Targeted nano polymer vesicle containing FcBP ligand as well as preparation method and application of targeted nano polymer vesicle
By preparing targeted nanopolymer vesicles containing FcBP ligand, the problem of low bioavailability of non-targeted nanocarriers in oral insulin delivery is solved, and the stable transmissible transport of insulin across intestinal epithelial and long-term blood sugar reduction effect is achieved, avoiding the side effects of traditional injection methods.
Patent Information
- Application Number
- CN202510740007.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-06-04
- Publication Date
- 2025-08-15
AI Technical Summary
The existing non-targeted nanocarriers have low bioavailability in oral insulin delivery, leading to adverse reactions of long-term subcutaneous insulin injection, and traditional oral methods are prone to lead to drug degradation and inactivation.
Using biotin-affinin bridging technology, FcBP ligand modified Pluronic F127 and polylactic acid block copolymer were used to prepare targeted nanopolymer vesicles. By specifically binding to the FcRn receptor, the trans-intestinal epithelial transport of insulin is achieved and bioavailability is improved.
It achieves stable embedding of insulin and crosses the intestinal epithelial barrier, improves oral bioavailability, reduces blood sugar levels, has good biocompatibility and degradability, and avoids the side effects of traditional injection methods.
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Abstract
Description
Technical Field
[0001] The present invention belongs to the field of biomedicine and relates to a targeted polymer drug carrier. Background Art
[0002] Diabetes mellitus (DM) is a metabolic endocrine disease characterized by chronic hyperglycemia. It is divided into type 1 (T1D) and type 2 (T2D) and is one of the four major non-communicable diseases and leading causes of death worldwide. Insulin (INS) is key to the treatment of diabetes. According to the International Diabetes Federation, the total number of people with diabetes is expected to increase to 643 million by 2030 and to 783 million by 2045. Patients with advanced T1D or T2D often rely on subcutaneous insulin injections to maintain blood sugar balance. However, long-term subcutaneous insulin injection is an exogenous drug delivery method that is prone to serious adverse reactions such as peripheral hyperinsulinemia, skin necrosis, nerve damage, and hypoglycemia, resulting in poor patient compliance. Therefore, the development of noninvasive insulin delivery methods is urgently needed. Oral insulin, as a safe and economical alternative, has the advantage of being non-invasive and can effectively improve patient compliance. Oral insulin is transported across the intestinal epithelium and enters the blood circulation through the portal vein. This is closest to the metabolic pathway for insulin secretion under physiological conditions, and can avoid the occurrence of the above-mentioned side effects. However, insulin is a protein drug, and direct oral administration may cause degradation and inactivation due to gastric acid, enzymes, and first-pass metabolism. Nanodrug delivery systems (DDS) based on nanoparticles (NPs) to deliver drugs are an emerging field of nanomedicine and are also widely used in oral insulin delivery research. However, non-targeted nanocarriers still have problems such as low bioavailability of oral insulin. Researchers have used different strategies to modify the surface ligands of nanocarriers, so that after entering the gastrointestinal tract, the carriers can be recognized and bound by ligands-receptors, increasing the transport and absorption of the carriers in the intestinal epithelium, thereby further improving the bioavailability of oral insulin. FcBPs (Fc domain-binding peptides), short peptides derived from the crystallizable Fc fragment of immunoglobulin (IgG) and targeted to the neonatal Fc receptor (FcRn), are considered promising ligands for enhancing the absorption of oral insulin delivery vehicles. Because FcRn is highly expressed on the intestinal epithelium, the pH gradient across the small intestine can facilitate transepithelial transport of FcBP-liganded drug delivery vehicles. Within the intestinal lumen at a pH of 6.0-6.5, delivery vehicles can enter the cells through ligand-receptor specific binding to FcRn overexpressed on intestinal epithelial cells. After transcellular transport, the FcBP ligand on the delivery vehicle dissociates from FcRn on the basolateral side of the intestinal epithelium at a pH of 7.4, exiting the cell and entering the systemic circulation, completing transmembrane transport of insulin. The FcBP ligand is linked to the nanocarrier using a biotin-avidin bridge.This technology is very beneficial for maintaining ligand activity through non-chemical bond connection, and has the advantages of convenience, speed and versatility, and can also easily connect to a variety of other ligands. There are many forms of nanoparticles for drug delivery, including spherical micelles with core-shell structures, vesicles, rods, dendrites, etc. Among them, nanopolymer vesicles have a vesicle structure similar to a hydrophilic core and a hydrophobic bilayer similar to liposomes, but are more stable than liposomes. Therefore, nanopolymer vesicles can encapsulate hydrophilic insulin in their hydrophilic core, which not only maintains the biological activity of insulin, but also enables it to successfully overcome biological barriers such as the mucus layer and small intestinal epithelial cells. These advantages of nanopolymer vesicles are also applicable to the encapsulation and delivery of other hydrophilic drugs. Summary of the Invention
[0003] The present invention aims to design and prepare a targeted nano-polymer vesicle FcBP-F127-PLA containing an FcBP ligand and composed of Pluronic F127 and polylactic acid (PLA) blocks by using biotin-avidin bridging technology, so that it can be used as an oral delivery carrier for drugs such as insulin (INS) for the treatment of diseases such as diabetes, and at the same time provide a method for preparing the polymer vesicle. The preparation of the targeted nano-polymer vesicle FcBP-F127-PLA / INS encapsulated with insulin and the research method for its hypoglycemic effect in vivo are as follows: (1) A certain amount of Biotin-F127-PLA and PLA-F127-PLA block copolymer is dissolved in a tetrahydrofuran (THF) solution, and then the mixed solution is dispersed dropwise in an insulin solution under stirring. The organic solvent DMF is removed by stirring under a fume hood. Subsequently, the mixture is centrifuged at high speed, and the precipitate is redissolved in triple-distilled water to obtain Biotin-F127-PLA / INS. The reconstituted Biotin-F127-PLA / INS nanovesicles were then added to the Avidin / PBS solution and the FcBP / PBS solution, resulting in the FcBP-F127-PLA / INS nanopolymer vesicle solution. (2) Type 1 diabetic rat models were induced by intraperitoneal injection of alloxan. After fasting for 12 hours, the FcBP-F127-PLA / INS nanovesicle solution was orally administered to the model rats. Blood glucose levels were measured at predetermined time points between 0 and 24 hours using tail vein blood samples. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 Effects of oral administration of insulin-loaded nanovesicles on blood glucose in diabetic rats. (mean ± SD, n=5; *p<0.05) DETAILED DESCRIPTION 1. The preparation method of 10% FcBP-F127-PLA / INS loaded with insulin nanovesicles with a molar content of 10% FcBP ligand is as follows: First, 1 mg of Biotin-F127-PLA and 4 mg of PLA-F127-PLA were dissolved in 1 mL of THF and added dropwise to 5 mL of 5 mg / mL insulin solution. The mixture was stirred under a fume hood for 1-2 hours to remove the organic solvent THF. Then, the mixture was centrifuged at 24000 rpm for 4 min. o C and high-speed centrifugation for 50 min. The supernatant and precipitate were separated, and the precipitate was reconstituted with ultrapure water to prepare a 10% Biotin-F27-PLA / INS drug-loaded nanovesicle solution. Avidin / PBS (1 mL, 1 mg / mL) solution was then added and incubated with shaking for 40 min. Biotin-FcBP / PBS (0.057 mL, 1 mg / mL) solution was then added and incubated with shaking for another 40 min. This resulted in a 10% FcBP-F127-PLA / INS solution. 2. A rat model of type 1 diabetes was induced by intraperitoneal injection of alloxan. Rats with blood glucose levels >16.6 mmol / L were used in subsequent experiments. After a 12-hour fast, rats were randomly divided into three groups of five each and received oral administration of 10% FcBP-F127-PLA / INS and PLA-F127-PLA / INS (50 IU / Kg), as well as subcutaneous injection of SC Free INS (5 IU / Kg). Blood glucose levels were measured by tail vein blood at predetermined time points between 0 and 24 hours to assess the oral hypoglycemic effect of 10% FcBP-F127-PLA / INS. Figure 1 Results showed that after 6 hours of oral administration, the blood sugar level in the non-targeted PLA-F127-PLA / INS group decreased by 40%, while the blood sugar-lowering effect of the targeted 10% FcBP-F127-PLA / INS group was superior to the non-targeted group, decreasing by approximately 60%, and the blood sugar-lowering effect was maintained for at least 18 hours. Unlike the two oral administration groups, the blood sugar level in the SC Free INS group decreased rapidly within 2 hours, but then quickly rebounded, failing to achieve the effect of long-term basal blood sugar reduction and being accompanied by the risk of hypoglycemia. The FcBP-targeted nanopolymer vesicles provided by the present invention have the function of targeting the FcRn receptor and can effectively and stably encapsulate drugs (especially hydrophilic drugs), allowing them to cross the intestinal epithelial barrier, thereby improving the oral bioavailability of the drugs and having good application prospects. In addition, the drug carrier of the present invention has good biocompatibility and biodegradability.
Claims
1. A novel nanopolymer vesicle FcBP-F127-PLA containing FcBP ligand, composed of Pluronic F127 and PLA.
2. The nano-polymer vesicle according to claim 1, wherein: The preparation method of drug carrier is as follows: (1) Dissolve 1 mg of Biotin-F127-PLA and 4 mg of PLA-F127-PLA in 1 mL of THF and add dropwise to 5 mL of 5 mg / mL insulin solution. Stir for 1-2 hours under a fume hood to remove the organic solvent THF. (2) Then, stir at 24000 rpm, 4 o C for 50 min. The supernatant and precipitate were separated, and the precipitate was re-dissolved in ultrapure water to prepare a drug-loaded nanovesicle solution of 10% Biotin-F27-PLA / INS. (3) Then, avidin / PBS (1 mL, 1 mg / mL) solution was added and incubated with shaking for 40 min. Then, Biotin-FcBP / PBS (0.057 mL, 1 mg / mL) solution was added and incubated with shaking for another 40 min. Finally, a 10% FcBP-F127-PLA / INS nanopolymer vesicle solution was prepared.
3. Use of the nano-polymer vesicles according to claim 1 as an oral insulin delivery carrier.
4. The use according to claim 3, characterized in that The prepared 10% FcBP-F127-PLA / INS targeted nanovesicle preparation encapsulating insulin has an oral hypoglycemic effect.