Preparation method and application of non-natural chiral phenylalanine dipeptide modified arsenene nano material

Abstract

The invention discloses a preparation method and application of a non-natural chiral phenylalanine dipeptide modified arsenene nano material. The preparation method comprises the following steps: (1) dispersing phenylalanine dipeptide and an arsenene nano material in a solvent to obtain a dispersion liquid; and (2) in an ice-water bath, carrying out ultrasonic treatment on the dispersion liquid to carry out a reaction, after the reaction is completed, carrying out rotary evaporation to remove the solvent, washing with water, and centrifugally collecting the precipitate to obtain the phenylalanine dipeptide modified arsenene nano material. The non-natural chiral phenylalanine dipeptide modified arsenene nano-material prepared by the invention has good transmembrane transport promotion and tumor killing ability, can be used for preparing malignant tumor drugs, and is expected to provide a scientific basis for rational design and function optimization of an intelligent chiral delivery system.

Description

Technical Field

[0001] The present invention relates to the technical field of nanomaterials, and particularly to a preparation method and application of an arsenene nanomaterial modified with unnatural chiral phenylalanine dipeptide. Background Art

[0002] In eukaryotes, D-nucleotides, L-amino acids, and L-phospholipids are the basic units that make up organisms. Their unique stereochemical characteristics make chirality control an indispensable key factor in nanomedicine design and biomolecule modification. In December 2024, 38 international scholars jointly issued a warning in the journal *Science*, calling for a halt to the creation of mirror life because these mirror organisms may bypass the immune defense mechanisms of natural organisms, thus posing a fatal infection risk to humans, animals, and plants. The controversy caused by mirror life makes it particularly urgent to explore the impact of the chirality differences of bioactive molecules on the mechanism in nanomaterial modification strategies.

[0003] In recent years, mirror bioactive molecules such as D-amino acids and D-peptides have been widely used for the modification of nanomaterials. Chiral nanomaterials have been proven to have strong optical activity and self-assembly ability, and have shown excellent performance in fields such as optoelectronics, sensors, and enantioselective catalysis. The surface anchoring of chiral ligands and chiral molecules has also been proven to affect life processes such as the cytotoxicity, cellular uptake, and cell adhesion of inorganic nanomaterials, and thus act on their biological effects at the gene and metabolic levels. However, there is still a lack of a clear understanding of how the stereochemical differences in these surface modifications affect the mechanism of action of inorganic nanomaterials.

[0004] Phenylalanine dipeptide is a naturally occurring dipeptide and a key assembly unit for constructing biofunctional nanomaterials. In recent years, it has been widely used in the biomedical field, such as sensing detection, drug delivery, and tissue repair, and has shown excellent theranostic performance. In April 2022, the research group of Bin Xu designed and synthesized a peptide thioester molecule composed of amino acid ethyl thioester and D-type phenylalanine dipeptide, providing a new strategy for the development of novel anti-tumor drugs. The rich π electron cloud of phenylalanine dipeptide provides a structural basis for the coupling modification of nanomaterials, and provides feasibility for the surface modification of its nanomaterials.

[0005] Arsenic trioxide (ATO), as a typical representative of arsenic-containing drugs, has become a first-line therapeutic drug in clinical practice due to its excellent efficacy in the treatment of acute promyelocytic leukemia (APL). Research shows that the complete remission rate of ATO in treating APL patients is as high as 83%-95%, and this remarkable achievement has attracted extensive attention in the scientific community. However, while arsenic-containing drugs show high efficiency in treatment, they also have the defect of high systemic toxicity, and this contradiction seriously restricts their clinical application effect in the treatment of solid tumors.

[0006] To solve the problem of high-dose-dependent toxicity of arsenic, Chinese invention patent CN201510269043.4 discloses a method for loading arsenic agents, its preparation method and application, using surface-modified silica nanomaterials as carriers to deliver arsenic-containing drugs. Academician Tan Weihong, Professor Martin and other teams have developed many excellent multifunctional arsenic drug delivery systems by surface modification with bioactive molecules. For example, the article Smart human serum albumin-As2O3 nanodrug with self-amplified folate receptor-targeting ability for chronic myeloid leukemia treatment published in Angew Chem Int Ed Engl in 2017, a multifunctional drug delivery system of arsenic trioxide developed with glutathione-pretreated folic acid (FA)-labeled human serum albumin (HSA) and a ferritin-delivered arsenic nanodrug system. In addition to the transformation based on arsenic trioxide, the novel single-element two-dimensional nanomaterial arsenene has also been proven to have excellent optoelectronic properties and loading capacity, and can be functionalized by non-covalent interactions. Its original structure and the modified nanoplatform have both been proven to be able to effectively load chemotherapeutic drugs, photodynamic therapy drugs and immunotherapy drugs, and show excellent anti-tumor effects. However, there is still a lack of systematic research reports on the influence of biomolecular chirality regulation on the biological functions and action mechanisms of arsenic nanomaterials.

[0007] Therefore, it is of great significance to develop a novel arsenic-containing drug delivery system based on chiral amino acids / peptides to improve the treatment effect on tumors. Summary of the Invention

[0008] Aiming at the deficiencies of the prior art, the present invention provides a preparation method and application of an arsenene nanomaterial modified with unnatural chiral phenylalanine dipeptide. The arsenene nanomaterial modified with unnatural chiral phenylalanine dipeptide prepared by the present invention has good ability to promote transmembrane transport and tumor killing, and can be used to prepare drugs for malignant tumors, and is expected to provide a scientific basis for the rational design and functional optimization of intelligent chiral delivery systems.

[0009] The technical solution of the present invention is as follows:

[0010] A preparation method of an arsenicene nanomaterial modified with a non-natural chiral phenylalanine dipeptide, the preparation method comprising the following steps:

[0011] (1) Dispersing phenylalanine dipeptide and arsenicene nanomaterial in a solvent to obtain a dispersion;

[0012] (2) Under an ice-water bath, subjecting the dispersion to ultrasonic treatment for reaction. After the reaction is completed, removing the solvent by rotary evaporation, then washing with water and centrifuging to collect the precipitate, thereby obtaining the phenylalanine dipeptide-modified arsenicene nanomaterial.

[0013] Further, in step (1), the phenylalanine dipeptide is a D-type phenylalanine dipeptide; the mass ratio of the phenylalanine dipeptide to the arsenicene nanosheet is 5-6:3.

[0014] Further, in step (1), the solvent is any one of methanol and ethanol; the concentration of the phenylalanine dipeptide in the solvent is 0.2-1 mg / mL.

[0015] Further, in step (2), the power of the ultrasonic treatment is 450-630 W, and the time is 25-30 min.

[0016] Further, in step (2), the temperature of the rotary evaporation is 36-40 °C, and the time is 0.5-1 h.

[0017] Further, in step (2), the rotation speed of the centrifugation is 11000-14000 rpm, and the time is 5-10 min.

[0018] Further, in step (2), the thickness of the phenylalanine dipeptide-modified arsenicene nanomaterial is 20-30 nm.

[0019] An application of the arsenicene nanomaterial modified with a non-natural chiral phenylalanine dipeptide prepared by the above preparation method, wherein the arsenicene nanomaterial is used for preparing a drug for treating malignant tumors.

[0020] The beneficial technical effects of the present invention are as follows:

[0021] The present invention is the first to use phenylalanine dipeptide to modify arsenene nanomaterials, and synthesizes an arsenene nanomaterial modified with a mirror-image biological molecule, D-phenylalanine dipeptide, which has not been reported. First, by optimizing the types of amino acids or dipeptides that modify the arsenene nanomaterials, the present invention discovered that phenylalanine dipeptide is a membrane-destructive dipeptide that helps improve the transmembrane transport of arsenic nanomaterials; at the same time, based on the optimization of the dosage of phenylalanine dipeptide and arsenene nanomaterials, the prepared phenylalanine dipeptide-modified arsenene nanomaterials maintains a large aspect ratio, does not form a tubular structure, and retains a high surface energy. Secondly, D-phenylalanine dipeptide is a non-natural chiral bioactive molecule that is not easily degraded by intracellular proteases, which contributes to the accumulation of arsenic and enhances the killing and treatment of tumors. It is expected to provide a scientific basis for the rational design and functional optimization of intelligent chiral delivery systems. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 Transmission electron micrographs of AS@DD-FF prepared in Example 1 of the present invention and AS@LL-FF prepared in Comparative Example 1.

[0023] Figure 2 Atomic force electron microscopy images of AS@DD-FF prepared in Example 1 of the present invention and AS@LL-FF prepared in Comparative Example 1.

[0024] Figure 3 Dynamic light scattering (DLS) particle size distribution diagram of AS@DD-FF prepared in Example 1 of the present invention and AS@LL-FF prepared in Comparative Example 1.

[0025] Figure 4 Infrared spectra of AS@DD-FF prepared in Example 1 of the present invention and AS@LL-FF prepared in Comparative Example 1.

[0026] Figure 5 Circular dichroism spectra of AS@DD-FF prepared in Example 1 of the present invention and AS@LL-FF prepared in Comparative Example 1.

[0027] Figure 6 This is a graph showing the toxicity test results of the chiral amino acid-modified arsenene nanomaterials prepared in Comparative Examples 3-6 of the present invention on CT26 cells.

[0028] Figure 7 These are contrasting fluorescence images of live / dead staining of CT26 cells after administration of the unmodified arsenene nanomaterial of the present invention, AS@DD-FF prepared in Example 1, and AS@LL-FF prepared in Comparative Example 1.

[0029] Figure 8Comparison chart of arsenic concentration in CT26 cells after administration of the unmodified arsenene nanomaterial of the present invention, AS@DD-FF prepared in Example 1, and AS@LL-FF prepared in Comparative Example 1.

[0030] Figure 9 Comparison chart of arsenic concentration in the nucleus and organelles of CT26 cells after administration of the unmodified arsenene nanomaterial of the present invention, AS@DD-FF prepared in Example 1, and AS@LL-FF prepared in Comparative Example 1.

[0031] In the figure: A, is the chart of arsenic concentration in the nucleus; B, is the chart of arsenic concentration in the mitochondria; C, is the chart of arsenic concentration in the endoplasmic reticulum; D, is the chart of arsenic concentration in the Golgi apparatus.

[0032] Figure 10 Transmission electron microscope photos of As@LL-FF-fiber prepared in Comparative Example 7 and AS@DD-FF-fiber prepared in Comparative Example 8 of the present invention. Detailed implementation mode

[0033] The present invention will be specifically described below in conjunction with the drawings and embodiments.

[0034] In the present invention, the raw material arsenene nanomaterial used in the following embodiments is an unmodified arsenene nanomaterial, which is synthesized by ultrasonic liquid phase exfoliation method. After ultrasonic treatment in DMSO, differential centrifugation is carried out. The synthesis method of the raw material arsenene nanomaterial is as follows: First, add 500 mg of pure arsenic powder (Aladdin-Aldrich) into 20 mL of DMSO (AladdinAldrich, 99.99% purity) in a plastic centrifuge tube. To avoid the oxidation of arsenene during the sonication process, oxygen in the system is removed by injecting nitrogen for 10 minutes. Then, exfoliation is carried out in an ice bath in a ultrasonic cell disruptor (Ymnl-1800Y) with a power of 1700 W for 120 minutes. Subsequently, the obtained dispersion is washed three times with DMSO to remove the impurities generated during the sonication process. After that, the dispersion is centrifuged at a speed of 6000 revolutions per minute for 2 minutes, and the raw material arsenene nanomaterial is collected.

[0035] Example 1

[0036] A preparation method of a non-natural chiral phenylalanine dipeptide modified arsenene nanomaterial is as follows:

[0037] (1) Disperse 10 mg of D-phenylalanine dipeptide and 6 mg of arsenene nanomaterial in 20 mL of methanol to obtain a dispersion;

[0038] (2) The dispersion was reacted under ultrasonic conditions of 450 W for 30 min. After the reaction was completed, the solvent was removed by rotary evaporation at 40 °C for 1 h under vacuum conditions. After washing with water, centrifugation was carried out at 11,000 rpm for 10 min, and the precipitate was collected to obtain an arsenene nanomaterial modified with unnatural chiral phenylalanine dipeptide AS@DD-FF.

[0039] Example 2

[0040] A preparation method of an arsenene nanomaterial modified with unnatural chiral phenylalanine dipeptide is as follows:

[0041] (1) 5 mg of D-phenylalanine dipeptide and 3 mg of arsenene nanomaterial were dispersed in 20 mL of ethanol to obtain a dispersion.

[0042] (2) The dispersion was reacted under ultrasonic conditions of 450 W for 30 min. After the reaction was completed, the solvent was removed by rotary evaporation at 40 °C for 1 h under vacuum conditions. After washing with water, centrifugation was carried out at 11,000 rpm for 10 min, and the precipitate was collected to obtain an arsenene nanomaterial modified with natural chiral phenylalanine dipeptide AS@DD-FF.

[0043] Example 3

[0044] A preparation method of an arsenene nanomaterial modified with unnatural chiral phenylalanine dipeptide is as follows:

[0045] (1) 10 mg of D-phenylalanine dipeptide and 5 mg of arsenene nanomaterial were dispersed in 20 mL of methanol to obtain a dispersion.

[0046] (2) The dispersion was reacted under ultrasonic conditions of 450 W for 30 min. After the reaction was completed, the solvent was removed by rotary evaporation at 40 °C for 1 h under vacuum conditions. After washing with water, centrifugation was carried out at 11,000 rpm for 10 min, and the precipitate was collected to obtain an arsenene nanomaterial modified with natural chiral phenylalanine dipeptide AS@DD-FF.

[0047] Example 4

[0048] A preparation method of an arsenene nanomaterial modified with unnatural chiral phenylalanine dipeptide is as follows:

[0049] (1) 10 mg of D-phenylalanine dipeptide and 6 mg of arsenene nanomaterial were dispersed in 20 mL of methanol to obtain a dispersion.

[0050] (2) The dispersion was reacted under ultrasonic conditions of 630 W for 25 min. After the reaction was completed, the solvent was removed by rotary evaporation at 36 °C for 0.5 h under vacuum conditions. After washing with water, it was centrifuged at 14000 rpm for 5 min, and the precipitate was collected to obtain the natural chiral phenylalanine dipeptide-modified arsenene nanomaterial AS@DD-FF.

[0051] Comparative Example 1

[0052] A preparation method of a natural chiral phenylalanine dipeptide-modified arsenene nanomaterial is as follows:

[0053] (1) 10 mg of L-phenylalanine dipeptide and 6 mg of arsenene nanomaterial were dispersed in 20 mL of methanol to obtain a dispersion;

[0054] (2) The dispersion was reacted under ultrasonic conditions of 450 W for 30 min. After the reaction was completed, the solvent was removed by rotary evaporation at 40 °C for 1 h under vacuum conditions. After washing with water, it was centrifuged at 11000 rpm for 10 min, and the precipitate was collected to obtain the natural chiral phenylalanine dipeptide-modified arsenene nanomaterial AS@LL-FF.

[0055] Comparative Example 2

[0056] A preparation method of a natural chiral phenylalanine dipeptide-modified arsenene nanomaterial is as follows:

[0057] (1) 5 mg of L-phenylalanine dipeptide and 3 mg of arsenene nanomaterial were dispersed in 20 mL of ethanol to obtain a dispersion;

[0058] (2) The dispersion was reacted under ultrasonic conditions of 450 W for 30 min. After the reaction was completed, the solvent was removed by rotary evaporation at 40 °C for 1 h under vacuum conditions. After washing with water, it was centrifuged at 11000 rpm for 10 min, and the precipitate was collected to obtain the natural chiral phenylalanine dipeptide-modified arsenene nanomaterial AS@LL-FF.

[0059] Comparative Example 3

[0060] A preparation method of an unnatural chiral phenylalanine-modified arsenene nanomaterial is as follows:

[0061] (1) 5 mg of D-phenylalanine and 3 mg of arsenene nanomaterial were dispersed in 20 mL of ultrapure water to obtain a dispersion;

[0062] (2) The dispersion was reacted under ultrasonic conditions of 450 W for 30 min. After the reaction was completed, the solvent was removed by rotary evaporation at 40 °C for 1 h under vacuum conditions. After washing with water, it was centrifuged at 11000 rpm for 10 min, and the precipitate was collected to obtain the unnatural chiral phenylalanine-modified arsenene nanomaterial DPAs.

[0063] Comparative Example 4

[0064] A preparation method of a natural chiral phenylalanine-modified arsenene nanomaterial is as follows:

[0065] (1) Disperse 5 mg of L-phenylalanine and 3 mg of arsenene nanomaterial in 20 mL of ultrapure water to obtain a dispersion;

[0066] (2) Place the dispersion under ultrasonic conditions of 450 W and react for 30 min. After the reaction is completed, under vacuum conditions, rotary evaporate for 1 h at 40 °C to remove the solvent. After washing with water, centrifuge at 11000 rpm for 10 min, collect the precipitate, and obtain the natural chiral phenylalanine-modified arsenene nanomaterial LPAs.

[0067] Comparative Example 5

[0068] A preparation method of an unnatural chiral phenylalanine-modified arsenene nanomaterial is as follows:

[0069] (1) Disperse 5 mg of D-tyrosine and 3 mg of arsenene nanomaterial in 20 mL of ultrapure water to obtain a dispersion;

[0070] (2) Place the dispersion under ultrasonic conditions of 450 W and react for 30 min. After the reaction is completed, under vacuum conditions, rotary evaporate for 1 h at 40 °C to remove the solvent. After washing with water, centrifuge at 11000 rpm for 10 min, collect the precipitate, and obtain the natural chiral phenylalanine dipeptide-modified arsenene nanomaterial DTAs.

[0071] Comparative Example 6

[0072] A preparation method of a natural chiral phenylalanine-modified arsenene nanomaterial is as follows:

[0073] (1) Disperse 5 mg of L-tyrosine and 3 mg of arsenene nanomaterial in 20 mL of ultrapure water to obtain a dispersion;

[0074] (2) Place the dispersion under ultrasonic conditions of 450 W and react for 30 min. After the reaction is completed, under vacuum conditions, rotary evaporate for 1 h at 40 °C to remove the solvent. After washing with water, centrifuge at 11000 rpm for 10 min, collect the precipitate, and obtain the natural chiral phenylalanine dipeptide-modified arsenene nanomaterial LTAs.

[0075] Comparative Example 7

[0076] A preparation method of a natural chiral phenylalanine dipeptide-modified arsenene nanomaterial is as follows:

[0077] (1) Disperse 20 mg of L-phenylalanine dipeptide and 2 mg of arsenene nanomaterials in 20 mL of methanol to obtain a dispersion;

[0078] (2) Place the dispersion under ultrasonic conditions of 450 W and react for 30 min. After the reaction is completed, under vacuum conditions, rotary evaporate for 1 h at 40 °C to remove the solvent. After washing with water, centrifuge at 11000 rpm for 10 min, collect the precipitate, and obtain arsenene nanomaterials modified with natural chiral phenylalanine dipeptide AS@LL-FF-fiber. As Figure 10 shown, this method cannot form sheet-like nanomaterials, but self-assembles into fibers due to the excessive content of phenylalanine dipeptide.

[0079] Comparative Example 8

[0080] A preparation method of arsenene nanomaterials modified with unnatural chiral phenylalanine dipeptide is as follows:

[0081] (1) Disperse 20 mg of D-phenylalanine dipeptide and 2 mg of arsenene nanomaterials in 20 mL of methanol to obtain a dispersion;

[0082] (2) Place the dispersion under ultrasonic conditions of 450 W and react for 30 min. After the reaction is completed, under vacuum conditions, rotary evaporate for 1 h at 40 °C to remove the solvent. After washing with water, centrifuge at 11000 rpm for 10 min, collect the precipitate, and obtain arsenene nanomaterials modified with natural chiral phenylalanine dipeptide AS@DD-FF-fiber. As Figure 10 shown, this method cannot form sheet-like nanomaterials, but self-assembles into fibers due to the excessive content of phenylalanine dipeptide.

[0083] Test Example

[0084] (1) Structural characterization of arsenene nanomaterials

[0085] Perform transmission electron microscopy characterization, atomic force microscopy characterization, dynamic light scattering DLS test, infrared spectroscopy analysis, and circular dichroism analysis on the arsenene nanomaterials prepared in Example 1 and Comparative Example 1. The results are as follows Figures 1 - 5 shown.

[0086] From Figure 1 the results, it can be seen that both the arsenene two-dimensional materials modified with L-phenylalanine dipeptide and the arsenene two-dimensional materials modified with D-phenylalanine dipeptide maintain a large aspect ratio, do not form a tubular structure like traditional self-assembled phenylalanine dipeptide materials, and retain a high surface energy.

[0087] From Figure 2 the results, it can be seen that the thickness of the arsenene two-dimensional materials modified with L-phenylalanine dipeptide and the arsenene two-dimensional materials modified with D-phenylalanine dipeptide is 20 - 30 nm.

[0088] From Figure 3 It can be seen from the results that the widths of both the L-phenylalanine dipeptide modified arsenicene two-dimensional material and the D-phenylalanine dipeptide modified arsenicene two-dimensional material are between 150 - 200 nm.

[0089] From Figure 4 It can be seen from the results that characteristic peaks of LL-FF and DD-FF appear on the L-phenylalanine dipeptide modified arsenicene two-dimensional material and the D-phenylalanine dipeptide modified arsenicene two-dimensional material, confirming the completion of the modification.

[0090] From Figure 5 It can be seen from the results that symmetric spectral peaks appear on the L-phenylalanine dipeptide modified arsenicene two-dimensional material and the D-phenylalanine dipeptide modified arsenicene two-dimensional material, confirming the correspondence of chirality.

[0091] (2) Tumor killing ability test of arsenicene nanomaterials

[0092] Experimental method: Using C33A cells, Bxpc3 cells, Caski cells, Siha cells, CT26 cells, ID8 cells, A549 cells, Pan02 cells, MCF-7 cells as experimental objects, the cells were inoculated into DMEM medium containing 10% FBS and cultured until the cell density reached 100000 cells / mL. Then, for each type of cell, the arsenicene nanomaterials prepared in Example 1 and Comparative Example 1 of the present invention were added at a dose of 5 μg / mL and treated for 12 h and 24 h. After that, the cells were collected and the cell viability was detected by the CCK-8 method. The results are shown in Table 1.

[0093] Table 1 IC50 values of phenylalanine dipeptide modified arsenic nanomaterials for 12 - 24 h

[0094]

[0095] It can be seen from the results in Table 1 that in mouse colon cancer cells (CT26 cells), regardless of whether the treatment time is 12 h or 24 h, the toxicity of As@DD-FF is greater than that of As@LL-FF, and there is a significant difference. In addition, the IC50 value of As@DD-FF treating CT26 cells for 24 h is 1.07 ± 0.02 μg / mL, which is 0.75 times that of As@LL-FF treating CT26 cells for 24 h (1.42 ± 0.07 μg / mL).

[0096] Using CT26 cells as the experimental subject, the cells were inoculated into DMEM medium containing 10% FBS and cultured until the cell density reached 100,000 cells / mL. Then, the chiral amino acid-modified arsenene nanomaterials and unmodified arsenene nanomaterials (control) prepared in Comparative Examples 3-6 of the present invention were added at doses of 0-8 μg / mL and treated for 24 h. The cells were collected, and the cell viability and the half-lethal concentration of the drug were detected by the CCK-8 method. The results are as Figure 6 shown. As can be seen from Figure 6 the results, the killing ability of the chiral amino acid-modified arsenene nanomaterials prepared in Comparative Examples 3-6 against CT26 cells was not improved compared with that of the unmodified arsenene nanomaterials.

[0097] (3) Cytotoxicity test of arsenene nanomaterials

[0098] Experimental method: Using CT26 cells as the experimental subject, the cells were inoculated into the medium of DMEM containing 10% FBS and cultured until the cell density reached 100,000 cells / mL. Then, the arsenene nanomaterials prepared in Example 1 and Comparative Example 1 of the present invention and unmodified arsenene nanomaterials (control) were added at a dose of 5 μg / mL and treated for 24 h. The cells were collected, and the cytotoxicity was detected by staining the cells with calcein AM and propidium iodide respectively. The results are as Figure 7 shown. As can be seen from Figure 7 the results, the modification of chiral phenylalanine dipeptide can effectively improve the cytotoxicity of arsenene nanosheets against CT26, proving that the modification of chiral phenylalanine dipeptide can enhance the inhibition of arsenene nanomaterials on cell viability. In addition, the PI staining fluorescence intensity value of the cells treated with As@DD-FF was 1.38 times higher than that of the As@LL-FF group, which confirmed at the cell level that the chiral difference significantly affects the behavioral characteristics of the phenylalanine dipeptide-modified arsenene nanomaterials.

[0099] (4) Uptake effect test of cells on arsenene nanomaterials

[0100] Experimental method: Using CT26 cells as the experimental subject, the cells were inoculated into the medium of DMEM containing 10% FBS and cultured until the cell density reached 100,000 cells / mL. Then, the arsenene nanomaterials prepared in Example 1 and Comparative Example 1 of the present invention were added at a dose of 5 μg / mL to treat the cells. The cell samples were collected once every 2 h, and the arsenic content in the cells was detected by ICP-MS. The results are as Figure 8 shown. As can be seen from Figure 8The results show that chiral phenylalanine dipeptide modification effectively enhances arsenic uptake in CT26 cells, with a significant difference in intracellular arsenic levels 8 hours after administration. As@DD-FF continuously increases intracellular arsenic levels, while arsenene and As@LL-FF groups reach a peak arsenic content 4 hours later and then gradually decrease, demonstrating the longer-lasting uptake-enhancing effect of As@DD-FF.

[0101] (5) Distribution test of arsenene nanomaterials in organelles

[0102] Experimental method: CT26 cells were used as experimental subjects. The cells were inoculated into DMEM medium containing 10% FBS and cultured to a cell density of 100,000 cells / mL. The arsenene nanomaterials prepared in Example 1 and Comparative Example 1 were added at a dose of 5 μg / mL to treat the cells. Cell samples were collected every 2 hours and the distribution of arsenic in the cells was detected by ICP-MS. The results are shown in Figure 2. Figure 9 As shown. Figure 9 The results show that all three arsenic nanomaterials are primarily enriched in the cell nucleus, followed by the mitochondria, with little accumulation in the endoplasmic reticulum and Golgi apparatus. Arsenic concentration in the nucleus reached its peak at 4 hours, with the As@DD-FF group containing 4.38 times more arsenic than the arsenene group and 1.28 times more arsenic than the As@LL-FF group. Arsenic concentration in the mitochondria also peaked at 4 hours, with the As@DD-FF group containing 2.94 times more arsenic than the arsenene group and 1.09 times more arsenic than the As@LL-FF group. This demonstrates that the introduction of DD-FF significantly improves nuclear delivery efficiency.

[0103] The above description is merely a preferred embodiment of the present invention, and the present invention is not limited to the above embodiment. It is understood that other improvements and variations directly derived or imagined by those skilled in the art without departing from the spirit and concept of the present invention should be considered to be included in the scope of protection of the present invention.

Claims

1. A preparation method of a non-natural chiral phenylalanine dipeptide-modified arsenene nanomaterial, characterized in that, The preparation method includes the following steps: (1) Disperse the phenylalanine dipeptide and the arsenene nanomaterial in a solvent to obtain a dispersion; (2) Under an ice-water bath, ultrasonically treat the dispersion for reaction. After the reaction is completed, remove the solvent by rotary evaporation, then wash with water and centrifuge to collect the precipitate to obtain the phenylalanine dipeptide-modified arsenene nanomaterial.

2. The preparation method according to claim 1, characterized in that, In step (1), the phenylalanine dipeptide is a D-type phenylalanine dipeptide; the mass ratio of the phenylalanine dipeptide to the arsenene nanosheets is 5-6:

3.

3. The preparation method according to claim 1, characterized in that, In step (1), the solvent is any one of methanol and ethanol; the concentration of the phenylalanine dipeptide in the solvent is 0.2-1 mg / mL.

4. The preparation method according to claim 1, characterized in that, In step (2), the power of the ultrasonic treatment is 450-630 W, and the time is 25-30 min.

5. The preparation method according to claim 1, characterized in that, In step (2), the temperature of the rotary evaporation is 36-40 °C, and the time is 0.5-1 h.

6. The preparation method according to claim 1, wherein In step (2), the rotation speed of the centrifugation is 11000-14000 rpm, and the time is 5-10 min.

7. The preparation method according to claim 1, wherein In step (2), the thickness of the phenylalanine dipeptide-modified arsenene nanomaterial is 20-30 nm.

8. Use of an arsenene nanomaterial modified with unnatural chiral phenylalanine dipeptide prepared by the preparation method according to claim 1, characterized in that, The arsenene nanomaterial is used for preparing a drug for treating malignant tumors.

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