8-16nm pancreatic islet targeting time sequence layered immunoregulation type 2 diabetes mellitus nano drug substance and preparation method thereof

Abstract

The present application belongs to the field of plant source nano drug preparation, aiming at the defects of single drug target, organ toxicity, blood glucose rebound, islet immune imbalance and the like of existing type 2 diabetes drugs. The present application adopts a pH 6.3±0.15 phosphate buffer system to construct a hydrogenated soybean sphingomyelin-deoxycholic acid amphoteric coupling core-shell nano carrier, and self-assemble to form an 8-16 nm islet targeting nanoparticle. The raw drug has a three-layer time sequence drug release structure: 0-2 h release of the outer layer component to improve islet microcirculation, 2-6 h release of the middle layer polysaccharide to inhibit islet inflammation and induce regulatory T cell proliferation, and 6-24 h slow release of the inner core component to regulate endogenous GLP-1 secretion, repair islet beta cells and balance peripheral T cell subsets. The present application does not have chemical hypoglycemic drugs, exogenous GLP-1 and PLGA carriers, and the raw materials are all natural substances with good safety. The whole aqueous phase preparation process can be used to prepare raw drugs, capsules and the like, and is suitable for the treatment of early and middle stage insulin resistance type 2 diabetes.

Description

Technical Field

[0001] This invention relates to the field of plant-derived nanomedicine formulation technology, specifically to a nanoparticle drug for type 2 diabetes that targets pancreatic islets, releases drugs in a time-sequential manner, and possesses immunomodulatory functions, and its preparation method. The formulation, carrier structure, particle size range, targeting technology, drug release mode, and preparation process of this invention are designed based on publicly available research findings on natural drug nanocarriers, pancreatic islet-targeting formulations, and oral sustained-release formulations both domestically and internationally. Background Technology

[0002] The mainstream clinical treatment options for type 2 diabetes include GLP-1 peptide preparations, small molecule hypoglycemic drugs, PLGA-based nanomedicines, and stem cell transplantation therapy. GLP-1 peptide preparations have limited duration of action and pose risks of adverse reactions to the pancreas and thyroid. The dosage form is mainly injection, which cannot intervene in pancreatic islet immune damage. Reference

[16] Nature Reviews Endocrinology, 2022, 18(9): 541-556. Long-term use of small molecule hypoglycemic drugs can increase the metabolic burden on the liver and kidneys, easily lead to receptor resistance, and have no pancreatic tissue repair function. Reference

[17] European Journal of Medicinal Chemistry, 2023, 252: 115341. PLGA-carrier nanomedicines degrade in vivo to produce lactic acid, which has organ-accumulating toxicity; the carrier is a single-phase encapsulation structure, which is difficult to be compatible with multipolar active components and is prone to drug leakage. Reference

[18] Biomaterials Science, 2022, 10 (12): 3211-3228. Stem cell transplantation therapy carries risks of surgical trauma, immune rejection, and tumorigenesis. Reference

[19] Cell StemCell, 2023, 30 (4): 489-505. The pathogenesis of type 2 diabetes is accompanied by an imbalance of local T cell subsets in the pancreas, and chronic low-grade immune inflammation continuously damages pancreatic β cells. Existing agents cannot simultaneously regulate local and peripheral T cell subsets in the pancreas, and blood glucose rebound is likely to occur after drug withdrawal. References

[20] Nature Medicine, 2022, 28 (7): 1421-1432;

[21] Diabetologia, 2023, 66 (3): 589-602. Summary of the Invention

[0003] 3.1 Overall Design Basis This invention employs a phosphate-buffered, hydrogenated soybean sphingomyelin-deoxycholic acid composite core-shell nanocarrier. The carrier ratio, pH range, particle size range, targeted modification, stratified drug release time, and preparation process were all designed with reference to parameters published in the literature. 3.2 Composition ratios, pharmaceutical basis, and literature sources This invention is divided into a carrier component, an outer lipid-soluble component, a middle water-soluble polysaccharide component, and a core sustained-release component. The components are arranged according to the rule that the lipid phase carries lipid-soluble substances and the aqueous phase carries water-soluble substances. 3.2.1 Carrier Components Hydrogenated soybean sphingomyelin and refined deoxycholic acid constitute an amphoteric coupling lipid phase carrier, and the ratio is referenced in reference [1]. Galactose-modified chitosan has both skeletal and targeted modification functions, and the amount added is referenced in reference [3]. PEG3350 is a surface stabilizer, and the parameters are referenced in reference [2]. Phosphate buffer salt maintains the pH of the system at 6.3±0.15 to eliminate electrostatic flocculation of components, and the parameters are referenced in reference [3]. 3.2.2 Outer lipid-soluble functional components L-arginine and total saffron glycosides are carried in the outer lipid phase of the core and shell, and the ratio range is referenced in

[12] . 3.2.3 Middle-layer water-soluble polysaccharide components Bamboo leaf polysaccharide, astragalus polysaccharide and turkey lobe polysaccharide are carried in the core-shell middle aqueous phase. The total amount of polysaccharide used is within the range of natural polysaccharide nano-drug loading. The compatibility is referenced in the literature [9]. 3.2.4 Core sustained-release functional components Total saponins from bitter melon, puerarin, taurine, and Agaricus blazei polysaccharide were carried in the core, shell, and kernel, with the ratios specified in reference

[11] . 3.2.5 Raw material purity parameters The grafting rate of galactose-modified chitosan and the purity of each extract were determined with reference to the pharmacopoeia and publicly available standards for natural pharmaceutical excipients. 3.3 Targeting properties and in vivo stability The nanoparticles have a particle size of 8–16 nm, which is the applicable particle size for oral islet-targeting nanoparticles, as referenced in [4]. The nanoparticles are modified with islet-targeting peptides to recognize islet cell surface receptors, as referenced in [5]. The dual-coupled core-shell structure remains stable in the acidic environment of the stomach and is gradually dissolved and enzymatically hydrolyzed after entering the intestine. The nanoparticles are absorbed by the gastrointestinal tract and transported through the portal vein to achieve pancreatic islet enrichment. The in vivo transport pattern is described in reference [6]. 3.4 Time-sequenced drug release and pharmacological effects 0 h–2 h, the outer layer component is released, improving pancreatic islet microcirculation, with effects based on reference

[12] . 2 h–6 h, the middle layer polysaccharide component is released, inhibiting local inflammation in the islets and inducing the proliferation of pancreatic regulatory T cells, with effects based on references [7] and [9]. 6 h–24 h, the inner core component is released, acting on intestinal L cells to promote endogenous GLP-1 secretion, repair pancreatic β cells, and balance peripheral T cell subsets, with effects based on references [8] and

[10] . This invention does not contain PLGA, exogenous GLP-1 peptides, or chemical hypoglycemic drugs. According to references

[13] and

[15] , this composition is safe for use. 3.5 Preparation process This invention employs an all-aqueous process, and the process parameters for lipid phase hydration, high-pressure homogenization, surface coating, and gradient freeze-drying all refer to publicly available standards for nano-formulations. Attached Figure Description Figure 1 This is a schematic diagram comparing the particle size distribution of nanoparticles; Figure 2 This is a schematic diagram of the organ-targeted fluorescence distribution of nanoparticles. Figure 3 This is a schematic diagram of the time-sequential, layered drug release and immune regulation mechanism of the nanoparticles in this invention. References [1] International Journal of Pharmaceutics, 2022, 621:121789 [2] Journal of Controlled Release, 2023, 357:218-232 [3] Chinese Pharmaceutical Journal, 2021, 56(12):987-994 [4] Acta Biomaterialia, 2022, 145:301-315 [5] Nature Communications, 2023, 14 (1):5219 [6] Chinese Traditional and Herbal Drugs, 2022, 47 (18):4981-4989 [7] Nature Medicine, 2022, 28 (7):1421-1432 [8] Diabetologia, 2023, 66 (3):589-602 [9] Journal of Immunology, 2021, 37 (9):761-768

[10] Diabetes Care, 2022, 45 (8): 1789-1798

[11] Chinese Journal of Pharmacology, 2022, 38 (5): 721-728

[12] Phytomedicine, 2023, 112: 154687

[13] Food and Chemical Toxicology, 2022, 168: 113297

[14] Drug Evaluation Research, 2021, 44 (6): 1201-1208

[15] Journal of Ethnopharmacology, 2023, 292: 115162.

Claims

1. A type 2 diabetes nanoparticle drug with 8–16 nm pancreatic islet-targeted time-sequential hierarchical immune regulation, characterized in that, It is composed of a carrier component, an outer layer of fat-soluble functional components, a middle layer of water-soluble polysaccharide components, and a core layer of sustained-release functional components. By weight, the carrier component includes: 4.0–7.0 parts hydrogenated soybean sphingomyelin, 2.0–4.0 parts galactose-modified chitosan, 1.2–2.5 parts refined deoxycholic acid, 1.5–3.0 parts PEG3350, and 1.5–3.0 parts phosphate buffer. The outer layer of fat-soluble functional components includes: 4.0–8.0 parts L-arginine and 2.0–3.5 parts total saffron glycosides. The middle layer of water-soluble polysaccharide components includes: 8.0–14.0 parts bamboo leaf polysaccharide, 4.0–8.0 parts astragalus polysaccharide, and 1.5–3.0 parts *Trametes versicolor* polysaccharide. The core layer of sustained-release functional components includes: 7.0–12.0 parts total bitter melon saponins. 6.0–9.0 parts of puerarin, 4.0–6.5 parts of taurine, and 1.2–2.8 parts of Agaricus blazei polysaccharide; The phosphate buffer stabilizes the pH of the system at 6.3 ± 0.15; the carrier component self-assembles into core-shell nanoparticles with a particle size of 8–16 nm; the surface of the core-shell nanoparticles is modified with pancreatic islet-targeting peptides; the active pharmaceutical ingredient has a time-release structure: the outer lipid-soluble functional component is released from 0 h to 2 h, the middle water-soluble polysaccharide component is released from 2 h to 6 h, and the core sustained-release functional component is released from 6 h to 24 h.

2. The nanopharmaceutical raw material according to claim 1, characterized in that, The hydrogenated soybean sphingomyelin and refined deoxycholic acid form an amphoteric coupling structure for co-encapsulation of multipolar active components; the PEG3350 is coated on the surface of the nanoparticles to stabilize the nanoparticle size.

3. The nanoparticle pharmaceutical ingredient according to claim 1, characterized in that, The middle layer water-soluble polysaccharide component is used to inhibit local inflammation of the pancreatic islets and induce the proliferation of pancreatic regulatory T cells; the core core sustained-release functional component is used to regulate the secretion of endogenous GLP-1 by intestinal L cells, repair pancreatic β cells, and balance peripheral T cell subsets.

4. The nanopharmaceutical raw material according to claim 1, characterized in that, The galactose-modified chitosan has a grafting rate of 12%–18%, hydrogenated soybean sphingomyelin content ≥42%, purified deoxycholic acid purity ≥92%, astragalus polysaccharide purity ≥70%, saffron total glycosides purity ≥50%, and taurine purity ≥85%.

5. A method for preparing the nanoparticle pharmaceutical ingredient according to any one of claims 1–4, characterized in that, The process includes the following steps: (1) Preparation of the lipid phase: Hydrogenated soybean sphingomyelin and refined deoxycholic acid are mixed and hydrated in the dark at 58 ℃±2 ℃ for 12 min±2 min. Total saffron glycosides are added to obtain the outer lipid phase; (2) Preparation of the aqueous phase: Buffer solution is prepared using phosphate buffer, pH is adjusted to 6.3±0.15, galactose-modified chitosan is added for swelling, and the middle water-soluble polysaccharide component and the core sustained-release functional component are added in sequence to obtain the mixed aqueous phase; (3) Homogenization: The lipid phase and aqueous phase are mixed and homogenized in sequence at 650 bar±50 bar for primary homogenization and 900 bar±50 bar for secondary closed-loop homogenization. The homogenization temperature is controlled at 18 ℃±2 ℃; (4) Coating and freeze-drying: PEG3350 is added and stirred at 300 rpm±30 rpm for 10 min±2 min to complete the surface coating; then three-stage gradient freeze-drying is performed: -35 ℃±3 Pre-freezing at -10℃, sublimation drying at -10℃±2℃, and desorption drying at 25℃±2℃, the final product has a moisture content of ≤2.8%; the entire preparation process uses an all-aqueous system and no organic solvents are added.

6. The use of the nanoparticle pharmaceutical ingredient according to any one of claims 1-4 in the preparation of a drug for treating type 2 diabetes, characterized in that, The drug is used to treat early to mid-stage insulin-resistant type 2 diabetes; the drug dosage form is active pharmaceutical ingredient, capsule, granules, or oral solution.

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