Self-assembly curcumin delivery system based on trityl nonaethylene glycol / ovalbumin and preparation method

Abstract

The invention belongs to the technical field of medical, dental or dressing preparations, and particularly discloses a self-assembly curcumin delivery system based on trityl-nonaethylene glycol / ovalbumin, which is obtained by self-assembly of trityl-nonaethylene glycol, ovalbumin and curcumin. The invention also discloses a preparation method of the curcumin delivery system. The drug delivery system prepared by the method obviously improves the encapsulation dispersity and cell absorption efficiency of the drug, and can be widely applied to the fields of food, daily chemicals and medicines.

Description

Technical Field

[0001] This invention belongs to the field of medical, dental or cosmetic formulation technology, specifically relating to a self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin and its preparation method. Background Technology

[0002] Curcumin, an active ingredient found in turmeric, has garnered significant attention in recent years due to its antioxidant, anti-inflammatory, and anti-aging bioactivities. However, its poor water solubility, susceptibility to photodegradation, chemical instability, and low bioavailability limit its application in many functional foods, supplements, and pharmaceuticals. Therefore, there is an urgent need to explore methods such as nanodelivery systems, nanoemulsion encapsulation, chemical modification, and physical dispersion to improve the solubility, stability, and bioavailability of curcumin.

[0003] Patent document CN 117899052 A discloses a method for preparing curcumin nanoparticles as curcumin delivery carriers. The invention utilizes biopolymers such as zein, sodium caseinate, and sodium alginate, employing an antisolvent precipitation technique to prepare nanoparticles that can serve as curcumin delivery carriers. The curcumin nanoparticles prepared by this method primarily address technical issues such as improving the water solubility, antioxidant activity, and photochemical stability of curcumin. However, it makes no mention of how to promote cellular uptake efficiency. Furthermore, the process employed requires the self-assembly of two layers (casein and sodium alginate), which is complex and has low assembly efficiency.

[0004] Ovalbumin (OVA) is abundant in egg white and, as one of the richest protein sources, has attracted widespread attention in the development of delivery systems. Due to its excellent properties such as gelation, easy digestibility, self-assembly, and amphiphilicity, a wide variety of emulsion and gel delivery systems have been constructed based on OVA for the delivery of active substances such as curcumin. However, OVA typically requires heat treatment for denaturation to promote binding with curcumin. Denatured OVA exposes hydrophobic groups, making it prone to flocculation and precipitation, generally requiring further stabilization. OVA particles are spherical, making it difficult for general emulsifiers to bind to the protein surface and form well-dispersed small-scale nanoparticles.

[0005] Triphenylmethyl groups possess advantages such as high selectivity and ease of introduction, and are commonly used in traditional organic chemical and pharmaceutical synthesis as group protection for amines, alcohols, and thiols. However, their self-assembly properties have not yet been utilized. Due to the unique trefoil-like structure formed by its three benzene rings, triphenylmethyl groups can effectively adhere to the surface of globular proteins, making them a promising small-molecule protein particle stabilization structural unit with broad application prospects. Polyethylene glycol (PEG) molecules contain ether bonds and hydroxyl groups, exhibiting good hydrophilicity and biocompatibility. It is widely used for hydrophilic modification of emulsifiers, pharmaceuticals, and materials. PEG is also a commonly used cell fusion inducer, promoting cell fusion by altering cell membrane structure; however, molecular weight typically needs to be 2000 or higher. Summary of the Invention

[0006] To address the problems existing in the prior art, this invention provides a method for preparing a self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin. In this method, triphenylmethyl and nonadecanol are combined via ether bonds to form an amphiphilic molecule. The triphenylmethyl trefoil plate structure effectively binds to the hydrophobic surface of globular ovalbumin through hydrophobic and aromatic ring interactions, significantly improving the encapsulation and dispersibility of the delivery system. The polyethylene glycol structure of nonadecanol (molecular weight ~400, far below 2000) effectively binds to cells, significantly promoting the uptake of nanoparticles.

[0007] A self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin is obtained by self-assembly of triphenylmethylnonadecanol with ovalbumin and curcumin; the structure of the triphenylmethylnonadecanol is as follows:

[0008] .

[0009] Preferably, the delivery system is a self-assembled nanoparticle formed by triphenylmethylnonadecanol, ovalbumin, and curcumin.

[0010] Preferably, the delivery system is obtained by the following method: mixing triphenylmethylnonethylene glycol aqueous solution, ovalbumin aqueous solution and curcumin aqueous solution, heating treatment, cooling, to obtain the self-assembled nanoparticles.

[0011] Preferably, the mass ratio of triphenylmethylnonadecanol, ovalbumin and curcumin is (20~40):100:(20~40).

[0012] Preferably, the triphenylmethylnonadecanol is obtained by reacting triphenylchloromethane and nonadecanol as reaction substrates.

[0013] A method for preparing triphenylmethylnonadecanol includes the following steps:

[0014] Triphenylchloromethane was reacted with nonaglycol molecules as a modifying molecule, and the reaction was completed to obtain a triphenylmethyl derivative.

[0015] Preferably, a base is added during the preparation of triphenylmethylnonadecanol. The base used in the reaction is one of potassium carbonate, DIPEA, triethylamine, imidazole, 2,6-dimethylpyridine, 2,6-pyridine, or potassium hydroxide. To improve the conversion rate and reduce byproducts, triethylamine is further preferred.

[0016] Preferably, the solvent used in the preparation of triphenylmethylnonadecanol is one of DMF, anhydrous acetonitrile, tetrahydrofuran, tert-butanol, dichloromethane, or ethyl acetate. To improve the conversion rate and reduce byproducts, dichloromethane is further preferred.

[0017] A method for preparing a self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin includes: mixing an aqueous solution of triphenylmethylnonadecanol, an aqueous solution of ovalbumin, and a curcumin suspension, heating the mixture, and finally obtaining the self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin.

[0018] Furthermore, using water as a solvent, the ovalbumin solution, curcumin suspension, and triphenylmethylnonadecanol solution were ultrasonically mixed, heated, and then gradually cooled to room temperature, simultaneously completing denaturation and embedding. This yielded the self-assembled nanoparticles.

[0019] Preferably, the average particle size of the self-assembled nanoparticles is 30-100 nanometers. More preferably, the average particle size of the self-assembled nanoparticles is 30-80 nanometers; even more preferably, the average particle size of the self-assembled nanoparticles is 30-70 nanometers.

[0020] Preferably, the heat treatment temperature is 70~100℃. More preferably, the heat treatment temperature is 70~90℃. Even more preferably, the heat treatment temperature is 80~100℃. As the most preferred embodiment, the heat treatment temperature is 90℃.

[0021] Preferably, the heat treatment time is 10 to 30 minutes.

[0022] Preferably, after the heat treatment is completed, the temperature is lowered to 20~30℃ and the reaction continues for 10~25 min to obtain self-assembled nanoparticles.

[0023] Preferably, the mass ratio of triphenylmethylnonaglycol, ovalbumin, and curcumin is (20~40):100:(20~40). Preferably, ovalbumin, triphenylmethylnonaglycol, and curcumin are self-assembled in a mass ratio of 4:1:1 to obtain drug-loaded nanoparticles.

[0024] Preferably, the concentration of the triphenylmethylnonethylene glycol aqueous solution is 0.15~1 mg / mL; and the concentration of the ovalbumin solution is 0.5~2 mg / mL.

[0025] Preferably, the concentration of the curcumin aqueous suspension is 0.15~1 mg / mL. More preferably, the concentration of the curcumin aqueous suspension is 0.15~0.5 mg / mL; even more preferably, the concentration of the curcumin aqueous suspension is 0.15~0.35 mg / mL; even more preferably, the concentration of the curcumin aqueous suspension is 0.2~0.3 mg / mL; specifically, the concentration of the curcumin aqueous suspension is 0.25 mg / mL.

[0026] Preferably, the concentration of the triphenylmethylnonadecanol aqueous solution is 0.15~0.5 mg / mL; more preferably, the concentration of the triphenylmethylnonadecanol aqueous solution is 0.15~0.35 mg / mL; specifically, the concentration of the triphenylmethylnonadecanol aqueous solution is 0.25 mg / mL.

[0027] Preferably, the concentration of the ovalbumin solution is 0.5~2 mg / mL; more preferably, the concentration of the ovalbumin solution is 1~2 mg / mL; specifically, the concentration of the ovalbumin solution is 1 mg / mL.

[0028] This invention uses triphenylmethyl as a modifying group and nonaethylene glycol as a reaction substrate to synthesize a molecule, triphenylmethylnonaethylene glycol, which exhibits good amphiphilicity and biocompatibility. The synthesized molecule is then fully self-assembled with raw materials such as ovalbumin and curcumin under heating conditions to form drug-loaded nanoparticles. The self-assembled nanomedicine delivery system based on triphenylmethylnonaethylene glycol prepared by this invention has an efficient and easy preparation method, significantly improving drug encapsulation dispersion and absorption efficiency, and can be widely used in the food, daily chemical, and pharmaceutical fields.

[0029] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0030] (1) This invention innovatively utilizes the globular protein binding properties of triphenylmethylnonethylene glycol for the preparation of self-assembled nanoparticles, and prepares uniform small molecule drug-loaded self-assembled nanoparticles. The preparation method is simple and easy to implement, and greatly improves the drug encapsulation and dispersion.

[0031] (2) This invention innovatively utilizes the cell binding properties of triphenylmethylnonethylene glycol, and the drug-loaded self-assembled nanoparticles significantly improve cell absorption efficiency, solving the technical problem of low absorption efficiency in traditional methods. Attached Figure Description

[0032] Figure 1Scanning electron microscope (SEM) images of curcumin nanoparticles based on triphenylmethylnonadecanol (EG-OVA-Cur) and curcumin nanoparticles based on ovalbumin (OVA-Cur).

[0033] Figure 2 The graph shows the cell uptake results of EG-OVA-Cur, OVA-Cur, and the control group (CK) obtained by flow cytometry.

[0034] Figure 3 The image shows the H-NMR spectrum of triphenylmethylnonadecanol.

[0035] Figure 4 This is the mass spectrum of triphenylmethylnonadecanol. Detailed Implementation

[0036] To further illustrate the present invention, the invention will be described in more detail below with reference to the accompanying drawings and embodiments, but the implementation of the present invention is not limited thereto.

[0037] Example 1: Preparation of curcumin nanoparticles based on triphenylmethylnonadecanol

[0038] 2.0 eq (2 mmol, 812.73 μL) of nonaethylene glycol was dissolved in 3 mL of dichloromethane, followed by 1.2 eq (166.34 μL) of triethylamine and 1.0 eq (0.2788 μL) of triphenylchloromethane. The mixture was reacted at room temperature for 3 h, and the product, triphenylmethylnonaethylene glycol, was obtained by column chromatography. Using water as a solvent, solutions of 1 mg / mL ovalbumin, 0.25 mg / mL triphenylmethylnonaethylene glycol, and 0.25 mg / mL curcumin were prepared. After ultrasonic mixing, the mixture was heat-treated at 90 °C for 15 min at a 1:1:1 volume ratio, followed by gradual cooling to 25 °C and a further 15 min reaction. This process simultaneously completed denaturation and assembly / embedding. The resulting self-assembled particles were curcumin nanoparticles based on triphenylmethylnonaethylene glycol, denoted as EG-OVA-Cur.

[0039] Among them, the product triphenylmethylnonaglycol 1 1H NMR data (400 MHz, DMSO-d6): δ 7.44–7.16 (m, 15 H), 4.57 (t, J = 5.4 Hz, 1 H), 3.62–3.37 (m, 34 H), 3.06 (t, J = 4.8 Hz, 2 H); ESI mass spectrometry: [M+Na] + 679.52 m / z.

[0040] Ratio optimization: Different concentrations of triphenylmethylnonaethylene glycol (0.03 mg / mL, 0.06 mg / mL, 0.12 mg / mL, 0.25 mg / mL, 0.5 mg / mL, 1 mg / mL), 1 mg / mL ovalbumin solution, and 0.25 mg / mL curcumin suspension were ultrasonically mixed and then heat-treated at 90℃ for 15 min at a 1:1:1 volume ratio. The temperature was then gradually lowered to 25℃ and the reaction continued for another 15 min, simultaneously completing denaturation and assembly / embedding. The particle size of the nanoparticles was measured using a nanoparticle size analyzer, and the particle sizes were 78 nm, 79 nm, 72 nm, 63 nm, 71 nm, and 179 nm. The 0.25 mg / mL triphenylmethylnonaethylene glycol concentration resulted in the smallest particle size and was selected as the optimal ratio.

[0041] Comparative Example 1: Preparation of curcumin nanoparticles based on ovalbumin

[0042] Using water as a solvent, 1 mg / mL ovalbumin solution and 0.25 mg / mL curcumin suspension were prepared. After ultrasonic mixing, the mixture was heat-treated at 90℃ for 15 min at a 1:1 volume ratio, and then gradually cooled to 25℃ for another 15 min to complete denaturation and assembly embedding. The resulting self-assembled particles were ovalbumin-based curcumin nanoparticles, denoted as OVA-Cur.

[0043] Performance Test 1: Evaluation of Dispersion of Triphenylmethyl-Modified Self-Assembled Nanoparticles

[0044] The particle size of the two self-assembled nanoparticles prepared in Example 1 and Comparative Example 1 was measured using a nanoparticle size analyzer. The particle size of EG-OVA-Cur was 63 nm; the particle size of OVA-Cur was 83 nm. The morphology and structure of the two self-assembled nanoparticles prepared in Example 1 and Comparative Example 1 were characterized using a scanning electron microscope. The test results are as follows. Figure 1 As shown, compared with ovalbumin self-assembled curcumin nanoparticles, the self-assembled nanoparticles modified with triphenylmethylnonadecanol have a smoother and more regular surface, clearer and sharper edge contours, and exhibit highly consistent monodispersity and geometric symmetry. They also have smaller and more uniform particle sizes, forming nanoparticles with a dense structure and good uniformity.

[0045] Performance Test 2: Evaluation of Cell Absorption Efficiency of Triphenylmethyl-Modified Self-Assembled Nanoparticles

[0046] Take 30 μL of Saccharomyces cerevisiae culture into a centrifuge tube, add 150 μL of self-assembled nanoparticle suspension (prepared in Example 1), and incubate in a shaker at 200 rpm and 30°C. After incubation for 4 h, stop the reaction, centrifuge at 3000 rpm and 4°C for 3 min to remove the culture medium, then add 50 μL of PBS, centrifuge under the same conditions, and resuspend in PBS.

[0047] After each group of samples was incubated using the above procedures, the fluorescence intensity of the samples was collected using a flow cytometer. Each sample (the negative control group was the cell group without "triphenylmethyl-modified self-assembled nanoparticles", denoted as the CK (Control) group) was diluted with PBS to an appropriate factor before collection. The emission channel of the flow cytometer was set to KO525-A, and each sample was collected 3 times.

[0048] The test results showed that the cell uptake rate (i.e., fluorescence intensity) of ovalbumin-assembled curcumin nanoparticles was 22%, while that of triphenylmethylnonadecanol-assembled curcumin nanoparticles was 32%, indicating a significant increase in cell uptake rate. Figure 2 .

Claims

1. A self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin, characterized in that, It is obtained by self-assembly of triphenylmethylnonaglycol with ovalbumin and curcumin; the structure of the triphenylmethylnonaglycol is as follows: 。 2. The self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin according to claim 1, characterized in that, The delivery system consists of self-assembled nanoparticles formed by triphenylmethylnonadecanol, ovalbumin, and curcumin.

3. The self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin according to claim 2, characterized in that, The mass ratio of triphenylmethylnonadecanol, ovalbumin and curcumin is (20~40):100:(20~40).

4. The self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin according to claim 2, characterized in that, The average particle size of the self-assembled nanoparticles is 30-100 nanometers.

5. A method for preparing a self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin as described in any one of claims 1 to 4, comprising: The triphenylmethylnonadecanol aqueous solution, ovalbumin aqueous solution, and curcumin aqueous suspension were mixed evenly, heated, and then cooled to obtain the self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin.

6. The method for preparing the self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin according to claim 5, characterized in that, The concentration of the triphenylmethylnonadecanol aqueous solution is 0.15~1 mg / mL; the concentration of the ovalbumin aqueous solution is 0.5~2 mg / mL; and the concentration of the curcumin aqueous suspension is 0.15~1 mg / mL.

7. The method for preparing the self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin according to claim 5, characterized in that, The heat treatment temperature is 70~100℃, and the heat treatment time is 10~30 minutes.

8. The method for preparing the self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin according to claim 7, characterized in that, After the heat treatment is completed, the temperature is lowered to 20~30℃ and the reaction continues for 10~25 min to obtain self-assembled nanoparticles.

9. The method for preparing the self-assembled curcumin delivery system based on triphenylmethylnonadecanol / ovalbumin according to claim 5, characterized in that, The triphenylmethylnonadecanol was obtained by reacting triphenylchloromethane and nonadecanol as reaction substrates.

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