CN115518154A - FeCuNC nano material and preparation and application thereof - Google Patents

FeCuNC nano material and preparation and application thereof Download PDF

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CN115518154A
CN115518154A CN202211178767.4A CN202211178767A CN115518154A CN 115518154 A CN115518154 A CN 115518154A CN 202211178767 A CN202211178767 A CN 202211178767A CN 115518154 A CN115518154 A CN 115518154A
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fecunc
nano material
tumor
annealing
solution
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CN115518154B (en
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祖雄兵
李辉煌
胡姣
邓留
陶娜
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Xiangya Hospital of Central South University
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    • A61K41/00—Medicinal preparations obtained by treating materials with wave energy or particle radiation ; Therapies using these preparations
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • Y02E60/00—Enabling technologies; Technologies with a potential or indirect contribution to GHG emissions mitigation
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    • Y02E60/36—Hydrogen production from non-carbon containing sources, e.g. by water electrolysis

Abstract

The invention belongs to the field of biomedicine. In particular to a FeCuNC nano material and preparation and application thereof. The invention discovers that FeCuNC has strong photothermal effect and rich pseudoenzyme activity. The photothermal effect can further accelerate the decomposition of the over-expressed hydrogen peroxide in the tumor to generate rich active oxygen, and can realize the chemical kinetics and photothermal synergistic treatment, induce the apoptosis of cancer cells and inhibit the growth of the tumor.

Description

FeCuNC nano material and preparation and application thereof
Technical Field
The invention belongs to the technical field of anticancer nano-material development, and relates to a FeCuNC nano-material, a preparation method thereof and application thereof in preparing a photothermal therapy and chemodynamics therapy combined anticancer drug based on photothermal effect and mimic enzyme activity.
Background
Cancer remains one of the most serious diseases in humans, and thus the development of highly effective anticancer medical strategies is urgently needed. To date, conventional single modality therapies still suffer from poor bioavailability, impaired target specificity, systemic and organ toxicity, and the like. Combination therapy is one of the most promising approaches to address these problems due to its high efficacy and low risk of recurrence. Early combination therapies included lipid and polymer nanocapsules, two or three drugs, but their performance was largely dependent on the drug loading capacity of the different drugs. So far, there is an increasing interest in new medical applications of two-dimensional nanomaterials with extraordinary physicochemical properties, and research for developing two-dimensional nanocarriers for multimodal nanomedicine has proliferated. E.g. graphene, moO x ，WS 2 And MoS 2 Have been used to construct combination therapy platforms that exhibit outstanding performance in cancer therapy. However, developing new strategies of multifunctional two-dimensional nanomaterials with higher loading capacity for synergistic combination therapies remains a major challenge.
Hydrogen peroxide (H) 2 O 2 ) Plays an important role in many physiological and pathological processes, especially H associated with various cancers 2 O 2 Taking advantage of the imbalance, reactive oxygen species (ROS, including singlet oxygen, superoxide anions, and hydroxyl radicals) can be generated, causing oxidative damage to cancer cells. Nanocatalysts involved in chemokinetic therapy (CDT) can promote H in the tumor microenvironment through biomimetic catalytic processes 2 O 2 Converted into highly toxic ROS. Nanocarbon suspensions are important nanomarkers for clinical diagnosis of lymph node biopsies, and carbon materials show great potential in Infrared (IR) thermography and photothermal therapy (PTT) due to their availability of photothermal properties. Therefore, the specific diatomic sites on the optimized structure of the nano-carbon particles are expected to be effective for activating H 2 O 2 Thereby facilitating high-precision diagnosis and ablation of metastatic Sentinel Lymph Nodes (SLNs).
Disclosure of Invention
In order to realize the efficient identification and treatment of the tumor at the lymph node, the hydrogen peroxide in the tumor microenvironment is fully utilized, and the problems of poor biological safety, poor drug resistance, single functionality and the like of the existing anti-tumor nano material are solved. The invention mainly aims to provide a FeCuNC nano material which is a hollow nitrogen-doped carbon nanosphere and has Fe and Cu active sites. The material can be used for preparing anti-tumor drugs; aims to realize the combination of photothermal therapy and chemokinetic therapy with anti-cancer drugs, realize the recognition of sentinel lymph node metastasis and effectively inhibit tumor metastasis and recurrence.
The hydration diameter of the FeCuNC nano material is 100-300 nm.
The second purpose of the invention is to provide a preparation method of FeCuNC nanometer material, which comprises the following steps:
adding tetraethyl orthosilicate into a mixed solution of ammonia water and absolute ethyl alcohol or propyl alcohol to obtain template SiO 2 (ii) a Adding a Dopamine (DA) aqueous solution into the mixed solution, continuously stirring, then adding iron (II) acetylacetonate or ruthenium acetylacetonate, continuously stirring, centrifuging, washing and drying to obtain Fe-PDA-SiO 2 ；Fe-PDA-SiO 2 Annealing in protective atmosphere to obtain FeNC; feNC, urea and copper (II) phthalocyanine (CuPc) are uniformly mixed and then annealed in a protective atmosphere to obtain the FeCuNC nano material.
Further, the preparation method of the FeCuNC nano material,
the concentration of ammonia water in the initial solution is 15-35 wt%.
The volume ratio of the ammonia water to the absolute ethyl alcohol or the propyl alcohol is 1:20 to 1:40.
tetraethyl orthosilicate, and the volume ratio of the tetraethyl orthosilicate to the mixed solution of ammonia water and absolute ethyl alcohol or propanol is 1:15 to 1:30.
the concentration of the dopamine solution is 2 to 5 weight percent; the volume ratio of the dopamine solution to the mixed solution is 1:5 to 1:8.
the mass ratio of iron (II) acetylacetonate or ruthenium acetylacetonate to dopamine is 1: 1.5-1: 3.
Fe-PDA-SiO 2 annealing the powder in nitrogen at a heating rate of 3-8 ℃ for min -1 (ii) a The annealing temperature is 700-800 ℃, and the annealing time is 1.5-2.5 h.
FeNC, urea and copper (II) phthalocyanine in a mass ratio of 1:40:0.25 to 1:60:0.3.
FeNC, urea and copper (II) phthalocyanine are mixed evenly and then annealed in nitrogen, and the heating rate is 2.5-3.5 ℃ for min -1 (ii) a The annealing temperature is 550-650 ℃, and the annealing time is preferably 1.5-2.5 h.
The third purpose of the invention is to provide the application of the FeCuNC nano material in decomposing hydrogen peroxide to generate active oxygen or in preparing a preparation for decomposing hydrogen peroxide to generate active oxygen.
Furthermore, the FeCuNC nano material can accelerate the decomposition of hydrogen peroxide and enhance the generation of active oxygen by irradiating near-infrared II-region light.
Furthermore, the power of the illumination is 0.5-2.2W cm -2 The illumination time is 0.5-10 min, and the concentration of illumination treatment is 0.5-50 ppm.
Further preferably, the power of the light irradiation is 1.2-2.0W cm -2 The illumination time is 2-5 min, and the illumination treatment concentration is 10-40 ppm.
Further, mixing FeCuNC and the target modifier, stirring, and centrifugally washing to obtain a FeCuNC @ CM nanosphere target material; preferably, the targeted modifier is extracted cell membrane, including cancer cell membrane, leucocyte membrane; the concentration ratio of FeCuNC and cell membrane in the mixed system solution is 0.5-2.
The targeting receptor is protein, and the targeting of different cancer cells is realized according to active molecules on the surfaces of different cell membranes.
The fourth purpose of the invention is to provide the application of the FeCuNC nanometer material in preparing anti-tumor drugs; in particular to the application in the anti-tumor medicine with photo-thermal enhancement.
The fifth purpose of the invention is to provide an anti-tumor drug, which comprises the FeCuNC nano material.
The FeCuNC nano material has catalase mimic enzyme activity.
Further, the FeCuNC nano material also has peroxidase mimic enzyme activity, and can catalyze H 2 O 2 Generating active oxygen to realize chemical dynamic therapy.
Furthermore, the FeCuNC nano material can generate heat through irradiating the FeCuNC nano material by near infrared II area light; preferably, the power of the illumination is 0.5-2.2W cm -2 The time of illumination is 0.5-10 min, and the concentration of illumination treatment is 0.5-50 ppm.
Furthermore, the FeCuNC nano material is irradiated by near-infrared II-region light, so that the decomposition of hydrogen peroxide can be accelerated, the generation of active oxygen is enhanced, and the chemical kinetics treatment is further cooperated; preferably, the power of the illumination is 1.2-2.0W cm -2 The time of light irradiation is 2-5 min, the concentration of light irradiation is 10-40 ppm, and the concentration of hydrogen peroxide is 0.05-50 mM.
Furthermore, the FeCuNC nano material can generate heat by irradiating the FeCuNC nano material by near-infrared II-region light, and has certain light circulation heat generation capacity; the power of the illumination circulation is 1-2.2W cm -2 The time of light irradiation is 40-200 min, and the concentration of light irradiation treatment is 10-50 ppm.
The FeCuNC nano material is used for preparing the antitumor drugs, has excellent antitumor activity, and can effectively kill tumor cells and ablate tumors; can realize the recognition of sentinel lymph node metastasis and inhibit tumor metastasis and recurrence.
The research of the invention finds that the nano-sized FeCuNC material can effectively generate active oxygen under the condition of illumination or no illumination.
The research of the invention finds that the material prepared by the preparation method has better biocompatibility, better photothermal promotion active oxygen release and stronger mimic enzyme activity, and better tumor effect.
The tumors of the present invention may be benign and/or malignant tumors.
The invention is used for applying the medicine to the mice by a tail vein injection method.
For example, in the using process, feCuNC @ CM nanospheres can be dispersed by adopting physiological saline; the resulting dispersion solution is then administered by intravenous injection. The preferable dosage is 1.5-3 mg kg 1 . The dose of hollow fecucc @ cm nanospheres preferably applied per subject (e.g. mouse) is 50ug.
Preferably, the FeCuNC @ CM nano material is not less than the pharmaceutically effective amount; preferably, the composition also comprises pharmaceutically acceptable auxiliary materials; preferably, in any pharmaceutically acceptable dosage form; preferably, it is a pharmaceutically acceptable injection formulation; further preferred is a pharmaceutically acceptable topical injection formulation.
The research of the invention finds that the FeCuNC @ CM nano material can promote the formation of active oxygen in tumor cells based on a photo-thermal mode; in addition, feCuNC @ CM nanomaterials have good peroxidase-mimetic enzyme activity, e.g., they have good peroxidase activity, catalyzing H 2 O 2 Active oxygen is generated.
The research of the invention discovers that the FeCuNC @ CM nano material can catalyze hydrogen peroxide to generate active oxygen in situ at the tumor tissue through photo-thermal promotion and cooperation with chemokinetic treatment, so that the active oxygen content at the biological tissue is improved. Compared with the traditional medicine molecules, feCuNC @ CM is recycled to generate active oxygen, so that the medicine property is improved. Secondly, the FeCuNC @ CM nano material has good biocompatibility, so that the FeCuNC @ CM nano material shows good biological safety for treating the breast cancer of the mouse.
Has the beneficial effects that:
(1) The developed FeCuNC @ CM nano material has good anti-tumor activity;
(2) The developed FeCuNC @ CM nano material achieves the effects of killing tumor cells and improving the anti-tumor effect through a photo-thermal combination chemical kinetics reaction mechanism;
for example, fecucc @ cm nanomaterials can catalyze hydrogen peroxide to generate active oxygen in situ at tumor tissue by photothermal promotion in an organism in cooperation with chemokinetic treatment, thereby increasing the active oxygen content at the biological tissue. Enhancing active oxygen content, and killing tumor cells by oxidation.
In addition, the good biocompatibility of the FeCuNC @ CM nano material enables the FeCuNC @ CM nano material to show good biological safety for treating the breast cancer of the mouse.
(3) The developed FeCuNC @ CM nano material realizes the recognition of sentinel lymph node metastasis through the carbon sphere imaging function of the carbon suspension, can realize the original treatment of lymph node metastasis through cell membrane targeting and combination of photo-thermal, and further effectively inhibits the metastasis and recurrence of tumors.
(4) The preparation method of the FeCuNC @ CM nano material is simple, mild in reaction condition, low in cost, large in application prospect and capable of being prepared in a large scale.
Drawings
FIG. 1 is a transmission electron micrograph (TEM image), an X-ray diffraction chart (XRD image), a high-resolution transmission electron micrograph (HRTEM image) and an energy spectrum (EDS) distribution diagram of a hollow nitrogen-doped carbon nanosphere FeCuNC nanomaterial prepared in example 1;
wherein (a) is a TEM image; (b) is an XRD pattern; (c) HRTEM image; and (d) is an EDS distribution diagram.
FIG. 2 shows the photothermal effect and the detection result of a mimic enzyme of a hollow FeCuNC nano material;
(a) A graph of simulated peroxidase activity for detection; (b) is a photo-thermal effect graph for detecting different FeCuNC concentrations; and (c) is a graph for detecting FeCuNC photothermal effect under different powers.
FIG. 3 is a study of the antitumor effect at the cell level;
(a) Mapping the endocytosis effect of cells on FeCuNC at different times; (b) Different cell types were incubated for 24h in fecucc treatment for cell viability maps; (c) Fluorescence images of intracellular reactive oxygen species stained by DCFH-DA after different treatments; (d) Flow-through treatment of intracellular reactive oxygen species stained with DCFH-DA following different treatments; (e) After different treatments, mouse breast cancer cells were incubated for 24h in 4T1 for cell viability;
control, NC, NC + H in FIG. 3 2 O 2 ，FeNC，FeNC+H 2 O 2 ，FeCuNC，FeCuNC+H 2 O 2 ,FeCuNC+Laser，FeCuNC+H 2 O 2 + Laser is the product of cell membrane targeted modifierA compound (I) is provided.
FIG. 4 is a study of the effect of tumor treatment in vivo at animal level;
(a) The body weight of the mouse at different time under different conditions is shown; (b) Is a graph of tumor volume of mice at different times under different conditions; (c) is a visual map of the tumor of the mice at day 18 under different conditions; (d) The temperature rise effect of the mouse in vivo photo-thermal at different time under different conditions is shown; (e) And (3) a temperature rise curve graph of the mouse in vivo photo-thermal at different times under different conditions.
Detailed description of the invention
The present invention is further illustrated by the following specific examples. These examples are to be construed as merely illustrative and not limitative of the remainder of the disclosure in any way whatsoever. After reading the disclosure of the present invention, various changes and modifications of the present invention based on the principle of the present invention also fall into the scope of the present invention as defined in the appended claims.
Example 1
Preparation of hollow FeCuNC @ CM nanospheres:
ethanol (70 mL), H 2 O(10mL)、NH 3 ·H 2 O (3 mL, 30wt%) was mixed and stirred for 10 minutes. Then, tetraethyl orthosilicate (3.5 mL) was added and stirred for an additional 15min. 10mL of an aqueous solution containing 0.5g of dopamine was added and stirred for 6h. Then, 254mg of iron (II) acetylacetonate was added thereto, and the mixture was stirred for 12 hours. The product Fe-PDA-SiO 2 Centrifuging, washing with ethanol for 3 times, and drying at 70 deg.C. Fe-PDA-SiO 2 Under nitrogen at 5 deg.C for min –1 Annealing at 750 ℃ for 2h. The resulting black powder was then washed with HF (10%) for 24h to give FeNC. Then, 20mg of FeNC, 1g of urea and 5mg of copper (II) phthalocyanine (CuPc) were pulverized and mixed uniformly. Then in N 2 In the atmosphere, at 3 ℃ for min –1 Annealing at 600 ℃ for 2h to obtain FeCuNC, and adding FeCuNC (1mL, 1mg mL) –1 ) And extracted 4T1-Luc cell membrane (10. Mu.L, 10mg mL) –1 ) Mixing, stirring for 12h, and centrifuging to obtain FeCuNC @ CM.
From the parts (a), (c) and (d) of fig. 1, it can be known that the fecucc nanosphere is a uniform hollow nitrogen-doped carbon nanosphere.
Example 2
Testing of the performance of hollow-core fecucc nanospheres (prepared in example 1) to mimic peroxidase activity: the experiments were divided into the following groups:
(1) Non-modified nitrogen-doped carbon Nanospheres (NC); (2) iron-modified nitrogen-doped carbon nanospheres (FeNC); (3) The method comprises the following steps of (1) modifying nitrogen-doped carbon nanospheres (FeCuNC) by iron and copper at double sites; using 3,3, '5,5' -tetramethyl benzidine (TMB) as probe, in HAc-NaAc buffer solution and H 2 O 2 In the presence of an aqueous solution, H is measured 2 O 2 And (4) activating efficiency. To HAc-NaAc buffer (100. Mu.L, pH =3.5,0.1M) was added FeCuNC, feNC or NC (5. Mu.L, 1mg mL) in sequence –1 )、H 2 O 2 Aqueous (100. Mu.L, 100 mM) and TMB (100. Mu.L, 1 mM) alcoholic solution. The absorbance of the mixed solution at 652nm was recorded with an ultraviolet spectrophotometer.
From the part (a) of fig. 2, it can be seen that the ultraviolet absorption intensity of the fecucc nanosphere is the highest, demonstrating that the fecucc nanosphere has the strongest ability to generate active oxygen.
Example 3
Testing of photo-thermal properties of hollow fecucn nanospheres (prepared in example 1):
the experiments were divided into two subgroups according to different variable parameters:
(1) Temperature rise change of FeCuNC with same power and different concentrations
FeCuNC nanospheres with different concentrations are irradiated for 10min by 808nm laser, and the temperature of the FeCuNC nanospheres is monitored and recorded by a thermal infrared imager (FLIR C2) every 30 s. Wherein the different concentrations include: 0,10, 17.5, 25, 37.5, 50ppm, power fixed at 1.0W cm –2 。
(2) FeCuNC temperature rise change with same concentration and different power
FeCuNC nanospheres (concentration fixed at 25 ppm) of the same concentration were irradiated with 808nm laser for 10min, and their temperature was monitored and recorded every 30s by a thermal infrared imager (FLIR C2). Wherein the different powers include: 0.5,1.0,1.4,1.8,2.2W cm –2 。
From fig. 2 (b), it is known that the light irradiation effect of the fecucc nanospheres increases with the increase of the concentration and the temperature changes, and the maximum concentration of 50ppm can increase to 25 ℃ within 10 min. From fig. 2 (c), it can be known that the illumination effect of the fecucnc nanosphere increases with the increase of the power, the temperature change is continuously increased, and the maximum temperature rise change can reach 60 ℃.
Example 4
Detection of uptake capacity of hollow fecucc @ cm nanomaterial (prepared in example 1) at cell level: in the invention, rhodamine B (RhB, 1mg mL) -1 1 mL) and FeCuNC (1 mg mL) -1 1 mL) was mixed and stirred overnight, centrifuged at 8000rpm for 10min, the resulting solution was washed to supernatant to obtain RhB-modified FeCuNC (RhB-FeCuNC) and then 4T1 cells (10 per well) were added 4 Individual cell) with RhB-FeCuNC (0.025 mg mL) -1 100 uL) were incubated in a 96-well plate for different times of 0, 0.5,1.0 and 2.0h to observe the effect of cellular uptake.
From fig. 3a, it can be seen that the fluorescence intensity of intracellular rhodamine B (RhB) is gradually increased with the time, indicating that fecunc @ cm is effectively endocytosed by 4T1 cells, and the more nanospheres are endocytosed with the time.
Example 5
Detection of toxicity at cell level of hollow fecucc @ cm nanomaterial (prepared in example 1):
the experiments were divided into two subgroups according to different variable parameters:
(1) Incubating different cells in different cell environments for the same time, and detecting cell activity
The invention relates to a method for preparing FeCuNC @ CM nano material (0, 25, 50, 75, 100 ppm) with different concentrations, cancer cells (mouse breast cancer cells 4T1; human non-small cell lung cancer cells A549) and normal cells (mouse embryo fibroblast 3T3; human umbilical vein endothelial cells HUEVC, mouse epithelial fibroblast L929) (each hole is 10) 4 Individual cells, cell purchase Yu Xiangya medical school) were incubated for 24h.
As shown in FIG. 3 (b), after incubation of FeCuNC @ CM nanomaterial with different concentrations for 24h with different normal cells, the cell activity has no obvious change, and after incubation of FeCuNC @ CM nanomaterial with cancer cells for 24h, the cell survival rate has a certain decrease, which indicates that FeCuNC @ CM nanomaterial has no obvious cytotoxicity to normal cells, has good biocompatibility, and has a certain killing ability to cancer cells.
(2) Incubation of mouse breast cancer cell 4T1 under different treatment groups of FeCuNC for the same time, detection of cell activity the invention combines different treatment groups (concentration is 25 ppm) with the same concentration and mouse breast cancer cell 4T1 (each hole is 10 ppm) 4 Individual cells, cell purchase Yu Xiangya medical school) were incubated for 24h. The different treatment groups were: control, NC @ CM + H 2 O 2 ，FeNC@CM，FeNC@CM+H 2 O 2 ，FeCuNC@CM，FeCuNC@CM+H 2 O 2 ,FeCuNC@CM+Laser，FeCuNC@CM+H 2 O 2 +Laser。
H in each of the above groups 2 O 2 The concentration is 50ppm; the illumination conditions of the Laser are all 1.0W cm –2 ，5min。
From fig. 3 (e), it can be seen that after incubation of the nanomaterials and the mouse breast cancer cells in different treatment groups for 24h, the cell activity is significantly changed, and after incubation of the nanomaterials and the mouse breast cancer cells for 24h, the cell survival rate is reduced to a certain extent, and the cytotoxicity of the group combining fecucc @ cm with hydrogen peroxide and photothermal therapy is the strongest, which indicates that the cancer cells are extremely strongly killed by the chemokinetic combined photothermal therapy.
Example 6
Detection of the ability of hollow fecucc @ cm nanomaterials (prepared in example 1) to generate reactive oxygen species at the cell level:
the production of reactive oxygen species in vivo is illustrated in two different ways:
(1) Qualitatively, the description explains the generation of active oxygen in vivo
ROS are produced in cells and can be detected by 2',7' -chlorofluorescein diacetate (DCFH-DA). The present invention relates to 4T1 cells (2X 10) 4 per well) were placed in 24-well plates and incubated for 4h under different conditions. Wherein H 2 O 2 Is 20mM; the final concentration of NC @ CM, feNC @ CM and FeCuNC @ CM was 25ppm; the illumination power is 0.5W cm -2 (ii) a The illumination time is 2min; DCFH-DA was mixed with the medium to a final concentration of 20mM.
As can be seen in FIG. 3 (c), the DCFH-DA probe was oxidized by ROS to produce 2,7-Dichlorofluorescein (DCF), which is green-fluorescent. All groups produced green fluorescence under fecucc @ cm conditions. The group with the strongest fluorescence intensity was FeCuNC @ CM combined with hydrogen peroxide and photothermal, indicating that the chemokinetic and photothermal treatments produced a synergistic effect of ROS.
(2) Quantitatively illustrating the generation of active oxygen in vivo
The present invention uses 4T1 cells (2X 10) 4 per well) were placed in 24-well plates and after incubation for 4h under different conditions, all cells were collected by trypsinization and detected by flow cytometry. Wherein H 2 O 2 Is 20mM; the final concentration of NC @ CM, feNC @ CM and FeCuNC @ CM was 25ppm; the illumination power is 0.5W cm -2 (ii) a The illumination time is 2min; DCFH-DA was mixed with the medium to a final concentration of 20mM.
From the flow statistics of FIG. 3 (d), it can be seen that all groups produced green fluorescence under the conditions of FeCuNC @ CM and under the action of ultrasound. The group with the strongest green fluorescence shift intensity was FeCuNC @ CM, which was combined with hydrogen peroxide and photothermal, and the synergistic effect was also confirmed.
Example 7
Detection of photothermal conversion capability of animal layer of hollow fecucc @ cm nanomaterial (prepared in example 1): 4T1 cells are taken as model cells, BALB/C female mice are taken as model mice, a mouse subcutaneous tumor model is constructed, and FeCuNC @ CM provided by the invention is taken as an anti-tumor material. The experiment was performed by injecting the material into mice (2.5 mg kg) via tail vein injection, wherein 2 groups of experiment groups were set, i.e., a) control and b) FeCuNC @ CM -1 ). Recording the temperature change of the tumor of the mouse every 30s after 24h, wherein the illumination power is 1.0W cm -2 The light recording time was 5min.
From FIG. 4 (d), it can be seen that the temperature of the tumor of the FeCuNC @ CM group mice is significantly changed, which illustrates the targeting property and high photothermal conversion efficiency of the FeCuNC @ CM nano material. From FIG. 4 (e), it is known that the temperature of the tumor in FeCuNC @ CM group mice can be raised from 34 ℃ to 56 ℃ with a temperature change of about 22 ℃.
Example 8
Detection of anti-tumor capacity of hollow FeCuNC @ CM nanomaterial (prepared in example 1) at animal level: constructing a mouse by using 4T1 cells as model cells and a BALB/C female mouse as a model mouseThe mouse subcutaneous tumor model adopts FeCuNC @ CM provided by the invention as an anti-tumor material. This experiment set up 5 experimental groups (5 mice per group): a) control, b) NC @ CM, c) FeNC @ CM, d) FeCuNC @ CM, e) FeCuNC @ CM + NIR, the material was injected into mice by tail vein injection (2.5 mg kg/kg) -1 ). Wherein the illumination power is 1.0W cm -2 The illumination time is 5min, and the observation is continuously carried out for 18 days. (the body weight and tumor volume of the mice were recorded every two days during this period)
From FIG. 4 (a), it can be seen that there was no significant change in body weight of the mice, indicating that FeCuNC @ CM had no significant toxicity; as can be seen from fig. 4 (b) and (c), the tumors of the mice in the control, nc @ cm, fenic @ cm, fecucnc @ cm group were not significantly inhibited, whereas the tumors of the mice in the fecucnc @ cm + near infrared light (NIR) group were significantly reduced after 18 days, and the tumor inhibition rate was high, indicating that the fecucc @ cm + near infrared light (NIR) group had an excellent anti-tumor effect.

Claims (10)

1. A FeCuNC nano material is characterized in that the material is a hollow nitrogen-doped carbon nanosphere and has Fe and Cu active sites.
2. FeCuNC nanomaterial according to claim 1, characterized in that the hydrated diameter of the FeCuNC nanomaterial is between 100 and 300nm.
3. A preparation method of FeCuNC nano material is characterized by comprising the following steps:
adding tetraethyl orthosilicate into a mixed solution of ammonia water and absolute ethyl alcohol or propyl alcohol to obtain template SiO 2 (ii) a Adding a dopamine aqueous solution into the mixed solution, continuously stirring, then adding ferric acetylacetonate or ruthenium acetylacetonate, continuously stirring, centrifuging, washing and drying to obtain Fe-PDA-SiO 2 ；Fe-PDA-SiO 2 Annealing in protective atmosphere to obtain FeNC; and mixing FeNC, urea and copper phthalocyanine uniformly, and annealing in a protective atmosphere to obtain the FeCuNC nano material.
4. The method for preparing FeCuNC nano-material according to claim 3, characterized in that,
the concentration of ammonia water in the initial solution is 15-35 wt%;
the volume ratio of the ammonia water to the absolute ethyl alcohol or the propyl alcohol is 1:20 to 1:40;
tetraethyl orthosilicate, and the volume ratio of the tetraethyl orthosilicate to the mixed solution of ammonia water and absolute ethyl alcohol or propanol is 1:15 to 1:30, of a nitrogen-containing gas;
the concentration of the dopamine solution is 2 to 5 weight percent; the volume ratio of the dopamine solution to the mixed solution is 1:5 to 1:8;
the mass ratio of ferric acetylacetonate or ruthenium acetylacetonate to dopamine is 1: 1.5-1: 3;
Fe-PDA-SiO 2 annealing the powder in nitrogen at a heating rate of 3-8 ℃ for min -1 (ii) a The annealing temperature is 700-800 ℃, and the annealing time is 1.5-2.5 h;
FeNC, urea and copper phthalocyanine in a mass ratio of 1:40:0.25 to 1:60:0.3;
FeNC, urea and copper phthalocyanine are mixed evenly and then annealed in nitrogen, and the heating rate is 2.5-3.5 ℃ for min -1 (ii) a The annealing temperature is 550-650 ℃, and the annealing time is preferably 1.5-2.5 h.
5. Use of FeCuNC nanomaterials according to any one of claims 1 to 4 for decomposing hydrogen peroxide to generate active oxygen or for preparing a formulation for decomposing hydrogen peroxide to generate active oxygen.
6. The use of FeCuNC nanomaterials of claim 5, wherein irradiation of the FeCuNC nanomaterials by near infrared region II light accelerates decomposition of hydrogen peroxide, enhancing production of active oxygen.
7. The use of FeCuNC nanomaterial according to claim 6, characterized in that the power of illumination is 0.5-2.2W cm -2 The illumination time is 0.5-10 min, and the concentration of illumination treatment is 0.5-50 ppm.
8. The application of the FeCuNC nano material as claimed in claim 6, wherein the FeCuNC is mixed with the target modifier, stirred and centrifugally washed to obtain the FeCuNC @ CM nanosphere target material; preferably, the targeted modifier is extracted cell membrane, including cancer cell membrane, leucocyte membrane; the concentration ratio of FeCuNC and cell membrane in the mixed system solution is 0.5-2.
9. The use of FeCuNC nanomaterials according to any one of claims 1 to 4 for the preparation of anti-tumor drugs; in particular to the application in the anti-tumor medicine with photo-thermal enhancement.
10. An antitumor drug comprising the FeCuNC nanomaterial according to any one of claims 1 to 4.
CN202211178767.4A 2022-09-27 2022-09-27 FeCuNC nano material, preparation and application thereof Active CN115518154B (en)

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