CN116144031A - Cyclodextrin derivatives, and preparation method and application thereof - Google Patents
Cyclodextrin derivatives, and preparation method and application thereof Download PDFInfo
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- CN116144031A CN116144031A CN202310019403.XA CN202310019403A CN116144031A CN 116144031 A CN116144031 A CN 116144031A CN 202310019403 A CN202310019403 A CN 202310019403A CN 116144031 A CN116144031 A CN 116144031A
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- mixed solution
- cyclodextrin
- vitamin
- peg
- polyethylene glycol
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- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical class O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 title claims abstract description 119
- 238000002360 preparation method Methods 0.000 title claims abstract description 18
- 229920000858 Cyclodextrin Polymers 0.000 claims abstract description 56
- 150000001875 compounds Chemical class 0.000 claims abstract description 51
- 229940045997 vitamin a Drugs 0.000 claims abstract description 43
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 claims abstract description 38
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 claims abstract description 38
- 235000019155 vitamin A Nutrition 0.000 claims abstract description 38
- 239000011719 vitamin A Substances 0.000 claims abstract description 38
- -1 vitamin A modified cyclodextrin Chemical class 0.000 claims abstract description 34
- 239000003814 drug Substances 0.000 claims abstract description 15
- 239000011259 mixed solution Substances 0.000 claims description 106
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims description 78
- 229920001223 polyethylene glycol Polymers 0.000 claims description 66
- YMWUJEATGCHHMB-UHFFFAOYSA-N Dichloromethane Chemical compound ClCCl YMWUJEATGCHHMB-UHFFFAOYSA-N 0.000 claims description 45
- 239000002202 Polyethylene glycol Substances 0.000 claims description 38
- HARGZZNYNSYSGJ-UHFFFAOYSA-N 1,2 dihydrotanshinquinone Natural products C1=CC2=C(C)C=CC=C2C(C(=O)C2=O)=C1C1=C2C(C)CO1 HARGZZNYNSYSGJ-UHFFFAOYSA-N 0.000 claims description 37
- HARGZZNYNSYSGJ-JTQLQIEISA-N Dihydrotanshinone I Chemical compound C1=CC2=C(C)C=CC=C2C(C(=O)C2=O)=C1C1=C2[C@@H](C)CO1 HARGZZNYNSYSGJ-JTQLQIEISA-N 0.000 claims description 37
- 238000000502 dialysis Methods 0.000 claims description 30
- 238000000034 method Methods 0.000 claims description 30
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-Dimethylaminopyridine Chemical compound CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 claims description 22
- LMDZBCPBFSXMTL-UHFFFAOYSA-N 1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide Substances CCN=C=NCCCN(C)C LMDZBCPBFSXMTL-UHFFFAOYSA-N 0.000 claims description 21
- 238000003756 stirring Methods 0.000 claims description 21
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 claims description 20
- FPQQSJJWHUJYPU-UHFFFAOYSA-N 3-(dimethylamino)propyliminomethylidene-ethylazanium;chloride Chemical compound Cl.CCN=C=NCCCN(C)C FPQQSJJWHUJYPU-UHFFFAOYSA-N 0.000 claims description 18
- 239000001116 FEMA 4028 Substances 0.000 claims description 18
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- 238000006243 chemical reaction Methods 0.000 claims description 18
- 235000011175 beta-cyclodextrine Nutrition 0.000 claims description 15
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- NQTADLQHYWFPDB-UHFFFAOYSA-N N-Hydroxysuccinimide Chemical compound ON1C(=O)CCC1=O NQTADLQHYWFPDB-UHFFFAOYSA-N 0.000 claims description 12
- 238000004108 freeze drying Methods 0.000 claims description 12
- 239000000243 solution Substances 0.000 claims description 12
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- FALRKNHUBBKYCC-UHFFFAOYSA-N 2-(chloromethyl)pyridine-3-carbonitrile Chemical compound ClCC1=NC=CC=C1C#N FALRKNHUBBKYCC-UHFFFAOYSA-N 0.000 claims description 6
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 6
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 claims description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 6
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- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- 239000007864 aqueous solution Substances 0.000 claims description 3
- MTHSVFCYNBDYFN-UHFFFAOYSA-N diethylene glycol Chemical compound OCCOCCO MTHSVFCYNBDYFN-UHFFFAOYSA-N 0.000 claims description 3
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- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 18
- KXNYCALHDXGJSF-UHFFFAOYSA-N dihydroisotanshinone I Natural products CC1=CC=CC2=C(C(C=3OCC(C=3C3=O)C)=O)C3=CC=C21 KXNYCALHDXGJSF-UHFFFAOYSA-N 0.000 description 11
- VOFUROIFQGPCGE-UHFFFAOYSA-N nile red Chemical compound C1=CC=C2C3=NC4=CC=C(N(CC)CC)C=C4OC3=CC(=O)C2=C1 VOFUROIFQGPCGE-UHFFFAOYSA-N 0.000 description 10
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- 125000002496 methyl group Chemical group [H]C([H])([H])* 0.000 description 3
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- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 2
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- 102000000340 Glucosyltransferases Human genes 0.000 description 1
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- RNFNDJAIBTYOQL-UHFFFAOYSA-N chloral hydrate Chemical compound OC(O)C(Cl)(Cl)Cl RNFNDJAIBTYOQL-UHFFFAOYSA-N 0.000 description 1
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Images
Classifications
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- C—CHEMISTRY; METALLURGY
- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08G—MACROMOLECULAR COMPOUNDS OBTAINED OTHERWISE THAN BY REACTIONS ONLY INVOLVING UNSATURATED CARBON-TO-CARBON BONDS
- C08G81/00—Macromolecular compounds obtained by interreacting polymers in the absence of monomers, e.g. block polymers
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L29/00—Foods or foodstuffs containing additives; Preparation or treatment thereof
- A23L29/30—Foods or foodstuffs containing additives; Preparation or treatment thereof containing carbohydrate syrups; containing sugars; containing sugar alcohols, e.g. xylitol; containing starch hydrolysates, e.g. dextrin
- A23L29/35—Degradation products of starch, e.g. hydrolysates, dextrins; Enzymatically modified starches
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
- A61K47/6951—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes using cyclodextrin
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2250/00—Food ingredients
- A23V2250/50—Polysaccharides, gums
- A23V2250/51—Polysaccharide
- A23V2250/5112—Cyclodextrin
Abstract
The invention discloses cyclodextrin derivatives, a preparation method and application thereof, and belongs to the technical field of pharmacy. The PEG modified cyclodextrin can improve the solubility of the cyclodextrin, improve the in vivo drug absorption quantity and obviously improve the drug distribution quantity of the liver part; the vitamin A modified cyclodextrin can facilitate the clathrate compound to be taken up by hepatic stellate cells of liver, and can improve the liver targeting of the medicine, thereby improving the curative effect and reducing the toxic and side effects.
Description
Technical Field
The invention relates to the technical field of pharmacy, in particular to cyclodextrin derivatives, and a preparation method and application thereof.
Background
Cyclodextrin (CD) is a generic term for a series of cyclic oligosaccharides produced by amylose under the action of Cyclodextrin glucosyltransferase produced by Bacillus. Cyclodextrins, especially amino beta-cyclodextrin, have the characteristic of being hydrophobic on the inside and hydrophilic on the outside. The drug molecules with the size matched with the hydrophobic cavity partially or completely enter the cyclodextrin cavity through non-covalent bond action, so that the cyclodextrin inclusion compound is formed. Cyclodextrin is used as a host molecule, and can self-assemble with small molecular drugs used as guest molecules to form an inclusion compound through interaction of the host and the guest, and the hydrophobic property of the 6-position substituent chain of the cyclodextrin can further improve the capability of forming the inclusion compound between a hydrophobic cavity and the guest drug molecules.
Liver fibrosis is a pathological process of abnormal hyperplasia of connective tissue of liver, and clinically common causes include hepatitis virus infection, cholestasis, alcoholism, nonalcoholic steatohepatitis and the like. Any liver injury has liver fibrosis process in the liver repairing and healing process, and if injury factors cannot be removed for a long time, the fibrosis process can develop into liver cirrhosis and even liver cancer after long-term persistence. Liver fibrosis is thus an important link affecting the prognosis of liver disease, and early intervention treatment can reverse liver fibrosis. However, there are few currently clinically available anti-fibrotic therapeutic strategies and there is an urgent need for effective therapeutic agents. The existing cyclodextrin does not have liver targeting, so that an inclusion compound formed by the cyclodextrin serving as a carrier and an anti-liver fibrosis drug is difficult to specifically target liver tissues, and the high-efficiency anti-liver fibrosis capacity is exerted.
Disclosure of Invention
Aiming at the technical problems in the prior art, the invention provides the cyclodextrin derivative with the liver targeting characteristic, the preparation method and the application thereof, and the concentration of an inclusion compound formed by the cyclodextrin derivative and a small molecular medicine in the liver is improved, so that the anti-hepatic fibrosis effect of the inclusion compound is improved.
The invention discloses a cyclodextrin derivative, which is PEG modified cyclodextrin or vitamin A modified cyclodextrin, wherein the PEG modified cyclodextrin has the structural formula:
the vitamin A modified cyclodextrin has the structural formula:
preferably, the preparation method of the PEG modified cyclodextrin comprises the following steps:
step S1: obtaining dicarboxylated polyethylene glycol;
step S2: dissolving dicarboxylated polyethylene glycol, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide in dimethyl sulfoxide to obtain a first mixed solution;
step S3: dissolving amino beta cyclodextrin in dimethyl sulfoxide to obtain a second mixed solution;
step S4: dropwise adding the second mixed solution into the first mixed solution, and stirring for reaction to obtain a third mixed solution;
step S5: and purifying the third mixed solution by a dialysis method to obtain the PEG modified cyclodextrin.
Preferably, the preparation method of the PEG modified cyclodextrin comprises the following steps:
step S2: 880mg of dicarboxylated polyethylene glycol, 153.4mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and 92mg of N-hydroxysuccinimide were dissolved in 20mL of dimethyl sulfoxide to obtain a first mixed solution;
step S3: 453.6mg of amino beta cyclodextrin is dissolved in 10mL of dimethyl sulfoxide to obtain a second mixed solution, wherein the amino beta cyclodextrin is mono-6-O-amino-beta cyclodextrin;
step S4: dropwise adding the second mixed solution into the first mixed solution at 25 ℃, and stirring and reacting for 72 hours to obtain a third mixed solution;
step S5: and (3) dialyzing the third mixed solution for 48 hours through a 3500DA dialysis bag, and freeze-drying the liquid in the dialysis bag to obtain the PEG modified cyclodextrin.
Preferably, the vitamin A modified cyclodextrin is prepared by the following steps:
step 101: obtaining the synthesis of the dicarboxylated polyethylene glycol;
step 102: dissolving dicarboxylated polyethylene glycol, 4-dimethylaminopyridine and 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride in dimethyl sulfoxide to obtain a fourth mixed solution;
step 103: dissolving vitamin A in dimethyl sulfoxide to obtain a fifth mixed solution;
step 104: dropwise adding the fifth mixed solution into the fourth mixed solution, and stirring for reaction to obtain a sixth mixed solution;
step 105: purifying the sixth mixed solution by a dialysis method to obtain vitamin A modified polyethylene glycol;
step 106: dissolving vitamin A modified polyethylene glycol, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide in dimethyl sulfoxide to obtain a seventh mixed solution;
step 107: dissolving amino beta cyclodextrin in dimethyl sulfoxide to obtain a second mixed solution;
step 108: dropwise adding the second mixed solution into the seventh mixed solution, and stirring for reaction to obtain an eighth mixed solution;
step 109: purifying the eighth mixed solution by a dialysis method to obtain the vitamin A modified cyclodextrin.
Preferably, the vitamin A modified cyclodextrin is prepared by the following steps:
step 102: 2.8g of dicarboxylated polyethylene glycol, 170mg of 4-dimethylaminopyridine and 800mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride were dissolved in 20mL of dimethyl sulfoxide to obtain a fourth mixed solution;
step 103: 364mg of vitamin A is dissolved in 20mL of dimethyl sulfoxide to obtain a fifth mixed solution;
step 104: dropwise adding the fifth mixed solution into the fourth mixed solution under the protection of nitrogen at 25 ℃ and in a dark condition, and stirring and reacting for 48 hours to obtain a sixth mixed solution;
step 105: dialyzing the sixth mixed solution through a 2000Da dialysis bag, and freeze-drying the liquid in the dialysis bag to obtain vitamin A modified polyethylene glycol;
step 106: 988mg of vitamin A modified polyethylene glycol, 153.4mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and 92mg of N-hydroxysuccinimide were dissolved in 20mL of dimethyl sulfoxide to obtain a seventh mixed solution;
step 107: 453.6mg of amino beta cyclodextrin is dissolved in 10mL of dimethyl sulfoxide to obtain a second mixed solution;
step 108: dropwise adding the second mixed solution into the seventh mixed solution at 25 ℃, and stirring and reacting for 72 hours to obtain an eighth mixed solution;
step 109: and (3) performing light-shielding dialysis on the eighth mixed solution for 48 hours through a 3500DA dialysis bag, and performing freeze drying on the liquid in the dialysis bag to obtain the vitamin A modified cyclodextrin.
Preferably, the preparation method of the dicarboxylated polyethylene glycol comprises the following steps:
PEG2000 g was weighed and dissolved completely in 20mL of methylene chloride to obtain PEG 2000-methylene chloride solution;
0.5g of succinic anhydride and 122mg of 4-dimethylaminopyridine are weighed and completely dissolved in 10mL of dichloromethane, and are added into PEG 2000-dichloromethane solution drop by drop under magnetic stirring to react for 12 hours at 60 ℃;
after the reaction is finished, adding deionized water into a reaction system for extraction, hydrating and removing excessive succinic anhydride, collecting an extracted dichloromethane layer, adding color-changing silica gel, drying, and removing dichloromethane by rotary evaporation; and adding 20mL of glacial ethyl ether to form a precipitate, and filtering the precipitate to obtain the dicarboxylated polyethylene glycol after drying the precipitate.
Preferably, the cyclodextrin derivative is used for preparing inclusion compound.
Preferably, the cyclodextrin derivative is used for preparing an inclusion compound of the dihydrotanshinone I;
the clathrate of dihydrotanshinone I is used for preparing medicine or food for resisting hepatic fibrosis.
Preferably, the preparation method of the inclusion compound comprises the following steps:
respectively adding the cyclodextrin derivatives with the dihydrotanshinone I and the triple molar ratio into a mortar, adding a small amount of water and an organic solvent for wetting, grinding for 30 minutes, washing with absolute ethyl alcohol for 1-3 times, collecting an eluate, and drying in vacuum for 12 hours to obtain an inclusion compound of the dihydrotanshinone I; or alternatively
Preparing 4mg/mL cyclodextrin derivative water solution; 10mg of dihydrotanshinone I is weighed and dissolved in 1mL of absolute ethyl alcohol, and then added into a cyclodextrin derivative aqueous solution, and stirring is carried out for 3 days, wherein the molar ratio of the dihydrotanshinone I to the cyclodextrin derivative is 1:3, a step of; after the reaction is finished, filtering by a microporous filter membrane with the diameter of 0.22 mu m, and freeze-drying the filtrate to obtain the clathrate compound of the dihydrotanshinone I.
Compared with the prior art, the invention has the beneficial effects that: the PEG modified cyclodextrin can improve the solubility of the cyclodextrin, improve the in vivo drug absorption quantity and obviously improve the drug distribution quantity of the liver part; the vitamin A modified cyclodextrin ensures that the inclusion compound is easy to be absorbed by hepatic stellate cells, and can improve the hepatic targeting of the inclusion compound, thereby improving the curative effect and reducing the toxic and side effects.
Drawings
FIG. 1 is a flow chart of a process for preparing cyclodextrin derivatives of the present invention;
FIG. 2 is a mass spectrum of a cyclodextrin derivative;
FIG. 3 is a hydrogen spectrum of a cyclodextrin derivative;
FIG. 4 is an infrared spectrum of a cyclodextrin derivative;
FIG. 5 is a scanning electron microscope image of the cyclodextrin derivative-carried dihydrotanshinone I prepared by the grinding method;
FIG. 6 is a scanning electron microscope image of the cyclodextrin derivative loaded with dihydrotanshinone I prepared by the saturated solvent method;
FIG. 7 is a transmission electron microscope image of a cyclodextrin derivative-loaded dihydrotanshinone I dissolved in water;
FIG. 8 is an X-ray powder diffraction pattern of the cyclodextrin derivative-loaded dihydrotanshinone I;
FIG. 9 is a differential scanning calorimetric diagram of a cyclodextrin derivative carrying dihydrotanshinone I;
FIG. 10 is a simulation of RMSD of a cyclodextrin derivative carrying dihydrotanshinone I;
FIG. 11 is a diagram showing the molecular docking of cyclodextrin with dihydrotanshinone I;
FIG. 12 is a graph of liver targeting assays for cyclodextrin derivatives;
FIG. 13 is a graph showing the anti-hepatic fibrosis effect of cyclodextrin derivatives.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The invention is described in further detail below with reference to the attached drawing figures:
a cyclodextrin derivative which is PEG-modified cyclodextrin (PEG-CD) having the structural formula:
the PEG modified cyclodextrin can improve the solubility of the cyclodextrin and the in vivo drug absorption capacity; in one specific test, the drug distribution in the liver region was significantly improved.
Vitamin A can be connected to the tail end of the PEG long chain, namely the cyclodextrin derivative is vitamin A modified cyclodextrin (VA-PEG-CD), and the structural formula is as follows:
its molecular formula can be expressed as [ C ] 20 H 29 OOC(CH 2 ) 2 COO(CHCHO) n OC(CH 2 ) 2 COC 42 H 70 NO 34 ]。
Because vitamin A has a tendency of being easily absorbed by liver cells, the vitamin A modified cyclodextrin formed by grafting the vitamin A on the carboxyl terminal of the PEG modified cyclodextrin has liver targeting, and an inclusion compound formed by a main molecule and a small molecule drug is easily absorbed by hepatic stellate cells (Hepaticstellate cell, HSCs) of the liver, so that the liver targeting of the drug can be improved, the curative effect is improved, and the toxic and side effects are reduced. Among them, HSCs cells are vitamin a (vitamin a, VA) reserve cells, which store about 80% of VA in humans.
The preparation method of the PEG modified cyclodextrin comprises the following steps:
step S1: obtaining the double carboxylated polyethylene glycol.
Step S2: the dicarboxylated polyethylene glycol, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (edc·hcl) and N-hydroxysuccinimide (NHS) were dissolved in dimethyl sulfoxide to obtain a first mixed solution.
Step S3: and dissolving the amino beta cyclodextrin in dimethyl sulfoxide to obtain a second mixed solution.
Step S4: and (3) dropwise adding the second mixed solution into the first mixed solution, and stirring for reaction to obtain a third mixed solution.
Step S5: the third mixture was purified by dialysis to obtain PEG-modified cyclodextrin (PEG-CD).
The preparation method of the vitamin A modified cyclodextrin (VA-PEG-CD) comprises the following steps:
step 101: obtaining the double carboxylated polyethylene glycol.
Step 102: the dicarboxylated polyethylene glycol, 4-dimethylaminopyridine and 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride were dissolved in dimethyl sulfoxide to obtain a fourth mixed solution.
Step 103: vitamin a was dissolved in dimethyl sulfoxide to obtain a fifth mixed solution.
Step 104: and (3) dropwise adding the fifth mixed solution into the fourth mixed solution, and stirring for reaction to obtain a sixth mixed solution.
Step 105: and purifying the sixth mixed solution by a dialysis method to obtain vitamin A modified polyethylene glycol (VA-PEG-COOH).
Step 106: vitamin A modified polyethylene glycol, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide are dissolved in dimethyl sulfoxide to obtain a seventh mixed solution.
Step 107: and dissolving the amino beta cyclodextrin in dimethyl sulfoxide to obtain a second mixed solution.
Step 108: and (3) dropwise adding the second mixed solution into the seventh mixed solution, and stirring for reaction to obtain an eighth mixed solution.
Step 109: purifying the eighth mixed solution by a dialysis method to obtain vitamin A modified cyclodextrin (VA-PEG-CD).
Example 1
PEG-CD and VA-PEG-CD were prepared.
1. Preparation of a dicarboxylated polyethylene glycol (COOH-PEG-COOH):
PEG2000 g was weighed and dissolved completely in 20mL of methylene chloride to obtain PEG 2000-methylene chloride solution; succinic anhydride 0.5g and 4-Dimethylaminopyridine (DMAP) 122mg were weighed, completely dissolved in 10mL of methylene chloride, added dropwise to PEG 2000-methylene chloride solution under magnetic stirring, and reacted at 60℃for 12 hours. After the reaction is finished, adding deionized water into a reaction system for extraction, and hydrating and removing excessive succinic anhydride; adding deionized water for secondary extraction; collecting the extracted dichloromethane layer, adding color-changing silica gel into the dichloromethane layer, drying, and removing dichloromethane by rotary evaporation; and adding 20mL of glacial ethyl ether to form a precipitate, and filtering the precipitate to obtain the dicarboxylated polyethylene glycol (COOH-PEG-COOH) after drying the precipitate.
The structural formula of the dicarboxylated polyethylene glycol can be expressed as:
2. preparation of PEG-modified cyclodextrin (PEG-CD):
step S2: 880mg of dicarboxylated polyethylene glycol, 153.4mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC. HCl) and 92mg of N-hydroxysuccinimide (NHS) were dissolved in 20mL of dimethyl sulfoxide (DMSO) to obtain a first mixture.
Step S3: 453.6mg of amino beta cyclodextrin, which is mono-6-O-amino-beta-cyclodextrin, was dissolved in 10mL of dimethyl sulfoxide to obtain a second mixed solution.
Step S4: the second mixed solution was added dropwise to the first mixed solution at 25℃and reacted with stirring for 72 hours to obtain a third mixed solution.
Step S5: after the third mixed solution was dialyzed for 48 hours by a 3500Da dialysis bag, the liquid in the dialysis bag was freeze-dried to obtain a powdery PEG-modified cyclodextrin (PEG-CD).
3. Preparation of vitamin A modified cyclodextrin (VA-PEG-CD):
step 102: 2.8g of dicarboxylated polyethylene glycol, 170mg of 4-Dimethylaminopyridine (DMAP) and 800mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC. HC l) were dissolved in 20mL of dimethyl sulfoxide (DMSO) to obtain a fourth mixed solution.
Step 103: 364mg of vitamin A was dissolved in 20mL of dimethyl sulfoxide to obtain a fifth mixed solution.
Step 104: and (3) dropwise adding the fifth mixed solution into the fourth mixed solution under the protection of nitrogen at 25 ℃ and in a dark condition, and stirring and reacting for 48 hours to obtain a sixth mixed solution.
Step 105: and (3) dialyzing the sixth mixed solution through a 2000Da dialysis bag, and freeze-drying the liquid in the dialysis bag to obtain the vitamin A modified polyethylene glycol (VA-PEG-COOH).
The structural formula of VA-PEG-COOH can be represented as:
step 106: 988mg of vitamin A modified polyethylene glycol (VA-PEG-COOH), 153.4mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride (EDC. HCL) and 92mg of N-hydroxysuccinimide (NHS) were dissolved in 20mL of dimethyl sulfoxide (DMSO) to obtain a seventh mixed solution.
Step 107: 453.6mg of amino beta cyclodextrin was dissolved in 10mL of dimethyl sulfoxide to obtain a second mixed solution.
Step 108: the second mixed solution was added dropwise to the seventh mixed solution at 25℃and reacted with stirring for 72 hours to obtain an eighth mixed solution.
Step 109: and (3) performing light-shielding dialysis on the eighth mixed solution for 48 hours through a 3500DA dialysis bag, and performing freeze drying on the liquid in the dialysis bag to obtain the vitamin A modified cyclodextrin (VA-PEG-CD).
Subjecting prepared COOH-PEG-COOH, VA-PEG-COOH and VA-PEG-CD to Mass Spectrometry (MS) and nuclear magnetic resonance hydrogen spectrometry respectively 1 H-NMR) and infrared spectroscopy (I R). Mass spectrum characterization results are shown in fig. 2, wherein the charges of COOH-PEG-COOH, VA-PEG-COOH and VA-PEG-CD are 1, the average mass-to-charge ratio/average molecular weight of the dicarboxylated polyethylene glycol (COOH-PEG-COOH) is 2200, the average mass-to-charge ratio/average molecular weight of VA-PEG-COOH is 2400, and the average mass-to-charge ratio/average molecular weight of VA-PEG-CD is 3500. The nuclear magnetic resonance hydrogen spectrum characterization result is shown in FIG. 3, wherein 3.52ppm of characteristic peak is attributed to methylene in the PEG repeating unit, and 2.50ppm of characteristic peak is attributed to methylene in succinic anhydride end group reacted with PEG; 1.07ppm of characteristic peak is assigned to grafted VA methyl peak, and 2.13ppm is methyl peak on the double bond structure of VA and PEG connection; 4.83ppm Cyclodextrin H-1Peaks, 3.34 and 3.64ppm are H-2 and H-6 peaks, respectively, in the cyclodextrin derivative, and 5.70ppm is OH peak of the cyclodextrin derivative, and according to the above nuclear magnetic resonance hydrogen spectrum results, it can be confirmed that the dicarboxylated PEG molecule was successfully linked to VA and cyclodextrin, respectively. The infrared spectrum characterization result is shown in figure 4, 2800-3000 cm -1 C-H stretching vibration peaks of methyl and methylene; 1730cm in VA-PEG-COOH pattern compared to COOH-PEG-COOH pattern -1 Is reduced in absorption of C=O, and 1631cm -1 C=C stretching vibration peak of VA is added, which proves that VA is successfully grafted to COOH-PEG-COOH; VA-PEG-CD was at 3418cm compared to VA-PEG-COOH infrared spectrum -1 The absorption of the stretching vibration peak of the hydroxyl group is enhanced, which indicates that the cyclodextrin is successfully grafted with VA-PEG-COOH.
Example 2
Preparation of dihydrotanshinone (DH I) clathrate (CD-DH I, PEG-CD-DH I and VA-PEG-CD-DH I). Preparing DH I inclusion compound by grinding method and saturated solvent method respectively:
preparing inclusion compound by grinding method:
adding DH I and cyclodextrin derivatives with triple molar ratio into mortar, adding a small amount of water and organic solvent for wetting, grinding for 30 min, washing with absolute ethanol for 1-3 times, collecting eluate, and vacuum drying for 12h to obtain cyclodextrin derivative DH I clathrate.
The cyclodextrin carried dihydrotanshinone clathrate (CD-DH I) and the PEG modified cyclodextrin carried dihydrotanshinone clathrate (PEG-CD-DH I) and the VA modified cyclodextrin carried dihydrotanshinone clathrate (VA-PEG-CD-DH I) are respectively prepared by adopting the method.
Preparing inclusion compound by saturated solvent method:
preparing 4mg/mL cyclodextrin derivative water solution; 10mg of dihydrotanshinone I is weighed and dissolved in 1mL of absolute ethyl alcohol, and then added into a cyclodextrin derivative aqueous solution, and stirring is carried out for 3 days, wherein the molar ratio of the dihydrotanshinone I to the cyclodextrin derivative is 1:3, a step of; after the reaction is finished, filtering by a microporous filter membrane with the diameter of 0.22 mu m, and freeze-drying the filtrate to obtain the clathrate compound of the dihydrotanshinone I.
The cyclodextrin derivative loaded with the dihydrotanshinone I inclusion compound prepared by the method is subjected to Scanning Electron Microscope (SEM), transmission Electron Microscope (TEM), differential Scanning Calorimetry (DSC) and X-ray powder diffraction (XRD) measurement. The scanning electron microscope results are shown in fig. 5 and 6, the inclusion compound prepared by the grinding method presents random blocks, and the inclusion compound prepared by the saturated solvent method presents a sheet shape after freeze-drying; the transmission electron microscopy results are shown in fig. 7, and compared with the unmodified cyclodextrin inclusion compound, the VA modified cyclodextrin-loaded dihydrotanshinone inclusion compound (dissolved in water) presents a more regular spheroid shape.
XRD and DSC results are shown in fig. 8 and 9, respectively. DSC shows that DH I has a strong endothermic peak at 230 ℃ from figure 9; from FIG. 8, DH I has a strong characteristic peak; and the cyclodextrin derivative carries DH I to form inclusion compound with the endothermic peak and the characteristic peak disappeared, which indicates that DH I does not exist in the crystal form after grinding.
The compound and receptor modes of action were analyzed using an autodock4.2 calculation to obtain the compound and receptor actions, such as hydrogen bonding, pi stacking, hydrophobic interactions, etc., and stability of the complex formed was estimated by referencing the compound's docking score. The molecular docking results are shown in table 1.
TABLE 1
In order to further study the interactions of small molecules with cyclodextrin, molecular dynamics simulations of cyclodextrin and small molecule complexes were performed for 20ns using molecular dynamics. The Root Mean Square Deviation (RMSD) of cyclodextrin and small molecule complex was monitored to allow for deep knowledge of the structural conformational changes throughout the simulation. RMSD analysis may indicate whether the simulation has equilibrated; the RMSD value is typically stable around a fixed value. If the RMSD of the cyclodextrin is still increasing or decreasing on average at the end of the simulation, the system is not yet balanced and the simulation time may be insufficient for a rigorous analysis. Likewise, RMSD also reflects the stability of cyclodextrin with small molecules, with larger RMSDs indicating that this cyclodextrin system is more unstable.
Molecular docking results show that the compound DHI has good binding effect with target receptorsAnd the matching degree is high, and the binding energy is-5.78 kcal/mol. The compound formed by the compound and the receptor after the butt joint is visually analyzed by using the Pymol2.1 software to obtain the combination mode of the compound and the receptor, three hydrogen bond interactions between DHI and CD can be clearly seen according to the combination mode, and the hydrogen bond distances are respectively
Far less than the traditional hydrogen bond +.>
The strong binding capacity is an important contribution to promoting the formation of stable complexes between the two molecules. In addition, because DHI contains a plurality of rings, the DHI has certain hydrophobicity, can form good hydrophobic interaction with a hydrophobic cavity of cyclodextrin, and also has certain contribution to stabilizing small molecules.
As shown in FIG. 10, the average RMSD of cyclodextrins is approximately
And the complex achieves dynamic balance in a short time, which indicates that small molecule DHI is well matched with cyclodextrin targets and can form stable complex with cyclodextrin. In addition, as can be seen from the results in the figure, the conformational change of the complex does not present a significant fault problem, which also indicates that small molecule DHI can bind well to cyclodextrin without leaving the active pocket of cyclodextrin. As can be seen from fig. 11, the small molecules did not leave the cyclodextrin cavity throughout the simulation, and further analysis of the 5ns (fig. 11A), 10ns (fig. 11B), 15ns (fig. 11C) and 20ns (fig. 11D) conformations revealed that these compounds were able to form hydrogen bonds, conjugation, hydrophobic interactions with the cyclodextrin cavity, and these interactions were effective to promote the formation of stable complexes of small molecule DHI with cyclodextrin. In conclusion, DHI and cyclodextrin derivatives can form a variety of inclusion compounds with high interaction, matching degree and stability.
Liver targeting assay for cyclodextrin derivatives
Preparing a cyclodextrin derivative-carried nile red clathrate compound: and (3) replacing DHI with nile red to prepare the cyclodextrin derivative nile red inclusion compound.
Animal BDL model establishment: SD male rats (weight is 180-200 g) are taken as test animals, the test animals are fasted for 12 hours before operation, after anesthesia by isoflurane, under the aseptic operation condition, the abdomen is opened, the liver margin is lifted, the duodenum is pulled open, the common bile duct is separated by 2-3cm, the near duodenum and the near hepatic portal are respectively ligated by using No. 000 silk threads for two times, the common bile duct is cut off from the middle of the two ligation positions, and the incision is sutured after the liver is recovered. After the animals were anesthetized and awake, they were normally fed with water and were allowed to drink water freely, thereby obtaining BDL rats.
BDL rats are respectively injected with nile red (free), cyclodextrin-carried nile red inclusion compound and PEG-modified cyclodextrin-carried nile red inclusion compound at doses of 2mg/kg in an intraperitoneal mode, after injection is finished, living body imaging analysis is carried out on the rats, then chloral hydrate solution is injected to kill the animals, and liver tissues of the animals are collected and subjected to imaging analysis. As shown in fig. 12A and 12B, the fluorescence intensity/average emissivity of the PEG-modified cyclodextrin-carried nile red clathrate compound is obviously higher than that of the cyclodextrin-carried nile Luo Gongbao compound and free nile red in 8 hours, and thus, the hepatic tissue drug distribution amount of the clathrate compound can be improved by PEG-modified cyclodextrin; as can be seen in fig. 12A, 12B and 12C, nile red accumulation in the liver was further enhanced following the VA modified cyclodextrin loading of nile red.
Anti-hepatic fibrosis drug efficacy detection
The BDL rats are randomly taken as a BDL control group, a dihydrotanshinone administration group, a cyclodextrin loaded dihydrotanshinone administration group, a PEG modified cyclodextrin loaded dihydrotanshinone administration group and a VA modified cyclodextrin loaded dihydrotanshinone administration group. The groups were administered by intraperitoneal injection (dosage of dihydrotanshinone is 10 mg/kg) on the 2 nd day after the operation, 1 time a day, and after 14 days of administration, samples such as blood, bile, liver, etc. were collected after 12 hours of fasting; the BDL control group was injected with physiological saline intraperitoneally at the 2 nd day after the operation, 1 time a day, 14 days after the administration of physiological saline, and after 12 hours of fasting, samples of blood, bile, liver, etc. were collected.
Group of sham operations: fasted for 12 hours before the operation of the experimental animal; surgery: after anesthesia with isoflurane, opening the abdomen under aseptic operation condition, then suturing the incision, and after the animals are anesthetized and awake, eating normally and drinking water freely; physiological saline is injected into the abdominal cavity on the 2 nd day after the operation, 1 time a day, 14 days later, and after fasting for 12 hours, samples such as blood, bile, liver and the like are collected.
Hematoxylin-eosin, sirius red and masson staining were performed on liver tissue sections, respectively, and the results are shown in fig. 13, and compared with the sham operation group, bile duct proliferation and necrosis of the BDL control group were significantly increased; after the dihydrotanshinone and the cyclodextrin derivative inclusion compound are administrated, the VA-modified cyclodextrin inclusion compound can obviously improve the hyperplasia and necrosis of the liver bile duct. The staining results of sirius red and masson can reflect the collagen deposition condition in the liver, and the more collagen deposition proves the more serious the liver fibrosis degree. The BDL control group can see serious collagen deposition, and after PEG modified cyclodextrin inclusion compound is administrated, the collagen deposition is reduced; after the VA modified cyclodextrin inclusion compound is administrated, collagen deposition is obviously reduced, which indicates that the VA modified cyclodextrin inclusion compound can obviously improve liver fibrosis degree of BDL rats. Therefore, the VA modified cyclodextrin is used as a carrier, and the prepared clathrate compound of the dihydrotanshinone I can be used for preparing medicines or foods for resisting hepatic fibrosis.
The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A cyclodextrin derivative, characterized in that the cyclodextrin derivative is PEG modified cyclodextrin or vitamin A modified cyclodextrin,
wherein, the PEG modified cyclodextrin has the structural formula:
the vitamin A modified cyclodextrin has the structural formula:
2. the method for preparing cyclodextrin derivatives according to claim 1, wherein the method for preparing PEG-modified cyclodextrin comprises:
step S1: obtaining dicarboxylated polyethylene glycol;
step S2: dissolving dicarboxylated polyethylene glycol, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide in dimethyl sulfoxide to obtain a first mixed solution;
step S3: dissolving amino beta cyclodextrin in dimethyl sulfoxide to obtain a second mixed solution;
step S4: dropwise adding the second mixed solution into the first mixed solution, and stirring for reaction to obtain a third mixed solution;
step S5: and purifying the third mixed solution by a dialysis method to obtain the PEG modified cyclodextrin.
3. The method of claim 2, wherein the method of preparing PEG-modified cyclodextrin comprises:
step S2: 880mg of dicarboxylated polyethylene glycol, 153.4mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and 92mg of N-hydroxysuccinimide were dissolved in 20mL of dimethyl sulfoxide to obtain a first mixed solution;
step S3: 453.6mg of amino beta cyclodextrin is dissolved in 10mL of dimethyl sulfoxide to obtain a second mixed solution, wherein the amino beta cyclodextrin is mono-6-O-amino-beta cyclodextrin;
step S4: dropwise adding the second mixed solution into the first mixed solution at 25 ℃, and stirring and reacting for 72 hours to obtain a third mixed solution;
step S5: and (3) dialyzing the third mixed solution for 48 hours through a 3500DA dialysis bag, and freeze-drying the liquid in the dialysis bag to obtain the PEG modified cyclodextrin.
4. The method of preparing vitamin a modified cyclodextrin according to claim 2, comprising:
step 101: obtaining dicarboxylated polyethylene glycol;
step 102: dissolving dicarboxylated polyethylene glycol, 4-dimethylaminopyridine and 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride in dimethyl sulfoxide to obtain a fourth mixed solution;
step 103: dissolving vitamin A in dimethyl sulfoxide to obtain a fifth mixed solution;
step 104: dropwise adding the fifth mixed solution into the fourth mixed solution, and stirring for reaction to obtain a sixth mixed solution;
step 105: purifying the sixth mixed solution by a dialysis method to obtain vitamin A modified polyethylene glycol;
step 106: dissolving vitamin A modified polyethylene glycol, 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and N-hydroxysuccinimide in dimethyl sulfoxide to obtain a seventh mixed solution;
step 107: dissolving amino beta cyclodextrin in dimethyl sulfoxide to obtain a second mixed solution;
step 108: dropwise adding the second mixed solution into the seventh mixed solution, and stirring for reaction to obtain an eighth mixed solution;
step 109: purifying the eighth mixed solution by a dialysis method to obtain the vitamin A modified cyclodextrin.
5. The method of claim 4, wherein the method of preparing vitamin a modified cyclodextrin comprises:
step 102: 2.8g of dicarboxylated polyethylene glycol, 170mg of 4-dimethylaminopyridine and 800mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride were dissolved in 20mL of dimethyl sulfoxide to obtain a fourth mixed solution;
step 103: 364mg of vitamin A is dissolved in 20mL of dimethyl sulfoxide to obtain a fifth mixed solution;
step 104: dropwise adding the fifth mixed solution into the fourth mixed solution under the protection of nitrogen at 25 ℃ and in a dark condition, and stirring and reacting for 48 hours to obtain a sixth mixed solution;
step 105: dialyzing the sixth mixed solution through a 2000Da dialysis bag, and freeze-drying the liquid in the dialysis bag to obtain vitamin A modified polyethylene glycol;
step 106: 988mg of vitamin A modified polyethylene glycol, 153.4mg of 1-ethyl- (3-dimethylaminopropyl) carbodiimide hydrochloride and 92mg of N-hydroxysuccinimide were dissolved in 20mL of dimethyl sulfoxide to obtain a seventh mixed solution;
step 107: 453.6mg of amino beta cyclodextrin is dissolved in 10mL of dimethyl sulfoxide to obtain a second mixed solution;
step 108: dropwise adding the second mixed solution into the seventh mixed solution at 25 ℃, and stirring and reacting for 72 hours to obtain an eighth mixed solution;
step 109: and (3) performing light-shielding dialysis on the eighth mixed solution for 48 hours through a 3500DA dialysis bag, and performing freeze drying on the liquid in the dialysis bag to obtain the vitamin A modified cyclodextrin.
6. The method according to claim 2 or 4, wherein the method for preparing the dicarboxylated polyethylene glycol comprises:
PEG2000 g was weighed and dissolved in 20mL of methylene chloride to obtain PEG 2000-methylene chloride solution;
0.5g of succinic anhydride and 122mg of 4-dimethylaminopyridine are weighed, dissolved in 10mL of dichloromethane, added dropwise into PEG 2000-dichloromethane solution under magnetic stirring, and reacted for 12 hours at 60 ℃;
after the reaction is finished, adding deionized water into a reaction system for extraction, hydrating and removing excessive succinic anhydride, collecting an extracted dichloromethane layer, adding color-changing silica gel, drying, and removing dichloromethane by rotary evaporation; and adding 20mL of glacial ethyl ether to form a precipitate, and filtering the precipitate to obtain the dicarboxylated polyethylene glycol after drying the precipitate.
7. Use of a cyclodextrin derivative according to claim 1 for the preparation of an inclusion compound.
8. The use according to claim 7, wherein the cyclodextrin derivative is used for the preparation of clathrates of dihydrotanshinone i.
9. The use according to claim 8, wherein the clathrate of dihydrotanshinone i is used for preparing anti-hepatic fibrosis medicine or food.
10. The use according to claim 8, wherein the process for the preparation of the inclusion compound comprises:
respectively adding the cyclodextrin derivatives with the dihydrotanshinone I and the triple molar ratio into a mortar, adding a small amount of water and an organic solvent for wetting, grinding for 30 minutes, washing with absolute ethyl alcohol for 1-3 times, collecting washing liquid, and drying in vacuum for 12 hours to obtain an inclusion compound of the dihydrotanshinone I; or alternatively
Preparing 4mg/mL cyclodextrin derivative water solution; 10mg of dihydrotanshinone I is weighed and dissolved in 1mL of absolute ethyl alcohol, and then added into a cyclodextrin derivative aqueous solution, and stirring is carried out for 3 days, wherein the molar ratio of the dihydrotanshinone I to the cyclodextrin derivative is 1:3, a step of; after the reaction is finished, filtering by a microporous filter membrane with the diameter of 0.22 mu m, and freeze-drying the filtrate to obtain the clathrate compound of the dihydrotanshinone I.
Priority Applications (1)
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| CN202310019403.XA CN116144031A (en) | 2023-01-06 | 2023-01-06 | Cyclodextrin derivatives, and preparation method and application thereof |
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| CN202310019403.XA CN116144031A (en) | 2023-01-06 | 2023-01-06 | Cyclodextrin derivatives, and preparation method and application thereof |
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