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CN-122011237-A - Melatonin-loaded double-crosslinked hydrogel and application thereof

CN122011237ACN 122011237 ACN122011237 ACN 122011237ACN-122011237-A

Abstract

The invention relates to a melatonin-loaded double-crosslinked hydrogel and application thereof. The invention takes polylysine grafted with gallic acid (PL-GA) as an intelligent carrier, takes hyaluronic acid grafted with phenylboronic acid and glycidyl methacrylate (HA-PBA-GMA) as a responsive network skeleton and a stable skeleton precursor, loads melatonin on the PL-GA to form a drug-loaded functional unit (MT@ PL-GA), mixes MT@ PL-GA with HA-PBA-GMA solution, and prepares the double-crosslinked hydrogel with environmental responsiveness and structural stability through 'dynamic boric acid ester bond crosslinking' and 'photoinitiated free radical polymerization'. The double-crosslinked hydrogel has high melatonin loading rate, excellent stability and self-healing performance, and mechanical properties matched with the wound surface mechanical environment, and can release medicines aiming at the responsiveness of the wound microenvironment, so that the double-crosslinked hydrogel is very suitable for the treatment of local wounds.

Inventors

  • XIE YOUFU
  • HU PING
  • WEI WENLONG
  • NING XUYANG
  • LU HONG

Assignees

  • 广州市红十字会医院

Dates

Publication Date
20260512
Application Date
20260318

Claims (10)

  1. 1. A modified hyaluronic acid is characterized in that the modified hyaluronic acid is obtained by reacting hyaluronic acid grafted with phenylboronic acid groups with glycidyl methacrylate, The grafting rate of phenylboronic acid groups in the modified hyaluronic acid is 35% -55%; the grafting rate of the glycidyl methacrylate group in the modified hyaluronic acid is 15-35%.
  2. 2. The modified hyaluronic acid of claim 1, wherein the weight average molecular weight of the hyaluronic acid is between 400kDa and 800kDa; And/or the grafting ratio of phenylboronic acid groups in the modified hyaluronic acid is 40% -50%, more preferably 43% -50%; And/or the grafting ratio of the glycidyl methacrylate groups in the modified hyaluronic acid is 20% -32%, more preferably 23% -31%.
  3. 3. A method for producing the modified hyaluronic acid according to claim 1 or 2, characterized by comprising the steps of: Carrying out amidation reaction on hyaluronic acid and 3-aminophenylboronic acid to obtain hyaluronic acid grafted with phenylboronic acid groups; Performing epoxy ring-opening reaction on the hyaluronic acid grafted with the phenylboronic acid group and glycidyl methacrylate to obtain the modified hyaluronic acid; preferably, the molar ratio of saccharide units in the hyaluronic acid to 3-aminophenylboronic acid is from 1:0.7 to 0.85, preferably 1:0.8; Preferably, the molar ratio of saccharide units in the hyaluronic acid to glycidyl methacrylate is 1:0.5-0.7, preferably 1:0.6.
  4. 4. A melatonin-loaded double crosslinked hydrogel precursor solution, comprising solution a and solution B; the solution a consists of the modified hyaluronic acid of claim 1 or 2, a photoinitiator and a solvent; The solution B consists of a melatonin-carrying polymer and a solvent; the melatonin-loaded polymer is prepared by loading melatonin by modified polylysine; The modified polylysine is obtained by amidation reaction of epsilon-polylysine and gallic acid; preferably, the solution A and the solution B are uniformly mixed according to the volume ratio of 1:0.9-1.1 before use.
  5. 5. The melatonin-loaded double crosslinked hydrogel precursor solution of claim 4, wherein the grafting ratio of gallic acid in the modified polylysine is 2.8% -23%, preferably 12% -18%, more preferably 14% -17%; and/or the epsilon-polylysine and gallic acid are reacted in a molar ratio of amino groups in epsilon-polylysine to gallic acid of 1:0.8-2, preferably 1:1.2-1.8, more preferably 1:1.4-1.6; and/or the epsilon-polylysine has a weight average molecular weight of 3000-5000; and/or the melatonin loading of the melatonin-loaded polymer is 4% -15%, preferably 9% -12%; And/or the solvent in the solution B is water or a buffer, preferably PBS buffer with pH of 7.4; and/or the concentration of the solution B is 0.25g/mL-0.35g/mL, preferably 0.28g/mL-0.32g/mL, based on the mass of the modified polylysine.
  6. 6. The melatonin-loaded double crosslinked hydrogel precursor solution of claim 4 or 5, wherein the photoinitiator is phenyl (2, 4, 6-trimethylbenzoyl) lithium phosphate; and/or the solvent in the solution A is water or a buffer, preferably PBS buffer with pH of 7.4; And/or the concentration of the modified hyaluronic acid in the solution A is 90 mg/mL-110 mg/mL; And/or the concentration of the photoinitiator in the solution A is 0.5 mg/mL-2 mg/mL.
  7. 7. A method of preparing a melatonin-loaded double crosslinked hydrogel precursor solution of any one of claims 4-6, comprising the steps of: preparing a solution A, namely dissolving the modified hyaluronic acid in a solvent, adding the photoinitiator, and stirring and dissolving in a dark place to obtain the modified hyaluronic acid; Preparing a solution B, namely dissolving melatonin in ethanol to obtain a melatonin ethanol solution, dissolving the modified polylysine in water or buffer solution to obtain a polymer solution, dripping the melatonin ethanol solution into the polymer solution, continuously stirring after dripping, standing, and removing ethanol to obtain the melatonin; Preferably, the mass ratio of melatonin to modified polylysine is 1:5-15, preferably 1:7-10, more preferably 1:8-9; Preferably, the concentration of melatonin in the melatonin ethanol solution is 15mg/mL-25 mg/mL; preferably, the buffer is a PBS buffer at pH 7.4; Preferably, the concentration of the modified polylysine in the polymer solution is 0.25g/mL-0.35g/mL, preferably 0.28g/mL-0.32g/mL; Preferably, the melatonin-ethanol solution is added dropwise to the polymer solution at a rate of 0.4mL/min-0.6 mL/min; preferably, the melatonin-ethanol solution is added dropwise to the polymer solution at a stirring speed of 120 rpm-180 rpm; Preferably, stirring is continued for 3-6 hours after the dripping is completed; preferably, the mixture is left to stand for 10 hours to 16 hours.
  8. 8. The preparation method of the melatonin-loaded double-crosslinked hydrogel is characterized by comprising the following steps of uniformly mixing a solution A and a solution B in the melatonin-loaded double-crosslinked hydrogel precursor solution according to any one of claims 4-6, standing, and irradiating with ultraviolet light to obtain the melatonin-loaded double-crosslinked hydrogel; preferably, the solution A and the solution B are uniformly mixed according to the volume ratio of 1:0.9-1.1; Preferably, the time of irradiation with ultraviolet light is 0 to 60 seconds, preferably 28 to 32 seconds; Preferably, the standing is at 35-40 ℃ for 3-5 minutes; Preferably, the wavelength of the ultraviolet light is 365 nm, and the light intensity is 4mW/cm2-6 mW/cm2.
  9. 9. The melatonin-loaded double crosslinked hydrogel prepared by the method of claim 8.
  10. 10. Use of the melatonin-loaded double crosslinked hydrogel precursor solution of any of claims 4-6 and/or the melatonin-loaded double crosslinked hydrogel of claim 9 in the preparation of a medicament, The medicament is an anti-inflammatory medicament, an antioxidant medicament, a medicament for promoting wound healing and/or inhibiting scarring.

Description

Melatonin-loaded double-crosslinked hydrogel and application thereof Technical Field The invention belongs to the technical field of pharmaceutical preparations, relates to hydrogel, and in particular relates to melatonin-loaded double-crosslinked hydrogel and application thereof in wound healing and scar inhibition. Background Melatonin (N-acetyl-5-methoxytryptamine) acts as a pleiotropic indoleamine hormone secreted by the pineal gland, and its physiological function is far from regulating circadian rhythms. In recent years, melatonin and metabolites thereof are powerful endogenous antioxidants and free radical scavengers, can effectively neutralize Reactive Oxygen Species (ROS) and Reactive Nitrogen Species (RNS), and strengthen the endogenous antioxidant defense system of cells by activating pathways such as nuclear factor E2 related factor 2 (Nrf 2). In the complex pathophysiological processes of wound healing, excessive oxidative stress is a key factor leading to persistent inflammation, abnormal cell proliferation and deregulation of collagen deposition, ultimately contributing to the formation of pathological scars (such as hypertrophic scars and keloids). Therefore, the melatonin is locally applied to directly act on the wound microenvironment, and can accurately regulate and control the healing process, namely, inhibiting excessive inflammation and oxidative damage in the inflammatory phase, promoting angiogenesis and re-epithelialization in the proliferation phase and regulating the balance of collagen synthesis and degradation in the remodeling phase, thereby inhibiting scar while accelerating healing. However, melatonin has a problem in practical use, first, chemical instability. The indole ring structure is sensitive to light, heat and oxidation environment, and is easy to inactivate when being directly smeared. Second, bioavailability is low. Simple creams or solutions are difficult to effectively reside on moist wounds and have limited permeability. Third, space-time controllability is lacking. The requirements of melatonin in different stages of wound healing are dynamically changed, and conventional dosage forms cannot realize accurate release according to the requirements, so that insufficient curative effect or resource waste can be caused. These problems severely limit their effectiveness in topical wound treatment. Therefore, there is a need to develop a new formulation of melatonin which is suitable for use in the treatment of topical wounds, enhancing its practical effect on wound healing and inhibiting scarring. Disclosure of Invention Based on the above, the invention provides the melatonin-loaded double-crosslinked hydrogel precursor solution and the double-crosslinked hydrogel thereof, and the double-crosslinked hydrogel has excellent stability and self-healing performance, and simultaneously has the mechanical properties matched with the mechanical environment of a wound surface, and the drug release is controllable, so that the melatonin-loaded double-crosslinked hydrogel is very suitable for the treatment of local wounds. The specific technical scheme comprises the following steps of. In a first aspect, the present invention provides a modified hyaluronic acid obtained by reacting hyaluronic acid grafted with phenylboronic acid groups with glycidyl methacrylate, The grafting rate of phenylboronic acid groups in the modified hyaluronic acid is 35% -55%; the grafting rate of the glycidyl methacrylate group in the modified hyaluronic acid is 15-35%. In a second aspect, the present invention provides a method for preparing the modified hyaluronic acid, comprising the steps of: Carrying out amidation reaction on hyaluronic acid and 3-aminophenylboronic acid to obtain hyaluronic acid grafted with phenylboronic acid groups; And (3) carrying out epoxy ring-opening reaction on the hyaluronic acid grafted with the phenylboronic acid group and glycidyl methacrylate to obtain the modified hyaluronic acid. In a third aspect, the present invention provides a melatonin-loaded double crosslinked hydrogel precursor solution comprising solution a and solution B; The solution A consists of the modified hyaluronic acid, a photoinitiator and a solvent; The solution B consists of a melatonin-carrying polymer and a solvent; the melatonin-loaded polymer is prepared by loading melatonin by modified polylysine; The modified polylysine is obtained by amidation reaction of epsilon-polylysine and gallic acid. In a fourth aspect, the invention provides a method for preparing the melatonin-loaded double-crosslinked hydrogel precursor solution, comprising the following steps: preparing a solution A, namely dissolving the modified hyaluronic acid in a solvent, adding the photoinitiator, and stirring and dissolving in a dark place to obtain the modified hyaluronic acid; the preparation method comprises the steps of preparing a solution B, dissolving melatonin in ethanol to obtain a melatonin ethanol solution,