Self-assembling scaffold adjuvant material and preparation method and application thereof

Abstract

The present application relates to a self-assembling scaffold adjuvant material and a preparation method and application thereof. A self-assembling scaffold adjuvant material, comprising micron-sized crystal fibers / rods, wherein the micron-sized crystal fibers / rods are self-assembled by a casein kinase 2 inhibitor. The present application provides a self-assembling scaffold adjuvant material, comprising micron-sized crystal fibers / rods self-assembled by a casein kinase 2 inhibitor, casein kinase 2 is a key factor in regulating the expression of PD-L1 protein in DC cells, and casein kinase 2 inhibitors can inhibit the catalytic function of casein kinase 2 enzymes. Based on the special chemical structure of casein kinase 2 inhibitors, micron-sized crystal fibers / rods are made by utilizing their self-assembly characteristics, which can significantly reduce the expression of PD-L1 protein in DC cells, and injecting them into the body can further self-assemble to form a scaffold structure, which can reshape immune cells and enhance T cell function, and has great application prospects in tumor immunotherapy.

Description

Technical Field

[0001] The present application relates to the field of medical technology, and in particular to a self-assembling scaffold adjuvant material and a preparation method and application thereof. Background Art

[0002] Cancer (malignant tumors) is the main cause of death in humans. In order to conquer diseases such as cancer, scientists have been trying various therapies. Among them, tumor immunotherapy is a major breakthrough in cancer treatment. This method clears tumor cells by activating the anti-tumor immune response in the body, which can restart and maintain the tumor immune cycle, thereby playing the role of anti-tumor immune response, and can clear and control tumor growth. Tumor vaccines are an important tool for tumor immunotherapy, but the antigens in the vaccine have a poor induction effect on adaptive immunity, and adjuvants are needed to enhance antigen presenting cells, mainly for enhancing the activation of dendritic cells (DC). However, traditional adjuvants rely on innate immune mechanisms, and over-activation of innate immunity induces DC maturation with obvious inflammatory side effects. Therefore, it is urgent to develop anti-tumor immune adjuvants that adopt new immune activation mechanisms.

[0003] Research has found that PD-L1 is a protein expressed on the surface of tumor cells. PD-L1 is also expressed on the surface of DC cells. The upregulation of PD-L1 protein expression on the surface of DC cells during immunotherapy will inhibit the body's immune response and is one of the important reasons for tumor immune escape and treatment resistance. The development of an anti-tumor immune adjuvant that can downregulate PD-L1 expression on the surface of DC cells is a key factor in improving the therapeutic effect of tumor immunotherapy. Summary of the invention

[0004] Based on this, the present application provides a self-assembling scaffold adjuvant material that can downregulate the expression of PD-L1 on the surface of DC cells, and a preparation method and application thereof.

[0005] The present application provides a self-assembling scaffold adjuvant material, comprising micron-sized crystal fibers / rods, wherein the material of the micron-sized crystal fibers / rods is self-assembled from casein kinase 2 inhibitors.

[0006] In some of the embodiments, the micron-sized crystal fibers / rods are also loaded with metal ions.

[0007] In some embodiments, the casein kinase 2 inhibitor is selected from one or more of alkaloid compounds, phenolic compounds and flavonoid compounds.

[0008] In some embodiments, the chemical structure of the casein kinase 2 inhibitor contains one or more of the following core structures:

[0009] .

[0010] In some embodiments, the natural casein kinase 2 inhibitor is selected from one or more of ellagic acid, rhamnosine and isoliquiritigenin.

[0011] In some embodiments, the metal ion includes Mg 2+ 、Zn 2+ , Ca 2+ , Mn 2+ , Cu 2+ 、Co 2+ , Fe 2+ and Fe 3+ One or more of .

[0012] In some embodiments, the mass of the metal ions accounts for 0.1% to 10% of the total mass of the self-assembling scaffold adjuvant material.

[0013] In some of the embodiments, the average length of the micron-sized crystal fibers / rods is 10 μm to 90 μm.

[0014] The present application also provides a method for preparing a self-assembling scaffold adjuvant material as described in any of the above embodiments, comprising the following steps:

[0015] Dissolving a powdered casein kinase 2 inhibitor in a good solvent to form a solution;

[0016] A poor solvent is added to the solution to cause the casein kinase 2 inhibitor to self-assemble to form micron-sized crystal fibers / rods.

[0017] In some embodiments, the micron-sized crystal fiber / rod is also loaded with metal ions, and the preparation method further comprises the following steps:

[0018] dispersing the micron-sized crystal fibers / rods in water to form a dispersion;

[0019] The dispersion is mixed with a salt solution containing metal ions to load the metal ions onto the micron-sized crystal fibers / rods.

[0020] The present application also provides the use of the self-assembling scaffold adjuvant material described in any of the above embodiments in the preparation of an anti-tumor immune adjuvant.

[0021] The present application provides a self-assembling scaffold adjuvant material, including micron-sized crystal fibers / rods self-assembled from casein kinase 2 (CK2) inhibitors. Casein kinase 2 is a key factor in regulating the expression of PD-L1 protein in DC cells, and casein kinase 2 inhibitors can inhibit the catalytic function of casein kinase 2 enzyme, thereby facilitating the downregulation of PD-L1 protein expression. Based on the special chemical structure of casein kinase 2 inhibitors, micron-sized crystal fibers / rods are made by utilizing their self-assembly characteristics, which can significantly downregulate the expression of PD-L1 protein in DC cells. The micron-sized crystal fibers / rods are a good anti-tumor immune adjuvant. When injected into the body, the micron-sized crystal fibers / rods can be further self-assembled to form a scaffold structure, which can reshape immune cells and enhance T cell function. The micron-sized crystal fibers / rods have great application prospects in tumor immunotherapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 Schematic diagram of the preparation process of micron-sized crystal fibers / rods in some embodiments;

[0023] Figure 2 The SEM scanning results of the micron-sized crystal fibers / rods prepared in Example 1 at different magnifications;

[0024] Figure 3 This is a flow cytometry result diagram of the ability of the self-assembled scaffold adjuvant material loaded with different metal ions to down-regulate the expression of PD-L1 protein on the surface of DC cells in Example 1;

[0025] Figure 4 This is a curve diagram showing the inhibition of tumor growth in B16F10 tumor-bearing mice using the self-assembled scaffold adjuvant material loaded with different metal ions in Example 1 as an adjuvant;

[0026] Figure 5 The following are photos of tumors in B16F10 tumor-bearing mice after intervention with the self-assembled scaffold adjuvant material loaded with different metal ions in Example 1 as an adjuvant. DETAILED DESCRIPTION

[0027] For ease of understanding of the present application, the present application will be described more fully below in conjunction with embodiments. However, the present application can be implemented in many different forms and is not limited to the embodiments described in the present application. On the contrary, the purpose of providing these embodiments is to make the understanding of the disclosure of the present application more thorough and comprehensive.

[0028] Unless otherwise defined, all technical and scientific terms used in this application have the same meaning as those commonly understood by those skilled in the art to which this application belongs. The terms used herein in the specification of this application are only for the purpose of describing specific embodiments and are not intended to limit this application.

[0029] The terms "and / or", "or / and", and "and / or" used in this article have a selection range that includes any one of two or more related listed items, and also includes any and all combinations of related listed items, and the arbitrary and all combinations include any combination of two related listed items, any more related listed items, or all related listed items. It should be noted that when at least three items are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it should be understood that in this application, the technical solution undoubtedly includes technical solutions that are all connected by "logical and", and undoubtedly includes technical solutions that are all connected by "logical or". For example, "A and / or B" includes three parallel solutions of A, B and "a combination of A and B". For example, the technical solution of "A, and / or, B, and / or, C, and / or, D" includes any one of A, B, C, and D (that is, the technical solution that is all connected by "logical OR"), and also includes any and all combinations of A, B, C, and D, that is, the combination of any two or any three of A, B, C, and D, and also includes the combination of four of A, B, C, and D (that is, the technical solution that is all connected by "logical AND").

[0030] In the present application, when at least three features are connected by at least two conjunctions selected from "and / or", "or / and", and "and / or", it is equivalent to the expression "having any one or any multiple features", for example, "TA, and / or, TB, and / or, TC, and / or, TD" is equivalent to "having any one or any multiple of the following features: TA, TB, TC and TD".

[0031] In the present application, in "the first aspect", "the second aspect", "the third aspect", "the fourth aspect", "the second solvent", etc., the terms "first", "second", "third", "fourth", etc. are used only for descriptive purposes and cannot be understood as indicating or implying relative importance or quantity, nor can they be understood as implicitly indicating the importance or quantity of the indicated technical features. Moreover, "first", "second", "third", "fourth", etc. only serve the purpose of non-exhaustive enumeration and description, and it should be understood that they do not constitute a closed limitation on quantity.

[0032] As used in this application, the terms "comprising," "including," and "comprising" are synonymous and are inclusive or open-ended and do not exclude additional, unrecited members, elements, or method steps.

[0033] In the present application, the technical features described in an open manner include closed technical solutions composed of the listed features, and also include open technical solutions containing the listed features.

[0034] In the present application, the terms “preferred”, “better”, “more preferred” and “suitable” are only used to describe implementation methods or examples with better effects, and it should be understood that they do not constitute limitations on the scope of protection of the present application.

[0035] In the present application, the terms “further”, “furthermore”, “particularly”, etc. are used for descriptive purposes to indicate differences in content, but should not be construed as limiting the scope of protection of the present application.

[0036] In this application, "optionally", "optional" and "optional" refer to optional, that is, to any one of the two parallel schemes of "yes" or "no". If multiple "options" appear in a technical solution, unless otherwise specified and there is no contradiction or mutual restriction, each "optional" is independent.

[0037] In the present application, when it comes to numerical intervals (i.e., numerical ranges), unless otherwise specified, the optional numerical distribution is considered continuous within the above numerical interval, and includes the two numerical endpoints (i.e., the minimum and maximum values) of the numerical range, and each numerical value between the two numerical endpoints. Unless otherwise specified, when the numerical interval only refers to the integers within the numerical interval, it includes the two endpoint integers of the numerical range, and each integer between the two endpoints. In this article, it is equivalent to directly listing each integer, such as "t is an integer selected from 1 to 10" or "t is any integer from 1 to 10", indicating that t is any integer selected from the integer group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9 and 10. In addition, when multiple ranges are provided to describe features or characteristics, these ranges can be merged. In other words, unless otherwise specified, the ranges disclosed herein should be understood to include any and all sub-ranges included therein. The "numerical value" in the numerical interval can be any quantitative value, such as a number, a percentage, a ratio, etc. "Numerical interval" allows broadly including numerical interval types such as percentage intervals, ratio intervals, and ratio intervals.

[0038] Cancer, also known as malignant tumors, is one of the most difficult diseases to overcome in the current medical field. Scientists have been trying to develop effective treatment options, among which immunotherapy is a major breakthrough. In tumor immunotherapy, vaccines are one of the most critical tools, but the antigens in vaccines have a poor induction effect on adaptive immunity, and adjuvants are needed to enhance antigen-presenting cells. Dendritic cells (DC) are considered to be the APC (Antigen-presenting cells) with the strongest antigen-presenting ability. One of the core points of traditional vaccine design is DC activation, which uses innate immunity to make DC cells mature to improve DC cell antigen-presenting ability, but this mechanism is prone to cause obvious inflammatory side effects. Therefore, the development of adjuvants with new immune activation mechanisms will be a key breakthrough in tumor immunotherapy.

[0039] According to research, the PD-L1 molecule on the surface of DC plays a key immunosuppressive role in tumor immunity: on the one hand, PD-L1 on the surface of DC binds to PD-1 on T cells, inhibiting the function of T cells, promoting the production of exhausted T cells, and allowing tumor cells to escape T cell immune attacks; on the other hand, the expression of PD-L1 on the surface of DC is much higher than the expression of its own co-stimulatory factor CD80, and by binding to its own CD80, it blocks the latter from binding to CD28 on the surface of T cells, inhibiting the activation of T cells. Targeted regulation of the expression of PD-L1 protein in DC cells will help enhance the body's immune response and is a key factor in the efficacy of tumor vaccines.

[0040] Based on this, the present application provides a self-assembling scaffold adjuvant material, including micron-sized crystal fibers / rods, wherein the material of the micron-sized crystal fibers / rods is self-assembled by a casein kinase 2 inhibitor.

[0041] Casein kinase 2 (CK2) is a key molecule that regulates the expression of PD-L1 on the surface of DC cells. By regulating the catalytic function of casein kinase 2 (CK2), it is beneficial to achieve the regulation of the expression of PD-L1 protein, and casein kinase 2 inhibitors can affect the catalytic function of casein kinase 2, thereby facilitating the downregulation of the expression of PD-L1 protein. The self-assembling scaffold adjuvant material provided in some embodiments of the present application uses casein kinase 2 inhibitors as active molecules that inhibit the catalytic function of CK2, and utilizes the self-assembly characteristics of the special structure of casein kinase 2 inhibitors to form micron-scale crystal fibers / rods. After being injected into the body as a scaffold vaccine adjuvant, it can play a lymphoid role, further self-assemble into a scaffold structure at the injection site to form a 3D microenvironment, reshape immune cells, and downregulate the expression of PD-L1 protein in DC cells. The above-mentioned material has a strong anti-tumor immune adjuvant function and can significantly inhibit the growth process of tumors.

[0042] It can be understood that the micron-sized crystal fibers / rods in the present application refer to the crystals having a size of micron and a fiber-like and / or rod-like shape.

[0043] It can be understood that the self-assembly involved in the present application refers to the process in which disordered entities of molecules spontaneously arrange into ordered structures (such as fibers, rods) due to non-covalent forces, such as hydrogen bonds, electrostatic forces, van der Waals forces, π-π interactions, hydrophobic interactions and coordination interactions.

[0044] In some embodiments, the micron-sized crystal fibers / rods are also loaded with metal ions. After loading with metal ions, the self-assembled scaffold adjuvant material can further inhibit the enzyme activity of casein kinase 2, thereby further downregulating the expression of PD-L1 protein, thereby further enhancing the effect of inhibiting tumor growth.

[0045] In some embodiments, the casein kinase 2 inhibitor is selected from one or more of alkaloid compounds, phenolic compounds and flavonoid compounds. The chemical structures of substances such as alkaloid compounds, phenolic compounds and flavonoid compounds generally contain structures such as benzene rings, phenolic hydroxyl groups and amino groups. These special structures are prone to self-assembly reactions. For example, benzene rings have π-π interactions, and hydroxyl groups and amino groups are prone to hydrogen bonds. These factors will generate self-assembly driving forces, causing disordered casein kinase 2 inhibitors to spontaneously aggregate and arrange to form an ordered structure.

[0046] It is understood that the "amino" referred to in the present application refers to a derivative of ammonia, having a structural feature of the formula -N(X)2, wherein each "X" is independently H, substituted or unsubstituted alkyl, substituted or unsubstituted cycloalkyl, substituted or unsubstituted heterocyclyl, etc. Non-limiting types of amino groups include -NH2, -N(alkyl)2, -NH(alkyl), -N(cycloalkyl)2, -NH(cycloalkyl), -N(heterocyclyl), -NH(heterocyclyl), -N(aryl), -NH(aryl), -N(alkyl)(aryl), -N(alkyl)(heterocyclyl), -N(cycloalkyl)(heterocyclyl), -N(aryl)(heteroaryl), -N(alkyl)(heteroaryl), etc.

[0047] Furthermore, the chemical structure of the casein kinase 2 inhibitor contains one or more of the following core structures:

[0048] .

[0049] The above-mentioned mother core structure has groups that are prone to self-assembly reaction and are more likely to form micron-sized crystal fibers / rods through self-assembly aggregation.

[0050] Furthermore, the casein kinase 2 inhibitor is selected from one or more of ellagic acid, rhamnosin and isoliquiritigenin.

[0051] Furthermore, the casein kinase 2 inhibitor is selected from ellagic acid. The chemical structure of ellagic acid is as follows:

[0052] Ellagic acid is a phenolic compound that contains multiple benzene rings and multiple phenolic hydroxyl groups in its structure and has significant self-assembly properties.

[0053] In some embodiments, the metal ion includes Mg 2+ 、Zn 2+ , Ca 2+ , Mn 2+ , Cu 2+ 、Co 2+ , Fe 2+ and Fe 3+The above metal ions have a certain inhibitory effect on the catalytic function of casein kinase 2, and loading them on micron-sized crystal fibers / rods can further enhance the ability to downregulate PD-L1. Further, the metal ions include Mg 2+ 、Zn 2+ , Ca 2+ , Mn 2+ , Cu 2+ Preferably, the metal ion is Mg 2+ .

[0054] In some embodiments, the mass of metal ions accounts for 0.1% to 10% of the total mass of the self-assembling scaffold adjuvant material. If the content of metal ions is too low, the synergistic enhancement of down-regulating PD-L1 is not obvious. If the content of metal ions is too high, it may affect its attachment to the micron-sized crystal fiber / rod and easily fall off from the micron-sized crystal fiber / rod, thereby affecting the structural stability of the self-assembling scaffold adjuvant material.

[0055] In some of the embodiments, the average length of the micron-sized crystal fibers / rods is 10 μm to 90 μm.

[0056] Furthermore, in some embodiments, the micron-sized crystal fibers / rods are in a cubic shape, with an average width of 1 μm to 2 μm and an average height of 1 μm to 2 μm.

[0057] like Figure 1 As shown, the present application also provides a method for preparing a self-assembling scaffold adjuvant material as in any of the above embodiments, comprising the following steps:

[0058] Dissolving a powdered casein kinase 2 inhibitor in a good solvent to form a solution;

[0059] Adding a poor solvent into the solution causes the casein kinase 2 inhibitor to self-assemble into micron-sized crystal fibers / rods.

[0060] It can be understood that a good solvent refers to a solvent with a high solubility for casein kinase 2 inhibitors, and a poor solvent refers to a solvent with a low solubility for casein kinase 2 inhibitors. The powdered casein kinase 2 inhibitor is disordered. After it is dissolved in a good solvent to form a solution, a poor solvent is added to reduce the solubility of the casein kinase 2 inhibitor in the mixed solution. Due to the special chemical structure of the casein kinase 2 inhibitor, the casein kinase 2 inhibitor will spontaneously aggregate and precipitate in the solution, and self-assemble to form an ordered fibrous or rod-like structure, i.e., micron-scale crystal fibers / rods.

[0061] Furthermore, the casein kinase 2 inhibitor is ellagic acid, and its good solvent may be, for example, but not limited to, dimethyl sulfoxide (DMSO), ethanol, methanol and other organic solvents. Furthermore, in order to improve the self-assembly ability of the disordered casein kinase 2 inhibitor distributed in the solvent, poor solvents such as water, buffer salt, weak acidic or weak alkaline aqueous solution may be added to promote the aggregation and precipitation of the casein kinase 2 inhibitor molecules to self-assemble and form micron-sized crystal fibers / rods.

[0062] Furthermore, the type of solvent, pH value and other conditions can be adjusted to induce the casein kinase 2 inhibitor to self-assemble and form micron-sized crystal fibers / rods of specific shapes.

[0063] In some embodiments, the micron-sized crystal fibers / rods are also loaded with metal ions, and the preparation method further comprises the following steps:

[0064] Dispersing micron-sized crystal fibers / rods in water to form a dispersion;

[0065] The dispersion is mixed with a salt solution containing metal ions to load the metal ions onto the micron-sized crystal fibers / rods.

[0066] It is understandable that the effect of metal ion loading on micron-sized crystal fibers / rods can be adjusted by adjusting the amount ratio of the dispersion to the salt solution, pH value, temperature, stirring rate and other parameters. It is understandable that metal ions can be loaded onto micron-sized crystal fibers / rods by physical adsorption (such as gaps, pore sizes), chelation (such as the interaction between metal ions and hydroxyl groups), and the like.

[0067] In some embodiments, after the step of mixing the dispersion with a salt solution containing metal ions to load the metal ions onto the micron-sized fibers / rods, the steps of filtering, washing, and drying may also be included. Further, the filtering method may be, for example, centrifugation. Further, the detergent used for washing may be, for example, water, and the number of washings may be 1 to 3 times. Further, the drying method may be, for example, freeze drying or heating and drying.

[0068] The present application also provides the use of the self-assembling scaffold adjuvant material in any of the above embodiments in the preparation of an anti-tumor immune adjuvant.

[0069] The present application is further described in detail below through specific examples. The following examples are more specific, and it can be understood that in other embodiments, it is not limited thereto. In the following specific examples, the instruments, reagents, and materials involved, unless otherwise specified, are conventional instruments, reagents, and materials already available in the prior art, and can be obtained through regular commercial channels. The experimental methods, detection methods, etc. involved in the following embodiments, unless otherwise specified, are conventional experimental methods and detection methods already available in the prior art.

[0070] Example 1

[0071] A preparation method of a self-assembling scaffold adjuvant material comprises the following steps:

[0072] 1. Preparation of micron-sized crystal fibers / rods:

[0073] The ellagic acid powder was added to anhydrous ethanol to maintain a concentration of 2 mg / mL, ultrasonically dissolved, and magnetically stirred for 5 minutes. Water was added dropwise to an ethanol concentration of 80%, and stirring was continued for 2 hours to allow the ellagic acid to fully self-assemble. Subsequently, centrifugation was performed, and the centrifugal speed was set to 12000 rpm for 15 minutes. After the centrifugation, the precipitate was collected and washed with water to remove the residual ethanol in the precipitate to obtain micron-sized crystal fibers / rods (product labeled CK2i).

[0074] 2. Micron-sized crystal fibers / rods loaded with metal ions:

[0075] (1) The micron-sized crystal fibers / rods obtained in step 1 are uniformly dispersed in water to form a dispersion with a concentration of 2 mg / mL.

[0076] (2) Prepare 1 mol / L metal salt solution of magnesium chloride, copper chloride, calcium chloride, zinc chloride, manganese chloride, etc.

[0077] (3) The dispersion obtained in step (1) and the salt solution obtained in step (2) were mixed at a volume ratio of 1:1, and magnetically stirred at 60°C for 4 hours to allow the metal ions in the salt solution to be fully loaded on the micron-sized crystal fibers / rods. The product was then centrifuged at a speed of 12,000 rpm for 15 minutes. After the centrifugation, the precipitate was collected and washed with water to remove the residual metal salt to obtain a self-assembled scaffold adjuvant material (according to the different metal ions, the obtained products were labeled as CK2i+Mg 2+ CK2i+Cu 2+ CK2i+Ca 2+ CK2i+Zn 2+ CK2i+Mn 2+ ).

[0078] Performance characterization:

[0079] 1. Morphology characterization:

[0080] The morphology of the micron-sized crystal fiber / rod prepared in step 1 of Example 1 was observed by scanning electron microscope (SEM). The SEM scanning results are as follows: Figure 2 As shown. Figure 2It can be seen that ellagic acid forms cubic rod-like materials after self-assembly, with an average length of about 43.3µm, a width and height of about 1.9µm, and a relatively rough surface, which is easy to adsorb metal ions.

[0081] 2. Characterization of metal ion loading effect:

[0082] The content of metal ions in the self-assembling scaffold adjuvant material prepared in step 2 of Example 1 was tested. The test results are shown in Table 1 below.

[0083] Table 1

[0084]

[0085] It can be seen from Table 1 that the micron-sized crystal fibers / rods have a 2+ , Cu 2+ , Ca 2+ 、Zn 2+ , Mn 2+ They all have a certain adsorption loading effect, and the adsorption ratio of different metal ions is different.

[0086] 3. Characterization of the ability of self-assembled scaffold adjuvant materials to downregulate PD-L1 on the surface of DCs

[0087] CK2i without metal ions and Mg adsorbed 2+ , Cu 2+ , Ca 2+ 、Zn 2+ , Mn 2+ Self-assembled scaffold adjuvant material (CK2i+Mg 2+ CK2i+Cu 2+ CK2i+Ca 2+ CK2i+Zn 2+ CK2i+Mn 2+ ) were co-cultured with DC2.4 cells at a drug concentration of 100 µg / mL. After 6 hours, flow cytometry was used to analyze the changes in PD-L1 on the surface of 2.4 cells. The test results are as follows Figure 3 As shown. Figure 3 It can be seen that CK2i without metal ion loading can play a certain role in down-regulating the expression of PD-L1 on the DC surface. After CK2i is loaded with metal ions, it can further enhance the ability to down-regulate PD-L1 on the DC surface. In addition, the ability of self-assembled scaffold adjuvant materials loaded with different metal ions to down-regulate PD-L1 on the DC surface is different. 2+ The self-assembled scaffold adjuvant material has the strongest down-regulation function.

[0088] 4. Characterization of the anti-tumor ability of self-assembled scaffold adjuvant materials as anti-tumor immune adjuvants

[0089] Using B16F10 tumor-bearing mice as a model, repeatedly frozen and thawed whole tumor cells as antigens, CK2i without metal ions, and CK2i loaded with different metal ions (Mg 2+ , Cu 2+ ) of self-assembled scaffold adjuvant material (CK2i+Mg 2+ CK2i+Cu 2+ ) is used as an immune adjuvant to intervene and evaluate the anti-tumor immune function of the adjuvant. The test results are as follows Figure 4 and Figure 5 As shown. Figure 4 and Figure 5 It can be seen that the self-assembled scaffold adjuvant materials loaded with different metal ions have certain anti-tumor immune functions, among which the adjuvant loaded with Mg 2+ The self-assembled scaffold adjuvant material has the most significant effect in inhibiting the tumor growth process.

[0090] The technical features of the above-described embodiments may be arbitrarily combined. To make the description concise, not all possible combinations of the technical features in the above-described embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0091] The above-mentioned embodiments only express several implementation methods of the present invention, and the descriptions thereof are relatively specific and detailed, but they cannot be understood as limiting the scope of the invention patent. It should be pointed out that, for ordinary technicians in this field, several variations and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of this application. Therefore, the protection scope of the patent of this application shall be subject to the attached claims.

Claims

1. A self-assembling scaffold adjuvant material, characterized in that: It comprises a micron-sized crystal rod, wherein the material of the micron-sized crystal rod is formed by self-assembly of a casein kinase 2 inhibitor; the micron-sized crystal rod is also loaded with metal ions; the casein kinase 2 inhibitor is selected from ellagic acid; The metal ions include Mg 2+ 、Zn 2+ , Ca 2+ , Mn 2+ , Cu 2+ One or more of the following: the mass of the metal ions accounts for 0.1% to 10% of the total mass of the self-assembling scaffold adjuvant material.

2. The self-assembling scaffold adjuvant material according to claim 1, characterized in that: The average length of the micron-sized crystal rod is 10 μm to 90 μm.

3. A method for preparing the self-assembling scaffold adjuvant material according to any one of claims 1 to 2, characterized in that: The steps include: Dissolving a powdered casein kinase 2 inhibitor in a good solvent to form a solution; Adding a poor solvent to the solution causes the casein kinase 2 inhibitor to self-assemble to form micron-sized crystal rods; The micron-sized crystal rod is also loaded with metal ions, and the preparation method further comprises the following steps: dispersing the micron-sized crystal rods in water to form a dispersion; The dispersion is mixed with a salt solution containing metal ions, so that the metal ions are loaded on the micron-sized crystal rods.

4. Use of the self-assembling scaffold adjuvant material according to any one of claims 1 to 2 in the preparation of an anti-tumor immune adjuvant, characterized in that: The tumor is melanoma.

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