A pectin-based injectable hydrogel and its preparation method and application
Patent Information
- Application Number
- CN202311073292.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2023-08-23
- Publication Date
- 2026-09-22
- Estimated Expiration
- 2043-08-23
AI Technical Summary
[0005]为了解决上述技术问题，本发明的目的是提供一种果胶基可注射水凝胶及其制备方法和应用，醚化改性过程创新性的克服了高碘酸盐氧化法导致的开环反应，保留了果胶醛分子链的刚性，从而有利于改善果胶基功能材料的力学性能，有效解决了现有技术中高碘酸盐氧化法制备的果胶基可注射水凝胶缓释性能差和力学性能差等问题
[0027]1、本发明醚化改性过程创新性的克服了高碘酸盐氧化法导致的开环反应，保留了果胶醛分子链的刚性，有效解决了现有技术中高碘酸盐氧化法制备的果胶基可注射水凝胶缓释性能差和力学性能差等问题；极大地提高了果胶衍生物的化学反应速率，可与各种含氨基和亚氨基地化合物反应制备水凝胶，有效克服了高碘酸盐氧化果胶基水凝胶在成孔方面存在的问题。
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Abstract
Description
Technical Field
[0001] This invention relates to the field of biomedical materials technology, specifically to a pectin-based injectable hydrogel, its preparation method, and its application. Background Technology
[0002] Cancer is a malignant disease threatening human health. With the aging of the world's population and changes in living environment and daily habits, the number of people dying from cancer is increasing dramatically every year. Cancer treatment has become a crucial task for medical researchers. Chemotherapy, a conventional treatment method, plays a vital and irreplaceable role in cancer treatment. However, while chemotherapy kills cancer cells, it also damages normal cells, producing significant side effects. Many candidate compounds or drugs currently developed or used clinically for cancer treatment have numerous problems, including low bioavailability, instability, significant toxicity and side effects, and lack of targeting, failing to fully meet the clinical needs of cancer treatment. Therefore, finding efficient drug delivery methods is an urgent priority for cancer treatment.
[0003] Pectin is a class of acidic heteropolysaccharides widely found in plant cell walls, linked by D-galacturonic acid via α-1,4-glycosidic bonds. It is characterized by its natural, green, and nutritious properties. Pectin possesses excellent properties such as antibacterial activity, good biocompatibility, hemostasis, detoxification, lipid-lowering effects, and anti-radiation properties, making it widely used in numerous industries including food, health products, cosmetics, and pharmaceuticals. Among these, pectin-based hydrogel materials are currently a research hotspot. Researchers mainly obtain pectin-based hydrogel materials through physical blending or chemical cross-linking. Generally, hydrogel materials formed by physical processes exhibit significant hydrophilic swelling or solubility, limiting their application range. Chemically cross-linked pectin-based hydrogel materials can resist the degradation of hydrogel-based functional materials by water absorption. However, the main functional groups in pectin molecules are hydroxyl or carboxyl groups, resulting in poor reactivity.
[0004] Developing pectin derivatives with high chemical reactivity is a research hotspot. However, periodate oxidation is currently the only method for preparing pectin aldehydes. This method has advantages such as mild preparation conditions, but the ring-opening reaction of D-galacturonic acid leads to a decrease in the mechanical properties of pectin derivatives, thus affecting the application range of pectin-based functional materials. Summary of the Invention
[0005] To address the aforementioned technical problems, the present invention aims to provide a pectin-based injectable hydrogel, its preparation method, and its application. The etherification modification process innovatively overcomes the ring-opening reaction caused by the periodate oxidation method, retaining the rigidity of the pectin aldehyde molecular chain, thereby improving the mechanical properties of pectin-based functional materials. This effectively solves the problems of poor sustained-release performance and poor mechanical properties of pectin-based injectable hydrogels prepared by the periodate oxidation method in the prior art.
[0006] The technical solution of this invention to solve the above-mentioned technical problems is as follows: A method for preparing a pectin-based injectable hydrogel is provided, comprising the following steps:
[0007] (1) Pectin was placed in an alkaline solution and stirred to disperse it, and a pectin dispersion was obtained. Then, cyclopropane-formaldehyde was slowly added to react, and an acidic solution was added to adjust the pH value to 6-8. The product was then centrifuged, washed and freeze-dried to obtain the etherified pectin product.
[0008] (2) Mix the etherified pectin product obtained in step (1) with an amino acid solution, adjust the pH value to 6-8 and stir, react, and let stand at room temperature to obtain a pectin-based injectable hydrogel.
[0009] Furthermore, in step (1), the molecular weight of pectin is 2000-400000 Da.
[0010] Furthermore, in step (1), the alkaline solution is an aqueous solution of sodium hydroxide, an aqueous solution of potassium hydroxide, an aqueous solution of lithium hydroxide, an ethanolic solution of sodium ethoxide, an ethanolic solution of potassium ethoxide, an ethanolic solution of sodium methoxide, or an ethanolic solution of potassium methoxide.
[0011] Furthermore, in step (1), the concentration of the alkaline solution is 1-40 wt%, and the concentration of the pectin dispersion is 0.5-20 wt%.
[0012] Furthermore, in step (1), the mixture is stirred and dispersed for 0.5-4 hours.
[0013] Furthermore, in step (1), the mass-to-volume ratio of pectin to cyclopropane formaldehyde is 0.1-10 g: 0.1-5 mL.
[0014] Furthermore, in step (1), the reaction is carried out at a temperature of 10-60℃ for 0.5-8h.
[0015] Furthermore, in step (1), the acidic solution is an aqueous solution of hydrochloric acid, phosphoric acid, formic acid, acetic acid, acetic acid, or citric acid.
[0016] Furthermore, in step (1), the concentration of the acidic solution is 0.1-5 mol / L.
[0017] Furthermore, in step (1), the sample is washed 3-5 times with a 60-100 wt% aqueous ethanol solution or an aqueous methanol solution.
[0018] Furthermore, in step (1), the mass-to-volume ratio of pectin to aqueous ethanol or aqueous methanol solution is 1 g: 3-10 mL.
[0019] Furthermore, in step (2), the mass ratio of the etherified pectin product to the amino acid solution is 0.5-5:0.5-5.
[0020] Furthermore, in step (2), the concentration of the etherified pectin product is 0.1-10 wt%, and the concentration of the amino acid solution is 0.1-20 wt%.
[0021] Furthermore, in step (2), the amino acid is lysine, arginine, or histidine.
[0022] Further, in step (2), the pH value is adjusted to 6-8 and stirred for 0.5-3h, and the reaction is carried out at 15-90℃ for 10min-5h.
[0023] The above-mentioned methods for preparing etherified pectin products can also be used to prepare aldehyde derivatives of cellulose, starch, sodium alginate, lignin, guar gum, and hemicellulose; the methods for preparing pectin-based injectable hydrogels can also be extended to one or more of albumin, keratin, insulin, glycoproteins, phosphoproteins, lipoproteins, metalloproteins, nucleoproteins, keratin, collagen, zein, catfish antimicrobial peptides, and chitosan.
[0024] The pectin-based injectable hydrogel prepared by the above method is a pectin-based injectable hydrogel.
[0025] Application of the above-mentioned pectin-based injectable hydrogels in drug delivery systems.
[0026] The present invention has the following beneficial effects:
[0027] 1. The etherification modification process of this invention innovatively overcomes the ring-opening reaction caused by the periodate oxidation method, retains the rigidity of the pectin aldehyde molecular chain, and effectively solves the problems of poor sustained-release performance and poor mechanical properties of pectin-based injectable hydrogels prepared by the periodate oxidation method in the prior art; it greatly improves the chemical reaction rate of pectin derivatives, and can react with various amino and imino compounds to prepare hydrogels, effectively overcoming the problems of pore formation in periodate oxidized pectin-based hydrogels.
[0028] 2. This invention innovatively overcomes the ring-opening reaction caused by periodate oxidation during the etherification modification process, preserving the rigidity of the pectin aldehyde molecular chain, thus benefiting the mechanical strength of pectin-based functional materials. The injectable hydrogel is prepared from pectin aldehyde (cyclopropane-formaldehyde modified pectin) using a diamino amino acid as a crosslinking agent. The hydrogel provided by this invention improves upon the existing technical problems of low mechanical properties and limited application range of pectin. The self-healing injectable hydrogel provided by this invention is based on the dynamic equilibrium of Schiff base bonds, and can rapidly self-heal without any external stimulation. It not only has good pH and temperature responsiveness, but also good mechanical properties and good cell compatibility, making it suitable as a carrier for drug delivery systems.
[0029] 3. This invention uses pectin as a raw material and prepares etherified pectin through a ring-opening reaction of cyclopropane-formaldehyde under alkaline conditions. The resulting pectin aldehyde retains the cyclic chemical structure of the main chain, i.e., the rigidity of the main chain, which greatly improves the reactivity of pectin derivatives and expands the application fields of pectin. On the other hand, the aldehyde group has high chemical activity and can react with various amino-containing compounds. Based on the Schiff base reaction, a series of injectable hydrogel-based drug sustained-release systems were prepared. This hydrogel not only has good biocompatibility but also reduces the toxicity of drugs to the body and, as a synergistic antitumor drug delivery carrier, exhibits controllable drug release behavior. The results show that the pectin-based self-healing hydrogel has injectable and self-healing properties and has the potential for application in local antitumor therapy. Attached Figure Description
[0030] Figure 1 The NMR spectrum of the etherified pectin product obtained in Example 1;
[0031] Figure 2 The graph shows the cumulative release rate of the pectin-based injectable hydrogels obtained in Examples 2-4 in simulated small intestinal fluid.
[0032] Figure 3 The graph shows the cumulative release rate of the pectin-based injectable hydrogels obtained in Examples 2-4 in simulated colonic fluid;
[0033] Figure 4 This is a diagram of the cytotoxicity experiment of the pectin-based injectable hydrogel obtained in Example 2;
[0034] Figure 5 This is a diagram of the cytotoxicity experiment of the pectin-based injectable hydrogel obtained in Example 3;
[0035] Figure 6 This is a diagram of the cytotoxicity experiment of the pectin-based injectable hydrogel obtained in Example 4. Detailed Implementation
[0036] The principles and features of this invention are described below. The examples given are for illustrative purposes only and are not intended to limit the scope of the invention. Unless otherwise specified in the examples, conventional conditions or conditions recommended by the manufacturer should be followed. Reagents or instruments whose manufacturers are not specified are all commercially available products.
[0037] Example 1
[0038] An etherified pectin product, the preparation method of which includes the following steps:
[0039] 5g of pectin was placed in 300mL of 10wt% alkaline solution and stirred for 1h to obtain a pectin dispersion. Then, 2mL of cyclopropane formaldehyde was slowly added and reacted at 50℃ for 6h. The pH was adjusted to 6-8 by adding 1mol / L acetic acid, centrifuged, washed 5 times with 30mL of 90wt% ethanol aqueous solution, and freeze-dried to obtain the etherified pectin product.
[0040] Example 2
[0041] A pectin-based injectable hydrogel, the preparation method of which includes the following steps:
[0042] The etherified pectin product (10 wt%) obtained in Example 1 was mixed with lysine solution (20 wt%) at a mass ratio of 1:1, the pH was adjusted to 6-8 and stirred for 2 hours, reacted at 60°C for 3 hours, and allowed to stand at room temperature for 6 hours to obtain pectin-based injectable hydrogel.
[0043] Example 3
[0044] A pectin-based injectable hydrogel, the preparation method of which includes the following steps:
[0045] The etherified pectin product (10 wt%) obtained in Example 1 was mixed with lysine solution (20 wt%) at a mass ratio of 1:2, the pH was adjusted to 6-8 and stirred for 2 hours, reacted at 60°C for 3 hours, and allowed to stand at room temperature for 6 hours to obtain pectin-based injectable hydrogel.
[0046] Example 4
[0047] A pectin-based injectable hydrogel, the preparation method of which includes the following steps:
[0048] The etherified pectin product (10 wt%) obtained in Example 1 was mixed with lysine solution (20 wt%) at a mass ratio of 2:1, the pH was adjusted to 6-8 and stirred for 2 h, reacted at 60 °C for 3 h, and allowed to stand at room temperature for 6 h to obtain pectin-based injectable hydrogel.
[0049] Obtain the NMR spectrum of the etherified pectin product obtained in Example 1, as follows: Figure 1 As shown.
[0050] Depend on Figure 1 It can be seen that the signal at 1.72 ppm is a proton of the methylene group; the signal at 0.73 ppm is a proton of the methyl group; and the signal at 5.67 ppm is a proton of the enol group, further confirming the successful etherification modification. The aldehyde content was calculated to be 0.8095 mmol / g using the following formula.
[0051]
[0052] Wherein: MA has a molecular weight of 116.07; ICPA and I MA The integral areas representing the protons of the methylene group in CPA and the olefin protons in MA are 2 and 2, respectively; m CPA and m MA These correspond to the doses used in quantitative NMR characterization.
[0053] Obtain the cumulative release rate graphs of the pectin-based injectable hydrogels obtained in Examples 2-4 in simulated small intestinal and colonic fluids, as shown below. Figure 2-3 As shown.
[0054] Depend on Figure 2-3 It can be seen that the cumulative release rates of the pectin-based injectable hydrogels obtained in Examples 2-4 after 6 hours of release in simulated small intestinal fluid were 63.65%, 88.33%, and 31.12%, respectively; and the cumulative release rates after 6 hours of release in simulated colonic fluid were 61.16%, 85.37%, and 29.21%, respectively.
[0055] Obtain the cytotoxicity test results of the pectin-based injectable hydrogels obtained in Examples 2-4, such as... Figure 4-6 As shown.
[0056] Depend on Figure 4-6 It can be seen that in the pectin-based injectable hydrogels obtained in Examples 2-4, when the carrier concentration is 3.6 mg / mL, 1.8 mg / mL, 0.9 mg / mL, 0.45 mg / mL and 0.225 mg / mL, the cell proliferation rate of the carrier sample after 72 hours is 80%, indicating that the pectin-based injectable hydrogels obtained in this invention have good biocompatibility and are non-toxic to the body.
[0057] In summary, the pectin-based injectable hydrogel of this invention utilizes the property of shear strain / stress thinning to inject gel blocks into target locations, and the gel blocks rapidly recover to the overall gel phase after injection. This direct injection method eliminates the need to consider gelation time, and its self-healing properties extend its lifespan. Therefore, the self-healing pectin-based injectable hydrogel avoids the potential risk of drug diffusion, thereby effectively utilizing drugs and reducing toxicity to normal tissues.
[0058] The above description is only a preferred embodiment of the present invention and is 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 within the protection scope of the present invention.
Claims
1. A method for preparing a pectin-based injectable hydrogel, characterized in that, Includes the following steps: (1) Pectin was placed in an alkaline solution and stirred to disperse it, and a pectin dispersion was obtained. Then cyclopropane formaldehyde was added, and cyclopropane formaldehyde underwent a ring-opening reaction and was grafted onto the pectin molecular chain, retaining the cyclic structure of the pectin molecular chain. An acidic solution was added to adjust the pH value to 6-8, and then the product was centrifuged, washed and freeze-dried to obtain the etherified pectin product. (2) The etherified pectin product obtained in step (1) is mixed with an amino acid solution, wherein the amino acid is lysine, arginine or histidine, and a Schiff base crosslinking reaction is carried out with the etherified pectin product to form a dynamic covalent network; the pH value is adjusted to 6-8 and stirred, the reaction is carried out, and the mixture is allowed to stand at room temperature to obtain a pectin-based injectable hydrogel.
2. The method for preparing pectin-based injectable hydrogel as described in claim 1, characterized in that, In step (1), the alkaline solution is an aqueous solution of sodium hydroxide, an aqueous solution of potassium hydroxide, an aqueous solution of lithium hydroxide, an aqueous solution of sodium ethoxide, an aqueous solution of potassium ethoxide, an aqueous solution of sodium methoxide, or an aqueous solution of potassium methoxide.
3. The method for preparing pectin-based injectable hydrogel as described in claim 1, characterized in that, In step (1), the mass-to-volume ratio of pectin to cyclopropane formaldehyde is 0.1-10g:0.1-5mL.
4. The method for preparing pectin-based injectable hydrogel as described in claim 1, characterized in that, In step (1), the acidic solution is an aqueous solution of hydrochloric acid, phosphoric acid, formic acid, acetic acid, acetic acid, or citric acid.
5. The method for preparing pectin-based injectable hydrogel as described in claim 1, characterized in that, In step (2), the mass ratio of the etherified pectin product to the amino acid solution is 0.5-5:0.5-5.
6. The method for preparing pectin-based injectable hydrogel as described in claim 1, characterized in that, In step (2), the concentration of the etherified pectin product is 0.1-10 wt%, and the concentration of the amino acid solution is 0.1-20 wt%.
7. The method for preparing pectin-based injectable hydrogel as described in claim 1, characterized in that, In step (2), adjust the pH value to 6-8 and stir for 0.5-3h, and react at 15-90℃ for 10min-5h.
8. The pectin-based injectable hydrogel prepared by the method of any one of claims 1-7.
9. The use of the pectin-based injectable hydrogel according to claim 8 in the preparation of an antitumor drug delivery carrier.