed in baking oven at 180 DEG C and react 6 h.Reaction is natural after terminating
Room temperature is cooled to, is extracted in three times with 30 mL hexamethylenes, collect hexamethylene layer, add excess ethyl alcohol to be collected by centrifugation.Spend again
Ionized water and ethanol are alternately washed 3 times, are finally dispersed in stand-by in 10 mL hexamethylenes.
2) measure the cyclohexane solution of 8 mL UCNPs, add 12 mL hexamethylenes and 10 mL dichloromethane, then weigh 25
Mg metachloroperbenzoic acids, are warming up to 54 DEG C and are stirred at reflux after 3 h, are cooled to room temperature, add 1.0 g PEG-OH to continue anti-
Answer 8 h.After reaction terminates, decompression is spin-dried for, then alternately washes 3 times with ethanol and deionized water, and -60 DEG C lyophilized to obtain UCNP-PEG-
OH。
3) 3 mg β-ZnPc-COOH are weighed and 8 mg HOOC-PEG-COOH is dissolved in 5 mL DMSO, then weigh 6 mg
N, N- dicyclohexylcarbodiimide (DCC) and 4 mg DMAPs (DMAP) are dissolved in respectively in the DMSO of 200 μ L and add
Enter in above-mentioned reactant liquor to stir 1 h, activated carboxyl to be added and react 48 h under 30 mg UCNP-PEG- OH room temperatures.Centrifugation is received
Collection, with DMSO 3 times are washed, and -60 DEG C lyophilized to obtainβThe up-conversion nanoparticles of-ZnPc-COOH and HOOC-PEG-COOH loadings are produced
Thing is abbreviated as ZnPc-UCNP-PEG-COOH.
4) the product ZnPc-UCNP-PEG-COOH of previous step is dissolved in 12 mL DMSO and adds 7 mg DCC and 5 mg
DMAP is stirred one hour, activated carboxyl, adds 8 mg compound G, and 48 h are reacted under room temperature.It is collected by centrifugation, is washed 3 times with DMSO,
The lyophilized product for obtaining is abbreviated as ZnPc-UCNP-PEG-G.
The compound G is the miscellaneous octyl group-Gefitinib of the alcoxyl of 8- hydroxyls -3,6.
Application Example 1
By the research of ZnPc-UCNP-PEG-G nano-complex singlet oxygen yields, the tissue penetration of its light is studied,
Reference is provided for research treatment depth, with more important meaning.
ZnPc-UCNP-PEG-G nanometer systems are existed using the diphenyl isobenzofuran (DPBF) of chemical probe 1,3
The generation ability of the lower singlet oxygen of 980 nm laser irradiation is detected.Specific experiment method of operating is：Weigh a small amount of DPBF
Fully dissolved with DMSO, then adjusting makes its absorbance at 420 nm be 1, while adjusting ZnPc-UCNP-PEG-G's
Absorbance of the DMSO solution at 670 nm is also 1.Take 1 mL DMSO solvents and 1 mL ZnPc-UCNP- PEG-G it is molten
Liquid is sufficiently mixed in cuvette, and as reference systematic error, then the DMSO solution for taking DPBF and ZnPc-UCNP-PEG-G are eliminated
Each 1 mL is well mixed, and determines its absorbance at 420 nm, now for 0 s when DPBF absorbance, then hard
Rate density is 1 W/cm2980 nm laser illumination cuvette in mixed solution, often irradiate 30 s and survey absorbance,
Survey 10 time points.Respectively to DPBF, UCNPs, UCNP-PEG,β- ZnPc-COOH, ZnPc-UCNP-PEG, ZnPc-UCNP-
PEG-G solution1O2Measure.Test result is as shown in figures 6 a and 6b.Add the tissue of one layer of 1 cm thickness in cuvette mouth,
With the laser of 670 nm and 980 nm wavelength respectively determining ZnPc-UCNP-PEG-G'sФSinglet oxygen yield, and with not plus
That what is organized compares.
Can be seen that after the tissue coverage for having added 1 cm by Fig. 6 a and Fig. 6 b experimental datas, with the list of 980 nm light irradiations
Line state oxygen yield only have dropped 40.68 %, and have dropped 88.9 % with the singlet oxygen yield of 670 nm light irradiations.Therefore, with
Directly excited with the light of 670 nmβ- ZnPc-COOH is compared, and is excited indirectly with the light of 980 nmβ- ZnPc-COOH have penetrate
The advantage of thick tissue, has further highlighted the applicability that ZnPc-UCNP-PEG-G is treated in entity or deep tumor.
Application Example 2
Can medicine effectively kill tumour cell or cause certain damage with tumour cell to medicine to tumour cell
Intake has very big relation, therefore, research cell is necessary to the intake of medicine
The HepG2 cells in exponential phase are chosen, after PBS washings pancreatin digestion process is added, adding culture medium will
Cell piping and druming is uniform, adds new culture medium again to blow and beat cell uniformly after centrifugation, and cell is counted, will with culture medium
Cell is diluted to 300,000 cell/mL, adds with 100 μ L per hole in 96 orifice plates, and each medicine sets 6 multiple holes, and cell is placed in
Overnight growth in cell culture incubator.Old culture medium is discarded after adherent, 100 μ L medicines-culture medium solution (medicine is added per hole
Mother liquor is 64 mg/ mL, and solvent is DMSO；The final adding consistency of the event of DMSO amounts≤1% is 640 μ g/ in order to ensure culture medium
ML), it is placed in incubator and is incubated 24 h.Upper strata culture medium is discarded, is washed with PBS three times, thoroughly to remove the medicine not absorbed,
Then the SDS lysates for adding 120 μ L concentration to be 1% in every hole, are placed on 37oC shaking tables rock 30 min of cracking, with
The light of the excitation wavelength of relative medicine is excited, and with multi-function microplate reader its fluorescence intensity at maximum emission wavelength is detected.
Meanwhile, do the calibration curve (8 concentration, 2 multiple holes) of relative medicine：With SDS it is solvent from 325 μ by medicine
Successively twice dilution obtains eight variable concentrations to g/ mL, and with the same terms respective fluorescence intensity is determined, soft with Origin 8.5
Part process, does fluorescence intensity-concentration rectilinear, obtains final product the calibration curve of respective medicine.By fluorescence intensity Jing in each hole of experimental group
Pharmaceutical standards curve is obtained each hole cellular uptake drug concentration.
Using BCA protein quantification detection methods, (direct result of calculation need to expand 6 times, as to calculate the protein content in each hole
The value of each experimental group).Again with the drug concentration in each hole divided by each PFP total amount, its ratio is then contained in unit albumen
The amount of sensitising agent.Block diagram (Fig. 8) is done to individual ratio with the statistical analysis softwares of Graph Pad Prism 5.0, as a result with Mean
± SEM represents, parallel laboratory test in triplicate under the same terms.
Be connected to it is polyethyleneglycol modified after the ZnPc-UCNP-PEG-G that obtains of targeted molecular Gefitinib (G) exist
Intake in HepG2 cells illustrates ZnPc-UCNP-PEG-G far above the ZnPc-UCNP-PEG for not connecting Gefitinib
There is good targeting to hepatocellular carcinoma H22.
Application Example 3
Cytotoxicity test mainly studies the security of Photodynamic Therapy, and toxic action of the medicine to cell.
The good HepG2 cells for being in logarithmic phase of growth conditions are taken, after washing, digestion process, cell is carried out
Count, it is 100,000 cell/mL that the cell for having counted is diluted to into concentration, be then inoculated with 100 μ L per hole in 96 orifice plates, will be thin
Incubated overnight in born of the same parents' incubator.ZnPc-UCNP-PEG-G and ZnPc-UCNP-PEG are prepared into 7 concentration gradients with DMSO solution
Mother liquor, then with culture medium by 100 times of each concentration dilution.Jing after PBS washs 3 times, the drug solution after 100 μ L dilutions is taken
In adding each hole into 96 orifice plates, experimental group sets 6 multiple holes.Blank control group and cell controls group and experimental group are set,
Dosing is placed in be put in cell culture incubator after finishing and is incubated 24 h.
Dark poison experiment：After the h of cell incubation 24 that agent-feeding treatment before is crossed, old culture medium, with PBS 3 are washed before discarding
Time, then 100 μ L culture mediums are added in every hole, it is placed in incubator and is incubated 24 h.
Light poison experiment：After being incubated 24 h, old culture medium, is washed 3 times with PBS before discarding, then adds 100 μ in every hole
The new culture mediums of L, (power density is 1.0 W/cm to irradiate 20 min with 980 nm LEDs2) after, then 96 orifice plates are placed in into 37 °
C constant temperature and 5% CO2Continue to be incubated 24 h in culture medium.
OD values are detected：The MTT solution (5 mg/mL) that the complete cell per well 10 μ L of addition of above-mentioned incubation have been prepared, is placed in
4 h are incubated in incubator, liquid original in orifice plate is discarded, then 100 μ L DMSO are added in every hole, be placed in concussion half in shaking table
Hour or so, after first a ceremonial jade-ladle, used in libation is completely dissolved, determine in 96 orifice plates per OD value of the hole at 570 nm.By acquired results Graph
Pad Prism 5.0 are processed, and are as a result represented with Mean ± SEM.
Test result indicate that ZnPc-UCNP-PEG-G nano-complexes have significantly to kill to hepatocellular carcinoma H22 making
With, and kill ability there is certain dose dependent, with the increase of concentration, killing ability strengthens, and Figure 10 is in no light
Under the conditions of ZnPc-UCNP-PEG to HepG2 cells without obvious lethal effect, and ZnPc-UCNP-PEG-G is in certain concentration model
Enclose it is interior to HepG2 cells still have certain lethal effect, this explanation Gefitinib introducing improve target-oriented drug
The Chemotherapy of Gefitinib is also achieved simultaneously, realizes the dual activity of targeting PDT and chemotherapy.
The foregoing is only presently preferred embodiments of the present invention, all impartial changes done according to scope of the present invention patent with
Modification, should all belong to the covering scope of the present invention.
Claims (5)
1. a kind of preparation method of ZnPc-UCNP-PEG-G nano-complexes, it is characterised in that：Comprise the following steps that：
（1）The preparation of upper conversion particles UCNPs：0.4 g NaOH are weighed in 100 mL reactors, add 20 mL go from
The n-hexane of sub- water, 30 mL ethanol, the oleic acid of 7.5 mL and 4 mL, stirs and evenly mixs as solution I；Weigh 0.8 mmol rare earth nitre
Hydrochlorate is dissolved in 5 mL deionized waters, with 20 drop/min instill solution I solution II, then weigh 3.2 mmol NaF and be dissolved in
It is added in solution II in 5 mL deionized waters, 6 h is reacted at being placed in 180 DEG C；Reaction naturally cools to room temperature after terminating, and uses
30 mL hexamethylenes are extracted in three times, collect hexamethylene layer, add excess ethyl alcohol to be collected by centrifugation；Again deionized water and ethanol are handed over
For washing 3 times, conversion particles UCNPs are obtained, be finally dispersed in stand-by in 10 mL hexamethylenes；
（2）The preparation of UCNP-PEG-OH：Measure 8 mL steps（1）UCNPs cyclohexane solution, add 12 mL hexamethylenes
With 10 mL dichloromethane；25 mg metachloroperbenzoic acids are added under stirring, 54 DEG C is warming up to and is stirred at reflux 3 h, be cooled to
Room temperature, adds 1.0 g PEG-OH to continue to react 8 h；After reaction terminates, decompression is spin-dried for, then is alternately washed with ethanol and deionized water
3 times, -60 DEG C lyophilized to obtain UCNP-PEG-OH；
（3）The preparation of ZnPc-UCNP-PEG-COOH：Weigh 3 mg β-ZnPc-COOH and 8 mg HOOC-PEG-COOH are dissolved in 5
Mixed liquor I is obtained in mL DMSO, then weighs 6 mg N, N- dicyclohexylcarbodiimides and 4 mg DMAPs are distinguished
In being dissolved in the DMSO of 200 μ L, it is added in mixed liquor I and stirs 1 h, adds 30 mg steps（2）Product, reacts under room temperature
48 h；It is collected by centrifugation, with DMSO 3 times is washed, -60 DEG C lyophilized obtains ZnPc-UCNP-PEG-COOH；
（4）The preparation of ZnPc-UCNP-PEG-G nano-complexes：By step（3）Product is dissolved in 12 mL DMSO, adds 7 mg
N, N- dicyclohexylcarbodiimide and 5 mg DMAPs stirring 1h, add compound G, and 48 h are reacted under room temperature；From
The heart is collected, and is washed 3 times with DMSO, obtains product ZnPc-UCNP-PEG-G.
2. the preparation method of ZnPc-UCNP-PEG-G nano-complexes according to claim 1, it is characterised in that：Rare earth
Nitrate is Y (NO3)·6H2O and Yb (NO3) ·5H2O and Er (NO3) ·5H2O is with mass ratio 68:30:2 mixing.
3. the preparation method of ZnPc-UCNP-PEG-G nano-complexes according to claim 1, it is characterised in that：Step
（4）The addition of middle compound G is the 5% ~ 20% of product total amount.
4. the preparation method of a kind of ZnPc-UCNP-PEG-G nano-complexes according to claim 1, it is characterised in that：
Compound G is the miscellaneous octyl group-Gefitinib of the alcoxyl of 8- hydroxyls -3,6.
5. a kind of application of the ZnPc-UCNP-PEG-G nano-complexes of preparation as claimed in claim 1, it is characterised in that：
Described ZnPc-UCNP-PEG-G nano-complexes are applied in optical dynamic therapy medicine.
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Cited By (4)
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| CN108452304A (en) * | 2018-03-13 | 2018-08-28 | 浙江大学 | The preparation method of conversion composite nano materials and product and application on rare earth |
| CN110960686A (en) * | 2019-12-26 | 2020-04-07 | 中国医学科学院生物医学工程研究所 | Phthalocyanine compound for tumor targeted fluorescence imaging and photodynamic therapy |
| CN111892922A (en) * | 2020-09-01 | 2020-11-06 | 哈尔滨工程大学 | Preparation method of rare earth up-conversion nanoparticle/bismuth vanadate nanocomposite material with anti-tumor effect |
| CN115487296A (en) * | 2022-09-23 | 2022-12-20 | 青岛科技大学 | Phthalocyanine self-assembly nano-particles for tumor diagnosis and treatment and preparation method thereof |
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| CN108452304B (en) * | 2018-03-13 | 2020-08-11 | 浙江大学 | Preparation method of rare earth up-conversion composite nano material, product and application |
| CN110960686A (en) * | 2019-12-26 | 2020-04-07 | 中国医学科学院生物医学工程研究所 | Phthalocyanine compound for tumor targeted fluorescence imaging and photodynamic therapy |
| CN111892922A (en) * | 2020-09-01 | 2020-11-06 | 哈尔滨工程大学 | Preparation method of rare earth up-conversion nanoparticle/bismuth vanadate nanocomposite material with anti-tumor effect |
| CN111892922B (en) * | 2020-09-01 | 2021-05-14 | 哈尔滨工程大学 | Preparation method of rare earth up-conversion nanoparticle/bismuth vanadate nanocomposite material with anti-tumor effect |
| CN115487296A (en) * | 2022-09-23 | 2022-12-20 | 青岛科技大学 | Phthalocyanine self-assembly nano-particles for tumor diagnosis and treatment and preparation method thereof |
| CN115487296B (en) * | 2022-09-23 | 2023-08-08 | 青岛科技大学 | Phthalocyanine self-assembled nano-particles for tumor diagnosis and treatment and preparation method thereof |
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