Nano-drug for promoting cupric death of tumor cells and preparation method of nano-drug

Abstract

The invention provides a nano medicine for promoting tumor cell copper death and a preparation method thereof, and the preparation method comprises the following steps: selecting a nano raw material carrier, combining copper ion protein, loading a copper ion chelating agent, carrying out ultrasonic wall breaking processing, carrying out surface decoration on the nano carrier, dispersing copper ions, and carrying out dry-method whole particle preparation. According to the nano-drug, the copper ions are carried by the specific nano-carrier, so that the copper ions can be selectively released in tumor cells, the tumor cells are induced to generate copper-dependent death, cell metabolism disorder can be caused by imbalance of a copper steady state, synthesis of iron-sulfur cluster proteins related to mitochondrial respiration is inhibited, and the tumor cell death rate is increased. According to the present invention, with the application of the polypeptide as the protein lipoic acylation modification upstream regulatory factor, the polypeptide can be directly combined with and induce the heteropolymerization, and the drug has good biocompatibility and good targeting property, and the polypeptide can be used as the protein lipoic acylation modification upstream regulatory factor.

Description

Technical Field

[0001] The invention belongs to the field of tumor cell nanomedicines, and particularly relates to a nanomedicine for promoting copper death of tumor cells and a preparation method thereof. Background Art

[0002] Cancer is a major health problem facing the world today. Although traditional chemotherapy drugs can kill tumor cells, they also cause damage to normal cells and lead to serious side effects. Drugs that selectively kill tumor cells and have little effect on normal cells are very important. Copper ions have multiple physiological functions in cells, but excessive copper ions can cause cell death. Copper is a cofactor of essential enzymes required by the human body and can maintain its homeostasis by acting across concentration gradients. Normally, the copper concentration in cells remains at an extremely low level, and this is also true in cancer cells. Copper is transported through the blood system and transferred to the cell surface, where it will be catalyzed and reduced by membrane surface proteins and have stronger cytotoxicity. The copper ions that enter the cells pass through the outer mitochondrial membrane and the inner mitochondrial membrane in turn and enter the mitochondrial matrix. However, existing nanomedicines have certain side effects and cannot cause death of tumor cells, and it is impossible to process and produce nanomedicines.

[0003] Therefore, in view of the shortcomings of the above scheme in actual production and implementation, it is modified and improved. At the same time, in the spirit and concept of seeking improvement, with the assistance of professional knowledge and experience, and after many ingenuity and experiments, the present invention is created to provide a nano drug that promotes copper cell death in tumor cells and a preparation method thereof, which is used to solve the problem. Summary of the invention

[0004] The present invention provides a nano drug for promoting copper death of tumor cells and a preparation method thereof, which solves the problems that the nano drugs in the prior art have certain side effects, cannot cause death of tumor cells, and cannot realize the processing and production of the nano drugs.

[0005] The technical solution of the present invention is achieved as follows: a nano drug that promotes copper cell death in tumor cells and a preparation method thereof, characterized in that the steps of the nano drug processing and preparation process are as follows: (1) Selection of nanomaterial carriers: Nanomedicine uses chitosan as the basic structural unit as the nanomaterial, and nanomaterials with good biocompatibility and the ability to load copper ions can be selected as carriers; (2) Copper ion protein binding: copper ion protein is combined and copper ions are loaded onto the nanocarrier by physical adsorption; (3) Copper ion chelator loading: The normal function of copper ions depends on the interaction of different types of proteins. Copper ions are transported by the blood system and transferred to the cell surface. Cytoplasmic copper ions will also bind to copper ion chelators to neutralize the cytotoxicity of copper. (4) Ultrasonic wall breaking processing: Ultrasonic wall breaking can effectively disperse macromolecules into small molecules or microparticles, thereby preparing multi-morphological nanoparticles, which can be effectively broken by the force of microbubbles to prepare nano-scale powders; (5) Nanocarrier surface decoration: The surface of the nanocarrier can be functionalized to connect tumor cell-specific recognition molecules to improve the targeting of the drug; (6) Copper ion dispersion: The nanocarriers loaded with copper ions are properly dispersed and stabilized to improve the efficacy of copper-death nanomedicines; (7) Dry granulation preparation: The granules are obtained by drying the drug in a drying oven, and then granulated by a rapid granulator to obtain the final nano-drug; As a preferred embodiment, in step (2), the copper ion protein and the nanomaterial are combined by an ion device to achieve transport by physical adsorption.

[0006] As a preferred embodiment, the copper ion chelator in step (3) can regulate the balance of reactive oxygen species in cells and participate in the regulation of cellular copper homeostasis in multiple ways.

[0007] As a preferred embodiment, the copper ions in step (5) can adsorb tumor cells in the human body, such as connecting to tumor cell-specific recognition molecules, to improve the targeting of the drug.

[0008] As a preferred embodiment, the nanocarrier loaded with copper ions in step (6) is dispersed. Disturbance of copper homeostasis can lead to cell metabolic disorders. After excessive accumulation of copper ions due to ion carriers or transport proteins, protein toxicity stress response is induced, which eventually leads to cell death. After adopting the above technical scheme, the beneficial effects of the present invention are as follows: the nanomedicine carries copper ions through specific nanocarriers, and can selectively release copper ions in tumor cells, thereby inducing copper-dependent death of tumor cells. The imbalance of copper homeostasis will lead to cell metabolic disorders. When copper ions are excessively accumulated due to ion carriers or transport proteins, the synthesis of iron-sulfur cluster proteins related to mitochondrial respiration is inhibited, causing protein toxicity stress response. As an upstream regulatory factor of protein lipoylation modification, it can directly bind to and induce heteromerization. This increase in insolubility leads to cellular protein toxicity stress, and the drug has good biocompatibility and targeting, inducing cell death. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0010] Figure 1 This is a schematic diagram of the preparation process structure of the present invention; DETAILED DESCRIPTION

[0011] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments 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 creative work are within the scope of protection of the present invention. Example

[0012] A nano drug that promotes copper cell death in tumor cells and a preparation method thereof, characterized in that the nano drug processing and preparation process steps are as follows: (1) Selection of nanomaterial carriers: Nanomedicine uses chitosan as the basic structural unit as the nanomaterial, and nanomaterials with good biocompatibility and the ability to load copper ions can be selected as carriers; (2) Copper ion protein binding: copper ion protein is combined and copper ions are loaded onto the nanocarrier by physical adsorption; (3) Copper ion chelator loading: The normal function of copper ions depends on the interaction of different types of proteins. Copper ions are transported by the blood system and transferred to the cell surface. Cytoplasmic copper ions will also bind to copper ion chelators to neutralize the cytotoxicity of copper. (4) Ultrasonic wall breaking processing: Ultrasonic wall breaking can effectively disperse macromolecules into small molecules or microparticles, thereby preparing multi-morphological nanoparticles, which can be effectively broken by the force of microbubbles to prepare nano-scale powders; (5) Nanocarrier surface decoration: The surface of the nanocarrier can be functionalized to connect tumor cell-specific recognition molecules to improve the targeting of the drug; (6) Copper ion dispersion: The nanocarriers loaded with copper ions are properly dispersed and stabilized to improve the efficacy of copper-death nanomedicines; (7) Dry granulation preparation: The granules are obtained by drying the drug in a drying oven, and then granulated by a rapid granulator to obtain the final nano-drug; Furthermore, in step (2), the copper ion protein and the nanomaterial are combined by an ion device to achieve transport by physical adsorption.

[0013] Furthermore, the copper ion chelator in step (3) can regulate the balance of reactive oxygen species in cells and participate in the regulation of cellular copper homeostasis in multiple ways.

[0014] Furthermore, the copper ions in step (5) can adsorb tumor cells in the human body, such as connecting to tumor cell-specific recognition molecules, to improve the targeting of the drug.

[0015] Furthermore, the nanocarrier loaded with copper ions in step (6) disperses the copper homeostasis, which may lead to cell metabolic disorders. After excessive accumulation of copper ions due to ion carriers or transporters, it causes protein toxicity stress response and ultimately leads to cell death. Example

[0016] A nano drug that promotes copper cell death in tumor cells and a preparation method thereof, characterized in that the nano drug processing and preparation process steps are as follows: (1) Selection of nanomaterial carriers: Nanomedicine uses chitosan as the basic structural unit as the nanomaterial, and nanomaterials with good biocompatibility and the ability to load copper ions can be selected as carriers; (2) Copper ion protein binding: copper ion protein is combined and copper ions are loaded onto the nanocarrier by physical adsorption; (3) Copper ion chelator loading: The normal function of copper ions depends on the interaction of different types of proteins. Copper ions are transported by the blood system and transferred to the cell surface. Cytoplasmic copper ions will also bind to copper ion chelators to neutralize the cytotoxicity of copper. (4) Ultrasonic wall breaking processing: Ultrasonic wall breaking can effectively disperse macromolecules into small molecules or microparticles, thereby preparing multi-morphological nanoparticles, which can be effectively broken by the force of microbubbles to prepare nano-scale powders; (5) Copper ion dispersion: The nanocarriers loaded with copper ions are properly dispersed and stabilized to improve the efficacy of copper-death nanomedicines; (6) Dry granulation preparation: The granules are obtained by using a drug drying oven, and then granulated by a rapid granulator to obtain the final nano-drug; Furthermore, in step (2), the copper ion protein and the nanomaterial are combined by an ion device to achieve transport by physical adsorption.

[0017] Furthermore, the copper ion chelator in step (3) can regulate the balance of reactive oxygen species in cells and participate in the regulation of cellular copper homeostasis in multiple ways.

[0018] Furthermore, the nanocarrier loaded with copper ions in step (5) disperses the copper homeostasis, which may lead to cell metabolic disorders. After excessive accumulation of copper ions due to ion carriers or transporters, it causes protein toxicity stress response and ultimately leads to cell death.

[0019] Experimental case: Experimental method: The experimental group adopted the method in comparative example 1, and the control group 1 adopted the method in comparative example 1 (removing the step of surface decoration of the nanocarrier).

[0020] Nanomedicine judgment criteria: nanomedicine activity, nanomedicine efficacy, nanomedicine side effects.

[0021] Comparison of the effects of two groups of different nanomedicines, the results are shown in the table below: The invention can improve the activity, efficacy and hemolytic property of the drug through nano-drug modification, improve the extraction rate of active ingredients, enhance the efficacy, and facilitate storage. The imbalance of copper homeostasis can lead to cell metabolism disorder. When copper ions are excessively accumulated due to ion carriers or transport proteins, the synthesis of iron-sulfur cluster proteins related to mitochondrial respiration is inhibited, protein toxicity stress reaction is caused, and finally cell death is caused. As an upstream regulatory factor of protein lipoylation modification, it participates in regulation and can directly bind to and induce heteropolymerization. The increase of this insolubility leads to cell protein toxicity stress and induces cell death, so that the nano-drug carrier interacts well with tumor cells, realizes targeted transport to tumor cells, thereby realizing endocytosis of the drug carrier by tumor cells, and has the characteristics of no side effects and improved drug carrier capacity.

[0022] In the description of the present invention, it is only for the convenience of describing the present invention and simplifying the description, and it is not intended to indicate or imply that the device or element referred to must have a specific orientation, be constructed and operate in a specific orientation, and therefore cannot be understood as a limitation of the present invention. In the description of the present invention, unless otherwise specified and limited, it should be noted that the terms "installed", "connected" and "connected" should be understood in a broad sense, for example, it can be a mechanical connection or an electrical connection, or it can be the internal communication of two elements, it can be a direct connection, or it can be an indirect connection through an intermediate medium. For ordinary technicians in this field, the specific meanings of the above terms can be understood according to specific circumstances.

[0023] The above are only preferred embodiments of the present invention and are 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 protection scope of the present invention.

Claims

1. A nano drug that promotes copper cell death in tumor cells and a preparation method thereof, characterized in that: The steps of nano drug processing and preparation are as follows: (1) Selection of nanomaterial carriers: Nanomedicine uses chitosan as the basic structural unit as the nanomaterial, and nanomaterials with good biocompatibility and the ability to load copper ions can be selected as carriers; (2) Copper ion protein binding: copper ion protein is combined and copper ions are loaded onto the nanocarrier by physical adsorption; (3) Copper ion chelator loading: The normal function of copper ions depends on the interaction of different types of proteins. Copper ions are transported by the blood system and transferred to the cell surface. Cytoplasmic copper ions will also bind to copper ion chelators to neutralize the cytotoxicity of copper. (4) Ultrasonic wall breaking processing: Ultrasonic wall breaking can effectively disperse macromolecules into small molecules or microparticles, thereby preparing multi-morphological nanoparticles, which can be effectively broken by the force of microbubbles to prepare nano-scale powders; (5) Nanocarrier surface decoration: The surface of the nanocarrier can be functionalized to connect tumor cell-specific recognition molecules to improve the targeting of the drug; (6) Copper ion dispersion: The nanocarriers loaded with copper ions are properly dispersed and stabilized to improve the efficacy of copper-death nanomedicines; (7) Dry granulation preparation: The granules are obtained by a drug drying oven and granulated by a rapid granulator to obtain the final nano-drug.

2. A nano drug for promoting copper death of tumor cells and a preparation method thereof according to claim 1, characterized in that: The step (2) combines the copper ion protein and the nanomaterial through an ion device to achieve transportation in a physical adsorption manner.

3. A nano drug for promoting copper death of tumor cells and a preparation method thereof according to claim 1, characterized in that: The copper ion chelator in step (3) can regulate the balance of active oxygen in cells and participate in the regulation of cellular copper homeostasis in multiple ways.

4. A nano drug for promoting copper cell death of tumor cells and a preparation method thereof according to claim 1, characterized in that: In step (5), the copper ions can adsorb tumor cells in the human body, such as connecting to tumor cell-specific recognition molecules, to improve the targeting of the drug.

5. A nano drug for promoting copper death of tumor cells and a preparation method thereof according to claim 1, characterized in that: The nanocarrier loaded with copper ions in step (6) is used to disperse the copper. Disturbance of copper homeostasis can lead to cell metabolic disorders. Excessive accumulation of copper ions due to ion carriers or transport proteins can cause protein toxicity stress response, ultimately leading to cell death.

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