Preparation method of resveratrol nano-preparation

Abstract

The present application relates to a preparation method of a nano preparation of resveratrol, and belongs to the technical field of traditional Chinese medicines. In view of the problems of low oral bioavailability and lack of colon targeting of resveratrol, the present application provides a nano preparation composed of resveratrol, zein and xylan. The preparation dissolves resveratrol and zein in an organic phase, drops into the xylan water phase, and removes the organic solvent to obtain the nano preparation. The present application utilizes the hydrophobic effect of zein to improve the drug encapsulation rate, utilizes the colon degradation characteristics and anti-inflammatory activity of xylan to realize synergistic effect, and realizes the precise release of resveratrol at the inflammatory site in the intestinal tract.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine technology, specifically relating to a method for preparing resveratrol nanoparticles. Background Technology

[0002] Inflammatory colitis (ICP) is a chronic, nonspecific inflammatory bowel disease that primarily affects the rectal and colonic mucosa and submucosa. Clinical manifestations include abdominal pain, diarrhea, and bloody, mucous stools. The condition is often chronic and difficult to cure, and carries a risk of malignant transformation. In recent years, with changes in lifestyle and improved diagnostic capabilities, the incidence of ICP in my country has been increasing annually, becoming a research hotspot in the field of digestive system diseases. Currently, clinical treatment for ICP mainly relies on aminosalicylic acid preparations, glucocorticoids, immunosuppressants, and biologics. However, these drugs generally suffer from significant side effects with long-term use, high cost, poor response in some patients, or drug resistance. Therefore, developing novel, highly effective, low-toxicity, and inexpensive treatment strategies is of significant clinical importance.

[0003] Resveratrol is a naturally occurring polyphenol compound widely found in plants such as Polygonum cuspidatum, grapes, and peanuts, possessing various biological activities including anti-inflammatory, antioxidant, and immunomodulatory effects. Recent studies have shown that resveratrol and its derivatives can generally increase the diversity of the gut microbiota and reduce bacterial translocation. It can promote the growth of beneficial bacteria, such as Clostridium perfringens and Bifidobacterium. In addition, resveratrol and its derivatives can regulate various signaling pathways (such as NF-κB) to reduce the infiltration of pro-inflammatory factors and neutrophils (Resveratrol and its derivates improve inflammatory bowel disease by targeting gut microbiota and inflammatory signaling pathways[J]. Food Science and Human Wellness, 2022, 11(1): 22-31).

[0004] However, the clinical application of resveratrol is limited by its poor water solubility, extremely low bioavailability after oral administration, and insufficient blood drug concentration. Furthermore, resveratrol is easily degraded in the acidic environment of the stomach and is difficult to release in the intestines, resulting in most of the drug failing to reach the lesion site in the colon to exert its effect. Achieving efficient intestinal-targeted delivery of resveratrol is a key issue that urgently needs to be addressed in its clinical translation.

[0005] In recent years, nanodelivery systems based on natural biomolecules have provided new insights into improving the oral bioavailability of poorly soluble drugs. Zein, a hydrophobic storage protein extracted from corn, possesses good biocompatibility and degradability. However, single zein nanoparticles exhibit poor stability in the gastrointestinal environment, easily aggregating and precipitating. Xylan, a hemicellulose polysaccharide derived from plant cell walls, possesses multiple bioactivities, including anti-inflammatory and prebiotic properties, and can be specifically degraded by colonic microorganisms, exhibiting natural colon-targeting characteristics.

[0006] Currently, there are no reports on the use of zein and xylan to construct composite nanocarriers for colon-targeted delivery of resveratrol. This invention addresses the shortcomings of existing technologies by, for the first time, constructing a nanocarrier using a combination of zein and xylan to encapsulate resveratrol. The hydrophobic properties of zein enhance drug encapsulation efficiency, while the colonic degradation properties and anti-inflammatory activity of xylan achieve synergistic effects. Simultaneously, the synergistic effect of both resists gastric acid degradation, enabling precise release of resveratrol at sites of intestinal inflammation. This provides a highly efficient and safe new strategy for the treatment of ulcerative colitis. Summary of the Invention

[0007] In view of the shortcomings of the prior art, the present invention provides a method for preparing a nano-formulation of resveratrol. This nano-formulation can simultaneously utilize zein and xylan as carriers, solving the problem of oral delivery of resveratrol, and can be used to treat inflammatory colitis.

[0008] This invention specifically provides a method for preparing resveratrol nanoparticles, wherein the preparation is made from resveratrol, zein and xylan, and the nanoparticles have a particle size of less than 500 nm.

[0009] The resveratrol nano-formulation is prepared by the following method: resveratrol and zein are dissolved in an organic solvent as the organic phase, xylan is dissolved in water as the aqueous phase, the organic phase is added dropwise to the aqueous phase under stirring, and the organic solvent is evaporated.

[0010] Preferably, in the preparation method of the formulation, the resveratrol is 0.5-5 parts by weight, the zein is 5-50 parts by weight, the organic solvent is an aqueous solution of ethanol, the xylan is 5-50 parts by weight, and the volume ratio of the organic phase to the aqueous phase is 1-5:1.

[0011] More preferably, in the preparation method of the formulation, the concentration of ethanol in the ethanol solution is 80%-85%.

[0012] In a further preferred embodiment, the preparation method of the formulation comprises 0.5 parts by weight of resveratrol, 5 parts by weight of zein, an 85% ethanol solution as the organic solvent, 5 parts by weight of xylan, a volume ratio of the organic phase to the aqueous phase of 2:1, and a concentration of zein in the organic solvent of 1 mg / mL to 10 mg / mL.

[0013] Preferably, the stirring speed is 200 rpm to 1000 rpm.

[0014] The preparation is an oral administration preparation.

[0015] Compared with the prior art, the present invention has the following advantages: (1) A resveratrol nanoformulation is provided, which helps to improve oral bioavailability; (2) A method for preparing resveratrol nanoparticles is provided, which has uniform and stable particle size and high drug encapsulation rate. Attached Figure Description

[0016] Figure 1 Particle size distribution diagram of the formulation in Example 4; Figure 2 Particle size distribution diagram of the formulation in Example 2; Figure 3 Scanning electron microscope image of the formulation in Example 2; Figure 4 Example 2: Particle size and polydispersity index changes of the formulation after storage at 4°C for different days. Detailed Implementation

[0017] The present invention will be further illustrated below through embodiments. It should be understood that the embodiments of the present invention are merely provided for illustrative purposes and are not intended to limit the present invention. Therefore, any simple modifications to the present invention based on the method of the present invention are within the scope of protection claimed by the present invention.

[0018] Example 1 Weigh 0.5 mg of resveratrol and 5 mg of zein, add 5 mL of 85% ethanol solution to dissolve them, and use this as the organic phase. Weigh 5 mg of xylan and dissolve it in 2.5 mL of water to use this as the aqueous phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring, stir for 30 min, and then use a rotary evaporator to remove the organic solvent.

[0019] Example 2 Weigh 5 mg of resveratrol and 50 mg of zein, add 5 mL of 85% ethanol solution to dissolve them, and use this as the organic phase. Weigh 50 mg of xylan and dissolve it in 2.5 mL of water to use this as the aqueous phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring, stir for 60 min, and then use a rotary evaporator to remove the organic solvent.

[0020] Example 3 Weigh 2 mg of resveratrol and 20 mg of zein, add 5 mL of 80% ethanol solution to dissolve them, and use this as the organic phase. Weigh 50 mg of xylan and dissolve it in 5 mL of water to use this as the aqueous phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring, stir for 60 min, and then remove the organic solvent using a rotary evaporator.

[0021] Example 4 Weigh 5 mg of resveratrol and 50 mg of zein, add 5 mL of 85% ethanol solution to dissolve them, and use this as the organic phase. Weigh 50 mg of xylan and dissolve it in 2.5 mL of water to use this as the aqueous phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring, stir for 60 min, and then use a rotary evaporator to remove the organic solvent.

[0022] Example 5 Weigh 0.25 mg of resveratrol and 10 mg of zein, add 2.5 mL of 80% ethanol solution to dissolve them, and use this as the organic phase. Weigh 10 mg of xylan and dissolve it in 1 mL of water to use this as the aqueous phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring, stir for 60 min, and then remove the organic solvent using a rotary evaporator.

[0023] Example 6 Weigh 0.5 mg of resveratrol and 5 mg of zein, add 6 mL of 90% ethanol solution to dissolve them, and use this as the organic phase. Weigh 4 mg of xylan and dissolve it in 1 mL of water to use this as the aqueous phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring, stir for 15 min, and then use a rotary evaporator to remove the organic solvent.

[0024] Comparative Example 1 Weigh 0.5 mg of resveratrol and 5 mg of human serum albumin, add 5 mL of 85% ethanol solution to dissolve them, and use this as the organic phase. Weigh 10 mg of xylan and dissolve it in 2.5 mL of water to use this as the aqueous phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring, stir for 30 min, and then remove the organic solvent using a rotary evaporator.

[0025] Comparative Example 2 Weigh 0.5 mg of resveratrol and dissolve it in 5 mL of 85% ethanol solution to form the organic phase. Weigh 5 mg of xylan and dissolve it in 2.5 mL of water to form the aqueous phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring and stir for 30 min. Then, use a rotary evaporator to remove the organic solvent.

[0026] Comparative Example 3 Weigh 0.5 mg of resveratrol and 5 mg of zein, add 5 mL of 85% ethanol solution to dissolve them, and use this as the organic phase. Weigh 5 mg of sucrose and dissolve it in 2.5 mL of water to use this as the aqueous phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring, stir for 30 min, and then use a rotary evaporator to remove the organic solvent.

[0027] Comparative Example 4 Dissolve 5 mg of zein in 5 mL of 50% ethanol solution to form the organic phase. Weigh 5 mg of xylan and dissolve it in 2.5 mL of water to form the aqueous phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring and stir for 30 min. Then, use a rotary evaporator to remove the organic solvent.

[0028] Comparative Example 5 Weigh 0.5 mg of resveratrol and 5 mg of zein, add 5 mL of dichloromethane solution to dissolve them, and use this as the organic phase. Weigh 5 mg of xylan and dissolve it in 2.5 mL of water to use this as the aqueous phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring, stir for 30 min, and then remove the organic solvent using a rotary evaporator.

[0029] Comparative Example 6 Weigh 0.5 mg of paclitaxel and 5 mg of zein, add 5 mL of 85% ethanol solution to dissolve them, and use this as the organic phase. Weigh 5 mg of xylan and dissolve it in 2.5 mL of water to use this as the aqueous phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring, stir for 30 min, and then use a rotary evaporator to remove the organic solvent.

[0030] Comparative Example 7 Weigh 5 mg of resveratrol and 0.5 mg of zein, add 5 mL of 80% ethanol solution to dissolve them, and use this as the organic phase. Weigh 5 mg of xylan and dissolve it in 1 mL of water to use this as the aqueous phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring, stir for 30 min, and then remove the organic solvent using a rotary evaporator.

[0031] Verification of Examples 1. The solutions obtained in Examples 1-6 and Comparative Examples 1-7 were filtered using a 0.45-micron microporous membrane. The particle size and polydispersity index of the resulting liquids were monitored using a laser particle size analyzer.

[0032] Table 1. Particle size and polydispersity index (PDI) of each embodiment The results are as follows Figure 1-2 As shown in Table 1, the nanoparticles prepared in the embodiments of the present invention have a normal distribution, a particle size of less than 300 nm, a PDI of less than 0.3, and uniform particle size.

[0033] 3. After adding mannitol, the freeze-drying protectant, to the solution obtained in Example 2 and mixing well, the solution was filtered through a 0.45-micron microporous membrane. The resulting liquid was then prepared into freeze-dried powder, and its morphology was observed using a scanning electron microscope.

[0034] The results are as follows Figure 3 As shown, the nano-formulation prepared in the embodiments of the present invention is approximately spherical with relatively uniform particle size.

[0035] 4. The solutions obtained in Examples 1-6 were filtered through a 0.45-micron microporous membrane. 200 μL of the resulting liquid was diluted 5 times with methanol to break the emulsion. The resveratrol drug content was detected at 306 nm using a UV spectrophotometer, and the encapsulation rate of resveratrol in the nano-formulation was calculated.

[0036] Table 2. Encapsulation efficiency of the examples and comparative examples The results are shown in Table 2. The resveratrol preparation prepared in this invention has a high encapsulation rate.

[0037] 5. The formulation prepared in Example 2 was placed at 4°C, and its particle size and polydispersity index were monitored by a laser particle size analyzer on days 1, 3, 5, 8, and 12.

[0038] The results are as follows Figure 4 As shown, the resveratrol nanoparticles prepared in the embodiments of the present invention exhibited relatively stable particle size and polydispersity index under 4°C conditions for 1-12 days.

[0039] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A method for preparing a resveratrol nanoparticle formulation, characterized in that, The nano-formulation is prepared from resveratrol, zein, and xylan, and the particle size of the nano-formulation is less than 500 nm. The formulation is prepared by the following method: resveratrol and zein are dissolved in an organic solvent as the organic phase, xylan is dissolved in water as the aqueous phase, the organic phase is added dropwise to the aqueous phase under stirring, and the organic solvent is evaporated. In the preparation method of the formulation, the weight parts of resveratrol are 0.5-5 parts, the weight parts of zein are 5-50 parts, the organic solvent is an aqueous solution of ethanol, the weight parts of xylan are 5-50 parts, and the volume ratio of the organic phase to the aqueous phase is 1-5:1; the concentration of ethanol in the ethanol solution is 80%-85%.

2. The method for preparing resveratrol nanoparticles as described in claim 1, characterized in that, In the preparation method of the formulation, the resveratrol is 0.5 parts by weight, the zein is 5 parts by weight, the organic solvent is an 85% ethanol solution, the xylan is 5 parts by weight, the volume ratio of the organic phase to the aqueous phase is 2:1, and the concentration of the zein in the organic solvent is 1 mg / mL-10 mg / mL.

3. The method for preparing resveratrol nanoparticles as described in claim 1, characterized in that, The preparation is an oral administration preparation.

Images