CN114917191B - Preparation method and application of vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS - Google Patents
Preparation method and application of vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS Download PDFInfo
- Publication number
- CN114917191B CN114917191B CN202210370030.6A CN202210370030A CN114917191B CN 114917191 B CN114917191 B CN 114917191B CN 202210370030 A CN202210370030 A CN 202210370030A CN 114917191 B CN114917191 B CN 114917191B
- Authority
- CN
- China
- Prior art keywords
- vitamin
- vfns
- optic
- photo
- acousto
- 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.)
- Active
Links
Classifications
-
- 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
- A61K41/0028—Disruption, e.g. by heat or ultrasounds, sonophysical or sonochemical activation, e.g. thermosensitive or heat-sensitive liposomes, disruption of calculi with a medicinal preparation and ultrasounds
- A61K41/0033—Sonodynamic cancer therapy with sonochemically active agents or sonosensitizers, having their cytotoxic effects enhanced through application of ultrasounds
-
- 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
- A61K33/26—Iron; Compounds thereof
-
- 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
-
- 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
- A61K47/50—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/54—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
- A61K47/55—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 the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y10/00—Nanotechnology for information processing, storage or transmission, e.g. quantum computing or single electron logic
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y15/00—Nanotechnology for interacting, sensing or actuating, e.g. quantum dots as markers in protein assays or molecular motors
-
- B—PERFORMING OPERATIONS; TRANSPORTING
- B82—NANOTECHNOLOGY
- B82Y—SPECIFIC USES OR APPLICATIONS OF NANOSTRUCTURES; MEASUREMENT OR ANALYSIS OF NANOSTRUCTURES; MANUFACTURE OR TREATMENT OF NANOSTRUCTURES
- B82Y40/00—Manufacture or treatment of nanostructures
-
- 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
-
- 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)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Nanotechnology (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Crystallography & Structural Chemistry (AREA)
- Molecular Biology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physics & Mathematics (AREA)
- Theoretical Computer Science (AREA)
- Oncology (AREA)
- Inorganic Chemistry (AREA)
- Mathematical Physics (AREA)
- General Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Manufacturing & Machinery (AREA)
- Organic Chemistry (AREA)
- Condensed Matter Physics & Semiconductors (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Medical Informatics (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The invention relates to the technical field of nano materials, in particular to a preparation method and application of vitamin B2-based acousto-optic (photo) sensitive nano particles VFNS. The nanometer medicine uses vitamin B2 as ligand, and iron ion (Fe) 3+ ) For connecting anchor points, the nano-drug is assembled through coordination driving, the operation is simple, the cost is low, no redundant carrier is used for increasing the metabolism burden of the organism, and the method comprises the following steps: (1) Precisely weighing a certain amount of FeCl 3 ·6H 2 O; (2) Precisely weighing a certain amount of vitamin B2, and ultrasonically centrifuging and dissolving the vitamin B2 by using pure water; (3) Precisely measuring vitamin B2 solution into ampoule bottle, and slowly adding FeCl under stirring of magnetic stirrer 3 ·6H 2 And O, stirring at room temperature for a period of time until the system has obvious Tyndall effect. The vitamin B2-based acousto-optic (photo) sensitive nano particles prepared by the invention have good stability and high biological safety, can generate Reactive Oxygen Species (ROS) under the irradiation of ultrasonic and blue light, and have the effects of acousto-optic power treatment and photo-optic power treatment.
Description
Technical Field
The invention relates to the technical field of nano materials, in particular to a preparation method and application of vitamin B2-based acousto-optic (photo) sensitive nano particles VFNS.
Background
Cancer has become one of the major diseases worldwide threatening human health. Although traditional chemotherapeutics achieve anticancer effects to a certain extent, due to the fact that the chemotherapeutics are rapidly cleared when in human blood circulation, targeting to tumors is poor, the traditional chemotherapeutics are limited in curative effect and have serious toxic and side effects. For many years, development of a novel drug delivery system with stable in vivo metabolic properties, good targeting and high anticancer efficiency has been the focus of anti-tumor research. Vitamin B2 is a vitamin B group, has high biological safety, participates in vivo biological oxidation and energy metabolism, and promotes development and cell regeneration. Meanwhile, the vitamin B2 has photosensitive property, can generate Reactive Oxygen Species (ROS) under the irradiation of monochromatic light, and can be used for high-efficiency tumor photodynamic therapy (PDT). PDT is a non-invasive anti-tumor therapy that uses a combination of photosensitizers, tissue oxygen, and light to produce cytotoxic reactive oxygen species to achieve tumor cell killing effects. However, photodynamic effects are limited by the depth of penetration of the photoptissue, and phototoxicity of photosensitizers to the skin limits their clinical application. For this reason, there is an urgent need to develop new methods for enhancing the antitumor therapeutic effect of vitamin B2, starting from new therapeutic approaches.
As an alternative therapy to PDT, sonodynamic therapy (SDT) may use both sonosensitizers and low intensity Ultrasound (US) to kill malignant tumors. SDT has many advantages over PDT, including high accuracy, deeper tissue penetration, good patient compliance, and fewer side effects. Ultrasound can penetrate deep tissues and focus on tumor areas, thereby activating sonosensitizers, providing the possibility of non-invasive targeted eradication of solid tumors. Ultrasound has been widely used clinically for disease diagnosis and treatment due to its noninvasive nature, extremely low energy attenuation, and high biological tissue penetrating power. However, the low quantum yield of hypoxic Tumor Microenvironment (TME) and sonosensitizers limits the therapeutic efficiency of SDT; in addition, high levels of Glutathione (GSH) in TME lead to increased resistance of cells to ROS, thereby inhibiting tumor cell apoptosis.
The use of other non-apoptotic cell death modalities opens up new therapeutic approaches for eradicating cancer cells and limiting survival of anti-apoptotic cells. In recent years, iron death has attracted attention from many researchers in the tumor field. Iron death is a novel form of programmed cell death driven by iron-dependent phospholipid peroxidation, as distinguished from apoptosis, cell necrosis, autophagy. Iron death is characterized by depletion of intracellular reduced GSH, deactivation of glutathione peroxidase 4 (GPX 4), accumulation of Lipid Peroxides (LPO), and eventually a decrease in cell volume and an increase in mitochondrial membrane density. Iron death can play a therapeutic role in a variety of anti-apoptotic tumors, such as gastric cancer, pancreatic cancer, colorectal cancer, hepatocellular carcinoma, and the like. Therefore, inducing iron death of tumor cells opens a new way for cancer treatment.
With the continuous development of various anti-tumor strategies, the combination therapy of two or more medicaments can aim at different medicinal mechanisms and different cancer passages, and the advantages of various anti-cancer methods can be hopefully utilized, so that the defect is overcome, and a better anti-tumor treatment effect is realized. However, there is still a lack of reliable solutions.
Disclosure of Invention
The invention aims to provide a preparation method and application of vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS, and aims to obtain vitamin B2 and Fe by a simple, green and safe method 3+ Coordinated nanoparticles and their application in sonodynamic (photodynamic) and iron death treatment of tumors to solve the problems set forth in the background art above.
In order to achieve the above purpose, the present invention provides the following technical solutions:
a process for preparing the sound (light) sensitive nano-particles VFNS based on vitamin B2 includes such steps as preparing the nano-particles VFNS from the polyhydroxy structure of vitamin B2 and Fe 3+ Coordination composition, the reaction equation is shown as follows:
preferably, the nanoparticle VFNS specifically comprises the following steps:
step one: precisely weighing FeCl 3 ·6H 2 O；
Step two: precisely weighing vitamin B2, and ultrasonically centrifuging and dissolving with pure water to obtain vitamin B2 water solution;
step three: essenceWeighing the vitamin B2 aqueous solution in a dense amount into an ampoule bottle, and slowly adding the FeCl which is precisely weighed under the stirring of a magnetic stirrer 3 ·6H 2 O, continuing stirring for a period of time until the system has obvious Tyndall effect; and (3) dialyzing or ultrafiltering and concentrating by pure water, collecting nano-liquid and freeze-drying to obtain the required nano-particles, wherein the particle size of the nano-particles is 50-100 nm.
Preferably, in the third step, the concentration of the vitamin B2 aqueous solution is 1-10 mg/100mL, feCl 3 ·6H 2 The amount of the O substance is 0.185mmol to 0.370mmol; by adjusting FeCl 3 ·6H 2 The amount of O to achieve the effect of FeCl 3 ·6H 2 Control of O/vitamin B2 nanoparticle size.
Preferably, in the second step, the rotational speed of the centrifuge is 10000rpm and the centrifugation time is 10 minutes.
Preferably, in the third step, the reaction temperature is room temperature, the reaction time is 30-60 min, and the rotation speed of the stirrer is 1000rpm.
Preferably, in step three, the dialysis time is 12 to 24 hours.
Preferably, in the third step, or precisely weighing the vitamin B2 aqueous solution into an ampoule bottle, slowly adding the precisely weighed FeCl under the stirring of a magnetic stirrer 3 ·6H 2 O, continuing stirring for a period of time until the system has obvious Tyndall effect; and (3) using an ultrasonic cell grinder to carry out ultrasonic treatment under certain power and time, collecting nano liquid and freeze-drying to obtain the required nano particles. Wherein the ultrasonic power is 50-200W, the ultrasonic time is 5-10s, the ultrasonic gap time is 1-5s, and the total duration is 1-10min.
The invention also provides an application of the vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS in resisting tumors.
The invention also provides an application of the vitamin B2-based acousto-optic (photo) sensitive nano particle VFNS in acousto-optic (photo) power and iron death.
Compared with the prior art, the invention has the beneficial effects that:
1. the vitamin B2-based acousto-optic (photo) sensitive nano particles obtained by the invention have uniform size, good dispersibility, simple and quick synthesis method and convenient production.
2. The vitamin B2-based acousto-optic (photo) sensitive nano particles obtained by the invention have good biological safety and high stability, can generate a large amount of active oxygen under the irradiation of acousto-optic (photo), and can be used for acousto-optic (photo) dynamic treatment of tumors.
3. The vitamin B2-based acousto-optic (photo) sensitive nano particles prepared by the invention can be prepared by Fe 3+ Reducing reaction with GSH, reducing GSH content in tumor cells, and increasing lipid peroxide content in tumor cells by inhibiting GPX4 passage, thereby achieving the effect of treating tumor by iron death.
4. The vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS with high biological safety/stability is synthesized by a one-step method, and the obtained VFNS has the characteristics of high quantum yield of active oxygen, excellent biocompatibility and dispersibility, and great potential in acousto-optic (photo) dynamic treatment and iron death treatment of tumors.
Drawings
FIG. 1 is a graph showing the particle size distribution of vitamin B2-based acousto-optic nanoparticles VFNS obtained in the examples of the present invention;
FIG. 2 is a schematic diagram showing the stability and dispersibility of vitamin B2-based acousto-optic nanoparticle VFNS according to the present invention;
FIG. 3 is a graph showing the photodynamic behavior of vitamin B2-based acousto-optic (photo) sensitive nanoparticles VFNS obtained in the examples of the present invention in vitro;
FIG. 4 is a graph showing the in vitro sonodynamic performance of vitamin B2-based acousto-optic (photo) sensitive nanoparticles VFNS obtained in accordance with the present invention;
FIG. 5 is an in vitro GSH consumption profile of vitamin B2-based acousto-optic (photo) sensitive nanoparticles VFNS obtained in accordance with the present invention;
FIG. 6 is a graph showing the toxicity evaluation of vitamin B2-based acousto-optic nanoparticle VFNS of the present invention on 4T1 cells under ultrasound conditions;
FIG. 7 is a graph showing the toxicity evaluation of vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS obtained in the example of the present invention on 4T1 cells under the condition of blue light irradiation;
FIG. 8 is a graph showing the ability of vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS of an embodiment of the present invention to generate Reactive Oxygen Species (ROS) in a cell under ultrasound conditions;
FIG. 9 is a graph showing the ability of vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS of the present invention to generate Reactive Oxygen Species (ROS) in cells under blue light irradiation;
FIG. 10 is a graph showing the effect of vitamin B2-based acousto-optic nanoparticle VFNS on 4T1 cell activity under ultrasound conditions;
FIG. 11 is a graph showing the effect of vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS obtained in the example of the present invention on the activity of 4T1 cells under blue light irradiation;
fig. 12 is a graph showing tumor treatment of tumor-bearing mice with vitamin B2-based acousto-optic nanoparticle VFNS obtained in the examples of the present invention.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and completely with reference to the accompanying drawings, in which it is apparent that the embodiments described are only some embodiments of the present invention, but not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
Referring to FIGS. 1-12, a method for preparing vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS composed of polyhydroxy structure of vitamin B2 and Fe 3+ Coordination composition, the reaction equation is shown as follows:
the nanoparticle VFNS specifically comprises the following steps:
step one: the vitamin B210mg is precisely weighed, 100ml of pure water is added for dissolution, and centrifugation is carried out at a speed of 10000rpm for 10 minutes, so that the completely dissolved vitamin B2 mother liquor is obtained.
Step two: 4ml of vitamin B2 mother liquor was precisely measured, 0.37mmol FeCl3.6H2O was slowly added at the same time, and the mixture was stirred at room temperature for 30 minutes by using a magnetic stirrer (rpm=1000 rpm).
Step three: and (3) filling the obtained solution into a dialysis bag, dialyzing for 12 hours by using pure water, collecting the nano-liquid, and freeze-drying the nano-liquid to obtain the vitamin B2-based acousto-optic (photo) sensitive nano-particles VFNS.
The prepared sound (light) sensitive nano-particle VFNS is shown in figure 1, and the stability and dispersibility of the sound (light) sensitive nano-particle VFNS are shown in figure 2.
5 μg/mL VFNS nanoparticle and 1mg/mL 1,3 Diphenylisobenzofuran (DPBF) were dissolved in 3.0mL ultra-pure water, sonicated (ultrasound frequency 1MHz duty cycle-80% power-1.0 w/cm) 2 ) Different times or blue light irradiation (50-100 mW cm) -2 ) For different times, the solutions were tested for uv absorption, respectively. DPBF is a probe for detecting active oxygen, and the absorption of the characteristic peak at 410nm gradually decreases with the generation of active oxygen.
The sonodynamic behavior of the above-described acoustically (photo) sensitive nanoparticle VFNS in vitro is shown in fig. 3 (with 1,3 diphenyl isobenzofuran probe); with the extension of ultrasonic time or illumination time, the absorption value of the characteristic peak of DPBF at 410nm is observed to be gradually reduced, which indicates that the vitamin B2-based acousto-optic (photo) sensitive nano particles VFNS can generate a large amount of active oxygen under the conditions of ultrasonic light and blue light irradiation, and have good in vitro acoustic power and photodynamic performance.
GSH (glutathione) solution (1 mM, 500. Mu.l) and VFNS solution (30. Mu.g/ml, 500. Mu.l) were taken, PBS buffer (2 ml) was added, stirred with a magnetic stirrer (200 rpm, room temperature) and the mixture was centrifuged (10000 rpm,10 min) for 0/2/4/6/18/24 hours, and the supernatant was taken, DTNB (1 mg/ml, 50. Mu.l) was added, and the ultraviolet absorbance spectrum was measured. DTNB [ (5, 5 '-dithiobis (2-nitrobenzoic acid)) ] is commonly used to detect free thiol (-SH), also known as Ellman's reagent, which reacts with GSH to produce TNB and GSSG (412 nm) which stabilize yellow, with the absorbance at 412nm gradually decreasing as GSH is consumed.
The GSH (glutathione) consumption capacity of the above-mentioned acousto-optic (photo) sensitive nanoparticle VFNS in vitro is shown in FIG. 5[ (use of 5,5' -dithiobis (2-nitrobenzoic acid)) ]; with the increase of time, the gradual decrease of absorbance at 412nm can be observed, which indicates that the vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS has a certain GSH (glutathione) consumption capability and has an iron death effect.
Different concentrations of vitamin B2-based acousto (photo) sensitive nanoparticle VFNS were incubated with mouse breast cancer (4T 1) cells for 24h and then tested for cellular activity using the standard MTT method. Vitamin B2-based ultrasound (ultrasound frequency 1MHZ duty cycle-80% power-1.0 w/cm) 2 ) 5min or blue light irradiation (50-100 mW cm) -2 ) 30min, then incubated with 4T1 cells for 24h, to evaluate its therapeutic effect on photodynamic or photodynamic therapy.
The toxicity evaluation of the vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS on 4T1 cells at the above different concentrations is shown in fig. 6 and 7, and it can be observed that the vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS has a certain toxicity on 4T1 cells under the condition of ultrasonic or blue light irradiation.
30 μg/ml of vitamin B2-based acousto (photo) sensitive nanoparticles VFNS were incubated with mouse breast cancer (4T 1) cells for 6h, blue laser (50-100 mW cm -2 ) Irradiation for 10min or ultrasound (ultrasound frequency 1MHz duty cycle-80% power-1.0 w/cm) 2 ) 5 minutes. DCFH-DA probe was added and its reactive oxygen species ROS were detected by laser confocal detection. DCFH-DA (dichlorofluorescein-acetoacetate) is a cell permeable probe for the detection of Reactive Oxygen Species (ROS) in cells.
The ability of the above-described acoustically (photo) sensitive nanoparticle VFNS to generate reactive oxygen species ROS within the cell is shown in fig. 8 and 9 (using DCFH-DA probe); DCFH-DA itself has no fluorescence, can freely pass through cell membrane, is hydrolyzed by esterase to generate DCFH, accumulates in cells, and reactive oxygen species ROS oxidize the DCFH to generate DCF with fluorescence, and green fluorescence intensity is in direct proportion to reactive oxygen species level under 488nm excitation. As shown in fig. 8 and 9, the acousto-optic (photo) sensitive nanoparticle VFNS can generate a large amount of reactive oxygen species ROS under the conditions of ultrasonic and blue light irradiation, which indicates that the nano-particle VFNS has good acousto-optic power and photo-dynamic performance in cells.
30 μg/ml of vitamin B2-based acousto (photo) sensitive nanoparticles VFNS were incubated with mouse breast cancer (4T 1) cells for 24h, blue laser (50-100 mW cm -2 ) Irradiation for 30min or ultrasound (ultrasound frequency 1MHz duty cycle-80% power-1.0 w/cm) 2 ) 5 minutes and further incubation for 24 hours. Cell survival was assessed by Calcein-AM/PI.
The ability of the above-described acousto (photo) sensitive nanoparticle VFNS to affect the size of cell activity is shown in FIGS. 10 and 11 (using Calcein-AM/PI reagent). Calcein-AM (Calcein acetyl methyl ester), a living cell fluorescent dye with cell membrane permeability, is green fluorescent, and only marks living cells; PI can only cross disordered areas of dead cell membranes to reach the nucleus, producing red fluorescence. As shown in fig. 10 and 11, the acousto-optic (photo) sensitive nano particles VFNS can have a significant effect on cell activity under the conditions of ultrasonic and blue light irradiation, which indicates that the nano particles VFNS have good acousto-optic power and photo-power effects on tumor cells.
The graph of the above-mentioned vitamin B2-based acousto (photo) sensitive nanoparticle VFNS on tumor treatment of tumor-bearing mice is shown in fig. 10; 4T1 tumor-bearing mice (tumor size: -50 mm) were selected 3 ) Randomly divided into 4 groups (6 per group):
group (1) is control, PBS alone;
group (2) is a material group (VFNS), tail vein injection of vitamin B2-based acousto-optic (photo) sensitive nanoparticles VFNS alone;
group (3) is a material (VFNS) light therapy group, tail vein injection of vitamin B2-based acousto (photo) sensitive nanoparticles VFNS+blue light (50-100 mW cm) -2 ) 5 minutes/min;
group (4) is a material (VFNS) ultrasound treatment group, tail vein injection is based on vitamin B2 sound (light) sensitive nanoparticle VFNS+ ultrasound condition (ultrasound frequency 1MHZ duty cycle-80% power-1.0 w/cm) 2 ) 5 minutes/min.
The tumors were treated with blue light/ultrasound irradiation 4 hours after tail vein injection, and the above procedure was repeated three times every three days. Tumor volumes were measured and recorded daily. Tumor volume is determined by the following equation: volume = length x width 2 /2。
Compared with the control group, the (2) group only shows a certain tumor inhibiting effect when being injected by tail vein with the vitamin B2-based sound (light) sensitive nano particle VFNS group, which indicates that iron death can play a certain role; group 3 tail intravenous injection of vitamin B2-based acousto (photo) sensitive nanoparticles vfns+blue light irradiation group, treatment effect is weak due to limitation of blue light; the group 4 tail intravenous injection of the vitamin B2-based sound (light) sensitive nanoparticle VFNS+ ultrasonic irradiation group has obviously obtained treatment effect compared with other groups, which shows that the nanoparticle can apply sound (light) power and iron death to tumor treatment in combination, thereby achieving the effect of cooperative treatment.
The present invention is not described in detail in the present application, and is well known to those skilled in the art.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.
Claims (3)
1. A preparation method of vitamin B2-based acousto-optic nanoparticle VFNS is characterized in that: the nano-particle VFNS consists of a polyhydroxy structure of vitamin B2 and Fe 3+ Coordination composition, the reaction equation is shown as follows:
the method specifically comprises the following steps:
step one: precisely weighing FeCl 3 ·6H 2 O；
Step two: precisely weighing vitamin B2, and ultrasonically centrifuging and dissolving with pure water to obtain vitamin B2 water solution;
step three: precisely weighing the vitamin B2 aqueous solution into an ampoule bottle, and slowly adding precisely weighed FeCl under the stirring of a magnetic stirrer 3 ·6H 2 O, continuing stirring for a period of time until the system has obvious Tyndall effect; dialyzing with pure water or concentrating by ultrafiltration, collecting nanometer liquid, and lyophilizing to obtain desired nanometer granule;
in the third step, the concentration of the vitamin B2 aqueous solution is 1-10 mg/100mL, feCl 3 ·6H 2 The amount of the O substance is 0.185mmol to 0.370mmol;
in the second step, the rotation speed of the centrifugal machine is 10000rpm, and the centrifugal time is 10min;
in the third step, the reaction temperature is room temperature, the reaction time is 30-60 min, and the rotating speed of the stirrer is 1000rpm;
in the third step, the dialysis time is 12-24 hours;
in the third step, using an ultrasonic cell grinder to carry out ultrasonic treatment under certain power and time, collecting nano liquid and freeze-drying to obtain the required nano particles; wherein the ultrasonic power is 50-200W, the ultrasonic time is 5-10s, the ultrasonic gap time is 1-5s, and the total duration is 1-10min.
2. Use of vitamin B2-based acousto-optic nanoparticle VFNS obtained by the method of claim 1 in the preparation of antitumor drug therapy.
3. Use of vitamin B2-based acoustical and photosensitive nanoparticle VFNS obtained by the method of preparation according to claim 1 for the preparation of acoustical, photodynamic and iron-death medicaments.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210370030.6A CN114917191B (en) | 2022-04-08 | 2022-04-08 | Preparation method and application of vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210370030.6A CN114917191B (en) | 2022-04-08 | 2022-04-08 | Preparation method and application of vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114917191A CN114917191A (en) | 2022-08-19 |
| CN114917191B true CN114917191B (en) | 2023-07-28 |
Family
ID=82804317
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210370030.6A Active CN114917191B (en) | 2022-04-08 | 2022-04-08 | Preparation method and application of vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114917191B (en) |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101492477B (en) * | 2009-01-21 | 2011-11-09 | 电子科技大学 | Photosensitizer, composite photocatalyst and method for preparing the same |
| CN103585644A (en) * | 2013-11-13 | 2014-02-19 | 苏州大学 | Polyethylene glycol modified magnetic nanoparticle and application thereof |
| CN109602914B (en) * | 2019-01-08 | 2022-05-17 | 扬州大学 | Vitamin B2Modified iron-based nanoenzyme and preparation method and application thereof |
| CN110559302B (en) * | 2019-08-14 | 2023-04-18 | 深圳大学 | Nano diagnosis and treatment agent and preparation method and application thereof |
| CN112791185B (en) * | 2021-01-20 | 2022-07-08 | 广州医科大学 | Nano medicine for treating tumor by combining photo-thermal treatment with iron agent and preparation method thereof |
| CN113209292B (en) * | 2021-05-14 | 2023-04-04 | 中国人民解放军陆军军医大学 | Quercetin nano self-assembly material, preparation method and application |
-
2022
- 2022-04-08 CN CN202210370030.6A patent/CN114917191B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN114917191A (en) | 2022-08-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Xing et al. | Advances and perspectives in organic sonosensitizers for sonodynamic therapy | |
| CN106139144B (en) | A kind of hyaluronic acid decorated gold-Nano carbon balls and the preparation method and application thereof with synergistic antitumor characteristic | |
| Chu et al. | Pluronic-encapsulated natural chlorophyll nanocomposites for in vivo cancer imaging and photothermal/photodynamic therapies | |
| Zhang et al. | One-pot synthesis of hollow PDA@ DOX nanoparticles for ultrasound imaging and chemo-thermal therapy in breast cancer | |
| Zhu et al. | Free radical as a double-edged sword in disease: Deriving strategic opportunities for nanotherapeutics | |
| CN108452303A (en) | It is a kind of to carry double medicine nanometer formulations and preparation method thereof | |
| JP2007277218A (en) | Liposome composition and method for using the same | |
| CN107952072B (en) | Preparation method of drug-loaded and oxygen-loaded hybrid protein nanoparticles, drug-loaded and oxygen-loaded hybrid protein nanoparticles and application | |
| Cao et al. | A multimodal imaging-guided nanoreactor for cooperative combination of tumor starvation and multiple mechanism-enhanced mild temperature phototherapy | |
| CN112076319B (en) | Application of artemisinin and derivatives thereof in preparation of thermo-dynamic therapy sensitizer | |
| CN111840549B (en) | Platinum drug/photosensitizer-loaded protein nanoparticles and preparation method and application thereof | |
| CN112999153B (en) | Nano micelle carrying chemotherapeutic drug/photosensitizer and preparation method and application thereof | |
| CN113559064B (en) | Novel self-oxygen-supply liposome nanoparticle and preparation method and application thereof | |
| Cheng et al. | An effective NIR laser/tumor-microenvironment co-responsive cancer theranostic nanoplatform with multi-modal imaging and therapies | |
| Geng et al. | Oxygen-carrying biomimetic nanoplatform for sonodynamic killing of bacteria and treatment of infection diseases | |
| CN113018267A (en) | Unsaturated fatty acid-photosensitizer co-assembled nanoparticles and construction method and application thereof | |
| WO2021083370A1 (en) | Preparation and use of nanomaterial specifically activating immune system | |
| EP3682899A1 (en) | Compound amphiphilic peptide nanomicelle, preparation and use thereof | |
| CN113117077B (en) | Platinum-based monatomic nanoenzyme for tumor combined treatment and preparation method thereof | |
| CN114917191B (en) | Preparation method and application of vitamin B2-based acousto-optic (photo) sensitive nanoparticle VFNS | |
| CN110251672B (en) | Nano diagnosis and treatment agent and preparation method and application thereof | |
| CN114668841B (en) | Activatable nano photosensitizer and preparation method and application thereof | |
| CN109966513B (en) | Preparation method and application of multifunctional microbubble integrating ultrasonic/fluorescent bimodal imaging and photodynamic therapy/chemotherapy | |
| CN114601925A (en) | Photosensitive nano material jointly modified by hyaluronic acid and RSL3, preparation method and application thereof | |
| CN113230419A (en) | Novel targeted nano-particles based on phycocyanobilin and preparation method thereof |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |