Ti3C2T x Preparation and application of Rh and Ru loaded bimetallic nanozymes

Abstract

The present invention introduces Ti3C2T x Preparation and application of Rh and Ru loaded bimetallic nanozymes, the preparation method comprising: S21, Ti3C2T x Solid, RhCl 3. 3H2O solution and RuCl3 solution are mixed; S22, the mixture obtained in step S21 is dissolved in polyethylene glycol; S23, the product of S22 is ultrasonically dispersed; S24, the product of S23 is transferred to a reactor and subjected to high temperature treatment; S25, the product of S24 is cooled and transferred to a centrifuge tube for washing and centrifugation; S26, the product of S25 is vacuum dried to obtain RhRu / Ti3C2T x Bimetallic nanozyme. The present invention loads metal Rh and Ru into two-dimensional Ti3C2T x Synthesis of RhRu / Ti3C2T on Nanosheets x Bimetallic nanozyme, the bimetallic nanozyme RhRu / Ti3C2T x By utilizing the high hydrogen peroxide tumor microenvironment in osteosarcoma cells to produce reactive oxygen species, thereby alleviating tumor hypoxia, the technical effect of treating osteosarcoma is achieved. At the same time, it provides a new direction for using bimetallic nanozymes to improve the tumor microenvironment and treat osteosarcoma.

Description

Technical Field

[0001] The present invention belongs to the field of tumor drug preparation, and specifically relates to Ti3C2T x Preparation and application of Rh and Ru loaded bimetallic nanozymes. Background Art

[0002] At present, osteosarcoma is the most common primary bone sarcoma, usually diagnosed in adolescence. It is one of the most threatening malignant tumors with poor prognosis and treatment effect, which seriously affects the normal quality of life of patients. The tumor microenvironment is hypoxic and weakly acidic, and the concentration of hydrogen peroxide is too high. Reactive oxygen mainly includes active superoxide anions, hydroxyl free radicals and singlet oxygen. Increasing the level of reactive oxygen in the tumor environment and enhancing the oxidative stress of cancer cells will lead to tumor cell damage and apoptosis. Therefore, regulating the complex microenvironment of the tumor to generate reactive oxygen-mediated treatments is a more feasible method for treating osteosarcoma, such as photodynamic therapy PDT and sonodynamic therapy SDT.

[0003] Clinically, the main treatments for osteosarcoma are surgical resection, adjuvant radiotherapy and chemotherapy, etc. The treatment effect is not good and the prognosis is uncontrollable. In recent years, some nanocatalytic therapies can be used for the treatment of tumors, which is a new treatment method based on nanozymes. Nanozymes are a type of artificial simulated enzymes that have the unique properties of nanomaterials and catalytic functions. Due to their economy, stability and durability, they are widely used in medicine, chemical industry, environment and other fields. Titanium carbide (Ti3C2Tx) is a good nanocarrier load that can be used to load nanozymes. Chinese patent CN112903763A discloses a Ti3C2T x Preparation method of ammonia gas sensor and its light-assisted application, Ti3C2T x The preparation process of ammonia gas sensor includes synthesizing Ti3C2T x , TiO2-Ti3C2T was synthesized by hydrothermal method x The composite thin film sensor for detecting volatile gas components of meat was prepared by screen printing. The obtained gas sensor can achieve better performance in the application of ammonia detection with the help of ultraviolet light, and is expected to be applied to meat freshness monitoring, realizing the identification and concentration detection of volatile gases in fresh meat, which has important practical significance for ensuring the food safety of meat.

[0004] Chinese patent CN110974978A discloses a nanocatalyst for tumor treatment and its preparation method and application. The nanocatalyst provided includes an erythrocyte membrane and a composite nanozyme and a photosensitizer coated in the erythrocyte membrane; the composite nanozyme includes glucose oxidase and iron nanoparticles wrapped in the inner cavity of the glucose oxidase. The nanocatalyst preferentially accumulates at the target tumor site through targeted biomimetic delivery, and releases the composite nanozyme under near-infrared light irradiation; based on the high glucose uptake and weakly acidic environment at the tumor site, glucose oxidase converts glucose into H2O2, inducing iron nanoparticles to initiate the in situ Fenton reaction, and sequentially catalyzes to produce hydroxyl radicals, inducing oxidative damage to tumor cells, and then killing tumor cells. The nanocatalyst can not only achieve efficient loading of the catalyst, but also effectively prolong the circulation time in the body, and achieve accurate and continuous release at the tumor lesion site, providing a new idea and platform for tumor treatment.

[0005] However, there are many problems in the current preparation of nanozymes, such as complex preparation process and large size. Nanozymes themselves also have many disadvantages, such as being difficult to prepare, having poor water solubility and low biocompatibility. Summary of the invention

[0006] To solve the above problems, we aim to prepare a nanozyme that has good water solubility, small size, uniform dispersion, easy preparation and high in vivo biocompatibility, so as to change the complex tumor microenvironment of osteosarcoma cells, produce reactive oxygen species, alleviate tumor hypoxia, and achieve the effect of treating osteosarcoma.

[0007] In order to achieve the above effect, the present invention provides a Ti3C2T x Preparation and application of Rh and Ru loaded bimetallic nanozymes.

[0008] Ti3C2T x Nanozyme catalyst loaded with Rh and Ru bimetallic materials, which consists of RhRu-Ti3C2T x Composition, the RhRu-Ti3C2T x The preparation materials include Ti3C2T x Nanosheets.

[0009] Ti3C2T x A method for preparing a nanozyme catalyst loaded with Rh and Ru bimetallic materials, comprising:

[0010] S21, Ti3C2T x The solid, RhCl3·3H2O solution and RuCl3 solution were mixed;

[0011] S22, dissolving the mixture obtained in step S21 in polyethylene glycol;

[0012] S23, ultrasonically dispersing the product of S22;

[0013] S24, transferring the product of S23 to a reactor and subjecting it to high temperature treatment;

[0014] S25, cooling the product of S24, and transferring it to a centrifuge tube for washing and centrifugation;

[0015] S26. The product of S25 is vacuum dried to obtain the bimetallic nanozyme RhRu-Ti3C2T x .

[0016] Preferably, in the step S21, Ti3C2T x The material ratio of solid, RhCl3·3H2O solution and RuCl3 solution is 8-12mgTi3C2T x Solid: 3-6 ml of 2 mg / mL RhCl3·3H2O solution: 3-6 ml of 10 mg / mL RuCl3 solution.

[0017] Preferably, in the step S22, the polyethylene glycol and Ti3C2T x The solid material ratio is 2-3 mg Ti3C2T x Solid: 5-10 ml of polyethylene glycol with a number average molecular weight of 200.

[0018] Preferably, in step S23, the ultrasonic time is 1-2 hours.

[0019] Preferably, in step S24, the temperature of the high temperature treatment is 160-200° C.; and the time of the high temperature treatment is 12-16 hours.

[0020] Preferably, in step S25, the washing material is anhydrous ethanol and the number of centrifugation is 3 times.

[0021] Preferably, in step S26, the drying time is 12-24 hours.

[0022] Preferably, it is used to treat osteosarcoma.

[0023] The advantages and effects of the application are as follows:

[0024] 1. The present invention loads metal Rh and Ru into two-dimensional Ti3C2T x Synthesis of bimetallic nanozyme RhRu-Ti3C2T on nanosheets x , bimetallic nanozyme RhRu-Ti3C2T x By utilizing the high hydrogen peroxide tumor microenvironment within osteosarcoma cells, reactive oxygen species are produced, thereby alleviating tumor hypoxia and achieving the technical effect of treating osteosarcoma.

[0025] 2. The bimetallic nanozyme RhRu-Ti3C2T provided by the present invention x It has the advantages of small size and uniform dispersion, and has both catalase-like activity and peroxidase activity, and has good biocompatibility and photothermal properties.

[0026] 3. The bimetallic nanozyme RhRu-Ti3C2T provided by the present invention x As a new type of photosensitizer, it catalyzes the production of reactive oxygen species through chemodynamic and photodynamic therapy in the tumor microenvironment, and enhances the enzymatic activity of nanozymes through photothermal therapy, further promoting the production of reactive oxygen species and alleviating the hypoxia of tumor cells. This invention provides a new research direction for bimetallic nanozymes to utilize the tumor microenvironment to produce reactive oxygen species and synergistically treat osteosarcoma through chemodynamic therapy, photodynamic therapy and photothermal therapy.

[0027] The above description is only an overview of the technical solution of the present application. In order to more clearly understand the technical means of the present application so that it can be implemented in accordance with the contents of the specification, and to make the above and other purposes, features and advantages of the present application more obvious and easy to understand, the following is a detailed description of the preferred embodiments of the present application in conjunction with the accompanying drawings as follows.

[0028] Based on the detailed description of the specific embodiments of the present application in combination with the accompanying drawings below, those skilled in the art will become more aware of the above and other objects, advantages and features of the present application. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the drawings required for use in the embodiments or the prior art descriptions are briefly introduced below. Obviously, the drawings described below are some embodiments of the present application. For ordinary technicians in this field, other drawings can also be obtained based on these drawings without creative work. In all drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, each element or part is not necessarily drawn according to the actual scale.

[0030] Figure 1 The Ti3C2T provided by the present invention x A method for preparing a nanozyme catalyst loaded with Rh and Ru bimetallics;

[0031] Figure 2 The Ti3C2T provided by the present invention x High-resolution transmission electron microscopy image of the nanozyme catalyst solid loaded with Rh and Ru bimetallic;

[0032] Figure 3 The Ti3C2T provided by the present invention xCatalase-like activity detection diagram of nanozyme catalyst loaded with Rh and Ru bimetallic;

[0033] Figure 4 The Ti3C2T provided by the present invention x Peroxidase activity detection diagram of nanozyme catalyst loaded with Rh and Ru bimetallic;

[0034] Figure 5 The Ti3C2T provided by the present invention x Photothermal performance test diagram of nanozyme catalyst loaded with Rh and Ru bimetallic;

[0035] Figure 6 The Ti3C2T provided by the present invention x Biocompatibility diagram of Rh and Ru bimetallic nanozyme catalysts;

[0036] Figure 7 The Ti3C2T provided by the present invention x Nanozyme catalyst loaded with Rh and Ru bimetallic kills 143B tumor cells through chemodynamic therapy, photodynamic therapy and photothermal therapy;

[0037] Figure 8 The Ti3C2T provided by the present invention x A diagram showing the ability of Rh and Ru bimetallic nanozyme catalysts to produce ROS within cells. DETAILED DESCRIPTION

[0038] To make the purpose, technical scheme and advantages of the embodiment of the present application clearer, the technical scheme in the embodiment of the present application will be clearly and completely described below in conjunction with the accompanying drawings in the embodiment of the present application. Obviously, the described embodiment is a part of the embodiment of the present application, rather than all of the embodiments. In the following description, specific details such as specific configuration and components are provided only to help fully understand the embodiments of the present application. Therefore, it should be clear to those skilled in the art that various changes and modifications can be made to the embodiments described herein without departing from the scope and spirit of the present application. In addition, for clarity and brevity, the description of known functions and structures is omitted in the embodiment.

[0039] It should be understood that the references to "one embodiment" or "this embodiment" throughout the specification mean that the specific features, structures, or characteristics associated with the embodiment are included in at least one embodiment of the present application. Therefore, the references to "one embodiment" or "this embodiment" appearing throughout the specification do not necessarily refer to the same embodiment. In addition, these specific features, structures, or characteristics may be combined in any suitable manner in one or more embodiments.

[0040] In addition, the present application may repeat reference numerals and / or letters in different examples. This repetition is for the purpose of simplicity and clarity, and does not in itself indicate the relationship between the various embodiments and / or settings discussed.

[0041] The term "and / or" in this article is only a description of the association relationship of associated objects, indicating that there can be three relationships. For example, A and / or B can mean: A exists alone, B exists alone, and A and B exist at the same time. The term " / and" in this article describes another type of association object relationship, indicating that there can be two relationships. For example, A / and B can mean: A exists alone, and A and B exist alone. In addition, the character " / " in this article generally indicates that the previous and next associated objects are in an "or" relationship.

[0042] The term "at least one" in this article is merely a description of the association relationship of associated objects, indicating that there may be three relationships. For example, at least one of A and B can mean: A exists alone, A and B exist at the same time, and B exists alone.

[0043] It should also be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprises" or any other variations thereof are intended to cover non-exclusive inclusions.

[0044] Example 1

[0045] This example mainly introduces Ti3C2T x Nanozyme catalyst loaded with Rh and Ru bimetallic and preparation method thereof.

[0046] Ti3C2T x Nanozyme catalyst loaded with Rh and Ru bimetallic materials, which consists of RhRu-Ti3C2T x Composition, the RhRu-Ti3C2T x The composition materials include Ti3C2T x Nanosheet dispersion, RhCl3·3H2O and RuCl3 solutions.

[0047] Ti3C2T x For the preparation method of Rh and Ru bimetallic nanozyme catalyst, please refer to Figure 1 , Figure 1 The Ti3C2T provided by the present invention x A method for preparing a nanozyme catalyst loaded with Rh and Ru bimetallic materials, comprising:

[0048] S21, Ti3C2T xThe solid, RhCl3·3H2O solution and RuCl3 solution were mixed;

[0049] S22, dissolving the mixture obtained in step S21 in polyethylene glycol;

[0050] S23, ultrasonically dispersing the product of S22;

[0051] S24, transferring the product of S23 to a reactor and subjecting it to high temperature treatment;

[0052] S25, cooling the product of S24, and transferring it to a centrifuge tube for washing and centrifugation;

[0053] S26. The product of S25 is vacuum dried to obtain the bimetallic nanozyme RhRu-Ti3C2T x .

[0054] Furthermore, in the step S21, the required Ti3C2T x The solid mass is 8-12 mg, the RhCl3·3H2O solution is 3-6 ml, and the RuCl3 solution is 3-6 ml.

[0055] Furthermore, in the step S22, the polyethylene glycol is 20-40 ml.

[0056] Furthermore, in step S23, the ultrasonic time is 1-2 hours.

[0057] Furthermore, in the step S24, the temperature of the high temperature treatment is 160-200°C.

[0058] Furthermore, in the step S24, the high temperature treatment time is 12-16 hours.

[0059] Furthermore, in step S25, the washing material is anhydrous ethanol, and the number of centrifugation is 3 times.

[0060] Furthermore, in the step S26, the drying time is 12-24 hours.

[0061] The present invention loads metal Rh and Ru into two-dimensional Ti3C2T x Synthesis of bimetallic nanozyme RhRu-Ti3C2T on nanosheets x , bimetallic nanozyme RhRu-Ti3C2T x By utilizing the high hydrogen peroxide tumor microenvironment within osteosarcoma cells, reactive oxygen species are produced, thereby alleviating tumor hypoxia and achieving the technical effect of treating osteosarcoma.

[0062] The bimetallic nanozyme RhRu-Ti3C2T provided by the present invention xIt has the advantages of small size and uniform dispersion, and also possesses catalase-like activity and peroxidase activity, and has good biocompatibility and photothermal properties.

[0063] The bimetallic nanozyme RhRu-Ti3C2T provided by the present invention x As a new type of photosensitizer, it catalyzes the production of reactive oxygen species through chemodynamic and photodynamic therapy in the tumor microenvironment, and enhances the enzymatic activity of nanozymes through photothermal therapy, further promoting the production of reactive oxygen species and alleviating the hypoxia of tumor cells. This invention provides a new research direction for bimetallic nanozymes to utilize the tumor microenvironment to produce reactive oxygen species and synergistically treat osteosarcoma through chemodynamic therapy, photodynamic therapy and photothermal therapy.

[0064] Example 2

[0065] Based on the above embodiment 1, this embodiment mainly introduces Ti3C2T x Verification of the effect of nanozyme catalyst loaded with Rh and Ru bimetallics.

[0066] Bimetallic nanozyme RhRu-Ti3C2T x Please refer to the high-resolution transmission electron microscopy images of solids Figure 2 ,The morphology, size and distribution of the nanomaterials were observed by high-resolution transmission electron microscopy (HRTEM), and the results showed that the nanomaterials were uniform and small in size.

[0067] Bimetallic nanozyme RhRu-Ti3C2T x Please refer to the catalase activity assay diagram for Figure 3 , the nanozyme RhRu-Ti3C2T was detected by dissolved oxygen instrument x The catalase-like activity test showed that the nanozyme RhRu-Ti3C2T x The concentration of oxygen produced by the decomposition of hydrogen peroxide in the group was the highest and concentration-dependent, indicating that it had good catalase-like activity.

[0068] Bimetallic nanozyme RhRu-Ti3C2T x Please refer to the peroxidase activity assay diagram Figure 4 , through TMB as OH - The capture probe was used to test the peroxidase activity, and its absorbance at 652 nm was detected by an ELISA reader to evaluate the activity of the nanozyme RhRu-Ti3C2T x Decomposition of hydrogen peroxide to produce OH - The ability of nanozyme RhRu-Ti3C2T x The absorbance value first increased and then remained stable within 600s, proving that OH was generated. - , and nanozyme RhRu-Ti3C2Tx Produces OH - The results show that RhRu-Ti3C2T x Nanozymes have good peroxidase activity.

[0069] Bimetallic nanozyme RhRu-Ti3C2T x Please refer to the photothermal performance test diagram Figure 5 , the nanozyme RhRu-Ti3C2T x The irradiation was performed for 10 min, and the change of solution temperature was recorded in real time to evaluate the nanozyme RhRu-Ti3C2T x The photothermal performance of the nanozyme RhRu-Ti3C2T x It has good photothermal performance and is time-dependent and concentration-dependent.

[0070] Bimetallic nanozyme RhRu-Ti3C2T x Please refer to the biocompatibility chart of Figure 6 By integrating the nanozyme RhRu-Ti3C2T x After co-culture with HUVECs cells and 143B cells for 24 h, the biocompatibility of the material was evaluated by CCK-8 assay, and it was found that the nanozyme RhRu-Ti3C2T x In the experimental range of 0-15ug, the survival rate of HUVECs cells was close to 95%, proving that it had no toxicity to normal cells, but had slight toxicity to 143B cells. At a concentration of 15ug / ml, the survival rate of 143B cells was about 85%.

[0071] Bimetallic nanozyme RhRu-Ti3C2T x Please refer to the effect diagram of killing 143B tumor cells by chemodynamic therapy, photodynamic therapy and photothermal therapy. Figure 7 After the nanomaterials were co-cultured with 143B cells and irradiated with near-infrared light, the nanozyme RhRu-Ti3C2T was detected by CCK-8 assay. x The results of photodynamic and photothermal therapy showed that the killing rate of osteosarcoma cells could reach more than 30% under near-infrared light irradiation, indicating that the use of RhRu-Ti3C2T x Nanozyme photodynamic therapy and photothermal therapy can effectively kill osteosarcoma cells.

[0072] Bimetallic nanozyme RhRu-Ti3C2T x Please refer to the diagram showing the ability of cells to produce ROS Figure 8The nanozyme RhRu-Ti3C2T was determined by DCFH-DA fluorescence probe experiment. x The ability to produce ROS in cells shows that the nanozyme RhRu-Ti3C2T x The green fluorescence produced is relatively the strongest, proving that ROS can be generated well and the oxidative stress in cells can be enhanced.

[0073] The above description is only the preferred embodiment of the present invention, which does not limit the protection scope of the present invention. For those skilled in the art, the present invention can be modified and varied in various ways. Within the spirit and principle of the present invention, any change, modification, replacement, integration and parameter change of these embodiments by conventional substitution or capable of achieving the same function without departing from the principle and spirit of the present invention shall fall within the protection scope of the present invention.

Claims

1. Ti3C2T X The use of a nanozyme catalyst loaded with Rh and Ru bimetallics in the preparation of a drug for treating osteosarcoma is characterized in that: The Ti3C2T X The main components of the nanozyme catalyst loaded with Rh and Ru bimetallic are RhRu-Ti3C2T X , the RhRu-Ti3C2T X The preparation materials include Ti3C2T X Nanosheets, RhCl3·3H2O solution, RuCl3 solution; The Ti3C2T X The preparation method of the nanozyme catalyst loaded with Rh and Ru bimetallic materials comprises the following steps: S21, Ti3C2T X The solid, RhCl3·3H2O solution and RuCl3 solution were mixed; S22, dissolving the mixture obtained in step S21 in polyethylene glycol; S23, ultrasonically dispersing the product of S22; S24, transferring the product of S23 to a reactor and subjecting it to high temperature treatment; S25, cooling the product of S24, and transferring it to a centrifuge tube for washing and centrifugation; S26. The product of S25 is dried under vacuum to obtain the bimetallic nanozyme catalyst RhRu-Ti3C2T X ; In the step S24, the temperature of the high temperature treatment is 160-200°C; the time of the high temperature treatment is 12-16h; In the step S26, the drying time is 12-24 hours.

2. Ti3C2T according to claim 1 X The use of a nanozyme catalyst loaded with Rh and Ru bimetallics in the preparation of a drug for treating osteosarcoma is characterized in that: In the S21 step, Ti3C2T X The material ratio of solid, RhCl3·3H2O solution and RuCl3 solution is 8~12mgTi3C2T X Solid: 3-6 ml of RhCl3·3H2O solution with a concentration of 2 mg / mL: 3-6 ml of RuCl3 solution with a concentration of 10 mg / mL.

3. Ti3C2T according to claim 2 X Application of a nanozyme catalyst loaded with Rh and Ru bimetallics in the preparation of a drug for treating osteosarcoma, characterized in that In the step S22, the polyethylene glycol and Ti3C2T X The solid material ratio is 2-3mgTi3C2T X Solid: 5~10ml polyethylene glycol with a number average molecular weight of 200.

4. Ti3C2T according to claim 3 X Application of a nanozyme catalyst loaded with Rh and Ru bimetallics in the preparation of a drug for treating osteosarcoma, characterized in that In the step S23, the ultrasonic time is 1-2 hours.

5. Ti3C2T according to claim 4 X Application of a nanozyme catalyst loaded with Rh and Ru bimetallics in the preparation of a drug for treating osteosarcoma, characterized in that In the step S25, the washing material is anhydrous ethanol, and the number of centrifugation is 3 times.

Images