CN111760024B - Permeation enhanced gold nanocluster drug-loaded targeting preparation and preparation method and application thereof - Google Patents
Permeation enhanced gold nanocluster drug-loaded targeting preparation and preparation method and application thereof Download PDFInfo
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- CN111760024B CN111760024B CN202010723241.4A CN202010723241A CN111760024B CN 111760024 B CN111760024 B CN 111760024B CN 202010723241 A CN202010723241 A CN 202010723241A CN 111760024 B CN111760024 B CN 111760024B
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Abstract
The invention discloses a penetration enhanced gold nanocluster drug-loaded targeting preparation as well as a preparation method and application thereof, wherein the targeting preparation comprises gold nanoclusters, a cross-linking agent and a tumor treatment drug; the gold nanoclusters are obtained by assembling gold nanospheres with a cross-linking agent, and targeted tumor treatment drugs of human epidermal growth factor receptor-2 are modified on the surfaces of the gold nanoclusters. The preparation can be used as an anti-tumor drug carrier, specifically targets to tumor, efficiently releases the drug at the tumor part through tumor microenvironment reduction and NIR irradiation, combines thermotherapy and action, and improves the treatment effect. The preparation can also combine the advantages of gold in CT imaging, photoacoustic imaging (PA) imaging and other imaging aspects to carry out medical diagnosis, thereby achieving the aim of multi-mode tumor accurate diagnosis and treatment.
Description
Technical Field
The invention relates to the field of medicinal preparations, in particular to a penetration enhanced gold nanocluster medicine-carrying targeted preparation and a preparation method and application thereof.
Background
Gold nanospheres refer to associative colloids with a diameter of between 1 and 150nm, typically prepared by reducing chloroauric acid. The gold nanometer has better biocompatibility, the surface of the gold nanometer is easy to be combined with various medicines or macromolecules in the modes of electrostatic adsorption, covalent or non-covalent combination and the like, and the gold nanometer can be used as a carrier for a medicine delivery system. In addition, the gold nanoparticles have high electron density, dielectric property and optical property, can convert absorbed light energy into heat energy through Surface Plasmon Resonance (SPR), and can be applied to the research fields of cancer diagnosis, biological probes, photothermal therapy, radiotherapy sensitization and the like.
Photothermal therapy (HT) is a method of treating tumors by artificially raising the temperature of human tissues. Currently, photo-thermal methods such as gold nanoshells, gold nanorods and gold nanocages are used for realizing cancer treatment, which draws wide attention. These gold nanoparticles absorb light in the Near Infrared (NIR) region (700-900nm) and can therefore exhibit NIR radiation-induced photothermal effects. By engineering the morphology of gold nanoparticles, their LSPR can be tuned to Near Infrared (NIR) wavelengths. Near infrared radiation can penetrate body tissue and blood to a depth of a few millimeters, making these particles useful for imaging and photothermal therapy. And (3) using the liposome as a template, and carrying out surface in-situ reduction to generate gold nanoclusters so as to form a biodegradable plasma resonance structure. After NIR irradiation, the temperature sensitive lipid membrane is destroyed and the gold shell is degraded into 5-8nm nanoparticles. Hydrophilic or hydrophobic drugs can be encapsulated within these gold-coated liposomes and the drug can be released with NIR triggering, and the gold nanoparticles produced after NIR irradiation can be cleared by the kidneys.
Antibody-mediated molecular-targeted therapy (antibody-mediated molecular-targeted therapy) is a novel drug therapy mode aiming at specific biological pathways of tumor development with the deep understanding of the occurrence mechanism and biological characteristics of tumors. Among them, antibody therapy has achieved clinical success in the treatment of hematological malignancies and solid tumors by improving efficacy and reducing toxicity. Antibody-mediated molecular targeted therapy is rapidly developed by linking antibodies to biologically active molecules to form antibody-drug conjugates. Antibody targeting has also been widely utilized by conjugating antibodies on the nanoparticle surface for efficient delivery to target sites (tumors) to improve therapeutic index and minimize off-target side effects in normal tissues. Among them, Trastuzumab (Trastuzumab), the trade name Herceptin (Herceptin), was approved by FDA in the united states in 1998, is a targeted therapeutic against the HER2 target of breast cancer, shows therapeutic effects in the treatment of both early and advanced (metastatic) breast cancer, and has advantages in that it retains the high affinity of HER2 receptor and solves the problem of immunogenicity of murine monoclonal antibodies to human.
Medical imaging technology plays an important role in disease treatment, such as early diagnosis of disease, real-time imaging during surgery, and effect tracking after treatment, and the like, and can greatly improve the cure rate of disease, but a single imaging mode cannot completely provide morphological and functional information of tumor due to the limitation of each imaging mode. Therefore, the development of the multi-modal imaging technology can fully play the advantages of each imaging mode and greatly improve the precision of tumor diagnosis. Photoacoustic imaging (PA) is an emerging imaging technology, in which after laser irradiation, biological tissues convert light energy into heat energy to increase the local temperature, and pressure waves are generated due to thermal expansion to obtain Photoacoustic signals, which is a conversion process from "light energy" - "heat energy" - "mechanical energy". The photoacoustic imaging has the advantages of no wound, simple and convenient operation, high resolution and contrast and the like. Photoacoustic imaging includes endogenous and exogenous contrast imaging, endogenous utilizing endogenous chromophore in biological tissue, such as melanin, hemoglobin, etc., however, biological tissue itself has weak light absorption, and exogenous contrast agent is usually introduced to enhance contrast. Due to the surface plasmon resonance effect of the gold nano, when the gold nano is irradiated by light with proper wavelength, the light energy can be converted into heat energy to generate a photoacoustic signal, the photoacoustic conversion efficiency is high, the light absorption of an endogenous chromophore can be effectively reduced, the imaging depth is maximized, and the gold nano-composite material is widely applied to the photoacoustic imaging research field.
The nanoparticles are widely applied to tumor targeting research due to the unique properties of large specific surface area, easy modification and the like, and in recent years, research on nano delivery carriers mainly focuses on how to design nano carriers with good targeting property, high drug-loading rate and the like, however, after the carriers reach tumor sites through EPR effect or active targeting effect, how to realize effective retention of the carriers at the tumor sites and penetration of the carriers in deep tumor sites are more and more concerned by researchers. It has been found that the effect of drug delivery to the tumor site alone is far from sufficient, and penetration of the tumor site greatly affects the therapeutic effect. At present, factors influencing aggregation and penetration of the drug in the tumor are mainly influenced by physicochemical characteristics of the nano-drug, such as size, shape and charge of particles and physiological barriers, wherein the size of the nano-drug is a key factor for the aggregation and penetration of the nano-drug at the tumor site. Firstly, the nano-drugs can reach tumor sites by effectively utilizing EPR effect only in a proper particle size range, numerous literature researches indicate the correlation between the particle size and the penetration and retention of nano-particles in tumors, the nano-drugs with larger sizes are not easy to penetrate to the deep part of the tumors through diffusion, so that the nano-drugs cannot be uniformly distributed in the tumors, and the nano-drugs with smaller sizes cannot be effectively retained because the nano-drugs with smaller sizes are easy to externally permeate back to blood circulation due to interstitial high pressure. Therefore, the realization of the size adjustability of nanoparticles is one of the development directions of the future nano-drug research.
Currently, patents related to the application of gold nanoclusters or gold nano assemblies include: a gold nanocluster and a preparation method and application thereof (publication number: CN 110935030A); ② a fluorescent gold nano-cluster and a preparation method and application thereof (publication No. CN 111253930A); and thirdly, a preparation method and application (publication number: CN106924764A) of a functional photoacoustic probe based on gold nanoparticle assembly. The gold nanoclusters described in the patents I and II are different from the gold nanoclusters disclosed by the invention, are a subminiature gold nanoparticle (less than 5nm) aggregate, have fluorescence, and can be used as a drug carrier and a fluorescent probe for treating and detecting diseases. However, the gold nanoparticles have too small particle size, and do not have the characteristics of photothermal effect, photoacoustic imaging and the like of the gold nanoparticles. The gold nanoparticle assembly is an assembly of gold nanorods and gold nanospheres, and is used for photoacoustic imaging. However, the gold nanorods have the characteristics of large toxicity and difficult metabolism, so that the prospect of the gold nanorods in clinical application is limited. Compared with the invention, the polypeptide-assembled gold nanocluster provided by the invention creatively utilizes the tumor microenvironment to realize the disintegration of the gold nanocluster on the basis of keeping the gold nanocluster photothermal effect, so that the gold nanocluster is used as a drug carrier to mediate combination therapy and photoacoustic imaging, and meanwhile, the metabolizability of the carrier is considered. In a word, the gold nanocluster assembled by the polypeptide has the advantages of simplicity in synthesis, easiness in combination treatment and imaging, easiness in degradation and the like, and has great application and research values in the field of medicines.
Disclosure of Invention
The purpose of the invention is as follows: the invention aims to provide a polypeptide-assembled gold nanocluster drug-loaded targeting preparation. The invention also aims to provide a preparation method of the preparation. The invention also aims to indicate that the preparation can be used as a good anti-tumor drug carrier and can be applied to multi-mode thermosensitive immune precise diagnosis and treatment of tumors.
The invention is realized by the following technical scheme:
a penetration enhanced gold nanocluster drug-loaded targeting preparation is characterized in that the gold nanocluster drug-loaded targeting preparation comprises gold nanoclusters, a cross-linking agent and a tumor treatment drug; the gold nanoclusters are obtained by assembling gold nanospheres with a cross-linking agent, and targeted tumor treatment drugs of human epidermal growth factor receptor-2 are modified on the surfaces of the gold nanoclusters.
Further, the grain diameter of the gold nanocluster is 5-15 nm, and the potential is-20 mv to-40 mv; the particle size of the penetration enhanced gold nano-cluster drug-loaded targeting preparation is 50-80 nm, the potential is-20-40 mv, and the ultraviolet maximum absorption wavelength of the penetration enhanced gold nano-cluster drug-loaded targeting preparation is 500-600 nm.
Further, the tumor therapeutic drug is a tumor therapeutic agent selected from a small molecule chemotherapeutic agent, an inorganic metal therapeutic drug, a protein therapeutic drug or a gene therapeutic drug.
Further, the cross-linking agent for assembling is polypeptide, polyethylene glycol and dendritic polymer with two sulfhydryl groups, and the cross-linking is gold-sulfur bond covalent bonding or electrostatic adsorption;
the thiol-terminated polypeptide is preferably a thiol-terminated matrix metalloproteinase-sensitive polypeptide.
Further, the small molecule chemotherapeutic agent is selected from one or more of maytansine and its derivatives, doxorubicin hydrochloride, camptothecin, epothilone, taxane, docetaxel, daunorubicin, doxorubicin, epirubicin, mitoxantrone, idarubicin, cytarabine, erlotinib, afatinib, lapatinib, dactinotinib, gefitinib, AP32788, poetinib, ocitinib, EGF816, gemitinib, crilaitinib, tandatinib, sorafenib, midostaurin, quinatinib, pemetrexed; preferred maytansine derivatives are DM1(CAS No.: 139504-50-0);
the protein therapeutic drug or the tumor therapeutic agent of the gene therapeutic drug is selected from one or more of trastuzumab, AMP-224, AMP-514/MEDI-0680, astuzumab, avizumab, BGB-A317, BMS936559, Devacizumab, JTX-4014, nivolumab, pembrolizumab and SHR-1210; preferably trastuzumab;
the inorganic metal therapeutic drugs are gold, iron, bismuth and platinum therapeutic drugs; carboplatin or cisplatin is preferred.
The invention also provides a preparation method of the permeation enhanced gold nanocluster drug-loaded targeting preparation, which comprises the following steps:
(1) reducing sodium borohydride to prepare gold nanospheres;
(2) crosslinking the gold nanospheres to prepare gold nanoclusters;
(3) the gold nanocluster reacts with the tumor treatment drug to prepare the gold nanocluster drug-carrying targeting preparation.
Further, the preparation method comprises the following steps:
respectively measuring 10-25mM gold chloride acid 100-; taking 1-1.5mL of gold nanosphere, adding 1-2mL of polypeptide (CRDPLGLAGDRC) solution with the concentration of 1-2mg/mL, gently rotating until the mixture is uniformly mixed, stirring for 12-24 hours at room temperature, centrifuging by using an ultrafiltration tube with the molecular weight cutoff of 5-15kDa to obtain a gold nanocluster assembled by polypeptide, adding 1-2mg/mL of maytansine derivative DM1 solution, stirring for 12-24 hours at room temperature, adding an antibody solution dissolved in PBS, gently oscillating for 12-24 hours at room temperature, and centrifuging by using an ultrafiltration tube with the molecular weight cutoff of 300-500kDa to obtain a gold nanocluster drug-carrying targeting preparation assembled by polypeptide.
Further, the preparation method comprises the following steps:
(1) respectively measuring 10-25mM chloroauric acid 100-;
(2) taking 1-1.5mL of the gold nanospheres prepared in the step (1), adding 1-2mL of polypeptide solution with the concentration of 1-2mg/mL, slightly rotating until the mixture is uniformly mixed, stirring at room temperature for 12-24 hours, and centrifuging by using an ultrafiltration tube with the molecular weight cutoff of 5-15kDa to obtain polypeptide-assembled gold nanoclusters;
(3) adding 1-2mg/mL maytansine derivative DM1 solution into the polypeptide-assembled gold nanoclusters prepared in the step (2), stirring at room temperature for 12-24 hours, and centrifuging by using an ultrafiltration tube with the molecular weight cutoff of 5-15kDa to obtain a polypeptide-assembled gold nanocluster drug-carrying preparation;
(4) and (4) adding the polypeptide-assembled gold nanocluster drug-loaded preparation prepared in the step (3) into an antibody solution dissolved in PBS, slightly oscillating for 12-24 hours at room temperature, and centrifuging by using an ultrafiltration tube with the molecular weight cutoff of 300-500kDa to obtain the polypeptide-assembled gold nanocluster drug-loaded targeting preparation.
The invention also provides application of the penetration enhanced gold nanocluster drug-loaded targeting preparation in preparation of drugs for treating breast cancer, melanoma and prostate cancer malignant tumors.
The invention also provides application of the permeation enhanced gold nanocluster drug-loaded targeting preparation in preparation of combined radiotherapy, phototherapy and thermal therapy diagnostic reagents.
The invention also provides application of the permeation enhanced gold nanocluster drug-loaded targeting preparation in preparation of diagnostic reagents for electron Computed Tomography (CT), Magnetic Resonance Imaging (MRI) and photo-acoustic imaging (PA).
The constructed penetration enhanced gold nanocluster drug-loaded targeting preparation is disassembled under the action of matrix metalloproteinase highly expressed in a tumor microenvironment, the gold nanoclusters are cracked into small-sized gold nanoparticles, the size and form transformation of NPs (N-phospho peptides) is selectively realized in the tumor microenvironment, and the characteristics of large particle pharmacokinetics, strong tumor enrichment capacity and strong small particle penetrating capacity can be combined, so that the tissue permeability of the nanoparticles is improved by means of the preparation, and the nanoparticles can be deeply treated in tumors.
The plasma resonance nano structure with good biocompatibility and degradation capability, namely the gold nano cluster drug-loaded targeting preparation assembled by the polypeptide, can promote the gold nano cluster to control and release the drug under the condition of a tumor microenvironment, and the small-particle-size gold nano particles generated after disintegration are easy to remove by kidneys, so that the multi-mode cooperative treatment strategy combining chemotherapy and phototherapy has small metal accumulated toxicity, and the tumor treatment effect can be further improved.
The invention adopts electrostatic adsorption to connect the antibody HER2 and the gold nanocluster drug-loaded preparation assembled by the polypeptide. Due to the effective connection of the HER2 antibody, breast cancer tissues can be selectively targeted, the penetration in tumors can be enhanced, the synergistic treatment effect of the antibody drug and the carrier drug in combination therapy can be synergized, and the metabolism of the nano carrier in vivo is facilitated so as to improve the safety of the application of the carrier in vivo.
The gold nanocluster drug-loaded targeting preparation assembled by the polypeptide can be used for photoacoustic imaging by utilizing the stronger Surface Plasmon Resonance (SPR) effect and higher optical absorption coefficient of colloidal gold, so that precise diagnosis of superficial malignant tumors such as breast cancer and the like is realized through the photoacoustic imaging effect of the gold nanoclusters assembled by the polypeptide.
Has the advantages that: the penetration enhanced gold nanocluster drug-carrying targeting preparation constructed by the invention integrates the radiotherapy and thermotherapy effects and can realize a visible targeting tumor gold nanocluster diagnosis and treatment system of degradation and metabolism in vivo. The preparation can promote penetration, improve the tumor targeting treatment effect, reduce the toxic and side effects of single-means treatment, and has higher medical diagnosis and treatment application value. The preparation can be used as a good antitumor drug carrier, the stable connection of the antibody can be specifically targeted to a tumor, the drug can be efficiently released at the tumor part through NIR irradiation, and the therapeutic effect is improved by combining the effects of thermotherapy.
Drawings
Fig. 1 is a particle size diagram of gold nanospheres (a), polypeptide-assembled gold nanoclusters (B), and polypeptide-assembled gold nanocluster drug-loaded targeting agents (C) according to the present invention;
FIG. 2 is a transmission electron microscope image of gold nanospheres (A), polypeptide-assembled gold nanoclusters (B) and polypeptide-assembled gold nanocluster drug-loaded targeting preparation (C);
FIG. 3 is a UV full wavelength scan of gold nanospheres (A) and polypeptide-assembled gold nanoclusters (B);
FIG. 4 is a full wavelength scan ultraviolet of a polypeptide-assembled gold nanocluster drug-loaded targeting formulation;
FIG. 5 is a photo-thermal conversion temperature-raising curve of the gold nanocluster drug-loaded targeting preparation assembled by polypeptides (GNPs are gold nanospheres; GNCs are gold nanoclusters assembled by polypeptides; HER-GNCs are gold nanocluster targeting preparations assembled by polypeptides; HER-GNCs-DM1 are gold nanocluster drug-loaded targeting preparations assembled by polypeptides);
FIG. 6 is the in vitro release curve of gold nanocluster drug-loaded targeting preparation assembled by polypeptide with/without GSH/NIR/MMP-9 (DM1 is free maytansine derivative drug; GNCs-DM1 is gold nanocluster loaded with DM 1; HER-GNCs-DM1 is gold nanocluster drug-loaded targeting preparation assembled by polypeptide; GNCs-DM1, w/GSH is GSH treated group of gold nanocluster loaded with DM 1; GNCs-DM1, w/GSH + NIR is GSH and NIR treated group of gold nanocluster loaded with DM 1; HER-GNCs-DM1, w/GSH is GSH treated group of gold nanocluster drug-loaded targeting preparation assembled by polypeptide; HER-GNCs-DM1, w/GSH + NIR is gold nanocluster targeted targeting preparation assembled by polypeptide, GSH and NIR treated group of HER-GNCs-1, w/GSH + MMP + 9 is nano cluster drug-loaded targeting preparation assembled by polypeptide, NIR and MMP-9 treatment groups);
FIG. 7 is a transmission electron microscope image of the gold nanocluster drug-loaded targeting preparation assembled by the polypeptide, which is disintegrated after being treated by MMP-9 for 0 hour, 2 hours and 24 hours;
FIG. 8 shows the survival rates of SK-BR-3 and MCF-7 cells of free maytansine derivatives as tumor therapeutic agents;
FIG. 9 shows the survival rates of SK-BR-3(A) and MCF-7 (B) cells of the gold nanocluster drug-loaded targeting preparation assembled by polypeptides (GNCs-DM1 is a gold nanocluster loaded with DM 1; HER-GNCs-DM1 is a gold nanocluster drug-loaded targeting preparation assembled by polypeptides; HER-GNCs-DM1+ NIR is the combination of the gold nanocluster drug-loaded targeting preparation assembled by polypeptides and NIR);
FIG. 10 shows the permeability of gold nanocluster preparations in 3D tumor spheres (SH-PEG-FITC is FITC modified by mercaptopolyethylene glycol; FITC-HGNPs are FITC labeled non-enzyme-responsive hollow gold nanoparticles, gold nanoclusters labeled by FITC-GNCs; FITC-HER-GNCs are FITC labeled antibody-modified gold nanoclusters;
FITC-HER-GNCs + NIR are FITC-labeled antibody-modified gold nanoclusters irradiated by NIR);
FIG. 11 shows in vivo PA imaging of polypeptide-assembled gold nanocluster drug-loaded targeting formulations (Saline is physiological Saline, GNCs is polypeptide-assembled gold nanoclusters, and HER-GNCs is polypeptide-assembled gold nanocluster targeting formulations).
Detailed Description
The technical solutions of the present invention are further illustrated by the following specific embodiments, but it is easily understood by those skilled in the art that the specific material ratios, process conditions and results thereof described in the examples are only for illustrating the present invention, and should not also limit the present invention described in detail in the claims.
Example 1: preparation and characterization of Gold Nanoclusters (GNCs) assembled by Gold Nanospheres (GNPs) and polypeptides
Taking 100-200 mu L of chloroauric acid solution with the concentration of 20-30 mM and sodium citrate solution respectively, adding ultrapure water to dilute the chloroauric acid solution and the sodium citrate solution to a 50mL round-bottom flask until the total volume is 20mL, quickly stirring the chloroauric acid solution and the sodium citrate solution, taking ice water to prepare 50-100 mM sodium borohydride solution, quickly adding the sodium borohydride solution into the round-bottom flask with the volume of 0.5-1 mL, quickly stirring the sodium borohydride solution for 1-2 hours at room temperature to obtain gold nano solution, centrifuging the gold nano solution by using an ultrafiltration tube with the molecular weight cutoff of 10-25 kDa to remove unreacted micromolecules and concentrating the unreacted micromolecules to obtain gold nanosphere GNPs solution, and storing the gold nanosphere GNPs solution in a refrigerator at 4 ℃ for later use.
1 to 2 x 10 of-9Weighing 2.5-3.5 mg of polypeptide (CRDPLGLAGDRC) and dissolving in a proper amount of ultrapure water, dropwise adding into the gold nanoparticle solution under the protection of nitrogen, continuously bubbling for a plurality of minutes by using nitrogen, stirring overnight at room temperature, centrifugally washing by using an ultrafiltration tube with the molecular weight cutoff of 10-25 kDa to obtain a gold nanocluster GNCs solution, and storing in a refrigerator at 4 ℃ for later use.
The particle size of the gold nanospheres and the gold nanocluster solution was measured by a Zetaplus laser particle size analyzer, taking 3mL each.
Taking a proper amount of prepared gold nanospheres and gold nanocluster solution, completely wetting the copper mesh covered with the carbon film for 30s, then sucking off redundant liquid by using filter paper, drying for about 3min under light, and observing the shape and particle size by using a transmission electron microscope.
Taking 3mL of each gold nanosphere and gold nanocluster solution, and scanning the ultraviolet spectrum of the gold nanosphere and the gold nanocluster solution by using an ultraviolet-visible spectrophotometer within the range of 400-900 nm. The results are shown in FIGS. 1, 2 and 3.
Example 2: preparation of gold nanocluster targeting preparation (HER-GNCs) assembled by polypeptide and preparation and characterization of gold nanocluster drug-loaded targeting preparation (HER-GNCs-DM1) assembled by polypeptide
And (3) adjusting the pH value of the prepared gold nanocluster GNCs solution to 6-7, adding an HER2 antibody solution dissolved in PBS, lightly oscillating overnight at room temperature, centrifuging by using an ultrafiltration tube with the molecular weight cutoff of 300-500kDa to obtain an HER-GNCs solution, and storing in a refrigerator at 4 ℃ for later use.
Weighing a proper amount of free maytansine derivative DM1, dissolving and diluting the free maytansine derivative DM1 into 0.5-1 mg/mL mother liquor by using acetonitrile, taking prepared gold nanocluster GNCs, mixing the GNCs and DM1 according to the mass ratio of DM1 to gold of 1: 1, stirring for 12h at room temperature, and centrifuging by using an ultrafiltration tube with the molecular weight cutoff of 10 kDa. And (3) adjusting the pH value of the prepared GNCs-DM1 to about 7.4, adding an HER2 antibody solution, uniformly mixing, lightly shaking overnight at room temperature, centrifuging and cleaning by using an ultrafiltration tube with the molecular weight cutoff of 300kDa to obtain an HER-GNCs-DM1 solution, and storing in a refrigerator at 4 ℃ for later use.
And 3mL of solution of the gold nanocluster drug-loaded targeting preparation (HER-GNCs-DM1) assembled by the polypeptide is taken, and the particle size of the solution is determined by using a Zetaplus laser particle size analyzer.
Taking a proper amount of the polypeptide-assembled gold nanocluster drug-loaded targeting preparation solution, completely wetting a copper net covered with a carbon film for 30s, then sucking off redundant liquid by using filter paper, continuously dyeing for 5min by using phosphotungstic acid, sucking off redundant liquid, drying for about 3min under light, and observing the shape and the particle size by using a transmission electron microscope.
And (3) taking 3mL of the gold nanocluster drug-loaded targeting preparation solution assembled by the polypeptide, and scanning the ultraviolet spectrum of the gold nanocluster drug-loaded targeting preparation solution by using an ultraviolet-visible spectrophotometer within the range of 400-900 nm. The results are shown in FIGS. 1, 2 and 4.
Example 3: photothermal conversion heating curve of polypeptide-assembled gold nanocluster targeting preparation
1mL of GNPs, GNCs and HER-GNCs solution was taken to have an Au concentration of 100. mu.g/mL, PBS was used as a blank, and 3W/cm was used2Continuously irradiating the 808nm near-infrared laser for 20min, measuring the temperature of the solution at intervals by using a thermal infrared imager, and drawing a temperature change curve. The results are shown in FIG. 5.
Example 4: in-vitro release curve of polypeptide-assembled gold nanocluster drug-loaded targeting preparation with/without GSH/NIR/MMP-9
The following release conditions were chosen: pH7.4 PBS; pH7.4 PBS +10mM GSH; ③ pH7.4 PBS +10mM GSH + NIR; pH7.4 PBS +10mM GSH + NIR + MMP-9; pH6.8 PBS; sixthly, the pH value is 6.8PBS and 10mM GSH; (vii) pH6.8 PBS +10mM GSH + NIR; (viii) pH6.8 PBS +10mM GSH + NIR + MMP-9. Placing 0.5mL of each of GNCs-DM1 and HER-GNCs-DM1 in a dialysis bag with molecular weight cutoff of 5kDa, fastening two ends with cotton threads, placing in a 50mL centrifugal tube, adding 10mL of corresponding release medium by taking free DM1 drug release as a control, and placing in a 37 ℃ constant temperature water bath oscillator at the rotation speed of 100 rpm/min. Taking 1mL of release medium in 0.5, 1, 2, 4, 6, 8 and 12 hours respectively, supplementing 1mL of release medium, carrying out NIR irradiation on the group to be subjected to NIR irradiation for 5min before sampling, then sampling, filtering the taken release medium through a 0.22 mu m microporous filter membrane, and measuring the release amount of DM1 by HPLC. Release curves are plotted with time as the abscissa and cumulative percent release as the ordinate. The results are shown in FIG. 6.
DM1 in the formulation was significantly released when added to the GSH release medium, and both released more than 75% at 12h at pH6.8 and pH 7.4. The release of GNCs-DM1 and HER-GNCs-DM1 was accelerated after NIR irradiation and the cumulative release at 12h was increased (xp <0.01), with GNCs-DM1 and HER-GNCs-DM1 releasing more than 80% at ph6.8 and ph 7.4. After MMP-9 was added, the final cumulative release of DM1 was also increased (p <0.05) due to the disintegration of the gold nanoclusters, which was greater than 90% in both pH6.8 and pH7.4 release media.
Example 5: matrix metalloproteinase responsiveness of gold nanoclusters
Taking a proper amount of MMP-9 in a 1.5mL centrifuge tube, adding a protein buffer solution to dilute the MMP-9 to the concentration of 100 mu g/mL, adding an APMA activating agent to enable the final concentration of the MMP-9 to be 1mM, incubating the mixture at 37 ℃ for 2h, and immediately placing the mixture in an ice tank for standby or storing the mixture at-20 ℃. Taking a proper amount of GNPs, GNCs and HER-GNCs solution, enabling the final concentration to be 1-2 nM, adding activated MMP-9 into the solution, enabling the final concentration to be 300-500 ng/mL, fully and uniformly mixing, incubating at 37 ℃, sampling for 0, 2 and 24 hours respectively, preparing a transmission electron microscope sample, and observing the form of the sample. The results are shown in FIG. 7.
With the increase of incubation time of GNCs, HER-GNCs and MMP-9, the particle size is gradually reduced, which shows that the gold nanoclusters are disintegrated into dispersed gold nanoparticles, and therefore, the gold nanoclusters have good MMP responsiveness, can realize degradation of a carrier at a tumor part, and promote tumor penetration. However, the particle size of the gold nanosphere of the unmodified responsive polypeptide is gradually increased after incubation with MMP, and electron microscope results also show that the gold nanosphere is changed into an aggregation state from dispersion, which indicates that the exposed gold nanosphere has poor stability and generates aggregation under incubation conditions.
Example 6: survival rate of SK-BR-3 and MCF-7 cells of tumor therapeutic agent free drug DM1
Collection of logarithmic SK-BR-3 and MCF-7 cells at 5000 cells/well in 96-well plates, 37 ℃, 5% CO2After 24h incubation until the cells adhere to the wall, the old culture medium is aspirated, 200 μ L of culture medium containing different concentrations of DM1 is added, a negative control group and a blank group are set simultaneously, and each group is provided with 6 duplicate wells. After 24h incubation, the well liquid is discarded, 200. mu.L of MTT (0.5mg/mL) is added into each well for further incubation for 4h, the MTT is absorbed, 150. mu.L of DMSO is added into each well, the crystals are fully dissolved by shaking for 10min, and the absorbance value of each well at 570nm is measured by a microplate reader. The results of the experiment are shown in FIG. 8. The cytotoxicity of free DM1 increased with increasing concentration, and the toxic effect of DM1 on two breast cancer cells was more obvious.
Example 7: survival rate of gold nanocluster drug-loaded targeting preparation assembled by polypeptide in SK-BR-3 and MCF-7 cells
Collection of logarithmic SK-BR-3 cells at 5000 cells/well in 96-well plates, 37 ℃ with 5% CO2After 24h incubation until the cells are attached, the old culture medium is aspirated off, 200 mu L of culture medium of GNCs-DM1 and HER-GNCs-DM1 with the same concentration (calculated as Au) is respectively added, and a negative control group and a blank group are simultaneously arranged, wherein each group is provided with 6 multiple wells. Continuing to culture for 6h, sucking out the medicinal liquid, adding 100 μ L of fresh culture medium into each well, and using power of 3W/cm2Irradiating the 808nm laser for 5min, continuously incubating for 24h after the irradiation is finished, discarding liquid in the hole, adding 200 mu L MTT (0.5mg/mL) into each hole, continuously incubating for 4h, absorbing the MTT, adding 150 mu L DMSO into each hole, shaking for 10min to fully dissolve crystals, and measuring the light absorption value of each hole at 570nm by using a microplate reader. The results of the experiment are shown in FIG. 9. The HER-GNCs-DM1 showed more obvious inhibition effect after applying near infrared laser<0.01) and the effect of the combination treatment is more pronounced with increasing concentration of gold nanoclusters, while cytotoxicity is enhanced in MCF-7 cells (. about.p)<0.01)。
Example 8: permeability of gold nanoclusters in 3D tumor sphere
The log-phase SK-BR-3 cells were harvested and diluted to 8X 10 with complete medium containing 0.24% methylcellulose4one/mL cell suspension was seeded at 25 μ L/drop onto the lid of a cell culture dish, an appropriate amount of PBS was added to the dish, and the lid with the cell drop was attached. 37 ℃ and 5% CO2After 24h incubation, the cell pellet was transferred to agarose-plated 96-well plates and culture continued to the appropriate size, with half a second change of fluid every two days. Selecting the cultured tumor spheres, administering the spheres with uniform size, compact and round shape, adding free SH-PEG-FITC, FITC-HGNPs, FITC-GNCs and FITC-HER-GNCs with the same fluorescence amount, respectively, and culturing at 37 deg.C and 5% CO2Culturing for 12 h. Simultaneously setting a group of FITC-HER-GNCs for administration for 4h, irradiating with near infrared laser with intensity of 3W/cm2Irradiating for 5min, and continuing to incubate for 12 h. Carefully aspirate the culture medium, carefully wash the tumor spheres with pre-cooled PBS 3 times, fix with 4% paraformaldehyde for 30min, rinse with PBS 3 times, transfer the tumor spheres to a confocal dish, perform Z-stack tomography using a laser confocal microscope, and take pictures. The experimental results are shown in the figureShown at 10. Free SH-PEG-FITC is difficult to permeate into the deep part of a tumor, a fluorescent signal is weak and uneven, the permeation effect of hollow gold nanoparticles with similar particle sizes on the tumor sphere is very limited, only stronger fluorescent signals are available in cells on the surface layer of the sphere, and the fluorescent signals can be detected from the surface layer to the inside of the tumor sphere by GNCs and HER-GNCs, so that the gold nanoclusters can be distributed to the deep part of the tumor more after being disintegrated under the action of MMP-9, the permeability is stronger, the fluorescent signals of the HER-GNCs modified by the antibody in the cells are more obvious, and the uptake of the tumor cells to carriers is enhanced by the antibody modification.
Example 9: gold nanocluster in vivo photoacoustic imaging
SK-BR-3 cells in logarithmic growth phase are prepared into 1 × 10 cells under aseptic condition7The cell suspension was inoculated in 0.1mL of the fourth pair of teats on the right side of each Balb/c mouse. When the tumor grows to 100-200mm3On the left and right, mice were randomly divided into 4 groups (Saline control group, HGNPs group, GNCs group, HER-GNCs group), 3 mice per group, each group of the preparation was injected from the tail vein at a dose of 10mg/kg Au, and photoacoustic imaging was performed in vivo at 0, 2, 8, and 24 hours after injection. During imaging, a mouse anesthetized by isoflurane is placed on an animal table, four limbs and the head of the mouse are fixed by a medical adhesive tape, and an ultrasonic coupling agent is coated on a tumor part to ensure that the tumor part is completely covered and has no bubbles so as to enhance the strength of photoacoustic radiography. And (3) attaching the ultrasonic probe and the coupling agent, slowly pressing down, observing the ultrasonic image, adjusting the position of the probe until the maximum section of the tumor is exposed, switching to a photoacoustic imaging mode, and taking a picture. The results of the experiment are shown in FIG. 11. The photoacoustic signals of the tumor part are gradually enhanced along with the time, and the photoacoustic signals of the HER-GNCs group are obviously stronger than those of the GNCs group and the physiological saline group, which shows that the HER-GNCs have higher tumor targeting efficiency under the dual targeting action of the active targeting and the passive targeting.
Claims (9)
1. A penetration enhanced gold nanocluster drug-loaded targeting preparation is characterized in that the gold nanocluster drug-loaded targeting preparation comprises gold nanoclusters, a cross-linking agent and a tumor treatment drug; the gold nanoclusters are obtained by assembling gold nanospheres with a cross-linking agent, and targeted tumor treatment drugs of human epidermal growth factor receptor-2 are modified on the surfaces of the gold nanoclusters;
the cross-linking agent for assembling is polypeptide with two sulfhydryl groups, and the cross-linking is gold-sulfur bond covalent bonding or electrostatic adsorption;
the double-end sulfhydryl polypeptide is double-end sulfhydryl matrix metalloproteinase sensitive polypeptide;
the amino acid sequence of the polypeptide is CRDPLGLAGDRC;
the grain diameter of the gold nanocluster is 5-15 nm, and the potential is-20 to-40 mv; the particle size of the penetration enhanced gold nano-cluster drug-loaded targeting preparation is 50-80 nm, the potential is-20 to-40 mv, and the ultraviolet maximum absorption wavelength of the penetration enhanced gold nano-cluster drug-loaded targeting preparation is 500-600 nm;
the preparation method of the permeation enhanced gold nanocluster drug-loaded targeting preparation comprises the following steps:
(1) reducing sodium borohydride to prepare gold nanospheres;
(2) crosslinking the gold nanospheres to prepare gold nanoclusters;
(3) the gold nanocluster reacts with the tumor treatment drug to prepare the gold nanocluster drug-carrying targeting preparation.
2. The penetration-enhanced gold nanocluster drug-loaded targeting preparation as claimed in claim 1, wherein the tumor therapeutic drug is a tumor therapeutic agent selected from a small molecule chemotherapeutic agent, an inorganic metal based therapeutic drug, a protein therapeutic drug or a gene therapeutic drug.
3. The osmotically enhanced gold nanocluster drug-loaded targeted formulation of claim 2, wherein the small molecule chemotherapeutic agent is selected from one or more of maytansine and its derivatives, doxorubicin hydrochloride, camptothecin, epothilone, taxane, docetaxel, daunorubicin, doxorubicin, epirubicin, mitoxantrone, idarubicin, cytarabine, erlotinib, afatinib, lapatinib, dacomitinib, gefitinib, AP32788, poetinib, ocitinib, EGF816, gertitinib, crilaitinib, tandutinib, sorafenib, midostaurin, quinatinib, pemetrexed;
the protein therapeutic drug or the tumor therapeutic agent of the gene therapeutic drug is selected from one or more of trastuzumab, AMP-224, AMP-514/MEDI-0680, astuzumab, avizumab, BGB-A317, BMS936559, Devacizumab, JTX-4014, nivolumab, pembrolizumab and SHR-1210;
the inorganic metal therapeutic drugs are gold, iron, bismuth and platinum therapeutic drugs.
4. The penetration-enhanced gold nanocluster drug-loaded targeting formulation as claimed in claim 3, wherein said small molecule chemotherapeutic agent is maytansine derivative DM 1;
the tumor therapeutic agent of the protein therapeutic drug or the gene therapeutic drug is trastuzumab;
the inorganic metal therapeutic drug carboplatin or cisplatin.
5. A preparation method of the penetration enhanced gold nanocluster drug-loaded targeting preparation as claimed in any one of claims 1 to 4, which comprises the following steps:
respectively measuring 10-25mM gold chloride acid 100-; taking 1-1.5mL of gold nanosphere, adding 1-2mL of polypeptide solution with the concentration of 1-2mg/mL, gently rotating until the mixture is uniformly mixed, wherein the polypeptide amino acid sequence is CRDPLGLAGDRC, stirring at room temperature for 12-24 hours, centrifuging by using an ultrafiltration tube with the molecular weight cutoff of 5-15kDa to obtain a gold nanocluster assembled by polypeptide, adding 1-2mg/mL of maytansine derivative DM1 solution, stirring at room temperature for 12-24 hours, adding an antibody solution dissolved in PBS, gently oscillating at room temperature for 12-24 hours, and centrifuging by using an ultrafiltration tube with the molecular weight cutoff of 300-500kDa to obtain a gold nanocluster drug-loaded targeting preparation assembled by polypeptide.
6. The method of claim 5, comprising the steps of:
(1) respectively measuring 10-25mM chloroauric acid 100-;
(2) taking 1-1.5mL of the gold nanospheres prepared in the step (1), adding 1-2mL of polypeptide solution with the concentration of 1-2mg/mL, slightly rotating until the mixture is uniformly mixed, stirring at room temperature for 12-24 hours, and centrifuging by using an ultrafiltration tube with the molecular weight cutoff of 5-15kDa to obtain polypeptide-assembled gold nanoclusters;
(3) adding 1-2mg/mL maytansine derivative DM1 solution into the polypeptide-assembled gold nanoclusters prepared in the step (2), stirring at room temperature for 12-24 hours, and centrifuging by using an ultrafiltration tube with the molecular weight cutoff of 5-15kDa to obtain a polypeptide-assembled gold nanocluster drug-carrying preparation;
(4) and (4) adding the polypeptide-assembled gold nanocluster drug-loaded preparation prepared in the step (3) into an antibody solution dissolved in PBS, slightly oscillating for 12-24 hours at room temperature, and centrifuging by using an ultrafiltration tube with the molecular weight cutoff of 300-500kDa to obtain the polypeptide-assembled gold nanocluster drug-loaded targeting preparation.
7. The use of the penetration-enhanced gold nanocluster drug-loaded targeting preparation as claimed in any one of claims 1 to 4 in the preparation of a medicament for treating malignant tumors of breast cancer, melanoma and prostate cancer.
8. The use of the penetration-enhanced gold nanocluster drug-loaded targeting preparation as claimed in any one of claims 1 to 4 in the preparation of combined radiotherapy, phototherapy and thermotherapy diagnostic agents.
9. The use of the penetration-enhanced gold nanocluster drug-loaded targeting preparation as claimed in any one of claims 1 to 4 in the preparation of diagnostic reagents for electronic computed tomography and photoacoustic imaging.
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CN107362370B (en) * | 2016-05-13 | 2022-07-26 | 国家纳米科学中心 | Method for treating pancreatic cancer based on combination of gold nanoclusters and NGF siRNA |
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Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103920889A (en) * | 2014-04-03 | 2014-07-16 | 东南大学 | Application of thiol-polyethylene glycol in preparation of water-soluble gold nano-clusters |
CN106924764A (en) * | 2017-04-24 | 2017-07-07 | 南京邮电大学 | A kind of preparation method and applications of the feature light sonic probe based on gold nano grain assembling |
WO2019190491A1 (en) * | 2018-03-27 | 2019-10-03 | National Health Research Institutes | Sub-nanometer gold sticker and methods for protecting against endotoxin-induced sepsis thereof |
CN111318718A (en) * | 2020-02-24 | 2020-06-23 | 中国科学技术大学 | Gold nano assembly with sensitization tumor radiotherapy and preparation method and application thereof |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN105770901B (en) * | 2016-03-01 | 2017-05-31 | 中国人民解放军南京军区南京总医院 | The gold nano star material of crosslinking pH response cross-film small peptides and its application |
EP3445501A4 (en) * | 2016-04-22 | 2019-11-20 | The Board of Trustees of the Leland Stanford Junior University | Synthesis of water-soluble thiolate-protected gold nanoparticles of uniform size and conjugates thereof |
-
2020
- 2020-07-24 CN CN202010723241.4A patent/CN111760024B/en active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103920889A (en) * | 2014-04-03 | 2014-07-16 | 东南大学 | Application of thiol-polyethylene glycol in preparation of water-soluble gold nano-clusters |
CN106924764A (en) * | 2017-04-24 | 2017-07-07 | 南京邮电大学 | A kind of preparation method and applications of the feature light sonic probe based on gold nano grain assembling |
WO2019190491A1 (en) * | 2018-03-27 | 2019-10-03 | National Health Research Institutes | Sub-nanometer gold sticker and methods for protecting against endotoxin-induced sepsis thereof |
CN111318718A (en) * | 2020-02-24 | 2020-06-23 | 中国科学技术大学 | Gold nano assembly with sensitization tumor radiotherapy and preparation method and application thereof |
Non-Patent Citations (5)
Title |
---|
Qishuai Feng,et al..Self-Assembly of Gold Nanoparticles Shows Microenvironment-Mediated Dynamic Switching and Enhanced Brain Tumor Targeting.《Theranostics》.2017,第7卷(第7期),第1875-1889页. * |
Self-Assembly of Gold Nanoparticles Shows Microenvironment-Mediated Dynamic Switching and Enhanced Brain Tumor Targeting;Qishuai Feng,et al.;《Theranostics》;20170410;第7卷(第7期);第1875-1889页 * |
付英杰.智能响应纳米载体的构建及其在癌症治疗中的应用研究.《中国博士学位论文全文数据库 工程科技I辑》.2019,(第02期), * |
智能响应纳米载体的构建及其在癌症治疗中的应用研究;付英杰;《中国博士学位论文全文数据库 工程科技I辑》;20190215(第02期);第B020-131页 * |
靶向肿瘤新生血管的金纳米探针用于肺癌诊治结合的研究;张璐;《中国博士学位论文全文数据库 工程科技I辑》;20200115(第01期);第B020-270页 * |
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