Co-assembled fibrous drug carrier as well as preparation method and application thereof
Through the preparation method of co-assembled fibrous drug carriers, the problem of low solubility and bioavailability of flavonoid drugs such as quercetin in clinical applications is solved, and efficient drug delivery and anti-inflammatory effects are achieved, which is suitable for long-term treatment.
Patent Information
- Application Number
- CN202411415987.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-11
- Publication Date
- 2025-01-07
AI Technical Summary
Flavonoid drugs such as quercetin are restricted by problems such as poor water solubility, low bioavailability and short half-life in clinical applications. Especially when long-term treatment is required, it is difficult to achieve effective inflammatory treatment.
By using the preparation method of co-assembled fibrous drug carrier, fibers are formed by dissolving quercetin in anhydrous ethanol and tannin acid in pure water, stirring, ultrasonic bubble removal, refrigeration and freeze-drying. , and enhance drug delivery ability.
It realizes efficient encapsulation and delivery of drugs, reduces side effects of drugs, improves efficacy and bioavailability, and is suitable for alcohol-soluble drugs, especially in inflammatory treatment, with significant anti-inflammatory effects.
Smart Images
Figure CN119258226A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of medical technology, and in particular to a co-assembled fibrous drug carrier and a preparation method and application thereof. Background Art
[0002] As a natural bioflavonoid, quercetin has a variety of pharmacological effects, including anti-inflammatory, antioxidant, anti-cancer and antimicrobial effects. However, its application is restricted by some limitations. For example, poor water solubility, low bioavailability and short half-life. These factors together lead to the limitations of flavonoids such as quercetin in clinical applications, especially when long-term treatment is required. In order to overcome these problems, this project proposes a method for preparing a new drug carrier that can self-assemble with drugs into fibrous products, improve the solubility and bioavailability of alcohol-soluble drugs such as rutin, quercetin, paclitaxel, etc., and enhance drug delivery capabilities to achieve more effective inflammation treatment. Summary of the invention
[0003] The purpose of the present invention is to design and prepare a co-assembled fibrous drug carrier and a preparation method and application thereof.
[0004] The present invention also provides a method for preparing a co-assembled fibrous drug carrier, which has a simple synthesis. The drug prepared by the method has excellent drug encapsulation rate and drug loading rate, solubility and bioavailability, and enhances drug delivery ability, thereby achieving more effective inflammation treatment. The present invention solves its technical problem by adopting the following technical scheme. The present invention proposes a novel co-assembled fibrous drug carrier, which includes: first dissolving an alcohol-soluble drug in anhydrous ethanol to prepare solution A, and then dissolving tannic acid in pure water to prepare solution B. Solution B is added to solution A and stirred to dissolve, and ultrasonically debubbled, refrigerated and allowed to stand, and a fibrous product is obtained by post-treatment. The above-mentioned mixed solution is centrifuged and washed with pure water, frozen at -80°C, and freeze-dried to obtain a fibrous product. The product is a co-assembled fibrous drug.
[0005] A method for preparing a co-assembled fibrous drug carrier comprises the following steps:
[0006] S1. First, dissolve quercetin in anhydrous ethanol to prepare solution A;
[0007] S2, dissolving tannic acid in water to prepare solution B;
[0008] S3. Add solution B to solution A and stir to dissolve, remove bubbles by ultrasound, refrigerate and let stand, and obtain a fibrous product through post-treatment as a fibrous drug carrier.
[0009] In step S1, the concentration of quercetin in the solution A is 1-6 mg / ml.
[0010] In step S2, the concentration of tannic acid in the solution B is 1-6 mg / ml.
[0011] In step S3, the volume ratio of solution A to solution B is 1:4-10.
[0012] In step S3, the refrigerated storage conditions are: refrigerated storage at 2-6° C. for 2-5 days.
[0013] In step S3, the post-processing includes:
[0014] The mixed solution after refrigeration and standing is centrifuged at 4000-6000 rpm, and the solid product is washed with water and collected, and freeze-dried to obtain a fibrous product. The freeze-drying conditions are: freeze-drying at -85°C to -75°C for 24-48 hours.
[0015] The application of the fibrous drug carrier in the preparation of drugs for treating inflammation.
[0016] Compared with the prior art, the present invention has the following advantages:
[0017] The beneficial effect of the preparation method of a novel co-assembled fibrous drug carrier of an embodiment of the present invention is that the co-assembled fibrous drug prepared by the present invention is not only applicable to various types of alcohol-soluble drugs, but also can improve the bioavailability and targeting of drugs, while minimizing systemic effects. In short, drug carriers can accurately deliver drugs to target tissues or cells by using self-assembly technology, thereby reducing the impact on non-target sites and improving the therapeutic effect. In addition, the prepared drug carrier has an excellent drug loading rate and drug encapsulation rate, which helps to protect the drug from degradation. The preparation method of a novel co-assembled fibrous drug carrier provided by the present invention forms co-assembled fibers through non-covalent interactions such as hydrophilic / hydrophobic interactions and π-π stacking, which can effectively reduce drug side effects, improve efficacy, control drug release, slow down drug degradation, prevent aggregation and chemical reactions between drug molecules, etc. Therefore, this drug carrier strategy has broad prospects for clinical application. BRIEF DESCRIPTION OF THE DRAWINGS
[0018] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments are briefly introduced below. It should be understood that the following drawings only show certain embodiments of the present invention and therefore should not be regarded as limiting the scope. For ordinary technicians in this field, other related drawings can be obtained based on these drawings without creative work.
[0019] Figure 1 The surface images of the fibrous drug carriers provided in Example 1, Example 2, Example 3, Example 4, and Example 5 of the present invention.
[0020] Figure 2 Microscopic images of carrier drugs provided in Example 1, Example 2, Example 3, Example 4, and Example 5 of the present invention.
[0021] Figure 3 This is the X-ray diffraction pattern of tannic acid and quercetin in Example 1 of the present invention.
[0022] Figure 4 This is a statistical graph of drug loading rates provided for Example 1, Example 2, Example 3, Example 4, and Example 5 of the present invention.
[0023] Figure 5 This is a statistical graph of encapsulation efficiency provided for Example 1, Example 2, Example 3, Example 4, and Example 5 of the present invention.
[0024] Figure 6 This is a statistical graph of cell activity provided by Example 1, Example 2, Example 3, Example 4, and Example 5 of the present invention.
[0025] Figure 7 This is a microscopic image provided for Example 6 of the present invention.
[0026] Figure 8 1 is the potential diagram of tannic acid and quercetin according to the present invention.
[0027] Fig. 9 Potential diagrams provided for Example 1, Example 2, Example 3, Example 4, and Example 5 of the present invention.
[0028] Fig.10 This is a statistical chart of the wound healing area of Application Example 1 of the present invention.
[0029] Fig.11 This is the average fluorescence intensity statistics of Application Example 1 of the present invention. DETAILED DESCRIPTION
[0030] In order to make the purpose, technical scheme and advantages of the embodiment of the present invention clearer, the technical scheme in the embodiment of the present invention will be described clearly and completely below. The reagents or instruments used without indicating the manufacturer are all conventional products that can be purchased commercially. The preparation method of the novel drug carrier of the embodiment of the present invention is specifically described below.
[0031] The present invention provides a method for preparing a novel co-assembled fibrous drug carrier, which comprises: dissolving quercetin, a flavonoid Chinese medicine monomer, in 10 ml of anhydrous ethanol and dissolving it completely to prepare a solution A. In a preferred embodiment of the present invention, the concentration of the quercetin is preferably 3 mg / ml.
[0032] A certain amount of tannic acid is dissolved in 40 ml of pure water to prepare solution B. In a preferred embodiment of the present invention, the preferred tannic acid concentration is 1 mg / ml.
[0033] Solution B is added to solution A and stirred to dissolve, and then ultrasonically removed bubbles and refrigerated for a period of time. In a preferred embodiment of the present invention, the preferred volume ratio of solution B to solution A is 4:1, the refrigeration temperature is 4°C, and the refrigeration time is 3 days.
[0034] The mixed solution was centrifuged and the solid product was washed with pure water, frozen at -80°C for 48 hours, and then dried and collected in a freeze dryer.
[0035] The invention provides a novel method for preparing a co-assembled fibrous drug carrier, which has the beneficial effects of having an excellent encapsulation rate and a release rate. It can be used as a drug carrier strategy, is suitable for alcohol-soluble drugs, has a broad spectrum, can effectively reduce drug side effects, improve efficacy, and help protect drugs from degradation.
[0036] The features and performance of the present invention are further described in detail below in conjunction with the embodiments.
[0037] Example 1
[0038] The present invention provides a preparation method of a co-assembled fibrous drug carrier, which comprises: firstly dissolving quercetin, a flavonoid Chinese medicine monomer, in 10 ml of anhydrous ethanol (3 mg / ml), and dissolving it thoroughly to prepare solution A. Dissolving a certain amount of tannic acid (1 mg / ml) in pure water to prepare solution B. Adding solution A dropwise to solution B and stirring to dissolve, the volume ratio of solution A to solution B is 1:4, and ultrasonically removing bubbles, refrigerating at 4°C and standing for 3 days. Centrifuging the above-mentioned mixed solution at 5000 rpm, washing the solid product with pure water and collecting it, freezing it at -80°C for 48 hours, and freeze-drying to obtain a fibrous product.
[0039] Example 2
[0040] The present invention provides a preparation method of a co-assembled fibrous drug carrier, which comprises: firstly dissolving quercetin, a flavonoid Chinese medicine monomer, in 10 ml of anhydrous ethanol (1 mg / ml), and dissolving it thoroughly to prepare solution A. Dissolving a certain amount of tannic acid (3 mg / ml) in pure water to prepare solution B. Adding solution A dropwise to solution B and stirring to dissolve, the volume ratio of solution A to solution B is 1:4, and ultrasonically removing bubbles, refrigerating at 4°C and standing for 3 days. Centrifuging the above-mentioned mixed solution at 5000 rpm, washing the solid product with pure water and collecting it, freezing it at -80°C for 48 hours, and freeze-drying to obtain a fibrous product.
[0041] Example 3
[0042] The present invention provides a preparation method of a co-assembled fibrous drug carrier, which comprises: firstly dissolving quercetin, a flavonoid Chinese medicine monomer, in 10 ml of anhydrous ethanol (2 mg / ml), and dissolving it thoroughly to prepare solution A. Dissolving a certain amount of tannic acid (3 mg / ml) in pure water to prepare solution B. Adding solution A dropwise to solution B and stirring to dissolve, the volume ratio of solution A to solution B is 1:4, and ultrasonically removing bubbles, refrigerating at 4°C and standing for 3 days. Centrifuging the above-mentioned mixed solution at 5000 rpm, washing the solid product with pure water and collecting it, freezing it at -80°C for 48 hours, and freeze-drying to obtain a fibrous product.
[0043] Example 4
[0044] The present invention provides a preparation method of a co-assembled fibrous drug carrier, which comprises: firstly dissolving quercetin, a flavonoid Chinese medicine monomer, in 10 ml of anhydrous ethanol (3 mg / ml), and dissolving it thoroughly to prepare solution A. Dissolving a certain amount of tannic acid (3 mg / ml) in pure water to prepare solution B. Adding solution A dropwise to solution B and stirring to dissolve, the volume ratio of solution A to solution B is 1:4, and ultrasonically removing bubbles, refrigerating at 4°C and standing for 3 days. Centrifuging the above-mentioned mixed solution at 5000 rpm, washing the solid product with pure water and collecting it, freezing it at -80°C for 48 hours, and freeze-drying to obtain a fibrous product.
[0045] Example 5
[0046] The present invention provides a preparation method of a co-assembled fibrous drug carrier, which comprises: firstly dissolving quercetin, a flavonoid Chinese medicine monomer, in 10 ml of anhydrous ethanol (3 mg / ml), and dissolving it thoroughly to prepare solution A. Dissolving a certain amount of tannic acid (2 mg / ml) in pure water to prepare solution B. Adding solution A dropwise to solution B and stirring to dissolve, the volume ratio of solution A to solution B is 1:4, and ultrasonically removing bubbles, refrigerating at 4°C and standing for 3 days. Centrifuging the above-mentioned mixed solution at 5000 rpm, washing the solid product with pure water and collecting it, freezing it at -80°C for 48 hours, and freeze-drying to obtain a fibrous product.
[0047] Example 6
[0048] The present invention provides a preparation method of a co-assembled fibrous drug carrier, which comprises: firstly dissolving a Chinese medicine monomer rutin in 10 ml of anhydrous ethanol (3 mg / ml) and dissolving it thoroughly to prepare a solution A. Dissolving a certain amount of tannic acid (3 mg / ml) in pure water to prepare a solution B. Dropping solution A into solution B and stirring to dissolve, the volume ratio of solution A to solution B is 1:4, and ultrasonically removing bubbles, refrigerating at 4°C and standing for 3 days. Centrifuging the above-mentioned mixed solution at 5000 revolutions, washing the solid product with pure water and collecting it, freezing it at -80°C for 48 hours, and freeze-drying to obtain a fibrous product.
[0049] Example 7
[0050] The present invention provides a preparation method of a co-assembled fibrous drug carrier, which comprises: firstly dissolving monomer paclitaxel in 10 ml of anhydrous ethanol (3 mg / ml) and dissolving thoroughly to prepare solution A. Dissolving a certain amount of tannic acid (3 mg / ml) in pure water to prepare solution B. Dropping solution A into solution B and stirring to dissolve, the volume ratio of solution A to solution B is 1:4, and ultrasonically removing bubbles, refrigerating at 4°C and standing for 3 days. Centrifuging the mixed solution at 5000 rpm, washing the solid product with pure water and collecting it, freezing it at -80°C for 48 hours, and freeze-drying to obtain a fibrous product.
[0051] Example 8
[0052] The present invention provides a preparation method of a co-assembled fibrous drug carrier, which comprises: firstly dissolving quercetin, a flavonoid Chinese medicine monomer, in 10 ml of anhydrous ethanol (3 mg / ml), and dissolving it thoroughly to prepare solution A. Dissolving a certain amount of tannic acid (1 mg / ml) in pure water to prepare solution B. Adding solution A dropwise to solution B and stirring to dissolve, the volume ratio of solution A to solution B is 1:5, and ultrasonically removing bubbles, refrigerating at 4°C and standing for 3 days. Centrifuging the above-mentioned mixed solution at 5000 rpm, washing the solid product with pure water and collecting it, freezing it at -80°C for 48 hours, and freeze-drying to obtain a fibrous product.
[0053] Example 9
[0054] The present invention provides a preparation method of a co-assembled fibrous drug carrier, which comprises: firstly dissolving quercetin, a flavonoid Chinese medicine monomer, in 10 ml of anhydrous ethanol (3 mg / ml), and dissolving it thoroughly to prepare solution A. Dissolving a certain amount of tannic acid (1 mg / ml) in pure water to prepare solution B. Adding solution A dropwise to solution B and stirring to dissolve, the volume ratio of solution A to solution B is 1:6, and ultrasonically removing bubbles, refrigerating at 4°C and standing for 3 days. Centrifuging the above-mentioned mixed solution at 5000 rpm, washing the solid product with pure water and collecting it, freezing it at -80°C for 48 hours, and freeze-drying to obtain a fibrous product.
[0055] Example 10
[0056] The present invention provides a preparation method of a co-assembled fibrous drug carrier, which comprises: firstly dissolving quercetin, a flavonoid Chinese medicine monomer, in 10 ml of anhydrous ethanol (3 mg / ml), and dissolving it thoroughly to prepare solution A. Dissolving a certain amount of tannic acid (1 mg / ml) in pure water to prepare solution B. Adding solution A dropwise to solution B and stirring to dissolve, wherein the volume ratio of solution A to solution B is 1:8, and ultrasonically removing bubbles, refrigerating at 4°C and standing for 3 days. Centrifuging the above-mentioned mixed solution at 5000 rpm, washing the solid product with pure water and collecting it, freezing it at -80°C for 48 hours, and freeze-drying to obtain a fibrous product.
[0057] Embodiment 11
[0058] The present invention provides a preparation method of a co-assembled fibrous drug carrier, which comprises: firstly dissolving quercetin, a flavonoid Chinese medicine monomer, in 10 ml of anhydrous ethanol (3 mg / ml), and dissolving it thoroughly to prepare solution A. Dissolving a certain amount of tannic acid (1 mg / ml) in pure water to prepare solution B. Adding solution A dropwise to solution B and stirring to dissolve, the volume ratio of solution A to solution B is 1:10, and ultrasonically removing bubbles, refrigerating at 4°C and standing for 3 days. Centrifuging the mixed solution at 5000 rpm, washing the solid product with pure water and collecting it, freezing it at -80°C for 48 hours, and freeze-drying to obtain a fibrous product.
[0059] The surface images of the fibrous drug carriers provided in Examples 1, 2, 3, 4, and 5 of the present invention are as follows: Figure 1 As shown, the amount of drug precipitation from the carrier provided by Example 1 of the present invention is more obvious, indicating that the amount of carrier precipitation is related to the concentration of tannic acid and quercetin.
[0060] The microscopic images of the carrier drugs provided in Examples 1, 2, 3, 4 and 5 of the present invention are as follows: Figure 2 As shown, field emission electron microscopy observations indicate that the carrier drug provided by our preparation method mainly presents a fibrous structure.
[0061] Example 1 of the present invention, the x-ray diffraction pattern of tannic acid and quercetin, as shown in Figure 3 As shown in FIG. 1 , after placing the material in an x-ray diffractometer, a peak is shown at a rotation angle of 10-80°, representing a crystalline form. Figure 3 As shown, tannic acid has no peak, while quercetin and Example 1 indicate that quercetin and tannic acid are successfully assembled.
[0062] The drug loading rate statistics provided in Examples 1, 2, 3, 4 and 5 of the present invention are as follows: Figure 4 As shown, the results show that the drug loading of quercetin is related to the tannic acid content; however, excessive tannic acid will destroy the assembly of quercetin, resulting in reduced drug loading efficiency. This indicates that excessive tannic acid will weaken the interaction between quercetin and tannic acid, and the concentration shown in Example 1 is the optimal drug loading concentration.
[0063] The encapsulation efficiency statistics provided in Examples 1, 2, 3, 4 and 5 of the present invention are as follows: Figure 5 As shown, the results show that the encapsulation amount of quercetin is related to the tannic acid content, and the concentration shown in Example 1 is the optimal drug encapsulation concentration.
[0064] The cell activity statistics provided in Examples 1, 2, 3, 4 and 5 of the present invention are as follows: Figure 6As shown, within 1 day, the cell activity was detected by CCK-8 kit. Compared with the control group, all samples showed 100% cell viability, which indicates that the embodiments of the present invention have excellent cell compatibility.
[0065] The microscopic image provided by Example 6 of the present invention is as follows: Figure 7 As shown, the tannic acid / rutin co-assembled drug carrier prepared by the same method as in Example 1. Field emission electron microscopy observation revealed that the microstructure of the drug carrier shown in Example 6 was consistent with that in Example 1, showing a fibrous product. This indicates that the scheme described in the present invention has excellent feasibility.
[0066] Zeta potential diagram of the raw materials tannic acid and quercetin of the present invention. Figure 8 As shown in the figure, the potential of tannic acid and quercetin solution was tested by dynamic light scattering, and the results showed that tannic acid was negatively charged and quercetin was positively charged, which indicated that there might be a potential interaction between the two.
[0067] The potential diagrams provided in Examples 1, 2, 3, 4 and 5 of the present invention show that tannic acid exhibits a negative charge and quercetin exhibits a positive charge ( Figure 8 ), and the dynamic light scattering instrument further showed the charge effect of the carrier. Fig. 9 As shown, it was found that Example 1, Example 2, Example 3, Example 4, and Example 5 mainly presented negative charges, which was due to the excessive presence of tannic acid, also indicating the successful co-assembly of tannic acid and quercetin.
[0068] Application Example 1
[0069] Anti-inflammatory treatment of chronic diabetic wounds that are difficult to heal is an important aspect of its treatment. Due to the high sugar environment and immune dysfunction, diabetic wounds often show excessive chronic inflammation. Studies have shown that these hydrogels can reduce the production of inflammatory factors, thereby promoting wound healing. Therefore, we believe that the prepared carrier drug has a stronger anti-inflammatory ability and can be used as an anti-inflammatory drug to promote wound repair. Therefore, we loaded the carrier drug prepared in Example 1 into a gelatin film (a recognized non-toxic matrix, Gel) to prepare a gelatin film loaded with Example 1 (RQ-Gel), and applied it to chronic diabetic wounds that are difficult to heal. The commercial gel (URGO) was used as a positive control, and the repair ability of the prepared material on chronic diabetic wounds that are difficult to heal was evaluated by evaluating its healing ability in vivo for 14 days. The results are as follows. Fig.10 As shown, the wound healing area of the RQ-Gel prepared by us is smaller than that of other groups, and there is a significant variance with the control group and the Gel group. This shows that Example 1 of this project has significant anti-inflammatory ability.
[0070] The inflammatory response between the groups was evaluated by immunofluorescence staining of the tissue. iNOS is a specific indicator of inflammation. We performed fluorescence intensity statistics on the tissue and found that the inflammatory response of the prepared RQ-Gel wound was smaller than that of other groups, and there was a significant variance with the control group and the Gel group. This shows that Example 1 of the present invention has significant anti-inflammatory ability.
Claims
1. A method for preparing a co-assembled fibrous drug carrier, characterized in that: The following steps are involved: S1. First, dissolve quercetin in anhydrous ethanol to prepare solution A; S2, dissolving tannic acid in water to prepare solution B; S3. Add solution B to solution A and stir to dissolve, remove bubbles by ultrasonication, refrigerate and let stand, and obtain a fibrous product through post-treatment as a fibrous drug carrier.
2. The method for preparing the co-assembled fibrous drug carrier according to claim 1, characterized in that: In step S1, the concentration of quercetin in the solution A is 1-6 mg / ml.
3. The method for preparing the co-assembled fibrous drug carrier according to claim 1, characterized in that: In step S2, the concentration of tannic acid in the solution B is 1-6 mg / ml.
4. The method for preparing the co-assembled fibrous drug carrier according to claim 1, characterized in that: In step S3, the volume ratio of solution A to solution B is 1:4-10.
5. The method for preparing the co-assembled fibrous drug carrier according to claim 1, characterized in that: In step S3, the refrigerated storage conditions are: refrigerated storage at 2-6° C. for 2-5 days.
6. The method for preparing the co-assembled fibrous drug carrier according to claim 1, characterized in that: In step S3, the post-processing includes: The mixed solution after refrigeration and standing is centrifuged at 4000-6000 rpm, and the solid product is washed with water and collected, and freeze-dried to obtain a fibrous product.
7. The method for preparing the co-assembled fibrous drug carrier according to claim 6, characterized in that: The freeze-drying conditions are: freeze-drying at -85°C to -75°C for 24-48h.
8. A fibrous drug carrier prepared according to the preparation method according to any one of claims 1 to 7.
9. Use of the fibrous drug carrier according to claim 8 in preparing drugs for treating inflammation.