Carbon dot composite material with anticancer biological activity and preparation method thereof

Abstract

The invention discloses a carbon dot composite material with anticancer biological activity, and particularly relates to the technical field of carbon dot composite materials, the carbon dot composite material comprises nitrogen ruthenium doped carbon dots and sulfhydrylated lactoferrin, and the molar mass ratio of the nitrogen ruthenium doped carbon dots to the sulfhydrylated lactoferrin is (5-30): 1. In the synthesis process, nitrogen and ruthenium are doped, ruthenium is loaded on an sp2 / sp3 hybrid carbon interface when being doped with carbon dots, Ru / NCDs active carbon dots are formed, nitrogen doping is introduced, the dispersity and stability of ruthenium are enhanced, the agglomeration phenomenon is reduced, the photothermal conversion efficiency of the carbon dot composite material when being applied to a terahertz treatment cabin can be effectively improved, and the application range of the composite material is widened. The nitrogen-ruthenium-doped carbon dots are subjected to surface modification by utilizing sulfhydrylated lactoferrin, the penetrability of the carbon dot composite material to specific cells or tissues can be improved by utilizing the targeting property of the lactoferrin, and the lactoferrin has good biocompatibility, so that the immune response of the carbon dots in a living body can be reduced, and the penetrability of the carbon dots is improved.

Description

Technical Field

[0001] The present invention relates to the technical field of carbon dot composite materials, and more specifically, to a carbon dot composite material with anti-cancer biological activity and a preparation method thereof. Background Art

[0002] For many years, the diagnosis and treatment of cancer have been key areas of research worldwide. Despite continuous progress and breakthroughs in both anti-cancer drugs and methods, cancer remains a major threat to human life and health. Current cancer treatments, primarily surgical resection, radiotherapy, chemotherapy, and immunotherapy, fall short of achieving the desired results. Surgical resection not only causes varying degrees of damage to the body but also often precludes patients whose cancer has already spread. Radiotherapy, chemotherapy, and immunotherapy not only have limited efficacy but also commonly present toxic side effects. Therefore, with the increasing incidence of cancer, there is a need to develop anti-cancer drugs with low or no toxic side effects.

[0003] Research literature reports that carbon dots smaller than a certain size can reach the cell membrane and cytoplasm, and carbon dots of different particle sizes can reach locations within the cell. Carbon dots of different particle sizes can be used to label different parts of the cell. Experimental results using imaging of carbon dots fused with cancer cells indicate that after incubation with cells, carbon dots can successfully enter the cells. Specific surface modifications to the carbon dots allow them to better bind to cancer cells. For example, when carbon dot materials are used in terahertz therapy chambers for cancer treatment, the strong penetrability and non-thermal effects of terahertz waves can be used to activate cell metabolism. Carbon dots absorb terahertz energy, generating a photothermal effect that promotes local temperature increases, accelerates blood circulation and metabolism, and can effectively enhance the effectiveness of cancer treatment.

[0004] However, when existing terahertz treatment chambers are used in conjunction with carbon dots for anti-cancer applications, the carbon dots sometimes have poor penetration and photothermal conversion efficiency, which affects the application of carbon dots in anti-cancer treatment. Summary of the Invention

[0005] In order to overcome the above-mentioned defects of the prior art, the embodiments of the present invention provide a carbon dot composite material with anti-cancer biological activity and a preparation method thereof. The problem to be solved by the present invention is: how to improve the activity of the carbon dot material and enhance the anti-cancer therapeutic effect.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a carbon dot composite material with anti-cancer biological activity, comprising nitrogen-ruthenium-doped carbon dots and thiolated lactoferrin, wherein the molar mass ratio of the nitrogen-ruthenium-doped carbon dots to the thiolated lactoferrin is (5-30):1.

[0007] In a preferred embodiment, the molar mass ratio of the nitrogen-ruthenium-doped carbon dots to the thiolated lactoferrin is (15-20):1.

[0008] In a preferred embodiment, the molar mass ratio of the nitrogen-ruthenium-doped carbon dots to the thiolated lactoferrin is 18:1.

[0009] In a preferred embodiment, the preparation method of the nitrogen-ruthenium-doped carbon dots is:

[0010] S1: Weighing p-phenylenediamine and dissolving it in ethanol solvent, reacting it at 170-190° C. for 10-14 hours, and then removing unreacted raw materials by centrifugation to obtain p-phenylenediamine;

[0011] S2: After mixing nitrogen-doped carbon dots with polyethylene glycol, phosphate buffer and ruthenium chloride were added, and the mixture was reacted at room temperature for 2-4 hours. The mixture was filtered and vacuum-dried to obtain nitrogen-ruthenium-doped carbon dots.

[0012] In a preferred embodiment, the mass volume ratio of p-phenylenediamine to ethanol solvent in step S1 is 1-1.5 g p-phenylenediamine to 100-150 ml ethanol solvent, the added amounts of polyethylene glycol and ruthenium chloride in step S2 are (0.3-0.5) and (0.1-0.3) of the mass of nitrogen-ruthenium-doped carbon dots, and the vacuum drying temperature in step S2 is 40-60°C.

[0013] In a preferred embodiment, the preparation method of the thiolated lactoferrin is:

[0014] S1: Incubate the lactoferrin solution with N-succinylidene-S-acetylthioacetate in dimethyl sulfoxide for 25-35 minutes to generate acetylthiolactoferrin;

[0015] S2: adding deacetylation buffer to acetylthiolactoferrin and incubating at room temperature for 1-3 hours to obtain thiolated lactoferrin.

[0016] In a preferred embodiment, the mass ratio of the lactoferrin solution to N-succinylidene-S-acetylthioacetate in step S1 is 1:(0.2-0.8), and the incubation temperature in step S1 is 20-30°C.

[0017] In a preferred embodiment, the deacetylation buffer in step S2 is a hydroxylamine solution.

[0018] The present invention also provides a method for preparing a carbon dot composite material having anti-cancer biological activity, comprising the following steps:

[0019] Step 1: reacting nitrogen-ruthenium-doped carbon dots with thiolated lactoferrin at room temperature for 20-26 hours according to the above molar mass ratio to obtain a premix;

[0020] Step 2: Immerse the premix obtained in step 1 in a phosphate dialysate with a pH of 7.4, and perform dialyzation purification with magnetic stirring to obtain a carbon dot composite material with anti-cancer biological activity.

[0021] In a preferred embodiment, during the dialysis purification in step 2, the dialysate is changed every 30 minutes for the first two hours, and then every 4-6 hours. The total dialysis time is maintained at 48-72 hours, and the temperature during dialysis is maintained at 3-8°C.

[0022] The technical effects and advantages of the present invention are as follows:

[0023] 1. The carbon dot composite material with anti-cancer biological activity of the present invention is composed of nitrogen-ruthenium-doped carbon dots and thiolated lactoferrin. The carbon dots are synthesized by a solvent thermal method to form a super carbon dot structure, which enhances the light-to-heat conversion efficiency. Nitrogen and ruthenium are doped during the synthesis process. When ruthenium is doped, the carbon dots are loaded on the sp2 / sp3 hybridized carbon interface to form Ru / NCDs activated carbon dots. The introduction of nitrogen doping enhances the dispersibility and stability of ruthenium and reduces agglomeration, which can effectively improve the light-to-heat conversion efficiency of the carbon dot composite material when used in a terahertz therapy chamber. The nitrogen-ruthenium-doped carbon dots are surface-modified with thiolated lactoferrin. The targeting property of lactoferrin can enhance the penetration of the carbon dot composite material into specific cells or tissues. In addition, lactoferrin has good biocompatibility and can reduce the immune response of the carbon dots in the body, thereby improving the penetration and bioavailability of the carbon dots.

[0024] 2. The present invention introduces thiol groups into lactoferrin, which can improve the loading and binding of lactoferrin and nitrogen-ruthenium-doped carbon dots, making the carbon dot composite material more effective in anti-cancer treatment. DETAILED DESCRIPTION

[0025] The following will be combined with the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0026] Example 1:

[0027] The present invention provides a carbon dot composite material with anti-cancer biological activity, comprising nitrogen-ruthenium-doped carbon dots and thiolated lactoferrin, wherein the molar mass ratio of the nitrogen-ruthenium-doped carbon dots to the thiolated lactoferrin is 5:1.

[0028] In a preferred embodiment, the preparation method of the nitrogen-ruthenium-doped carbon dots is:

[0029] S1: p-phenylenediamine is weighed and dissolved in ethanol solvent, reacted at 180° C. for 12 hours, and then unreacted raw materials are removed by centrifugation to obtain p-phenylenediamine;

[0030] S2: After mixing nitrogen-doped carbon dots with polyethylene glycol, phosphate buffer and ruthenium chloride were added, and the mixture was reacted at room temperature for 2-4 hours. The mixture was filtered and vacuum-dried to obtain nitrogen-ruthenium-doped carbon dots.

[0031] In a preferred embodiment, the mass volume ratio of p-phenylenediamine to ethanol solvent in step S1 is 1.3 g p-phenylenediamine to 120 ml ethanol solvent, the added amounts of polyethylene glycol and ruthenium chloride in step S2 are 0.4 and 0.2 of the mass of nitrogen-ruthenium-doped carbon dots, and the vacuum drying temperature in step S2 is 50°C.

[0032] In a preferred embodiment, the preparation method of the thiolated lactoferrin is:

[0033] S1: Incubate lactoferrin solution with N-succinylidene-S-acetylthioacetate in dimethyl sulfoxide for 30 minutes to generate acetylthiolactoferrin;

[0034] S2: adding deacetylation buffer to acetylthiolactoferrin and incubating at room temperature for 2 hours to obtain thiolated lactoferrin.

[0035] In a preferred embodiment, the mass ratio of the lactoferrin solution to N-succinylidene-S-acetylthioacetate in step S1 is 1:0.4, and the incubation temperature in step S1 is 25°C.

[0036] In a preferred embodiment, the deacetylation buffer in step S2 is a hydroxylamine solution.

[0037] The present invention also provides a method for preparing a carbon dot composite material having anti-cancer biological activity, comprising the following steps:

[0038] Step 1: reacting nitrogen-ruthenium-doped carbon dots with thiolated lactoferrin at room temperature for 20-26 hours according to the above molar mass ratio to obtain a premix;

[0039] Step 2: Immerse the premix obtained in step 1 in a phosphate dialysate with a pH of 7.4, and perform dialyzation purification with magnetic stirring to obtain a carbon dot composite material with anti-cancer biological activity.

[0040] In a preferred embodiment, during the dialysis purification in step 2, the dialysate is changed every 30 minutes for the first two hours, and then every 5 hours, the total dialysis time is maintained at 52 hours, and the dialysis temperature is maintained at 5°C.

[0041] Example 2:

[0042] Different from Example 1, the molar mass ratio of the nitrogen-ruthenium-doped carbon dots to the thiolated lactoferrin is 18:1.

[0043] Example 3:

[0044] Different from Examples 1-2, the molar mass ratio of the nitrogen-ruthenium-doped carbon dots to the thiolated lactoferrin is 30:1.

[0045] Comparative Example 1:

[0046] The present invention provides a carbon dot composite material with anti-cancer biological activity, comprising nitrogen-ruthenium-doped carbon dots and lactoferrin, wherein the molar mass ratio of the nitrogen-ruthenium-doped carbon dots to the lactoferrin is 5:1.

[0047] In a preferred embodiment, the preparation method of the nitrogen-ruthenium-doped carbon dots is:

[0048] S1: p-phenylenediamine is weighed and dissolved in ethanol solvent, reacted at 180° C. for 12 hours, and then unreacted raw materials are removed by centrifugation to obtain p-phenylenediamine;

[0049] S2: After mixing nitrogen-doped carbon dots with polyethylene glycol, phosphate buffer and ruthenium chloride were added, and the mixture was reacted at room temperature for 2-4 hours. The mixture was filtered and vacuum-dried to obtain nitrogen-ruthenium-doped carbon dots.

[0050] In a preferred embodiment, the mass volume ratio of p-phenylenediamine to ethanol solvent in step S1 is 1.3 g p-phenylenediamine to 120 ml ethanol solvent, the added amounts of polyethylene glycol and ruthenium chloride in step S2 are 0.4 and 0.2 of the mass of nitrogen-ruthenium-doped carbon dots, and the vacuum drying temperature in step S2 is 50°C.

[0051] The present invention also provides a method for preparing a carbon dot composite material having anti-cancer biological activity, comprising the following steps:

[0052] Step 1: reacting nitrogen-ruthenium-doped carbon dots with lactoferrin at room temperature for 20-26 hours according to the above molar mass ratio to obtain a premix;

[0053] Step 2: Immerse the premix obtained in step 1 in a phosphate dialysate with a pH of 7.4, and perform dialyzation purification with magnetic stirring to obtain a carbon dot composite material with anti-cancer biological activity.

[0054] In a preferred embodiment, during the dialysis purification in step 2, the dialysate is changed every 30 minutes for the first two hours, and then every 5 hours, the total dialysis time is maintained at 52 hours, and the dialysis temperature is maintained at 5°C.

[0055] Comparative Example 2:

[0056] The present invention provides a carbon dot composite material with anti-cancer biological activity, comprising nitrogen-doped carbon dots and thiolated lactoferrin, wherein the molar mass ratio of the nitrogen-doped carbon dots to the thiolated lactoferrin is 5:1.

[0057] In a preferred embodiment, the preparation method of the nitrogen-doped carbon dots is:

[0058] S1: p-phenylenediamine is weighed and dissolved in ethanol solvent, reacted at 180° C. for 12 hours, and then unreacted raw materials are removed by centrifugation to obtain p-phenylenediamine;

[0059] S2: Mix the nitrogen-doped carbon dots with polyethylene glycol, add phosphate buffer, react at room temperature for 2-4 hours, filter, and vacuum dry to obtain the nitrogen-doped carbon dots.

[0060] In a preferred embodiment, the mass volume ratio of p-phenylenediamine to ethanol solvent in step S1 is 1.3 g p-phenylenediamine to 120 ml ethanol solvent, the amount of polyethylene glycol added in step S2 is 0.4 of the mass of nitrogen-ruthenium-doped carbon dots, and the vacuum drying temperature in step S2 is 50°C.

[0061] In a preferred embodiment, the preparation method of the thiolated lactoferrin is:

[0062] S1: Incubate lactoferrin solution with N-succinylidene-S-acetylthioacetate in dimethyl sulfoxide for 30 minutes to generate acetylthiolactoferrin;

[0063] S2: adding deacetylation buffer to acetylthiolactoferrin and incubating at room temperature for 2 hours to obtain thiolated lactoferrin.

[0064] In a preferred embodiment, the mass ratio of the lactoferrin solution to N-succinylidene-S-acetylthioacetate in step S1 is 1:0.4, and the incubation temperature in step S1 is 25°C.

[0065] In a preferred embodiment, the deacetylation buffer in step S2 is a hydroxylamine solution.

[0066] The present invention also provides a method for preparing a carbon dot composite material having anti-cancer biological activity, comprising the following steps:

[0067] Step 1: reacting nitrogen-doped carbon dots with thiolated lactoferrin at room temperature for 20-26 hours according to the above molar mass ratio to obtain a premix;

[0068] Step 2: Immerse the premix obtained in step 1 in a phosphate dialysate with a pH of 7.4, and perform dialyzation purification with magnetic stirring to obtain a carbon dot composite material with anti-cancer biological activity.

[0069] In a preferred embodiment, during the dialysis purification in step 2, the dialysate is changed every 30 minutes for the first two hours, and then every 5 hours, the total dialysis time is maintained at 52 hours, and the dialysis temperature is maintained at 5°C.

[0070] Comparative Example 3:

[0071] The present invention provides a carbon dot composite material with anti-cancer biological activity, comprising nitrogen-doped carbon dots and lactoferrin, wherein the molar mass ratio of the nitrogen-doped carbon dots to the lactoferrin is 5:1.

[0072] In a preferred embodiment, the preparation method of the nitrogen-doped carbon dots is:

[0073] S1: p-phenylenediamine is weighed and dissolved in ethanol solvent, reacted at 180° C. for 12 hours, and then unreacted raw materials are removed by centrifugation to obtain p-phenylenediamine;

[0074] S2: Mix nitrogen-doped carbon dots with polyethylene glycol, add phosphate buffer, react at room temperature for 2-4 hours, filter, and vacuum dry to obtain nitrogen-ruthenium-doped carbon dots.

[0075] In a preferred embodiment, the mass volume ratio of p-phenylenediamine to ethanol solvent in step S1 is 1.3 g p-phenylenediamine to 120 ml ethanol solvent, the amount of polyethylene glycol added in step S2 is 0.4 of the mass of nitrogen-ruthenium-doped carbon dots, and the vacuum drying temperature in step S2 is 50°C.

[0076] The present invention also provides a method for preparing a carbon dot composite material having anti-cancer biological activity, comprising the following steps:

[0077] Step 1: reacting nitrogen-doped carbon dots with lactoferrin at room temperature for 20-26 hours according to the above molar mass ratio to obtain a premix;

[0078] Step 2: Immerse the premix obtained in step 1 in a phosphate dialysate with a pH of 7.4, and perform dialyzation purification with magnetic stirring to obtain a carbon dot composite material with anti-cancer biological activity.

[0079] In a preferred embodiment, during the dialysis purification in step 2, the dialysate is changed every 30 minutes for the first two hours, and then every 5 hours, the total dialysis time is maintained at 52 hours, and the dialysis temperature is maintained at 5°C.

[0080] Comparative Example 4:

[0081] The invention comprises nitrogen-ruthenium-doped carbon dots and thiolated lactoferrin, wherein the molar mass ratio of the nitrogen-ruthenium-doped carbon dots to the thiolated lactoferrin is 5:1.

[0082] In a preferred embodiment, the preparation method of the nitrogen-ruthenium-doped carbon dots is:

[0083] S1: p-phenylenediamine is weighed and dissolved in ethanol solvent, reacted at 180° C. for 12 hours, and then unreacted raw materials are removed by centrifugation to obtain p-phenylenediamine;

[0084] S2: After mixing nitrogen-doped carbon dots with polyethylene glycol, phosphate buffer and ruthenium chloride were added, and the mixture was reacted at room temperature for 2-4 hours. The mixture was filtered and vacuum-dried to obtain nitrogen-ruthenium-doped carbon dots.

[0085] In a preferred embodiment, the mass volume ratio of p-phenylenediamine to ethanol solvent in step S1 is 1.3 g p-phenylenediamine to 120 ml ethanol solvent, the added amounts of polyethylene glycol and ruthenium chloride in step S2 are 0.4 and 0.2 of the mass of nitrogen-ruthenium-doped carbon dots, and the vacuum drying temperature in step S2 is 50°C.

[0086] In a preferred embodiment, the preparation method of the thiolated lactoferrin is:

[0087] S1: Incubate lactoferrin solution with N-succinylidene-S-acetylthioacetate in dimethyl sulfoxide for 30 minutes to generate acetylthiolactoferrin;

[0088] S2: adding deacetylation buffer to acetylthiolactoferrin and incubating at room temperature for 2 hours to obtain thiolated lactoferrin.

[0089] In a preferred embodiment, the mass ratio of the lactoferrin solution to N-succinylidene-S-acetylthioacetate in step S1 is 1:0.4, and the incubation temperature in step S1 is 25°C.

[0090] In a preferred embodiment, the deacetylation buffer in step S2 is a hydroxylamine solution.

[0091] The present invention also provides a method for preparing a carbon dot composite material having anti-cancer biological activity, comprising the following steps:

[0092] According to the above molar mass ratio, nitrogen-ruthenium-doped carbon dots and thiolated lactoferrin are reacted at room temperature for 20-26 hours to obtain a carbon dot composite material with anti-cancer biological activity.

[0093] The carbon dot composite materials with anticancer biological activity produced in Examples 1-3 were selected as experimental group 1, experimental group 2, and experimental group 3, respectively, and the carbon dot composite materials produced in Comparative Examples 1-4 were selected as control group 1, control group 2, control group 3, and control group 4, respectively. The photothermal conversion efficiency and the penetrability of the selected carbon dot composite materials were tested in combination with a terahertz treatment chamber;

[0094] Photothermal conversion efficiency: The selected carbon dot composite material was combined with a terahertz therapy chamber to irradiate the experimental subjects under the same conditions. The absorption capacity of the UV-Vis-NIR absorption spectrum was analyzed by spectral analysis, and the temperature was measured 20 minutes after irradiation.

[0095] Penetration detection: The penetration depth of carbon dot composite materials was detected by combining carbon dot composite materials with terahertz treatment chambers of the same size in a rabbit skin model under the same penetration conditions. The fluorescence signal in the in vivo imaging was used to detect the penetration depth of carbon dot composite materials.

[0096] The measurement results are shown in Table 1:

[0097]

[0098] Table 1

[0099] As can be seen from Table 1, when the carbon dot composite material terahertz treatment chamber produced in this embodiment is used for anti-cancer treatment, its photothermal conversion efficiency and penetration effect are good, while the comparative example 1 uses ordinary lactoferrin, and its penetration effect is reduced. The comparative example 2 is not doped with ruthenium, and its photothermal conversion efficiency is reduced. The comparative example 4 has not undergone dialysis purification treatment, and its photothermal conversion efficiency and penetration are slightly reduced. The invention adopts nitrogen-ruthenium-doped carbon dots and thiolated lactoferrin. The carbon dots are synthesized by solvent thermal method to form a super carbon dot structure and enhance the photothermal conversion efficiency. Nitrogen and ruthenium are doped in the synthesis process. When ruthenium is doped with carbon dots, it is loaded on the sp2 / sp3 hybridized carbon interface to form Ru / NCDs active carbon dots. Nitrogen doping is introduced to enhance the dispersion and stability of ruthenium and reduce agglomeration. It can effectively improve the photothermal conversion efficiency of carbon dot composite materials when used in terahertz treatment chambers. Nitrogen-ruthenium-doped carbon dots are surface-modified with thiolated lactoferrin. The targeting property of lactoferrin can improve the penetration of carbon dot composite materials into specific cells or tissues. Lactoferrin has good biocompatibility and can reduce the immune response of carbon dots in the body, thereby improving the penetration and bioavailability of carbon dots. The introduction of thiol groups in lactoferrin can enhance the loading and binding of lactoferrin with nitrogen-ruthenium-doped carbon dots, making the carbon dot composite materials have better anti-cancer effects.

[0100] Finally: The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A carbon dot composite material with anticancer biological activity, characterized in that: The invention comprises nitrogen-ruthenium-doped carbon dots and thiolated lactoferrin, wherein the molar mass ratio of the nitrogen-ruthenium-doped carbon dots to the thiolated lactoferrin is (5-30):

1.

2. The carbon dot composite material with anticancer biological activity according to claim 1, characterized in that: The molar mass ratio of the nitrogen-ruthenium-doped carbon dots to the thiolated lactoferrin is (15-20):

1.

3. The carbon dot composite material with anticancer biological activity according to claim 1, characterized in that: The molar mass ratio of the nitrogen-ruthenium-doped carbon dots to the thiolated lactoferrin is 18:

1.

4. The carbon dot composite material with anticancer biological activity according to claim 1, characterized in that: The preparation method of the nitrogen-ruthenium-doped carbon dots is as follows: S1: Weighing p-phenylenediamine and dissolving it in ethanol solvent, reacting it at 170-190° C. for 10-14 hours, and then removing unreacted raw materials by centrifugation to obtain p-phenylenediamine; S2: After mixing nitrogen-doped carbon dots with polyethylene glycol, phosphate buffer and ruthenium chloride were added, and the mixture was reacted at room temperature for 2-4 hours. The mixture was filtered and vacuum-dried to obtain nitrogen-ruthenium-doped carbon dots.

5. The carbon dot composite material with anti-cancer biological activity according to claim 4, characterized in that: The mass volume ratio of p-phenylenediamine to ethanol solvent in step S1 is 1-1.5 g p-phenylenediamine to 100-150 ml ethanol solvent, the added amounts of polyethylene glycol and ruthenium chloride in step S2 are (0.3-0.5) and (0.1-0.3) of the mass of nitrogen-ruthenium-doped carbon dots, and the vacuum drying temperature in step S2 is 40-60°C.

6. The carbon dot composite material with anti-cancer biological activity according to claim 1, characterized in that: The preparation method of the thiolated lactoferrin is: S1: Incubate the lactoferrin solution with N-succinylidene-S-acetylthioacetate in dimethyl sulfoxide for 25-35 minutes to generate acetylthiolactoferrin; S2: adding deacetylation buffer to acetylthiolactoferrin and incubating at room temperature for 1-3 hours to obtain thiolated lactoferrin.

7. The carbon dot composite material with anticancer biological activity according to claim 6, characterized in that: The mass ratio of the lactoferrin solution to N-succinylidene-S-acetylthioacetate in step S1 is 1:(0.2-0.8), and the incubation temperature in step S1 is 20-30°C.

8. The carbon dot composite material with anti-cancer biological activity according to claim 6, characterized in that: In step S2, the deacetylation buffer is a hydroxylamine solution.

9. The method for preparing a carbon dot composite material with anticancer biological activity according to any one of claims 1 to 8, characterized in that: The following steps are involved: Step 1: reacting nitrogen-ruthenium-doped carbon dots with thiolated lactoferrin at room temperature for 20-26 hours according to the above molar mass ratio to obtain a premix; Step 2: Immerse the premix obtained in step 1 in a phosphate dialysate with a pH of 7.4, and perform dialyzation purification with magnetic stirring to obtain a carbon dot composite material with anti-cancer biological activity.

10. The method for preparing a carbon dot composite material with anti-cancer biological activity according to claim 9, characterized in that: During the dialysis purification in step 2, the dialysis fluid is changed every 30 minutes for the first two hours, and then every 4-6 hours. The total dialysis time is maintained at 48-72 hours, and the temperature during dialysis is maintained at 3-8°C.

Images