Plant polyphenol-based nano self-assembled material, and preparation method and application thereof

Abstract

The application discloses a plant polyphenol-based nano self-assembly material and a preparation method and application thereof. The plant polyphenol-based nano self-assembly material is a rod-shaped nanomaterial self-assembled by plant polyphenols, coenzyme Q10 and a surfactant, and the mass ratio of the plant polyphenols, the coenzyme Q10 and the surfactant is 1-20:5-10:2-10. The surfactant is a non-ionic surfactant containing a fatty acid hydrophobic segment, and includes Tween 20, Tween 80, oleic acid polyethylene glycol 2000 and a fatty alcohol polyoxyethylene ether. The application can provide the plant polyphenol-based nano self-assembly material which can specifically target and deeply penetrate myocardial tissue, efficiently isolate chemotherapeutic drugs and does not affect the antitumor effect, and provides technical support for protecting myocardial cells from damage of the chemotherapeutic drugs.

Description

Technical Field

[0001] This invention relates to the field of biomaterials technology, and in particular to a plant polyphenol-based nano-self-assembled material, its preparation method, and its application. Background Technology

[0002] While chemotherapy drugs exert their anti-tumor effects, they also bring serious chemotherapy-induced cardiotoxicity. Because the heart is continuously exposed to chemotherapy drugs throughout the bloodstream, and adult cardiomyocytes have extremely low regenerative capacity, coupled with the overlap in the death mechanisms of cardiomyocytes and tumor cells under chemotherapy, chemotherapy drugs inevitably cause damage to the heart. This cardiotoxicity manifests as various forms of cardiac dysfunction, such as decreased left ventricular ejection fraction, arrhythmias, and even heart failure, and its severity is positively correlated with the cumulative dose of chemotherapy drugs. Therefore, how to limit or prevent the uptake of chemotherapy drugs by cardiomyocytes without affecting the anti-tumor efficacy of the drugs is a critical clinical problem that urgently needs to be solved.

[0003] To reduce the intracellular uptake of chemotherapy drugs by cardiomyocytes, targeted degradation or isolation of these drugs within the cardiac environment is considered a viable strategy. However, achieving efficient isolation of chemotherapy drugs by materials requires that the materials possess both cardiac-targeted delivery capabilities and the ability to penetrate deep into myocardial tissue. While some existing materials exhibit a certain affinity for the abundant collagen fibers in the heart, overcoming the dense structure of myocardial tissue in its complex physiological environment to achieve rapid and deep penetration remains a major technical challenge. Summary of the Invention

[0004] To address the aforementioned issues, this invention aims to provide a plant polyphenol-based nano-self-assembled material, its preparation method, and its application. This material can specifically target and deeply penetrate myocardial tissue, while also efficiently isolating chemotherapy drugs without affecting anti-tumor efficacy.

[0005] The technical solution of the present invention is as follows: On the one hand, a plant polyphenol-based nano-self-assembled material is provided, wherein the plant polyphenol-based nano-self-assembled material is a rod-shaped nanomaterial self-assembled from plant polyphenols, coenzyme Q10 and surfactant, wherein the mass ratio of plant polyphenols, coenzyme Q10 and surfactant is 1-20:5-10:2-10, and the surfactant is a nonionic surfactant containing a fatty acid hydrophobic segment.

[0006] Preferably, the plant polyphenol is any one or more of tannic acid, gallic acid, ellagic acid, and epigallocatechin gallate.

[0007] Preferably, the surfactant is Tween 20, Tween 80, or polyethylene glycol oleate 2000.

[0008] Preferably, the particle size of the plant polyphenol-based nano-self-assembled material is 80-220 nm.

[0009] Preferably, the self-assembly raw materials also include any one or more of metal salts, CTP peptides, and modified CTP peptides, wherein the mass ratio of the metal salt to the plant polyphenol is 1:4-20, and the mass ratio of the CTP peptide or the modified CTP peptide to the plant polyphenol is 500-10000:1.

[0010] Preferably, the metal salt is any one or more of zinc salt, copper salt, and iron salt, and the modified CTP peptide is lauric acid-polyethylene glycol 600-CTP peptide.

[0011] On the other hand, a method for preparing the plant polyphenol-based nano-self-assembled material according to any one of the above claims is also provided, comprising the following steps: S1: Dissolve plant polyphenols in acetone or water to obtain a plant polyphenol solution; dissolve coenzyme Q10 and surfactant in acetone to obtain a coenzyme Q10 solution and a surfactant solution, respectively. S2: Mix the plant polyphenol solution, coenzyme Q10 solution and surfactant solution until homogeneous to obtain a mixed solution; S3: The plant polyphenols, coenzyme Q10 and surfactant in the mixed solution are self-assembled under ultrasonic conditions, and then acetone is removed by dialysis to obtain the plant polyphenol-based nano self-assembled material.

[0012] Preferably, when the raw materials for self-assembly also include metal salts, the metal salts are dissolved in water to obtain a metal salt solution, and then the mixed solution is added dropwise to the metal salt solution before proceeding to step S3. Preferably, when the self-assembly raw materials also include CTP peptides and / or modified CTP peptides, the CTP peptides and / or modified CTP peptides are dissolved in acetone to obtain a peptide solution, and then the peptide solution is mixed with the plant polyphenol solution, coenzyme Q10 solution and surfactant solution.

[0013] Furthermore, the invention also provides the application of any one of the above-mentioned plant polyphenol-based nano-self-assembled materials in the preparation of chemotherapy drugs.

[0014] The beneficial effects of this invention are: This invention provides a plant polyphenol-based nano-self-assembled material that can specifically target and deeply penetrate myocardial tissue, efficiently isolate chemotherapy drugs without affecting anti-tumor efficacy, and provides technical support for protecting myocardial cells from damage by chemotherapy drugs. Attached Figure Description

[0015] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0016] Figure 1 This is a TEM image of the plant polyphenol-based nano-self-assembled material from Example 1. Figure 2 This is a TEM image of the plant polyphenol-based nano-self-assembled material from Example 2. Figure 3 TEM image of the self-assembled material in Comparative Example 2; Figure 4 TEM image of the self-assembled material in Comparative Example 3; Figure 5 The H9c2 myocardial cells in the blank control group and the experimental group showed a 2-fold IC50 ratio with the chemotherapy drug doxorubicin. 50 Schematic diagram of cell viability test results at different concentrations; Figure 6 This is a schematic diagram showing the penetration distance test results of equal-volume spherical nanoparticles and rod-shaped nanomaterials in heart tissue over 24 hours. Detailed Implementation

[0017] The present invention will be further described below with reference to the accompanying drawings and embodiments. It should be noted that, unless otherwise specified, the embodiments and technical features described in this application can be combined with each other. It should also be pointed out that, unless otherwise indicated, all technical and scientific terms used in this application have the same meaning as commonly understood by one of ordinary skill in the art to which this application pertains. The terms "comprising" or "including" and similar words used in this invention refer to elements or objects preceding the word that encompass the elements or objects listed following the word and their equivalents, without excluding other elements or objects.

[0018] On one hand, the present invention provides a plant polyphenol-based nano-self-assembled material, wherein the plant polyphenol-based nano-self-assembled material is a rod-shaped nanomaterial self-assembled from plant polyphenols, coenzyme Q10 and surfactant, wherein the mass ratio of plant polyphenols, coenzyme Q10 and surfactant is 1-20:5-10:2-10, and the surfactant is a nonionic surfactant containing a fatty acid hydrophobic segment.

[0019] In this invention, the plant polyphenols serve as the main component for the self-assembly of rod-shaped nanomaterials and possess cardiac-targeting and chemotherapy drug-adsorbing properties. Coenzyme Q10, as the main hydrophobic component for the self-assembly of rod-shaped nanomaterials, provides a hydrophobic core for these materials. The surfactant ensures the formation and stability of the rod-shaped nanomaterials. The rod-shaped nanostructure formed by the synergistic effect of these three components allows for better penetration into cardiac tissue, achieving a greater penetration distance than ordinary spherical nanoparticles.

[0020] In one specific embodiment, the plant polyphenol is any one or more of tannic acid, gallic acid, ellagic acid, and epigallocatechin gallate. It should be noted that the plant polyphenols in this embodiment are only preferred commonly used plant polyphenols of the present invention, and other plant polyphenols in the prior art can also be applied to the present invention.

[0021] In one specific embodiment, the surfactant is Tween 20, Tween 80, or polyethylene glycol oleate 2000. It should be noted that in this invention, the surfactant is used to enable the plant polyphenols to self-assemble into stable rod-shaped nanostructures. The surfactant in this embodiment is only a preferred surfactant of this invention; other surfactants in the prior art that can achieve the same effect can also be applied to this invention.

[0022] In one specific embodiment, the particle size of the plant polyphenol-based nano-self-assembled material is 80-220 nm.

[0023] In one specific embodiment, the self-assembly raw materials further include any one or more of metal salts, CTP peptides, and modified CTP peptides, wherein the mass ratio of the metal salt to the plant polyphenol is 1:4-20, and the mass ratio of the CTP peptide or the modified CTP peptide to the plant polyphenol is 500-10000:1.

[0024] In the above embodiments, the addition of metal salts can further stabilize the cross-linking of plant polyphenols through their coordination bonds with plant polyphenols, thereby improving structural stability. The addition of the CTP peptide and / or the modified CTP peptide can further enhance the cardiac targeting function of the plant polyphenol-based nano-self-assembled material. It should be noted that the cardiac targeting effect of CTP targets cardiomyocytes. For materials containing CTP to achieve cardiomyocyte targeting, the material must first penetrate into the myocardial tissue before CTP can exert its cardiomyocyte targeting function. That is, CTP does not have the function of penetrating cardiac tissue. This invention first uses the rod-shaped nanostructure of the plant polyphenol-based nano-self-assembled material to penetrate into the myocardial tissue, and then uses CTP for targeting.

[0025] In one specific embodiment, the metal salt is any one or more of zinc salt, copper salt, and iron salt, and the modified CTP peptide is lauric acid-polyethylene glycol 600-CTP peptide.

[0026] On the other hand, the present invention also provides a method for preparing the plant polyphenol-based nano-self-assembled material according to any one of the above claims, comprising the following steps: S1: Dissolve plant polyphenols in acetone or water to obtain a plant polyphenol solution; dissolve coenzyme Q10 and surfactant in acetone to obtain a coenzyme Q10 solution and a surfactant solution, respectively. S2: Mix the plant polyphenol solution, coenzyme Q10 solution and surfactant solution until homogeneous to obtain a mixed solution; S3: The plant polyphenols, coenzyme Q10 and surfactant in the mixed solution are self-assembled under ultrasonic conditions, and then acetone is removed by dialysis to obtain the plant polyphenol-based nano self-assembled material.

[0027] In one specific embodiment, when the raw materials for self-assembly also include metal salts, the metal salts are dissolved in water to obtain a metal salt solution, and then the mixed solution is added dropwise to the metal salt solution before proceeding to step S3. In one specific embodiment, when the self-assembly raw materials also include CTP peptides and / or modified CTP peptides, the CTP peptides and / or modified CTP peptides are dissolved in acetone to obtain a peptide solution, and then the peptide solution is mixed with the plant polyphenol solution, coenzyme Q10 solution and surfactant solution.

[0028] Furthermore, the present invention also provides the application of the plant polyphenol-based nano-self-assembled material described in any one of the above-mentioned claims in the preparation of chemotherapy drugs.

[0029] In this invention, chemotherapeutic drugs are prepared using the aforementioned plant polyphenol-based nano-self-assembled materials. The plant polyphenols themselves contain pyrogallol or catechol moieties in their molecular structure, enabling them to bind strongly to drug molecules through intermolecular forces such as electrostatic interactions, hydrogen bonds, hydrophobic interactions, and π-π conjugation, thus allowing them to act as drug carriers. Furthermore, they can shield against adverse environments such as reactive oxygen species, ultraviolet radiation, and toxic compounds, thereby isolating chemotherapeutic drugs in cardiac tissue and reducing chemotherapy-induced cardiotoxicity. This invention designs the plant polyphenol-based nano-self-assembled materials into rod-shaped nanostructures, enhancing their cardiac-targeted delivery capability and deep myocardial tissue penetration, overcoming the dense structure of myocardial tissue, and achieving rapid and deep penetration of the material into myocardial tissue, thus enabling highly efficient isolation of chemotherapeutic drugs.

[0030] While existing chemotherapy drugs are effective in treating malignant tumors, they can cause severe damage to the heart, leading to cardiac dysfunction. Current technology lacks a means to effectively protect cardiomyocytes from damage caused by various clinically commonly used chemotherapy drugs. The plant polyphenol-based nano-self-assembled material described in this invention can leverage its structural advantages to rapidly penetrate into myocardial tissue and adsorb chemotherapy drugs, thereby solving the problems of myocardial damage and toxicity caused by existing chemotherapy drugs.

[0031] In cardioprotection strategies, a long-standing technical bottleneck has been how to ensure that protective materials not only target the heart but also overcome physiological barriers to achieve rapid penetration and distribution into the deep layers of myocardial tissue. This invention solves this delivery problem by employing rod-shaped plant polyphenol-based self-assembled nanomaterials. Compared to ordinary spherical nanoparticles, these materials leverage their hydrodynamic advantages to rapidly penetrate deep into myocardial tissue in a shorter time, thus ensuring the material can effectively penetrate the myocardial tissue and exert its effects.

[0032] Traditional cardioprotective strategies often risk interfering with the distribution and metabolism of chemotherapy drugs in the body or reducing their plasma concentration, thereby affecting the anti-tumor efficacy. This invention, by utilizing the structural advantages of a rod-shaped material, allows for rapid penetration deep into myocardial tissue and targeted adsorption of chemotherapy drugs, resolving this contradiction. It effectively reduces cardiotoxicity without interfering with the anti-tumor efficacy, in vivo distribution, or plasma concentration of chemotherapy drugs.

[0033] Example 1 A plant polyphenol-based nano-self-assembled material is prepared through the following steps: (1) Tannic acid, coenzyme Q10 and Tween 20 were dissolved in acetone to obtain a tannic acid solution with a concentration of 100 mg / mL, a coenzyme Q10 solution with a concentration of 50 mg / mL and a Tween 20 solution with a concentration of 10 mg / mL. (2) Add 1% of lauric acid-polyethylene glycol 600-CTP peptide (by mass ratio of Tween 20) to 100 μL of the Tween 20 solution, mix well, and then mix it with 100 μL of tannic acid solution and 100 μL of coenzyme Q10 solution. Vortex for 1 min to obtain a mixed solution. (3) Dissolve zinc sulfate heptahydrate in water to obtain a zinc sulfate heptahydrate mother liquor with a concentration of 10 mg / mL; add 0.5 mL of zinc sulfate heptahydrate mother liquor to 4.2 mL of water and sonicate to obtain a zinc sulfate heptahydrate solution with a concentration of 1 mg / mL. (4) The mixed solution is loaded into a 1.0 mL syringe and added dropwise to the zinc sulfate heptahydrate solution under ultrasonic treatment for 3.0 min. (5) Continue ultrasonic treatment for 15 min to allow the raw materials in the mixed solution to self-assemble, and then dialyze to remove acetone to obtain the plant polyphenol-based nano self-assembled material.

[0034] Example 2 Unlike Example 1, in this example, the concentration of tannic acid solution in step (1) is 33 mg / mL, the concentration of coenzyme Q10 solution is 17.7 mg / mL, and the concentration of Tween 20 solution is 3.3 mg / mL; step (3) is omitted, and in step (4), deionized water is used to replace the zinc sulfate heptahydrate solution.

[0035] Example 3 Unlike Example 1, in step (2) of this example, the Tween 20 solution is directly mixed with the tannic acid solution and the coenzyme Q10 solution, that is, lauric acid polyethylene glycol 600-CTP peptide is not added in step (2).

[0036] Example 4 Unlike Example 1, the plant polyphenol in step (1) of this example is EGCG.

[0037] Example 5 Unlike Example 1, the surfactant in step (1) of this example is Tween 80.

[0038] Example 6 Unlike Example 1, in this example, the solvent for tannic acid in step (1) is water, and a tannic acid mother liquor is obtained; the tannic acid mother liquor is not added in step (2); in step (3), 0.1 mL of the tannic acid mother liquor is added to 4.7 mL of water to obtain a tannic acid solution; in step (4), the zinc sulfate heptahydrate solution is replaced with the tannic acid solution.

[0039] Example 7 Unlike Example 1, the metal salt in step (3) of this example is hydrated ferric chloride.

[0040] Comparative Example 1 Unlike Example 1, the self-assembly material in this comparative example does not include Tween 20.

[0041] Comparative Example 2 Unlike Example 1, the self-assembled material in this comparative example does not include tannic acid.

[0042] Comparative Example 3 Unlike Example 1, this comparative example uses oleic acid instead of tannic acid.

[0043] Test Example 1 The morphology of the self-assembled materials in each embodiment and comparative example was observed using transmission electron microscopy, and some results are shown below. Figure 1-4 As shown. From Figure 1 As can be seen, the plant polyphenol-based nano-self-assembled material described in Example 1 of this invention has a uniformly dispersed rod-shaped structure with a particle size distribution concentrated in the range of 80-220 nm, indicating that it has good colloidal stability.

[0044] from Figure 2 As can be seen, the material obtained by self-assembly of plant polyphenols, coenzyme Q10 and surfactants in Example 2 of the present invention also has a uniformly dispersed rod-shaped structure.

[0045] from Figure 3 It can be seen that the material prepared in Comparative Example 2 without the addition of tannic acid exhibits a spherical nanostructure. From Figure 4 It can be seen that the material prepared by replacing tannic acid with oleic acid in Comparative Example 3 is an amorphous material.

[0046] Test Example 2 Cardiomyocyte viability was tested on the self-assembled materials of each embodiment and comparative example. Specifically: at the cell level (using H9c2 as an example): 5000 H9c2 cardiomyocytes / well were added to a 96-well plate and incubated under standard conditions for 12 h to allow them to adhere; the control group was treated with physiological saline mixed with doxorubicin at a concentration of 2 times the IC50 value; the experimental group was treated with a mixture of self-assembled material and doxorubicin, where the doxorubicin concentration was 2 times the IC50 value and the concentration of self-assembled material was 2-5 times the doxorubicin concentration; after incubation in a CO2 incubator for 24 h, cell viability was tested using CCK8 assay, and some results are shown below. Figure 5 As shown. Figure 5 In the figures, Coenzyme Q10, tannic acid, and Tween 20 are the test results of the raw materials for preparing the plant polyphenol-based nano self-assembled material, the polyphenol-based nanomaterial group is the test result of the plant polyphenol-based nano self-assembled material in Example 1, and Coenzyme Q10 + tannic acid and Coenzyme Q10 + Tween 20 are the test results of Comparative Example 1 and Comparative Example 2.

[0047] from Figure 5 It can be seen that coenzyme Q10 alone, tannic acid alone, coenzyme Q10 + tannic acid, coenzyme Q10 + Tween 20, and Tween 20 alone have virtually no anti-doxorphotoxic effects, while the plant polyphenol-based nano-self-assembled material of the present invention can significantly improve the activity of cardiomyocytes treated with doxorphosorbent.

[0048] Test Example 3 The self-assembled materials of each embodiment and comparative example were tested for their permeability to cardiac tissue. Specifically, isolated mouse hearts were taken, perfused with the test material containing fluorescein, and allowed to be gently shaken for 24 hours. The test results for Example 1, Comparative Example 2, and ordinary spherical nanoparticles (50 nm commercially available green fluorescently labeled polystyrene microspheres) are as follows: Figure 6 As shown. From Figure 6 It can be seen that the rod-shaped nanomaterials of this invention have a longer penetration distance than the spherical nanomaterials.

[0049] It should be noted that the test data above are only part of the test data of the present invention. The plant polyphenol-based nano self-assembled materials of other embodiments of the present invention have similar properties to those of Example 1. They all have similar uniformly dispersed rod-shaped structures and strong inter-myocardial tissue permeability. They can specifically target myocardial tissue and reduce the cardiotoxicity of chemotherapy drugs.

[0050] Furthermore, the above embodiments are only some embodiments of the present invention. The rod-shaped nanostructured plant polyphenol-based nano self-assembled materials obtained by changing the type, amount, time, and other parameters of plant polyphenols and surfactants all have similar properties and excellent myocardial tissue penetration ability.

[0051] The above description is merely a representative embodiment of the present invention and is not intended to limit the present invention in any way. Any embodiment made by those skilled in the art without departing from the scope of the present invention and utilizing the disclosed technical content is an equivalent embodiment of the present invention. Any simple modifications, equivalent changes and modifications made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A plant polyphenol-based nano-self-assembled material, characterized in that, The plant polyphenol-based self-assembled nanomaterial is a rod-shaped nanomaterial self-assembled from plant polyphenols, coenzyme Q10, and surfactant. The mass ratio of plant polyphenols, coenzyme Q10, and surfactant is 1-20:5-10:2-10, and the surfactant is a nonionic surfactant containing a hydrophobic segment of fatty acids.

2. The plant polyphenol-based nano-self-assembled material according to claim 1, characterized in that, The plant polyphenols are any one or more of tannic acid, gallic acid, ellagic acid, and epigallocatechin gallate.

3. The plant polyphenol-based nano-self-assembled material according to claim 1, characterized in that, The surfactants are Tween 20, Tween 80, and oleic acid polyethylene glycol 2000.

4. The plant polyphenol-based nano-self-assembled material according to claim 1, characterized in that, The particle size of the plant polyphenol-based nano-self-assembled material is 80-220 nm.

5. The plant polyphenol-based nano-self-assembled material according to any one of claims 1-4, characterized in that, The self-assembly raw materials also include any one or more of metal salts, CTP peptides, and modified CTP peptides, wherein the mass ratio of the metal salt to the plant polyphenol is 1:4-20, and the mass ratio of the CTP peptide or the modified CTP peptide to the plant polyphenol is 500-10000:

1.

6. The plant polyphenol-based nano-self-assembled material according to claim 5, characterized in that, The metal salt is any one or more of zinc salt, copper salt, and iron salt, and the modified CTP peptide is lauric acid-polyethylene glycol 600-CTP peptide.

7. The method for preparing plant polyphenol-based nano-self-assembled materials according to any one of claims 1-6, characterized in that, Includes the following steps: S1: Dissolve plant polyphenols in acetone or water to obtain a plant polyphenol solution; dissolve coenzyme Q10 and surfactant in acetone to obtain a coenzyme Q10 solution and a surfactant solution, respectively. S2: Mix the plant polyphenol solution, coenzyme Q10 solution and surfactant solution until homogeneous to obtain a mixed solution; S3: The plant polyphenols, coenzyme Q10 and surfactant in the mixed solution are self-assembled under ultrasonic conditions, and then acetone is removed by dialysis to obtain the plant polyphenol-based nano self-assembled material.

8. The method for preparing plant polyphenol-based nano-self-assembled materials according to claim 7, characterized in that, When the raw materials for self-assembly also include metal salts, the metal salts are dissolved in water to obtain a metal salt solution, and then the mixed solution is added dropwise to the metal salt solution, and finally the process proceeds to step S3.

9. The method for preparing plant polyphenol-based nano-self-assembled materials according to claim 7 or 8, characterized in that, When the self-assembly raw materials also include CTP peptides and / or modified CTP peptides, the CTP peptides and / or modified CTP peptides are dissolved in acetone to obtain a peptide solution, and then the peptide solution is mixed with the plant polyphenol solution, coenzyme Q10 solution and surfactant solution.

10. The use of the plant polyphenol-based nano-self-assembled material as described in any one of claims 1-6 in the preparation of chemotherapeutic drugs.

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