Polysaccharide composition, NANO polysaccharide, preparation method therefor, and use thereof

Abstract

A polysaccharide composition, a nano polysaccharide, a preparation method therefor, and use thereof. The polysaccharide composition comprises a polysaccharide and a lipid material. The weight ratio of the polysaccharide to the lipid material is 2-50:1.

Description

Polysaccharide composition, nano polysaccharide, and preparation method and application thereof Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to a polysaccharide composition, nano polysaccharide, and a preparation method and application thereof. Background Art

[0002] Polysaccharide is an important biopolymer composed of more than 10 monosaccharide molecules dehydrated and condensed and connected by glycosidic bonds. Its structure can be described by the general formula (C6H 10 O5)n indicates that the glycosidic bonds are typically (1→3), (1→4), and (1→6)-, as well as α and β configurations. Polysaccharides typically contain more than 100 monosaccharides, with most containing tens of thousands or even millions, making them complex and large biomacromolecules.

[0003] Polysaccharides are essential building blocks of living organisms. They are abundant in nature and widely available, found in nearly all plants (ginseng, astragalus, and wolfberry), animals (heparin, chondroitin sulfate, and chitosan), fungi (shiitake mushrooms, ganoderma lucidum, and poria), microorganisms, and other organisms. Some polysaccharides, such as peptidoglycan and cellulose, serve as primary components of plant and animal cell walls, while others, such as starch and glycogen, serve as important storage nutrients. In recent years, 21 polysaccharide drugs, encompassing six categories, have been marketed in China. This vast array of polysaccharides stems from their diverse structure, including the number, type, linkage order, position, and number of sugar chains within the monosaccharide composition. This structural diversity, in turn, dictates the diversity of their pharmacological activities. Pharmacological and clinical studies have revealed hundreds of polysaccharides exhibiting diverse activities, including immune regulation, anti-tumor, lipid-lowering, hypoglycemic, anti-inflammatory, antibacterial, antioxidant, antiviral, and radiation protection. At the same time, plant polysaccharides can improve food flavor, assist enzyme catalysis processes, and reduce the toxicity, teratogenicity and potential carcinogenicity of synthetic chemicals.

[0004] Nanotechnology is an emerging field, and many nanomaterials have been used in biomedicine to prevent and treat various diseases. Nanoparticles are nanometer-sized particles. Drugs are typically encapsulated within the nanoparticle matrix or adsorbed onto the nanoparticle surface. Some drugs can also form nanoparticles themselves. Nanodrugs can exceed 100 nm, but are typically smaller than 500 nm. Due to the unique physicochemical and biological properties of nanoparticles, encapsulated drugs can more easily penetrate blood vessels and enter the body's circulation, even crossing biological barriers such as the blood-brain barrier. They have strong biopermeability, facilitating absorption. In addition, by varying the particle size of nanoparticles or modifying their surface, active or passive targeted drug delivery can be achieved, as well as sustained release effects and improved bioavailability. Nanoparticle formulations can be administered via a variety of routes, including oral, parenteral, transdermal, pulmonary, and intraocular administration. Appropriate nanostructuring methods can generally improve drug activity and bioavailability while reducing toxic side effects. Consequently, nanotechnology is increasingly being applied to the polysaccharide field.

[0005] Current research on polysaccharide nanoparticles focuses on the following areas: 1) synthesizing polysaccharide nanoselenium by complexing with selenium through methods such as reactions with selenious acid solutions; 2) forming silver nanoparticles by reacting polysaccharides with reduced silver nitrate, or hybridizing with metal elements such as gold to form gold nanoparticles; 3) chelating with metal ions and polyphenols; 4) forming nanoscale complexes with proteins or peptides, which can then be loaded with drugs; 5) hydrophilic polysaccharides are hydrophobically modified with fatty acids to become amphiphilic polysaccharides, which can be used to load poorly soluble drugs into micelles or nanoparticles, just like other amphiphilic polymers; and 6) combining positively or negatively charged polysaccharides with oppositely charged small molecules (called ionic crosslinkers) through electrostatic adsorption and ionic bridges to form polysaccharide nanodelivery carriers, which are then coupled with the drug to be loaded to form polysaccharide nanodelivery systems, such as chitosan with sodium tripolyphosphate and sodium alginate with calcium ions. All of these methods use polysaccharides as carrier materials, acting merely as excipients and not contributing to the delivery or efficacy of the active polysaccharide.

[0006] Compared to polysaccharides used as excipients (such as chitosan and sodium alginate), many polysaccharides possess clear pharmacological activities and demonstrated therapeutic effects against diseases, making them functionally active polysaccharides. Due to their complex structure, high molecular weight, and strong water solubility, polysaccharides exhibit stretching chains and a wide variety of conformations in aqueous solutions. Orally administered polysaccharides are difficult to cross the intestinal epithelial barrier and enter the bloodstream. Once in the bloodstream, they are easily phagocytosed by the reticuloendothelial system, including the liver and spleen, resulting in rapid elimination and a short retention time in the body. Furthermore, their strong hydrophilicity hinders cellular uptake. Understanding the structure-activity relationship of active polysaccharides is currently nearly complete. When used as carriers, polysaccharides can undergo various structural modifications to enhance drug loading, such as grafting or modifying hydrophobic groups or molecules (such as stearoyl) onto surface functional groups to impart amphiphilic properties and enhance assembly capacity. However, when polysaccharides are used as active ingredients for interventions or treatments in health and disease, any chemical modification may affect or even alter their activity, thereby introducing unpredictable risks in their application. Because active polysaccharides often interact with the body, cells, and targets through their surface functional groups to produce physiological effects, chemical modification of polysaccharide macromolecules is difficult to control, resulting in large batch-to-batch variability and high costs for subsequent separation and purification.

[0007] Currently, there is limited research on drug delivery systems designed to encapsulate or design polysaccharides as active drugs. The few studies that do exist have primarily encapsulated polysaccharides as water-soluble drugs using existing formulation technologies. For example, ion gelation has been used to encapsulate Ganoderma lucidum polysaccharides within chitosan carrier materials; polylactic-co-glycolic acid (PLGA) has been used as a carrier material, and solutions of Chinese yam polysaccharides and Angelica sinensis polysaccharides have been encapsulated as the inner aqueous phase within a W / O / W emulsion or the core of a PLGA nanocapsule using a double emulsion method; or polysaccharide solutions have been encapsulated within the inner aqueous phase of liposomes. However, these methods all suffer from complex preparation processes, low encapsulation efficiencies, and low drug loading, making them difficult to implement in practice. Other approaches have used polysaccharide solutions as the inner aqueous phase to prepare oil-in-water emulsions, but these face significant limitations in practical and clinical applications.

[0008] In summary, nanofiberization has the potential to enhance the activity of polysaccharides. However, to date, no nanofiberization technology for active polysaccharides with practical application value (e.g., high drug loading capacity and simple preparation) has been reported, nor have any corresponding products been developed. Therefore, providing an active polysaccharide composition, nanopolysaccharide, and preparation method thereof is a technical challenge urgently needed to be addressed by those skilled in the art. Summary of the Invention

[0009] In view of this, the present invention discloses a polysaccharide composition, a nano-active polysaccharide, and a preparation method and application thereof.

[0010] Although polysaccharides are generally hydrophilic macromolecules, there should still be some relatively hydrophobic areas within them. Monosaccharides are connected through different glycosidic bond types (α- and β-) and bonding methods, forming many types of natural polysaccharides with complex structures. Polysaccharide chains are rich in hydroxyl groups, and they can easily self-assemble through amino bond interactions within and between molecules to form special structures, such as the triple helical structure of polysaccharides such as wolfberry and carob polysaccharides. There are three crystal forms of amylose. Types A and B exhibit regular left-handed double helical structures, which are converted into single helical structures in dimethyl sulfoxide (DMSO). Type V amylose exists as a left-handed single helical structure. The polymorphic structure of starch has the same helical structure. Every six glucose units form a helix with a diameter of 1.3nm and a tilt distance of 0.8nm. Based on the size analysis, it is speculated that it can just accommodate a fat chain segment.

[0011] Combined with the examples of the present invention, it is speculated that the vast majority of active polysaccharides have similar helical structures in their molecules to varying degrees, thereby forming hydrophobic cavities or hydrophobic regions. When lipid adjuvants encounter these hydrophobic regions, the hydrophobic effect drives the interaction between the lipid adjuvant and these hydrophobic regions, or / and the host-guest interaction between the helical cavity and the fatty chain segment of the lipid adjuvant, inducing adaptive adjustment of the polysaccharide molecular conformation, spontaneously undergoing a disordered to ordered transition, and under the bridging effect of the hydrophobic region, multiple polysaccharide molecules form relatively stable nanopolysaccharides.

[0012] In order to achieve the above object, the present invention adopts the following technical solutions:

[0013] A polysaccharide composition, comprising polysaccharide and lipid material; wherein,

[0014] The polysaccharides include animal polysaccharides, plant polysaccharides, marine polysaccharides, fungal polysaccharides and bacterial polysaccharides; the polysaccharides can be pure polysaccharides with a very narrow molecular weight distribution, or total polysaccharides containing different molecular weights, or complex polysaccharides formed by mixing different polysaccharides (including those from different sources).

[0015] The lipid material is selected from the group consisting of medium-chain and long-chain fatty alcohols (C6-C50) and esters and ethers thereof, medium-chain and long-chain fatty amines (C6-C50) and amides formed with acids, medium-chain and long-chain fatty acids (C6-C50) and esters and derivatives thereof, amides of medium-chain and long-chain fatty acids (C6-C50) and derivatives thereof, amphiphilic materials containing medium-chain and long-chain fatty chains (C6-C50), long-chain fat-soluble materials with a molecular weight of less than 10,000 and amphiphilic polymers with them as lipophilic segments, terpenoids with side chain fatty chains and glycosides thereof, other small molecules with fatty chains and amphiphilic molecules with them as lipophilic parts, and the fatty chains of the lipid material are saturated fatty chains or contain one, two or more unsaturated bonds;

[0016] The weight ratio of the polysaccharide to the lipid material is 2-50:1.

[0017] Furthermore, the weight ratio of the polysaccharide to the lipid material is 3-30:1.

[0018] Furthermore, the lipid material is selected from medium-chain and long-chain fatty alcohols (C8-C30) and their esters and ethers, medium-chain and long-chain fatty amines (C8-C30) and amides formed with acids, medium-chain and long-chain fatty acids (C6-30) and their esters and derivatives, amides of medium-chain and long-chain fatty acids (C8-C30) and their derivatives, amphiphilic materials containing medium-chain and long-chain fatty chains (C8-C30), long-chain fat-soluble materials with a molecular weight of <5000 and amphiphilic polymers with them as lipophilic segments.

[0019] Furthermore, the lipid material is selected from medium-chain and long-chain fatty alcohols (C8-C18) and their esters and ethers, medium-chain and long-chain fatty amines (C8-C18) and amides formed with acids, medium-chain and long-chain fatty acids (C8-C18) and their esters and derivatives, amides of medium-chain and long-chain fatty acids (C8-C18) and their derivatives, amphiphilic materials containing medium-chain and long-chain fatty chains (C8-C18), long-chain fat-soluble materials with a molecular weight of <3000 and amphiphilic polymers with them as lipophilic segments.

[0020] It is worth mentioning that during the research process, the inventor unexpectedly discovered that under the induction of certain lipid adjuvants, polysaccharide macromolecules can self-assemble into nanostructures with good stability and can be administered orally and intravenously, and then conducted in-depth and systematic research on it. The results showed that many lipid materials can induce polysaccharide macromolecules to assemble into nanoaggregates (nanopolysaccharides) in aqueous solution, including medium-chain and long-chain fatty alcohols (C6-C50) and their esters and ethers, medium-chain and long-chain fatty amines (C6-C50) and amides formed with acids, medium-chain and long-chain fatty acids (C6-C50) and their esters and derivatives, amides of medium-chain and long-chain fatty acids (C6-C50) and their derivatives, amphiphilic materials containing medium-chain and long-chain fatty chains (C6-C50), and materials with molecular weight <1000. 0 (including PLA, PLGA, PCL) and amphiphilic polymers with them as lipophilic segments; terpenoids with side fatty chains (no less than 5 carbon atoms) (cholesterol, cholesterol, protopanaxadiol, protopanaxatriol, etc.) and their glycosides (such as ginsenoside Rb1 and ginsenoside Rg1); other small molecules with fatty chains (no less than 5 carbon atoms, including carbon atoms across ester bonds) (such as vitamin E) and amphiphilic molecules with them as lipophilic moieties (such as TPGS). The fatty chains of these lipid materials can be saturated or contain one, two, or more unsaturated bonds.

[0021] The types of polysaccharides suitable for inducing the formation of nanoaggregates (or nanoassemblies) by these lipid adjuvants are very wide, covering various active polysaccharides from plants, animals, fungi, etc., such as: Astragalus polysaccharide, Plantago polysaccharide, Versicolor polysaccharide, Lycium barbarum polysaccharide, soybean polysaccharide, Lentinus edodes polysaccharide, Angelica polysaccharide, Polyporus polysaccharide, Tremella fuciformis polysaccharide, Aloe polysaccharide, Chinese yam polysaccharide, Fritillaria polysaccharide, Cordyceps polysaccharide, Eucommia polysaccharide, Achyranthes bidentata polysaccharide, Polygonatum sibiricum polysaccharide, Bletilla striata polysaccharide, Ginseng polysaccharide, Gardenia jasminoides polysaccharide, Dendrobium polysaccharide, Lilium polysaccharide, etc., and lipid adjuvants of different proportions can all induce the formation of these polysaccharide nanoassemblies. Whether it is a pure polysaccharide with a very narrow molecular weight distribution, or a total polysaccharide containing different molecular weights, or a complex polysaccharide formed by mixing different polysaccharides (including different sources), lipid adjuvants can all induce the formation of these polysaccharide nanoassemblies. Of course, different polysaccharides may have their own most suitable lipid inducer and the most suitable polysaccharide-excipient ratio. The combination of different lipid adjuvants may also be better than the preparation effect of a single lipid adjuvant.

[0022] Furthermore, the polysaccharide composition also includes pharmaceutically acceptable excipients, which include commonly used additives for injections (pH regulators, surfactants, suspending agents, additives for delaying drug release, isotonicity regulators, local analgesics, antibacterial agents), adhesives, disintegrants, lubricants, tablet fillers / diluents, and tablet lubricants.

[0023] The second object of the present invention is to provide a nano polysaccharide prepared from the polysaccharide composition described above.

[0024] In order to achieve the above object, the present invention adopts the following technical solutions:

[0025] A nano polysaccharide is prepared by dissolving a lipid material in a water-miscible organic solvent, adding the solution to a polysaccharide solution under stirring or ultrasound to induce self-assembly of the polysaccharide, and then removing the organic solvent to allow the lipid material and the polysaccharide to assemble into nanoparticles with a particle size of 20-1000 nm.

[0026] It should be noted that the nano-polysaccharide can change the way polysaccharides interact with cells, pharmacokinetics and tissue distribution behavior, improve the cell uptake of polysaccharides, delay the clearance of polysaccharides from the blood circulation, and prolong the retention time in the body, thereby significantly improving the in vitro and in vivo activity and efficacy of active polysaccharides.

[0027] Furthermore, the preparation method of the nano polysaccharide specifically comprises the following steps:

[0028] 1) dissolving the lipid material in ethanol, methanol, acetone, isopropanol or a mixed solvent thereof to obtain a lipid material solution, and dissolving the polysaccharide in water to obtain a polysaccharide solution;

[0029] 2) adding the lipid material solution to the polysaccharide solution under stirring or ultrasound, and evaporating the solvent to obtain nano-polysaccharides with an average particle size of 20-1000 nm;

[0030] 3) If the particle size of the obtained nanopolysaccharide is too large or the particle size distribution is too wide, the particle size and / or particle size distribution can be further reduced by high-pressure homogenization;

[0031] 4) The obtained nano-polysaccharide can be further spray-dried or freeze-dried to become a solid powder to suit different uses.

[0032] Furthermore, the evaporation in step 2) is one of natural evaporation, heating evaporation, and reduced pressure evaporation, or a combination thereof, and the homogenization pressure in step 3) is 500-4000 bar.

[0033] The present invention also provides a second technical solution for preparing nano-polysaccharides using amphiphilic lipid materials. The amphiphilic lipid material is directly added to an aqueous solution of polysaccharide, stirred to hydrate it, and then ultrasonicated (and optionally supplemented with heating) to accelerate its diffusion into or between polysaccharide molecules, promoting its interaction with the polysaccharide molecules and inducing the polysaccharide molecules to assemble into nano-aggregates. This technical solution has the advantage of not using organic solvents. However, its disadvantages are that the preparation process is slightly slower, the resulting nano-polysaccharide particle size is slightly larger than that of the first technical solution, and the particle size distribution is also slightly wider. However, high-pressure homogenization can more effectively reduce the particle size and particle size distribution of the nano-polysaccharide.

[0034] The present invention also seeks to protect the use of the nano polysaccharide in the preparation of food, health products and pharmaceutical products.

[0035] Moreover, the nano polysaccharide can be further loaded with fat-soluble small molecules or proteins, polypeptides, and nucleic acid macromolecules to become nanoparticles with health-care and therapeutic effects, thereby achieving in vivo delivery of poorly soluble drugs and macromolecular drugs and synergistic enhancement based on the nano effect, or synergistic enhancement with polysaccharides; and the application routes of the nano polysaccharide include oral, injection, mucosal, cavity, wound or external use.

[0036] Furthermore, the nanopolysaccharide is used in foods, health products and medicines for anti-inflammatory, anti-oxidation, (auxiliary) anti-tumor, (auxiliary) anti-cardiovascular and cerebrovascular disease, neuroprotection, (auxiliary) anti-fibrosis, lipid-lowering, blood pressure-lowering, blood sugar-lowering, anti-ADHD, anti-scarring and the like.

[0037] Compared with the prior art, the present invention discloses a polysaccharide composition, a nano-polysaccharide, and a preparation method and application thereof, wherein active polysaccharide nanoparticles are prepared by inducing hydrophobic interactions with lipid chaperone molecules, and has the following significant advantages:

[0038] 1) The excipients are inexpensive and safe - the liposome adjuvants selected are mostly physiologically inert, and many are commonly used pharmaceutical excipients, which are inexpensive and safe. 2) The preparation process is extremely simple and convenient for industrial production - the alcohol solution of the lipid adjuvant is added to the polysaccharide solution under ultrasound or stirring, and the organic solvent is removed (high-pressure homogenization can be used to reduce the particle size if necessary). 3) Ultra-high polysaccharide drug loading - only a small amount of lipid material is used in the preparation process, and the main body of the nano polysaccharide is the polysaccharide. 4) The structure of the polysaccharide is not changed, and the pharmacological activity of the polysaccharide can be preserved intact - the present invention induces the nano assembly of polysaccharides through the hydrophobic interaction of the added lipid adjuvant. The formation of nano polysaccharides relies solely on physical interactions and does not cause any changes to the chemical structure of the polysaccharide, thereby not affecting its activity. 5) Significantly improve the activity of polysaccharides. Nano-polysaccharides significantly increase the uptake of polysaccharides by tissue cells (by more than 5 times), thereby significantly improving the in vitro activity of polysaccharides. 6) Significantly improve pharmacokinetic behavior. The AUC of nano-carob polysaccharide in the blood is higher than that of free polysaccharide, significantly prolonging its retention in the body. 7) Significantly improve the in vivo efficacy - Nano-polysaccharides can increase the uptake of tissue cells and prolong their retention in the body. Whether administered orally or intravenously, nano-polysaccharides have significantly improved their efficacy compared to free polysaccharides. 8) It is expected to significantly improve the absorption and oral bioavailability of polysaccharides. After many small molecule drugs are prepared into nanoparticles, their oral absorption is improved. The in vivo efficacy of nano-polysaccharides administered orally is significantly better than that of polysaccharide solutions, suggesting that nano-polysaccharides are expected to significantly improve the oral absorption and bioavailability of polysaccharides. BRIEF DESCRIPTION OF THE DRAWINGS

[0039] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are merely embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying any creative work.

[0040] Figure 1 is a photo of the particle size and particle size distribution of nanocarob polysaccharide CSPS@OANPS (polysaccharide-oleic acid = 15:1).

[0041] Figure 2 is a transmission electron microscope photo of carob nanopolysaccharide.

[0042] FIG3 is a curve showing the change of particle size and polydispersity index of carob nanopolysaccharide over time during storage at room temperature.

[0043] FIG4 is a curve showing the change of particle size and polydispersity index of nanocarob polysaccharide over time during incubation at 37° C. in different physiological media.

[0044] Figure 5 shows the effects of different concentrations of free polysaccharide (A) and nano-polysaccharide (B) on the survival rate of RAW 264.7 macrophages, as well as the effects of free polysaccharide and nano-polysaccharide on the release of NO (C), IL-6 (D), IL-10 (E), and TNF-a (F) after 48 hours of treatment with RAW 264.7 macrophages (n=3). (****P<0.0001 vs. control group; # P < 0.05; ## P < 0.01; ### P < 0.001; #### P<0.01vs.LPS group; && P<0.01).

[0045] Figure 6 shows the cough latency (A), the number of coughs within 2 minutes (B), the levels of NO (C), IL-6 (D), IL-10 (E), and TNF-a (F) in the lungs of mice with lipopolysaccharide (LPS) and ammonia-induced lung inflammation-cough model after oral and intravenous injection of free polysaccharide and nano-polysaccharide (n=10), and lung HE pathological sections (G). (****P<0.0001 vs. control group; # P < 0.05; ## P < 0.01; ### P < 0.001; #### P<0.01vs.LPS group; && P<0.01).

[0046] Figure 7 shows the fluorescence images of Caco-2 cells taking up FITC-labeled free polysaccharides (A) and nano-polysaccharides (B) and their semi-quantitative analysis (C). The blue color represents 4,6-diamidino-2-phenylindole (DAPI), and the green color represents FITC-labeled free polysaccharides or nano-polysaccharides.

[0047] FIG8 is a curve showing the change in blood drug concentration over time after intravenous injection of IR783-labeled free polysaccharide and nano-polysaccharide in mice (n=3).

[0048] Figure 9 is a comparison of the Congo red test results of carob polysaccharide CSPS and nanocarob polysaccharide CSPS@OANPS.

[0049] Figure 10 is a comparison of the FT-IR spectra of carob polysaccharide CSPS and nanocarob polysaccharide CSPS@OANPS. DETAILED DESCRIPTION

[0050] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0051] The term "embodiment" is used herein specifically to describe any embodiment as "exemplary," and should not be construed as superior or preferable to other embodiments. Performance indicators in the embodiments of this application were tested using conventional testing methods in the art, unless otherwise specified. It should be understood that the terms used in this application are intended solely to describe specific implementations and are not intended to limit the disclosure herein.

[0052] Unless otherwise specified, the technical and scientific terms used herein have the same meanings as commonly understood by ordinary technicians in the technical field to which this application belongs; other experimental methods and technical means not specifically specified in this application refer to experimental methods and technical means commonly used by ordinary technicians in this field.

[0053] In order to better illustrate the content of this application, numerous specific details are provided in the specific examples below. It should be understood by those skilled in the art that this application can be implemented without certain specific details. In the examples, some methods, means, instruments, equipment, etc. well known to those skilled in the art are not described in detail in order to highlight the main purpose of this application.

[0054] Under the premise of no conflict, the technical features disclosed in the embodiments of this application can be combined arbitrarily, and the resulting technical solutions belong to the contents disclosed in the embodiments of this application.

[0055] For a better understanding of the present invention, the present invention is further specifically described below through the following examples, but it should not be understood as limiting the present invention. Some non-essential improvements and adjustments made by those skilled in the art based on the above invention content are also considered to fall within the scope of protection of the present invention.

[0056] Example 1. Nano Astragalus Polysaccharide with Glycosides of Terpenoid Compounds with Side Fatty Chains as Excipients

[0057] Astragalus polysaccharide APS was accurately weighed and dissolved in water to prepare a 5 mg / mL polysaccharide solution. 10 mg of ginsenoside Rg1, ginsenoside Rb1, and total ginsenosides were weighed separately and dispersed in 10 mL of the polysaccharide solution. Ultrasound was performed at 37°C, and the particle size, PDI, and potential were measured at 5, 10, 20, 30, and 60 min. See Table-1, Table-2, and Table-3. Nano-astragalus polysaccharides were successfully prepared.

[0058] Table-1 Particle size, PDI, and potential of Rg1-APS (weight ratio 1:5) nano-astragalus polysaccharide obtained at different ultrasonic times

[0059] Table-2 Particle size, PDI, and potential of Rb1-APS (weight ratio 1:5) nano-astragalus polysaccharides obtained at different ultrasonic times

[0060] Table-3 Particle size, PDI, and potential of nano-astragalus polysaccharides of total ginsenosides-APS (weight ratio 1:5) obtained by ultrasound at different times

[0061] Example 2. Nano-astragalus polysaccharide containing fatty acids, fatty alcohols, fatty amines and their amides as excipients (polysaccharide-excipient = 5:1, weight ratio)

[0062] A 5 mg / mL aqueous solution of astragalus polysaccharide was prepared, and some of neodecanoic acid, oleic acid, linoleic acid, ricinoleic acid, linolenic acid, stearic acid, arachidic acid, octanol, lauryl alcohol, stearyl alcohol, dodecylamine, stearylamine, and stearylethanolamine were weighed to prepare a 4 mg / mL methanol solution. 0.5 mL of methanol solution (2 mg of excipients) was dripped into 2 mL of polysaccharide solution (10 mg of polysaccharide) under ultrasonic conditions, and ultrasonication was continued for 5 minutes. The organic phase was removed by rotary evaporation under reduced pressure (homogenization at 1400 bar for 10 times if necessary), and the particle size, PDI, and potential were measured. The results are shown in Table 4. Nano-polysaccharides were successfully obtained.

[0063] Table-4 Particle size, PDI, and potential of nano-astragalus polysaccharides with fatty acids, fatty alcohols, fatty amines and their amides as excipients

[0064] Example 3. Nanopolysaccharide with fatty acid ester as auxiliary material

[0065] Prepare an aqueous solution of Astragalus polysaccharide with a concentration of 5 mg / mL, weigh the monooleylglycerol Peceol TM , monolinoleylglycerol CC, Propylene glycol monolaurate Lauroglycol TM 90. Mono- and di-caprylic acid capric glyceride Labrafac MC60 (mainly composed of monoester and a small amount of diester) was prepared into a 4 mg / mL ethanol solution. 0.5 mL of ethanol solution (2 mg of excipient) was dripped into 2 mL of polysaccharide solution (10 mg of polysaccharide) under ultrasonic conditions. Ultrasonication was continued for 5 minutes. The organic phase was removed by rotary evaporation at 45°C. The particle size, PDI, and potential were measured and shown in Table 5. Nano-polysaccharides were successfully obtained.

[0066] Aqueous solutions of different polysaccharides (Polygonatum sibiricum polysaccharide, Panax ginseng polysaccharide, Gardenia jasminoides polysaccharide, Fritillaria thunbergii polysaccharide, Aloe vera polysaccharide, Dioscorea opposita polysaccharide, Astragalus membranaceus polysaccharide, Polyporus umbellatus polysaccharide, Ceratonia siliqua polysaccharide, etc.) were prepared with a concentration of 5 mg / mL. Mono- and di-caprylic acid capric glyceride Labrafac MC60 (mainly composed of monoester and a small amount of diester), lauryl laurate, amyl octanoate, and lauryl palmitoleate were weighed to prepare a 4 mg / mL ethanol solution. 0.25 mL-0.5 mL of ethanol solution was dripped into 2 mL of polysaccharide solution (10 mg of polysaccharide) under ultrasonic conditions, and ultrasonication was continued for 5 minutes. The organic phase was removed by rotary evaporation at 45°C. The particle size, PDI, and potential were measured as shown in Table 5. Nano-polysaccharides were successfully obtained.

[0067] Table-5 Particle size, PDI, and potential of nanopolysaccharides with fatty acid esters, edible oils, and fat-soluble small molecules as auxiliary materials

[0068] Example 4. Nanopolysaccharide with edible oil and fat-soluble small molecules as auxiliary materials (polysaccharide-auxiliary material = 8:1, weight ratio)

[0069] Refined soybean oil, olive oil, corn oil and other edible oils containing medium-chain and long-chain fatty acid glycerides were accurately weighed and dissolved in ethanol to prepare a 10 mg / mL solution. Astragalus polysaccharide, cordyceps polysaccharide, carob polysaccharide and the like were accurately weighed and dissolved in water to prepare a 10 mg / mL solution. 0.5 mL of ethanol solution of different edible oils (5 mg of excipients) was taken and slowly dripped into 4 mL of polysaccharide solution (40 mg of polysaccharide) under 250W water bath ultrasound. The organic solvent was removed by rotary evaporation at 45°C. The particle size, PDI and Zeta potential were tested. The results are shown in Table 6 below, showing that nano cordyceps polysaccharides were successfully produced.

[0070] 5 mg of protopanaxadiol PPD (a fat-soluble aglycone of ginsenosides), protopanaxadiol, vitamin E, cholesterol, and cholestanol were accurately weighed and dissolved in anhydrous ethanol to prepare a 10 mg / mL solution. In 0.5 mL, 50 mg of cordyceps polysaccharide, carob polysaccharide, lentinan polysaccharide, and eucommia polysaccharide were accurately weighed and dissolved in 5 mL of water as the aqueous phase. 0.5 mL of ethanol solution was slowly dripped into 4 mL of polysaccharide solution by ultrasonication. The organic solvent was removed by rotary evaporation at 45 ° C. The particle size, PDI, and Zeta potential were tested. The results are shown in Table 6 below, indicating that all nano-polysaccharides were successfully prepared.

[0071] Table-6 Particle size, PDI, and potential of nanopolysaccharides with edible oil and fat-soluble small molecules as auxiliary materials

[0072] Example 5. Nano-astragalus polysaccharide and nano-lycium barbarum polysaccharide containing amphiphilic molecules with fatty chain segments as excipients

[0073] Prepare an aqueous solution of astragalus polysaccharide with a concentration of 5 mg / mL, and weigh polyoxyethylene castor oil EL, TPGS, linoleoyl polyoxyethylene-6 glyceride, M2125CS, Caprylic / Capric Macrogol Glycerides Maize 59 and Planico P188 were prepared into ethanol solutions. Egg yolk phospholipids, soybean lecithin, dioleoylphosphatidylcholine, and DSPE-PEG2000 were separately weighed and dissolved in acetone. 0.5 mL of either ethanol solution or acetone solution was added dropwise to 2 mL of the polysaccharide solution under stirring. Ultrasonication was continued for 5 minutes, and the organic phase was removed by rotary evaporation at 45°C. Particle size, PDI, and potential were measured (see Table 7). Nanopolysaccharides were successfully obtained.

[0074] Prepare a 5 mg / mL aqueous solution of Lycium barbarum polysaccharide and take PLA 1000 -PEG 1000 、PCL 1000 -PEG 1000 , PLGA 1000 -PEG 1000 Several 4 mg / mL ethanol solutions were prepared, and 0.25 mL of ethanol solution (1 mg of excipient) was dripped into 2 mL of polysaccharide solution (10 mg of polysaccharide) under ultrasonic conditions. Ultrasonication was continued for 5 minutes, and the organic phase was removed by rotary evaporation at 45°C. The particle size, PDI, and potential were measured and shown in Table 7. Nano-polysaccharides were successfully obtained.

[0075] Table-7 Particle size, PDI, and potential of astragalus polysaccharide and wolfberry polysaccharide nanoassemblies induced by amphiphilic molecules with fatty chain segments

[0076] Example 6 Nanopolysaccharide with long-chain fat-soluble material as excipient (polysaccharide-excipient = 5:1, weight ratio)

[0077] Materials: Polylactic acid (PLA) (molecular weight 2000), polycaprolactone (PCL) (molecular weight 1000), polylactide-glycolide copolymer (PLGA) (molecular weight 500)

[0078] A polysaccharide aqueous solution with a concentration of 5 mg / mL was prepared. Some PLA, PCL, and PLGA were prepared into a 4 mg / mL ethanol solution. 0.5 mL of ethanol solution (2 mg of excipients) was taken and dripped into 2 mL of polysaccharide solution (10 mg of polysaccharide) under ultrasonic conditions. The ultrasonication was continued for 5 minutes. The organic phase was removed by rotary evaporation at 45°C. The particle size, PDI, and potential were measured and shown in Table 8. After being placed at room temperature for 2 days, the particle size, PDI, and potential were measured again. The results are shown in Table 9. Nano-polysaccharides were successfully obtained and had good stability.

[0079] Table-8 Nanopolysaccharides with PLA (MW2000), PCL (MW1000), and polylactide-glycolide copolymer PLGA (MW600) as excipients

[0080] Table-9 Particle size, particle size distribution and potential of nanopolysaccharides with PLA (MW2000), PCL (MW1000) and polylactide-glycolide copolymer PLGA (MW600) as excipients after being placed at room temperature for 2 days

[0081] Example 7 Long-chain fat-soluble material as auxiliary material nanopolysaccharide (polysaccharide-auxiliary material = 10:1, mass ratio)

[0082] A polysaccharide aqueous solution with a concentration of 5 mg / mL was prepared. Some PLA, PCL, and PLGA were prepared into 4 mg / mL ethanol solution. 0.25 mL of ethanol solution (1 mg of excipient) was taken and dripped into 2 mL of polysaccharide solution (10 mg of polysaccharide) under ultrasonic conditions. The ultrasonication was continued for 5 minutes. The organic phase was removed by rotary evaporation at 45°C. The particle size, PDI, and potential were measured and shown in Table 10. Nano-polysaccharides were successfully obtained.

[0083] Table-10 Particle size and particle size distribution of nanopolysaccharides with PLA, PCL, and poly(lactide-co-glycolide) copolymer PLGA of other molecular weights as excipients

[0084] Example 8: Labrafac MC60, Labrafil M2125CS, linoleic acid as auxiliary materials, nano-astragalus polysaccharides in different proportions

[0085] A 5 mg / mL aqueous solution of Astragalus polysaccharide APS was prepared, and Labrafac MC60, Labrafil M2125CS, and some linoleic acid were respectively prepared into 4 mg / mL ethanol solution. 0.5 mL of ethanol solution (containing 2 mg of excipients) was respectively dripped into 2 mL (containing 10 mg of polysaccharide), 4 mL (containing 20 mg of polysaccharide), 8 mL (containing 40 mg of polysaccharide), and 20 mL (containing 100 mg of polysaccharide) polysaccharide solutions under ultrasonic conditions. The ultrasonication was continued for 5 minutes, and the organic phase was removed by rotary evaporation at 45°C. The particle size, PDI, and potential were measured. The results are shown in Table 11, Table 12, and Table 13, respectively. It can be seen that nano-polysaccharides were successfully prepared.

[0086] Table 11 Particle size, PDI, and potential of nano-astragalus polysaccharides with different proportions using Labrafac MC60 as an excipient

[0087] Table-12 Particle size, PDI, and potential of nano-astragalus polysaccharides with Labrafil M2125 CS as excipient and different proportions

[0088] Table-13 Particle size, PDI, and potential of nano-astragalus polysaccharides with linoleic acid as an excipient and different proportions

[0089] The particle size was measured again after being placed at room temperature for one week. The results are shown in Table 14, Table 15 and Table 16. It can be seen that the nano-astragalus polysaccharides prepared with the three excipients have good particle size stability when placed at room temperature.

[0090] Table-14 Particle size, PDI, and potential of nano-astragalus polysaccharides with different proportions of Labrafac MC60 as excipients and different proportions after 7 days

[0091] Table-15 Particle size, PDI, and potential of nano-astragalus polysaccharides after 7 days of use with Labrafil M2125 CS as an excipient and different proportions

[0092] Table-16 Particle size, PDI, and potential of nanoparticles and astragalus polysaccharides with different ratios after 7 days of storage as an excipient of linoleic acid

[0093] Example 9: Induction of Various Nanopolysaccharides by Caprylic / Capric Medium Chain Triglycerides (Polysaccharide-Excipient = 5:1)

[0094] Prepare 5mg / mL aqueous solutions of Astragalus, Plantago, Versicolor, Lentinus edodes, Soybean, Angelica, and Lycium barbarum polysaccharides, and take caprylic and capric medium chain triglycerides Labrafac TM Lipophile WL 1349 was prepared into a 4 mg / mL methanol solution, and 0.5 mL (m 载 =2mg) was dripped into 2mL (m 药 =10 mg) polysaccharide solution, and continued ultrasonication for 5 min. The organic phase was removed by rotary evaporation at 45°C, and the particle size, PDI, and potential were measured. The results are shown in Table 17, indicating that nano-polysaccharides were successfully prepared.

[0095] After 10 times of homogenization at 1400 bar, the particle size, PDI, and potential were measured. The results are shown in Table 18. It can be seen that high-pressure homogenization can be used to reduce the average particle size and particle size distribution of the obtained nanopolysaccharide when necessary.

[0096] Table-17 Particle size, PDI, and potential of nano Astragalus polysaccharide with caprylic acid and capric acid medium chain triglyceride as excipient

[0097] Table-18 Particle size, PDI, and potential of nano-astragalus polysaccharide with medium chain triglyceride as excipient after 10 homogenizations at 1400 bar

[0098] Example 10 Various Nanopolysaccharides with Stearic Acid as Excipient

[0099] Aqueous solutions of polysaccharides from Astragalus, Plantago, Versicolor, Lentinus edodes, soybean, Angelica, and Lycium barbarum were prepared at a concentration of 5 mg / mL respectively. Some stearic acid was taken to prepare a 4 mg / mL methanol solution. 0.5 mL of methanol solution was dripped into 2 mL of polysaccharide solution under ultrasonic conditions. Ultrasonication was continued for 5 minutes. The organic phase was removed by rotary evaporation at 45°C. The particle size, PDI, and potential were measured. The results are shown in Table 19. It can be seen that nano-polysaccharides were successfully prepared.

[0100] After 10 times of homogenization at 1400 bar, the particle size, PDI, and potential were measured. The results are shown in Table 20. It can be seen that high-pressure homogenization can be used to reduce the average particle size and particle size distribution of the obtained nanopolysaccharide when necessary.

[0101] Table-19 Particle size, PDI, and potential of nanopolysaccharides with stearic acid as excipient (polysaccharide-excipient = 5:1)

[0102] Table-20 Particle size, PDI, and potential of nanoassemblies with stearic acid as an excipient after homogenization at 1404 bar for 10 times

[0103] Example 11 Nanoassemblies of Various Polysaccharides with Oleic Acid as Excipient (Polysaccharide: Excipient = 5:1)

[0104] Weigh 40 mg of polysaccharide, dissolve it in 4 ml of deionized water, and ultrasonicate it for 5 minutes to obtain a polysaccharide solution (10 mg / mL). Take 54 mg of oleic acid and dissolve it in 3 mL of acetone. Take 444 μl of each (about 8 mg in total) and drip it into 4 mL of polysaccharide solution under ultrasonic conditions. Continue ultrasonicating for 5 minutes and then remove the organic solvent by vacuum evaporation at 45°C. Homogenize at 1400 bar for 10 times, measure the particle size, PDI, and potential. The results are shown in Table 21. It can be seen that nano-polysaccharides were successfully prepared.

[0105] Table-21 Particle size, PDI, and potential of nanopolysaccharides with oleic acid as an auxiliary material

[0106] Example 12 Nanopolysaccharide loaded with protopanaxadiol PPD using TPGS as an excipient

[0107] Accurately weigh 5 mg of protopanaxadiol (PPD) and dissolve it in 0.5 mL of anhydrous ethanol. Accurately weigh 30 mg of polysaccharide and 5 mg of TPGS and dissolve them in 5 mL of water as the aqueous phase. The ethanol solution was slowly dripped into the aqueous phase using ultrasound. The organic solvent was removed by rotary evaporation at 45°C. Particle size, PDI, and zeta potential were measured. The results are shown in Table 22 below, indicating that all nanopolysaccharides were successfully produced.

[0108] Table-22 Particle size, PDI, and potential of nanopolysaccharides loaded with PPD using TPGS as an excipient

[0109] Example 13 Nano soybean polysaccharide and nano astragalus polysaccharide with TPGS as auxiliary material are further loaded with ginsenoside CK

[0110] Accurately weigh 5 mg of ginsenoside CK and dissolve it in 0.5 mL of anhydrous ethanol. Accurately weigh 30 mg of soybean polysaccharide or astragalus polysaccharide and 5 mg of TPGS and dissolve them in 5 mL of water as the aqueous phase. The ethanol solution was slowly dripped into the aqueous phase under ultrasound. The organic solvent was removed by rotary evaporation at 45°C under reduced pressure. The particle size, PDI, and zeta potential were measured. The results are shown in Table 23, indicating that nanopolysaccharides were successfully prepared and loaded with ginsenoside CK.

[0111] Table-23 Particle size, PDI, and potential of nano soybean polysaccharides and nano astragalus polysaccharides loaded with ginsenoside CK using TPGS as an excipient

[0112] Example 14 Nanopolysaccharide with oleic acid as auxiliary material loaded with ginsenoside CK (polysaccharide-CK-oleic acid = 8:1:2)

[0113] Accurately weigh 5 mg of ginsenoside CK and 10 mg of oleic acid and dissolve them in 1 mL of anhydrous ethanol. Accurately weigh 40 mg of polysaccharide and dissolve it in 5 mL of water. Slowly drop the ethanol solution into the polysaccharide solution under 150W ultrasound. Remove the ethanol by rotary evaporation under reduced pressure at 45°C. Detect the particle size, PDI, and Zeta potential. The results are shown in Table 24. It can be seen that nanopolysaccharides were successfully prepared and loaded with ginsenoside CK.

[0114] Table-24 Particle size, PDI, and potential of nanopolysaccharides loaded with PPD using oleic acid as an excipient

[0115] Example 15 Nanopolysaccharide with Oleic Acid as Excipient Loaded with Ginsenoside Rh2 (Polysaccharide-Oleic Acid-Rh2=5:2:1)

[0116] 25 mg of Rh and 10 mg of oleic acid were precisely weighed and ultrasonically dissolved in 500 μL of anhydrous ethanol. 25 mg of soybean polysaccharide was dissolved in 5 mL of water. The ethanol solution was added dropwise to the polysaccharide solution under 200W waterbath sonication. The ethanol was removed by rotary evaporation under reduced pressure at 45°C. The particle size, PDI, and zeta potential were measured. The results are shown in Table 25, indicating that all nanopolysaccharides were successfully prepared and loaded with ginsenoside Rh2.

[0117] Table-25 Particle size, PDI, and potential of nanopolysaccharides loaded with ginsenoside Rh2 after oleic acid was used as an excipient

[0118] Example 16 Nanopolysaccharide with Oleic Acid as Excipient Loaded with Ginsenoside Rh2 (Polysaccharide-Oleic Acid-Rh2=4:1:1)

[0119] 5 mg of ginsenoside Rh2 and 5 mg of oleic acid were precisely weighed and dissolved in 600 μL of anhydrous ethanol by ultrasonication. 20 mg of soybean polysaccharide was dissolved in 5 mL of water. The ethanol solution was added dropwise to the polysaccharide solution under 250W ultrasonication. The organic solvent was removed by vacuum rotary evaporation at 45°C, and the particle size, PDI, and zeta potential were measured. The results are shown in Table 26, indicating that all nanopolysaccharides were successfully prepared and loaded with ginsenoside Rh2.

[0120] Table-26 Particle size, PDI, and potential of nanopolysaccharides loaded with ginsenoside Rh2 using oleic acid as an excipient

[0121] Example 17 Nanopolysaccharide loaded with ginsenoside Rh2 using oleic acid as an auxiliary material (polysaccharide-oleic acid-Rh2=4:1:2, drug loading 28.57%)

[0122] Accurately weigh 10 mg of Rh2 and 5 mg of oleic acid and ultrasonically dissolve them in 600 uL of anhydrous ethanol. Dissolve 20 mg of soybean polysaccharide in 5 mL of water. Add the ethanol solution dropwise to the polysaccharide solution under ultrasound. Remove the ethanol by rotary evaporation under reduced pressure at 45°C. Detect the particle size, PDI, and Zeta potential. The results are shown in Table 27 below. It can be seen that nanopolysaccharides were successfully prepared and ginsenoside Rh2 was successfully loaded.

[0123] After being placed at room temperature for one week, there was no significant change in particle size and PDI (Table 26), indicating that the drug-loaded nanopolysaccharide had good stability.

[0124] Table-27 Particle size, PDI, and potential of nanopolysaccharides loaded with human Rh2 using oleic acid as an excipient

[0125] Example 18 Preparation of Nano-lily Polysaccharide with Oleic Acid as an Excipient and Preparation of Composite Excipient-Compound Polysaccharide Nanoaggregates

[0126] Weigh 150 mg of lily polysaccharide (sugar content> 80%), astragalus polysaccharide, and lentinan, and prepare 5 mL of 50 mg / mL polysaccharide aqueous solution with deionized water. Accurately measure some oleic acid (OA), caprylic acid capric acid medium chain triglyceride Labrafac TM Lipophile WL 1349, polycaprolactone PCL with a molecular weight of 1000, and polylactic acid PLA with a molecular weight of 1000 were dissolved in ethanol to prepare a solution with a concentration of 10 mg / mL. An appropriate amount of ethanol solution was dropped into the polysaccharide solution under 250W ultrasonic conditions. The ethanol was removed by vacuum evaporation at 45°C, and the particle size, PDI and Zeta potential were tested. The results are shown in Table 28. It can be seen that nano-polysaccharides can be successfully prepared from single polysaccharide and single lipid excipients, and nano-polysaccharides can also be successfully prepared from single polysaccharide to composite excipients, composite polysaccharide to single excipients, and composite polysaccharide to composite excipients.

[0127] Table-28 Particle size, PDI, and potential of lily polysaccharide and composite excipients-complex polysaccharide nanoassemblies

[0128] Example 19 Preparation of Nanocarob Polysaccharide and Mixed Polysaccharide Nanoparticles: Oleic Acid Induction

[0129] Methods: Carob bean polysaccharide (Canopyrum sibiricum polysaccharide, PDI <1.6, polysaccharide content >93%) of uniform molecular weight (referred to herein as carob bean polysaccharide, abbreviated as CSPS) was precisely weighed and prepared into a 10 mg / mL solution with deionized water. Oleic acid (OA) (7.5 mg, 10 mg, and 15 mg) was precisely measured and dissolved in 1 mL of methanol. The solution was then added dropwise to the carob bean polysaccharide solution under 250W ultrasound. The methanol was removed by rotary evaporation at 35°C under reduced pressure to obtain CSPS nanopolysaccharide (CSPS@OA NPS). The particle size, PDI, and zeta potential were measured. The results are shown in Table 28, indicating that all nanopolysaccharides were successfully prepared. The particle size and size distribution of the nanocarob bean polysaccharide CSPS@OANPS with a polysaccharide:oleic acid ratio of 15:1 are shown in Figure 1.

[0130] The same method was used to prepare carob polysaccharide instead of carob total polysaccharide (containing the effective polysaccharide fraction of CSPS, with a polysaccharide content of about 82%). The results are shown in Table 29. It can be seen that nano-polysaccharides were successfully prepared, and it seems that purer carob polysaccharide is easier to prepare.

[0131] The total polysaccharide of carob, astragalus polysaccharide and wolfberry polysaccharide were mixed in equal amounts, and nano polysaccharides were prepared according to the above method at a total polysaccharide to oleic acid ratio of 9:1 (mass ratio). The results are shown in Table 28, which shows that the mixed polysaccharide was also successfully used to prepare nano polysaccharides.

[0132] Table-29 Particle size, PDI, potential of nanoassemblies of carob polysaccharide and its composite polysaccharide with oleic acid as auxiliary material

[0133] Example 20 Morphology of Nanocarob Polysaccharide CSPS@OA NPS

[0134] Carob polysaccharide CSPS and carob nanopolysaccharide CSPS@OA NPS were diluted with pure water to a polysaccharide content of approximately 1 mg / mL. Approximately 10 μL of each was dropped onto a 300-mesh copper stencil, allowed to air dry, and then stained with 2% (w / v) uranyl acetate for 2 minutes. The morphologies of the carob polysaccharide CSPS and carob nanopolysaccharide CSPS@OANPS were then observed at an accelerating voltage of 120 kV. As shown in Figure 2, the carob nanopolysaccharide CSPS@OANPS exhibited a regular spherical shape under a microscope.

[0135] Example 21 Particle size stability of nanocarob polysaccharide CSPS@OA NPS at room temperature and in physiological medium

[0136] To investigate the room temperature stability and particle size stability of carob bean nanopolysaccharides in physiological media, freshly prepared CSPS@OANPS were stored at 4°C and sampled at specific time points (1, 3, 5, 7, 9, 11, 13, and 15 days) to assess particle size and PDI. The results, shown in Figure 3, show that the carob bean nanopolysaccharides exhibited no significant changes in particle size and PDI over the storage period, demonstrating excellent stability.

[0137] To investigate the particle size stability of carob bean nanopolysaccharides in physiological media, CSPS@OANPS were mixed with equal volumes of 1.8% NaCl, 2× PBS, 10% glucose, or four volumes of artificial gastric fluid, artificial intestinal fluid, and mouse plasma at 37°C. Samples were taken at various intervals to measure particle size and PDI. The results, shown in Figure 4, show that the carob bean nanopolysaccharides exhibited no significant changes in particle size or PDI across the six physiological media tested, demonstrating excellent stability and suitability for direct oral or intravenous administration.

[0138] Example 22 Freeze-drying and reconstitution of nanocarob polysaccharide CSPS@OA NPS

[0139] The freshly prepared CSPS@OANPS was pre-frozen at -20°C and then freeze-dried. 30 mg of freeze-dried powder was added to 3 mL of pure water and shaken, and it was found that it could dissolve rapidly. The particle size after redissolution was measured to be 142±3.2, and the PDI value was 0.261±0.11, which were almost the same as the particle size and PDI value before freeze-drying. This shows that the nanocarob polysaccharide CSPS@OANPS can be freeze-dried without any freeze-drying protectant, which is convenient for long-term storage. It can be restored to a nanopolysaccharide by adding water and shaking before use.

[0140] Example 23 FITC Fluorescence Labeling of Carob Polysaccharide CSPS

[0141] 100 mg of carob bean polysaccharide (CSPS) was dissolved in 5 mL of water, mixed with 2.5 mL of 2.6 M NaOH solution and 40 μL of ammonium hydroxide, stirred at 40°C for 2 hours, transferred to a dialysis bag with a molecular weight cutoff of 8000-14000, dialyzed against water at 25°C for 48 hours, and freeze-dried. The freeze-dried residue was dissolved in 5.0 mL of 30% ammonium hydroxide and aminized at 40°C for 90 minutes. The reaction mixture was dialyzed against water for 48 hours (molecular weight cutoff 8000-14000) and freeze-dried. The freeze-dried sample was dissolved in 5 mL of 0.5 M sodium bicarbonate solution, 20 mg of FITC was added, stirred at room temperature in the dark for 24 hours, and precipitated by adding anhydrous ethanol to 80% (v / v). The precipitate was centrifuged at 5000 rpm for 10 minutes, the precipitate was collected, redissolved in water, dialyzed against water in the dark for 24 hours, and freeze-dried to obtain the CSPS-FITC fluorescently labeled product.

[0142] Example 24 Near-infrared fluorescence labeling of carob polysaccharide CSPS

[0143] 100 mg of carob polysaccharide CSPS was dissolved in 5 mL of water, mixed with 2.5 mL of 2.6 M NaOH solution and 40 μL of ammonium hydroxide, stirred at 40°C for 2 hours, transferred to a dialysis bag with a molecular weight cutoff of 8000-14000, dialyzed against water at 25°C for 48 hours, and freeze-dried. The lyophilized residue was dissolved in 5.0 mL of 30% ammonium hydroxide and aminized at 40°C for 90 minutes. The reaction mixture was dialyzed against water for 48 hours (molecular weight cutoff 8000-14000) and freeze-dried. The lyophilized powder was dissolved in 5 mL of 0.5 M sodium bicarbonate solution, 4 mg of IR783 was added, stirred in the dark at room temperature for 24 hours, and precipitated by adding anhydrous ethanol to 80% (v / v). The mixture was centrifuged at 5000 rpm for 10 minutes, the precipitate was collected, redissolved in water, dialyzed against water in the dark for 24 hours, and freeze-dried to obtain the CSPS-IR783 fluorescently labeled product.

[0144] Example 25 Study on the anti-inflammatory activity of Astragalus polysaccharide APS and nano Astragalus polysaccharide APS@OA NPS in vitro

[0145] RAW264.7 cells in the logarithmic growth phase were seeded in 96-well plates (at a density of 1×10 5Cells were cultured for 1 day. APS solution and nano-APS were diluted with fresh DMEM to different concentrations (0.1, 0.5, 1.0, 2.0, 5.0, 10, 20, 50, and 100 μg / mL CSPS). 200 μL was added to designated wells and incubated for 48 hours. Then, 10 μL of CCK-8 reagent was added to each well and incubated for 2 hours. Absorbance was measured at 450 nm using a microplate reader. Cell viability (%) was determined as follows: Cell inhibition rate (%) = (1 - OD1 / OD2) × 100%, where OD1 represents sample group and OD2 represents blank group. The results of the CCK-8 experiment showed that within 24 hours, free polysaccharides and nanopolysaccharides had no effect on the proliferation ability of RAW 264.7 cells in the range of 0-300 μg / mL (Figures 5A and 5B). Therefore, free polysaccharides and nanopolysaccharides at concentrations of 10, 100, and 300 μg / mL were selected to evaluate their in vitro anti-inflammatory effects on RAW 264.7 cells.

[0146] The macrophage RAW 264.7 inflammation model is widely used to evaluate the anti-inflammatory activity of drugs. RAW264.7 cells in logarithmic growth phase were seeded into 96-well plates at a density of 1×10 5 The cells were cultured for 24 hours and stimulated with 1 μg / mL LPS for 24 hours to establish a cellular inflammation model. The supernatant was discarded, and blank DMEM medium, polysaccharide solution (10, 100, and 300 μg / mL, diluted in fresh DMEM), and nanopolysaccharide (10, 100, and 300 μg / mL, diluted in fresh DMEM) were added. After incubation for 48 hours, 50 μL of Griess solution was inoculated into each well and incubated for 15 minutes. NO levels in the cells were measured using a commercial detection kit using a microplate reader according to the protocol provided. Similarly, the concentrations of tumor necrosis factor-α (TNF-α) and interleukins 12 and 6 (IL-12 and IL-6) in the cells were assessed using an enzyme-linked immunosorbent assay (ELISA) kit according to the protocol provided.

[0147] NO, IL-6, and TNF-α are important inflammatory indicators, and IL-10 is an important anti-inflammatory indicator. ELISA kits were used to detect the production of pro-inflammatory factors in RAW 264.7 cells after 24 hours of incubation with free polysaccharides and nano-polysaccharides. As can be seen from Figures 5C-5F, free polysaccharides can significantly reduce the secretion of NO, IL-6, and TNF-α, and significantly increase the production of IL-10, and in a dose-dependent manner, with good anti-inflammatory activity. At the same concentration (100 μg / mL or 300 μg / mL), nano-polysaccharides can more significantly reduce the levels of NO (P<0.05), IL-6 (P<0.01), and TNF-α (P<0.01) than free polysaccharides, and more significantly increase the level of IL-10 (P<0.05), indicating that nano-assembly greatly enhances the anti-inflammatory activity of polysaccharides.

[0148] Example 26 Study on the Antitussive, Expectorant and Anti-inflammatory Activities of Lily Polysaccharides and Nano-Lily Polysaccharides in Vivo

[0149] 90 male ICR mice weighing 21±2g were used. Inflammation and coughing were induced in mice by endotracheal intubation of lipopolysaccharide (LPS) combined with ammonia inhalation. The mice were fed adaptively for 3 days and then randomly divided into 9 groups (10 mice per group). The dosing schedule is shown in Table 30. The nano polysaccharide used in this study was the nano lily polysaccharide with oleic acid as an adjuvant in Example 18. Physiological saline was used as the dispersion medium for the intravenous polysaccharide solution, and the intravenous nano polysaccharide was adjusted to isotonicity with sodium chloride before administration.

[0150] Table-30 Grouping and Dosage Scheme

[0151] With the exception of the blank control group, LPS (0.4 mg / mL) was administered via intubation (10 μL / 10 g mouse body weight) on days 1, 7, and 15. On days 7 and 14, 0.2 mL of 25% ammonia solution was dripped onto a cotton ball and placed in a 1 L inverted beaker. Each mouse, except the blank control group, was quickly placed in the beaker for 2 minutes. Model establishment was considered successful if the model group mice exhibited frequent neck extension, mouth opening, back arching, abdominal muscle twitching, and coughing 20 or more times within 2 minutes.

[0152] Phenylpropyl phosphate tablets (60 mg / kg) were used as a positive drug to treat cough. One hour after the last administration, each mouse was placed in a 500 mL glass jar saturated with 0.2 mL of 25% ammonia water, and the latency and frequency of coughing were observed and recorded within 2 minutes.

[0153] Subsequently, the same mice were immediately evaluated for expectorant efficacy using the phenol red secretion assay, using ammonium chloride as the positive agent. Thirty minutes after intraperitoneal injection of XX μL of 3.5% phenol red solution, mice were anesthetized with 1% sodium pentobarbital (pentobarbital sodium) and sacrificed. The trachea and a segment of bronchus were immediately dissected and immersed in 1 mL of 5% sodium bicarbonate solution. The cells were sonicated for 15 minutes and centrifuged at 3000 rpm for 10 minutes. The supernatant was measured for absorbance at 546 nm using a microplate reader to calculate the phenol red content. A phenol red standard curve was constructed based on the literature.

[0154] Finally, anti-inflammatory activity was assessed by inducing acute airway inflammation in mice using ammonia. After sacrifice, the right lungs were harvested, washed with pre-chilled PBS, and homogenized using a high-throughput tissue grinder (lung tissue: PBS = 1 g: 3 mL). The supernatant was centrifuged at 1000 rpm for 10 minutes at 4°C, and the supernatant was collected for ELISA analysis of IL-6, IL-12, and TNF-a. Simultaneously, the left lungs were fixed by immersion in 4% formaldehyde for 24 hours, dehydrated with ethanol, embedded in paraffin, and sectioned at 4 μm for pathological observation using hematoxylin and eosin (H&E) staining.

[0155] Results: The antitussive effects of free polysaccharide and nanopolysaccharide were compared in vivo using an ammonia-induced cough model. The results are shown in Figures 6A and 6B. After 14 days of treatment, compared with the model control group, the number of coughs after oral administration of free polysaccharide decreased by 67.87% and the number of coughs after injection decreased by 88.97%. The number of coughs after oral administration of nanopolysaccharide decreased by 78.82% and the number of coughs after injection decreased by 97.21%, both significantly superior to the polysaccharide solution (P < 0.05) and the positive drug phenylpropyl phosphate (P < 0.05 and P < 0.01). Furthermore, the cough latency was significantly prolonged (Figure 6B), with the free polysaccharide increasing by 373.57% and 549.32% after oral and intravenous administration, respectively. The nanopolysaccharide increased the cough latency even more significantly, by 1.44 times (oral) and 1.54 times (intravenous) compared to the free polysaccharide (both P < 0.01), and significantly superior to the positive drug phenylpropyl phosphate (P < 0.05 and P < 0.01). Whether administered orally or intravenously, nanopolysaccharides were significantly more effective than polysaccharide solutions in treating cough (P < 0.05, P < 0.01). Intravenous administration was significantly more effective than oral administration in terms of both cough latency (polysaccharide P < 0.05, nanopolysaccharide P < 0.01) and the number of coughs within 2 minutes (P < 0.01 for both groups). Because large molecular weight polysaccharides are rarely absorbed into the circulation intact, this suggests that polysaccharides may have multiple targets in the body beyond the intestine, or that polysaccharides may have multiple pharmacologically active domains, and that degradation fragments containing these active domains can be absorbed into the circulation to exert their pharmacological activity.

[0156] The expectorant activity of free polysaccharides and nano-polysaccharides was detected by the secretion of phenol red in mice, and the results are shown in Figure 6C. Compared with the model control group, free polysaccharides significantly increased the secretion of phenol red in rats by 32.08% (oral administration) and 57.08% (intravenous administration), respectively. Nano-polysaccharides further promoted the secretion of phenol red, and the secretion of phenol red was 1.87 times that of oral administration of polysaccharide solution and 1.62 times that of intravenous injection. The results showed that at a dose of 100 mg / kg, free polysaccharides had a strong expectorant effect in vivo, and the expectorant effect of nano-polysaccharides was significantly better than that of free polysaccharides (P<0.05, P<0.01).

[0157] IL-10, TNF-α, and IL-6 are important pro-inflammatory chemokines and cytokines that play a key role in adaptive and innate immunity. Due to their pro-inflammatory properties, IL-10, TNF-α, and IL-6 are directly involved in the recruitment and activation of inflammatory cells at the site of injury. As shown in Figures 6D, 6E, and 6F, compared with the model group, the IL-10 level in the lung tissue of rats in the other groups was significantly increased (P<0.01, 0.001), the TNF-α level was significantly decreased (P<0.0001), and the IL-6 level was significantly decreased (P<0.01, 0.0001). The in vivo anti-inflammatory ability was as follows: oral administration of free polysaccharide < intravenous injection of free polysaccharide ≈ oral administration of nanopolysaccharide < intravenous injection of nanopolysaccharide (iv), among which intravenous injection of nanopolysaccharide had the strongest anti-inflammatory ability in vivo. Whether orally or intravenously administered, the in vivo anti-inflammatory activity of nanopolysaccharide was significantly higher than that of free polysaccharide (P<0.05, P<0.01).

[0158] H&E staining showed pathological changes in the lung tissue of mice (Figure 6G). The alveolar structure of the lung tissue of the blank group was clear, without inflammatory cell infiltration, and the alveoli were not fused. In the lung tissue of the model group, the alveolar wall was significantly thickened in the visual field, the alveolar mucosa was dysfunctional, the alveolar structure was disordered, and a small amount of alveolar fusion was observed. Lymphocytes and neutrophils were diffusely infiltrated, and there was a small amount of local bleeding. The alveolar wall in the oral free polysaccharide group was more thickened, accompanied by a small amount of lymphocyte and neutrophil infiltration. A small amount of alveolar wall thickening was observed in the free polysaccharide injection group, accompanied by a small amount of lymphocyte, neutrophil and macrophage infiltration. The alveolar structure of the nanopolysaccharide oral group was clear. No alveolar wall thickening was observed in the nanopolysaccharide injection group, the inflammatory cell infiltration of the tube wall was reduced, the bronchial smooth muscle was thinned, and the alveolar structure was intact. It can be seen that nanopolysaccharides can better repair damage caused by lung inflammation than free polysaccharides.

[0159] Example 27 In vitro cellular uptake study of carob polysaccharide CSPS and nanocarob polysaccharide CSPS@OANPS

[0160] Caco-2 cells in the logarithmic growth phase were seeded into 24-well plates (1×10 6Cells were plated at 400 nmol / well and incubated at 37°C and 5% CO2 for 24 hours. FITC-labeled free polysaccharide (FITC-CSPS) and nanopolysaccharide prepared using FITC-CSPS (FITC-CSPS@OANPS) were diluted to 100 μg / mL in serum-free culture medium, added to the wells, and mixed thoroughly. The cells were incubated in an incubator for 1, 3, and 6 hours, respectively. The culture medium was discarded, and DAPI (5 μg / mL, 0.4 mL) was added. The cells were fixed in 0.4 mL of PBS containing 4% (w / v) paraformaldehyde. Cellular uptake of FITC-CSPS and FITC-CSPS@OANPS was observed using an inverted fluorescence microscope. Semi-quantitative fluorescence analysis and colocalization analysis were performed using Image J software.

[0161] As shown in Figure 7, Caco-2 cells exhibited time-dependent cellular uptake of both free and nano-polysaccharides. However, at the same concentration of 100 μg / mL, cellular uptake of the nano-polysaccharide was significantly enhanced compared to the free polysaccharide, with up to 5-fold enhancement (P < 0.0001), particularly at the third and sixth hours. This portion of the experiment demonstrates a significant increase in cellular uptake of the nano-polysaccharide relative to the free polysaccharide. This finding partially explains why the nano-polysaccharide exhibits superior anti-inflammatory activity compared to the polysaccharide solution.

[0162] Example 28 Pharmacokinetic Study of Carob Polysaccharide CSPS and Nanocarob Polysaccharide CSPS@OANPS

[0163] Mouse blank plasma was mixed with near-infrared fluorescent-labeled carob polysaccharide solution (CSPS-IR783) to prepare a series of solutions with polysaccharide concentrations of 1, 2.5, 5, 10, 20, 40, 80, 160, 320, and 640 μg / mL to obtain a standard curve.

[0164] Sixty-six ICR mice were randomly divided into two groups and administered a near-infrared fluorescent-labeled carob polysaccharide solution (CSPS-IR783) or carob nanopolysaccharide (CSPS-IR783@OANPs) via the tail vein at a dose of 50 mg / kg. Blood samples were collected from three mice in each group at 0.5, 1, 2, 3, 4, 6, 8, 10, 12, 24, and 48 hours after administration. Heparinized blood samples were centrifuged at 3500 rpm for 10 minutes at 4°C. Plasma samples were collected and mixed with saline (1:4, volume ratio) and centrifuged at 3500 rpm for 10 minutes. The supernatant was collected and fluorescence intensity was measured using a microplate reader (excitation wavelength 745 nm, emission wavelength 820 nm). Polysaccharide concentrations were calculated from the standard curve. Plasma polysaccharide concentrations were plotted over time to calculate pharmacokinetic parameters.

[0165] Results: As shown in Figure 8, after a single intravenous injection of polysaccharide solution, free polysaccharide was quickly cleared from the circulation, while the clearance rate of nano-polysaccharide was slightly slower. The pharmacokinetic parameters calculated by DAS2.0 software are shown in Table 31. The metabolism of free polysaccharide and nano-polysaccharide in mice was close to the two-compartment model, and the fitting constant R 2 Compared with free polysaccharide, although Cmax MRT(0→∞) and Vd were only slightly increased, nanopolysaccharide significantly increased AUC(0→t) (1.17-fold, P<0.0001) due to the significant prolongation of t(1 / 2)α and t(1 / 2)β. This result partially explains why nanopolysaccharide has better antitussive, expectorant, and anti-inflammatory effects in vivo than free polysaccharide when injected intravenously.

[0166] Table 31 Pharmacokinetic parameters of free polysaccharides and nanopolysaccharides labeled with IR783 after intravenous injection in mice

[0167] Example 29 Congo Red Analysis of Carob Polysaccharide CSPS and Nanocarob Polysaccharide CSPS@OANPS

[0168] Congo red is a powdered acidic dye that is readily soluble in ethanol and water. It is primarily used to detect the presence of triple-helical structures in polysaccharides. The principle behind this is its ability to form complexes with polysaccharides with multiple triple-helical structures. Violet light spectroscopy reveals a relationship between the maximum absorption wavelength and the concentration of sodium hydroxide (NaOH). Specifically, when the NaOH concentration exceeds 0.2 mol / L, the maximum absorption wavelength decreases significantly. Therefore, based on this change in the maximum absorption wavelength, we can determine whether a polysaccharide has a triple-helical structure.

[0169] Methods: 10 mg of carob bean polysaccharide nanoparticles (CSPS@OA NPs) were accurately weighed and dissolved in 5 ml of ultrapure water to prepare a 2 mg / ml solution. An appropriate amount of Congo red was weighed and dissolved in ultrapure water to prepare a 160 μg / ml Congo red solution. NaOH solutions with concentrations of 0.4, 0.8, 1.2, 1.6, and 2.0 mol / L were prepared and used. In 10 separate test tubes, carob bean polysaccharide nanoparticles (CSPS@OA NPs), Congo red solution, and NaOH solution were added to five of the tubes at a 2:1:1 ratio, resulting in a final polysaccharide concentration of 1 mg / ml. Ultrapure water, Congo red solution, and NaOH solution were also added to five of the tubes at a similar 2:1:1 ratio. The final concentration of Congo red in each test tube was 40 μg / ml, and the final concentrations of NaOH were 0.1, 0.2, 0.3, 0.4, and 0.5 mol / L, respectively. The samples were thoroughly shaken and mixed, and allowed to stand at room temperature for 10 min. Ultrapure water was used as a blank control, and a UV spectrophotometer was used to scan at a wavelength of 400-600 nm, and the maximum absorption wavelength at different NaOH concentrations was recorded.

[0170] A plot was drawn with NaOH concentration (mol / L) and maximum absorption wavelength (nm) as the horizontal and vertical axes, respectively. The results, shown in Figure 9, show that the maximum absorption wavelength of the complex formed by carob polysaccharide and Congo red increases slightly in the range of 0-0.2 mol / L of sodium hydroxide and decreases sharply in the range of 0.2-0.3 mol / L, demonstrating that carob polysaccharide has a typical triple helical structure. After carob polysaccharide was prepared into nanopolysaccharide, the maximum absorption wavelength of the complex formed with Congo red matched that of free carob polysaccharide, indicating that the process of preparing the nanopolysaccharide did not affect the triple helical higher-order structure of carob polysaccharide in any way.

[0171] Example 30 Comparison of Fourier infrared spectra of carob polysaccharide CSPS and nanocarob polysaccharide CSPS@OANPS

[0172] 5.0 mg of CSPS and CSPS@OANPs samples were accurately weighed, added with an appropriate amount of dry KBr, and ground into uniform transparent pellets. The pellets were then analyzed by Fourier transform infrared spectrometer within the range of 4000 cm-1 to 400 cm-1. The results are shown in FIG10 .

[0173] Fourier transform infrared spectroscopy (FT-IR) is a method for identifying molecular structure or quantifying it, and is also a common technique for analyzing the fine structure of macromolecules such as polysaccharides. As shown in Figure 10, the FT-IR images of the nanopolysaccharide and the free polysaccharide have excellent overlap, demonstrating that the nanopolysaccharide preparation process does not affect the fine structure of the polysaccharide.

[0174] Example 31. Preparation of Nanopolysaccharides by Direct Dispersion Method Using Amphiphilic Lipid Materials as Excipients

[0175] Total carob polysaccharides were precisely weighed and dissolved in water to prepare a 10 mg / mL polysaccharide solution. 6 mg each of TPGS, PLA-1000-PEG2000, PCL1000-PEG2000, and PLGA1000-PEG2000 were weighed and added to 5 mL of the polysaccharide solution. The solution was magnetically stirred for 5 minutes and sonicated in a 250W waterbath for 10 minutes. The particle size and PDI (Physical Indicator) were measured (see Table 32). The solution was then homogenized ten times at 1400 bar, and the particle size and PDI were measured again (see Table 32). This indicates that lipid materials containing PEG segments can also be dispersed in the polysaccharide solution, stirred, and sonicated to obtain polysaccharide nanoassemblies. High-pressure homogenization can further reduce the particle size and size distribution of the resulting nanopolysaccharide.

[0176] Table-32 Particle size, PDI, and potential of nanopolysaccharides prepared by direct dispersion method using amphiphilic lipid materials as excipients

[0177] Example 32. Nanopolysaccharide loaded with macromolecular drugs

[0178] Accurately weigh carob total polysaccharides and dissolve them in water to prepare a 10 mg / mL polysaccharide solution. 5 mg of insulin, 5 mg of bevacizumab, and 5 mg of rituximab were accurately taken to prepare 1 mL of solution, which was added to 4 mL of the polysaccharide solution and mixed thoroughly to obtain three mixed solutions, each 5 mL. Separately, 3 mg of oleic acid and 3 mg of TGPS were dissolved in 0.5 mL of ethanol. The ethanol solution was slowly added dropwise to the mixed solution under gentle stirring at 100 rpm. The ethanol was evaporated under reduced pressure at 35°C, and the particle size and particle size distribution were measured. The results are shown in Table 33. It can be seen that nanopolysaccharides can be successfully loaded with molecular drugs such as peptides and proteins.

[0179] After being placed in a refrigerator at 35°C for 3 days, the particle size and particle size distribution of the nanopolysaccharide loaded with macromolecular drugs were re-measured. The results are shown in Table 32. It can be seen that the obtained nanopolysaccharide loaded with macromolecular drugs has good storage stability.

[0180] Table-33 Drug-loaded nanopolysaccharides obtained by loading peptides and proteins with nanopolysaccharides and their particle size and PDI after placement

[0181] The above description of the disclosed embodiments is intended to enable one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be readily apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention is not limited to the embodiments shown herein but is intended to conform to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A polysaccharide composition, characterized in that The polysaccharide composition comprises polysaccharides and lipid materials; wherein, The polysaccharides include animal polysaccharides, plant polysaccharides, marine polysaccharides, fungal polysaccharides and bacterial polysaccharides; The lipid material is selected from the group consisting of medium-chain and long-chain fatty alcohols (C6-C50) and their esters and ethers, medium-chain and long-chain fatty amines (C6-C50) and their amides formed with acids, medium-chain and long-chain fatty acids (C6-C50) and their esters and derivatives, amides of medium-chain and long-chain fatty acids (C6-C50) and their derivatives, amphiphilic materials containing medium-chain and long-chain fatty chains (C6-C50), molecular weight Long-chain fat-soluble materials with a molecular weight less than 10,000 and amphiphilic polymers with them as lipophilic segments, terpenoid compounds with side fatty chains and their glycosides, other small molecules with fatty chains and amphiphilic molecules with them as lipophilic parts, and the fatty chains of the lipid materials are saturated fatty chains or contain one, two, or more unsaturated bonds; The weight ratio of the polysaccharide to the lipid material is 2-50:

1.

2. The polysaccharide composition according to claim 1, characterized in that The lipid material is selected from medium-chain and long-chain fatty alcohols (C8-C30) and their esters and ethers, medium-chain and long-chain fatty amines (C8-C30) and amides formed with acids, medium-chain and long-chain fatty acids (C6-30) and their esters and derivatives, amides of medium-chain and long-chain fatty acids (C8-C30) and their derivatives, amphiphilic materials containing medium-chain and long-chain fatty chains (C8-C30), long-chain fat-soluble materials with a molecular weight of <5000, and amphiphilic polymers with them as lipophilic segments.

3. The polysaccharide composition according to claim 2, characterized in that The lipid material is selected from medium-chain and long-chain fatty alcohols (C8-C18) and their esters and ethers, medium-chain and long-chain fatty amines (C8-C18) and amides formed with acids, medium-chain and long-chain fatty acids (C8-C18) and their esters and derivatives, amides of medium-chain and long-chain fatty acids (C8-C18) and their derivatives, amphiphilic materials containing medium-chain and long-chain fatty chains (C8-C18), long-chain fat-soluble materials with a molecular weight of <3000, and amphiphilic polymers with them as lipophilic segments.

4. The polysaccharide composition according to claim 1, characterized in that The polysaccharide composition further comprises pharmaceutically acceptable excipients, which include common additives for injections, adhesives, disintegrants, lubricants, tablet fillers / diluents, and tablet lubricants.

5. A nano polysaccharide, characterized in that The nano polysaccharide is prepared from the polysaccharide composition according to claim 1 by dissolving a lipid material in a water-miscible organic solvent, adding the solution to the polysaccharide under stirring or ultrasound to induce self-assembly of the polysaccharide, and then removing the organic solvent to allow the lipid material and the polysaccharide to assemble into nanoparticles with a particle size of 20-1000 nm. Alternatively, the lipid material is an amphiphilic lipid material, and the nanoparticles are obtained by dispersing the amphiphilic lipid material in an aqueous solution of polysaccharide, stirring, and ultrasonicating.

6. A method for preparing nano polysaccharide according to claim 5, characterized in that: The method specifically comprises the following steps: 1) dissolving the lipid material in ethanol, methanol, acetone, isopropanol or a mixed solvent thereof to obtain a lipid material solution, and dissolving the polysaccharide in water to obtain a polysaccharide solution; Alternatively, the method may be as follows: dissolving the polysaccharide in water to obtain a polysaccharide solution; 2) adding the lipid material solution to the polysaccharide solution under stirring or ultrasound, and evaporating the solvent to obtain nano-polysaccharides with an average particle size of 20-1000 nm; Alternatively, the amphiphilic lipid material is dispersed in an aqueous solution of polysaccharide, and the mixture is stirred and ultrasonicated to obtain nano-polysaccharide; 3) If the particle size of the obtained nanopolysaccharide is too large or the particle size distribution is too wide, the particle size and / or particle size distribution can be further reduced by high-pressure homogenization; 4) The obtained nano-polysaccharide can be further spray-dried or freeze-dried to become a solid powder to suit different uses.

7. The method for preparing nano polysaccharide according to claim 6, characterized in that: The evaporation in step 2) is one of natural evaporation, heating evaporation, and reduced pressure evaporation, or a combination thereof. The homogenization pressure in step 3) is 500-4000 bar.

8. Use of the polysaccharide composition according to claim 1 or the nano polysaccharide according to claim 5 in the preparation of food, health products, and pharmaceutical products.

9. The use according to claim 8, characterized in that The nano polysaccharide can be further loaded with fat-soluble small molecules or protein, polypeptide, nucleic acid macromolecules to become nano particles with health care and therapeutic effects.

10. The use according to claim 8, characterized in that The nano-polysaccharide solution and its dry powder can also be further processed as intermediates into various forms of food, health products or pharmaceutical products, including oral liquid, suspension, gel, jelly, granules, tablets, capsules, inhalation preparations, spray preparations, injections, freeze-dried powder for injection, ointments, patches, microneedles, for oral administration, injection, mucosal, cavity, wound application or external use.

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