A preparation method of a nanometer preparation of vitexin

Abstract

The application discloses a preparation method of a nanometer preparation of vitexicarpin, and belongs to the field of traditional Chinese medicine preparations. The method comprises the following steps: dissolving silk fibroin and poloxamer F127 in water as an aqueous phase, dissolving vitexicarpin in acetone as an organic phase, gradually adding the organic phase into the aqueous phase under stirring, and removing the organic solvent to obtain the nanometer preparation. The preparation method is simple, the obtained nanometer preparation has uniform particle size and good dispersity, the water solubility and oral bioavailability of the vitexicarpin are significantly improved, the mucosal adhesion of silk fibroin and the stabilizing effect of poloxamer are utilized, intestinal targeting accumulation is realized, and the nanometer preparation can be used for oral treatment of ulcerative colitis and other inflammatory bowel diseases.

Description

Technical Field

[0001] This invention belongs to the field of traditional Chinese medicine preparation technology, specifically relating to a method for preparing a Vitex trifolia flavonoid nanoparticle preparation. Background Technology

[0002] Vitexicarpin (also known as Casticin) is a compound derived from Vitex trifolia (Vitex negundo). Vitex trifolia or Vitex rotundifolia Polymethoxylated flavonoids were extracted and isolated from [the sample / organism]. Modern pharmacological studies have shown that Vitex flavonoids possess significant anti-inflammatory, antioxidant, immunomodulatory, and antitumor biological activities. Particularly in gastrointestinal disease models such as inflammatory bowel disease (e.g., ulcerative colitis, Crohn's disease) and gastritis, Vitex flavonoids can exert a good gastrointestinal protective effect by inhibiting inflammatory signaling pathways such as NF-κB and MAPK, reducing the expression of pro-inflammatory cytokines (e.g., TNF-α, IL-6, IL-1β), and scavenging excess reactive oxygen species (ROS).

[0003] However, the practical application of vitexin faces two major bottlenecks. Its water solubility is extremely poor (belonging to the class of poorly soluble flavonoids), resulting in low oral bioavailability and severely impacting its in vivo efficacy. Furthermore, the complex gastrointestinal environment, with its tendency to be degraded or inactivated by gastric acid, digestive enzymes, and gut microbiota, makes it difficult to achieve effective therapeutic concentrations at the lesion site. To address these challenges, researchers have attempted to improve the oral absorption of poorly soluble drugs using nanodelivery systems, such as liposomes, polymer micelles, and solid lipid nanoparticles. However, these carriers generally suffer from complex preparation processes, residual organic solvents, low drug loading capacity, and insufficient gastrointestinal stability.

[0004] Silk fibroin, a natural structural protein extracted from silkworm silk, possesses excellent biocompatibility, biodegradability, and mucosal adhesion, and has been widely used in drug delivery system research in recent years. Under acidic gastric conditions, silk fibroin can form a gel-like protective layer, effectively encapsulating drugs and resisting gastric enzyme degradation. Simultaneously, its abundant functional groups (amino, carboxyl, and hydroxyl groups) can interact with the intestinal mucus layer, prolonging the retention time of the formulation in the intestine. However, pure silk fibroin nanoparticles suffer from drawbacks such as uneven particle size distribution and a tendency to aggregate during long-term storage.

[0005] Poloxamer 127 is an amphiphilic triblock copolymer (PEO-PPO-PEO) that has been approved by the FDA for use in pharmaceutical formulations. Currently, there are no reports on the preparation of oral nanoparticles using silk fibroin and poloxamer 127 as co-carrier materials to jointly encapsulate vitexin. Based on this, this invention proposes a vitexin nanoparticle formulation with a simple preparation process, uniform particle size, good stability, high oral absorption efficiency, and gastrointestinal targeted accumulation capability, to address the problems of low oral bioavailability and poor therapeutic effects on gastrointestinal inflammation in existing technologies. Summary of the Invention

[0006] Given the shortcomings of existing technologies, researchers have discovered that poloxamer F127 can form a hybrid network structure with silk fibroin through hydrophobic interactions, and as a surfactant, it significantly improves the dispersibility and particle size uniformity of nanoparticles. This invention provides a method for preparing a Vitexin nanoparticle formulation. This nanoparticle formulation uses both silk fibroin and poloxamer F127 as carriers, solving the challenge of oral delivery of Vitexin.

[0007] This invention provides a method for preparing a nano-formulation of Vitex trifolia flavonoids, wherein the formulation is prepared by the following method: [The method involves]... Vitexin and poloxamer F127 were dissolved in water as the aqueous phase; Vitexin was dissolved in acetone as the organic phase; the organic phase was gradually added dropwise to the aqueous phase under stirring, and the organic solvent was evaporated to obtain the Vitexin nano-formulation.

[0008] Preferably, the formulation is prepared by the following method: dissolving silk fibroin and poloxamer F127 in water as the aqueous phase, and then... The concentration of vitexin was 0.5-5 mg / mL, and the concentration of poloxamer F127 was 1-5 mg / mL. Vitexin was dissolved in acetone as the organic phase, wherein the weight ratio of vitexin to the total weight of silk fibroin to poloxamer carrier was 0.1-1:10. The organic phase was gradually added dropwise to the aqueous phase under stirring, with the volume ratio of acetone to water being 1-3:1. After evaporating the organic solvent, vitexin nano-formulation was obtained.

[0009] More preferably, in the above preparation method, the stirring speed under the stirring conditions is 200-800 rpm.

[0010] The Vitex flavonoid nanoparticles have a particle size of less than 500 nm and a polydispersity index of less than 0.3 in water.

[0011] The preparation is for oral administration and can also be used for intravenous injection.

[0012] Compared with the prior art, the present invention has the following advantages: (1) A nano-formulation of vitexin is provided, which helps to improve the solubility of vitexin; (2) A method for preparing Vitex trifolia flavonoid nanoparticles is provided, which has uniform and stable particle size and high drug encapsulation rate. Attached Figure Description

[0013] Figure 1 Particle size distribution diagram of the formulation in Example 1; Figure 2 Particle size distribution diagram of the formulation in Example 2; Figure 3 Transmission electron microscope image of the formulation in Example 4 (scale bar = 200 nm). Detailed Implementation 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.

[0014] Example 1 Weigh 0.5 mg of silk fibroin and 1 mg of poloxamer F127, add 1 mL of deionized water to dissolve them, and use this as the aqueous phase. Separately weigh 0.1 mg of vitexin and dissolve it in 1 mL of acetone to use this as the organic phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring at 200 rpm, and continue stirring to evaporate the organic solvent.

[0015] Example 2 Weigh 5 mg of silk fibroin and 5 mg of poloxamer F127, add 1 mL of deionized water to dissolve them, and use this as the aqueous phase. Separately weigh 0.5 mg of vitexin and dissolve it in 2 mL of acetone to use this as the organic phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring at 600 rpm, and continue stirring to evaporate the organic solvent.

[0016] Example 3 Weigh 20 mg of silk fibroin and 50 mg of poloxamer F127, add 10 mL of deionized water to dissolve them, and use this as the aqueous phase. Separately weigh 1 mg of vitexin and dissolve it in 20 mL of acetone to use this as the organic phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring at 800 rpm, and continue stirring to evaporate the organic solvent.

[0017] Example 4 Weigh 1 mg of silk fibroin and 5 mg of poloxamer F127, add 1 mL of deionized water to dissolve them, and use this as the aqueous phase. Separately weigh 0.5 mg of vitexin and dissolve it in 3 mL of acetone to use this as the organic phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring at 300 rpm, and continue stirring to evaporate the organic solvent.

[0018] Comparative Example 1 Weigh 20 mg of human serum albumin and 20 mg of poloxamer F127, dissolve them in 10 mL of deionized water to form the aqueous phase; separately weigh 1 mg of vitexin and dissolve it in 20 mL of acetone to form the organic phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring at 500 rpm, and continue stirring to evaporate the organic solvent.

[0019] Comparative Example 2 Weigh 2 mg of silk fibroin and 5 mg of Tween 80, add 1 mL of deionized water to dissolve them, and use this as the aqueous phase. Separately weigh 0.5 mg of vitexin and dissolve it in 20 mL of acetone to use this as the organic phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring at 500 rpm, and continue stirring to evaporate the organic solvent.

[0020] Comparative Example 3 Weigh 0.5 mg of silk fibroin and 5 mg of poloxamer F127, add 1 mL of deionized water to dissolve them, and use this as the aqueous phase. Separately weigh 1 mg of paclitaxel and dissolve it in 1 mL of acetone to use this as the organic phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring at 200 rpm, and continue stirring to evaporate the organic solvent.

[0021] Comparative Example 4 Weigh 5 mg of silk fibroin and 5 mg of poloxamer F127, add 1 mL of deionized water to dissolve them, and use this as the aqueous phase. Separately weigh 1 mg of vitexin and dissolve it in 2 mL of methanol to use this as the organic phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring at 600 rpm, and continue stirring to evaporate the organic solvent.

[0022] Comparative Example 5 Weigh 10 mg of silk fibroin and 5 mg of poloxamer F127, add 1 mL of deionized water to dissolve them, and use this as the aqueous phase. Separately weigh 1 mg of vitexin and dissolve it in 2 mL of acetone to use this as the organic phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring at 600 rpm, and continue stirring to evaporate the organic solvent.

[0023] Comparative Example 6 Weigh 10 mg of silk fibroin and 5 mg of poloxamer F127, add 2 mL of deionized water to dissolve them, and use this as the aqueous phase. Separately weigh 1 mg of vitexin and dissolve it in 0.5 mL of acetone to use this as the organic phase. Add the organic phase dropwise to the aqueous phase under magnetic stirring at 1600 rpm, and continue stirring to evaporate the organic solvent.

[0024] Verification Implementation Examples 1. The solutions obtained in the examples and comparative examples were filtered through a 0.45-micron microporous membrane. The particle size and polydispersity index of the resulting liquids were monitored using a laser particle size analyzer.

[0025] Table 1. Particle size and polydispersity index (PDI) of each embodiment The results are as follows Figure 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.

[0026] 3. The solution obtained in Example 4 was filtered through a 0.45-micrometer microporous membrane, and the resulting liquid was negatively stained with phosphotungstic acid. The morphology was observed using a transmission electron microscope.

[0027] The results are as follows Figure 3 The nano-formulation prepared in the embodiments of the present invention shown is approximately spherical with relatively uniform particle size.

[0028] 4. The solutions obtained in the examples and comparative examples were filtered through a 0.45-micron microporous membrane. 200 μL of the resulting liquid was diluted 5-fold with methanol to demulsify. The content of Vitexin was determined using a UV spectrophotometer at a wavelength of 256 nm. The encapsulation efficiency of the drug was calculated.

[0029] Table 2. Encapsulation efficiency of the examples and comparative examples The results are shown in Table 2. The formulation Vitex flavonoids prepared in this invention has a high encapsulation rate.

[0030] 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 Vitex trifolia flavonoid nanoparticle formulation, characterized in that, The formulation is prepared using the following method: silk fibroin and poloxamer F127 are dissolved in water as the aqueous phase, with a silk fibroin concentration of 0.5-5 mg / mL and a poloxamer F127 concentration of 1-5 mg / mL; vitexin is dissolved in acetone as the organic phase, wherein the weight ratio of vitexin to the total weight of silk fibroin and poloxamer carrier is 0.1-1:10; the organic phase is gradually added dropwise to the aqueous phase under stirring conditions, with a stirring speed of 200-800 rpm and a volume ratio of acetone to water of 1-3:1; the organic solvent is then evaporated to obtain the vitexin nano-formulation, wherein the particle size of the vitexin nano-formulation is less than 500 nm and the polydispersity index in water is less than 0.3.

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