Delivery carrier

Abstract

The invention discloses a stevioside derivative, and the derivative or an in-vivo metabolite thereof has excellent hydrophilic-lipophilic balance and can be used as a delivery carrier of an active component. The invention also discloses a preparation method and application of the derivative.

Description

Delivery vehicle Technical Field

[0001] The present invention relates to the field of medical technology, and specifically to a steviol glycoside derivative that can be used as a drug carrier, a preparation method and use thereof. Background Art

[0002] Steviosides are natural sweeteners extracted from the herb Stevia rebaudiana, a member of the Asteraceae family. Stevia has been used as a medicinal herb and as a sugar substitute in South America for centuries. Currently, over 100 compounds have been isolated and identified from stevia. Among them, stevioside, rebaudioside A, rebaudioside C, and dulcoside A are found in high concentrations in stevia leaves and can be directly extracted and purified.

[0003] Currently, stevioside, rebaudioside-A, B, C, D, E, M, and AM are listed as GRAS materials as food additives, and the acceptable daily dose for humans is 0-4 mg / kg body weight. The general structural formula of these compounds is shown in (1):

[0004] G is selected from the following groups: The two Gs in the general formula (1) may be the same or different,

[0005] Steviosides are structurally classified as kaurene-type tetracyclic diterpenes. Their chemical structure has a hydrophobic core of steviol and highly hydrophilic sugar residues or hydroxyl groups at both ends. This structure is the main reason for the poor permeability of steviol glycosides as delivery vehicles. Nanoscale 2011, 3, 4564, Langmuir 2015, 31, 13664, Angew. Chem. Int. Ed. 2015, 54, 5408, and Fitoterapia 151 (2021) 104845 reported that natural triterpenes such as ursolic acid, betulinic acid, oleanolic acid, glycyrrhizic acid, and isoglycyrrhizic acid can self-assemble under certain conditions. European Journal of Drug Metabolism and Pharmacokinetics (2022) 47:827-839 reported that patients with type 2 diabetes took 3 grams of rebaudioside A orally. Two hours after oral administration, the peak concentration Cmax of rebaudioside A in plasma was approximately 178 ng / ml, indicating that the permeability of rebaudioside A itself is very poor.

[0006] Therefore, existing steviol glycoside compounds, such as stevioside, rebaudioside A, B, C, D, E, M, and AM, are not excellent delivery carriers. Except for rebaudioside B, other steviol glycosides are neutral under physiological conditions and are prone to aggregation as nanocarriers. They have poor chemical structure compatibility with charged carriers commonly used in lipid nanodelivery systems and are difficult to form stable assemblies.

[0007] Currently, there are no reports on structural modification and performance optimization of steviol glycosides for use as delivery vehicles. This study addresses the challenges of using natural steviol glycosides as delivery vehicles by designing and synthesizing a series of derivatives. Through extensive in vitro and in vivo studies, we identified steviol glycoside derivatives with superior delivery properties.

[0008] Summary of the Invention

[0009] The present invention aims, on the one hand, to improve the lipid solubility of steviol glycoside compounds as delivery vehicles or their metabolic intermediates in vivo by structurally modifying them, while ensuring that the modified compounds have a certain solubility. Another aspect of the present invention is to design new compounds that, when used as delivery vehicles, possess dissociable groups under physiological conditions (pH 1.0-pH 8.0), providing a certain charge to the assembled nanoparticles, thereby improving the stability of the nanoparticles and, more importantly, facilitating their passage through the negatively charged phospholipid bilayer of cell membranes.

[0010] Explanation of terms:

[0011] The term "carrier material" or "adjuvant" or "functional carrier material" refers to a material that can assist or carry one or more other substances into the body. This material may affect the odor, solubility, stability, mucosal irritation, hydration rate, release, absorption, tissue distribution, metabolism or clearance of the delivered substance during the delivery process or in the body.

[0012] The term "enantiomeric excess" or "ee value" refers to a measure of how much of one enantiomer is present compared to the other enantiomer. For a mixture of R and S enantiomers, the enantiomeric excess is calculated as ee value = ([R] - [S] / [R] + [S]) × 100%, where R and S are the respective molar or mass fractions of the enantiomers in the mixture.

[0013] The term "diastereomeric excess" or "de value" refers to a measure of how much of one diastereomer is present compared to the other diastereomer and is defined similarly to enantiomeric excess. Thus, for a mixture of diastereomers D1 and D2, the diastereomeric excess is calculated as de value = ([D1] - [D2] / [D1] + [D2]) x 100%, where D1 and D2 are the respective molar or mass fractions of the diastereomers in the mixture.

[0014] It is possible to determine diastereomeric excess and / or enantiomeric excess using analytical techniques including conventional nuclear magnetic resonance spectroscopy, high performance liquid chromatography based on chiral analytical columns, supercritical fluid chromatography, optical polarimetry, and the like, as will be understood by those skilled in the art.

[0015] The term "hyperbranched" refers to a class of highly branched three-dimensional molecules. This branched structure gives them rich terminal functional groups, making them easy to modify and more conducive to forming self-assembled complexes with other substances through non-covalent bonds. These molecules can be small molecules or high molecular polymers.

[0016] The technical solutions of the present invention are as follows:

[0017] 1. A delivery vehicle, characterized in that the delivery vehicle is a compound represented by general formula Ia, or a salt thereof, or a hydrate thereof, or any mixture of a compound represented by general formula Ia, a salt thereof, and a hydrate thereof,

[0018] In formula Ia, R1 and R2 are independent of each other, R1 is selected from hydroxyl or saccharide, and R2 is selected from

[0019] Wherein R3 is selected from alkali metals, alkaline earth metals, H, C1-C10 straight chain, branched, cyclic alkanes or alkanes whose side chains are substituted with hydroxyl, amino, or thiol, polymer groups or sugar groups; R4 and R5 are independently selected from H, C1-C10 straight chain, branched, cyclic alkanes or alkanes whose side chains are substituted with hydroxyl, amino, or thiol, polymer groups, amino acids or amino acid residues, aminoamides or short peptides condensed with any number of 1 to 9 amino acids.

[0020] 2. The delivery vector according to item 1, characterized in that the sugar group is a pyranose monosaccharide or oligosaccharide.

[0021] 3. The delivery vector according to item 1, characterized in that the sugar group is selected from pyranose glucose, galactopyranose, pyranose mannosyl, pyranose deoxyglucose, pyranose rhamnose, pyranose xylose, pyranose glucosamine, and oligosaccharides formed by polymerization of any number of 1 to 9 pyranose monosaccharides at any position.

[0022] 4. The delivery vector according to any one of items 1 to 3, characterized in that the absolute configuration of the sugar group is D-type or L-type, and the relative configuration of the terminal carbon is α-type or β-type.

[0023] 5. The delivery vehicle according to item 1, wherein the alkali metal is selected from sodium, potassium or lithium.

[0024] 6. The delivery vehicle according to item 1, wherein the polymer group is selected from polyethylene glycol, polydopamine, polyarginine amide or polyglutamic acid.

[0025] 7. The delivery vector according to item 1, characterized in that the amino acid or amino acid residue is selected from alanine, arginine, aspartic acid, cysteine, glutamic acid, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, tyrosine or valine, and the aminoamide is selected from the aminoamide corresponding to the amino acid.

[0026] 8. The delivery vector according to item 1, characterized in that the short peptide is a cell-penetrating-peptide (CPP).

[0027] 9. The delivery vector according to item 8, characterized in that the cell-penetrating peptide is octaarginine.

[0028] 10. The delivery vector according to item 1, characterized in that the R1 is selected from hydroxyl,

[0029] 11. The delivery vector according to item 1, characterized in that the R2 is selected from wherein n is 0-9, and X=OH and / or NH2.

[0030] 12. The delivery vector according to item 1, characterized in that the optical purity of the compound represented by general formula Ia is not limited, and it has R and / or S configuration, and has any enantiomeric excess value or any diastereomeric excess value.

[0031] 13. A delivery carrier, which is at least one compound selected from the group consisting of:

[0032] (1) (1R,4S,5R,9S,10R,13S)-13-[(2S,3R,4S,5R,6R)-5-hydroxy-6-(hydroxymethyl)-3,4-bis[[(2S,3R),4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl]oxy]oxa-2-yl]oxy-5,9-dimethyl-14-methylenetetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylate sodium (RBDS-B-Na),

[0033] (2) (4R, 4aS, 6aR, 9S, 11aR, 11bS)-N-((S)-1-amino-3-(1H-imidazol-4-yl)-1-oxopropan-2-yl)-9-((2S, 3R, 4S, 5R, 6R)-5-hydroxy-6-(hydroxymethyl)-3,4-bis(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide (RBDS-B-His),

[0034] (3) (4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-((2S, 3R, 4S, 5R, 6R)-5-hydroxy-6-(hydroxymethyl)-3,4-bis((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-N-((S)-2-hydroxypropyl)-4,11-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide (RBDS-B-APL),

[0035] (4) (1R,4S,5R,9S,10R,13S)-13-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl]oxyoxa-2-yl]oxy-5,9-dimethyl-14-methylenetetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylic acid (STVB),

[0036] (5) (6S, 9S, 12S, 15S, 18S, 21S, 24S, 27S)-1-amino-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-(((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy ... Hydrogen-1H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetradecahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-9,12,15,18,21,24,27-hepta(3-guanidinopropyl)-1-imino-7,10,16,19,22,5-heptyloxy-2,8,11,14,17,20,23,26-octaazaoctadecane-28 oleic acid (STVB-CPP-Arg8),

[0037] (6)(4R, 4aS, 6aR, 9S, 11aR, 11bS)-N-((S)-1-amino-3-(1H-imidazol-4-yl)-1-oxopropan-2-yl)-9-((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide (STVB-His),

[0038] (7) (4R, 4aS, 6aR, 9S, 11aR, 11bS)-N-((S)-1-amino-5-((diaminoethylene)amino)-1-oxopentan-2-yl)-9-((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide (STVB-Arga),

[0039] (8) (4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-N-((S)-2-hydroxypropyl)-4,11-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide (STVB-APL),

[0040] (9)(4R, 4aS, 6aR, 9S, 11aR, 11bS)-N-((S)-1-amino-5-((diaminoethylene)amino)-1-oxopentan-2-yl)-4,11-dimethyl-8-methylene-9-((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetradecahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide (RBSD-Arga),

[0041] (10) (6S, 9S, 12S, 15S, 18S, 21S, 24S, 27S)-1-amino-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-4,11b-dimethyl-8-methylene-9-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-1-amino-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-4,11b-dimethyl-8-methylene-9-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl) )oxy)tetradecahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-9,12,15,18,21,24,27-heptadeca(3-guanidinopropyl)-1-imino-7,10,13,16,19,22,25-heptyloxy-2,8,11,14,17,20,23,26-octaazaoctadecane-28-carboxylic acid (RBSD-CPP-Arg8),

[0042] (11) (6S, 9S, 12S, 15S, 18S, 21S, 24S, 27S)-1-amino-9,12,15,18,21,24,27-heptyl(3-guanidinopropyl)-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-hydroxy-4,11-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-amide)-1-imino-7,10,13,16,19,22,25-heptyloxy-2,8,11,14,17,20,23,26-octaazaoctacosanoic acid (STL-CPP-Arg8),

[0043] (12) (6S, 9S, 12S, 15S) -1-amino-9, 12, 15-tris (3-guanidinopropyl) -6- ((4R, 4aS, 6aR, 9S, 11aR, 11bS) -9-hydroxy-4, 11-dimethyl-8-methylenetetrahydro-6a, 9-methylcyclohepta[a]naphthalene-4-carboxamide) -1-imino-7, 10, 13-trioxy-2, 8, 11, 14-tetraazahexadecane-16-carboxylic acid (STL-CPP-Arg4),

[0044] (13) (S)-5-guanidine-2-((S)-5-guanidine-2-(4R,4aS,6aR,9S,11aR,11bS)-9-hydroxy-4,11-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-amido)pentanamido)pentanoic acid (STL-CPP-Arg2),

[0045] (14)(6S, 9S, 12S, 15S, 18S, 21S, 24S)-1-amino-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-(((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl) oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-9,12,15,18,21,24-hexa(3-guanidinopropyl)-1-imino-7,10,13,16,19,22-hexaoxo-2,8,11,14,17,20,23-heptaazapentacan-25 acid (STVB-CPP-Arg7),

[0046] (15)(6S, 9S, 12S, 15S, 18S, 21S)-1-amino-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-(((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)-1-yl )oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-9,12,15,18,21-penta(3-guanidinepropyl)-1-imino-7,10,13,16,19-pentaoxo-2,8,11,14,17,20-hexaazadocosa-22-oic acid (STVB-CPP-Arg6),

[0047] (16) (6S, 9S, 12S, 15S, 18S)-1-amino-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-(((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran- 2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-9,12,15,18-tetrakis(3-guanidinopropyl)-1-imino-7,10,13,16-tetraoxo-2,8,11,14,17-pentaazanonadecan-19-oic acid (STVB-CPP-Arg5),

[0048] (17) (6S,9S,12S,15S)-1-amino-6-((4R,4aS,6aR,9S,11aR,11bS)-9-(((2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H- (pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-9,12,15-tris(3-guanidinepropyl)-1-imino-7,10,13-trioxy-2,8,11,14-tetraazahexadecane-16-oic acid (STVB-CPP-Arg4),

[0049] (18) (S)-2-((S)-2-(S)-2-[(4R,4aS,6aR,9S,11aR,11bS)-9-((2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-1H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-5-guanidinecarboxamide)-5-guanidinecarboxamide)-5-guanidinecarboxamide]-5-guanidinecarboxylic acid (STVB-CPP-Arg3),

[0050] (19) (S)-2-((S)-2-((4R,4aS,6aR,9S,11aR,11bS)-9-((2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-1H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-5-guanidine amide)-5-guanidine valeric acid (STVB-CPP-Arg2), and

[0051] (20) (S)-2-((4R,4aS,6aR,9S,11aR,11bS)-9-((2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-1H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-5-guanidine pentanoic acid (STVB-CPP-Arg).

[0052] 14. A method for preparing the delivery vector of item 1, comprising the following steps:

[0053] (1) Step 1: Compound 1 is subjected to a hydrolysis reaction to obtain Compound 2, wherein when Compound 1 is in the form of a carboxylic acid, Step 2 is directly performed;

[0054] (2) Step 2: Protect the R1 group in compound 2 with a protecting group P to obtain compound 3;

[0055] (3) Step 3: subjecting compound 3 to a condensation reaction to obtain compound 4;

[0056] (4) Step 4: Compound 4 is subjected to a deprotection reaction to obtain a compound of formula Ia, wherein R1 in compound 1, compound 2, compound 3 and R2 in compound 4 are as defined in any one of items 1-12.

[0057] 15. The preparation method according to item 14, characterized in that the hydrolysis reaction is carried out in a solvent (preferably water) under alkaline conditions (preferably sodium hydroxide or potassium hydroxide), wherein compound 1 is selected from Stevioside (STVS), Rebaudioside A (RBDS-A), Dulcoside A (DCS-A), Rubusoside (RBSD), Rubusoside acid (RBSD-Acid), Steviolbioside (STVB), Rebaudioside B (RBDS-B), Rebaudioside C (RBDS-C) acid (RBDS-C-Acid), Steviol (STL) or Isosteviol; or characterized in that when compound 1 is Steviolbioside (STVB), Rebaudioside B (RBDS-B), Rebaudioside When the acid form of C (RBDS-C) (RBDS-C-Acid), steviol or rubusoside (RBSD) (RBSD-Acid), the preparation method does not require step 1. 16. The preparation method according to item 14 or 15, characterized in that the protecting group in step 2 is a protecting group of sugar or hydroxyl, preferably a silyl group or an acetyl group, preferably the sugar group or hydroxyl group is fully acetylated, for example, the protected compound 3 is selected from RBDS-B-Ac, STVB-Ac, RBSD-Acid-Ac, DCS-A-Acid-Ac or STL-Ac, and the structural formulas are shown below, respectively.

[0058] Wherein: Ac represents acetyl group.

[0059] 17. The preparation method according to any one of items 14 to 16, characterized in that the condensation reaction in step 3 is a reaction of compound 2 with a sugar, amine or alcohol having an R2 group to form a glycoside, amide or ester. Preferably, the carboxyl group of compound 2 is activated using oxalyl chloride to form an acyl chloride, which is then reacted with an amino compound in the presence of alkaline conditions (such as triethylamine).

[0060] 18. The preparation method according to any one of items 14 to 17, wherein when the protecting group P is an acetyl group, step 4 is a deprotection reaction performed using an alcohol solvent (such as methanol or ethanol) under strong alkaline conditions (such as potassium tert-butoxide, sodium methoxide, or sodium ethoxide).

[0061] 19. A delivery kit, characterized by comprising the delivery vehicle according to any one of items 1 to 13 and an active ingredient.

[0062] 20. The delivery kit according to item 19, wherein the active ingredient is a biologically active agent, a chemically active agent or an adjuvant.

[0063] 21. The delivery kit according to item 19 or 20, characterized in that the active ingredient is selected from: proteins / polypeptides, polysaccharides / oligosaccharides, nucleic acids or nucleic acid fragments, nanocarriers (such as liposomes, polymer micelles, inorganic nanoparticles), lipids, nutrients, organic small molecule compounds, phage particles, superparamagnetic substances, vaccines, cells, or any combination thereof.

[0064] 22. The delivery kit according to any one of items 19-21, characterized in that the weight ratio of the active ingredient to the delivery vehicle is about 1:0.5-1:150 (preferably 1:1, 1:10, 1:20, 1:30, 1:40, 1:50, 1:60, 1:70, 1:80, 1:90, 1:100, 1:110, 1:120, 1:130 or 1:140).

[0065] 23. The delivery kit according to any one of items 19 to 22, wherein one of the active ingredient and the delivery system is in a liquid state or a solid state, or the active ingredient and the delivery system are both in a liquid state or a solid state.

[0066] 24. The delivery kit according to any one of items 19 to 23, characterized in that the active ingredient and the delivery vehicle are present independently, or the active ingredient and the delivery vehicle are present in a mixture.

[0067] 25. The delivery kit according to any one of items 19 to 24, characterized in that the active ingredient and the delivery vehicle are in a form suitable for oral, sublingual, buccal, intraduodenal, intracolonic, rectal, vaginal, mucosal, subcutaneous, intramuscular, intravenous, arterial, cutaneous, pulmonary, intranasal, aural, or ocular administration.

[0068] 26. The delivery kit according to any one of items 19 to 25, characterized in that the active ingredient and / or the delivery vehicle is in the form of micelles, liposomes, nanoparticles, microspheres, microcapsules, solid dispersions, molecular compositions or hydrogels.

[0069] 27. The delivery kit according to any one of items 19 to 26, characterized in that when the active ingredient and the delivery vehicle exist independently, the two are assembled by mixing and dissolving them during use; when the active ingredient and the delivery vehicle exist in a mixed state, the active ingredient and the delivery vehicle in a solid state are assembled by dissolving them.

[0070] 28. The delivery kit according to any one of items 19 to 27, characterized in that the assembly of the active ingredient and the delivery vehicle in the delivery system is in situ assembly.

[0071] 29. The delivery kit according to item 28, characterized in that the in situ assembly is self-assembly in the gastrointestinal tract, local self-assembly in the mucosa, self-assembly in the blood, or self-assembly in target cells.

[0072] 30. The delivery kit according to any one of items 28-29, characterized in that the in situ assembly is the in situ assembly of the active ingredient with the metabolite of the delivery vector according to any one of items 1-13 after enzyme or microbial metabolism.

[0073] 31. The delivery kit according to any one of items 19 to 25, characterized in that the active ingredient and / or the delivery vehicle is in a dosage form selected from the following: tablets, capsules, oral liquids, drops, gels, granules, emulsions, creams, injections, eye drops, inhalants, sprays, aerosols or patches.

[0074] 32. The delivery kit according to any one of items 19 to 31, characterized in that the delivery system further comprises a polymer, preferably, the polymer has a group that can dissociate under physiological conditions and / or has 10 or more groups that can provide hydrogen donors or hydrogen acceptors, more preferably, the polymer is selected from the group consisting of:

[0075] (1) Cellulose polymers, such as hydroxypropyl methylcellulose (HPMC), low-substituted hydroxypropyl cellulose (L-HPC), hydroxypropyl methylcellulose acetate succinate (HPMCAS), sodium carboxymethylcellulose (CMC-Na), or hydroxyethyl cellulose;

[0076] (2) Synthetic polymers: such as polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyvinyl pyrrolidone (PVP), copolyvidone (PVP-VA64), polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (Soluplus), polyglutamic acid (PGA), polydopamine (PDA), polylactic acid (PLA), poly-L-lactic acid (PLLA), polylactic-co-glycolic acid (PLGA), sodium polyacrylate, siRNA and its derivatives (alkyl chains to improve lipophilicity), mRNA and its derivatives (alkyl chains to improve lipophilicity), or antisense oligonucleotides and their derivatives (alkyl chains to improve lipophilicity);

[0077] (3) polysaccharides and their derivatives (e.g., carboxymethylated, sulfonylated, phosphorylated, acylated or hydroxypropylated, cationic, anionic or other derivatives), such as carrageenan, guar gum, gum arabic, locust bean gum, konjac gum, agar, gelatin, pectin, gellan gum, hyaluronic acid (HA), aminodextran, chondroitin sulfate, dermatan sulfate, heparin, keratan sulfate, heparan sulfate, sodium alginate, propylene glycol alginate, agar, fucoidan, cyclodextrin and its derivatives, chitosan and its derivatives (e.g., acylated, carboxylated, alkylated and quaternized), soy protein, vegetable protein or bovine serum albumin; and

[0078] (4) Any combination of (1)-(3).

[0079] 33. The delivery kit according to any one of items 19 to 32, characterized in that the delivery system further comprises an additive selected from the group consisting of excipients, diluents, binders, disintegrants, lubricants, flavoring agents, pH regulators, osmotic pressure regulators, thickeners, plasticizers, colorants, film formers, preservatives or solvents, and any combination thereof.

[0080] 34. The delivery kit according to any one of items 19 to 33, for use selected from the group consisting of:

[0081] (1) For delivery of active ingredients to mammals (such as rodents, cattle, pigs, dogs, cats, primates, humans) or birds (such as chickens, ducks, geese);

[0082] (2) Use as solubilizer in agricultural chemistry, in particular in formulations containing pesticides, herbicides, fungicides or insecticides, in particular as crop protection agents for spray, pour-on or immersion mixtures;

[0083] (3) Use as a preservative, flavoring agent, or flavor enhancer for meat, vegetables, fruits, food, or beverages; and

[0084] (4) Used in cosmetics and fine chemical products to increase solubility, improve stability, and promote penetration.

[0085] In some embodiments, R1 in formula Ia can be a corresponding sugar residue in a naturally occurring stevia extract, such as shown in Table 1 below:

[0086] Table 1. The R1 group in formula Ia is the sugar group listed in the table

[0087] In some embodiments, the steviol glycoside derivatives and in vivo metabolites that can be used as delivery vehicles are selected from one or more combinations of the compounds in Table 2 below:

[0088] Table 2. Steviol glycoside derivatives used as carrier materials and their metabolites in vivo

[0089] The steviol glycoside derivatives used as delivery carrier materials of the present invention may be a single compound having the structure of Formula Ia and / or its hydrate and / or salt, or a mixture of multiple compounds having the structure of Formula Ia in any proportion and / or their hydrates.

[0090] The steviol glycoside derivatives that can be used as drug carrier materials described in the present invention have no restrictions on optical purity of the compound having the structure of Formula Ia, and the compound having the structure of Formula Ia has any enantiomeric excess value or any diastereomeric excess value.

[0091] Another aspect of the present invention discloses a method for preparing a compound of formula Ia, comprising the following steps:

[0092] (1) Step 1: Compound 1 is subjected to a hydrolysis reaction to obtain Compound 2, wherein when Compound 1 is in the form of a carboxylic acid, Step 2 is directly performed;

[0093] (2) Step 2: Protect the R1 group in compound 2 with a protecting group P to obtain compound 3;

[0094] (3) Step 3: subjecting compound 3 to a condensation reaction to obtain compound 4;

[0095] (4) Step 4: Compound 4 is subjected to a deprotection reaction to obtain a compound of formula Ia.

[0096] Wherein R1 or R2 in Compound 1, Compound 2, Compound 3 and Compound 4 is as defined in the compound represented by Structural Formula Ia.

[0097] The specific reaction process is as follows:

[0098] Step 1 is a non-limiting step. Most known steviol glycosides contain sugar ester bonds in their structures, such as stevioside, rebaudioside A (RBDS-A), dulcoside A (DCS-A), and rubusoside (RBSD), which are readily available from industrial production. Their structures are shown below. To obtain the compound of Formula Ia described herein, a hydrolysis reaction is required on a steviol glycoside containing a sugar ester bond.

[0099] If those skilled in the art have already known that steviol glycosides are carboxylic acid structures, such as the acid form (RBDS-C-Acid) of Steviolbioside (STVB), Rebaudioside B (RBDS-B), Rebaudioside C (RBDS-C), and the acid form (RBSD-Acid) of steviol or rubusoside as shown below, then this hydrolysis step is not required.

[0100] The hydrolysis conditions can be selected according to the ester hydrolysis conditions known in the art, for example, sodium hydroxide or potassium hydroxide can be used to carry out the reaction in water.

[0101] Without limitation, step 2 is the upper protection step. The selection of the protecting group can be based on suitable protecting groups known in the art for protecting sugars, such as silyl groups, acetyl groups, etc. In some embodiments, the sugar group is fully acetylated, such as RBDS-B-Ac, STVB-Ac, RBSD-Acid-Ac, DCS-A-Acid-Ac or STL-Ac, as shown in the following structure:

[0102] The conditions for adding the protecting group can be carried out according to the technical conditions known in the art. For example, the conditions for adding the protecting group of acetyl group can be carried out by using acetic anhydride and sodium acetate and carrying out the reaction under heating conditions.

[0103] Non-limitingly, step three is a condensation step, which requires compound 2 to react with a sugar, amine or alcohol with an R2 group to form a glycoside, amide or ester. The condensation reaction conditions can be carried out according to technical conditions known in the art. For example, the conditions for synthesizing amides can be found in "Large-Scale Amidations in Process Chemistry: Practical Considerations for Reagent Selection and Reaction Execution", Org. Process Res. Dev. 2022, 26, 1562-1689. In some embodiments, oxalyl chloride is first used to activate the carboxyl group of compound 2 to form an acyl chloride, and then reacted with an amino compound in the presence of alkaline conditions such as triethylamine.

[0104] Without limitation, step 4 is a deprotection step. The conditions for deprotection of the protecting group can be carried out according to technical conditions known in the art. For example, the conditions for deacetylation can be carried out using an alcohol solvent such as methanol or ethanol in the presence of a strong base such as potassium tert-butoxide, sodium methoxide, or sodium ethoxide.

[0105] Active ingredient delivery systems containing the delivery vehicles of the present invention can deliver the active ingredient more effectively than other compositions or compositions containing the active ingredient alone, and thus can administer to a subject a lower amount of the biological or chemical active agent than is used in existing unit dosage forms or delivery systems, while still achieving the same blood levels and / or therapeutic effect.

[0106] The active substance delivery system containing the delivery vehicle of the present invention has the effect of delivering the active agent to the selected biological system, target area and increasing or improving the bioavailability of the active agent than not using the delivery vehicle to deliver the active agent. Delivery can be improved by delivering more active agent to the target area over a period of time or within a specific time period (such as faster acting or delayed delivery) or within a period of time (such as sustained delivery) to deliver the active agent to the target area.

[0107] Generally, the weight ratio of delivery vehicle to active agent ranges from about 1: 100 to about 300: 1. This weight ratio will vary depending upon the active agent and the particular indication for which the active agent is to be administered.

[0108] In some embodiments, the additive is a pharmaceutically acceptable additive, such as those described in Remington: The Science and Practice of Pharmacy (Adeboye Adejare, ed., 23rd edition, 2020, Academic Press), and Pharmacy (Fang Liang, ed., 8th edition, 2016, People's Medical Publishing House), both of which are incorporated herein by reference.

[0109] Advantageous Effects of the Invention

[0110] The stevioside derivatives provided by the present invention have better permeability than existing steviosides, and can carry substances that need to pass through biological membranes to achieve transmembrane transport; the hyperbranched structure can interact with various substances through multiple sites to form a relatively stable assembly complex, avoiding the destruction of the complex by the contents of the digestive tract during the transport process. The stevioside derivatives provided by the present invention have good lipophilicity while maintaining the hydrophilicity of stevioside compounds, and are more likely to self-assemble into complexes with active ingredients of different physicochemical properties, thereby achieving delivery in different scenarios; therefore, the carrier provided by the present invention can improve the stability of nanomedicines during production and storage; according to different delivery scenarios, different forms of in situ assembly can be achieved under the action of enzymes or microorganisms; it is derived from existing glycoside substances or metabolites in the body, has a certain water solubility and is endowed with a certain lipophilicity or dissociability, and has better biocompatibility and safety. Compared to high-molecular-weight polymers, steviol glycoside derivatives possess the properties of nanocarriers, allowing them to be absorbed into the bloodstream along with the active ingredient and reach the target area (such as a target organ, tissue, or cell). However, due to their large molecular weight, the active ingredient must be released before absorption. Therefore, the steviol glycoside derivatives provided by the present invention are excellent active ingredient delivery vehicles. BRIEF DESCRIPTION OF THE DRAWINGS

[0111] FIG1 is a mass spectrum of the RBDS-B intermediate prepared in Example 1 under LC-MS / MS negative ion mode;

[0112] Figure 2 is the RBDS-B intermediate obtained in Example 1 1 H-NMR spectrum;

[0113] FIG3 is a mass spectrum of the compound RBDS-B-His obtained in Example 2 under LC-MS / MS negative ion mode;

[0114] Figure 4 shows the compound RBDS-B-His obtained in Example 2 1 H-NMR spectrum;

[0115] FIG5 is a mass spectrum of the compound RBDS-B-APL obtained in Example 3 under LC-MS / MS negative ion mode;

[0116] Figure 6 shows the compound RBDS-B-APL obtained in Example 3 1 H-NMR spectrum;

[0117] FIG7 is a mass spectrum of the compound STVB obtained in Example 4 under LC-MS / MS negative ion mode;

[0118] FIG8 is a graph showing the compound STVB obtained in Example 4. 1 H-NMR spectrum;

[0119] FIG9 is a mass spectrum of the compound STVB-Arga obtained in Example 5 under LC-MS / MS negative ion mode;

[0120] Figure 10 is the compound STVB-Arga obtained in Example 5 1 H-NMR spectrum;

[0121] FIG11 is a mass spectrum of the compound STVB-His obtained in Example 6 under LC-MS / MS negative ion mode;

[0122] Figure 12 shows the compound STVB-His obtained in Example 6 1 H-NMR spectrum;

[0123] FIG13 is a mass spectrum of the compound STVB-APL obtained in Example 7 under LC-MS / MS negative ion mode;

[0124] Figure 14 shows the compound STVB-APL obtained in Example 7 1 H-NMR spectrum;

[0125] Figure 15 is the compound RBDS-B-Na obtained in Example 8 1 H-NMR spectrum;

[0126] FIG16 is a LC-MS / MS mass spectrum of the compound STVB-CPP-Arg8 obtained in Example 9 under positive and negative ion modes;

[0127] Figure 17 shows the compound STVB-CPP-Arg8 obtained in Example 9 1 H-NMR spectrum;

[0128] FIG18 is a mass spectrum of the compound STL-CPP-Arg2 obtained in Example 10 under LC-MS / MS positive ion mode;

[0129] Figure 19 shows the compound STL-CPP-Arg2 obtained in Example 10 1 H-NMR spectrum;

[0130] FIG20 is a mass spectrum of the compound STL-CPP-Arg4 obtained in Example 11 under LC-MS / MS positive ion mode;

[0131] Figure 21 shows the compound STL-CPP-Arg4 obtained in Example 11 1 H-NMR spectrum;

[0132] FIG22 is a mass spectrum of the compound STL-CPP-Arg8 obtained in Example 12 under LC-MS / MS positive ion mode;

[0133] Figure 23 shows the compound STL-CPP-Arg8 obtained in Example 12 1 H-NMR spectrum;

[0134] FIG24 is a mass spectrum of the compound RBSD-Arga obtained in Example 13 under LC-MS / MS positive and negative ion modes;

[0135] Figure 25 is the compound RBSD-Arga obtained in Example 13 1 H-NMR spectrum;

[0136] Figure 26 is a typical spectrum of Example 17 STVB-CPP-Arg8 simulated intestinal fluid (trypsin) enzymatic hydrolysis for 1 h

[0137] Figure 27 is a comparison of the permeability of the new derivative of Example 19 and Comparative Example 1.

[0138] Figure 28-1 is a mass spectrometry TIC diagram of STVB-CPP-Arg metabolism in human intestinal flora for 8 hours in Example 20;

[0139] Figure 28-2 is the negative ion mode mass spectrum of each metabolite peak of STVB-CPP-Arg metabolized by human intestinal flora for 8 hours in Example 20. DETAILED DESCRIPTION

[0140] The raw materials, reagents and solvents used in the present invention are not particularly limited, and commercially available conventional raw materials, reagents and solvents can be used.

[0141] Instruments and methods used to collect data:

[0142] Acquisition of H NMR spectra ( 1 H NMR) data, the instrument model used is: Bruker AVANCE600, resonance frequency: 600 MHz, solvent used is deuterated DMSO.

[0143] The high-resolution mass spectrometry data of the present invention was detected using a Waters Xevo TQ-S triple quadrupole mass spectrometer, and the liquid chromatography conditions were as follows:

[0144] The chromatographic column was Agilent ZOBAX SB-C18, 2.1 × 50 mm, 1.8 μm;

[0145] Mobile phase A: 0.1% formic acid-water solution; Mobile phase B: 0.08% formic acid-acetonitrile solution;

[0146] Detection wavelength: 210nm;

[0147] Flow rate: 0.7 mL / min;

[0148] Injection volume: 5 μL;

[0149] Column temperature: 60°C;

[0150] The gradient elution program is shown in Tables 3 and 4.

[0151] Table 3. Gradient elution procedures for RSDB-B-Na, STVB-Arga, STVB-His, STVB-CPP, and STVB-APL

[0152] Table 4. Gradient elution program of RBDS-B-His and RBDS-B-APL

[0153] The mass spectrometry detection conditions were as follows: capillary voltage: 2.80 kV; drying gas temperature: 350°C; positive / negative ion scanning mode, and ESI ion source.

[0154] The content of RBDS-B or STVB sodium salt was determined by potentiometric titration.

[0155] Titration was performed using a Metrohm 916 Ti-Touch potentiometric titrator. Dissolve approximately 0.4 g of this product in 50 ml of ethanol-water (50:50) and titrate using 0.1 mol / L hydrochloric acid solution according to the potentiometric titration method (Chinese Pharmacopoeia 2020 Edition, Part IV, General Rules 0701). The titration result was corrected using a blank test. Each 1 ml of 0.1 mol / L hydrochloric acid solution is equivalent to 2.30 mg of sodium. The theoretical sodium content of this product, calculated as anhydrous, is 2.78%.

[0156] The purity of each derivative was tested by high performance liquid chromatography, and the chromatographic conditions are shown in Table 5 below:

[0157] Table 5. Chromatographic conditions

[0158] The purity of the main peak was calculated using the principal component self-reference method.

[0159] It should be noted that the protection scope of the meaning or significance of the numerical values ​​or numerical endpoints involved in the technical solution of the present invention is not limited to the numbers themselves. Those skilled in the art will understand that they include those allowable error ranges that have been widely accepted in the art, such as experimental errors, measurement errors, statistical errors and random errors, etc., and these error ranges are all included in the scope of the present invention.

[0160] In order to further illustrate the present invention, specific examples will be given below, but the following examples do not constitute any limitation to the scope of protection of the present invention.

[0161] Example 1. Synthesis of RBDS-B-Ac-Cl

[0162] In a 1 L three-necked flask, 100 g of rebaudioside A (RBDS-A) and 720 ml of 5% potassium hydroxide aqueous solution were added, the reaction system was heated to 85° C. to 90° C., and the reaction was stirred for 4 hours. The reaction system was cooled to room temperature and the pH of the system was adjusted to 5-6 with 6N hydrochloric acid. Solid precipitated and was filtered. The filter cake was rinsed twice with 400 mL of purified water. The filter cake was collected and dried under reduced pressure at 40° C. to obtain 80 g of rebaudioside B (RBDS-B) as a light brown solid. The LC-MS / MS liquid chromatography retention time was approximately 26.6 min; the mass-to-charge ratio of the negative ion by LC-MS / MS was 803.8409 (MH). - , which is consistent with the theoretical precise molecular weight of RBDS-B. The high-resolution mass spectrometry negative ion mode spectrum is shown in Figure 1. 1 H NMR(600MHz,d-DMSO)δ11.957(s,1H),5.610-5.601(d,J=5.4,1H),5.156-5.135(m,2H ),5.084-5.076(d,J=4.8,1H),5.021-5.012(d,J=5.4,1H),4.846(s,2H),4.767(s,1H) ,4.672-4.659(d,J=7.8,1H),4.596-4.579(m,1H),4.509-4.491(m,2H),4.442-4.408 (m,2H),4.033(m,1H),3.714-3.545(m,4H),3.482-3.392(m,4H),3.233-2.986(m,9H), 2.945-2.918 (m, 1H), 2.078-1.932 (m, 4H), 1.874-1.698 (m, 6H), 1.529-1.344 (m, 6H), 1.112 (s, 3H), 1.025-0.928 (m, 3H), 0.844 (s, 3H), 0.807-0.767 (m, 1H), see Figure 2 for details, HPLC purity 93.78%.

[0163] In a 2L three-necked flask, 72g of rebaudioside B (RBDS-B), 180g of acetic anhydride and 1.8g of sodium acetate were added, the reaction system was heated to 140°C, and the reaction was stirred for 6 hours. The reaction system was cooled to room temperature, 1.6mL of dichloromethane was added, and the organic phase was separated. The organic phase was washed three times with 800mL of purified water, and the organic phase was collected and dried under reduced pressure at 40°C to obtain 100g of RBDS-B-Ac as a light yellow solid.

[0164] In a 1L three-necked flask, add 40g of RBDS-B-Ac and 600mL of dichloromethane, stir the reaction system until dissolved, add 1 drop of N,N-dimethylformamide, and then add 6.2g of oxalyl chloride dropwise at room temperature. After the addition is complete, stir and react at room temperature for 4 hours. The reaction system is dried under reduced pressure at 40°C to obtain 42g of RBDS-B-Ac-Cl as a light yellow solid.

[0165] Example 2. Synthesis of RBDS-B-His

[0166] In a 2L three-necked flask, 11.48g of L-histidamide hydrochloride, 840mL of dichloromethane, and 17.5g of triethylamine were added, and then 21g of RBDS-B-Ac-Cl was added in batches. After the addition, the reaction system was stirred at room temperature for 4 hours. The reaction system was washed three times with 350mL of purified water, and the organic phase was collected and dried under reduced pressure at 40°C to obtain 18.2g of RBDS-B-His-Ac as a yellow solid.

[0167] In a 2L three-necked flask, 18.2g of RBDS-B-His-Ac, 700mL of methanol, and 18.9g of potassium tert-butoxide were added. The reaction system was stirred at room temperature for 4 hours. The reaction system was dried under reduced pressure at 40°C to obtain a crude RBDS-B-His product. After reverse-phase preparative separation, 9.1g of RBDS-B-His was obtained as an off-white solid. The main peak retention time in the LC-MS / MS liquid chromatogram was approximately 20.6min, and the mass-to-charge ratio of the main peak in the positive / negative ion mode was 940.8148 (M+H), respectively. + / 938.9222(MH) - , which is consistent with its theoretical precise molecular weight. The high-resolution mass spectrometry negative ion mode spectrum is shown in Figure 3. 1 H NMR(600MHz,d-DMSO)δ8.239(s,1H),7.718-7.570(m,1H),7.233-7.186(m,1H),7.01 4-6.982(m,1H),6.873-6.766(m,1H),5.092-4.989(m,2H),4.731-4.395(m,7H),3.70 9-3.409 (m, 13H), 3.280-2.871 (m, 15H), 2.079-1.678 (m, 8H), 1.508-1.297 (m, 8H), 1.073-0.983 (m, 3H), 0.928-0.907 (m, 2H), 0.731-0.633 (m, 4H), see Figure 4 for details; HPLC purity is 99.09%.

[0168] Example 3. Synthesis of RBDS-B-APL

[0169] In a 100 mL three-necked flask, 0.20 g of S-1-amino-2-propanol, 50 mL of dichloromethane, and 0.6 g of triethylamine were added, and then 1 g of RBDS-B-Ac-Cl was added in batches. After the addition, the reaction system was stirred at room temperature for 2 hours. The reaction system was washed three times with 30 mL of purified water, and the organic phase was collected and dried under reduced pressure at 40 ° C to obtain 0.9 g of RBDS-B-APL-Ac as a foamy light yellow solid.

[0170] In a 100 mL three-necked flask, 0.9 g of RBDS-B-APL-Ac, 50 mL of methanol, and 0.89 g of potassium tert-butoxide were added. The reaction system was stirred at room temperature for 4 hours. The reaction system was dried under reduced pressure at 40 ° C to obtain a crude RBDS-B-APL product. After reverse phase preparative separation, 0.4 g of RBDS-B-APL was obtained as an off-white solid. The LC-MS / MS liquid chromatogram showed that the retention time of the main peak was about 24.5 min, and the mass-to-charge ratio of the main peak in positive / negative ion mode was 861.7228 (M+H) respectively. + / 860.0262(MH) - And the combined peak mass-to-charge ratio is 905.7061 (M+2Na-H) - , which is consistent with the theoretical precise molecular weight of the derivative. The high-resolution mass spectrometry negative ion mode spectrum is shown in Figure 5; 1 H NMR (600 MHz, d-DMSO+D2O) δ 5.093 (s, 1H), 4.758 (s, 1H), 4.678-4.665 (d, J-7.8 Hz, 1H), 4.476-4.458 (m, 2H), 3.697-3.570 (m, 5H), 3.475-3.339 (m, 4H), 3.219-2.914 (m, 12H), 2.083-1.943 (m, 4H), 1.814-1.601 (m, 6H), 1.558-1.324 (m, 6H), 1.035-0.878 (m, 9H), 0.788-0.750 (m, 4H), see Figure 6 for details; HPLC purity was 97.23%.

[0171] Example 4. Synthesis of STVB-Ac-Cl

[0172] In a 2L three-necked flask, 100g of stevioside (STVS) and 800ml of 5% potassium hydroxide aqueous solution were added, the reaction system was heated to 60°C, stirred and reacted for 6 hours, the reaction system was cooled to room temperature, and the pH value of the system was adjusted to 5-6 with 6N hydrochloric acid. Solids precipitated and were filtered. The filter cake was rinsed twice with 400mL of purified water, and the filter cake was collected and dried under reduced pressure at 40°C to obtain 73g of STVB as a white solid. The main peak retention time of the LC-MS / MS liquid phase was about 27.2min, and the mass-to-charge ratios of the main peak in the positive and negative ion modes were 660.3461 (M+H2O), respectively. + / 641.0961(MH) - , the high-resolution mass spectrometry negative ion mode spectrum is shown in Figure 7, 1 H NMR (600MHz, d-DMSO+D2O) δ5.099(s,1H),4.745(s,1H),4.470-4.457(d,J=7.8Hz,1H),4.372-4.359(d,J=7. 8Hz,1H),3.609-3.570(m,2H),3.492-3.428(m,2H),3.400-3.370(m,1H),3.232-3.204(m,1H),3.184-3.117 (m, 3H), 3.060-2.985 (m, 3H), 2.210-1.966 (m, 4H), 1.882-1.696 (m, 6H), 1.520-1.467 (m, 3H), 1.399-1.316 (m, 3H), 1.090 (s, 3H), 0.991-0.967 (m, 1H), 0.923-0.885 (m, 5H), 0.794-0.747 (m, 1H), see Figure 8 for details; HPLC purity is 98.26%.

[0173] In a 1L three-necked flask, 36g of STVB, 84g of acetic anhydride and 11g of sodium acetate were added, the reaction system was heated to 140°C, and the reaction was stirred for 6 hours. The reaction system was cooled to room temperature, 800mL of dichloromethane was added, the organic phase was separated, and the organic phase was washed three times with 400mL of purified water. The organic phase was collected and dried under reduced pressure at 40°C to obtain 47g of STVB-Ac as a light brown solid.

[0174] In a 500mL three-necked flask, add 47g of STVB-Ac and 600mL of dichloromethane, stir the reaction system until dissolved, add 1 drop of N,N-dimethylformamide, and then add 9.3g of oxalyl chloride dropwise at room temperature. After the addition is complete, stir and react at room temperature for 4 hours. The reaction system is dried under reduced pressure at 40°C to obtain 22g of STVB-Ac-Cl as a light brown foamy solid.

[0175] Example 5. Synthesis of STVB-Arga

[0176] In a 250 mL three-necked flask, 4.5 g of L-arginine amide dihydrochloride, 150 mL of N,N-dimethylformamide, and 4.5 g of triethylamine were added, and then 6.0 g of STVB-Ac-Cl was added in batches. After the addition, the reaction system was stirred at room temperature for 6 hours. The reaction system was dried under reduced pressure at 40 ° C to obtain 19.5 g of STVB-Arga-Ac as a foamy light yellow solid.

[0177] In a 500 mL three-necked flask, 19.5 g of STVB-Arga-Ac, 150 mL of methanol, and 5.4 g of potassium tert-butoxide were added. The reaction system was stirred at room temperature for 4 hours. The reaction system was then dried under reduced pressure at 40°C to obtain a crude STVB-Arga product. After reverse phase preparative separation, 2.1 g of STVB-Arga was obtained as an off-white solid. The main peak retention time in the LC-MS / MS liquid phase spectrum was approximately 21.5 min, and the mass-to-charge ratios of the main peak positive and negative ions were 797.8373 (M+H), respectively. + / 796.1407(MH) - and the combined peak mass-to-charge ratio 841.9503 (M+2Na-H) - , which is consistent with the theoretical precise molecular weight of the main peak. The high-resolution mass spectrometry negative ion mode spectrum is shown in Figure 9. 1 H NMR(600MHz,d-DMSO)δ8.721(s,1H),7.588-7.534(m,4H),7.074(s,1H),6.884-6.8 43(m,1H),5.806(s,1H),5.450(s,1H),5.052-5.032(m,3H),4.735(s,1H),4.516-4 .273 (m, 5H), 3.625-3.386 (m, 5H), 3.236-2.981 (m, 10H), 2.141-1.966 (m, 3H), 1.880-1.356 (m, 18H), 1.088-0.913 (m, 5H), 0.800-0.789 (m, 4H), see Figure 10 for details, HPLC purity is 97.82%.

[0178] Example 6. Synthesis of STVB-His

[0179] In a 1 L three-necked flask, 5.71 g of L-histidamide hydrochloride, 200 mL of N,N-dimethylformamide, and 3.0 g of triethylamine were added, and then 8.0 g of STVB-Ac-Cl was added in batches. After the addition, the reaction system was stirred at room temperature for 4 hours. The reaction system was dried under reduced pressure at 40 ° C to obtain 29.1 g of STVB-His-Ac as a foamy light yellow solid.

[0180] In a 500 mL three-necked flask, 29.1 g of STVB-His-Ac, 200 mL of methanol, and 7.2 g of potassium tert-butoxide were added. The reaction system was stirred at room temperature for 4 hours. The reaction system was dried under reduced pressure at 40 ° C to obtain a crude STVB-His product. After reverse phase preparative separation, 3.2 g of STVB-His was obtained as an off-white solid. The main peak retention time in LC-MS / MS liquid chromatography was about 20.7 min, and the mass-to-charge ratios of the main peak positive and negative ions were 778.9135 (M+1), respectively. + / 777.0210(MH) - and combined peak 823.0913 (M+2Na-H) - , the high-resolution mass spectrometry negative ion mode spectrum is shown in Figure 11; 1 H NMR(600MHz,d-DMSO)δ11.818(s,1H),7.811-7.532(m,1H),7.211-7.191(m,1H),7.0 81-6.893(m,2H),6.769-6.611(m,1H),5.774-5.309(m,3H),5.106-4.738(m,5H),4.6 17-4.534 (m, 2H), 4.392-4.345 (m, 2H), 3.657-3.405 (m, 5H), 3.287-2.865 (m, 9H), 2.044-1.242 (m, 16H), 1.031-0.853 (m, 6H), 0.640-0.495 (m, 4H), see Figure 12 for details, HPLC purity is 97.64%.

[0181] Example 7. Synthesis of STVB-APL

[0182] In a 100 mL three-necked flask, 0.25 g of S-1-amino-2-propanol (APL), 50 mL of dichloromethane, and 0.7 g of triethylamine were added, and then 1 g of STVB-Ac-Cl was added in batches. After the addition, the reaction system was stirred at room temperature for 2 hours. The reaction system was washed three times with 30 mL of purified water, and the organic phase was collected and dried under reduced pressure at 40 ° C to obtain 1.4 g of STVB-APL-Ac as a foamy light yellow solid.

[0183] In a 100 mL three-necked flask, 1.2 g of STVB-APL-Ac, 20 mL of methanol, and 1.2 g of potassium tert-butoxide were added. The reaction system was stirred at room temperature for 4 hours. The reaction system was dried under reduced pressure at 40 ° C to obtain RBDS-APL crude product. After reverse phase preparative separation, 0.5 g of STVB-APL was obtained as an off-white solid. The main peak retention time of the LC-MS / MS liquid chromatogram was about 24.6 min, and the mass-to-charge ratio of the main peak in the positive and negative ion modes was 699.8901 (M+H) respectively. + 、1400.4702(2M+H) + / 698.1935(MH) - 、744.9809(M+Na-H) - 、1397.4629(2M-H) - 、1443.6785(2M+2Na+H) + , which is consistent with the theoretical precise molecular weight. The high-resolution mass spectrometry negative ion mode spectrum is shown in Figure 13. 1 H NMR(600MHz,d-DMSO)δ6.914-6.895(t,J=5.4Hz,1H),5.699-5.663(d,J=21.6Hz,1H),5.365(s, 1H),5.072-4.691(m,5H),4.497-4.476(m,1H),4.414-4.328(m,2H),4.204-4.166(m,1H),3.69 5-3.657 (m, 1H), 3.609-3.593 (m, 2H), 3.491-3.373 (m, 4H), 3.238-2.975 (m, 9H), 2.120-1.966 (m, 3H), 1.854-1.688 (m, 5H), 1.620-1.344 (m, 8H), 1.060-0.751 (m, 13H), see Figure 14 for details, HPLC purity is 96.92%.

[0184] Example 8. Synthesis of RBDS-B-Na

[0185] In a 500mL three-necked flask, 28.00g of RBDS-B was dispersed in 280mL of purified water. 1.39g of sodium hydroxide solid was added in batches while controlling the temperature to be less than 40°C. After the addition, the mixture was stirred until the system dissolved. The slightly insoluble matter was removed by filtration. 560ml of anhydrous ethanol was added to the filtrate, and then the temperature was lowered to -10°C and stirred for crystallization for 3 hours. The mixture was filtered and the filter cake was collected and dried under reduced pressure at 40°C to obtain 22g of RBDS-B-Na as an off-white solid. 1H NMR(600 MHz,d-DMSO)δ6.367(s,2H),5.682(m,4H),5.120(s,1H),4.898(s,1H),4.775-4.737(m,2H),4.640 (s,1H),4.510(s,1H),4.457-4.432(m,2H),4.357-4.235(m,1H),3.685-3.556(m,4H),3.491-3.40 2(m,4H),3.246-3.046(m,10H),2.240-2.220(m,1H),2.058-1.858(m,6H),1.722-1.646(m,3H),1.465-1.204(m,6H),0.979-0.959(m,6H),0.847-0.680(m,4H), the sodium content was determined to be 2.77% (n=3), which is about 98.2% of the theoretical value. 1 The H NMR spectrum is shown in Figure 15 , and the HPLC purity was 96.7%.

[0186] Example 9. Synthesis of STVB-CPP-Arg8

[0187] The CPP is octaarginine (Arg8).

[0188] In a 250 mL three-necked flask, 3.18 g of octaarginine (cell-penetrating peptide CPP-Arg8), 50 mL of N,N-dimethylformamide, 1.35 g of diisopropylethylamine, and then 2.0 g of STVB-Ac-Cl were added. After the addition, the reaction system was stirred at room temperature for 12 hours. The reaction system was dried under reduced pressure at 40 ° C to obtain 9.2 g of STVB-Ac-CPP-Arg8 as a foamy light yellow solid.

[0189] In a 100 mL three-necked flask, 9.2 g of STVB-Ac-CPP-Arg8, 25 mL of methanol, 30 mL of N,N-dimethylformamide, and 2.0 g of sodium methoxide were added. The reaction system was stirred at room temperature for 8 hours. The reaction system was then dried under reduced pressure at 40°C to obtain crude STVB-CPP-Arg8. After preparative separation by reverse-phase HPLC, 1.8 g of STVB-CCP-Arg8 was obtained as an off-white solid with an HPLC purity of 92.33%. The main peak retention time of the LC-MS / MS liquid chromatogram was approximately 5.8 min, and the mass-to-charge ratio of the main peak in positive and negative ion modes was 946.0657 (M / 2+H), respectively. + 、993.1732(M / 2+2Na+H) + / 944.1567(M / 2-H)- 、990.5209(M / 2+Na-H) - , and its absolute error with the theoretical precise molecular weight is less than 5ppm. The high-resolution mass spectrometry positive ion mode spectrum is shown in Figure 16; 1H NMR (600MHz, d-DMSO) δ11.52(s,1H),8.41(s,8H),7.93(s,8H),6.55(s,16H),5.38-5.34(d,J=24Hz,1H),5.23(s,1H),5.15-4.82(m,5H),4.70-4.64(m,3H),4.53-4.50(d,J=18Hz,1H),4.27-4.22(m,7H),4.04-3.94(m,3H),3.76-3.72(m,1H),3.71-3.67(m,2H),3. 65-3.62(m,2H),3.44-3.40(m,1H),3.39-3.34(m,2H),3.28-3.24(m,2H),3.14-3.12(m,16H),2.59(s,8H),2.17-2.15(d,J=12Hz,1H),2.04-2.02(m,2H),1.86-1.81(m,1H),1.78-1.41(m,46H),1.30-1.26(m,1H),1.07(s,3H),1.02-1.00(m,1H),0.95(s,3H), NMR 1 The HNMR spectrum is shown in Figure 17.

[0190] Example 10. Synthesis of STL-CPP-Arg2

[0191] In a 500 mL three-necked flask, 50 g of steviol (STL), 160 g of acetic anhydride and 32.0 g of sodium acetate were added, the reaction system was heated to 140 ° C, and the reaction was stirred for 12 hours. The reaction system was cooled to room temperature, 600 mL of dichloromethane was added, and the organic phase was washed three times with 200 mL of purified water. The organic phase was collected and dried under reduced pressure at 40 ° C to obtain 65 g of STL-Ac as a light yellow foamy solid.

[0192] In a 1000mL three-necked flask, add 65g of STL-Ac and 650mL of dichloromethane, stir the reaction system until dissolved, add 1 drop of N,N-dimethylformamide, and then add 45g of oxalyl chloride dropwise at room temperature. After the addition is complete, stir and react at room temperature for 8 hours. The reaction system is dried under reduced pressure at 40°C to obtain 60g of STL-Ac-Cl as a light yellow foamy solid.

[0193] In a 100 mL three-necked flask, 2.5 g of 2-polyarginine, 50 mL of N-methylpyrrolidone, and 3.5 g of diisopropylethylamine were added, and then 2.0 g of STL-Ac-Cl was added in batches. After the addition was completed, the reaction system was stirred at room temperature for 16 hours, and then 15 mL of methanol and 1.0 g of sodium methoxide were added. The reaction system continued to stir at room temperature for 16 hours to obtain a solution of STL-CPP-Arg2 crude product in N-methylpyrrolidone. After reverse phase preparative separation, 1.6 g of STL-CPP-Arg2 was obtained as an off-white solid. The main peak retention time of the LC-MS / MS liquid chromatogram was about 9.8 min, and the mass-to-charge ratio of the main peak in the positive ion mode was 631.4226 (M+H) + 、1261.8796(2M+H) + 、Mass-to-charge ratio in negative ion mode: 629.4198 (MH) - 、1259.4659(2M-H) - The absolute error with its theoretical precise molecular weight is no more than 5ppm. The high-resolution mass spectrometry positive ion mode spectrum is shown in Figure 18. 1 H NMR (600 MHz, d-DMSO+D2O) δ 4.959 (m, 1H), 4.908 (m, 1H), 4.217-4.199 (m, 1H), 3.991-3.970 (m, 1H), 3.140-3.118 (m, 4H), 2.142-1.990 (m, 3), 1.751-1.442 (m, 23H), 1.297-1.259 (m, 1H), 1.074 (s, 3H), 1.019-1.009 (m, 1H), 0.954 (s, 3H), see Figure 19 for details. The HPLC purity was 96.75%.

[0194] Example 11. Synthesis of STL-CPP-Arg4

[0195] In a 100 mL three-necked flask, 5.0 g of 4-polyarginine, 50 mL of N-methylpyrrolidone, and 3.5 g of diisopropylethylamine were added, and then 2.0 g of STL-Ac-Cl was added in batches. After the addition, the reaction system was stirred at room temperature for 17 hours, and then 15 mL of methanol and 1.0 g of sodium methoxide were added. The reaction system continued to stir at room temperature for 17 hours to obtain a solution of STL-CPP-Arg4 crude product in N-methylpyrrolidone. After reverse phase preparative separation, 2.5 g of STL-CPP-Arg4 was obtained as an off-white solid. The retention time of the main peak of the LC-MS / MS liquid chromatogram was about 6.7 min. The mass-to-charge ratio of the main peak in the positive ion mode was 943.6235 (M+H)+ and the mass-to-charge ratio in the negative ion mode was 941.6229 (MH)-, and the absolute error with its theoretical precise molecular weight was no more than 5 ppm. The high-resolution mass spectrometry positive ion mode spectrum is shown in Figure 20. 1 H NMR (600 MHz, d-DMSO+D2O) δ 4.96 (m, 1H), 4.91-4.90 (m, 1H), 4.27-4.22 (m, 3H), 3.99-3.97 (m, 1H), 3.15-3.11 (m, 8H), 2.13-1.96 (m, 3H), 1.78-1.43 (m, 31H), 1.43-1.40 (m, 1H), 1.07-1.02 (m, 4H), 0.94 (s, 3H), see Figure 21 for details. The HPLC purity was 93.84%.

[0196] Example 12. Synthesis of STL-CPP-Arg8

[0197] In a 100 mL three-necked flask, 5.0 g of octaarginine, 50 mL of N-methylpyrrolidone, and 1.7 g of diisopropylethylamine were added, and then 1.0 g of STL-Ac-Cl was added in batches. After the addition, the reaction system was stirred at room temperature for 18 hours, and then 15 mL of methanol and 0.8 g of sodium methoxide were added. The reaction system continued to stir at room temperature for 18 hours to obtain a solution of STL-CPP-Arg8 crude product in N-methylpyrrolidone. After reverse phase preparative separation, 1.8 g of STL-CPP-Arg8 was obtained as an off-white solid. The main peak retention time of LC-MS / MS was about 6.5 min, and the mass-to-charge ratio of the main peak in positive ion mode was 784.5119 (M / 2+H). + 、801.5553(M / 2+H2O) + , mass-to-charge ratio 782.5127 (M / 2-H) in negative ion mode - , and its absolute error with its theoretical precise molecular weight does not exceed 5ppm. The high-resolution mass spectrometry positive ion mode spectrum is shown in Figure 22; 1H NMR (600 MHz, d-DMSO+D2O) δ 5.08 (m, 1H), 4.96 (m, 1H), 4.37-4.35 (m, 7H), 4.27-4.22 (m, 1H), 3.31-3.26 (m, 16H), 2.19-1.99 (m, 3H), 1.76-1.14 (m, 47H), 1.39-1.36 (m, 1H), 1.11 (s, 3H), 1.00-0.95 (m, 4H), see Figure 23 for details. The HPLC purity was 92.13%.

[0198] Example 13. Synthesis of RBSD-Arga

[0199] In a 250mL three-necked flask, add 10g of rubusoside (RBSD) and 100ml of 10% sodium hydroxide aqueous solution, heat the reaction system to 85°C to 90°C, keep stirring and react for 6 hours, cool the reaction system to room temperature, adjust the pH value of the system to 5-6 with 6N hydrochloric acid, filter the solid precipitated, rinse the filter cake twice with 100mL of purified water, collect the filter cake and dry it under reduced pressure at 40°C to obtain 5.6g of RBSD-Acid as a brown solid.

[0200] In a 500 mL three-necked flask, 5.6 g RBSD-Acid, 25 g acetic anhydride and 1.5 g sodium acetate were added, the reaction system was heated to 140 ° C, and the reaction was stirred for 8 hours. The reaction system was cooled to room temperature, 400 mL of dichloromethane was added, and the organic phase was washed three times with 200 mL of purified water. The organic phase was collected and dried under reduced pressure at 40 ° C to obtain 8.7 g of RBSD-Acid-Ac as a brown solid.

[0201] In a 500 mL three-necked flask, add 8.7 g of RBSD-Acid-Ac and 300 mL of dichloromethane, stir the reaction system until dissolved, add 1 drop of N,N-dimethylformamide, and then add 3.4 g of oxalyl chloride dropwise at room temperature. After the addition is complete, stir and react at room temperature for 8 hours. The reaction system is dried under reduced pressure at 40 ° C to obtain 6.5 g of RBSD-Ac-Cl as a foamy brown solid.

[0202] In a 100 mL three-necked flask, 1.1 g of L-arginine amide dihydrochloride, 50 mL of N,N-dimethylformamide, and 3.0 g of diisopropylethylamine were added, and then 2.0 g of RBSD-Ac-Cl was added in batches. After the addition, the reaction system was stirred at room temperature for 6 hours. The reaction system was dried under reduced pressure at 65 ° C to obtain 4.5 g of RBSD-Ac-Agra as a foamy brown solid.

[0203] In a 100 mL three-necked flask, 4.5 g of RBSD-Ac-Agra, 15 mL of methanol, 50 ml of N-methylpyrrolidone and 1.8 g of potassium tert-butoxide were added. The reaction system was stirred at room temperature for 8 hours. The reaction system was evaporated under reduced pressure at 40 ° C to obtain a solution of crude RBSD-Arga in N-methylpyrrolidone. After reverse phase preparative separation, 0.8 g of RBSD-Arga was obtained as an off-white solid. The main peak retention time of the LC-MS / MS liquid chromatogram was about 22.2 min, and the mass-to-charge ratio of the main peak in negative ion mode was 634.3876 (MH) - The mass-to-charge ratio in positive ion mode is 636.3656 (M+H) + , and its absolute error with its theoretical precise molecular weight does not exceed 5ppm. The high-resolution mass spectrometry positive ion mode spectrum is shown in Figure 24; 1 H NMR (600 MHz, d-DMSO+D2O) δ 5.27 (s, 1H), 4.86 (s, 1H), 4.77-4.75 (m, 1H), 4.13-4.08 (m, 1H), 3.67-3.10 (m, 8H), 2.21-2.04 (m, 3H), 1.90-1.30 (m, 19H), 1.28-1.27 (m, 1H), 1.13-1.07 (m, 4H), 0.98 (s, 3H), see Figure 25 for details. The HPLC purity was 97.32%.

[0204] Example 14. Moisture absorption test

[0205] Appropriate amounts of the above derivatives were weighed and tested for hygroscopicity according to the "Guiding Principles for Hygroscopicity of Drugs" in General Chapter 9103 of Part IV of the 2020 Chinese Pharmacopoeia. The test conditions were an artificial climate chamber maintained at 25°C ± 1°C and a relative humidity of 80% ± 2%. The test lasted 24 hours, and the weight was accurately weighed at the end of the test. The results are shown in Table 6.

[0206] Table 6. Hygroscopicity test results of various derivatives

[0207] The above derivatives were tested in a stability sample box at 25°C / RH80% for 24 hours. According to the description of hygroscopic characteristics and the definition of hygroscopic weight gain in the General Chapter 9103 of the 2020 edition of the Chinese Pharmacopoeia, Part IV: "Guiding Principles for Hygroscopic Tests of Drugs":

[0208] Highly hygroscopic: weight gain due to moisture absorption is not less than 15%;

[0209] Hygroscopic: weight gain due to moisture absorption is less than 15%, but not less than 2%;

[0210] Slightly hygroscopic: weight gain due to moisture absorption is less than 2%, but not less than 0.2%;

[0211] No or almost no hygroscopicity: weight gain due to moisture is less than 0.2%.

[0212] Hygroscopicity test results show that this product is extremely hygroscopic. RBDS-CPP-Arg8 is extremely hygroscopic; the other derivatives, RBDS-A, and RBDS-B are all hygroscopic; and STL-CPP-Arg2 is slightly hygroscopic.

[0213] Example 15. Investigation of solubility in common solvents or media

[0214] The test was conducted according to the solubility test method in Article 15 (2) of Part IV of the 2020 edition of the Chinese Pharmacopoeia. The solubility of the above-mentioned derivatives in the following solvents was investigated based on the preparation and purification process of each derivative and the commonly used solvents used as carriers.

[0215] Weigh each compound into a fine powder and place it in a certain amount of the following solvent at 25°C ± 2°C. Shake vigorously for 30 seconds every 5 minutes. Observe the dissolution within 30 minutes. If no solute particles are visible, it is considered completely dissolved. The results are shown in Table 7 below:

[0216] Table 7. Solubility test results of various derivatives (25°C) Note 1: “--” The experiment was not carried out.

[0217] In summary, with the exception of STVB-CPP-Arg8, the solubility of the other derivatives in common solvents ranges from 1 mg / ml to 700 mg / ml, meeting the requirements for use as carriers. STVB-CPP-Arg8 is highly soluble in both water and pH 6.8 phosphate buffer.

[0218] Example 16. Stability Study in Simulated Gastrointestinal Fluids Before and After Meals

[0219] (1) Preparation of simulated gastrointestinal fluid before meals:

[0220] Preparation of Fasted State Simulated Gastric Fluid (FaSSGF): Weigh 2 g of sodium chloride, 0.043 g of sodium taurocholate, and 0.015 g of lecithin, add to 800 ml of degassed deionized water, and sonicate at 40°C-50°C to dissolve. Adjust the pH to 1.60 with 1 M HCl, then add 0.1 g of pepsin and fill to 1 L with degassed deionized water. Reconfirm the pH and adjust to 1.20.

[0221] Fasted State Simulated Intestinal Fluid (FaSSIF) FaSSIF preparation: 187 ml FaSSGF was added to 63 ml regulating solution, and the pH was controlled at 6.50.

[0222] Preparation of regulating solution: weigh 8.8056g of maleic acid, 8.8062g of sodium hydroxide, 9.9762g of sodium chloride, 6.618g of sodium taurocholate, and 0.551g of lecithin, add 1000ml of degassed deionized water, and dissolve them by ultrasonication at 40℃-50℃.

[0223] (2) Preparation of postprandial simulated gastrointestinal fluid (without trypsin):

[0224] Preparation of Fed State Simulated Gastric Fluid (FeSSGF): Weigh 13.85 g of sodium chloride, 2.44 g of anhydrous sodium acetate, and 1.028 g of glacial acetic acid, add 1000 ml of degassed deionized water, and stir to dissolve. Then add 1000 ml of whole milk and mix thoroughly. Adjust the pH to 5.00 with concentrated hydrochloric acid.

[0225] Preparation of Fed State Simulated Intestinal Fluid (FeSSIF): Add 100 ml of regulating solution to 150 ml of FeSSGF and control the pH at 5.80.

[0226] Preparation of regulating solution: weigh 15.9654 g of maleic acid, 8.165 g of NaOH, 7.9481 g of NaCl, 13.75 g of sodium taurocholate, 0.6089 g of sodium oleate, and 3.79 g of lecithin, add 1000 ml of degassed deionized water, ultrasonically and stir to dissolve.

[0227] Experiments (1) and (2) selected derivatives of different groups for the experiments. Take appropriate amounts of RBDS-His, STVB-APL, RBSD-Arga and STL-CCP-4Arg, weigh them accurately, add 12 ml of the above-mentioned gastric juice or 20 ml of intestinal juice, and make a solution containing about 0.5 mg of each carrier per 1 ml. Ultrasonicate at 37°C to disperse evenly, place in a 37°C constant temperature air shaker, set the amplitude to 250 rpm, and place it at time 0. Samples were taken according to the table below to detect the concentration of samples at different time points. Sample pretreatment steps for experiments (1) and (2):

[0228] Pre-meal gastric and intestinal fluid samples: 3 ml was collected from each sampling point, centrifuged at 37°C and 15,000 rpm for 5 min, 0.5 ml of the supernatant was added with 1.5 ml of acetonitrile, mixed, filtered through a 0.22 μm nylon filter membrane, 1 ml of the primary filtrate was discarded, and the subsequent filtrate was analyzed by HPLC.

[0229] The HPLC analysis method is as follows:

[0230] An Agilent 1260 high performance liquid chromatograph was used, equipped with a G1311B quaternary high pressure pump, a G1329B autosampler, a G1316A column oven, and a G4212B DAD detector.

[0231] The chromatographic conditions of liquid chromatography are shown in Table 8:

[0232] Table 8. Chromatographic conditions for liquid chromatography

[0233] The concentration of each sampling point was detected by external standard method. The results of each sampling time point are shown in Table 9 below:

[0234] Table 9. Determination results of each derivative in simulated gastric fluid before and after meals at different time points

[0235] Pre-meal simulated gastric fluid contains pepsin. Although the concentrations of each carrier measured during the investigation time were slightly lower than those at time 0, the maximum RSD of the concentration results measured at different time points was 3.1%, and none exceeded 4%. Each carrier had good chemical stability in pre-meal simulated gastric fluid and post-meal simulated gastric fluid. The low pH of pre-meal simulated gastric fluid and pepsin had basically no effect on the stability of the carrier.

[0236] Table 10. Determination results of each derivative in simulated intestinal fluid before and after meal at different time points

[0237] The maximum RSD of the concentrations of each carrier measured in the simulated intestinal fluid (without enzyme) during the investigation time was 2.2%, and none of them exceeded 4%. The chemical stability of each carrier was good in the simulated intestinal fluid before and after a meal.

[0238] Example 17. Analysis of trypsin metabolites

[0239] (1) Solution preparation

[0240] Simulated intestinal fluid containing trypsin: Dissolve 6.8 g of KH2PO4 in 500 ml of water, adjust the pH to 6.8 with 0.4% (w / w) NaOH, add 1 g of trypsin per 100 ml of liquid, mix well, and filter with a 0.2 μm sterile filter for later use.

[0241] Simulated intestinal fluid without trypsin was prepared in the same way.

[0242] (2) Experimental steps

[0243] Measure 30 ml of enzyme-containing simulated intestinal fluid from each experiment, add STVB-CPP-Arg8 to a final concentration of approximately 1.0 mg / ml, sonicate at 20°C to completely dissolve, and immediately incubate in a shaker at 37°C. Samples of 1 ml were collected at 0.5, 1, 1.5, and 2 hours after incubation, except for the 1 hour sampling point. The enzyme reaction was immediately quenched with 1 ml of 0.1 mol / L hydrochloric acid. The mixture was centrifuged at 37°C and 15,000 rpm for 5 minutes. The supernatant (0.5 ml) was added to 1.5 ml of acetonitrile, mixed, and filtered through a 0.22 μm nylon membrane. 1 ml of the initial filtrate was discarded, and the subsequent filtrate was used for qualitative metabolite analysis using LC / MS / MS. A sample of simulated intestinal fluid without enzyme incubated for 1 hour was also collected for comparison.

[0244] Take 10 ml of the sample at the 1 h and 2 h sampling points respectively, immediately quench the enzyme reaction with 10 ml of 0.1 mol / L hydrochloric acid, centrifuge at 37 °C and 15000 rpm for 5 min, take 0.5 ml of the supernatant for analysis, and use the rest of the solution for later use.

[0245] (3) Sample analysis and result interpretation

[0246] The LC / MS / MS analysis method is as follows:

[0247] The high-resolution mass spectrometric data of the present invention were detected using a Waters Xevo TQ-S triple quadrupole mass spectrometer (UPLC / MS / MS), and the ultra-high performance liquid chromatography conditions were as follows:

[0248] The chromatographic column was Agilent ZOBAX SB-C18, 2.1 × 100 mm, 1.8 μm;

[0249] Mobile phase A: 0.1% formic acid-water solution; Mobile phase B: 0.08% formic acid-acetonitrile solution;

[0250] Detection wavelength: 210nm;

[0251] Flow rate: 0.5 mL / min;

[0252] Injection volume: 5 μL;

[0253] Column temperature: 60°C;

[0254] The gradient elution program is as follows:

[0255] Mass spectrometry detection conditions were: capillary voltage: 2.80 kV, drying gas temperature: 350°C, positive / negative ion scanning mode, ESI source, and mass spectrometry scanning range: m / z: 50-1500 Da.

[0256] The analysis and identification results of each metabolite are shown in Table 11 below:

[0257] Table 11. Analysis of major metabolites of STVB-CPP-Arg8 after 1 h of trypsin hydrolysis Note 1: This is the absolute error.

[0258] According to the above results, STVB-CPP-Arg8 was gradually enzymatically hydrolyzed into STVB-CPP-Arg7, STVB-CPP-Arg6, STVB-CPP-Arg5, STVB-CPP-Arg4, STVB-CPP-Arg3, STVB-CPP-Arg2, STVB-CPP-Arg1 and STVB in simulated intestinal fluid containing trypsin at 37°C over time. Metabolites containing different -Arg residues were further enzymatically hydrolyzed into metabolites with fewer -Arg residues. Almost all metabolites could be detected after 1 h of enzymatic hydrolysis. At this time, the area-normalized content of STVB-CPP-Arg8 was about 22.31%, among which the area-normalized content of the metabolite of STVB-CPP-Arg1 was the highest, about 26.76%, followed by STVB-CPP-Arg3, with a normalized content of 22.19%. Compared to the results of the simulated intestinal fluid without trypsin under the same conditions, STVB-CPP-Arg8 was still the main metabolite at 1 hour, and no other metabolites were detected. The liquid phase spectrum is shown in Figure 26.

[0259] Example 18. Investigation of Self-Assembly of Trypsin Metabolites

[0260] The sample solution of Example 17 after enzymatic hydrolysis for 1 h was taken, which contained a normalized content of STVB-CPP-Arg8 of 22.31%, a normalized content of STVB-CPP-Arg1 of 25.76%, a normalized content of STVB-CPP-Arg3 of 22.19%, and the normalized contents of the remaining metabolites ranging from 1.78% to 4.34%. This carrier solution was recorded as STVB-CPP-Arg8-Tps-1h;

[0261] In Example 17, the sample solution after enzymatic hydrolysis for 2 h had normalized contents of major metabolites: STVB-CPP-8: 9.76%, STVB-CPP-Arg5: 11.83%, STVB-CPP-Arg4: 8.78%, STVB-CPP-Arg2: 13.23%, STVB-CPP-Arg1: 17.63%, STVB: 31.52%, and the normalized contents of the remaining metabolites were all below 4%. The carrier solution was recorded as STVB-CPP-Arg8-Tps-2h;

[0262] Curcumin (CCM) was dissolved in a tetrahydrofuran-methanol (2:1, V / V) mixed solvent to form a solution with a curcumin concentration of 0.2 g / g for later use.

[0263] Take 5 ml of the above solution and add curcumin dropwise to 0.3 g while stirring at room temperature. Stop adding and continue stirring for 30 minutes. Recover the organic solvent on a 40°C rotary evaporator until there is no obvious solvent smell. Centrifuge at 13000 rpm for 10 minutes to separate the supernatant. Let it stand at room temperature for 24 hours. If precipitation occurs, centrifuge it again and take the supernatant to detect the concentration, particle size distribution and zeta potential of each sample.

[0264] Measurements were performed using a NanoBrook (model: 90Plus PALS) nanoparticle size analyzer or other instruments with equivalent performance. Dynamic light scattering was used, with room temperature at 25°C and a diffraction angle set at 90°. The measurement time was set to 2 minutes, with three replicates per sample. Deionized water was used as the diluent.

[0265] Zeta potential measurement: The measurement was performed at room temperature of 25°C, the diffraction angle of the Zeta potential measurement was set to 15°C, and 3 replicates were performed for each sample.

[0266] The above solution was centrifuged and allowed to stand for 24 hours without precipitation.

[0267] Table 12. CCM concentration, particle size distribution, PDI, and Zeta potential test results

[0268] According to the above results, using STVB-CPP-Arg8 as a carrier, it enters the small intestine with gastrointestinal peristalsis after oral administration. Under the action of trypsin, even after being enzymatically hydrolyzed into STVB-CPP-Arg7 / Arg6 / Arg5 / Arg4 / Arg3 / Arg2 / Arg1 and STVB, it can still be reassembled.

[0269] Example 19. Determination of critical micelle concentration of each derivative

[0270] Critical micelle concentration (CMC) is an important functional indicator for amphiphilic compounds. Their amphiphilicity enables them to spontaneously associate and assemble into micelles in polar solvents. The critical micelle concentration (CMC) is the minimum concentration at which amphiphilic compound molecules associate to form micelles in polar solvents. Studies have shown that when amphiphilic compounds reach the critical micelle concentration (CMC) in a solvent, many of their physicochemical properties, such as surface tension, solubility, density, viscosity, light scattering intensity, osmotic pressure, and conductivity, undergo sudden changes that deviate from normal behavior. Common methods for determining CMC include surface tension, conductivity, light scattering, dye-based methods, fluorescence spectroscopy, and UV-visible spectroscopy. These methods are based on the relationship between the physicochemical properties of a solution and concentration. The surface tension method can determine the CMC value of anionic and nonionic surfactants. However, if there are highly surface-active polar organic compounds such as long-chain alcohols, higher amines, fatty acids, etc. in the compound, the turning point of the surface tension-concentration curve becomes unclear, making it difficult to determine the critical micelle concentration. At the same time, the determination of surface tension is easily affected by the ambient temperature, the cleanliness of the container, etc. The hydrophobic core of RBDS-A can be regarded as a fatty acid, and therefore, it is not suitable for the surface tension method. The conductivity method is only applicable to ionic surfactants, and the light scattering method has low sensitivity. In combination with the chemical structure of the derivatives, considering the reliability of a single method, the critical micelle concentration of the above derivatives was determined by UV-visible spectrophotometry based on the dye method.

[0271] There are few reports on the CMC of steviol glycosides in the literature. Only Food Chemistry 197(2016)84–91 reported that the CMC of RBDS-A in pH 3.0 and pH 6.7 buffers was 4.5 mM and 5 mM, respectively (determined by fluorescence spectrometry using diphenylhexatriene as a fluorescent probe). Considering that diphenylhexatriene needs to be dissolved in tetrahydrofuran, to reduce solvent interference, iodine / potassium iodide was used as the dye, and water-acetonitrile (V / V, 7:2) was used as the solvent for the CMC determination of these derivatives.

[0272] A commercially available, calibrated 0.05 mol / L iodine titrant (referring to 8006 Iodine Titrant (0.05 mol / L) in the 2020 edition of the Chinese Pharmacopoeia, Part IV) was used as the dye. Based on the CMC value of RBDS-A, solutions of various derivatives were prepared at concentrations near that value. UV-visible full-wavelength scanning revealed strong UV absorption at 286 nm, 346 nm, and 460 nm. The ambient temperature was maintained at 25°C (298 K). RBDS-A was used as a control to verify the feasibility of the method.

[0273] RBDS-A mother solution: Accurately weigh about 100 mg of RBDS-A and place it in a 10 ml volumetric flask. Add water-acetonitrile (V / V, 8:2) to dissolve the solution and dilute to the mark. Shake well and set aside.

[0274] Adopt Agilent Technology's Cary 60 type UV-visible spectrophotometer to carry out absorbance detection. Take several 10ml brown volumetric flasks respectively, measure 20μl 0.05mol / L iodine titrant respectively, add water 1ml, shake up, add RBDS-A mother solution (10mg / ml) 0ml, 1ml, 2ml, 3ml, 4ml, 5ml, 6ml, 7ml and 8ml to each volumetric flask respectively, then add solvent to make up to scale, leave at room temperature for 30min, detect absorbance at 286nm, 346nm and 460nm respectively (n=3). With concentration as horizontal coordinate and absorbance as vertical coordinate, draw two standard curves under the same detection wavelength, and the intersection of the curves is the CMC value. Considering the absorbance measurement accuracy of 460nm iodine, the 286nm and 346nm data are as the criterion.

[0275] Representative compounds with different derivatization groups were selected and their CMCs at wavelengths of 286 nm and 346 nm were determined using the same method.

[0276] Table 13. CMC measurement results of each derivative -298K (25℃)

[0277] According to the above CMC determination results, the CMC value of RBDS-A measured by UV-visible spectrophotometry using iodine as the dye at 286nm and 346nm was approximately 4.0mM, which is basically consistent with the fluorescence detection results in the literature.

[0278] For steviol glycoside derivatives with different structures, the absorbance of iodine-containing micelles constructed with different concentrations of these derivatives was measured using UV spectrophotometry at 286 nm and 346 nm using iodine / potassium iodide as dyes. A regression curve (R ≥ 0.99) was constructed with concentration as the abscissa and absorbance as the ordinate. The intersection of the two curves was the CMC value of each derivative. The results are shown in Table 7. Based on these results, the CMC values ​​of each derivative were lower than the CMC value of RBDS-A measured under the same conditions. Under the same conditions, the concentrations at which these derivatives formed micelles with iodine were lower than the concentration at which RBDS-A assembled into micelles with iodine.

[0279] Example 20. Evaluation of the effects on solubility of delivered components, stability of nanomicelles, and permeability

[0280] The permeability of a compound is determined by its chemical structure but is also influenced by P-gp efflux proteins. The permeability of the derivatives prepared by the present invention when used as delivery vehicles differs from the permeability of the derivatives themselves when assembled into nanocomplexes with the target substance. Therefore, conventional methods for assessing drug permeability often fail to accurately reflect the effect of the derivatives provided by the present invention as delivery vehicles on the permeability of the target substance.

[0281] Taking curcumin (CCM) as an example, its intrinsic solubility in aqueous solution is about 20 μg / ml, which is a poorly soluble substance. Its oil-water partition coefficient LogP is about 4.0, indicating that the molecule is highly lipophilic and should have good permeability. However, according to the literature Eur J Pharm Biopharm. 2011 Feb; 77(2): 275-82. and Mol Nutr Food Res. 2013 Sep; 57(9): 1543-9, curcumin is a low permeability compound, with Papp(A→B) of (2.93±0.94)*10 -6 cm / s, Papp(B→A) is (2.55±0.02)*10 -6 cm / s (Caco-2 monolayer cell model), therefore, curcumin molecules are affected by the efflux protein P-gp, which ultimately makes curcumin molecules low permeability compounds.

[0282] Derivatives with different derivatization groups were selected and nanomicelles were prepared according to the formulation in Table 14 below:

[0283] Table 14. Nanomicelle preparation recipe

[0284] Curcumin was dissolved in a tetrahydrofuran-methanol (2:1, V / V) mixed solvent to form a solution containing curcumin at a concentration of 0.2 g / g and set aside.

[0285] Weigh each amount of carrier in each prescription and dissolve them in water-methanol (8:2, V / V) to prepare a solution containing about 100 mg of carrier per ml. While stirring, slowly add the curcumin solution to the carrier solution. Stop adding when about 0.4 g of curcumin (CCM) is added. Continue stirring for 30 minutes. Recover the organic solvent using a rotary evaporator at 40°C until there is no obvious solvent smell. Centrifuge at 13000 rpm for 10 minutes to separate the supernatant. Let it stand at room temperature for 24 hours. If precipitation occurs, centrifuge it again. Take the supernatant to test the concentration of each sample. At the same time, perform particle size distribution and Zeta-potential detection. The diluent for Zeta potential detection is water. The results are shown in Table 15 below:

[0286] Table 15. Comparative Example 1, F1-F4 concentration, particle size distribution, Zeta potential test results

[0287] Based on the above results, compared with Comparative Example 1, formulations F1-F4 all exhibited significantly higher drug loading, averaging 41-97 times that of Comparative Example 1. Compared to Comparative Example 1, using the same preparation process, formulations F1-F4 exhibited improved particle size distribution uniformity. With the exception of Comparative Example 1, which exhibited essentially no charge, formulations F1-F4 exhibited positive zeta potentials. A 5ml sample of each of the nanomicelle solutions was freeze-dried in a freeze dryer for 48 hours before reconstitution and particle size analysis.

[0288] (1) Solubilization ability

[0289] The intrinsic solubility of curcumin in water is 0.02 mg / ml, while that of comparative example 1 is 0.82 mg / ml. Compared with the intrinsic solubility of curcumin, the solubility of comparative example 1 using RBDS-A as a carrier is 41 times that of curcumin.

[0290] The curcumin nanomicelles constructed from the derivatives of the present invention increase the solubility of curcumin by 1687 to 3961 times the intrinsic solubility of curcumin, achieving an unexpected effect compared with Comparative Example 1.

[0291] (2) Encapsulation efficiency and drug loading

[0292] Encapsulation efficiency = drug concentration in micelles / theoretical drug concentration * 100%

[0293] Drug loading = drug weight in micelles / total weight of micelles * 100%

[0294] The encapsulation rates of the respective formulations were: Comparative Example 1 had an encapsulation rate of 2.25%; and formulations F1 to F4 had encapsulation rates of 80.95%, 90.63%, 97.03% and 99.11%, respectively. The encapsulation rates of formulations F1 to F4 were significantly higher than those of Comparative Example 1.

[0295] The drug loading capacity was calculated based on the curcumin content in the freeze-dried powder of each prescription. The drug loading capacity of Comparative Example 1 was 8.52%; while the drug loading capacity of F1 to F4 ranged from 23.13% to 40.01%. Compared with Comparative Example 1, the drug loading capacity far exceeded that of Comparative Example 1.

[0296] (3) Redissolution after freeze-drying and physical stability of nanomicelles

[0297] An appropriate amount of freeze-dried powder of each prescription was weighed and reconstituted with 5 times, 10 times, 25 times, and 50 times the weight of pH 6.8 phosphate buffer. The reconstitution was examined, and the particle size distribution after 50-fold reconstitution was tested. The results are shown in Table 16 below.

[0298] Table 16. Comparative Example 1 and F1-F4 freeze-dried reconstitution, particle size distribution, and Zeta potential test results

[0299] According to the above results, after freeze-drying, Comparative Example 1 was reconstituted 50 times in pH 6.8 phosphate buffer and still had opalescence. The particle size increased after freeze-drying, and the PDI increased slightly compared to before freeze-drying. The nanoparticles in Comparative Example 1 aggregated during the freeze-drying process, indicating poor physical stability. After freeze-drying, formulations F1 to F4 were reconstituted 50 times in pH 6.8 phosphate buffer and the solutions were all clear. The particle size test results after reconstitution were basically consistent with those before freeze-drying. The PDI did not exceed 0.3, and the particle size distribution was uniform, indicating that the nanomicelles prepared by formulations F1 to F4 had better physical stability.

[0300] (4) Permeability

[0301] Although permeability evaluation methods include the Caco-2 monolayer cell model, in vivo or in situ intestinal perfusion in animals, animal everted intestinal sac method, and isolated intestinal tissue method, in vivo experiments are more convincing than in vitro experiments. Therefore, the nanomicelle solution prepared above was directly gavaged into rats to investigate the relative bioavailability of curcumin in rats. The better the permeability of the curcumin nano oral solution, the more curcumin is absorbed into the blood.

[0302] SPF grade Kunming mice, weighing 18-20g, were used and provided by the China Food and Drug Inspection Institute, license number: SCXK (Beijing) 2022-0002. They were acclimatized to the environment for one week during the day and night alternation before the experiment. All animals were fasted for more than 10 hours overnight before administration, with free drinking water and no water for 1 hour before administration. All animals were randomly divided into 5 groups, 21 mice in each group, 3 mice at each blood sampling point, and the comparative example 1 was made into a suspension (containing CCM 25mg) after the CCM content was converted with lyophilized powder, and 2ml was gavaged. F1-F4 were gavaged with 0.75ml, 0.60ml, 0.42ml and 0.52ml (containing curcumin 25mg) respectively, and no water was given after administration. Free drinking water was given 1h after administration and free food after 2 hours.

[0303] Blood collection time points: 0.3ml-0.5ml of blood was collected before the portal vein enters the liver 2h before administration and 5min, 15min, 30min, 1h, 2h, 3h, and 6h after administration.

[0304] Take 100 μL of plasma, add 600 μL of methanol as a protein precipitant, vortex for 60 seconds, add 84 μL of H2O and 6 μL of internal standard solution, vortex for 30 seconds, let it stand for 10 minutes, centrifuge at 13000 rpm and 4°C for 10 minutes, and take the supernatant for analysis.

[0305] The plasma samples of curcumin were analyzed by liquid chromatography-triple quadrupole mass spectrometry. The chromatographic conditions are shown in Table 17.

[0306] Table 17. Chromatographic conditions

[0307] Since blood was collected before the portal vein entered the liver, the total amount of drug absorbed into the blood through the intestinal mucosa but not metabolized by the liver was measured. Based on the blood drug concentration data measured at different time points, pharmacokinetic parameters were calculated using Phoenix WinNonlin7.0, and AUC was calculated. 0-6h (the total amount of drug absorbed from the intestine into the blood) and its mean and standard deviation. The results are shown in Figure 27.

[0308] The experimental results show that after oral administration to mice in Comparative Example 1, the CCM leakage at the portal vein entrance was measured at 6.55 μg·h / ml after 6 hours. The CCM leakage at the portal vein entrance of mice F1 to F4 after 6 hours was 7.9 times, 9.4 times, 12.8 times, and 14.5 times that of Comparative Example 1, respectively. These were significantly higher than the RBDS-A in Comparative Example 1, demonstrating unexpected results. The RBSD-Argα derivative formulation showed the highest increase in permeability, followed by STL-CPP-Arg2.

[0309] In summary, compared with the existing RBDS-A, the new stevioside derivatives have achieved unexpected results in solubilization, improving nanoparticle stability, drug loading capacity, and improving oral intestinal mucosal permeability.

[0310] Example 21 Metabolic Analysis of STVB-CPP-Arg8 by Human Intestinal Microflora

[0311] (1) Preparation of healthy human fecal homogenate

[0312] Fecal samples were voluntarily provided by three healthy employees within the company, aged 26-32 years, including two males and one female. The above-mentioned donors had not used antibiotics or similar natural sweeteners within 1 month. All fecal samples were taken from fresh morning stool. The fecal samples were weighed and added with 12.5 times the weight volume (W / V) of 0.2M anaerobic potassium phosphate buffer solution (pH 7.0), shaken, and centrifuged at 37°C and 500g for 1 minute. The supernatants of equal volumes of each sample were combined and placed in clean stoppered test tubes. The human fecal homogenate was diluted to 50 times with brain heart infusion broth medium and placed in a 37°C incubator overnight.

[0313] (2) Sample preparation and culture

[0314] In vitro metabolism was performed in sealed, stoppered glass tubes at 37°C under anaerobic conditions. Each sample was prepared in triplicate. STVB was used as a positive control. STVB and STVB-CPP-Arg8 were added at a concentration of 2 mg / ml. The reaction mixture was incubated under anaerobic conditions at 37°C ± 5°C for 0, 8, and 24 hours.

[0315] At the end of each sampling point, each sample was treated with an equal volume of methanol solution containing 5% ammonia to terminate the metabolic reaction, gently vortexed and sonicated for 5 min, centrifuged at 3270 g for 10 min at 4 °C, and the supernatant was transferred to a 1.5 ml centrifuge tube and stored at -80 °C for testing.

[0316] (3) Sample testing

[0317] In vitro bacterial metabolism samples were qualitatively analyzed using LC / MS / MS.

[0318] The LC / MS / MS analysis method is as follows:

[0319] The high-resolution mass spectrometric data of the present invention were detected using a Waters Xevo TQ-S triple quadrupole mass spectrometer (UPLC / MS / MS), and the ultra-high performance liquid chromatography conditions were as follows:

[0320] The chromatographic column was Agilent ZOBAX SB-C18, 2.1 × 100 mm, 1.8 μm;

[0321] Mobile phase A: 0.1% formic acid-water solution; Mobile phase B: 0.08% formic acid-acetonitrile solution;

[0322] Detection wavelength: 210nm;

[0323] Flow rate: 0.5 mL / min;

[0324] Injection volume: 5 μL;

[0325] Column temperature: 60°C;

[0326] The gradient elution program is as follows:

[0327] Mass spectrometry detection conditions were: capillary voltage: 2.80 kV, drying gas temperature: 350°C, positive / negative ion scanning mode, ESI source, and mass spectrometry scanning range: m / z: 50-1500 Da.

[0328] The chromatographic and mass spectrometric conditions for the detection method are as follows:

[0329] The metabolite analysis of the 8-hour control STVB is shown in Table 18:

[0330] Table 18. In vitro metabolic analysis of the control STVB human intestinal flora

[0331] STVB and its metabolites were detected in positive or negative ion mode except (MH) - 、(M+H) + In addition to the mass-to-charge ratio, (M+Na-H) - 、(M+Na+H)+ 、(M+H2O+H) + 、(M+acetonitrile+H) + The mass-to-charge ratio information of the equal-additive peaks was obtained. Based on the total ion current peak area of ​​the mass spectrometer, STL accounted for approximately 72.4% of the 8-hour peak, STVB: 12.8%, and RSBD: 9.1%. These results indicate that the experimental method is reliable.

[0332] The metabolite analysis of STVB-CPP-Arg8 at 8 hours is shown in Table 19:

[0333] Table 19. In vitro metabolic analysis of STVB-CPP-Arg8 human intestinal flora

[0334] The sugar residues in the structure of steviosides are metabolized by β-glucosidase secreted by intestinal flora. The specific metabolic process is as follows:

[0335] Therefore, the above-mentioned human fecal flora is consistent with the above-mentioned metabolic pathway based on the mass spectrometry information of the detected metabolites. After incubation at 37°C in an anaerobic environment for 8 hours, according to the mass spectrometry TIC area normalization results, 40.91% of STVB-CPP-Arg8 was not metabolized, the detection amount of RSBD-CPP-Arg8 was 37.61%, and the detection amount of STL-CPP-Arg8 was 21.93%. Compared with the control, the metabolic rate slowed down significantly, probably because the steric hindrance increased after the connection of octaarginine, and therefore the degradation rate also decreased. From the perspective of drug delivery, this can prolong the residence time of the delivered drug in the intestine, thereby better exerting its efficacy. For details of the 8-hour metabolic spectrum, please see Figures 28-1 / 28-2.

[0336] The specific embodiments described above further illustrate the objectives, technical solutions and beneficial effects of the present invention in detail. It should be understood that the above are only specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A delivery vector, characterized in that The delivery carrier is a compound represented by the general formula Ia, or a salt thereof, or a hydrate thereof, or any mixture of the compound represented by the general formula Ia, a salt thereof, and a hydrate thereof, In formula Ia, R1 and R2 are independent of each other, R1 is selected from hydroxyl or glycosyl, and R2 is selected from wherein R3 is selected from alkali metals, alkaline earth metals, H, C1-C10 straight chain, branched chain, cyclic alkanes or alkanes with hydroxyl, amino, thiol substituted side chains, polymer groups or sugar groups; R4 and R5 are independently selected from H, C1-C 10 The invention relates to a straight chain, branched chain, or cyclic alkane or an alkane whose side chain is substituted with a hydroxyl group, an amino group, or a thiol group, a polymer group, an amino acid or an amino acid residue, an aminoamide, or a short peptide condensed with any number of 1 to 9 amino acids.

2. The delivery vector according to claim 1, characterized in that The sugar group is a pyranose monosaccharide or oligosaccharide.

3. The delivery vehicle according to claim 1, characterized in that The sugar group is selected from pyranose glucose, galactopyranose, pyranose mannosyl, pyranose deoxyglucose, pyranose rhamnose, pyranose xylose, pyranose glucosamine, and oligosaccharides formed by polymerization of any number of 1 to 9 pyranose monosaccharides at any position.

4. The delivery vector according to any one of claims 1 to 3, characterized in that The absolute configuration of the sugar group is D-type or L-type, and the relative configuration of the terminal carbon is α-type or β-type.

5. The delivery vector according to claim 1, characterized in that The alkali metal is selected from sodium, potassium or lithium.

6. The delivery vector according to claim 1, wherein the polymer group is selected from polyethylene glycol, polydopamine, polyarginine amide or polyglutamic acid.

7. The delivery vector according to claim 1, characterized in that The amino acid or amino acid residue is selected from alanine, arginine, aspartic acid, cysteine, glutamic acid, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, tyrosine or valine, and the aminoamide is selected from the aminoamide corresponding to the amino acid.

8. The delivery vector according to claim 1, characterized in that The short peptide is a cell-penetrating-peptide (CPP).

9. The delivery vector according to claim 8, characterized in that The cell-penetrating peptide is octaarginine.

10. The delivery vector according to claim 1, characterized in that The R1 is selected from hydroxyl, 11. The delivery vehicle according to claim 1, characterized in that The R2 is selected from Wherein n is 0-9, X=OH and / or NH2.

12. The delivery vector according to claim 1, characterized in that The optical purity of the compound represented by the general formula Ia is not limited, and it can be in R and / or S configuration, with any enantiomeric excess value or any diastereomeric excess value.

13. A delivery carrier, which is at least one compound selected from the group consisting of the following, or a salt thereof, or a hydrate thereof, or an in vivo metabolite thereof: (1) (1R,4S,5R,9S,10R,13S)-13-[(2S,3R,4S,5R,6R)-5-hydroxy-6-(hydroxymethyl)-3,4-bis[[(2S,3R),4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl]oxy]oxa-2-yl]oxy-5,9-dimethyl-14-methylenetetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylate sodium (RBDS-B-Na), (2) (4R, 4aS, 6aR, 9S, 11aR, 11bS)-N-((S)-1-amino-3-(1H-imidazol-4-yl)-1-oxopropan-2-yl)-9-((2S, 3R, 4S, 5R, 6R)-5-hydroxy-6-(hydroxymethyl)-3,4-bis(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide (RBDS-B-His), (3) (4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-((2S, 3R, 4S, 5R, 6R)-5-hydroxy-6-(hydroxymethyl)-3,4-bis((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2- 1-((S)-2-hydroxypropyl)-4,11-dimethyl-8-methylenetetradecahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide (RBDS-B-APL), (4) (1R,4S,5R,9S,10R,13S)-13-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl]oxyoxa-2-yl]oxy-5,9-dimethyl-14-methylenetetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylic acid (STVB), (5) (6S, 9S, 12S, 15S, 18S, 21S, 24S, 27S)-1-amino-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-(((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro ... Hydrogen-1H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetradecahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-9,12,15,18,21,24,27-hepta(3-guanidinopropyl)-1-imino-7,10,16,19,22,5-heptyloxy-2,8,11,14,17,20,23,26-octaazaoctadecane-28oleic acid (STVB-CPP-Arg8), (6) (4R, 4aS, 6aR, 9S, 11aR, 11bS)-N-((S)-1-amino-3-(1H-imidazol-4-yl)-1-oxopropan-2-yl)-9-((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide (STVB-His), (7) (4R, 4aS, 6aR, 9S, 11aR, 11bS)-N-((S)-1-amino-5-((diaminoethylene)amino)-1-oxopentan-2-yl)-9-((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide (STVB-Arga), (8) (4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-N-((S)-2-hydroxypropyl)-4,11-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide (STVB-APL), (9) (4R, 4aS, 6aR, 9S, 11aR, 11bS)-N-((S)-1-amino-5-((diaminoethylene)amino)-1-oxopentan-2-yl)-4,11-dimethyl-8-methylene-9-((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetradecahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide (RBSD-Arga), (10) (6S, 9S, 12S, 15S, 18S, 21S, 24S, 27S)-1-amino-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-4,11b-dimethyl-8-methylene-9-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetradecahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide) -9,12,15,18,21,24,27-hepta(3-guanidinopropyl)-1-imino-7,10,13,16,19,22,25-heptyloxy-2,8,11,14,17,20,23,26-octaazaoctadecane-28-carboxylic acid (RBSD-CPP-Arg8), (11) (6S, 9S, 12S, 15S, 18S, 21S, 24S, 27S)-1-amino-9,12,15,18,21,24,27-heptyl(3-guanidinopropyl)-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-hydroxy-4,11-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-amide)-1-imino-7,10,13,16,19,22,25-heptyloxy-2,8,11,14,17,20,23,26-octaazaoctacosanoic acid (STL-CPP-Arg8), (12) (6S, 9S, 12S, 15S)-1-amino-9,12,15-tris(3-guanidinopropyl)-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-hydroxy-4,11-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-1-imino-7,10,13-trioxy-2,8,11,14-tetraazahexadecane-16-carboxylic acid (STL-CPP-Arg4), (13) (S)-5-guanidine-2-((S)-5-guanidine-2-(4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-hydroxy-4,11-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-amido)pentanamido)pentanoic acid (STL-CPP-Arg2), (14) (6S, 9S, 12S, 15S, 18S, 21S, 24S)-1-amino-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-(((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl) oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-9,12,15,18,21,24-hexa(3-guanidinopropyl)-1-imino-7,10,13,16,19,22-hexaoxo-2,8,11,14,17,20,23-heptaazapentacan-25 acid (STVB-CPP-Arg7), (15) (6S, 9S, 12S, 15S, 18S, 21S)-1-amino-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-(((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)- )oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-9,12,15,18,21-penta(3-guanidinepropyl)-1-imino-7,10,13,16,19-pentaoxo-2,8,11,14,17,20-hexaazadocosa-22-oic acid (STVB-CPP-Arg6), (16) (6S, 9S, 12S, 15S, 18S)-1-amino-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-(((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran- 2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-9,12,15,18-tetrakis(3-guanidinopropyl)-1-imino-7,10,13,16-tetraoxo-2,8,11,14,17-pentaazanonadecan-19-oic acid (STVB-CPP-Arg5), (17) (6S, 9S, 12S, 15S)-1-amino-6-((4R, 4aS, 6aR, 9S, 11aR, 11bS)-9-(((2S, 3R, 4S, 5S, 6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S, 3R, 4S, 5S, 6R)-3,4,5-trihydroxy- Hydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-9,12,15-tris(3-guanidinepropyl)-1-imino-7,10,13-trioxy-2,8,11,14-tetraazahexadecane-16-oic acid (STVB-CPP-Arg4), (18) (S)-2-((S)-2-(S)-2-[(4R,4aS,6aR,9S,11aR,11bS)-9-((2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-1H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-5-guanidine amide)-5-guanidine amide)-5-guanidine amide)-5-guanidine amide (STVB-CPP-Arg3), (19) (S)-2-((S)-2-((4R,4aS,6aR,9S,11aR,11bS)-9-((2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-1H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetradecahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-5-guanidine amide)-5-guanidine valeric acid (STVB-CPP-Arg2), and (20) (S)-2-((4R,4aS,6aR,9S,11aR,11bS)-9-((2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)tetrahydro-1H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide)-5-guanidine pentanoic acid (STVB-CPP-Arg).

14. A method for preparing the delivery vector of claim 1, characterized in that The steps include: (1) Step 1: subjecting compound 1 to a hydrolysis reaction to obtain compound 2, wherein when compound 1 is in the form of a carboxylic acid, step 2 is directly performed; (2) Step 2: Protect the R1 group in compound 2 with a protecting group P to obtain compound 3; (3) Step 3: subjecting compound 3 to a condensation reaction to obtain compound 4; (4) Step 4: subjecting compound 4 to a deprotection reaction to obtain a compound of formula Ia, Wherein R1 in compound 1, compound 2, compound 3 and R2 in compound 4 are as defined in any one of claims 1-12.

15. The preparation method according to claim 14, characterized in that The hydrolysis reaction is carried out in a solvent (preferably water) under alkaline conditions (preferably sodium hydroxide or potassium hydroxide), wherein compound 1 is selected from Stevioside (STVS), Rebaudioside A (RBDS-A), Dulcoside A (DCS-A), Rubusoside (RBSD), Rubusoside acid (RBSD-Acid), Steviolbioside (STVB), Rebaudioside B (RBDS-B), Rebaudioside C (RBDS-C) acid (RBDS-C-Acid), Steviol (STL) or Isosteviol, or characterized in that, when compound 1 is Steviolbioside (STVB), Rebaudioside B (RBDS-B), Rebaudioside When the acid form of C (RBDS-C) (RBDS-C-Acid), steviol or rubusoside (RBSD) (RBSD-Acid) is used, the preparation method does not require step 1.

16. The preparation method according to claim 14 or 15, characterized in that In step 2, the protecting group is a protecting group of sugar or hydroxyl, preferably a silyl group or an acetyl group, preferably the sugar or hydroxyl group is fully acetylated, for example, the protected compound 3 is selected from RBDS-B-Ac, STVB-Ac, RBSD-Acid-Ac, DCS-A-Acid-Ac or STL-Ac, and the structural formulas are shown below, Wherein: Ac represents acetyl.

17. The preparation method according to any one of claims 14 to 16, characterized in that The condensation reaction of step 3 is that compound 2 reacts with a sugar, amine or alcohol having an R2 group to form a glycoside, amide or ester. Preferably, the carboxyl group of compound 2 is activated with oxalyl chloride to form an acyl chloride, and then reacts with an amino compound in the presence of alkaline conditions (such as triethylamine).

18. The preparation method according to any one of claims 14 to 17, wherein when the protecting group P is an acetyl group, the step 4 is to use an alcohol solvent (such as methanol or ethanol) and carry out a deprotection reaction under a strong base condition (such as potassium tert-butoxide or sodium methoxide or sodium ethoxide).

19. A delivery kit, characterized in that Comprising the delivery vehicle according to any one of claims 1 to 13 and an active ingredient.

20. The delivery kit according to claim 19, characterized in that The active ingredient is a biologically active agent, a chemically active agent or an adjuvant.

21. The delivery kit according to claim 19 or 20, characterized in that The active ingredient is selected from: proteins / polypeptides, polysaccharides / oligosaccharides, nucleic acids or nucleic acid fragments, nanocarriers (such as liposomes, polymer micelles, inorganic nanoparticles), lipids, nutrients, organic small molecule compounds, phage particles, superparamagnetic substances, vaccines, cells, or any combination thereof.

22. The delivery kit according to any one of claims 19 to 21, characterized in that The weight ratio of the active ingredient to the delivery vehicle is about 1:0.5-1:150 (preferably 1:1, 1:10, 1:20, 1:30, 1:40, 1:50, 1:60, 1:70, 1:80, 1:90, 1:100, 1:110, 1:120, 1:130 or 1:140).

23. The delivery kit according to any one of claims 19 to 22, wherein one of the active ingredient and the delivery system is in a liquid state or a solid state, or the active ingredient and the delivery system are both in a liquid state or a solid state.

24. The delivery kit according to any one of claims 19 to 23, characterized in that The active ingredient and the delivery vehicle exist independently, or the active ingredient and the delivery vehicle exist in a mixture.

25. The delivery kit according to any one of claims 19 to 24, characterized in that The active ingredient and the delivery vehicle are in a form suitable for oral, sublingual, buccal, intraduodenal, intracolonic, rectal, vaginal, mucosal, subcutaneous, intramuscular, intravenous, arterial, cutaneous, pulmonary, intranasal, aural, or ocular administration.

26. The delivery kit according to any one of claims 19 to 25, characterized in that The active ingredient and / or the delivery vehicle is in the form of micelles, liposomes, nanoparticles, microspheres, microcapsules, solid dispersions, molecular compositions or hydrogels.

27. The delivery kit according to any one of claims 19 to 26, characterized in that When the active ingredient and the delivery vehicle exist independently, the two are assembled after being mixed and dissolved during use; when the active ingredient and the delivery vehicle exist in a mixed state, the active ingredient and the delivery vehicle in a solid state are assembled after being dissolved.

28. The delivery kit according to any one of claims 19 to 27, characterized in that The assembly of the active ingredient and the delivery carrier in the delivery system is in situ assembly.

29. The delivery kit according to claim 28, characterized in that The in situ assembly is self-assembly in the gastrointestinal tract, local self-assembly in the mucosa, self-assembly in the blood or self-assembly in the target cells.

30. The delivery kit according to any one of claims 28-29, characterized in that The in situ assembly is the in situ assembly of the delivery vector according to any one of claims 1 to 13 with the metabolites after enzyme or microbial metabolism and the active ingredient.

31. The delivery kit according to any one of claims 19 to 25, characterized in that The active ingredient and / or the delivery vehicle is in a dosage form selected from the following: tablets, capsules, oral liquids, drops, gels, granules, emulsions, creams, injections, eye drops, inhalants, sprays, aerosols or patches.

32. The delivery kit according to any one of claims 19 to 31, characterized in that The delivery system further comprises a polymer, preferably, the polymer has a group that can be dissociated under physiological conditions, and / or has more than 10 groups that can provide hydrogen donors or hydrogen acceptors, more preferably, the polymer is selected from the group consisting of: (1) Cellulose polymers, such as hydroxypropyl methylcellulose (HPMC), low-substituted hydroxypropyl cellulose (L-HPC), hydroxypropyl methylcellulose acetate succinate (HPMCAS), sodium carboxymethyl cellulose (CMC-Na) or hydroxyethyl cellulose; (2) Synthetic polymers: such as polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyvinyl pyrrolidone (PVP), copolyvidone (PVP-VA64), polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol graft copolymer (Soluplus), polyglutamic acid (PGA), polydopamine (PDA), polylactic acid (PLA), poly-L-lactic acid (PLLA), polylactic acid-glycolic acid copolymer (PLGA), sodium polyacrylate, siRNA and its derivatives (alkyl chains to improve lipophilicity), mRNA and its derivatives (alkyl chains to improve lipophilicity) or antisense oligonucleotides and their derivatives (alkyl chains to improve lipophilicity); (3) polysaccharides and their derivatives (e.g., carboxymethylated, sulfonylated, phosphorylated, acylated or hydroxypropylated, cationic, anionic or other derivatives), such as carrageenan, guar gum, gum arabic, locust bean gum, konjac gum, agar, gelatin, pectin, gellan gum, hyaluronic acid (HA), aminodextran, chondroitin sulfate, dermatan sulfate, heparin, keratan sulfate, heparan sulfate, sodium alginate, propylene glycol alginate, agar, fucoidan, cyclodextrin and its derivatives, chitosan and its derivatives (e.g., acylated, carboxylated, alkylated and quaternized, etc.), soy protein, vegetable protein or bovine serum albumin; and (4) Any combination of (1)-(3).

33. The delivery kit according to any one of claims 19 to 32, characterized in that The delivery system further comprises an additive selected from excipients, diluents, binders, disintegrants, lubricants, flavoring agents, pH regulators, osmotic pressure regulators, thickeners, plasticizers, colorants, film formers, preservatives or solvents, and any combination thereof.

34. The delivery kit according to any one of claims 19 to 33, for use selected from the group consisting of: (1) For delivery of active ingredients to mammals (such as rodents, cattle, pigs, dogs, cats, primates, humans) or birds (such as chickens, ducks, geese); (2) Use as solubilizer in agricultural chemistry, in particular in formulations containing pesticides, herbicides, fungicides or insecticides, in particular as crop protection agents for spray, pour or immersion mixtures; (3) Use as a preservative or flavoring agent for meat, vegetables, fruits, food, or beverages; and (4) Used in cosmetics and fine chemical products to increase solubility, improve stability, and promote penetration.

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