Kit for forming supramolecular self-assembly complex and use thereof

Abstract

The present invention relates to a kit for forming a supramolecular self-assembly complex and a use thereof. The kit comprises: a package 1, a package 2, and a package 3; the package 1 comprises one or more carriers I providing a positive charge or a negative charge under physiological conditions; the package 2 comprises one or more polymers having dissociable groups under physiological conditions and / or 10 groups or more capable of providing a hydrogen donor and / or a hydrogen acceptor; and the package 3 comprises one or more carriers II formed by covalently linking a unit A and a unit B, wherein the unit A is a hydrophilic moiety, and the unit B is a hydrophobic moiety.

Description

Kit for forming supramolecular self-assembly complex and its application Technical Field

[0001] The present invention belongs to the field of life sciences and specifically relates to an in situ supramolecular self-assembly nano-delivery system and its preparation. This in situ supramolecular self-assembly nano-delivery system can achieve precise delivery of functional components and realize precise treatment or diagnosis. Background Art

[0002] With the rapid development of nanotechnology, nanomedicine, particularly nanomedicine, nanocarriers, and precision drug delivery, has achieved significant breakthroughs. Nanocarrier-based nanodelivery systems, such as liposomes, solid lipid nanoparticles, lipid-based nanoparticles, polymer nanoparticles, dendrimer nanocarriers, and inorganic nanocarrier delivery, are widely reported in literature and patents. Among them, liposomes, polymer nanoparticles, dendrimer nanocarriers, and lipid nanoparticles have already been translated into clinical products. However, because these nanodrug delivery systems cannot withstand the complex gastrointestinal environment that damages the delivery system's integrity or effectively cross the gastrointestinal barrier, currently marketed products are primarily administered via injection.

[0003] The advantages of the in situ nano-delivery system with adaptive properties provided by the present invention are as follows:

[0004] (1) Compared with the existing technology, the drug loading capacity is greatly improved;

[0005] (2) By introducing adaptive carriers, the penetration efficiency of target delivery substances throughout the gastrointestinal tract is greatly improved;

[0006] (3) Glycoside carrier conjugates significantly increase the water solubility of the carrier, allowing some drugs whose absorption is limited by dissolution rate to be developed into fast-acting emergency drugs;

[0007] (4) The interaction strength between glycoside carriers and target delivery substances is improved, so that the constructed supramolecular self-assembled nanoparticles have added strong electrostatic connections on the basis of the original loose connections, improving the integrity of self-assembled nanoparticles in complex delivery scenarios, and successfully solving the problems of drug leakage in lipid nano-delivery and sudden drug release caused by molecular accumulation during production or storage;

[0008] (5) Changing the effects of the physical and chemical properties of drug structure on drug absorption, transport, metabolism, and elimination, thereby better exerting the efficacy of drugs and reducing toxic side effects;

[0009] (6) The supramolecular self-assembly nanocomposition provided by the present invention solves the clinical limitation that lipid nano-delivery and solid lipid nano-delivery cannot withstand the damage or metabolism of the complex gastrointestinal environment to the delivery system and must be administered by injection;

[0010] (7) Higher drug loading capacity and better physicochemical stability;

[0011] (8) By assembling in situ in vivo, the absorption window of the drug during gastrointestinal transit is expanded, the drug delivery efficiency and onset time are improved, and a more reliable long-term release and long-term absorption is provided;

[0012] (9) Improved patients’ medication compliance, medication safety, and effectiveness;

[0013] (10) By improving drug delivery efficiency, the clinical drug dosage of patients can be greatly reduced, thereby significantly reducing the toxic and side effects related to drug dosage;

[0014] (11) The preparation process is simple, reliable, and easy to industrialize.

[0015] Summary of the Invention

[0016] The inventors discovered that structural derivatization of stevioside carriers significantly improved the solubility of such carriers under physiological pH conditions. These carriers were assembled with other carriers or high molecular weight polymers into supramolecular self-assembled complexes, resulting in a significant increase in the apparent permeation rate across the gastrointestinal mucosa and a higher oral bioavailability in animals.

[0017] In addition, the inventors also found that the combined use of glycoside molecules with different chemical structures can greatly increase the drug loading capacity and achieve higher cross-biological barrier transport efficiency compared with the use of a single carrier or a single carrier in combination with a polymer.

[0018] What surprised the inventors even more was that the supramolecular self-assembly complex constructed with the participation of stevioside conjugates can be absorbed into the blood through intestinal mucosal cells, and can also enter the colon with the peristalsis of the digestive tract. After enzyme metabolism, it will undergo in situ self-assembly, so that the assembled complex has a negative charge, making it difficult to be quickly cleared or more conducive to transportation and absorption in the colon.

[0019] Specifically, the present invention relates to the following aspects:

[0020] 1. A kit for forming a supramolecular self-assembly complex, comprising: package 1, package 2 and package 3, wherein

[0021] Package 1 contains one or more carriers I, which provide positive or negative charges under physiological conditions, and the carrier I is as shown in structural formula (1-1) or (1-2),

[0022] Wherein R1 in the structural formula (1-1) / (1-2) is selected from the following groups:

[0023] Wherein R2 in the structural formula (1-1) / (1-2) is selected from the following groups:

[0024] Package 2 comprises one or more polymers having groups that can dissociate under physiological conditions and / or having more than 10 groups that can provide hydrogen donors and / or hydrogen acceptors, and

[0025] Package 3 contains one or more carriers II, wherein the carrier II is composed of a covalently linked unit A and a unit B, wherein unit A is a hydrophilic portion and unit B is a hydrophobic portion, wherein unit A is located on one side or both sides of unit B. When unit A is located on both sides of unit B, the units A may be the same or different; wherein the number ratio of unit A to unit B is (0:1)-(7:1),

[0026] Wherein unit B is selected from one or more of B1, or one or more of B2, wherein:

[0027] (1) B1, molecular formula is (C5H8) n The compound represented by, and its alcohol, aldehyde, ketone, carboxylic acid or ester derivatives, wherein n=4 or 6;

[0028] (2) B2, a compound composed of two benzene rings (A-ring and B-ring) connected to each other by a central three-carbon chain, with 2-phenylchromone as the basic mother nucleus and a C6-C3-C6 basic skeleton, and its derivatives (such as halides or amino acid esters, the halides are such as chlorine or fluorine, and the amino acid esters are such as esters formed by the hydroxyl group on C6 and amino acids);

[0029] The unit A is selected from one or more of A1 or A1 derivatives, wherein:

[0030] A1 is selected from D-type or L-type monosaccharides, disaccharides, trisaccharides or a combination thereof,

[0031] wherein the monosaccharide is selected from the group consisting of glucose, rhamnose, galactose, arabinose, xylose, mannose, fucose, apiose, glucuronic acid, glucosamine, galacturonic acid, acetylamino sugars and combinations thereof, and the relative configuration of each glycosyl terminal carbon can be α-type or β-type;

[0032] Disaccharide selected from the group consisting of sophorose (glc1-2glc), gentiobiose (glC1-6glc), rutinose (rha1-6glc), neohesperidose (rha1-6glC), acaciabiose (rha1-6gal) and combinations thereof;

[0033] trisaccharide selected from the group consisting of gentiotriose, sophorotriose (glc1-2glc1-2glc) and combinations thereof;

[0034] The derivative of A1 is an acylated form of A1, such as 2-acetylglucose and caffeoylglucose.

[0035] 2. The kit according to item 1, wherein the amino acids in the carrier I are selected from alanine, arginine, aspartic acid, cysteine, glutamic acid, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, tyrosine, valine, or their corresponding aminoamides (e.g., arginineamide, histidineamide, homoarginine), and polyamino acids thereof, preferably L-configured amino acids,

[0036] Alternatively, the number of amino acids in the cell-penetrating peptide in the vector I is 20 or less.

[0037] 3. The kit according to item 1, wherein the carrier I is represented by structural formula (1-1).

[0038] 4. The kit according to item 1, wherein the ester derivative of B1 is an ester formed from a carboxylic acid derivative of B1, such as gluconate ester, polyethylene glycol ester, propylene glycol ester, methyl ester, ethyl ester or 1-aminoglycine ester.

[0039] 5. The kit according to item 1, wherein the polymer in the package 2 is selected from the group consisting of:

[0040] (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;

[0041] (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);

[0042] (3) Polysaccharides and their derivatives (such as 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 (such as acylated, carboxylated, alkylated and quaternized), soy protein, vegetable protein or bovine serum albumin;

[0043] and combinations thereof.

[0044] 6. The kit according to any one of items 1 to 5, characterized in that the hydrophobic unit B is a tetracyclic diterpene, a tetracyclic triterpene, a pentacyclic triterpene, or an alcohol, aldehyde, ketone, carboxylic acid or ester derivative thereof.

[0045] 7. The kit according to any one of items 1 to 6, characterized in that the hydrophobic unit B is a kaurene-type tetracyclic diterpene and its derivatives (such as alcohols, aldehydes, ketones, carboxylic acids or esters).

[0046] 8. The kit according to any one of items 1 to 7, characterized in that the hydrophobic unit B is one or more of dammarane type, gansuline type, cycloartane type, lanolin type, cucurbitane type, azadirachtin type and prototerpane type, and their derivatives (such as alcohols and esters), more preferably cucurbitane type and its derivatives (such as alcohols and esters).

[0047] 9. The kit according to any one of items 1 to 8, characterized in that the hydrophobic unit B is one or more of oleanane type, ursane type, lupeane type, suberane type, hobeane type and isohobeane type, and their derivatives (such as alcohols, ketones, carboxylic acids and esters).

[0048] 10. The kit according to any one of items 1 to 9, characterized in that when (1) unit B1 is a kaurene-type tetracyclic diterpene or a pharmaceutically acceptable salt thereof (such as a sodium salt, potassium salt, ammonium salt), or a hydrate of a kaurene-type tetracyclic diterpene or a pharmaceutically acceptable salt thereof (such as a hydrate formed with 0.5, 1, 1.5, 2, 2.5 or 3 crystal waters), the carrier II has a structure of structural formula (2);

[0049] Wherein R1 and R2 represent unit A, and R1 and R2 exist simultaneously, and R1, R2 are independently selected from the following groups:

[0050] or

[0051] (2) When unit B1 is a cucurbitane-type tetracyclic triterpene or a pharmaceutically acceptable hydrate or solvate thereof, the carrier II has the structural formula (3):

[0052] Wherein R1 and R2 represent unit A, and R1 and R2 exist simultaneously or alone, and R1, R2R3, and R4 are independently selected from the following groups:

[0053] 11. The kit according to any one of items 1 to 10, characterized in that the unit B is B2 and is selected from the group consisting of flavonols (as represented by structural formula (4-1)), or dihydrochalcone (as represented by structural formula (4-2)), and a combination thereof, wherein the structure of structural formula (4-1) or structural formula (4-2) is as follows:

[0054] 12. The kit according to any one of items 1 to 11, wherein the carrier I is selected from the group consisting of:

[0055] (1) (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),

[0056] (2) (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),

[0057] (3) (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),

[0058] (4) (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),

[0059] (5) (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),

[0060] (6) (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-carboxylate sodium (STVB-Na),

[0061] Any combination of the compounds shown in (7) and (1)-(6).

[0062] 13. The kit according to any one of items 1 to 12, wherein the carrier II is selected from the group consisting of the following compounds:

[0063] (1) [(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](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 (RBDS-M),

[0064] (2) (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](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 (RBDS-D),

[0065] (3) [(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl](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 (RBDS-A),

[0066] (4) (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-carboxylic acid (RBDS-B),

[0067] (5) [(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl](1R,4S,5R,9S,10R,13S)-13-[(2S,3R,4S,5R,6R)-5-hydroxy-6-(hydroxymethyl)-4-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl]oxy-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxa-2-yl]oxyoxa-2-yl]oxy-5,9-dimethyl-14-methylenetetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylate (RBDS-C),

[0068] (6) [(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl](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-carboxylate (STVS),

[0069] (7) [(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl](1R,4S,5R,9S,10R,13S)-13-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4R),5R,6S)-3,4,5-trihydroxy-6-methyloxa-2-yl]oxyoxa-2-yl]oxy-5,9-dimethyl-14-methylenetetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylate (DCS-A),

[0070] (8) (4R,4aS,6aR,9S,11aR,11bS)-9-(((2S,3R,4S,5R,6R)-5-hydroxy-6-(hydroxymethyl)-4-(((2S,3R,4S),5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-3-((((2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxylic acid (RBDS-C-Acid),

[0071] (9) (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),

[0072] (10) [(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl](1R,4S,5R,9S,10R,13S)-5,9-dimethyl-14-methylene-13-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl]oxytetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylate (RBSD),

[0073] (11)(2R,3S,4S,5R,6R)-2-(hydroxymethyl)-6-({[(2R,3S,4S,5R,6S)-3,4-dihydroxy-6-{[(3R,6R)-2-hydroxy-6-[(1R,3aS,3bS,7S,9aR,9bR,10R,11aR)-10-hydroxy-7-{[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-({[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)-3,4,5,6-tetrahydro-2H-pyran-2-yl]oxy}methyl)-3,4,5, 6-tetrahydro-2H-pyran-2-yl]oxy}-3a,6,6,9b,11a-pentamethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetrahydro-1H-cyclopenta[1,2-a]phenanthren-1-yl]-2-methylhept-3-yl]oxy}-5-{[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)-3,4,5,6-tetrahydro-2H-pyran-2-yl]oxy}-3,4,5,6-tetrahydro-2H-pyran-2-yl]methyl}oxy)tetrahydropyran-3,4,5-triol (MGSD-V),

[0074] (12)(3S,8S,9R,10R,13R,14S,17R)-17-((2R,5R)-5-((((2S,3R,4S,5S,6R)-4,5-dihydroxy-3-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)methyl)tetrahydro-2H-pyran-2-yl)oxy) -6-hydroxy-6-methylheptan-2-yl)-4,4,9,13,14-pentamethyl-3-(((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-((((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)-1,2,3,4,7,8,9,10,12,13,14,15,16,17-tetradecahydro-11H-cyclopenta[a]phenanthrene-11-one (OMGSD-V),

[0075] (13)(2S,3R,4S,5S,6R)-2-(((2S,3R,4S,5S,6R)-4,5-dihydroxy-2-(((3R,6R)-2-hydroxy-6-((3S,8S,9R,10R,11R,13R,14S,17R)-11-hydroxy-4,4,9,13,14-pentamethyl-3-(((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)- 2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthren-17-yl)-2-methylheptan-3-yl)oxy)-6-((((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)methyl)tetrahydro-2H-pyran-3-yl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (SMSD-I),

[0076] (14) (2R,3R,4S,5S,6R)-2-(((3S,8R,9R,10S,11R,13R,14S,17R)-17-((2R,5R)-5,6-dihydroxy-6-methylheptan-2-yl)-11-hydroxy-4,4,13,14-tetramethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthren-3-yl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (MGSD-IE1),

[0077] (15) (2S)-7-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-2-yl]oxy-5-hydroxy-2-(3-hydroxy-4-methoxyphenyl)-2,3-dihydrochromen-4-one (NHPD),

[0078] (16) 1-[4-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxy-2-yl]oxyoxan-2-yl]oxy-2,6-dihydroxyphenyl]-3-(3-hydroxy-4-methoxyphenyl)propan-1-one (NHDC),

[0079] (17) 1-[4-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-2-yl]oxy-2,6-dihydroxyphenyl]-3-(4-hydroxyphenyl)propan-1-one (NRGDC),

[0080] (18) methyl (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)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxylate (RBDS-B-Methyl),

[0081] (19) Ethyl (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)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxylate (RBDS-B-Ethyl),

[0082] (20) Conjugates of steviol glycosides (stevioside, RBDS-B, STVB, RBSD)-HA, and

[0083] (21) Any combination of the compounds shown in (1)-(20).

[0084] 14. The kit according to any one of items 1 to 13, wherein the kit further comprises a package 4, wherein the package 4 comprises an active ingredient, wherein the active ingredient is a compound, preferably a drug, a monomeric component or component extract of a natural product, a nutrient, a contrast agent and / or a diagnostic agent.

[0085] 15. The kit according to item 14, wherein the mass ratio of the active ingredient to the sum of the carrier I and the carrier II is 1:0.5 to 1:50 (preferably 1:1, 1:5, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40 or 1:45), the carrier I accounts for 0% to 75% (preferably 0% to 70%, 0% to 65%, 0% to 60%, 0% to 55%, 0% to 50%, 0-40%, 0-30%, 0-20% or 0-10%) of the total mass of the carrier I and the carrier II; and the mass ratio of the active ingredient to the polymer is 1:0 to 1:25 (preferably 1:0 to 1:20, 1:0 to 1:15, 1:0 to 1:10, 1:0 to 1:5).

[0086] 16. The kit according to item 14, wherein the drug is selected from small molecule compound drugs,

[0087] The nutrient is a vitamin or coenzyme Q10, preferably a vitamin selected from the group consisting of one or more of vitamin A, B vitamins, vitamin E, vitamin K, and vitamin D;

[0088] The contrast agent is selected from tumor fluorescent agents, CT tracers (such as iodine nanocontrast agents, gold nanocontrast agents, tantalum oxide nanocontrast agents, bismuth nanocontrast agents or lanthanide nanocontrast agents), MRI tracers (such as paramagnetic contrast agents, ferromagnetic contrast agents or supermagnetic contrast agents) and radionuclide tracers.

[0089] 17. The kit according to any one of items 14 to 16, wherein the active ingredient is a free acid, a free base, a salt (such as hydrochloride, sulfate, phosphate, fumarate, tartrate, hemifumarate, maleate, aspartate, palmitate, tartrate, bitartrate, methanesulfonate, citrate, toluenesulfonate, bromate, sulfite, carbonate, bicarbonate, malate, succinate, ethanesulfonate, acetate, formate, sodium salt, iron salt, potassium salt, calcium salt) salt, ammonium salt or magnesium salt), or a co-crystal (the co-crystal is a co-crystal prepared from the active ingredient and a co-crystal ligand, and the co-crystal ligand used is natural amino acids, caffeine, saccharin, fumaric acid, tartaric acid, oxalic acid, malonic acid, maleic acid, aspartic acid, tartaric acid, bitartrate, pyroglutamic acid, nicotinic acid, nicotinamide, an organic base such as meglumine, tromethamine, isopropanolamine), or a hydrate, solvate, or solvate hydrate of the free acid, free base, salt or co-crystal.

[0090] 18. The kit according to any one of items 14 to 17, wherein the package 4 further comprises:

[0091] (1) pH regulator,

[0092] (2) Surfactants and / or cosurfactants, such as gum arabic, gum tragacanth, gelatin, propylene glycol monostearate, glyceryl monostearate, ethylene glycol distearate, diglyceryl monooleate, sodium lauryl sulfate, sorbitan fatty acid (Span 20, Span 40, Span 60, Span 65, Span 80, Span 83 or Span 85), potassium oleate, sodium oleate, triethanolamine oleate, lecithin, sucrose esters, polyoxyethylene-polyoxypropylene copolymer (poloxamer), propylene laurate, polysorbate (Tween 20, Tween 40, Tween 60, Tween 65, Tween 80 or Tween 85), polyoxyethylene monostearate, polyoxyethylene stearate, polyoxyethylene 400 monolaurate, polyoxyethylene 400 monostearate, polyoxyethylene 400 monooleate, polyoxyethylene lauryl ether, polyoxyethylene cetyl alcohol ether, vitamin E polyethylene glycol succinate (TPGS), betaine, polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, glycerin, sorbitol, polyethylene glycol, one or more; and / or

[0093] (3) Bile salts, such as sodium glycocholate, sodium deoxyglycocholate, sodium chenodeoxyglycocholate, sodium ursodeoxyglycocholate, sodium taurocholate, sodium taurodeoxycholate, sodium taurochenodeoxycholate, and sodium deoxycholate.

[0094] 19. A supramolecular self-assembled complex comprising the carrier I, carrier II and polymer as defined in any one of items 1 to 13, and preferably further comprising the active ingredient as defined in any one of items 14 to 17.

[0095] 20. A pharmaceutical composition comprising the supramolecular self-assembled complex according to item 19, and a pharmaceutical excipient selected from the group consisting of a filler, a disintegrant, a binder, a lubricant, a glidant, an emulsifier, a flavoring agent or a taste-masking agent, a surfactant, a co-surfactant, a preservative, a propellant, a pH regulator, an osmotic pressure regulator, a gel / ointment base, a suspending agent, a coating component, a pressure-sensitive adhesive, a penetration enhancer, and a combination thereof.

[0096] 21. The pharmaceutical composition according to item 20, which is in the form of a tablet, capsule, suspension, solution, patch, cream, gel, emulsion, eye drops, injection, oral capsule, suppository, implant, powder, granule or implant product.

[0097] 22. A method for preparing the supramolecular self-assembly complex described in item 19, comprising dissolving and mixing the carrier I, carrier II and polymer defined in any one of items 1 to 14, and the active ingredient defined in any one of items 14 to 17, respectively, wherein the carrier I and carrier II are added and mixed simultaneously, or are mixed separately with the polymer and / or the active ingredient and then mixed together.

[0098] 23. A composition for delivering an active ingredient, comprising the supramolecular self-assembled complex defined in item 19, and a pharmaceutically or food-acceptable carrier, excipient, diluent, adjuvant, vehicle, or a combination thereof.

[0099] 24. Use of the kit according to any one of items 1 to 18 or the supramolecular self-assembled complex according to item 19 for delivering an active ingredient, preferably for delivering to a mammal (such as a human, rat, mouse, or primate).

[0100] The high molecular weight polymer, surfactant and pH regulator used in the present invention are as follows:

[0101] (1) High molecular weight polymers

[0102] The polymer used in the present invention is a polymer commonly used in the art, such as cellulose (such as hydroxypropyl methylcellulose (HPMC), low-substituted hydroxypropyl cellulose (L-HPC), hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate (HPMCAS), carboxymethyl ethyl cellulose, sodium carboxymethyl cellulose (CMC-Na), hydroxyethyl cellulose, cellulose acetate phthalate); synthetic polymers such as polyethylene glycol (PEG), polyvinyl alcohol (PVA), polyvinyl pyrrolidone (PVP), copovidone (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), siRNA, mRNA; natural polymers and polysaccharide derivatives such as carrageenan, guar gum, gum arabic, locust bean gum, konjac gum, agar, gelatin, pectin, gellan gum, hyaluronic acid (HA), aminoglucosan, chondroitin sulfate, dermatan sulfate, heparin, keratan sulfate, heparan sulfate, sodium polyacrylate, sodium alginate, propylene glycol alginate, agar, fucoidan, cyclodextrin and derivatives, chitosan and derivatives (such as acylation, carboxylation, alkylation and quaternization, etc.), soy protein, plant protein, bovine serum albumin.

[0103] (2) Surfactants

[0104] The surfactant used in the present invention is a surfactant commonly used in the art, such as gum arabic, tragacanth gum, gelatin, propylene glycol monostearate, glyceryl monostearate, ethylene glycol distearate, diglyceryl monooleate, sodium lauryl sulfate, fatty acid sorbitan (Span20, Span40, Span60, Span65, Span80, Span83 or Span85), potassium oleate, sodium oleate, triethanolamine oleate, lecithin, sucrose esters, polyoxyethylene-polyoxypropylene copolymer (polo Sham), Propylene Glycol Laurate, Polysorbate (Tween 20, Tween 40, Tween 60, Tween 65, Tween 80 or Tween 85), Polyoxyethylene Monostearate, Polyoxyethylene Stearate, Polyoxyethylene 400 Monolaurate, Polyoxyethylene 400 Monostearate, Polyoxyethylene 400 Monooleate, Polyoxyethylene Laureth, Polyoxyethylene Cetyl Alcohol, Vitamin E Polyethylene Glycol Succinate (TPGS), Betaine, Polyoxyethylene Castor Oil, Polyoxyethylene Hydrogenated Castor Oil, Glycerin, Sorbitol, Polyoxyethylene Glycol.

[0105] (3) pH regulator

[0106] The pH regulator used in the present invention is a pH regulator commonly used in the art, for example, the acidic regulator is selected from taurine, acidic amino acids, hydrochloric acid, acetic acid, citric acid, tartaric acid, malic acid, lactic acid, ascorbic acid, phosphoric acid, adipic acid, fumaric acid, sulfuric acid, nitric acid, p-toluenesulfonic acid or succinic acid and aqueous dilutions of the above acids; the alkaline regulator is selected from basic amino acids, sodium hydroxide, sodium citrate, potassium citrate, monosodium citrate, sodium lactate, calcium hydroxide, potassium hydroxide, sodium bicarbonate, sodium carbonate, potassium bicarbonate, potassium carbonate, tromethamine, meglumine, sodium phosphate, disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium phosphate, dipotassium hydrogen phosphate, potassium dihydrogen phosphate, sodium acetate or potassium acetate or aqueous solutions of the above alkaline regulators; the neutral regulator is sodium chloride or potassium chloride.

[0107] The supramolecular self-assembling complexes provided by the present invention can be prepared into pharmaceuticals, health products, or diagnostic imaging agents suitable for mammalian medical and dietary supplement applications. The supramolecular self-assembling complexes can be conventionally presented in unit dosage forms and prepared by any method used in pharmaceutical, health product, or medical products. The supramolecular self-assembling complexes of the present invention can be formulated with other pharmaceutically or food industry-acceptable excipients or additives into dosage forms suitable for oral delivery, oral gastrointestinal delivery, rectal delivery, topical delivery, transdermal delivery, nasal delivery, diagnostic imaging agent delivery, and implantable delivery.

[0108] Based on this, the present invention provides a delivery system comprising the supramolecular self-assembled complex and a pharmaceutically or food-acceptable carrier, excipient, diluent, adjuvant, vehicle or a combination thereof.

[0109] The dosage form of the delivery system can be selected from oral solid preparations, liquid preparations, suspensions, granules, powders, dry suspensions, etc.; oral patches, sublingual tablets, lozenges, rapidly disintegrating tablets, orally disintegrating films, etc.; external patches, gels, creams, emulsions, sprays, etc.; ointments, emulsions, gels, implants, etc.; suppositories, gels, ointments, etc. for intracavitary mucosal delivery; solutions, powder injections, oil injections, etc. for injection routes; and suitable dosage forms well known to those skilled in the art.

[0110] In some embodiments, the oral solid preparation is selected from ordinary tablets or capsules, sustained-release tablets or sustained-release capsules, controlled-release tablets or capsules, granules, suspensions, solutions, films, orally disintegrating tablets, orally disintegrating films, sublingual tablets, capsule patches, etc.

[0111] Specifically, for oral therapeutic or health care administration, it can be combined with one or more other excipients and used in the form of swallowable tablets, buccal tablets, sugar-coated tablets, capsules, elixirs, suspensions, syrups, powder tablets, etc. Excipients may be (but are not limited to): binders (such as hydroxypropyl cellulose, povidone or hydroxypropyl methylcellulose); fillers (such as microcrystalline cellulose, resistant starch, resistant dextrin, pregelatinized starch, starch, mannitol or lactose); disintegrants (such as croscarmellose sodium, crospovidone or sodium starch glycolate); lubricants (such as magnesium stearate, stearic acid or other metal stearates); sweeteners (such as sucrose, fructose, lactose or aspartame); and / or flavorings (such as peppermint, wintergreen oil or cherry flavoring), pH adjusters (such as hydrochloric acid, sulfuric acid, citric acid, sodium (potassium) hydroxide, sodium bicarbonate, disodium hydrogen phosphate, sodium dihydrogen phosphate, ammonia, triethanolamine, tromethamine, etc.), antibacterial agents (such as benzyl alcohol, phenoxyethanol, benzoic acid, sodium benzoate, sorbic acid / potassium sorbate, methylisothiazolinone, parabens, pentylene glycol, hexylene glycol, caprylyl glycol and ethylhexylglycerin, p-hydroxyacetophenone and some natural extracts).

[0112] When unit dosage form is a capsule, in addition to the material of the above type, it can also contain a liquid carrier, such as vegetable oil or polyethylene glycol. Various other materials can be in coated form or in other forms that change the physical form of the solid unit dosage form. For example, tablets, pills or capsules can be coated with gelatin, polymers, wax, shellac or sugar. Of course, any material for the preparation of any unit dosage form will typically be pharmaceutically acceptable and nontoxic in fact under the amount adopted.

[0113] Formulations for rectal administration may be presented as a suppository with a suitable base comprising for example cocoa butter or a salicylate.

[0114] The target delivery substance, carrier, and ratio in the supramolecular self-assembly complex of the present invention can be calculated and optimized based on the effective dosage of the delivery substance in different application scenarios, the application method, the safe dosage of the carrier, the physicochemical properties, and chemical structure of the target delivery substance and the carrier, and then the optimal formula composition can be determined in combination with in vitro or in vivo experiments, in vivo experiments, and human trials.

[0115] The term "about" used herein refers to a set of all values ​​within a range of ±5% of the numerical value given thereafter.

[0116] The unexpected effects achieved by the present invention are:

[0117] (1) Compared with the existing technology, the drug loading capacity is greatly improved;

[0118] (2) By introducing adaptive carriers, the penetration efficiency of target delivery substances throughout the gastrointestinal tract is greatly improved;

[0119] (3) Glycoside carrier conjugates significantly increase the water solubility of the carrier, allowing some drugs whose absorption is limited by dissolution rate to be developed into fast-acting emergency drugs;

[0120] (4) The interaction strength between the glycoside carrier and the target delivery substance is improved, so that the constructed supramolecular self-assembled complex has an electrostatic strong connection on the basis of the original loose connection, which improves the integrity and stability of the self-assembled complex in complex delivery scenarios and successfully solves the drug leakage caused by lipid nano-delivery and the sudden release of drugs caused by molecular accumulation during production or storage;

[0121] (5) Changing the effects of the physical and chemical properties of drug structure on drug absorption, transport, metabolism, and elimination, thereby better exerting the efficacy and toxic side effects of drugs;

[0122] (6) The supramolecular self-assembly complex provided by the present invention solves the clinical limitation that lipid nano-delivery and solid lipid nano-delivery cannot withstand the damage or metabolism of the complex gastrointestinal environment to the delivery system and must be administered by injection;

[0123] (7) It has higher drug loading capacity and better physical and chemical stability, thereby reducing the production cost of drugs;

[0124] (8) Through in situ assembly in vivo, the absorption window of the drug during gastrointestinal transit is expanded, the drug delivery efficiency and onset time are improved, and a more reliable long-term release and long-term absorption is provided;

[0125] (9) Improved patient compliance, medication safety and effectiveness;

[0126] (10) By improving drug delivery efficiency, the clinical dosage of drugs used by patients can be greatly reduced, thereby significantly reducing the toxic and side effects associated with drug dosage;

[0127] (11) The preparation process is simple, reliable, and easy to industrialize. Description of the drawings:

[0128] FIG1 is a HPLC chart of N-(R, S) chirality of naringenin in Example 1.

[0129] FIG2 is a NS chiral HPLC chart of naringenin in Example 1.

[0130] FIG3 is a chiral HPLC chart of naringenin NR in Example 1.

[0131] Figure 4 is the mass spectrum of the compound RBDS-B obtained in Example 2 under LC-MS / MS negative ion mode

[0132] Figure 5 shows the compound RBDS-B obtained in Example 2 1 H-NMR spectrum

[0133] Figure 6 is the mass spectrum of the compound STVB obtained in Example 2 under LC-MS / MS negative ion mode

[0134] Figure 7 shows the compound STVB obtained in Example 2 1 H-NMR spectrum

[0135] FIG8 is a mass spectrum of the compound RBDS-B-His obtained in Example 3 under LC-MS / MS negative ion mode;

[0136] Figure 9 shows the compound RBDS-B-His obtained in Example 3 1 H-NMR spectrum

[0137] Figure 10 is a mass spectrum of the compound STVB-Arga obtained in Example 4 under LC-MS / MS negative ion mode;

[0138] Figure 11 shows the compound STVB-Arga obtained in Example 4 1 H-NMR spectrum;

[0139] FIG12 is a mass spectrum of the compound STVB-His obtained in Example 4 under LC-LC-MS / MS negative ion mode;

[0140] Figure 13 shows the compound STVB-His obtained in Example 4 1 H-NMR spectrum;

[0141] FIG14 is a comparison of supramolecular self-assembled complexes Papp constructed with different formulations of paclitaxel and docetaxel in Example 8 (n=3)

[0142] Figure 15 is a particle size test report of the paclitaxel FF22 formulation diluted 200 times in Example 8

[0143] FIG16 is a particle size test report of the docetaxel FF32 formulation diluted 200 times in Example 8

[0144] FIG17 is a comparison of the dissolution of the supramolecular self-assembled complex of Apaxaban and the N-1 crystal form in Example 9

[0145] Figure 18 shows the detection amounts of nintedanib prototype drug and lipid metabolite BIBF1202 in liver tissue of rats 4 hours after oral gavage of the nintedanib supramolecular self-assembled complex and the reference preparation in Example 10. Specific implementation method:

[0146] To make the objectives, technical solutions, and advantages of the present invention more clear, the technical solutions of the present invention will be clearly and completely described in conjunction with the accompanying drawings. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts shall fall within the scope of protection of the present invention.

[0147] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in this field or in the product instructions were used. Instruments used, for which the manufacturer is not specified, are all conventional products that can be purchased through regular channels. The methods described are conventional methods unless otherwise specified. The polymers, excipients, pharmaceutical excipients, pH adjusters, reagents, surfactants, and cosurfactants can all be purchased through regular channels and, unless otherwise specified, can be obtained from public commercial sources.

[0148] Reference Listed Drug (RLD) is a different drug that has been marketed and generally refers to a drug that has undergone complete safety and efficacy studies. For example, the RLD for ibrutinib should be Imbruvica Ibrutinib capsules (licensee: Pharmacyclics LLC).

[0149] To further illustrate the present invention, the supplier information of the target delivery agent (also referred to as active ingredient in the present invention), carrier, enzyme, etc. used in the supramolecular self-assembly complex provided by the present invention is summarized as follows in conjunction with the examples:

[0150] Nintedanib, ibrutinib, lenvatinib, dabigatran etexilate, S-naringenin, R-naringenin, ezetimibe, RBDS-C-acid, OMGSD-V, MGSD-1E1, RBDS-B and its sodium salt, STVB and its sodium salt, DCS-A, and RBSD were prepared or isolated by Beijing Ruichuang Kangtai. Nilotinib, rivaroxaban, ticagrelor, olaparib, palbociclib, macitentan, acalabrutinib, posaconazole, MRTX1133, ketoconazole, itraconazole, and celecoxib were purchased from Pinghu Aibo Chemical Co., Ltd.; apixaban was purchased from Suzhou Capewell Pharmaceutical Technology Co., Ltd.; paclitaxel and docetaxel were purchased from Fujian Nanfang Pharmaceutical Co., Ltd.; cannabidiol was purchased from Yunnan Gongma Biotechnology Co., Ltd.; and naringenin racemate and bis(4-nitrobenzene) phosphate were purchased from Beijing Bailingwei Technology Co., Ltd.

[0151] The CypExpress™ 3A4 enzyme (humanized) kit was purchased from SIGMA ALDRICH.

[0152] The raw materials, auxiliary materials, reagents and solvents such as the carrier, high molecular polymer and pH adjuster used in the present invention are not particularly limited, and commercially available conventional raw materials, auxiliary materials, reagents and solvents can be used.

[0153] Some of the vectors used in the present invention are currently not commercially available, or the preparation methods of the vector derivatives designed and developed by the inventors will be clearly described in the present invention.

[0154] The preparation methods of the media used in the present invention, such as the dissolution medium and the disintegration medium, are prepared in accordance with the "Technical Guidelines for Dissolution Testing of Ordinary Oral Solid Preparations", while taking into account the preparation methods specified in the four general rules of the 2020 edition of the Chinese Pharmacopoeia and the current edition of the United States Pharmacopoeia, and are not specifically listed here.

[0155] The preparation methods of the pre-meal simulated gastric fluid (Fasted State Simulated Gastric Fluid, FaSSGF), pre-meal simulated intestinal fluid (Fasted State Simulated Intestinal Fluid, FaSSIF), post-meal simulated gastric fluid (Fed State Simulated Gastric Fluid, FeSSGF), and post-meal simulated intestinal fluid (Fed State Simulated Intestinal Fluid, FeSSIF) used in the present invention are as follows:

[0156] (1) Preparation of simulated gastric juice to simulated intestinal juice before meals

[0157] Preparation of FaSSGF: Weigh 2g NaCl, 0.043g sodium taurocholate, and 0.015g lecithin into approximately 800ml of degassed purified water and dissolve them by ultrasonic stirring (heating at 40-50°C). After the lecithin is completely dissolved, adjust the pH to 1.60 with 1mol / L HCl. Then, weigh 0.1g pepsin, dilute to 1000ml, and adjust the pH to 1.6.

[0158] FaSSIF-V2 preparation: 187 ml FaSSGF + 63 ml conditioning solution (8.8056 g maleic acid, 8.8062 g NaOH, 9.9762 g NaCl, 6.618 g sodium taurocholate, and 0.5508 g lecithin are weighed and dissolved in 1000 ml degassed purified water, stirring ultrasonically (heating at 40-50°C)). If necessary, adjust the pH to pH 6.5 with hydrochloric acid or sodium hydroxide solution.

[0159] (2) Preparation of simulated gastric fluid after meal to simulated intestinal fluid

[0160] Preparation of FeSSGF: Weigh 13.85g NaCl, 2.44g anhydrous sodium acetate, and 1.028g glacial acetic acid and dissolve them in 1000ml degassed purified water. Then add 1000ml milk and mix well. Adjust the pH to 5.00 with concentrated hydrochloric acid.

[0161] Preparation of FeSSIF-V2: To 150 ml of FessGF+, add 100 ml of conditioning solution (weigh 15.9654 g maleic acid, 8.165 g NaOH, 7.9481 g NaCl, 13.75 g sodium taurocholate, 0.6089 g sodium oleate, and 3.79 g lecithin in 1000 ml of degassed purified water and dissolve under ultrasonic stirring). If necessary, adjust the pH to 5.8 with hydrochloric acid or sodium hydroxide solution.

[0162] The assay method used in the present invention is:

[0163] (1) Nanoparticle size, PDI:

[0164] The particle size was determined using a NanoBrook (model: 90Plus PALS) nanoparticle size analyzer. Dynamic light scattering was used at room temperature (25°C) and a diffraction angle of 90°. Prior to measurement, the supramolecular self-assembled complex solution was diluted to 100 μg / ml. 50 μl was dispersed in 1 ml of deionized water and homogenized by sonication. The measurement was immediately performed for 2 minutes, with three replicates per sample.

[0165] (2) Zeta potential determination:

[0166] Measurements were performed at room temperature (25°C) with the zeta potential diffraction angle set at 15°C. Prior to measurement, the supramolecular self-assembled complex solution was diluted to 100 μg / mL. 50 μL of the solution was dispersed in 1 mL of phosphate buffer at different pH values ​​(6.0, 7.0, 7.4, and 8.0) for measurement. Three replicates were performed for each sample. 1 mM phosphate buffer was prepared with ultrapure water and the pH was adjusted with 0.1 N hydrochloric acid or sodium hydroxide solution. The sample should be transparent; concentrated samples can be centrifuged and the supernatant collected.

[0167] (3) Determination of active ingredient content:

[0168] The determination was performed by high performance liquid chromatography (HPLC) with an external standard method. The chromatographic conditions are shown in Table 1.

[0169] (4) Determination of active ingredient degradation products:

[0170] For different drugs, the high performance liquid chromatography method developed by the inventors was used for determination, as shown in Table 1.

[0171] (5) Calculation of Encapsulation Efficiency (EE):

[0172] When preparing supramolecular self-assembled complexes, stir at room temperature to remove the organic solvent, centrifuge at 15,000 rpm for 10-20 minutes to remove the precipitate and larger particles, take a certain volume of supernatant, add an excess of organic solvent, such as chromatographic grade methanol, ethanol, acetonitrile, and sonicate to completely release the drug from the carrier, and determine the content using a high performance liquid chromatography according to the chromatographic conditions under the target delivery object, and then calculate according to the following formula.

[0173] EE = [encapsulated drug amount (mg) / initial added amount (mg)] × 100%.

[0174] (6) Drug loading determination:

[0175] A certain volume of the clarified supramolecular self-assembled complex solution after centrifugation was slowly evaporated to dryness at 40°C, and the solid content per unit volume (g / ml) after concentration was weighed. Drug loading % = drug content per unit volume (mg) / solid content per unit volume (mg / ml) * 100.

[0176] Naringenin is a reported inhibitor of cytochrome 3A4 (CYP3A4). Commercially available naringenin (N) is a racemic form, while single-configuration R-naringenin (RN) and S-naringenin (SN) are prepared using chiral preparative columns using high-pressure preparative liquid chromatography or supercritical fluid chromatography.

[0177] Table 1: Determination methods of delivery target content and related substances

[0178] Instruments and methods used for collecting data on the structural characterization of each derivative of the present invention:

[0179] (1) Proton nuclear magnetic resonance (1HNMR) data were collected using a Bruker AVANCE 600 instrument with a resonance frequency of 600 MHz and deuterated DMSO as the solvent.

[0180] (2) The high-resolution mass spectrometry data of the present invention were detected using a Waters Xevo TQ-S triple quadrupole mass spectrometer, and the liquid chromatography conditions were:

[0181] The chromatographic column was Agilent ZOBAX SB-C18, 2.1 × 50 mm, 1.8 μm;

[0182] Mobile phase A: 0.1% formic acid-water solution; Mobile phase B: 0.08% formic acid-acetonitrile solution;

[0183] Detection wavelength: 210nm;

[0184] Flow rate: 0.7 mL / min;

[0185] Injection volume: 5 μL;

[0186] Column temperature: 60°C;

[0187] 1) The gradient elution procedures for RSDB-B-Na, STVB-Arga, and STVB-His are as follows:

[0188] 2) The gradient elution procedure for RBDS-B-His is:

[0189] 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.

[0190] (3) The purity of each derivative was tested by high performance liquid chromatography, and the chromatographic conditions were as follows:

[0191] (4) Determination of sodium content in RBDS-B-Na and STVB-Na

[0192] The sodium salt content was titrated using a potentiometric titrator (Metrohm 916-Ti Touch automatic potentiometric titrator) as follows:

[0193] Sodium content determination: Dissolve approximately 0.4 g of this product in 50 ml of ethanol-water (50:50) and titrate with 0.1 mol / L hydrochloric acid solution according to the potentiometric titration method (Chinese Pharmacopoeia 2020 Edition, Part IV, General Rules 0701). Correct the titration result with a blank test. Each 1 ml of 0.1 mol / L hydrochloric acid solution is equivalent to 2.30 mg of sodium. Calculate the sodium content based on the anhydrous form.

[0194] 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.

[0195] 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.

[0196] Example 1: Chiral Preparation of R- and S-Ningenin

[0197] Table 1 shows the chiral analysis method for naringenin (NRG). Chiral HPLC analysis of the purchased racemate revealed that the S- and R-configurations accounted for 47.66% and 52.35%, respectively. The peak in the chiral analysis HPLC chromatogram with a retention time of approximately 4.7 min was the S-configuration, and the peak with a retention time of 6.5 min was the R-configuration. Chiral separation was performed using a Thar 80 preparative supercritical fluid chromatograph with a Chiralpak AD-H 250 mm × 30 mm, 5 μm column and a detection wavelength of 214 nm. Mobile phases A and B consisted of supercritical carbon dioxide and isopropanol, respectively. Elution was performed using 40% isopropanol isocratic elution at a flow rate of 70 g / min. Continuous injections were made with a 5 ml injection volume and a sample concentration of 800 mg / ml. Fractions from the two peaks were collected, combined, and dried on a rotary evaporator in a 40°C water bath. After spin drying, the R- and S-configuration naringenin were re-analyzed by chiral HPLC, and the ee values ​​were both greater than 98%. The chiral HPLC spectra before separation and the chiral purity confirmation HPLC spectra of the R- and S-configuration after separation are shown in Figures 1 to 3, respectively.

[0198] CYP3A4 enzyme activity confirmation

[0199] Enzyme activity was confirmed according to the instructions provided on the Sigma official website. The results are shown in Table 2 below:

[0200] Table 2. Enzyme activity results

[0201] Based on the above results, it was confirmed that the enzyme activity met the requirements for use.

[0202] Example 2. Preparation of RBDS-B, STVB sodium salt or potassium salt:

[0203] (1) Preparation of RBDS-B or RBDS-B-Na:

[0204] In a 250 mL three-necked flask, 25 g of rebaudioside A (RBDS-A) and 180 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 100 mL of purified water. The filter cake was collected and dried under reduced pressure at 40° C. to obtain 20 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 4. 1H 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),4.442-4.496(m,2H). .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 5 for details.

[0205] In a 500-mL three-necked flask, 28.00 g of RBDS-B was dispersed in 280 mL of purified water. 1.39 g of solid sodium hydroxide was added in batches at a temperature below 40°C, stirred to dissolve, and filtered. 560 ml of anhydrous ethanol was added to the filtrate, which was then cooled 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 22 g of RBDS-B-Na as an off-white solid with an HPLC purity of 96.7% and a sodium content of 2.77%.

[0206] (2) Preparation of STVB or STVB-Na:

[0207] In a 1L three-necked flask, 50g of stevioside (STVS) and 400ml 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 200mL of purified water, and the filter cake was collected and dried under reduced pressure at 40°C to obtain 36g 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 positive and negative ion modes were 660.3461 (M+H2O)+ / 641.0961 (MH), respectively. - , the high-resolution mass spectrometry negative ion mode spectrum is shown in Figure 6, 1H 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),3.18 4-3.117(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 7 for details.

[0208] In a 1L three-necked flask, 18.00g of STVB was dispersed in 180mL of purified water. 0.9g of solid sodium hydroxide was added in batches at a temperature below 40°C, stirred until dissolved, and filtered. 360ml of anhydrous ethanol was added to the filtrate, which was then cooled 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 20g of STVB-Na as an off-white solid with an HPLC purity of 97.4% and a sodium content of 3.46%.

[0209] The contents of RBDS-B(Na) and STVB-(Na) were determined by high performance liquid chromatography. The chromatographic conditions are shown in Table 3 below:

[0210] Table 3. Chromatographic conditions for the assay method are as follows:

[0211] Using the above method, the liquid chromatography purities of RBDS-B and STVB were 96.7% and 97.4%, respectively.

[0212] The sodium salt contents of RBDS-B and STVB were 2.77% and 3.46% respectively, as determined by potentiometric titration.

[0213] Determination of solubility of RBDS-B, STVB free acid and sodium salt:

[0214] Take appropriate amounts of RBDS-B, STVB free acid and RBDS-B, STVB sodium salt, add pH 1.0 hydrochloric acid, pH 4.0 acetate buffer, pH 6.8 phosphate buffer, water, and pH 7.4 phosphate buffer, respectively, shake in a 25°C constant temperature air bath shaker, centrifuge at 25°C and 13000 rpm / min for 10 minutes at 12 h and 24 h, respectively, take the supernatant for analysis, and perform detection using the method in Table 3. The results are shown in Table 4 below:

[0215] Table 4. Solubility study (25℃) Note: “—” solubility was too high and was not further developed.

[0216] The results of solubility experiments show that after RBDS-B and STVB are made into sodium salts, their solubility is higher under physiological pH conditions and they are more suitable for use as delivery carriers.

[0217] Example 3. Preparation and characterization of RBDS-B-His

[0218] The preparation of RBDS-B was the same as in Example 2.

[0219] (1) Synthesis of RBDS-B-Ac-Cl

[0220] In a 500 mL three-necked flask, 18 g of rebaudioside B (RBDS-B), 45 g of acetic anhydride and 4.5 g 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, 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 25 g of RBDS-B-Ac as a light yellow solid with a purity of 97.5% as determined by HPLC.

[0221] In a 500 mL three-necked flask, add 20 g of RBDS-B-Ac and 300 mL of dichloromethane, stir the reaction system until dissolved, add 1 drop of N,N-dimethylformamide, and then add 3.1 g 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 21 g of RBDS-B-Ac-Cl as a light yellow solid.

[0222] (2) Preparation and characterization of RBDS-B-His

[0223] 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. The reaction system was stirred at room temperature for 4 hours. The reaction system was washed three times with 350mL of purified water. 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.

[0224] 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 and reacted at room temperature for 4 hours. The mixture 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 negative ion mode was 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 8. 1H NMR (600MHz, d-DMSO) δ8.239(s,1H),7.718-7.570(m,1H),7.233-7.186(m,1H),7.014-6.982(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 9 for details; HPLC purity is 99.09%.

[0225] Example 4. Preparation and Characterization of STVB-Arga and STVB-His

[0226] (1) The preparation of STVB is the same as that in Example 2.

[0227] (2) Preparation of STVB-Ac-Cl

[0228] In a 500mL three-necked flask, 18g of STVB, 42g of acetic anhydride and 5.5g of sodium acetate were added, the reaction system was heated to 140°C, stirred and reacted for 6 hours, the reaction system was cooled to room temperature, 400mL of dichloromethane was added, the organic phase was washed three times with 200mL of purified water, the organic phase was collected and dried under reduced pressure at 40°C to obtain 23g of STVB-Ac as a light brown solid.

[0229] In a 500mL three-necked flask, add 23g of STVB-Ac and 300mL of dichloromethane, stir the reaction system until dissolved, add 1 drop of N,N-dimethylformamide, and then add 4.6g 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 21g of STVB-Ac-Cl as a light brown foamy solid.

[0230] (3) Preparation and characterization of STVB-Arga

[0231] 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.

[0232] 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 main peak negative ion mass-to-charge ratio was 796.1407 (MH)- and the adduct peak mass-to-charge ratio was 841.9503 (M+2Na-H)-, which was consistent with the theoretical accurate molecular weight of the main peak. The high-resolution mass spectrometry negative ion mode spectrum is shown in Figures 10 and 1H. 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 11 for details, HPLC purity is 97.82%.

[0233] (4) Preparation and characterization of STVB-His

[0234] 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.

[0235] 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 main peak negative ion mass-to-charge ratio was 777.0210 (MH) - and combined peak 823.0913 (M+2Na-H) - , the high-resolution mass spectrometry negative ion mode spectrum is shown in Figure 12; 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 13 for details, HPLC purity is 97.64%.

[0236] Example 5. Evaluation of STVB-His, STVB-Arga, and RBDS-B-His Characteristics

[0237] (1) Solubility determination

[0238] Take appropriate amounts of STVB-His, STVB-Arga and RBDS-B-His derivatives and conduct solubility test according to the solubility test method of Part IV, General Rules 15 (2) of the 2020 edition of the Chinese Pharmacopoeia.

[0239] Weigh each derivative ground into fine powder, place it in a certain amount of solvent at 25℃±2℃, shake it vigorously for 30 seconds every 5 minutes, and observe the dissolution within 30 minutes. If no solute particles are visible, it is considered to be completely dissolved.

[0240] The assay results showed that RBDS-B-His was soluble in water and methanol, slightly soluble in 95% ethanol and pH 6.8 phosphate buffer, and almost insoluble in acetone;

[0241] STVB-Arga is freely soluble in water and pH 6.8 phosphate buffer, slightly soluble in methanol, slightly soluble in 95% ethanol, and almost insoluble in acetone;

[0242] STVB-His is slightly soluble in water, slightly soluble in methanol, 95% ethanol and pH 6.8 phosphate buffer, and almost insoluble in acetone.

[0243] (2) Stability study in pre-meal simulated gastric fluid (FaSSGF) and pre-meal simulated intestinal fluid (FaSSIF-V2)

[0244] Take an appropriate amount of RBDS-B-His and STVB-Arga and add 12 ml of the above-mentioned gastric juice or 20 ml of intestinal fluid, respectively, to prepare a solution containing approximately 0.5 mg of each vector per 1 ml. Ultrasonicate at 37°C to evenly disperse the solution. Place the solution in a 37°C constant-temperature air shaker at 250 rpm, with the shaker set to time 0. Samples were collected and tested for concentration at different time points according to the table below.

[0245] Pre-prandial gastric and intestinal fluid samples: Take 3 ml from each sampling point, centrifuge at 37°C and 15,000 rpm for 5 minutes, take 0.5 ml of the supernatant, add 1.5 ml of acetonitrile, mix well, filter through a 0.22 μm nylon filter, discard 1 ml of the initial filtrate, and analyze the subsequent filtrate by HPLC. Place in a 10 ml volumetric flask, add 2.0 ml of artificial gastric or artificial intestinal fluid, quickly sonicate to disperse evenly, immediately place in a 37°C constant temperature oscillator, shake at 200 rpm. At the sampling time, add an appropriate amount of acetonitrile, sonicate for 10 minutes, dilute to the mark with acetonitrile, centrifuge at 15,000 rpm for 5 minutes, take 0.5 ml of the supernatant and add 0.5 ml of water, mix well, centrifuge at 15,000 rpm for 10 minutes, and use the supernatant as the test solution.

[0246] Sampling points in simulated gastric fluid before meal: 0.5h, 1h; Sampling points in simulated gastric fluid after meal: 1h, 2h;

[0247] Sampling points in simulated intestinal fluid before and after meal: 1h, 2h and 4h.

[0248] The content of the carrier conjugate was detected by HPLC external standard method. The results are shown in Table 5-6 below:

[0249] Table 5. Stability of each derivative in biological media

[0250] All carriers showed 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 little effect on the stability of the carriers.

[0251] Table 6. Stability of each derivative in biological media

[0252] Example 6. Preparation of supramolecular self-assembly complex:

[0253] In this embodiment, the expression "vector" refers to vector I and / or vector II, and the expression "partial vector" refers to vector I or vector II.

[0254] Method 1 (I):

[0255] ① Dissolve the active ingredient in a water-miscible organic solvent (such as a single solvent: methanol, ethanol, isopropanol, acetone, tetrahydrofuran, DMSO, etc.; or a mixed solvent: methanol-water, ethanol-water, ethanol-acetone, acetonitrile-water, methanol-tetrahydrofuran-water, etc.) and set aside;

[0256] ② Dissolve the carrier and high molecular polymer in an aqueous medium, such as water or buffer.

[0257] While stirring, slowly add ① to ② dropwise until visibly turbidity appears. Stop adding dropwise and continue stirring for 30 minutes at a temperature not exceeding 45°C. Recover the organic solvent using a rotary evaporator. Alternatively, continue stirring until the organic solvent odor disappears. Centrifuge or filter the remaining aqueous solution to obtain the supramolecular self-assembly solution. If desired, the solution can be freeze-dried, spray-dried, concentrated using a rotary evaporator, or dried by adding formulation excipients.

[0258] The carrier or polymer can also be dissolved in an organic solvent together with the active ingredient (when the carrier, part of the carrier, or the polymer is insoluble in the organic solvent, the carrier or part of the carrier or the polymer can be first dissolved in water or an aqueous medium and then mixed with the active ingredient solution).

[0259] Method 2(II):

[0260] The active ingredient and the carrier are dissolved in a solvent (depending on the solubility of the active ingredient and the carrier, a low-boiling-point organic solvent immiscible with water, such as dichloromethane, chloroform, ether, ethyl acetate, etc., a low-boiling-point organic solvent miscible with water, such as methanol, ethanol, acetone, isopropanol, etc., or a mixed solvent of two or three of the above, such as methanol-dichloromethane, ethanol-dichloromethane-water, etc.), and concentrated on a rotary evaporator to form a thin film. Then, an aqueous solvent, such as water or a buffer solution, is added and shaken for hydration. The resulting solution is centrifuged or filtered to obtain a supramolecular self-assembled complex solution.

[0261] Alternatively, part of the carrier that is insoluble in the organic solvent may be dissolved in the aqueous medium.

[0262] Method 3(III):

[0263] (1) The active ingredient and carrier are dissolved in an organic solvent containing an appropriate amount of water (such as methanol-water, ethanol-acetone-water, acetone-dichloromethane-water, etc.), the organic solvent is recovered by rotary evaporation, and the remaining aqueous solution is centrifuged or filtered to obtain a supramolecular self-assembled complex solution.

[0264] (2) The active ingredient and the carrier are dissolved in a solvent (depending on the solubility of the active ingredient and the carrier, a low-boiling point organic solvent that is immiscible with water, such as dichloromethane, chloroform, ether, ethyl acetate, etc., a low-boiling point organic solvent that is miscible with water, such as methanol, ethanol, acetone, isopropanol, etc., or a mixed solvent, such as methanol-dichloromethane, ethanol-dichloromethane-water, etc., can be selected).

[0265] After mixing (1) and (2), spray-dry, freeze-dry or concentrate to dryness using a rotary evaporator, or concentrate the organic solvent first and then freeze-dry.

[0266] Method 4 (IV):

[0267] The active ingredient and part of the carrier are dissolved in a water-immiscible organic solvent (such as dichloromethane, chloroform, ether, etc.) and set aside; the carrier or part of the carrier is dissolved in an aqueous medium, such as water or buffer; the two solutions are mixed, homogenized or high-pressure homogenized, and the organic solvent is recovered to obtain a supramolecular self-assembled complex solution.

[0268] Example 7. Preparation of supramolecular self-assembled complexes containing BTK inhibitors

[0269] Bruton's tyrosine kinase (BTK) inhibitors have shown significant clinical advantages in the treatment of B cell malignancies and some B cell immune diseases.

[0270] The supramolecular self-assembly complex was prepared using ibrutinib (IRB) and acalabrutinib (ACB), two BTK inhibitors, as active ingredients. The solution provided by the present invention is also applicable to other BTK inhibitors. The specific formulation process is as follows:

[0271] Table 7. BTK inhibitor supramolecular self-assembled complex aqueous solution and phenomena observed over 72 hours

[0272] (1) Preparation process

[0273] The active ingredient of formulas F1-F7 is ibrutinib, and the active ingredient of F8-F14 is acalabrutinib.

[0274] The active ingredient was dissolved in a methanol-tetrahydrofuran (1:2, V / V) mixed solvent to prepare a solution with a concentration of 0.4 g / ml;

[0275] Add 10 ml of water to the carrier and high molecular polymer in the amounts shown in Table 7 and dissolve them by ultrasonication. Among them, F14 could not be completely dissolved by adding 10 ml of water and ultrasonication for 20 minutes. It still did not dissolve after adding 5 ml of water. This formula was discarded. The other formulas could be dissolved and clarified.

[0276] Prepare supramolecular self-assembled complex solution according to method I. Stop adding dropwise when obvious turbidity appears and continue stirring for 30 min. Remove organic solvent using a rotary evaporator at 40°C. Centrifuge at 15,000 rpm for 5 min, separate the supernatant, and let it stand at room temperature for 72 h to observe whether there is precipitation, stratification, or turbidity.

[0277] (2) Dilution tolerance test

[0278] After 72 hours of stabilization at room temperature, 0.5 ml of a sample remained clear and was diluted with pH 6.8 phosphate buffer. The dilution was observed at different dilution ratios, and the sample was allowed to stand for 15 minutes after each dilution. The dilution results are shown in Table 8 below. The sample that remained clear after 100-fold dilution was assayed. For comparison, samples F1, F2, F3, and F4 were also stabilized for 72 hours and then re-centrifuged. The supernatant was then assayed.

[0279] Comparative Example 1: Ibrutinib and a high molecular weight polymer PVP K30 were dissolved in methanol at a mass ratio of 3:1 between the polymer and ibrutinib. The spray dryer was set to an inlet temperature of 120°C and an outlet temperature of 60°C with nitrogen as the carrier gas to prepare a solid dispersion. The obtained solid dispersion was dried in a vacuum drying oven for 4 hours. The content was detected by HPLC, and the drug loading was 25%.

[0280] Weigh an appropriate amount of the solid dispersion of Comparative Example 1, containing about 20 mg of ibrutinib.

[0281] Table 8. Dilution tolerance test of supramolecular self-assembled complexes of BTK inhibitors with different formulations Note 1: “--” The experiment was not carried out.

[0282] Ibrutinib and acalabrutinib have intrinsic solubilities below 0.01 mg / ml in aqueous solutions at pH 5.0-7.4, making them poorly soluble drugs. Using RBDS-A or RBDS-A in combination with the polymer PVP K30 increased their solubility by approximately 78-fold and 188-fold, respectively, in F1 and F3. The mass ratio of the active ingredient to the RBDS-A carrier was approximately 1:200 in F1, and approximately 1:18 in F3. Combinations of different chemical carriers increased the solubility of F7 and F8 by approximately 6318-fold (without the polymer) and 6919-fold (with the polymer), respectively. At these ratios, the active ingredient to carrier or polymer ratios were approximately 1:1.5-1.8 and 1:1.2-1.3, respectively. Combinations of acalabrutinib with different chemical carriers, F9 and F11, increased their solubility by 5761-fold (without the polymer) and 6626-fold (with the polymer), respectively. The combined use of carriers with different chemical structures, with or without the addition of high molecular weight polymers, is far more effective in improving the solubility of poorly soluble drugs than the use of a single carrier or a single carrier combined with a high molecular weight polymer.

[0283] (3) Characteristic evaluation

[0284] The particle size and Zeta potential of the supramolecular self-assembled complex prepared by F7-F11 were tested, and the encapsulation efficiency and drug loading were calculated. The remaining samples were freeze-dried in a freeze dryer, and the re-dissolution after freeze-drying (100-fold dilution of pH 6.8 phosphate) and the change in particle size after re-dissolution were investigated. The results are shown in Table 9 below.

[0285] Table 9. Performance evaluation of supramolecular self-assembled complexes of BTK inhibitors with different formulations

[0286] in conclusion:

[0287] Drug loadings for formulations F1-F4 were 0.48%, 0.95%, 1.13%, and 2.53%, respectively. Formulations F7 and F9, which combined carriers with different chemical structures, achieved drug loadings of 21.4% and 35.6%, respectively. Formulations F8 and F11, which combined carriers with different chemical structures and polymers, achieved drug loadings of 23.8% and 37.9%, respectively. Combining carriers with different chemical structures increased drug loading by 23% to 35% compared to either carrier alone. Combining carriers with polymers increased drug loading by 20% to 35% compared to single carrier and polymer combinations in F3 and F4. The combined use of carriers with different chemical structures was significantly superior to that of either a single carrier or a single carrier combined with a hydrophilic polymer. The average particle size of formulations F8 and F11 containing polymers was larger than that of formulations F7 and F9 containing only carriers, but the average particle size was still less than 100 nm.

[0288] (4) Chemical stability and physical stability

[0289] The above-mentioned ibrutinib supramolecular self-assembled complex was sealed with an aluminum-plastic composite film and placed under high temperature conditions of 50°C for influencing factor experiments. Samples were taken on the 14th and 30th days to detect related substances, and samples were taken on the 30th day for re-dissolution with pH 6.8 phosphate buffer, and the particle size was detected. At the same time, the chemical stability was compared with that of Comparative Example 1. The ibrutinib-related substance detection method in Table 1 was used for detection, and the results are shown in Table 10 below.

[0290] Table 10. Experimental results of factors affecting the supramolecular self-assembly complex of Ibrutinib Note 2: “ / ” indicates not detected or below the limit of quantification.

[0291] Conclusion: The results of related substance determination after 30 days of storage at 50°C in packaging indicate that the levels of degradation impurities ZZ-03, ZZ-05, ZZ-09, ZZ-03, ZZ-22, and ZZ-11 in ibrutinib F7 and F8 all increased slightly compared to day 0. The increase in each impurity in F7 formulation was slightly greater than that in F8 formulation, but still did not exceed the acceptable limit for degradation impurities. Ibrutinib prepared into supramolecular self-assembled complexes according to formulations F7 and F8 showed good chemical stability. Compared to the solid dispersion prepared from the polymer PVP K30 and ibrutinib in Comparative Example 1, after 30 days of storage at 50°C, the increases in ZZ-22, ZZ-11, the unknown single impurity RRT1.75, ZZ-09 and ZZ-03, and the total impurities were all significantly higher than those in the supramolecular self-assembled complex. According to the re-dissolution results and particle size measurement results of the supramolecular self-assembled complex samples placed at high temperature of 50°C for 30 days, the physical stability of the two formulations is also very good.

[0292] (5) Enzyme metabolic stability

[0293] The experiment was conducted according to the procedures specified in the CYP3A4 enzyme instructions. Itraconazole (ITCZ) was used as a positive inhibitor. Ibrutinib, acalabrutinib, and the positive inhibitor were poorly soluble in water and were initially dissolved in a small amount of dimethyl sulfoxide (DMSO) (no more than 2% in the final enzyme reaction system). The enzyme inhibitor was selected from racemic naringenin N(R,S), R-configuration naringenin (RN), and S-configuration naringenin (SN). The positive inhibitor was itraconazole (ITCZ). Preliminary experimental results showed that at 5 μM ibrutinib, a CYP3A4 enzyme concentration of 50 mg / mL, and an itraconazole concentration of 0.625 μM, approximately 50% enzyme activity was inhibited.

[0294] The substrate was ibrutinib, and the substrate concentration, reaction system enzyme concentration, and positive inhibitor concentration were based on the preliminary experimental results. A negative control group and a positive control group were set up to investigate the inhibitory effects of potential inhibitors N(R, S), RN, and SN on CYP3A4 enzyme. Each experiment was repeated three times.

[0295] The experimental design is shown in Table 11 below:

[0296] Table 11. Experimental design

[0297] The above experiment was carried out according to the following operating steps, as shown in Table 12 below.

[0298] Table 12. Experimental steps

[0299] According to the results of the localization experiment, itraconazole (ITCZ), blank solvent, potential enzyme inhibitors, and enzymes did not interfere with the detection of the ibrutinib peak, and the ibrutinib degradation product peak did not interfere with the detection of ibrutinib.

[0300] Acalabrutinib samples were prepared in the same manner.

[0301] Ibrutinib and acalabrutinib were assayed using the HPLC method shown in Table 1, and the results were calculated using the external standard method. Test solutions of Formulations F7 and F9 were prepared according to the measured contents. Three replicates were prepared for each formulation. The results are shown in Table 13 below.

[0302] Table 13. Experimental results (n=3)

[0303] In summary: From the negative control results, it can be seen that ibrutinib and acalabrutinib have good thermodynamic stability after incubation at 37°C for 4 hours.

[0304] The positive control was incubated at 37°C for 1 hour. The elimination rate constants of ibrutinib and acalabrutinib were 0.93 and 0.97 (h - 1 ), with elimination half-lives of 0.75 and 0.71 hours, respectively. After adding the positive inhibitor itraconazole and incubating at 37°C for 1 hour, the elimination half-lives of ibrutinib and acalabrutinib were 4.31 and 2.60 hours, respectively. The positive inhibitor significantly inhibited the activity of CYP3A4. The supramolecular self-assembled complex solution of ibrutinib and acalabrutinib was incubated with the CYP3A4 enzyme at 37°C for 1 hour. The elimination half-lives of ibrutinib and acalabrutinib were 5.27 (equivalent to the effect of 500μM inhibitor N) and 5.03 hours, respectively. The metabolic rate of the enzyme was significantly slowed, and the elimination half-life was significantly prolonged compared with the positive control and the positive inhibitor, and the metabolic stability of the enzyme was significantly improved.

[0305] (6) Pharmacokinetic study of oral ibrutinib supramolecular complex in beagle dogs

[0306] Pharmacokinetic study of different formulations of ibrutinib in beagle dogs after oral administration.

[0307] 1) Experimental drug preparation

[0308] Experimental drug: RLD (Ibrutinib Capsules, 140 mg) + NR: 250 mg of NR was added to each reference preparation, mixed evenly, and then filled into the capsules; other experimental drugs were directly converted from the ibrutinib supramolecular self-assembled complex formulas F7 and F8 into ibrutinib content and then filled into capsules.

[0309] 2) Dissolution assay

[0310] Method: Cuvette method, paddle method, speed: 100 rpm, temperature: 37°C, medium and volume: 187 mL FaSSGF / 250 mL FaSSIF-V2

[0311] The sampling points were: 15, 30, 45, 60, 90, 120, and 180 (min), and after 30 min of sampling, the fluid was adjusted to simulated intestinal fluid.

[0312] Sampling and sample preparation: 4 mL of sample was added to the dissolution vessel at 4°C and 4 mL of the same isothermal medium was added. The sample was centrifuged at 37°C, 15,000 rpm, and 5 minutes. 0.08 mL of the supernatant from the simulated gastric fluid was diluted with 3.52 mL of diluent. 0.1 mL of the supernatant from the intestinal fluid was diluted with 3.5 mL of diluent (50:50, V / V, MPA:acetonitrile). Ibrutinib content was determined using the method in Table 1. Dissolution results are shown in Table 14.

[0313] Table 14. Cumulative dissolution of supramolecular self-assembled complex tablets and reference formulations in FaSSGF→FaSSIF V2

[0314] Dissolution testing revealed that after switching from pre-meal simulated gastric fluid to pre-meal simulated intestinal fluid, the cumulative dissolution rates of F7 and FT formulations over 180 minutes were 90.5% and 99.2%, respectively, significantly higher than those of the reference formulation. The reference formulation had a 30-minute cumulative dissolution rate of 85.9% in pre-meal gastric fluid, significantly higher than those of F7 and F8. The cumulative dissolution rate of the reference formulation decreased significantly after switching to simulated intestinal fluid, but further dissolution occurred later, reaching a 180-minute cumulative dissolution rate of 75.7%. There was a slight difference in the cumulative dissolution rates of the F7 and F8 formulations in pre-meal gastric fluid. After switching to simulated intestinal fluid, the cumulative dissolution rate of F8 formulation over 180 minutes was approximately 9% faster than that of F7 formulation.

[0315] 3) Animal experiments

[0316] Dosage: 140 mg / animal for the reference preparation group and 140 mg / animal for the supramolecular self-assembled complex group, with crossover dosing. Washout period: 3 days.

[0317] Experimental animals: Adult male beagle dogs weighing 8-10 kg. All animals were fasted overnight (for more than 10 hours) before dosing and then resumed feeding 4 hours after dosing. All animals received an intramuscular injection of 10 μg / kg pentagastrin 30 minutes before dosing.

[0318] Sampling time: 0.5h, 1h, 1.5h, 2h, 3h, 4h, 6h, 8h, 12h, 16h and 24h after the end of administration.

[0319] Anticoagulant: EDTA-2K anticoagulant; centrifuge at 4500 rpm for 10 minutes to separate plasma samples, and store in a -70°C refrigerator.

[0320] Biological sample analysis method: The concentration of ibrutinib in plasma samples was detected by tandem mass spectrometry (LC-MS\MS).

[0321] Phoenix WinNonlin 7.0 was used to calculate the pharmacokinetic parameters (AUC 0-t , Cmax, Tmax and the arithmetic mean (± SD) and geometric mean of these parameters. The results are shown in Table 15 below.

[0322] Table 15. Oral pharmacokinetics of different formulations of ibrutinib in beagle dogs (n=3)

[0323] The results of the beagle dog PK experiment showed that IRB-F7 and IRB-F8 had significantly lower Cmax and AUC than RLD+NR. 0-lastThe differences were significant (P < 0.01 and 0.001). NR is a compound reported in the literature to inhibit CYP3A4 or P-gp. The relative bioavailability of IRB-F7 and IRB-F8 supramolecular self-assembled complexes directly encapsulated compared to the reference formulation plus NR was 2.9-fold and 7.3-fold, respectively. The IRB-F7 and IRB-F8 supramolecular self-assembled complexes differ in the carriers used to control zeta potential. The IRB-F7 formulation uses RBDS-B-Na as the zeta potential carrier, while the IRB-F8 formulation uses STVB-Arga as the zeta potential carrier. The resulting supramolecular self-assembled complexes have zeta potentials of -10.8 mV and +9.6 mV, respectively. The latter is more likely to penetrate the intestinal mucus layer and intestinal epithelial cells. Therefore, when the target delivery agent is formulated into a supramolecular self-assembled complex, not only does it significantly improve enzyme metabolic stability, but it also significantly enhances gastrointestinal absorption efficiency.

[0324] Example 8. Preparation of supramolecular self-assembled complexes containing paclitaxel (PTX) and docetaxel (DTX)

[0325] Paclitaxel, docetaxel, cephalomannine, and cabazitaxel are tumor microtubule inhibitors clinically used to treat a variety of solid tumors. Paclitaxel and docetaxel are representative model drugs for validation, and the technical solutions used for paclitaxel and docetaxel are also applicable to cephalomannine and cabazitaxel.

[0326] The formula of the supramolecular self-assembled complex of paclitaxel and docetaxel is shown in Table 16 below:

[0327] Table 16. Design of supramolecular self-assembled complex formulations of paclitaxel and docetaxel

[0328] (1) Preparation process

[0329] Method I: Solvent Injection

[0330] ① Dissolve PTX in THF at a concentration of 250 mg / ml.

[0331] ② Dissolve the carrier and / or high molecular weight polymer in 5 ml of water or pH 7.4 phosphate buffer;

[0332] While stirring, slowly add ① to ② until obvious turbidity appears. Continue stirring for 30 minutes. Concentrate on a rotary evaporator at ≤45℃ until there is no solvent smell. Centrifuge at 15000rpm for 5 minutes.

[0333] Method II: Thin Film Dispersion Method

[0334] The carrier, high molecular weight polymer, PTX or DTX were dissolved in ethanol-water solution (10 ml EtOH + 1 ml water), the solvent was recovered under reduced pressure at ≤45°C, 5 ml water was added, the mixture was shaken at 250 rpm at 37°C for 2 hours, and centrifuged at 15,000 rpm for 5 minutes.

[0335] Method III: One-step method

[0336] The vector, PTX or DTX were dissolved in an ethanol-water solution (15 ml EtOH + 5.5 ml water), and the ethanol was recovered under reduced pressure until there was no alcohol smell, and the solution was centrifuged at 15000 rpm for 5 min.

[0337] (2) Content determination, solidification and re-dissolution results

[0338] The content of paclitaxel and docetaxel supramolecular self-assembled complexes was determined using the high performance liquid chromatography method shown in Table 1. The formulation with the highest content was freeze-dried, and the encapsulation efficiency and drug loading were calculated. A reconstitution experiment was conducted, and the particle size and Zera potential of the reconstituted solution were measured. The results are shown in Table 17 below:

[0339] Table 17. Evaluation of properties of paclitaxel / docetaxel supramolecular self-assembled complex

[0340] The above results show that when single carriers, RBDS-A (F15) or MGSD-V (F23), were supramolecularly self-assembled with paclitaxel at a mass ratio of 15:1, the encapsulation efficiency was only 36.1% and 18.1% respectively. When RBDS-A was combined with RBDS-M (F16), the carrier and paclitaxel were assembled at a mass ratio of 15:1 using the same preparation process, and the encapsulation efficiency was significantly improved to 64.5%. When RBDS-A and RBDS-M were combined and then used with the polymer Soluplu or PVP K30, respectively, the carrier and paclitaxel mass ratio was 3:1 and the polymer and paclitaxel mass ratio was 4:1, and the same preparation process was used for assembly, the encapsulation efficiency was 72.4% and 48.5%, respectively. The encapsulation efficiency was higher when the carrier was combined with Soluplus. RBDS-B was combined with RBDS-M, STVS, and MGSD-V, and then with Soluplus, to prepare supramolecular self-assembled complexes using the I / III / III method (F20, F21, and F22). The encapsulation efficiencies were 22.9%, 93.7%, and 100.4%, respectively. The resulting supramolecular self-assembled complex solutions contained paclitaxel at concentrations of 6.86 mg / ml, 37.22 mg / ml, and 65.16 mg / ml, respectively. The drug loadings for each formulation were 3.78%, 20.1%, and 35.8%, respectively. The encapsulation efficiency and drug loading of F22, containing the charge carrier RBDS-B-His, were significantly higher than those of F21, prepared using the same method. Formulations F17, F21, and F22, solidified by freeze-drying and then reconstituted with purified water, remained clear upon addition of 50 ml of water. The reconstituted solutions had a narrow hydrated particle size distribution, with an average particle size of less than 100 nm. When the surfactants SDS or TPGS were added to the F21 formulation and prepared using either Method II or Method III, respectively, the encapsulation efficiencies of F26 and F27 were significantly lower (approximately 62.5% and 65.3%) compared to F21, and the drug loadings were 9.82% and 10.50%, respectively, both significantly lower than F21 (encapsulation efficiency 93.7% and drug loading 20.1%). The addition of the surfactants SDS or TPGS to the formulation did not exert a synergistic effect. F25 and F26, prepared using the same formulation containing the surfactant SDS using Method III and Method II, achieved encapsulation efficiencies of 6.02% and 62.5%, respectively, yielding significantly different results.

[0341] The combined carrier (RBDS-A + RBDS-M + STVS + RBDS-C) and docetaxel mass ratio was 15:8, and the polymer to docetaxel mass ratio was 5:2. Taurine, a pH adjuster, was added to each formulation. Supramolecular self-assembled complexes were prepared using Method II (F29) and Method III (F30), respectively. The encapsulation efficiencies of the two formulations were 23.9% and 97.5%, respectively, and the drug loading capacity of F30 was 21.9%. F31, based on F30, added the carrier MGSD-V, while keeping all other components unchanged. F32, based on F31 but omitting the taurine, was prepared using Method III. The encapsulation efficiencies of the two formulations were 96.3% and 101.4%, respectively, and the drug loading capacities were 25.6% and 38.6%, respectively, significantly improving both the encapsulation efficiency and drug loading capacity. Formulations F31 and F32 were both clear upon solidification and reconstitution. Formulas F33 and F34 are combinations of RBDS-A and RBDS-M, respectively, and then used in combination with Soluplus or PVP VA64, respectively. The supramolecular self-assembly complexes were prepared using method II. The encapsulation efficiencies of the two formulas were 79.3% and 26.0%, respectively, and formula F33 can be redissolved after solidification.

[0342] (3) Transmembrane permeability evaluation

[0343] Paclitaxel and docetaxel are substrates of the cytochrome P450 subunits CYP2C8 and CYP3A4, as well as the efflux protein P-gp. Paclitaxel has an oral bioavailability of less than 2%. Therefore, current systemic therapy with paclitaxel and docetaxel is still primarily administered by injection. To achieve effective oral administration of paclitaxel and docetaxel, two key bottlenecks must be overcome: permeability across intestinal epithelial cells and enzymatic degradation.

[0344] The transmembrane permeability of the supramolecular self-assembled complex was evaluated using the rat everted intestinal sac (EGS) method. SD rats were fasted for 12 hours prior to the experiment and had free access to water. After sacrifice by cervical dislocation, the abdominal cavity was opened and approximately 10 cm of the jejunum was quickly removed. After rapid flushing with cold Krebs-Ringer solution, the jejunum was inverted with a glass rod and ligated with surgical suture to form an intestinal sac. The jejunal sacs were fixed and each sac was injected with 2 mL of Krebs-Ringer solution using a syringe. The intestinal segments were then placed in a beaker containing 500 mL of Krebs-Ringer solution containing a fixed concentration of paclitaxel or docetaxel at 37°C. The content of the supramolecular self-assembled complexes was calculated based on the measured content. Paclitaxel supramolecular compositions were formulated as F15-F17 (added as aqueous solution, excluding water introduced by the solution), F21, and F22 (added as powder), while docetaxel compositions were formulated as F31, F32, and F33 (added as powder). The initial concentration of both paclitaxel and docetaxel was 50 μg / mL. Stirring was initiated, and a CO₂-O₂ (5:95) gas mixture was introduced into the medium during incubation. The sacs were then placed into the incubation compartments according to their respective groups. At 30, 60, 90, and 120 minutes, 200 μL of the internal fluid from each intestinal sac was aspirated as the test solution, and the same volume of Krebs-Ringer solution incubated at 37°C was added. At the end of the experiment, the intestinal sac area was measured. The protein of the test solution was precipitated with 3 volumes of acetonitrile, and the solution was centrifuged at 4500 rpm and 37°C for 10 minutes. 1 mL of the supernatant was collected and nitrogen-blown to 0.5 mL. The supernatant was then analyzed by high-performance liquid chromatography for paclitaxel or docetaxel concentration. The paclitaxel and docetaxel contents were calculated using the external standard method. At the end of the experiment, the length and width of the intestinal segment were measured after dissection, and the permeation area was calculated.

[0345] According to the permeation drug concentration measured at each sampling point, the cumulative permeation amount of drug Q (ng), drug permeation rate constant K and apparent permeability coefficient (P app， cm·s -1 ), the results are shown in Table 18 below:

[0346] Table 18. Apparent permeability coefficient investigation by everted intestinal sac method (n=3)

[0347] Results from the everted intestinal sac assay indicate that the transintestinal mucosal permeability of paclitaxel or docetaxel formulations F16, F17, F20, F21, F-22, and F31-F33 is significantly superior to that of formulation F15 (RBDS-A). The multi-carrier combination formulation with different chemical structures is particularly superior. Formulations F-21 and F-22 are essentially identical in composition, differing in that F-21 contains RBDS-B sodium salt (negatively charged) while F-22 contains RBDS-B-His conjugate (positively charged). The permeability of the supramolecular composition F-22 is higher than that of F-21 and the other formulations, as shown in Figure 14. Particle size measurements of paclitaxel formulations F22 and docetaxel formulations 32, diluted 200-fold, are shown in Figures 15-16.

[0348] (4) Pharmacokinetic study of paclitaxel supramolecular self-assembled complex in rats

[0349] Experimental Animals: Male SD rats weighing 250 g ± 30 g were housed in the same environment for one week prior to the experiment. Rats were fasted for at least 10 hours prior to dosing, with or without water for two hours before dosing. Following dosing, rats were allowed free access to water for two hours and food for four hours. The animals were divided into four groups, each containing five rats.

[0350] Paclitaxel supramolecular self-assembled complexes PTX-F17, PTX-F21 and PTX-F22 were administered at 30 mg / kg. All supramolecular self-assembled complexes were dissolved in distilled water to form 1 ml solution and directly administered orally.

[0351] Preparation of Taxol: Pass paclitaxel through a 100-mesh sieve, dissolve it in polyoxyethylene castor oil-anhydrous ethanol (1:1, V / V) to a 6 mg / ml solution, and filter.

[0352] Sampling time: 0.5h, 1h, 1.5h, 2h, 2.5h, 3h, 4h, 6h, 8h, 12h and 24h; under isoflurane anesthesia, about 0.4ml of blood was collected from the inner canthus, sodium heparin was used as an anticoagulant, and the blood was centrifuged at 4500rpm for 10 minutes, and 100μl of plasma was collected as the test sample.

[0353] Each prescription was administered to 5 rats in total. The administration method was direct oral administration of 1.3 ml of Taxol injection solution and 1 ml of supramolecular self-assembled complex. All animals were given 1 ml of water after administration.

[0354] Parameters of the tandem mass spectrometry (LC-MS\MS) method for the determination of paclitaxel plasma sample concentrations: the chromatographic column was a Waters Synergi RP column with specifications of 4 μm, 2.0 × 50 mm, USA;

[0355] Mobile phase A: 0.1% (v / v) formic acid-water solution; mobile phase B: 0.1% (v / v) formic acid-acetonitrile solution; flow rate: 0.3 mL / min; injection volume: 5 μL; column temperature: 25°C;

[0356] The gradient elution program is shown in Table 19 below:

[0357] The mass spectrometry quantification mode of paclitaxel was: MRM positive ion mode, m / z 854.5→286.0, ion source voltage: 5000V; collision energy: 20V, ion source temperature: 500℃; the mass spectrometry quantification mode of internal standard diphenhydramine was: SIR positive ion mode, m / z 256.3, ion source voltage: 5000V; ion source temperature: 500℃;

[0358] Plasma sample processing method: Accurately measure 100 μl of plasma sample and place it in a 1.5 ml centrifuge tube. Add 10 μl of internal standard diphenhydramine solution (concentration 200 ng / ml, 50% acetonitrile), add 500 μl of methyl tert-butyl ether solvent protein precipitation, vortex for 2 minutes to mix thoroughly, centrifuge the sample at room temperature at 12000 × g for 5 minutes, and then place the sample in a -80°C refrigerator for 25 minutes. Transfer the supernatant to a clean glass tube, blow dry with nitrogen (nitrogen temperature 37°C), and then reconstitute with 100 μl of acetonitrile: water: formic acid (50:50:0.1, v / v / v) solution and vortex for 1 minute; aspirate 5 μl of supernatant and inject it into the LC-MS / MS system for analysis;

[0359] Phoenix WinNonlin 7.0 was used to calculate the pharmacokinetic parameters (AUC 0-t , Cmax, Tmax and the arithmetic mean (± SD) of these parameters). The results are shown in Table 20 below.

[0360] Table 20. Oral pharmacokinetic results of paclitaxel supramolecular self-assembled complex in rats (n=5)

[0361] Oral administration of the supramolecular self-assembled complexes in rats revealed that, compared with oral administration of Taxol solution, peak plasma drug levels and drug exposure for each formulation were 4.8 to 9.0 times and 9.5 to 20.1 times higher, respectively, than those observed with Taxol. Drug absorption permeability and oral bioavailability were significantly improved, and the elimination half-life of PTX-F21 and PTX-F22 was significantly prolonged by approximately one-fold. However, the oral bioavailability of PTX-F22 was significantly superior to that of PTX-F17 and PTX-F21, and far superior to that of Taxol. These results are generally consistent with those obtained with the everted intestinal sac method.

[0362] Example 9. Preparation of supramolecular self-assembled complex of lenvatinib and apixaban

[0363] (1) The formula is designed as follows:

[0364] Table 21. Design of formulation composition of each supramolecular self-assembly complex

[0365] (2) Preparation method

[0366] According to the formulas F35, F36, F42 and F43 in Table 21, the supramolecular self-assembled complexes of lenvatinib and apixaban were prepared respectively by a one-step method.

[0367] 1) Prepare 40 ml of a pH 6.8 phosphate buffer-acetonitrile (1:3, v / v) solution, add the carrier, then the polymer, adjust the pH to 6.8-7.4 with Tris, then add 0.2 g of apixaban and stir to mix thoroughly. Ultrasonicate at 45-50°C to dissolve completely. Add 6 ml of pH 6.8 phosphate buffer and continue stirring for 1 hour. Concentrate on a rotary evaporator at 45°C until no organic solvent odor is present. Let the aqueous solution stand, centrifuge at 15,000 rpm, and collect the supernatant. Formulations F35 and F36 contain more residual solids.

[0368] 2) Prepare a pH 6.8 phosphate buffer-methanol (1:3, v / v) solution, add the carrier, then the polymer, adjust the pH to 6.8-7.4 with Tris, then add 0.2 g of lenvatinib to the system, sonicate at 55°C, then add 15 ml of tetrahydrofuran until completely dissolved. Continue stirring for 1 hour, and concentrate on a rotary evaporator at 45°C until no organic solvent odor is present. Let the aqueous solution stand, centrifuge at 15,000 rpm, and remove the supernatant. F35, F36, and F43 retain a significant amount of solids.

[0369] The content was detected by high performance liquid chromatography, and the encapsulation efficiency was calculated according to the dosage and the measured content.

[0370] (3) Evaluation of the properties of supramolecular self-assembled complexes of lenvatinib and apixaban

[0371] Table 22. Evaluation of properties of supramolecular self-assembled complexes of apixaban and lenvatinib

[0372] Based on the above results, NRGDC is solubility-limited, and the carrier dosage should generally not exceed 0.2% at room temperature; otherwise, significant precipitation will occur. Formulations F35 and F36, using NRGDC or MGSD-V alone as the carrier, with active ingredient to carrier mass ratios of 2:5 or 1:5, respectively, showed poor self-assembly of the active ingredient, resulting in significant solid precipitation after centrifugation. Formulations F42 and F43, combining NRGDC or NHDC with MGSD-V, respectively, and then incorporating them into the high-molecular-weight polymer HPMCAS, significantly improved the carrier's self-assembly ability, resulting in significantly higher encapsulation efficiency and drug loading.

[0373] Apixaban-F42, Apixaban-F43, Lenvatinib-F42, and Lenvatinib-F43 were sealed and stored at room temperature (15°C-20°C) for one week without precipitation.

[0374] (4) Investigation of the dissolution rate of apixaban supramolecular self-assembled complex

[0375] Using apixaban N-1 crystal form (WO2011106478A2) as a control, the N-1 crystal form API was micronized before dissolution, and D90 was controlled to be ≤73 μm.

[0376] Dissolution method: Paddle method, 100 rpm, 37°C. Sampling points: 15, 30, 60, and 480 min.

[0377] Dissolution medium: pH 6.8 phosphate buffer, medium volume 250 ml.

[0378] Apixaban-F42 and F43 supramolecular self-assembled complexes were sampled at 25 mg of the target delivery substance, and the crystalline drug was directly sampled to investigate the dissolution rate.

[0379] 5 ml of the sample was collected from each sampling point and centrifuged at 37°C and 13,000 rpm. 0.5 ml of the supernatant was collected from the drug substance group and diluted to 1 ml with 50% acetonitrile. 0.2 ml of the supernatant was collected from the supramolecular composition group and diluted to 1 ml with 50% acetonitrile. The apixaban content was determined by HPLC, and the solubility at each sampling point was calculated and converted into concentration (μg / ml).

[0380] Three cups were prepared for each formulation, and the sample volume in each dissolution cup was converted to 25 mg of apixaban. The dissolution curve is shown in Figure 17.

[0381] Table 23. Comparison of dissolution rates of supramolecular self-assembled complexes of Apixaban and N-1 crystal form (n=3)

[0382] Based on the above results, the cumulative dissolution rates of apixaban and carrier-based supramolecular self-assembled complexes F42 and F43 exceeded 80% after 30 minutes, and were 94.2% and 97.7% after 60 minutes, respectively, indicating near-complete dissolution. The dissolution rates of the same dose of apixaban Form N-1 were 7.9% and 16.0% after 30 and 60 minutes, respectively, with a slow increase after 480 minutes. The dissolution rates of each supramolecular self-assembled complex at 15 minutes were approximately 10-11 times higher than those of the micronized Form N-1, significantly improving the dissolution rate of apixaban. See Table 24 below for details.

[0383] Table 24. Calculation of dissolution rate of supramolecular self-assembled complex of Apixaban Note 1: The ratio of the cumulative dissolution rate of the supramolecular self-assembled complex to the dissolution rate of the N-1 crystal form at 15 minutes.

[0384] (5) Stability study of lenvatinib supramolecular self-assembled complex

[0385] The formulations of Lenvatinib-F42 and Lenvatinib-F43 were freeze-dried for 48 hours, respectively, and packaged in double aluminum packages. Samples were placed at a high temperature of 60°C ± 2°C, and samples were taken on the 10th and 30th days to detect related substances and genotoxic impurities. The effect of high temperature on the stability of the target delivery product was investigated. The comparison of the test results of the influencing factors of the formulation of the present invention and the reference formulation Lenvima at 30 days and 0 days under the same conditions is shown in Table 25 below:

[0386] Table 25. Changes in related substances and genotoxic impurities of lenvatinib supramolecular self-assembled complex at high temperature of 60℃ for 30 days

[0387] From the results of the influencing factors of high temperature of 60℃, it can be seen that the genotoxic impurities ZZ-D and ZZ-C of the supramolecular self-assembled complex increased slightly after being placed at high temperature of 60℃ for 30 days. The growth rate of ZZ-C impurity in F43 formula was slightly faster than that in F42 formula, but ZZ-C and ZZ-D of the two formulas did not exceed the control limit of 400ppm after being placed at high temperature of 60℃ for 30 days; the common degradation impurity F of the two formulas was detected after being placed at high temperature of 60℃ for 30 days, but the detection amount was very low. The chemical stability of the genotoxic impurities and common degradation impurities of the supramolecular self-assembled complexes prepared by F42 and F43 formulas after being placed at high temperature of 60℃ for 30 days was not inferior to that of the reference preparation Lenvima.

[0388] Example 10: Preparation of supramolecular self-assembled complexes of nilotinib, nintedanib, dabigatran etexilate, and olaparib

[0389] The supramolecular self-assembled complexes of nilotinib, nintedanib, dabigatran etexilate and olaparib were prepared respectively using the formulations shown in Table 26.

[0390] Table 26. Formulations of supramolecular self-assembled complexes of nilotinib, nintedanib, dabigatran etexilate, and olaparib

[0391] (1) Preparation process

[0392] 1) Preparation of target delivery solution

[0393] Weigh 1g of nintedanib and dissolve it in 100g of methanol;

[0394] Weigh 1g of nilotinib or olaparib, dissolve in 100ml of dichloromethane-methanol (3:1, v / v), and set aside;

[0395] Weigh 1 g of dabigatran etexilate and dissolve it in 20 ml of ethanol-dichloromethane (2:1, V / V) to 50 mg / ml for later use.

[0396] 2) Add 10 ml of water to the carrier and polymer. If insoluble, heat to 60°C. Once dissolved, add the drug-containing organic solvent from step 1) dropwise to the aqueous phase while stirring until noticeable turbidity develops. Stop adding dropwise and continue stirring for 60 minutes. Concentrate on a rotary evaporator until the solvent odor is eliminated. Allow to stand, centrifuge at 15,000 rpm for 5 minutes, and separate the supernatant. If no turbidity, stratification, or precipitation is observed after standing at room temperature for 48 hours, evaluate the properties.

[0397] (2) Characteristic evaluation

[0398] The content of each active ingredient in Table 1 was detected by high performance liquid chromatography, and the encapsulation efficiency and drug loading were calculated. The particle size and zeta potential of the formulation with high drug loading were measured. The results are shown in Table 27 below.

[0399] Table 27. Evaluation of properties of supramolecular self-assembled complexes of nintedanib, nilotinib, and olaparib

[0400] (3) Study on oral bioavailability of nintedanib supramolecular self-assembled complex in rats

[0401] Experimental animals: 21 Wistar male rats, weighing 220 g ± 30 g, fasted for more than 12 hours before administration and had free access to water.

[0402] Rats were randomly divided into three groups, each with seven rats. Each group was administered a reference formulation (50 mg / kg of softgel contents), NDNB-F44, or NDNB-F46 formulation. The oral dose was calculated based on the actual content, resulting in a 20 mg / kg dose. Each group was then given 1 ml of purified water. Water was allowed ad libitum for 2 hours and food for 4 hours after administration. Four hours after administration, two rats from each group were anesthetized with ether and rapidly dissected. The livers were isolated, rinsed with cold saline, blotted dry with absorbent paper, and weighed.

[0403] 0.5 ml of blood was collected from the inner canthus at 0.5 h, 1 h, 1.5 h, 2 h, 3 h, 4 h, 6 h, 8 h, and 10 h after administration, and anticoagulated with sodium heparin (0.3%). After blood collection, the blood was centrifuged at 2800 g for 10 minutes (2-8°C), and 150 μl of plasma was separated for detection. The samples were stored in a -80°C refrigerator before detection.

[0404] Plasma samples were tested by LC / MS / MS with ibrutinib as the internal standard. Pharmacokinetic parameters were calculated using Phoenix WinNonlin 7.0 based on the plasma drug concentration data at different time points, providing AUC 0-t , AUC 0-inf 、C max 、T max and T 1 / 2 The relative bioavailability was calculated based on the parameters and their mean and standard deviation. The results are shown in Table 28 below:

[0405] 1) PK parameters of nintedanib supramolecular self-assembled complex and reference preparation

[0406] Table 28. Comparison of oral PK of nintedanib supramolecular self-assembled complex and reference formulation in rats

[0407] In summary, when nintedanib was formulated into supramolecular self-assembling complexes and administered orally to rats, the relative bioavailabilities of NDNB-F44 and NDNB-F46 were 1706% and 1900%, respectively, compared to the reference formulation (Vitaga), representing 17 and 19 times the relative bioavailability of the reference formulation in humans. The reference formulation's oral bioavailability in humans was only 4.7%. The PK profiles of the nintedanib supramolecular complexes showed two peaks for both formulations, while the reference formulation had a single peak. PK studies in rats demonstrated that nintedanib formulated into supramolecular self-assembling complexes can achieve controlled release with a once-daily dosage.

[0408] 2) Liver tissue distribution

[0409] Rat liver tissue was taken and quickly minced. Cold physiological saline was added to a mortar at a ratio of tissue (g): physiological saline (g) = 1:4. The tissue was quickly ground into a homogenous state under low temperature and dark conditions. 140ul of liver tissue homogenate was taken, 10ul of internal standard solution was added, vortexed for 10s, 900ul of MeOH solvent was added, vortexed for 30s, 140ul of H2O was added, vortexed for 10s, and allowed to stand at 4°C for 10min. Centrifuged (1500rpm, 4°C, 10min), and the supernatant was analyzed by liquid chromatography-mass spectrometry to detect nintedanib and its metabolite BIBF1202. The results are shown in Figure 18:

[0410] As shown in Figure 18, the levels of the prototype nintedanib drug and its lipid metabolite BIBF1202 measured in the rat liver 4 hours after oral administration of the supramolecular self-assembled complex formulation were significantly lower (10 times) than those of the reference formulation, indicating that the two supramolecular self-assembled complex formulations may be mainly absorbed through the lymphatic system, thereby reducing the first-pass metabolism in the liver, thereby significantly reducing the toxicity of nintedanib to the liver.

[0411] (4) Study on esterase metabolism of dabigatran etexilate and nintedanib supramolecular self-assembled complex

[0412] Dabigatran etexilate and nintedanib were used as target delivery agents, curcumin was used as an inhibitor of esterase, and bis-p-nitrophenyl phosphate (BNPP) was used as a positive inhibitor. The experiment was carried out according to the following steps.

[0413] Except for rhesus monkeys or cynomolgus monkeys, rat esterase has a high homology with human, so ileal intestinal fluid of SD rats was collected for esterase metabolism evaluation.

[0414] 1) Preparation of rat ileal intestinal juice

[0415] Male SD rats (weighing 250g-300g) were fasted for at least 16 hours with free access to water. The next day, rats were gavaged with 2ml of water and then fed. 30 minutes after feeding, rats were anesthetized with ether and sacrificed. The abdomen was opened along the midline of the abdomen, and approximately 20cm of fresh ileum was immediately removed. The ileal contents were collected (operated on ice) and centrifuged (4000g, 4°C, 10 minutes). The supernatant was collected as rat ileal fluid. The supernatant was placed on ice until needed or stored in a -80°C refrigerator.

[0416] 2) Preparation of rat intestinal tissue perfusion fluid

[0417] The intestinal tissue was perfused with 4°C Krebs phosphate buffer (pH 7.4), both ends of the intestinal tissue were tied tightly, and the tissue was placed in a 4°C 0.9% NaCl solution for 20 min. The intestinal perfusion fluid was collected into a centrifuge tube and centrifuged (centrifugal force 4000g, 4°C, 10 min). The supernatant was taken as the rat ileum perfusion fluid and placed on ice for later use, or stored in a -80°C refrigerator.

[0418] The positive inhibitor BNPP was prepared into a 100 μM solution using Krebs solution, in which curcumin needed to be solubilized with a small amount of DMSO.

[0419] Take an appropriate amount of nintedanib or dabigatran etexilate free base, dissolve it in dimethyl sulfoxide and quantitatively dilute it to prepare a solution containing approximately 250 μM of nintedanib or dabigatran etexilate per 1 ml, which is used as the stock solution of nintedanib or dabigatran etexilate, respectively.

[0420] According to the measured content of dabigatran etexilate-F47 and nintedanib-F47 supramolecular self-assembled complexes, a certain volume of supramolecular self-assembled complex solution was taken and diluted with pH 7.4 Krebs solution to prepare a solution containing approximately 500 μM nintedanib or dabigatran etexilate per 1 ml, which was used as the test solution.

[0421] The positive control group contained only the target delivery substance and 70% ileal intestinal fluid; the negative control contained only the target delivery substance and Krebs solution;

[0422] Before the experiment, rat ileal intestinal juice was diluted 70% with Krebs solution and recorded as 70% ileal intestinal juice.

[0423] Table 29. Experimental design (n=3)

[0424] The specific operation is as follows: Krebs solution and 70% ileal intestinal juice are preheated at 37°C for 10 minutes, and after adding the target delivery drug stock solution according to the above scheme, vortex for 10 seconds; start timing, sample 100 μl at 0.5h, 1h and 3h, add 300 μl of methanol (nintedanib) or ethanol (dabigatran etexilate), vortex for 30 seconds, let it stand for 10 minutes, centrifuge (13000 rpm, 4°C, 10 minutes), take the supernatant for detection, calculate the amount of nintedanib and dabigatran etexilate prototype drugs, perform offline regression based on the measured drug concentrations at each sampling point, obtain the elimination rate equation and correlation coefficient, calculate the elimination rate constant and half-life, and the results are as follows:

[0425] Table 30. Esterase metabolism test results

[0426] From the above results, it can be seen that: the degradation amount of the negative control group of nintedanib and dabigatran etexilate was less than 1% when incubated at 37°C for 4 hours, which is considered to be the thermodynamic temperature; the positive control group of dabigatran etexilate and nintedanib was significantly degraded, with elimination rate constants of 0.42 and 0.39, respectively, and elimination half-lives of 1.65h and 1.79h, respectively; after the addition of the positive inhibitor BNPP, the esterase activity was significantly inhibited, and the elimination half-lives of dabigatran etexilate and nintedanib were extended to 8.50h and 6.84h, respectively; while the elimination half-lives of the dabigatran etexilate and nintedanib supramolecular self-assembled complex groups were 7.92h and 6.94h, respectively, and the esterase metabolic stability was significantly improved.

[0427] Similar drugs or prodrugs also include, but are not limited to, the following drugs: aspirin, tenofovir alafenamide, capecitabine, cilazapril, clopidogrel, cocaine, enalapril, heroin, imidapril, irinotecan, pethidine, methylphenidate, olmesartan, orlistat, oseltamivir, quinapril, ramipril, temocapril, trandolapril, benazepril, etc. are also applicable.

[0428] Example 11. Cannabidiol and Ticagrelor Supramolecular Self-Assembled Complex

[0429] (1) Preparation of supramolecular self-assembly complexes

[0430] Supramolecular compositions of cannabidiol and ticagrelor were prepared according to the formula in Table 31:

[0431] Dabigatran drug-containing solution: add 3g of dabigatran etexilate to 10ml of dichloromethane-ethanol (3:1, V / V) mixed solvent, sonicate to completely dissolve, and set aside.

[0432] Cannabidiol-containing solution: Dissolve cannabidiol in ethanol to form a solution containing approximately 300 mg of cannabidiol per 1 ml, and set aside;

[0433] Ticagrelor solution: Dissolve ticagrelor in methanol to form a solution containing 300 mg of ticagrelor per 1 ml, and set aside.

[0434] Prepare a pH 6.8 phosphate buffer solution and add the carrier and surfactant separately. If insoluble, heat to 50°C to dissolve. Take 10 ml of each solution. For F37, adjust the pH to 6.8 with an appropriate amount of Tris or taurine; for the other formulations, adjust the pH to 6.5 with Tris or taurine. While stirring, slowly add the drug-containing organic phase dropwise until noticeable turbidity appears. Stop adding and continue stirring until the organic solvent smell disappears. Let the solution stand and centrifuge at 13,000 rpm for 5 minutes to separate the supernatant.

[0435] (2) Evaluation of supramolecular composite properties

[0436] The above solutions were kept sealed at room temperature for 72 hours and observed for demixing, precipitation, or precipitation. Solutions that did not undergo phase separation were sampled for content analysis, freeze-dried to solidify, and reconstituted in 100 ml of pH 7.4 phosphate buffer, adding small amounts repeatedly. The reconstituted supramolecular compositions were tested for particle size distribution and zeta potential. The results are shown in Table 32 below.

[0437] Table 32. Evaluation of properties of supramolecular compositions of cannabidiol and ticagrelor

[0438] In summary, the supramolecular compositions prepared by the above formulations are easy to redissolve after solidification. After redissolution, the particle size distribution is all below 100 nm, indicating a uniform particle size distribution.

[0439] (3) Investigation of the dissolution rate of ticagrelor supramolecular composition

[0440] After the ticagrelor raw material was micronized, the D90 was measured by a laser particle size analyzer wet method and was 11.3 μm.

[0441] Paddle method was used, 100 rpm, 37°C.

[0442] Dissolution medium: pH 6.8 phosphate buffer, medium volume: 250 mL;

[0443] Sampling points: 15, 30, 60, 120, 180, 480 min.

[0444] The dissolution experiment of the supramolecular composition of ticagrelor was carried out by converting the measured content into a unit dose of 60 mg;

[0445] Sample pretreatment: 5 ml of the sample was centrifuged at 37°C and 13,000 rpm, 0.2 ml of the supernatant was taken and diluted to 1 ml with 70% methanol; the content of ticagrelor was detected by HPLC, and the solubility (%) of each sampling point was calculated.

[0446] Table 33. Comparison of dissolution rates of ticagrelor supramolecular composition and ticagrelor micronized bulk drug

[0447] According to the above results, only 28.3% of the micronized API was dissolved in 480 min. In 15 min, the dissolution rates of the supramolecular compositions F37, F40, and F41 relative to the API were 9.3 times, 3.6 times, and 8.8 times that of the API, respectively. The dissolution rate was significantly improved, among which the formulation containing RBDS-B carrier had the fastest dissolution rate.

[0448] (4) Oral pharmacokinetics study of supramolecular compositions in rats

[0449] The cannabidiol supramolecular compositions F-37 and F-40 were directly encapsulated and orally administered to rats. The cannabidiol API was also encapsulated as a control.

[0450] Experimental Animals: Male SD rats weighing 250 g ± 30 g were used and housed in the same environment for one week before the experiment. Rats were fasted for at least 10 hours before administration, with or without water for two hours before administration and free access to water for two hours after administration.

[0451] Experimental drugs: F-37C capsules, F-40C capsules, cannabidiol capsules, dosage: 50 mg / kg body weight

[0452] Sampling points: 0.25h, 0.5h, 1h, 1.5h, 2h, 4h, 6h, 8h, 12h and 24h. Under isoflurane anesthesia, about 0.5ml of blood was collected from the inner canthus. Sodium heparin was used as an anticoagulant. The blood was centrifuged at 3500rpm for 10 minutes, and 150μl of plasma was collected as the test sample.

[0453] Each formulation was administered to 5 rats in total. The administration method was capsule administration using a rat administration device, and 2 ml of water was administered orally after administration.

[0454] Analytical method: UPLC-tandem mass spectrometry (UPLC-MS\MS) was used. The chromatographic conditions were as follows:

[0455] The chromatographic column is Waters (5 μm, 2.1 × 10 cm, Waters USA); mobile phase A: 1 mM ammonium formate; mobile phase B: acetonitrile; flow rate: 0.3 mL / min; injection volume: 10 μL; column temperature: 35°C;

[0456] Gradient elution, the gradient elution program is as shown in Table 34:

[0457] Cannabidiol quantification mode: MRM negative ion mode, 313.085→245.033; ion source voltage: -4500V; collision energy: -28V, ion source temperature: 400℃; internal standard clobetasol propionate mass spectrometry quantification mode: MRM positive ion mode, 467.161→372.900, ion source voltage: 5500V; collision energy: 13; ion source temperature: 400℃;

[0458] The method LLOQ was 5 ng / mL.

[0459] Phoenix WinNonlin 7.0 was used to calculate the pharmacokinetic parameters (AUC 0-t , Cmax, Tmax and the arithmetic mean (± SD) of these parameters). The results are shown in Table 35 below.

[0460] Table 35. Oral pharmacokinetic results of cannabidiol supramolecular composition in rats

[0461] Cmax and AUC of F-37C and F-40C 0-t The ratios of the corresponding values ​​of API were 6.9 and 5.9 times, respectively, which were significantly different. The oral relative bioavailability of formulations F-37C and F-40C was expressed as AUC 0-inf The calculated results are 7.5 times and 6.1 times that of the API respectively; the elimination half-life is 2.2 times and 1.9 times that of the API respectively, and the in vivo elimination half-life is significantly prolonged.

[0462] Example 12: Celecoxib, MRTX1133 supramolecular composition

[0463] (1) Preparation of supramolecular compositions

[0464] Celecoxib and MRTX1133 supramolecular compositions were prepared according to the formulas in Table 36.

[0465] Celecoxib was prepared into a 250 mg / ml solution with anhydrous ethanol and set aside;

[0466] MRT1133 was dissolved in acetone-dichloromethane (2:1, V / V) to a solution containing approximately 30 mg per 1 ml as the organic phase and protected from light.

[0467] Take each carrier and polymer, add 10 ml of water, sonicate for 10 minutes to dissolve, and adjust the pH to approximately 6.8. While stirring, slowly add the drug-containing organic phase dropwise to the aqueous phase until noticeable turbidity appears. Stop the addition and continue stirring for 30-60 minutes. Concentrate on a rotary evaporator until there is no solvent odor. Allow to stand, centrifuge, and collect the supernatant. Allow to stand at room temperature for 72 hours. If there is no phase separation, proceed to subsequent evaluation.

[0468] (2) Evaluation of supramolecular composite properties

[0469] Table 37. Evaluation of properties of supramolecular compositions of celecoxib and MRTX1133

[0470] Formula F49,50 simulates in situ assembly of the digestive tract in vivo. After oral administration, the supramolecular composition enters the small intestine through the stomach and duodenum. The β-glucosidase in the small intestine acts on the β-glycosidic bonds on the carrier, gradually enzymatically hydrolyzing the glucose residues on the carrier. The specific process is as follows:

[0471] The enzymatic hydrolysis products will assemble in situ with the target delivery substance and bile salts in the intestine, continuing to complete drug transport and absorption until they are completely metabolized into steviol.

[0472] (3) Evaluation of the transintestinal mucosal permeability of celecoxib and MRTX1133 supramolecular composites

[0473] MRTX1133 is specifically targeted against KRAS G12D A new drug developed for cancers caused by KRASG mutations is not yet on the market, but according to the Discovery of Prodrug of MRTX1133 as an Oral Therapy for Cancers with KRASG 12DMutation (https: / / doi.org / 10.1021 / acsomega.3c00329) conducted preclinical studies on MRTX1133. The permeability coefficient Papp results of the Caco-2 cell line are as follows:

[0474] A→B:<0.48×10 -6 cm / s, and B→A:8.05×10 -6 cm / s,efflux ratio:>16.8;

[0475] The absolute oral bioavailability in mice is only 0.5%. P-gp efflux is the main problem that causes the oral inability to absorb MRTX1133.

[0476] The experiment was conducted using the everted intestinal sac method, following the same experimental procedures as previously described. To evaluate the ability of the supramolecular composition to enter the intestine after oral administration, the metabolites in the supramolecular composition, after being metabolized by β-glucosidase, undergo in situ assembly, continuing to transport and absorb the target delivery substance. Therefore, different intestinal segments were used for the experiments based on different formulations. The original phosphate buffer was replaced with simulated intestinal fluid before a meal. A solution of intestinal flora isolated and cultured from human feces was added to the simulated intestinal fluid. Prior to the experiment, the simulated intestinal fluid with the added flora solution was incubated at 37°C for 24 hours.

[0477] SD rats were fasted for 12 hours before the experiment and had free access to water. After being sacrificed by cervical dislocation, the abdominal cavity was opened, and the jejunum, ileum, and colon were quickly removed. After being rinsed with cold Krebs-Ringer solution, the rats were rotated with a glass rod and ligated with surgical suture to form intestinal sacs. The intestinal sacs were fixed, and 2 mL of Krebs solution was injected into each sac using a syringe. The intestinal segments were then placed in a beaker containing 250 mL of pre-prandial intestinal fluid containing a fixed concentration of celecoxib or MRTX1133, or pre-prandial intestinal fluid containing a specific intestinal flora (MF) at 37°C. The supramolecular compositions were converted based on the measured content. Both celecoxib and MRTX1133 supramolecular compositions were prepared using the supramolecular compositions of formulas F48, F49, and F52, and the initial concentration was 50 μg / mL. Stirring was initiated, and a CO₂-O₂ (5:95) gas mixture was aerated into the medium during incubation. The intestinal sacs were then placed into the incubation compartments according to their respective groups. At 30, 60, 90, and 120 minutes, 200 μL of the fluid from each intestinal sac was aspirated as the test solution, and the same volume of simulated intestinal fluid incubated at 37°C or simulated intestinal fluid containing intestinal flora was added. At the end of the experiment, the intestinal sac area was measured. Protein was precipitated with three volumes of acetonitrile, followed by centrifugation at 13,000 rpm and 37°C for 10 minutes. 1 mL of the supernatant was nitrogen-purged to a volume of 0.5 mL and analyzed by high-performance liquid chromatography for celecoxib or MRTX1133 concentration. The content was calculated using the external standard method. At the end of the experiment, the length and width of the dissected intestinal segments were measured, and the permeation area was calculated.

[0478] According to the permeation drug concentration measured at each sampling point, the cumulative permeation amount of drug Q (ng), drug permeation rate constant K and apparent permeability coefficient (P app， cm·s -1 ), the results are shown in Table 38 below:

[0479] Table 38. Apparent permeability coefficient investigation by everted intestinal sac method (n=3)

[0480] In summary, positively charged supramolecular compositions have better permeability in the small intestine, while negatively charged supramolecular compositions have better permeability in the colon. In the presence of intestinal bacterial metabolic enzymes, supramolecular compositions containing the STVB-Arga carrier are hydrolyzed by amidase to STVB, which becomes negatively charged. Therefore, the apparent permeability coefficients of formulations F-48 and F-52 in the colon did not decrease significantly. Compared with the API, the apparent permeability coefficient of the celecoxib supramolecular composition is over 100 times that of the API; while the apparent permeability coefficient of the MRTX1133 supramolecular composition is 136-196 times that of the API.

[0481] The results of the everted intestinal sac method show that if β-glucosidase hydrolyzes some of the sugar residues of the carrier without in situ assembly, the apparent permeability coefficient will decrease significantly. However, the experimental results are just the opposite, indicating that when RBDS-D, RBDS-B, or STVB are obtained after partial enzymatic hydrolysis, they will reassemble in situ. Therefore, the apparent permeability coefficient does not show a downward trend.

[0482] Example 13: Supramolecular composition of macitentan and finerenone

[0483] According to the FDA Orange Book, macitentan is marketed as a film-coated tablet (trade name: OPSUMIT). In vitro dissolution testing using 900 ml of pH 6.8 phosphate buffer (containing 0.1% cetyltrimethylammonium bromide (CTAB)) as the dissolution medium, using a paddle method at 75 rpm, achieved 90% dissolution in 45 minutes. However, the peak dissolution time of approximately 8 hours indicates a very low in vivo dissolution rate. Macitentan is a substrate for the cytochrome P450 CYP3A4 isoenzyme. The chemical structure of its primary active metabolite, ACT-132577 (5-(4-bromophenyl)-4-[2-(5-bromopyrimidin-2-yl)oxyethoxy]-6-(sulfamethoxypyrimidine), is as follows:

[0484] From a chemical structure perspective, while ACT-132577 (LogP: ~2.6) has a lower LogP than macitentan (LogP: ~3.7) and is slightly less lipophilic, both are poorly soluble drugs. The following analysis uses macitentan as a model drug, and the same applies to ACT-132577.

[0485] (1) The formula of the supramolecular composition is as shown in Table 39:

[0486] Take an appropriate amount of macitentan and add acetone-dichloromethane (4:1, V / V) to dissolve it into a solution containing approximately 250 mg of macitentan per 1 ml, and set aside.

[0487] Dissolve an appropriate amount of finerenone in methanol-dichloromethane (3:1, v / v) to a solution containing approximately 150 mg of finerenone per 1 ml. Set aside. Weigh the carrier, polymer, and surfactant, add 10 ml of pH 6.8 phosphate buffer, and sonicate to dissolve. While stirring, slowly add the organic phase dropwise to the aqueous phase until noticeable turbidity appears. Stop the addition and continue stirring until the organic solvent smell disappears. Centrifuge at 13,000 rpm at room temperature for 5 minutes after approximately 30-60 minutes to separate the supernatant.

[0488] Macitentan was prepared according to formula F53-F57, and finerenone was prepared according to formula F58-F61.

[0489] (2) Evaluation of supramolecular composite properties

[0490] The above solutions were placed in a sealed container at room temperature for 72 hours to observe whether phase separation occurred. Supramolecular composition solutions that did not undergo phase separation were sampled for content testing and subjected to a dilution tolerance test. The dilution media were pre-meal simulated gastric fluid and pre-meal simulated intestinal fluid, respectively. An appropriate amount of the diluted solution was taken. The dilution test was performed in a dissolution cup. 1ml of the supramolecular composition solution was accurately measured and first added to 150ml of 37℃ simulated gastric fluid. After 30 minutes, 2ml was taken and the remainder was adjusted to simulated intestinal fluid with a regulating solution. Stirring was continued for 2 hours before sampling and testing.

[0491] Sample Preparation: 2 mL of sample solution was centrifuged at 37°C, 13,000 rpm, for 5 min. 0.2 mL of the supernatant was diluted to 5 mL with 70% acetonitrile-water. Simultaneously, 2 mL of the same medium was added to the dissolution vessel. The target substance content in the supernatant was determined by HPLC and calculated based on the theoretical concentration to determine dilution tolerance. The results are shown in Table 40 below.

[0492] Table 40. Dilution tolerance experiment of supramolecular composition of macitentan and finerenone

[0493] In summary, the supramolecular compositions prepared according to the above formulations and processes were tested for dilution tolerance in simulated gastrointestinal fluid before a meal. The macitentan-F55 supramolecular composition showed poor dilution tolerance, with dilution in simulated gastric fluid causing the supramolecular composition to be destroyed. Even after reconstitution in simulated intestinal fluid, the composition was unable to reassemble. However, both macitentan-F56 and -F57 were able to withstand dilution in simulated intestinal fluid before a meal. Finerenone-F58 showed a small amount of sediment at the bottom of the centrifuge tube after dilution in simulated gastric fluid before a meal, and the assay result was slightly low. However, after consolidation in simulated intestinal fluid, the assay result was 98.54% of the theoretical value. Finerenone-F61 also showed sediment at the bottom of the tube after dilution in simulated gastric fluid before a meal, and the assay result was only 67.44% of the theoretical value, indicating that some of the supramolecular composition was destroyed. However, after consolidation in simulated intestinal fluid, the assay result was 97.20% of the theoretical value, indicating that the previously disintegrated nanoparticles reassembled in the intestinal fluid.

[0494] Example 14 Nutrient supramolecular composition

[0495] Supramolecular compositions were prepared using vitamin K1 as the target delivery agent.

[0496] (1) Preparation of vitamin K1 supramolecular self-assembly complex according to the formula composition of Table 41

[0497] (2) Preparation process

[0498] Organic phase: Dissolve vitamin K1 in ethanol to prepare a solution containing approximately 30 mg per 1 ml.

[0499] Aqueous Phase: Dissolve the carrier or polymer in 10 ml of pH 6.8 phosphate buffer. Once dissolved and clarified, slowly add the organic phase dropwise to the aqueous phase while stirring until noticeable turbidity appears. Stop the addition and continue stirring for 30-60 minutes. Remove the organic solvent using a rotary evaporator at 45°C, and centrifuge the aqueous phase at 15,000 rpm for 5 minutes. Analyze the vitamin K1 concentration by HPLC. Freeze-dry for 48 hours to obtain a vitamin K1 supramolecular composition powder.

[0500] The supramolecular compositions of formulas F-65, F-66, F-67, and F-70 were redissolved in water and the conditions after redissolution were examined. The results are shown in Table 42 below.

[0501] Table 42 Vitamin K1 supramolecular composition re-dissolution

[0502] In summary, the synergistic effect of the combination of carrier and polymer is significantly better than that of the single carrier or the single polymer; the synergistic effect of the combination of two carriers and then the polymer is significantly better than that of a single carrier and polymer, and has a higher drug loading capacity.

Claims

1. A kit for forming a supramolecular self-assembly complex, comprising: package 1, package 2 and package 3, wherein Package 1 comprises one or more carriers I, wherein the carrier I provides positive charge or negative charge under physiological conditions, and the carrier I is as shown in structural formula (1-1) or (1-2), Wherein R1 in the structural formula (1-1) / (1-2) is selected from the following groups: wherein R3 is a straight and / or branched alkane having 1 to 10 carbon atoms (preferably 1-2 carbon atoms), CPPs are cell-penetrating peptides, (5) amino acids or short peptides consisting of 2-20 amino acids. PDA is polydopamine. Wherein R4 is OH and / or NH2, Wherein R4 is OH group, and / or NH2, Wherein R4 is OH group, and / or NH2, Wherein R4 is OH group, and / or NH2, or Where n=0-9, X=OH and / or NH2, Wherein R2 in the structural formula (1-1) is selected from the following groups: Wherein R2 in the structural formula (1-2) is selected from the following groups: Package 2 comprises one or more polymers, the polymers having groups that can dissociate under physiological conditions, and / or having more than 10 groups that can provide hydrogen donors and / or hydrogen acceptors, and Package 3 comprises one or more carriers II, wherein the carrier II is composed of a covalently linked unit A and a unit B, wherein unit A is a hydrophilic portion, and unit B is a hydrophobic portion, wherein unit A is located on one side or both sides of unit B, and when unit A is located on both sides of unit B, the units A may be the same or different; wherein the number ratio of unit A to unit B is (0:1)-(7:1), Wherein unit B is selected from one or more of B1, or one or more of B2, wherein: (1) B1, molecular formula is (C5H8) n The compound represented by, and its alcohol, aldehyde, ketone, carboxylic acid or ester derivatives, wherein n=4 or 6; (2) B2, a compound having two benzene rings (A-ring and B-ring) connected to each other by a central three-carbon chain, with 2-phenylchromone as the basic parent nucleus and a basic skeleton of C6-C3-C6, and its derivatives (such as halides or amino acid esters, the halides are such as chlorine or fluorine, and the amino acid esters are such as esters formed by the hydroxyl group on C6 and amino acids); The unit A is selected from one or more of A1 or a derivative of A1, wherein: A1 is selected from D-type or L-type monosaccharide, disaccharide, trisaccharide or a combination thereof, The monosaccharide is selected from the group consisting of glucose, rhamnose, galactose, arabinose, xylose, mannose, fucose, apiose, glucuronic acid, glucosamine, galacturonic acid, acetylamino sugar and combinations thereof, and the relative configuration of each glycosyl terminal carbon can be α-type or β-type; The disaccharide is selected from the group consisting of sophorose (glc1-2glc), gentiobiose (glC1-6glc), rutinose (rha1-6glc), neohesperidose (rha1-6glC), acaciabiose (rha1-6gal) and combinations thereof; A trisaccharide selected from the group consisting of gentiotriose, sophorotriose (glc1-2glc1-2glc) and a combination thereof; The derivative of A1 is an acylated product of A1, such as 2-acetylglucose and caffeoylglucose.

2. The kit according to claim 1, wherein the amino acids in the carrier I are selected from alanine, arginine, aspartic acid, cysteine, glutamic acid, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, serine, threonine, tryptophan, tyrosine, valine, or their corresponding amino amides (e.g., arginine amide, histidine amide, homoarginine), and polyamino acids composed thereof, preferably L-configuration amino acids, Alternatively, the number of amino acids in the cell-penetrating peptide in the vector I is less than 20.

3. The kit according to claim 1, wherein the carrier I is as shown in structural formula (1-1).

4. The kit according to claim 1, wherein the ester derivative of B1 is an ester formed from a carboxylic acid derivative of B1, such as gluconate ester, polyethylene glycol ester, propylene glycol ester, methyl ester, ethyl ester or 1-aminoglycine ester.

5. The kit according to claim 1, characterized in that The polymer in the package 2 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, 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 combinations thereof.

6. The kit according to any one of claims 1 to 5, characterized in that The hydrophobic unit B is a tetracyclic diterpene, a tetracyclic triterpene, a pentacyclic triterpene, or an alcohol, aldehyde, ketone, carboxylic acid or ester derivative thereof.

7. The kit according to any one of claims 1 to 6, characterized in that The hydrophobic unit B is kaurene-type tetracyclic diterpenes and their derivatives (such as alcohols, aldehydes, ketones, carboxylic acids or esters).

8. The kit according to any one of claims 1 to 7, characterized in that The hydrophobic unit B is one or more of dammarane type, gansuline type, cycloartane type, lanolin type, cucurbitane type, azadirachtin type and prototerpane type, and their derivatives (such as alcohols and esters), more preferably cucurbitane type and its derivatives (such as alcohols and esters).

9. The kit according to any one of claims 1 to 8, characterized in that The hydrophobic unit B is one or more of oleanane type, ursane type, lupeane type, suberane type, Hobane type and iso-Hobane type, and derivatives thereof (such as alcohols, ketones, carboxylic acids and esters).

10. The kit according to any one of claims 1 to 9, characterized in that (1) When unit B1 is a kaurene-type tetracyclic diterpene or a pharmaceutically acceptable salt thereof (such as a sodium salt, a potassium salt, an ammonium salt), a hydrate of a kaurene-type tetracyclic diterpene or a pharmaceutically acceptable salt thereof (such as a hydrate formed with 0.5, 1, 1.5, 2, 2.5 or 3 crystal waters), the carrier II has the structure of formula (2); Wherein R1 and R2 represent unit A, and R1 and R2 exist simultaneously, and R1, R2 are independently selected from the following groups: or (2) When unit B1 is a cucurbitane-type tetracyclic triterpene or a pharmaceutically acceptable hydrate or solvate thereof, the carrier II has the structural formula (3): Wherein R1 and R2 represent unit A, and R1 and R2 exist simultaneously or alone, and R1 is selected from the following groups: R2 is selected from the following groups: R3 is selected from R4 is selected from 11. The kit according to any one of claims 1 to 10, characterized in that The unit B is B2, and is selected from the group consisting of flavonols (as represented by structural formula (4-1)), or dihydrochalcone (as represented by structural formula (4-2)), and a combination thereof, wherein the structure of structural formula (4-1) or structural formula (4-2) is as follows: The structure of structural formula (4-1) is as follows: Wherein R1 is selected from R2 is selected from H, R3 is selected from H, or when R2 is non-hydrogen substituted, R3 is absent, and the dotted line in the structural formula represents a double bond R4 is selected from H or R5 is selected from H or The structure of structural formula (4-2) is as follows: Wherein R1 is selected from R2 is selected from R3 is selected from H or R4 is selected from H or 12. The kit according to any one of claims 1 to 11, characterized in that The carrier I is selected from the group consisting of: (1) (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), (2) (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), (3) (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), (4) (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-methylenetetradecahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxamide (STVB-APL), (5) (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), (6) (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-carboxylate sodium (STVB-Na), (7) Any combination of the compounds shown in (1) to (6).

13. The kit according to any one of claims 1 to 12, characterized in that The carrier II is selected from the group consisting of the following compounds: (1) [(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](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 (RBDS-M), (2) (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](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 (RBDS-D), (3) [(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl](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 (RBDS-A), (4) (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-carboxylic acid (RBDS-B), (5) [(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl] (1R,4S,5R,9S,10R,13S)-13-[(2S,3R,4S,5R,6R)-5-hydroxy-6-(hydroxymethyl)-4-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl] 4R,5R,6S)-3,4,5-trihydroxy-6-methyloxa-2-yl]oxyoxa-2-yl]oxy-5,9-dimethyl-14-methylenetetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylate (RBDS-C), (6) [(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl](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-carboxylate (STVS), (7) [(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl](1R,4S,5R,9S,10R,13S)-13-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4R),5R,6S)-3,4,5-trihydroxy-6-methyloxa-2-yl]oxyoxa-2-yl]oxy-5,9-dimethyl-14-methylenetetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylate (DCS-A), (8) (4R,4aS,6aR,9S,11aR,11bS)-9-(((2S,3R,4S,5R,6R)-5-hydroxy-6-(hydroxymethyl)-4-(((2S,3R,4S),5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-3-((((2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyltetrahydro-2H-pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxylic acid (RBDS-C-Acid), (9) (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), (10) [(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl](1R,4S,5R,9S,10R,13S)-5,9-dimethyl-14-methylene-13-[(2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxa-2-yl]oxytetracyclo[11.2.1.01,10.04,9]hexadecane-5-carboxylate (RBSD), (11)(2R,3S,4S,5R,6R)-2-(hydroxymethyl)-6-({[(2R,3S,4S,5R,6S)-3,4-dihydroxy-6-{[(3R,6R)-2-hydroxy-6-[(1R,3aS,3bS,7S,9aR,9bR,10R,11aR)-10-hydroxy-7-{[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-({[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)-3,4,5,6-tetrahydro-2H-pyran-2-yl]oxy}methyl)-3,4,5, 6-tetrahydro-2H-pyran-2-yl]oxy}-3a,6,6,9b,11a-pentamethyl-2,3,3a,3b,4,6,7,8,9,9a,9b,10,11,11a-tetradecahydro-1H-cyclopenta[1,2-a]phenanthrene-1-yl]-2-methylhept-3-yl]oxy}-5-{[(2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)-3,4,5,6-tetrahydro-2H-pyran-2-yl]oxy}-3,4,5,6-tetrahydro-2H-pyran-2-yl]methyl}oxy)tetrahydropyran-3,4,5-triol (MGSD-V), (12)(3S,8S,9R,10R,13R,14S,17R)-17-((2R,5R)-5-((((2S,3R,4S,5S,6R)-4,5-dihydroxy-3-(((2S,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)-6- ((((2R,3R,4S),5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)-6-hydroxy-6-methylheptane-2-yl)-4,4,9,13,14-pentamethyl-3-(((2R,3R,4S,5S,6R)-3,4,5-trihydroxy- 6-((((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)-1,2,3,4,7,8,9,10,12,13,14,15,16,17-tetradecahydro-11H-cyclopenta[a]phenanthrene-11-one (OMGSD-V), (13)(2S,3R,4S,5S,6R)-2-(((2S,3R,4S,5S,6R)-4,5-dihydroxy-2-(((3R,6R)-2-hydroxy-6-((3S,8S,9R,10R,11R,13R,14S,17R)-11-hydroxy-4,4,9,13,14-pentamethyl-3-(((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)- 2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetrahydro-1H-cyclopenta[a]phenanthren-17-yl)-2-methylheptane-3-yl)oxy)-6-((((2R,3R,4S,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)tetrahydro-2H-pyran-2-yl)oxy)methyl)tetrahydro-2H-pyran-3-yl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (SMSD-I), (14) (2R,3R,4S,5S,6R)-2-(((3S,8R,9R,10S,11R,13R,14S,17R)-17-((2R,5R)-5,6-dihydroxy-6-methylheptane-2-yl)-11-hydroxy-4,4,13,14-tetramethyl-2,3,4,7,8,9,10,11,12,13,14,15,16,17-tetradecahydro-1H-cyclopenta[a]phenanthrene-3-yl)oxy)-6-(hydroxymethyl)tetrahydro-2H-pyran-3,4,5-triol (MGSD-IE1), (15) (2S)-7-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-2-yl]oxy-5-hydroxy-2-(3-hydroxy-4-methoxyphenyl)-2,3-dihydrochromen-4-one (NHPD), (16) 1-[4-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxy-2-yl]oxyoxan-2-yl]oxy-2,6-dihydroxyphenyl]-3-(3-hydroxy-4-methoxyphenyl)propan-1-one (NHDC), (17) 1-[4-[(2S,3R,4S,5S,6R)-4,5-dihydroxy-6-(hydroxymethyl)-3-[(2S,3R,4R,5R,6S)-3,4,5-trihydroxy-6-methyloxan-2-yl]oxyoxan-2-yl]oxy-2,6-dihydroxyphenyl]-3-(4-hydroxyphenyl)propan-1-one (NRGDC), (18) methyl (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)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxylate (RBDS-B-Methyl), (19) Ethyl (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-pyrrolidone) pyran-2-yl)oxy)tetrahydro-2H-pyran-2-yl)oxy)-4,11b-dimethyl-8-methylenetetrahydro-6a,9-methylcyclohepta[a]naphthalene-4-carboxylate (RBDS-B-Ethyl), (20) conjugates of steviol glycosides (stevioside, RBDS-B, STVB, RBSD)-HA, and (21) Any combination of the compounds shown in (1) to (20).

14. The kit according to any one of claims 1 to 13, wherein The kit further comprises a package 4, wherein the package 4 comprises an active ingredient, wherein the active ingredient is a compound, preferably a drug, a monomer component or component extract of a natural product, a nutrient, a contrast agent and / or a diagnostic agent.

15. The kit according to claim 14, wherein The mass ratio of the active ingredient to the sum of the carrier I and the carrier II is 1:0.5-1:50 (preferably 1:1, 1:5, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40 or 1:45), the carrier I accounts for 0%-75% (preferably 0%-70%, 0%-65%, 0%-60%, 0%-55%, 0%-50%, 0-40%, 0-30%, 0-20% or 0-10%) of the total mass of the carrier I and the carrier II; the mass ratio of the active ingredient to the polymer is 1:0-1:25 (preferably 1:0-1:20, 1:0-1:15, 1:0-1:10, 1:0-1:5).

16. The kit of claim 14, wherein the drug is selected from small molecule compound drugs, The nutrient is a vitamin or coenzyme Q10, preferably a vitamin selected from the group consisting of one or more of vitamin A, B vitamins, vitamin E, vitamin K, and vitamin D; The contrast agent is selected from tumor fluorescent agents, CT tracers (such as iodine nanocontrast agents, gold nanocontrast agents, tantalum oxide nanocontrast agents, bismuth nanocontrast agents or lanthanide nanocontrast agents), MRI tracers (such as paramagnetic contrast agents, ferromagnetic contrast agents or supermagnetic contrast agents) and radionuclide tracers.

17. The kit according to any one of claims 14 to 16, wherein the active ingredient is a free acid, a free base, a salt (e.g., hydrochloride, sulfate, phosphate, fumarate, tartrate, hemifumarate, maleate, aspartate, palmitate, tartrate, bitartrate, methanesulfonate, citrate, toluenesulfonate, bromate, sulfite, carbonate, bicarbonate, malate, succinate, ethanesulfonate, acetate, formate, sodium salt, iron salt, potassium salt, The invention relates to an active ingredient in the form of a hydrate, a solvate, a calcium salt, an ammonium salt or a magnesium salt, or a co-crystal (the co-crystal is a co-crystal prepared from the active ingredient and a co-crystal ligand, and the co-crystal ligand used is a natural amino acid, caffeine, saccharin, fumaric acid, tartaric acid, oxalic acid, malonic acid, maleic acid, aspartic acid, tartaric acid, bitartrate, pyroglutamic acid, nicotinic acid, nicotinamide, an organic base such as meglumine, tromethamine, isopropanolamine), or a hydrate, a solvate, or a solvate hydrate of the free acid, the free base, the salt or the co-crystal.

18. The kit according to any one of claims 14 to 17, characterized in that The package 4 also includes: (1) pH adjuster, (2) Surfactants and / or co-surfactants, such as gum arabic, gum tragacanth, gelatin, propylene glycol monostearate, glyceryl monostearate, ethylene glycol distearate, diglyceryl monooleate, sodium lauryl sulfate, sorbitan fatty acid (Span 20, Span 40, Span 60, Span 65, Span 80, Span 83 or Span 85), potassium oleate, sodium oleate, triethanolamine oleate, lecithin, sucrose esters, polyoxyethylene-polyoxypropylene copolymer (poloxamer), propylene laurate, polysorbate (Tween 20, Tween 40, Tween 60, Tween 65, Tween 80 or Tween 85), polyoxyethylene monostearate, polyoxyethylene stearate, polyoxyethylene 400 monolaurate, polyoxyethylene 400 monostearate, polyoxyethylene 400 monooleate, polyoxyethylene lauryl alcohol ether, polyoxyethylene cetyl alcohol ether, vitamin E succinate polyethylene glycol ester (TPGS), betaine, polyoxyethylene castor oil, polyoxyethylene hydrogenated castor oil, glycerin, sorbitol, polyethylene glycol, one or more; and / or (3) Bile salts, such as sodium glycocholate, sodium deoxyglycocholate, sodium chenodeoxyglycocholate, sodium ursodeoxyglycocholate, sodium taurocholate, sodium taurodeoxycholate, sodium taurochenodeoxycholate, and sodium deoxycholate.

19. A supramolecular self-assembled complex comprising the carrier I, carrier II and polymer as defined in any one of claims 1 to 13, and the active ingredient as defined in any one of claims 14 to 17.

20. A pharmaceutical composition comprising the supramolecular self-assembled complex according to claim 19, and a pharmaceutical excipient, wherein the pharmaceutical excipient is selected from the group consisting of a filler, a disintegrant, a binder, a lubricant, a glidant, an emulsifier, a flavoring agent or a taste-masking agent, a surfactant, a co-surfactant, a preservative, a propellant, a pH regulator, an osmotic pressure regulator, a gel / ointment base, a suspending agent, a coating component, a pressure-sensitive adhesive, a penetration enhancer, and a combination thereof.

21. The pharmaceutical composition according to claim 20, which is in the form of tablets, capsules, suspensions, solutions, patches, creams, gels, emulsions, eye drops, injections, oral capsules, suppositories, implants, powders, granules or implant products.

22. A method for preparing the supramolecular self-assembly complex according to claim 19, comprising dissolving and mixing the carrier I, carrier II and polymer defined in any one of claims 1 to 14, and the active ingredient defined in any one of claims 14 to 17, respectively, wherein the carrier I and carrier II are added and mixed simultaneously, or are mixed separately with the polymer and / or the active ingredient and then mixed together.

23. A composition for delivering an active ingredient, comprising the supramolecular self-assembled complex defined in item 19, and a pharmaceutically or food-acceptable carrier, excipient, diluent, adjuvant, vehicle or a combination thereof.

24. Use of the kit according to any one of claims 1 to 18 or the supramolecular self-assembled complex according to claim 19 in delivering active ingredients, preferably in delivering to mammals (such as humans, rats, mice, primates).

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