CN116059384A - Nanometer medicine crystal and its prepn and application - Google Patents
Nanometer medicine crystal and its prepn and application Download PDFInfo
- Publication number
- CN116059384A CN116059384A CN202211499715.7A CN202211499715A CN116059384A CN 116059384 A CN116059384 A CN 116059384A CN 202211499715 A CN202211499715 A CN 202211499715A CN 116059384 A CN116059384 A CN 116059384A
- Authority
- CN
- China
- Prior art keywords
- cpt
- hanc
- nano
- solution
- drug
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 239000003814 drug Substances 0.000 title claims abstract description 28
- 239000013078 crystal Substances 0.000 title claims abstract description 23
- 229940079593 drug Drugs 0.000 claims abstract description 26
- 238000000034 method Methods 0.000 claims abstract description 12
- 206010006187 Breast cancer Diseases 0.000 claims abstract description 9
- 208000026310 Breast neoplasm Diseases 0.000 claims abstract description 9
- 230000005764 inhibitory process Effects 0.000 claims abstract description 9
- 238000002360 preparation method Methods 0.000 claims abstract description 6
- KIUKXJAPPMFGSW-DNGZLQJQSA-N (2S,3S,4S,5R,6R)-6-[(2S,3R,4R,5S,6R)-3-Acetamido-2-[(2S,3S,4R,5R,6R)-6-[(2R,3R,4R,5S,6R)-3-acetamido-2,5-dihydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-2-carboxy-4,5-dihydroxyoxan-3-yl]oxy-5-hydroxy-6-(hydroxymethyl)oxan-4-yl]oxy-3,4,5-trihydroxyoxane-2-carboxylic acid Chemical compound CC(=O)N[C@H]1[C@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@H](O3)C(O)=O)O)[C@H](O)[C@@H](CO)O2)NC(C)=O)[C@@H](C(O)=O)O1 KIUKXJAPPMFGSW-DNGZLQJQSA-N 0.000 claims description 35
- 229920002674 hyaluronan Polymers 0.000 claims description 35
- 229960003160 hyaluronic acid Drugs 0.000 claims description 35
- 239000010931 gold Substances 0.000 claims description 22
- 239000002105 nanoparticle Substances 0.000 claims description 16
- 229910021642 ultra pure water Inorganic materials 0.000 claims description 11
- 239000012498 ultrapure water Substances 0.000 claims description 11
- 238000002156 mixing Methods 0.000 claims description 10
- 239000002245 particle Substances 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 9
- 238000010438 heat treatment Methods 0.000 claims description 8
- 101710134784 Agnoprotein Proteins 0.000 claims description 4
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
- 230000001105 regulatory effect Effects 0.000 claims description 4
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 claims description 3
- 229910052737 gold Inorganic materials 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- 238000003860 storage Methods 0.000 claims description 3
- FOIXSVOLVBLSDH-UHFFFAOYSA-N Silver ion Chemical compound [Ag+] FOIXSVOLVBLSDH-UHFFFAOYSA-N 0.000 claims description 2
- 230000002776 aggregation Effects 0.000 claims description 2
- 238000004220 aggregation Methods 0.000 claims description 2
- 239000006185 dispersion Substances 0.000 claims description 2
- 230000001678 irradiating effect Effects 0.000 claims description 2
- 239000002159 nanocrystal Substances 0.000 claims description 2
- -1 Ag/HA Chemical compound 0.000 claims 1
- 230000001093 anti-cancer Effects 0.000 abstract description 6
- 230000002401 inhibitory effect Effects 0.000 abstract description 4
- 229940044683 chemotherapy drug Drugs 0.000 abstract description 3
- 230000007246 mechanism Effects 0.000 abstract description 3
- 239000002086 nanomaterial Substances 0.000 abstract description 3
- 230000009471 action Effects 0.000 abstract description 2
- 239000002246 antineoplastic agent Substances 0.000 abstract description 2
- 239000002923 metal particle Substances 0.000 abstract description 2
- 230000003287 optical effect Effects 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 25
- 206010028980 Neoplasm Diseases 0.000 description 21
- 238000011282 treatment Methods 0.000 description 13
- 210000000130 stem cell Anatomy 0.000 description 7
- 238000006243 chemical reaction Methods 0.000 description 6
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 206010027476 Metastases Diseases 0.000 description 3
- 201000011510 cancer Diseases 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 230000009401 metastasis Effects 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 206010061309 Neoplasm progression Diseases 0.000 description 2
- 230000003698 anagen phase Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000002296 dynamic light scattering Methods 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000013532 laser treatment Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000007626 photothermal therapy Methods 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 230000005751 tumor progression Effects 0.000 description 2
- YLNDNABNWASMFD-UHFFFAOYSA-N 4-[(1,3-dimethylimidazol-1-ium-2-yl)diazenyl]-n,n-dimethylaniline Chemical compound C1=CC(N(C)C)=CC=C1N=NC1=[N+](C)C=CN1C YLNDNABNWASMFD-UHFFFAOYSA-N 0.000 description 1
- 102100032912 CD44 antigen Human genes 0.000 description 1
- 230000005778 DNA damage Effects 0.000 description 1
- 231100000277 DNA damage Toxicity 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 101000868273 Homo sapiens CD44 antigen Proteins 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- FZWLAAWBMGSTSO-UHFFFAOYSA-N Thiazole Chemical compound C1=CSC=N1 FZWLAAWBMGSTSO-UHFFFAOYSA-N 0.000 description 1
- 238000004115 adherent culture Methods 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 229910021645 metal ion Inorganic materials 0.000 description 1
- 239000002082 metal nanoparticle Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 229910052709 silver Inorganic materials 0.000 description 1
- 239000004332 silver Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/242—Gold; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K33/00—Medicinal preparations containing inorganic active ingredients
- A61K33/24—Heavy metals; Compounds thereof
- A61K33/38—Silver; Compounds thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Inorganic Chemistry (AREA)
- Epidemiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicinal Preparation (AREA)
Abstract
The invention belongs to the field of biomedical nano materials, and discloses a nano drug crystal and a preparation method and application thereof. The nano-drug crystal component comprises Au/HA, ag/HA and CPT/HANC, and can be applied to the inhibition of conventional breast cancer cells and BCSCs. Based on the diversity and complementarity of AuNPs and AgNPs for inhibiting the action mechanism of the CSCs, the Au-Ag@CPT/HANC prepared by the method can integrate the characteristics of two metal particles, has unique optical and biological characteristics, and is assisted with the anticancer characteristics of chemotherapeutic drugs, so that the aim of inhibiting the residual of the CSCs by multiple targets is fulfilled.
Description
Technical Field
The invention belongs to the field of biomedical nanomaterials, and particularly relates to a bimetallic nanoparticle modified nano-drug crystal (Au-Ag@CPT/HANC) for resisting Breast Cancer Stem Cells (BCSCs), and a preparation method and application thereof.
Background
Breast cancer is one of the most common cancers in women. Despite the high survival rate after treatment, breast cancer patients remain at high risk of tumor recurrence and metastasis. The reason for this is that while chemotherapy drugs can kill most of the normal tumor cells in the rapidly proliferating stage in tumor tissue, reducing the volume of tumor tissue mass, it is difficult to eliminate a relatively low content of a subset of cells known as tumor stem cells (CSCs). CSCs are located at the top of the cell hierarchy and are very low in tumor content, but are probably key cells for tumor progression, they have characteristics similar to those of normal stem cells, such as self-renewal, multidirectional differentiation, high expression of certain stem-related genes, and the like, are closely related to tumor progression, drug resistance, recurrence, metastasis, and the like, and are root causes of cancer recurrence and metastasis. And the residual phenomenon of the CSCs, which is caused by various mechanisms, can be inhibited by multiple targets only by developing a multi-component combined therapeutic strategy of the CSCs.
In recent years, nanoparticles have been found to be an important choice for the treatment of CSCs resistance. There is more evidence that nanoparticle-mediated photothermal therapy can stimulate a thermophysiologic response to kill cancer cells, as a non-invasive treatment, leading to DNA damage and protein denaturation in tumors, etc. leading to apoptosis.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention provides a nano-drug crystal, a preparation method and application thereof, and the nano-drug crystal has high biocompatibility and a certain photo-thermal effect and can effectively inhibit the formation of tumor balls.
The above object of the present invention is achieved by the following technical solutions: a method of preparing a nanocrystal comprising the steps of:
ha solution and HAuCl respectively 4 、AgNO 3 Mixing the solutions, regulating the pH of the solutions after ultra-pure water is subjected to constant volume, heating and stirring in a dark place to obtain Hyaluronic Acid (HA) -coated single gold nano particles (Au/HA) and hyaluronic acid-coated single silver nano particles (Ag/HA), and then respectively centrifugally collecting the Au/HA and the Ag/HA and re-suspending for storage.
2. Adding 400 mu L of 0.01-10 mg/mL CPT solution into HA solution with the concentration of 0.001-1 mg/mL, carrying out magnetic stirring in a dark place after ultrasonic crushing, and filtering by a carbonate membrane (with the pore diameter of 10-4000 nm) to obtain CPT/HANC.
3. And (2) uniformly mixing the Au/HA and Ag/HA solutions obtained in the step (1) in a certain molar ratio, adding the ultrapure water to the constant volume, adding the CPT/HANC obtained in the step (2), and magnetically stirring in a dark place to obtain the nano-drug crystal modified by the bimetallic nano-particles (Au-Ag@CPT/HANC).
4. And (3) placing the nano-drug crystal Au-Ag@CPT/HANC obtained in the step (3) in 808nm near infrared laser, and irradiating for 0-2 min under 2-4W power.
Further, the HA solution and the HAuCl in the step 1 4 The mixing mole ratio of the solution is 1:1-1:4, the pH is regulated to 7.5-9.5, and the light-shielding heating temperature is 65-75 ℃.
Further, in the step 1, the HA solution and AgNO 3 The mixing mole ratio of the solution is 1:10-1:3, the pH is adjusted to 8.5-11.0, and the temperature of the light-shielding heating is 65-75 ℃.
Further, in the step 3, au/HA: ag/HA: the molar ratio of CPT/HANC is 10:10:1-1:4:3.
The invention also aims to protect the nano-drug crystal prepared by the nano-drug crystal preparation method, wherein the particle size of Au/HA is 20+/-20 nm-500+/-100 nm, the particle size of Ag/HA is 20+/-20 nm-500+/-100 nm, the particle size of CPT/HANC is 50+/-20 nm-3000+/-100 nm, the particle size of Au-Ag@CPT/HANC is 100+/-50 nm-4000+/-100 nm, and the Au/HA and the Ag/HA are respectively interspersed around the rod-shaped CPT/HANC in a similar spherical shape in an aggregation sample or a dispersion sample.
It is a further object of the present invention to protect the above-mentioned nanocrystalline drug crystals from the effects of other agents, including the inhibition of conventional breast cancer cells and BCSCs.
Under 808nm near infrared laser, the Au-Ag@CPT/HANC suspension can be increased in temperature increment along with the increase of laser power and irradiation time, and the temperature increment can reach 40-50 ℃ within 0-2 min under the power condition of 2-4W when the CPT concentration is 1-10 mug/mL.
Compared with the prior art, the invention has the beneficial effects that: based on the diversity and complementarity of AuNPs and AgNPs for inhibiting the action mechanism of CSCs, au is prepared in the applicationThe Ag@CPT/HANC can integrate the characteristics of two metal particles, has unique optical and biological characteristics, and is matched with the anticancer characteristics of chemotherapeutic drugs to realize the aim of inhibiting the residual of the CSCs by multiple targets. Meanwhile, the balance point of high anticancer activity and low physiological toxicity can be determined by effectively adjusting the proportion of gold and silver in the nano drug crystal; the application uses the natural linear polysaccharide HA as the biological reducing agent to prepare the bimetallic nanoparticle simply, conveniently and environmentally, namely, the reducing group of the HA is utilized to induce metal ions (Au 3+ 、Ag + ) Is to chemically reduce and fix the nascent metal nanoparticles (Au 0 、Ag 0 ). In addition, HA can be used as a targeting molecule to be modified on the surface of a nano drug delivery system, so that breast cancer cells expressing CD44 at high level are specifically targeted, the compatibility of nano materials is improved, and the blood circulation period of the drug is prolonged. The Au-Ag@CPT/HANC and the Au/HA prepared by the method have high photo-thermal conversion efficiency and have remarkable inhibition effect on tumor balls.
Drawings
The invention will be further described with reference to the drawings and the detailed description
FIG. 1 is a Scanning Electron Microscope (SEM) image (a) and an element analysis image (EDX) (b) of Au-Ag@CPT/HANC prepared by the method;
FIG. 2 is a graph showing the experimental result of cytotoxicity of MDA-MB-231 of Au-Ag@CPT/HANC prepared by the method;
FIG. 3 is a bar chart showing the inhibition of BCSCs by Au-Ag@CPT/HANC prepared by the method of the invention;
FIG. 4 is a schematic diagram showing the inhibition of the BCSCs by Au-Ag@CPT/HANC prepared by the method.
Detailed Description
The present invention is described in detail below by way of specific examples, but the scope of the present invention is not limited thereto. Unless otherwise specified, the experimental methods used in the present invention are all conventional methods, and all experimental equipment, materials, reagents, etc. used can be obtained from commercial sources.
Example 1
The preparation method of the nano-drug crystal modified by the bimetallic nano-particles for resisting breast cancer stem cells comprises the following specific steps:
s1, respectively removing 100 mu LHAuCl by using a microsampler 4 Solution and 1mLHA solution, to give HA: au in the reaction solution 3 + And (3) adjusting the pH value of the reaction solution to 7.5-9.5 after the ultrapure water is subjected to constant volume at a ratio of 1:2, and heating and stirring in a dark place to obtain Au/HA. The Au/HA obtained is along with Au 3+ The color of the solution gradually deepens from light purple to ruby red due to the increase of the content. The Au/HA was then obtained by centrifugation, followed by re-suspension in ultra pure water and storage in the dark.
S2, respectively removing 50 mu LAgNO by using a microsampler 3 Solution and 1mLHA solution, to make HA: ag in the reaction solution + And (3) adjusting the pH value of the reaction solution to 8.5-11.0 after the ultrapure water is subjected to constant volume at a ratio of 1:1, and heating and stirring in a dark place to obtain Ag/HA. The obtained Ag/HA follows Ag + The color of the solution gradually deepens from light brown to dark brown due to the increase of the content. Then, the mixture was centrifuged to obtain Ag/HA, which was then resuspended in ultra-pure water and stored in a dark place.
S3, removing ultrapure water with the pH value of 4 by using a microsampler, adding 100 mu LHA solution, uniformly mixing, dropwise adding 400 mu LCPT solution, magnetically stirring for 3 hours at room temperature in a dark state after ultrasonic crushing, and filtering by using a carbonate membrane to obtain CPT/HANC.
S4, respectively removing 1mLAu/HA and 4mL of Ag/HA solution obtained in the steps S1 and S2 by using a microsampler; and (3) enabling Au to Ag in the reaction solution to be 1:2, uniformly mixing, then, carrying out ultrapure water constant volume, and then, adding CPT/HANC obtained in the step (S3). Magnetically stirring at room temperature in a dark place to obtain Au-Ag@CPT/HANC, filtering by a carbonate film, and then re-suspending with ultrapure water and preserving in a dark place. Subsequently, physicochemical property analysis is carried out on the nano particles provided by the invention, and an SEM (scanning electron microscope) and an EDX (electron microscope) diagram of Au-Ag@CPT/HANC (hybrid electro-magnetic resonance) at the optimal volume ratio of 1:2 provided by the invention are shown in figure 1. The SEM result is consistent with the analysis result of Dynamic Light Scattering (DLS), the particle size is 100+/-50 nm-4000+/-100 nm, and the distribution is uniform.
S5, in vitro anticancer activity assay (MTT method): the invention selects MDA-MB-231 breast cancer cell line to explore the in vitro anticancer activity of Au-Ag@CPT/HANC. Taking 1×10 cells in logarithmic growth phase 4 Inoculating the cells/wells into 96-well cell culture plate, overnight culturing, adding different nanoparticle forms (negative control, CPT/HANC, CPT/HANC+L, etc.),Au-Ag@CPT/HANC, au-Ag@CPT/HANC+L) for 24 hours; the negative control was cells without any treatment. Wherein CPT/HANC+ L, au-Ag@CPT/HANC+L is a drug treatment for 24 hours, and 808nm laser treatment (3.5W/cm) 2 1 min). After the drug treatment is finished, 20 mu L of 5mg/mL thiazole blue solution is added into each hole, the supernatant is sucked and removed after incubation is continued for 4 hours at 37 ℃, then 100 mu L/hole of dimethyl sulfoxide (DMSO) is added, and the OD value of a sample is measured at the wavelength of 570nm and 650nm by using an enzyme-labeled instrument, so that the survival rate of the cells is calculated.
FIG. 2 shows cytotoxicity of MDA-MB-231 cells after treatment with different pharmaceutical dosage forms. From the analysis, CPT/HANC showed less damage to cells, and nearly 70% of cells survived even when CPT concentration reached 25. Mu.g/mL. Compared with the control group, the cell survival rate of the Au-Ag@CPT/HANC treatment group is reduced to 40%, and the most remarkable is that the Au-Ag@CPT/HANC+L treatment group has the greatest killing power on cells, and only 18% of cells survive. In addition, the nano-drug crystal modified by the calculated bimetallic nano-particles has the strongest cytotoxicity combined with near infrared laser irradiation treatment group and IC 50 Is 2-3 mug/mL, which is obviously lower than the other groups. Thus, the combined use of the bimetallic nanoparticle modified nano-drug crystal and photothermal therapy provided by the invention can be initially proved to obviously improve the anticancer activity of CPT.
S6, tumor sphere inhibition: tumor sphere formation is related to the proportion of CSCs in cancer cells, so the inhibition of CSCs by Au-ag@cpt/HANC can be assessed by the number and volume of tumor spheres formed. Taking MDA-MB-231 cells in logarithmic growth phase, passaging by conventional method, and culturing at 1×10 6 The individual cells/well were inoculated into 6-well cell culture plates, after overnight adherent culture, the culture solution was removed, and different nanoparticle formulations (negative control, CPT/HANC, CPT/HANC+ L, au-Ag@CPT/HANC, au-Ag@CPT/HA NC+L, CPT concentration of 10. Mu.g/mL) were added for incubation for 24h. Wherein CPT/HANC+ L, au-Ag@CPT/HANC+L is a drug treatment for 24 hours, and 808nm laser treatment (3.5W/cm) 2 1 min), and incubating again for 3h. The cells were then digested with 25% pancreatin and counted, and the treated cells were counted at 1X 10 4 Inoculating the cells/wells into ultra-low adhesion 24-well plate, culturing in stem cell culture solution for 10 days in suspension, and displaying in inverted stateCounting tumor balls under micro-mirror>50 μm defined as stem cells).
Fig. 3 and 4 are schematic diagrams showing the number of tumor nodules and the inhibition result of tumor nodules after treating cells with different nanoparticles. Compared with CPT/HANC and CPT/HANC+L treatment groups, the Au-Ag@CPT/HANC group remarkably reduces the tumor sphere forming capacity (about 75%), and the Au-Ag@CPT/HANC+L group is difficult to aggregate into spheres after treatment and is mostly dispersed single cells. Therefore, the nano-drug crystal modified by the bimetallic nano-particles can treat drug-resistant tumors based on the survival of tumor stem cells, and has remarkable effect.
The above-described embodiments are only preferred embodiments of the invention, and not all embodiments of the invention are possible. Any obvious modifications thereof, which would be apparent to those skilled in the art without departing from the principles and spirit of the present invention, should be considered to be included within the scope of the appended claims.
Claims (7)
1. A method for preparing a nanocrystal comprising the steps of:
ha solutions with HAuCl, respectively 4 、AgNO 3 Mixing the solutions, respectively regulating the pH of the solutions after the ultrapure water is subjected to constant volume, heating and stirring in a dark place to respectively obtain hyaluronic acid coated single gold nano particles, namely Au/HA and hyaluronic acid coated single silver nano particles, namely Ag/HA, and then respectively centrifugally collecting the Au/HA and the Ag/HA and re-suspending for storage;
B. adding 400 mu L of 0.01-10 mg/mL CPT solution into HA solution with the concentration of 0.001-1 mg/mL, carrying out ultrasonic crushing, magnetically stirring in a dark place, and filtering by using a carbonate membrane with the aperture range of 10-4000 nm to obtain CPT/HANC;
C. taking the Au/HA and Ag/HA solution obtained in the step A, uniformly mixing, then adding the ultrapure water to fix the volume, adding the CPT/HANC obtained in the step B, and magnetically stirring in a dark place to obtain a nano drug crystal modified by the bimetallic nanoparticles, namely Au-Ag@CPT/HANC;
D. and C, placing the nano-drug crystal Au-Ag@CPT/HANC obtained in the step C in 808nm near infrared laser, and irradiating for 0-2 min under the power of 2-4W.
2. The method of claim 1, wherein HA solution and HAuCl are mixed in step a 4 The mixing mole ratio of the solution is 1:1-1:4, the pH is regulated to 7.5-9.5, and the light-shielding heating temperature is 65-75 ℃.
3. The method of claim 1, wherein HA solution and AgNO in step a are mixed with each other 3 The mixing mole ratio of the solution is 1:10-1:3, the pH is adjusted to 8.5-11.0, and the temperature of the light-shielding heating is 65-75 ℃.
4. The method of preparing nano-drug crystals according to claim 1, wherein Au/HA in step C: ag/HA: the molar ratio of CPT/HANC is 10:10:1-1:4:3.
5. The nano-drug crystal prepared by the preparation method of claim 1, wherein the particle size of Au/HA is 20+/-20 nm-500+/-100 nm, the particle size of Ag/HA is 20+/-20 nm-500+/-100 nm, the particle size of CPT/HANC is 50+/-20 nm-3000+/-100 nm, the particle size of Au-Ag@CPT/HANC is 100+/-50 nm-4000+/-100 nm, and the Au/HA and the Ag/HA are respectively interspersed around the rod-shaped CPT/HANC in a quasi-spherical aggregation sample or dispersion sample.
6. Use of a nano-drug crystal according to claim 5.
7. The use of nano-drug crystals according to claim 6, characterized by application in the inhibition of conventional breast cancer cells and BCSCs.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211499715.7A CN116059384A (en) | 2022-11-28 | 2022-11-28 | Nanometer medicine crystal and its prepn and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211499715.7A CN116059384A (en) | 2022-11-28 | 2022-11-28 | Nanometer medicine crystal and its prepn and application |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116059384A true CN116059384A (en) | 2023-05-05 |
Family
ID=86169026
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211499715.7A Pending CN116059384A (en) | 2022-11-28 | 2022-11-28 | Nanometer medicine crystal and its prepn and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116059384A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110278497A1 (en) * | 2010-05-11 | 2011-11-17 | China Medical University | Nanometal dispersion and method for preparing the same |
| CN108210925A (en) * | 2018-04-03 | 2018-06-29 | 国家纳米科学中心 | A kind of Nano medication and its preparation method and application |
| CN110652518A (en) * | 2019-05-05 | 2020-01-07 | 海南亚洲制药股份有限公司 | Targeting type nano drug-loading system and preparation method thereof |
| CN111437388A (en) * | 2019-01-15 | 2020-07-24 | 上海大学 | Gold-silver alloy nano material, preparation method and application thereof |
| CN115282271A (en) * | 2022-06-21 | 2022-11-04 | 大连工业大学 | Gold/silver alloy nanoparticle for resisting breast cancer stem cells and preparation method and application thereof |
-
2022
- 2022-11-28 CN CN202211499715.7A patent/CN116059384A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110278497A1 (en) * | 2010-05-11 | 2011-11-17 | China Medical University | Nanometal dispersion and method for preparing the same |
| CN108210925A (en) * | 2018-04-03 | 2018-06-29 | 国家纳米科学中心 | A kind of Nano medication and its preparation method and application |
| CN111437388A (en) * | 2019-01-15 | 2020-07-24 | 上海大学 | Gold-silver alloy nano material, preparation method and application thereof |
| CN110652518A (en) * | 2019-05-05 | 2020-01-07 | 海南亚洲制药股份有限公司 | Targeting type nano drug-loading system and preparation method thereof |
| CN115282271A (en) * | 2022-06-21 | 2022-11-04 | 大连工业大学 | Gold/silver alloy nanoparticle for resisting breast cancer stem cells and preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| JIHUI WANG ET AL.: ""Hyaluronic Acid-Coated Camptothecin Nanocrystals for Targeted Drug Delivery to Enhance Anticancer Efficacy"", 《MOL. PHARMACEUTICS》, vol. 2020, no. 17, 21 May 2020 (2020-05-21), pages 2411 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | Se@ SiO 2–FA–CuS nanocomposites for targeted delivery of DOX and nano selenium in synergistic combination of chemo-photothermal therapy | |
| Zhang et al. | Tumor environment responsive degradable CuS@ mSiO2@ MnO2/DOX for MRI guided synergistic chemo-photothermal therapy and chemodynamic therapy | |
| Ni et al. | Gold nanorod embedded large-pore mesoporous organosilica nanospheres for gene and photothermal cooperative therapy of triple negative breast cancer | |
| CN113289030B (en) | Preparation method of targeting long-circulating nano-drug carrier for photo-thermal synergistic chemotherapy | |
| Xu et al. | Hollow mesoporous manganese oxides: application in cancer diagnosis and therapy | |
| Song et al. | Fabrication of the biomimetic DOX/Au@ Pt nanoparticles hybrid nanostructures for the combinational chemo/photothermal cancer therapy | |
| CN112641946A (en) | Polydopamine-coated gold nano-composite, preparation method thereof and application thereof in multi-modal tumor diagnosis and treatment | |
| Al-Kinani et al. | Study the effect of laser wavelength on polymeric metallic nanocarrier synthesis for curcumin delivery in prostate cancer therapy: in vitro study | |
| Dai et al. | SiO2-coated magnetic nano-Fe3O4 photosensitizer for synergistic tumour-targeted chemo-photothermal therapy | |
| Zhang et al. | Noble metal nanoparticles: Synthesis, and biomedical implementations | |
| Zhang et al. | Research progress in the synthesis and biological application of quantum dots | |
| CN112843250A (en) | Nanometer medicinal preparation for treating tumor with iron death and gas synergistic effect, and its preparation method | |
| CN107281494B (en) | Preparation method and application of graphene oxide-protamine/sodium alginate compound | |
| Chen et al. | Tumor-specific nanomedicine via sequential catalytic reactions for accurate tumor therapy | |
| Han et al. | BSA-stabilized mesoporous organosilica nanoparticles reversed chemotherapy resistance of anaplastic thyroid cancer by increasing drug uptake and reducing cellular efflux | |
| Kang et al. | Flexible human serum albumin nanocapsules to enhance drug delivery and cellular uptake for photodynamic/chemo cancer therapy | |
| CN113350524B (en) | Iron-blocking magnetic nano-drug responding to tumor microenvironment as well as preparation method and application thereof | |
| He et al. | Polymyxin E biomineralized and doxorubicin-loaded gold nanoflowers nanodrug for chemo-photothermal therapy | |
| CN112791061B (en) | Preparation method of multi-stage bionic nano-drug carrier with targeting long circulation | |
| CN109224063A (en) | The nano combined carrier and its preparation and application of dual load peptides and chemotherapeutics | |
| Wang et al. | Progress in cancer therapy with functionalized Fe3O4 nanomaterials | |
| CN104825488A (en) | Loading arsenical agent as well as preparation method and application thereof | |
| JP2019043798A (en) | Dispersible calcium phosphate nanoparticle | |
| CN109550050B (en) | Melanin-loaded molybdenum dioxide drug-loaded compound and preparation and application thereof | |
| Guo et al. | The facile formation of hierarchical mesoporous silica nanocarriers for tumor-selective multimodal theranostics |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |