CN108030768A - The preparation of the different efficient cross-film polymer micelle of molecular modification type and its pharmaceutical applications - Google Patents

The preparation of the different efficient cross-film polymer micelle of molecular modification type and its pharmaceutical applications Download PDF

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CN108030768A
CN108030768A CN201711343697.2A CN201711343697A CN108030768A CN 108030768 A CN108030768 A CN 108030768A CN 201711343697 A CN201711343697 A CN 201711343697A CN 108030768 A CN108030768 A CN 108030768A
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micelle
carrier
efficient cross
film
polymer micelle
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殷婷婕
霍美蓉
包聪聪
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China Pharmaceutical University
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China Pharmaceutical University
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
    • A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • 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
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
    • 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
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/34—Macromolecular compounds obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyesters, polyamino acids, polysiloxanes, polyphosphazines, copolymers of polyalkylene glycol or poloxamers
    • 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
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
    • C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
    • C08B37/0027—2-Acetamido-2-deoxy-beta-glucans; Derivatives thereof
    • C08B37/003—Chitin, i.e. 2-acetamido-2-deoxy-(beta-1,4)-D-glucan or N-acetyl-beta-1,4-D-glucosamine; Chitosan, i.e. deacetylated product of chitin or (beta-1,4)-D-glucosamine; Derivatives thereof
    • C—CHEMISTRY; METALLURGY
    • C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
    • C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
    • C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
    • C08B37/006—Heteroglycans, i.e. polysaccharides having more than one sugar residue in the main chain in either alternating or less regular sequence; Gellans; Succinoglycans; Arabinogalactans; Tragacanth or gum tragacanth or traganth from Astragalus; Gum Karaya from Sterculia urens; Gum Ghatti from Anogeissus latifolia; Derivatives thereof
    • C08B37/0063—Glycosaminoglycans or mucopolysaccharides, e.g. keratan sulfate; Derivatives thereof, e.g. fucoidan
    • C08B37/0072—Hyaluronic acid, i.e. HA or hyaluronan; Derivatives thereof, e.g. crosslinked hyaluronic acid (hylan) or hyaluronates
    • 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

Abstract

Preparation and its pharmaceutical applications the present invention relates to the efficient cross-film polymer micelle of different molecular modification type, the micellar carrier is by first introducing the hydrophobic molecule for having efficient cross-film function on hydrophilic macromolecule chain backbone, and the hydrophobic molecule for further introducing the sensitivity key of microenvironment containing lesion builds to obtain.Destination carrier have it is amphipathic, and in an aqueous medium can self assembly be nano-micelle, its kernel can load hydrophobic drug.It is primarily characterized in that：1) hydrophobic molecule of the efficient cross-film function of tool in micellar structure can mediate the efficient cross-film of nano-micelle to enter born of the same parents, promote more carrier micelles by lesion cell endocytic；2) the lesion microenvironment sensitivity key contained by micellar structure can be by focal zone microenvironment selective degradation, cause carrier micelle hydrophobic inner core reaction force attenuation, particle diameter becomes larger, and insoluble drug release accelerates to improve lesions position drug concentration, improves drug bioavailability and its corresponding pharmacological activity effect.

Description

The preparation of the different efficient cross-film polymer micelle of molecular modification type and its pharmaceutical applications
Technical field
The invention belongs to field of pharmaceutical preparations.It is related to a kind of efficient cross-film polymer micelle of different molecular modification type as medicine Delivery vector, the invention further relates to the preparation method and its pharmaceutical applications of the polymer micelle.
Background technology
Hydrophobic drug has that bioavilability is low, oral absorption is poor, and the shortcomings of be difficult to prepare suitable formulations.Polymerization Thing micella comes into being as the means of medicine delivery.Amphipathic nature block polymer can be self-assembly of inner casing in aqueous Nucleocapsid structure hydrophobic, shell is hydrophilic, i.e. polymer micelle, are a kind of novel nano medicine-carried systems fast-developing in recent years, Go out as drug carriers display than other nano-carriers more vast potential for future development., can be to dredging since inner casing has hydrophobicity Aqueous pharmaceutical is wrapped up, and increases the solubility and stability of medicine.Meanwhile polymer micelle particle diameter I passes through high-permeability With retention effect (enhanced permeability and retention effect, EPR effect) increase lesions position Accumulation, it is safe, also with low critical micelle concentration (critical micelle concentration, CMC) and heating power Learn the advantages that stability is good.
But means of the current most polymers micella as delivering medicine, also fail to solve problems with：(1) many conjunctions Into high molecular material, there are the defects of a degree of haemolysis, heat source response and permeability, poor biocompatibility etc..(2) Carrier micelle is delivered to lesions position, has the hydrophilic outer shell meeting block cell encytosis compared with strongly hydrophilic, reduces phagocytosis effect Rate, reduces drug effect.(3) general polymer micella relies on the slow of polymer in lesions position without special response, insoluble drug release Degraded, therefore it is slow to release the drug.
Carrier micelle can enter lesion cell by endocytosis.It is modified with the hydrophobic molecule for having efficient cross-film function Polymer medicament carrying micelle, born of the same parents more can be entered by lesion cell endocytic, efficient cross-film, improve drug effect.And participate in forming cell The cholesterol molecule of membrane structure has good biomembrane compatibility and stronger hydrophobicity, thus cholesterol or derivatives thereof point Son has the function of efficient cross-film.
Focal zone generally has special microenvironment, such as high reducing environment, acidity, active oxygen (Reactive oxygen Species, ROS), weary oxygen etc., can the efficient corresponding microenvironment sensitivity key of differential stimulus, such as reduce sensitive disulfide bond；It is acid-sensitive Feel key：Phenyl boric acid ester bond, acetal bonds, hydrazone key；ROS sensitivity keys：Boric acid ester bond, thio ketal ization key, thioether bond；Anoxic sensitivity azo Key.The polymer medicament carrying micelle for having such sensitive key hydrophobic molecule is modified with after lesions position is reached, can specificly-response disease Stove microenvironment causes micellar structure to destroy, and realizes quick drug release.
But only to have polymer micelle and the unresolved biography that the hydrophobic molecule of efficient cross-film function is built as hydrophobic grouping The shortcomings that polymer micelle drug release of uniting is slow, and only built by hydrophobic grouping of the hydrophobic molecule containing lesion microenvironment sensitivity key Polymer micelle, and the shortcomings that unresolved traditional polymer micella enters born of the same parents' efficiency, it is also possible to have into born of the same parents' efficiency it is low, stimulate and ring Should after the shortcomings that causing the sedimentation of hydrophobic ingredient rapid property to cause drug molecule can not be scattered in microenvironment with molecular forms.At present not yet Report has the polymer latex of the double modification types of hydrophobic molecule of the hydrophobic molecule and the sensitivity key of microenvironment containing lesion of efficient cross-film function Beam and pharmaceutical composition, i.e., the efficient cross-film polymer micelle of different molecular modification type and pharmaceutical composition.
The content of the invention
The purpose of the present invention is for above-mentioned technical problem, there is provided a kind of different efficient cross-film polymer micelle of molecular modification type As drug delivery vehicle.The polymer micelle not only has the effect of efficient cross-film, and contained lesion microenvironment sensitivity key is in disease By after selective degradation, micellar hydrophobic area reaction force attenuation, shows as particle diameter and becomes larger, can accelerate the release of medicine stove microenvironment, Significantly improve drug effect.
It is a further object to provide the preparation method of above-mentioned polymer micelle.
It is a still further object of the present invention to provide application of the above-mentioned polymer micelle in pharmacy.
The inventive concept of the present invention be using hydrophilic macromolecule chain as skeleton, by amido link or ester bond introduce tool efficiently across The hydrophobic molecule of film function, then further introduced by amido link or ester bond to hydrophilic macromolecule chain backbone and contain lesion micro-loop The hydrophobic molecule of border sensitivity key, prepares polymer micelle carrier；The polymer micelle carrier have it is amphipathic, can be in aqueous medium Middle self assembly is nano-micelle, hydrophobic drug molecule can be wrapped in micelle inner core by noncovalent interaction and delivered it to Lesions position；The hydrophobic molecule of the efficient cross-film function of tool in micellar structure, can mediate the efficient cross-film of nano-micelle to enter born of the same parents, promote More carrier micelles are by lesion cell endocytic；In addition, the lesion microenvironment sensitivity key contained by micellar structure can be micro- by focal zone Environment selective degradation, causes nano-micelle kernel hydrophobic forces to weaken, particle diameter increase, medicine, which is fast released, acts on disease Stove area, is remarkably improved the concentration, curative effect and bioavilability of lesions position free drug.
To reach above-mentioned purpose, the present invention provides a kind of different efficient cross-film polymer micelle carrier of molecular modification type, it is tied Structure general formula is as follows：
[R1]q-M-[R2]p
Wherein M is hydrophilic macromolecule chain；R1To have the hydrophobic molecule of efficient cross-film function；R2It is quick for microenvironment containing lesion Feel the hydrophobic molecule of key；Q be hydrophilic macromolecule chain backbone on have efficient cross-film function hydrophobic molecule substitution value, the tool The substitution value of the hydrophobic molecule of efficient cross-film function is 2%~25%；P is microenvironment containing lesion on hydrophilic macromolecule chain backbone The substitution value of the hydrophobic molecule of sensitive key, the substitution value of the hydrophobic molecule of the sensitivity of microenvironment containing the lesion key for 2%~ 70%.
The efficient cross-film polymer micelle carrier of different molecular modification type, wherein hydrophilic macromolecule chain are selected from hyalomitome Acid, unfraction heparin, low molecular weight heparin, desulfated heparin, chondroitin, poly-sulfated chondroitin, alginic acid, glucan, In fungi polysaccharide, chitosan, lentinan, polymaleic anhydride, polyacrylic acid and derivative containing carboxyl, amino or hydroxyl One kind.
The efficient cross-film polymer micelle carrier of different molecular modification type, wherein having the hydrophobic molecule of efficient cross-film function One kind in cholesterol, cholesterol derivative, wherein cholesterol derivative be selected from butanedioic acid cholesterol monoesters, 3 beta-aminos- One kind in 5- cholestene, phthalic acid cholesterol hydrogen ester.
The efficient cross-film polymer micelle carrier of different molecular modification type, wherein the sensitivity key of microenvironment containing lesion is hydrophobic Molecule is selected from lipoic acid, 4- hydroxy benzenes pinacol borate, 4,4 '-dicarboxyl azobenzene, azobenzene -4 benzoic acid, poly- L- groups One kind in propylhomoserin, oligomerization methionine.
The preparation method of the efficient cross-film polymer micelle carrier of different molecular modification type, it is characterised in that including following Step：
(1) the hydrophilic macromolecule chain containing amino, carboxyl, hydroxyl is dissolved in reaction dissolvent, with 1- (3- dimethylaminos third Base) -3- ethyl-carbodiimide hydrochlorides (EDC) and n-hydroxysuccinimide (NHS) or 1- (3- dimethylamino-propyls) -3- second Base carbodiimide hydrochloride (EDC), n-hydroxysuccinimide (NHS) and 4-dimethylaminopyridine (DMAP) or 1- (3- diformazans Aminopropyl) -3- ethyl-carbodiimide hydrochlorides (EDC) and I-hydroxybenzotriazole (HOBT) or 1- (3- dimethylaminos third Base) -3- ethyl-carbodiimide hydrochlorides (EDC) and triethylamine (TEA), 4-dimethylaminopyridine (DMAP) be activator, will have The hydrophobic molecule of efficient cross-film function obtains intermediate 1 by amido link or linkage in hydrophilic macromolecule；
(2) intermediate 1 is dissolved in reaction dissolvent with 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides (EDC) and n-hydroxysuccinimide (NHS) or 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides (EDC), N- HOSu NHS (NHS) and 4-dimethylaminopyridine (DMAP) or 1- (3- dimethylamino-propyls) -3- ethyl carbodiimides Hydrochloride (EDC) and I-hydroxybenzotriazole (HOBT) or 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides (EDC) and triethylamine (TEA), 4-dimethylaminopyridine (DMAP) are activator, by hydrophobic point of the sensitivity key of microenvironment containing lesion Son in intermediate 1, that is, obtains the efficient cross-film polymer micelle carrier of different molecular modification type by amido link or linkage.
The efficient cross-film polymer micelle carrier of different molecular modification type, in preparation process reaction dissolvent be water, methanol, N,N-Dimethylformamide, tetrahydrofuran, toluene, dimethyl sulfoxide (DMSO), dichloromethane, formamide, water and methanol mixed solvent, The mixed solvent of the water and mixed solvent of n,N-Dimethylformamide, water and formamide, n,N-Dimethylformamide and formamide Mixed solvent.
The efficient cross-film polymer micelle carrier of different molecular modification type have it is amphipathic, can spontaneous shape in aqueous medium Into nano-micelle.
The efficient cross-film polymer micelle of different molecular modification type, can be used for the injection of intravascular or intramuscular or mouth Clothes, the carrier with pharmaceutical active or pharmacological activity molecule of external application.Wherein it is selected from pharmaceutical active or pharmacological activity molecule： Non-steroid anti-inflammatory drug, steroidal anti-inflammatory drugs, suppress uric acid generation class anti-gout drugs, promote uric acid discharge class anti-gout drugs And taxanes, flavonoids, camptothecin, vinca, Anthraquinones, cyclosporine class, antifol, purine antagonist, Pyrimidine antagonists, any one in dihydropyridine series antineoplastic medicament or two compositions.
The efficient cross-film polymer micelle of different molecular modification type and the preparation method of composition step of medicine are as follows：
The efficient cross-film polymer micelle carrier of different molecular modification type and water are dissolved by weight for 1~50: 1000 ratio, Through ultrasonic or high-pressure homogeneous processing, nano micellar solution is obtained.By therapeutically effective amount with pharmaceutical active or pharmacological activity point After son is dissolved with pharmaceutically acceptable organic solvent, preformed solution is mixed to get with the nano micellar solution；Will be prefabricated molten Liquid is after ultrasonic or high-pressure homogeneous processing, and using dialysis, ultrafiltration or evaporating organic solvent, it is 10~1000nm to obtain particle diameter Polypeptide drug-loaded micelle solution；Or the rotated evaporation of preformed solution is redissolved or scattered, through ultrasound after bottle wall forms film with water Or high-pressure homogeneous processing, the polypeptide drug-loaded micelle solution that particle diameter is 10~1000nm is prepared.The polypeptide drug-loaded micelle solution being prepared Freeze-dried powder can be made using freeze-drying by adding pharmaceutically acceptable freeze drying protectant.
The pharmaceutically acceptable organic solvent is：Methanol, ethanol, acetone, tetrahydrofuran, N, N- dimethyl formyls The organic solvents such as amine, dimethyl sulfoxide (DMSO) or their mixed solvent.
The polypeptide drug-loaded micelle solution that the efficient cross-film polymer micelle of different molecular modification type is prepared is using addition Freeze drying protectant for lactose, glucose, mannitol, sodium chloride, amino acid, citrate, acetate, one kind in phosphate or It is a variety of.
Beneficial effects of the present invention：
First, the present invention using containing amino, carboxyl, hydroxyl high molecular weight hydrophilic chain as skeleton, pass through amido link or ester bond and introduce Has the hydrophobic molecule of efficient cross-film function；Microenvironment containing lesion is further introduced to macromolecular scaffold by amido link or ester bond again The hydrophobic molecule of sensitive key, constructs the efficient cross-film polymer micelle of different molecular modification type and its pharmaceutical composition；Wherein, tool is efficient The hydrophobic molecule of cross-film function can promote more carrier micelles by lesion cell endocytic；Contained lesion microenvironment sensitivity key is in disease Stove site specific is degraded, and causes hydrophobic region reaction force attenuation, carrier micelle particle diameter to become larger, and accelerates to avoid bag while drug release The medicine wrapped up in the carrier fails to discharge, fails to play the shortcomings that drug effect is eliminated, and is remarkably improved bioavilability and medicine Effect.
2nd, the efficient cross-film polymer micelle of different molecular modification type provided by the invention has good biocompatibility and stabilization Property, also there is the advantage for the seizure for hiding organism reticuloendothelial system, there is actively or passively targeting.
3rd, the efficient cross-film polymer micelle carrier of different molecular modification type provided by the invention has well amphipathic, can Nano-micelle is spontaneously formed in water, effectively can singly be carried or be carried poorly water soluble drugs altogether.
4th, the efficient cross-film polymer micelle of different molecular modification type provided by the invention can be used for intravascular or intramuscular to note Penetrate or take orally, topical route administration.Micella uniform particle sizes degree is good, can be controlled in 10~1000nm, is widely used.
Brief description of the drawings
Fig. 1 is chitosan-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle that embodiment 22 contains taxol (GCT/PTX) and contain taxol chitosan-lipoic acid polymer medicament carrying micelle (GT/PTX) In-vitro release curves.
Embodiment
It is subject to further instruction to the present invention below by embodiment, but following embodiments are not intended to limit the power of this patent Sharp scope.
Embodiment 1：The preparation of hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (HCT)
1.000g hyaluronic acids, 0.080g cholesteryl hemisuccinates are dissolved in formamide and N, N- dimethyl formyls respectively In amine mixed solvent (v/v=1: 1), under condition of ice bath, 0.069g EDC, 0.030g NHS, 0.008g DMAP activation courages are added After 0.5~1h of free carboxy of sterol monomester succinate, it is slowly dropped into the solution of hyaluronic acid, normal-temperature reaction 24h.Used The cold acetone precipitation of amount, filters and purifies sediment, redissolved with water, distilled water dialysis (MWCO=3500) 48~72h.Freezing is dry It is dry to obtain intermediate 1.
0.300g intermediates 1,0.220g lipoic acids are dissolved in formamide and n,N-Dimethylformamide mixed solvent respectively In (v/v=1: 1), under condition of ice bath, add 0.990g EDC, 0.589g NHS, 0.012g DMAP and activate the free of lipoic acid 0.5~1h of carboxyl, is slowly dropped into the solution of intermediate 1, normal-temperature reaction 24h.Using excessive cold acetone precipitation, filter and pure Change sediment, redissolved with water, distilled water dialysis (MWCO=3500) 48~72h.Freeze-drying obtains hyaluronic acid-cholesterol Monomester succinate/lipoic acid polymer micelle carrier (HCT).
Embodiment 2：The preparation of chitosan-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (GCT)
1.000g chitosans, 0.090g cholesteryl hemisuccinates are dissolved in dimethyl sulfoxide (DMSO) respectively, under condition of ice bath, 0.077g EDC, 0.052g NHS, 0.5~1h of free carboxy of 0.009g DMAP activation cholesterol monomester succinates are added, is delayed It is slow to instill in the solution of chitosan, normal-temperature reaction 24h.Distilled water dialysis (MWCO=3500) 48~72h.Freeze-drying obtains Intermediate 1.
0.400g intermediates 1,0.220g lipoic acids are dissolved in dimethyl sulfoxide (DMSO) respectively, under condition of ice bath, added 0.990g EDC, 0.589g NHS, 0.5~1h of free carboxy of 0.015g DMAP activation lipoic acids, are slowly dropped into intermediate 1 Solution in, normal-temperature reaction 24h.Distilled water dialysis (MWCO=3500) 48~72h.It is solid that freeze-drying obtains chitosan-courage Alcohol monomester succinate/lipoic acid polymer micelle carrier (GCT).
Embodiment 3：The system of lentinan-cholesteryl hemisuccinate/oligomerization methionine polymer micelle carrier (LCM) It is standby
1.000g lentinans, 0.090g cholesteryl hemisuccinates are dissolved in formamide and N, N- dimethyl formyls respectively In amine mixed solvent (v/v=1: 1), under condition of ice bath, 0.077g EDC, 0.052g NHS, 0.009g DMAP activation courages are added 0.5~1h of free carboxy of sterol monomester succinate, is slowly dropped into the solution of lentinan, normal-temperature reaction 24h.Use excess Cold acetone precipitation, filter and purify sediment, redissolved with water, distilled water dialysis (MWCO=3500) 48~72h.Freeze-drying Obtain intermediate 1.
By 0.120g intermediates 1,0.200g oligomerizations methionine is dissolved in formamide respectively and n,N-Dimethylformamide is mixed In bonding solvent (v/v=1: 1), under condition of ice bath, 0.090g EDC, 0.060g NHS, 0.012g DMAP activation oligomerization first are added 0.5~1h of free carboxy of methyllanthionine, is slowly dropped into the solution of intermediate 1, normal-temperature reaction 24h.Use excessive cold acetone Precipitation, filters and purifies sediment, redissolved with water, distilled water dialysis (MWCO=3500) 48~72h.Freeze-drying obtains perfume (or spice) Mushroom polysaccharide-cholesteryl hemisuccinate/oligomerization methionine polymer micelle carrier (LCM).
Embodiment 4：The system of polyimides-cholesteryl hemisuccinate/oligomerization methionine polymer micelle carrier (PCM) It is standby
1.000g polyimides, 0.070g cholesteryl hemisuccinates are dissolved in n,N-Dimethylformamide respectively, ice Under the conditions of bath, 0.060g EDC, 0.5~1h of free carboxy of 0.040g NHS activation cholesterol monomester succinates are added, slowly Instill in the solution of polyimides, normal-temperature reaction 24h.Distilled water dialysis (MWCO=3500) 48~72h.Freeze-drying obtains Intermediate 1.
0.300g intermediates 1,0.320g oligomerizations methionine are dissolved in n,N-Dimethylformamide respectively, condition of ice bath Under, 0.5~1h of free carboxy of 0.144g EDC, 0.096g NHS activation oligomerization methionine is added, is slowly dropped into intermediate 1 Solution in, normal-temperature reaction 24h.Distilled water dialysis (MWCO=3500) 48~72h.Freeze-drying obtains polyimides-courage Sterol monomester succinate/oligomerization methionine polymer micelle carrier (PCM).
Embodiment 5：The preparation of glucan-cholesteryl hemisuccinate/poly- L-Histidine polymer micelle carrier (DCP)
1.000g glucans, 0.080g cholesteryl hemisuccinates are dissolved in formamide and n,N-Dimethylformamide respectively In mixed solvent (v/v=1: 1), under condition of ice bath, add 0.069g EDC, 0.046g NHS, 0.008g DMAP activation courages and consolidate 0.5~1h of free carboxy of alcohol monomester succinate, is slowly dropped into the solution of glucan, normal-temperature reaction 24h.Use the cold of excess Acetone precipitation, filters and purifies sediment, redissolved with water, distilled water dialysis (MWCO=3500) 48~72h.It is freeze-dried to obtain the final product To intermediate 1.
0.300g intermediates 1, the poly- L-Histidines of 0.100g are dissolved in dimethyl sulfoxide (DMSO) respectively, addition 0.055g EDC, 0.035g NHS, 0.004g DMAP activate 0.5~1h of free carboxy of poly- L-Histidine, are slowly dropped into the solution of intermediate 1 In, normal-temperature reaction 24h.Distilled water dialysis (MWCO=3500) 48~72h.Obtain glucan-cholesteryl hemisuccinate/poly- L-Histidine polymer micelle carrier (DCP).
Embodiment 6：The system of polyacrylic acid-cholesterol/4- hydroxy benzenes pinacol borate polymer micelle carriers (PACP) It is standby
1.000g polyacrylic acid, 0.090g cholesterol are dissolved in n,N-Dimethylformamide respectively, under condition of ice bath, added Enter 0.077g EDC, 0.052g NHS, 0.5~1h of free carboxy of 0.009g DMAP activated polyacrylic acids, be slowly dropped into courage and consolidate In the solution of alcohol, normal-temperature reaction 24h.Distilled water dialysis (MWCO=3500) 48~72h.Freeze-drying obtains intermediate 1.
0.120g intermediates 1,0.220g 4- hydroxy benzenes pinacol borates are dissolved in n,N-Dimethylformamide respectively In, under condition of ice bath, add 0.080g EDC, 0.050g NHS, 0.010g DMAP activated intermediates 1 free carboxy 0.5~ 1h, is slowly dropped into the solution of 4- hydroxy benzenes pinacol borates, normal-temperature reaction 24h.Distilled water dialysis (MWCO=3500) 48 ~72h.Freeze-drying obtains polyacrylic acid-cholesterol/4- hydroxy benzenes pinacol borate polymer micelle carriers (PACP).
Embodiment 7：The system of polymaleic anhydride-cholesterol/4- hydroxy benzenes pinacol borate polymer micelle carriers (PCP) It is standby
1.000g polymaleic anhydride, 0.080g cholesterol are dissolved in n,N-Dimethylformamide respectively, normal-temperature reaction 24h.Distilled water dialysis (MWCO=3500) 48~72h.Freeze-drying obtains intermediate 1.
0.300g intermediates 1,0.220g 4- hydroxy benzenes pinacol borates are dissolved in n,N-Dimethylformamide respectively In, normal-temperature reaction 24h.Distilled water dialysis (MWCO=3500) 48~72h.Freeze-drying i.e. obtain polymaleic anhydride-cholesterol/ 4- hydroxy benzenes pinacol borate polymer micelle carriers (PCP).
Embodiment 8：The preparation of hyaluronic acid-cholesterol/4- hydroxy benzenes pinacol borate polymer micelle carriers (HCP)
1.000g hyaluronic acids, 0.080g cholesterol are dissolved in formamide and n,N-Dimethylformamide mixed solvent respectively In (v/v=1: 1).Under condition of ice bath, the trip of 0.069g EDC, 0.046g NHS, 0.008g DMAP activation hyaluronic acids are added From 0.5~1h of carboxyl, it is slowly dropped into the solution of cholesterol, normal-temperature reaction 24h.Using excessive ice acetone precipitation, filter simultaneously Sediment is purified, is redissolved with water, distilled water dialysis (MWCO=3500) 48~72h.Freeze-drying obtains intermediate 1.
0.220g intermediates 1,0.220g 4- hydroxy benzenes pinacol borates are dissolved in formamide and N, N- dimethyl respectively In formamide mixed solvent (v/v=1: 1), under condition of ice bath, add 0.080g EDC, 0.050g NHS, 0.010g DMAP and live Change 0.5~1h of free carboxy of intermediate 1, be slowly dropped into the solution of 4- hydroxy benzenes pinacol borates, normal-temperature reaction 24h. Distilled water dialysis (MWCO=3500) 48~72h.Where freeze-drying obtains hyaluronic acid-cholesterol/4- hydroxyls phenyl boric acid frequency Ester polymer micellar carrier (HCP).
Embodiment 9：Hyaluronic acid-cholesteryl hemisuccinate/4,4 '-dicarboxyl azobenzene polymer micellar carrier (HCB) preparation
1.000g hyaluronic acids, 0.080g cholesteryl hemisuccinates are dissolved in formamide and N, N- dimethyl formyls respectively In amine mixed solvent (v/v=1: 1), under condition of ice bath, 0.069g EDC, 0.046g NHS, 0.008g DMAP activation courages are added After 0.5~1h of free carboxy of sterol monomester succinate, it is slowly dropped into the solution of hyaluronic acid, normal-temperature reaction 24h.Used The cold acetone precipitation of amount, filters and purifies sediment, redissolved with water, distilled water dialysis (MWCO=3500) 48~72h.Freezing is dry It is dry to obtain intermediate 1.
By 0.300g intermediates 1,0.320g 4,4 '-dicarboxyl azobenzene is dissolved in formamide and N, N- dimethyl methyls respectively In acid amides mixed solvent (v/v=1: 1), under condition of ice bath, 0.120g EDC, 0.080g NHS, 0.015g DMAP activation are added 4,0.5~1h of free carboxy of 4 '-dicarboxyl azobenzene, is slowly dropped into the solution of intermediate 1, normal-temperature reaction 24h.Used The cold acetone precipitation of amount, filters and purifies sediment, redissolved with water, distilled water dialysis (MWCO=3500) 48~72h.Freezing is dry It is dry to obtain hyaluronic acid-cholesteryl hemisuccinate/4,4 '-dicarboxyl azobenzene polymer micellar carrier (HCB).
Embodiment 10：Chitosan-cholesteryl hemisuccinate/4,4 '-dicarboxyl azobenzene polymer micellar carrier (GCB) Preparation
1.000g chitosans, 0.080g cholesteryl hemisuccinates are dissolved in dimethyl sulfoxide (DMSO) respectively, under condition of ice bath, 0.069g EDC, 0.046g NHS, 0.5~1h of free carboxy of 0.008g DMAP activation cholesterol monomester succinates are added, is delayed It is slow to instill in the solution of chitosan, normal-temperature reaction 24h.Using excessive cold acetone precipitation, filter and purify sediment, answered with water It is molten, distilled water dialysis (MWCO=3500) 48~72h.Freeze-drying obtains intermediate 1.
By 0.300g intermediates 1,0.320g 4,4 '-dicarboxyl azobenzene is dissolved in formamide and N, N- dimethyl methyls respectively In acid amides mixed solvent (v/v=1: 1), under condition of ice bath, 0.120g EDC, 0.080g NHS, 0.015g DMAP activation are added 4,0.5~1h of free carboxy of 4 '-dicarboxyl azobenzene, is slowly dropped into the solution of intermediate 1, normal-temperature reaction 24h.Used The cold acetone precipitation of amount, filters and purifies sediment, redissolved with water, distilled water dialysis (MWCO=3500) 48~72h.Freezing is dry It is dry to obtain chitosan-cholesteryl hemisuccinate/4,4 '-dicarboxyl azobenzene polymer micellar carrier (GCB).
Embodiment 11：The preparation of the different efficient cross-film polymer micelle of molecular modification type and characterization
1. the preparation of the different efficient cross-film polymer micelle of molecular modification type：The different efficient cross-film polymer micelle of molecular modification type Carrier lyophilized products 30mg, which is dissolved in 6mL pure water, is stirred at room temperature 1h, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of filter membrane mistake Filter, to obtain the final product.
2. the measure of self-assembled micelle critical micelle concentration (CMC)：The critical micelle concentration (CMC) of micella refers to amphipathic The least concentration that aggregation is self-assembly of micella occurs in a solvent for molecule.Fluorescence probe method accuracy height and high sensitivity, lead to Frequently with the CMC value of this method measure micella.Pyrene is a kind of hydrophobic condensed-nuclei aromatics class compound, and solubility in water is very Difference, and micella cannot be formed in the solution, pyrene micro at this time is dissolved in the water system of polarity；When the concentration of amphipathic molecule is high During CMC, you can form micellar structure, at this time, hydrophobic pyrene is distributed as the hydrophobic region of micelle inner core, and enters nonpolar ring A series of changes are presented in border, its fluorescence spectrum, such as：Fluorescence intensity increases, and the vibration fine structure in emission spectrum changes, (0,0) wave band red shift in excitation spectrum, 338nm is moved to from 333nm.Therefore, I in the excitation spectrum with pyrene is passed through338/I333Ratio (excitation spectrum medium wavelength is respectively the corresponding fluorescence intensity levels of 338nm and 333nm) maps the log concentration of amphiphilic species The CMC of amphiphilic species is can obtain, the results are shown in Table 1.
The measure of the critical micelle concentration (CMC) of the efficient cross-film polymer micelle of the different molecular modification type of table 1
3. particle diameter：3000 HS instrument of Zetasizer (Malvern Instruments, Malvern, UK) exist 633nm/25 DEG C, He-Ne laser determination sample particle diameters, the results are shown in Table 2.
The diameter characterization of the efficient cross-film polymer micelle of the different molecular modification type of table 2
Embodiment 12：Contain hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle of taxol (HCT/PTX) preparation and characterization
1. preparation process
(1) Probe Ultrasonic Searching method
By hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (HCT) lyophilized products 30mg dissolvings It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Taxol 20mg is dissolved in nothing In water-ethanol.The two solution is mixed, it is saturating with bag filter (MWCO=3500) room temperature in distilled water after ice-bath ultrasonic 30min 12h is analysed, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
(2) high pressure homogenization method
Hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (HCT) lyophilized products 30mg is dissolved in It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Taxol 20mg is dissolved in nothing In water-ethanol.The two solution is mixed, after high-pressure homogeneous, with bag filter (MWCO=3500), room temperature is dialysed 12h in distilled water, 5~10min of 3000rpm are centrifuged, with 0.8 μm of membrane filtration, freeze-drying.
(3) solvent evaporation method
Hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (HCT) lyophilized products 30mg is dissolved in It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Taxol 20mg is dissolved in nothing In water-ethanol.The two solution is mixed, is stirred overnight, absolute ethyl alcohol is volatilized, 5~10min of 3000rpm are centrifuged, with 0.8 μm Membrane filtration, freeze-drying.
2. hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle (HCT/PTX) contains taxol and contains The measure of amount.
Using the content of high performance liquid chromatography-uv detection method measure taxol.Chromatographic column is Lichrospher C18 (4.6mm*250mm, 5 μm), mobile phase are methanol-water (70: 30, v/v), and flow velocity 1mL/min, column temperature is 30 DEG C, detects ripple A length of 227nm, sample size are 20 μ l.Drugloading rate (the EL of taxol in sample is calculated with formula (1)PTX(%)).
3. particle diameter：3000 HS instrument of Zetasizer (Malvern Instruments, Malvern, UK) exist 633nm/25 DEG C, He-Ne laser determination sample particle diameters.
Embodiment 12 (1)~12 (3) contains hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymer of taxol Carrier micelle (HCT/PTX) physicochemical property is shown in Table 3.
Table 3 contains hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle (HCT/ of taxol PTX characterization)
Embodiment 13：Contain chitosan-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle of taxol (GCT/PTX) preparation and characterization
1. preparation process
(1) Probe Ultrasonic Searching method
Chitosan-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (GCT) lyophilized products 30mg is dissolved in It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Taxol 20mg is dissolved in nothing In water-ethanol.The two solution is mixed, it is saturating with bag filter (MWCO=3500) room temperature in distilled water after ice-bath ultrasonic 30min 12h is analysed, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
(2) high pressure homogenization method
Chitosan-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (GCT) lyophilized products 30mg is dissolved in It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Taxol 20mg is dissolved in nothing In water-ethanol.The two solution is mixed, after high-pressure homogeneous, with bag filter (MWCO=3500), room temperature is dialysed 12h in distilled water, 5~10min of 3000rpm are centrifuged, with 0.8 μm of membrane filtration, freeze-drying.
(3) solvent evaporation method
Chitosan-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (GCT) lyophilized products 30mg is dissolved in It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Taxol 20mg is dissolved in nothing In water-ethanol.The two solution is mixed, is stirred overnight, absolute ethyl alcohol is volatilized, 5~10min of 3000rpm are centrifuged, with 0.8 μm Membrane filtration, freeze-drying.
2. chitosan-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle (GCT/PTX) contains content of taxol Measure.
Using the content of high performance liquid chromatography-uv detection method measure taxol.Chromatographic column is Lichrospher C18 (4.6mm*250mm, 5 μm), mobile phase are methanol-water (70: 30, v/v), and flow velocity 1mL/min, column temperature is 30 DEG C, detects ripple A length of 227nm, sample size are 20 μ l.Drugloading rate (the EL of taxol in sample is calculated with formula (1)PTX(%)).
3. particle diameter：3000 HS instrument of Zetasizer (Malvern Instruments, Malvern, UK) exist 633nm/25 DEG C, He-Ne laser determination sample particle diameters.
Chitosan-cholesteryl hemisuccinate/lipoic acid that embodiment 13 (1)~13 (3) contains taxol is polymer supported Medicine micella (GCT/PTX) physicochemical property is shown in Table 4.
Table 4 contains chitosan-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle (GCT/PTX) of taxol Characterization
Embodiment 14：Contain hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle of legalon (HCT/SB) preparation and characterization
1. preparation process
(1) Probe Ultrasonic Searching method
Hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (HCT) lyophilized products 30mg is dissolved in It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Legalon 20mg is dissolved in In acetone.The two solution is mixed, after ice-bath ultrasonic 30min, with bag filter (MWCO=3500), room temperature is dialysed in distilled water 12h, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
(2) high pressure homogenization method
Hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (HCT) lyophilized products 30mg is dissolved in It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Legalon 20mg is dissolved in In acetone.The two solution is mixed, after high-pressure homogeneous, with bag filter (MWCO=3500), room temperature is dialysed 12h in distilled water, from The heart 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
(3) solvent evaporation method
Hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (HCT) lyophilized products 30mg is dissolved in It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Legalon 20mg is dissolved in In acetone.The two solution is mixed, is stirred overnight, acetone is volatilized, 5~10min of 3000rpm are centrifuged, with 0.8 μm of filter membrane mistake Filter, freeze-drying.
2. hyaluronic acid-cholesteryl hemisuccinate/lipoic acid carrier micelle (HCT/SB) contains legalon content Measure.
The content of assay legalon is carried out using high performance liquid chromatography-uv detection method.Chromatographic column is Lichrospher C18(4.6mm*250mm, 5 μm), mobile phase are methanol-water-glacial acetic acid (40: 60: 1, v/v), and flow velocity is 1mL/min, column temperature are 30 DEG C, Detection wavelength 288nm, and sample size is 20 μ l.Legalon in sample is calculated with formula (1) Drugloading rate (ELSB(%)).
3. particle diameter：3000 HS instrument of Zetasizer (Malvern Instruments, Malvern, UK) exist 633nm/25 DEG C, He-Ne laser determination sample particle diameters.
Embodiment 14 (1)~14 (3) is loaded with hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymerization of legalon Thing carrier micelle (HCT/SB) physicochemical property is shown in Table 5.
Table 5 is loaded with hyaluronic acid-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle (HCT/ of legalon SB characterization)
Embodiment 15：Contain chitosan-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle of legalon (GCT/SB) preparation and characterization
1. preparation process
(1) Probe Ultrasonic Searching method
Chitosan-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (GCT) lyophilized products 30mg is dissolved in It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Legalon 20mg is dissolved in In acetone.The two solution is mixed, after ice-bath ultrasonic 30min, with bag filter (MWCO=3500), room temperature is dialysed in distilled water 12h, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
(2) high pressure homogenization method
Chitosan-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (GCT) lyophilized products 30mg is dissolved in It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Legalon 20mg is dissolved in In acetone.The two solution is mixed, after high-pressure homogeneous, with bag filter (MWCO=3500), room temperature is dialysed 12h in distilled water, from The heart 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
(3) solvent evaporation method
Chitosan-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (GCT) lyophilized products 30mg is dissolved in It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Legalon 20mg is dissolved in In acetone.The two solution is mixed, is stirred overnight, acetone is volatilized, 5~10min of 3000rpm are centrifuged, with 0.8 μm of filter membrane mistake Filter, freeze-drying.
2. chitosan-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle (GCT/SB) contains legalon and contains The measure of amount.
The content of assay legalon is carried out using high performance liquid chromatography-uv detection method.Chromatographic column is Lichrospher C18 (4.6mm*250mm, 5 μm), mobile phase are methanol-water-glacial acetic acid (40: 60: 1, v/v), and flow velocity is 1mL/min, column temperature are 30 DEG C, Detection wavelength 288nm, and sample size is 20 μ l.Legalon in sample is calculated with formula (1) Drugloading rate (ELSB(%)).
3. particle diameter：3000 HS instrument of Zetasizer (Malvern Instruments, Malvern, UK) exist 633nm/25 DEG C, He-Ne laser determination sample particle diameters.
Embodiment 15 (1)~15 (3) is loaded with chitosan-cholesteryl hemisuccinate/lipoic acid polymer of legalon The physicochemical property of carrier micelle (GCT/SB) is shown in Table 6.
Table 6 is loaded with chitosan-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle (GCT/SB) of legalon Characterization
Embodiment 16：Contain polymaleic anhydride-cholesterol/4- hydroxy benzenes pinacol borate carrier micelles of astaxanthin (PCP/ASTA) preparation and characterization
1. preparation process
(1) Probe Ultrasonic Searching method
By polymaleic anhydride-cholesterol/4- hydroxy benzenes pinacol borate polymer micelle carrier (PCP) lyophilized products 30mg It is dissolved in 6mL pure water and 1h is stirred at room temperature, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Astaxanthin 20mg is molten Solution is in methyl alcohol.The two solution is mixed, after ice-bath ultrasonic 30min, with bag filter (MWCO=3500) room temperature in distilled water Dialyse 12h, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
(2) high pressure homogenization method
By polymaleic anhydride-cholesterol/4- hydroxy benzenes pinacol borate polymer micelle carrier (PCP) lyophilized products 30mg It is dissolved in 6mL pure water and 1h is stirred at room temperature, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Astaxanthin 20mg is molten Solution is in methyl alcohol.The two solution is mixed, after high-pressure homogeneous, with bag filter (MWCO=3500), room temperature is dialysed in distilled water 12h, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
(3) solvent evaporation method
By polymaleic anhydride-cholesterol/4- hydroxy benzenes pinacol borate polymer micelle carrier (PCP) lyophilized products 30mg It is dissolved in 6mL pure water and 1h is stirred at room temperature, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Astaxanthin 20mg is molten Solution is in methyl alcohol.The two solution is mixed, is stirred overnight, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freezing It is dry.
2. polymaleic anhydride-cholesterol/4- hydroxy benzenes pinacol borate polymer medicament carrying micelles (PCP/ASTA) contain The measure of content astaxanthin.
The content of assay astaxanthin is carried out using high performance liquid chromatography-uv detection method.Chromatographic column is Lichrospher C18(4.6mm*250mm, 5 μm), mobile phase are methanol-water (95: 5, v/v), flow velocity 1mL/min, column temperature For 30 DEG C, Detection wavelength 475nm, sample size is 20 μ l.Drugloading rate (the EL of astaxanthin in sample is calculated with formula (1)ASTA (%)).
3. particle diameter：3000 HS instrument of Zetasizer (Malvern Instruments, Malvern, UK) exist 633nm/25 DEG C, He-Ne laser determination sample particle diameters.
Embodiment 16 (1)~16 (3) is loaded with polymaleic anhydride-cholesterol/4- hydroxy benzenes pinacol borates of astaxanthin Polymer medicament carrying micelle (PCP/ASTA) physicochemical property is shown in Table 7.
Table 7 is loaded with polymaleic anhydride-cholesterol/4- hydroxy benzenes pinacol borate polymer medicament carrying micelles of astaxanthin (PCP/ASTA) characterization
Embodiment 17：Contain polyacrylic acid-cholesterol/polymer drug-carried glue of 4- hydroxy benzenes pinacol borates of erythromycin The preparation of beam (PACP/ERY) and characterization
1. preparation process
(1) Probe Ultrasonic Searching method
By polyacrylic acid-cholesterol/4- hydroxy benzenes pinacol borate polymer micelle carrier (PACP) lyophilized products 30mg It is dissolved in 6mL pure water and 1h is stirred at room temperature, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Erythromycin 20mg is molten Solution is in n,N-Dimethylformamide, and after ice-bath ultrasonic 30min, with bag filter (MWCO=3500), room temperature is dialysed in distilled water 12h, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
(2) high pressure homogenization method
By polyacrylic acid-cholesterol/4- hydroxy benzenes pinacol borate polymer micelle carrier (PACP) lyophilized products 30mg It is dissolved in 6mL pure water and 1h is stirred at room temperature, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Erythromycin 20mg is molten Solution is in n,N-Dimethylformamide.The two solution is mixed, after high-pressure homogeneous, with bag filter (MWCO=3500) in distilled water Middle room temperature dialysis 12h, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
2. polyacrylic acid-cholesterol/4- hydroxy benzenes pinacol borate polymer medicament carrying micelles (PACP/ERY) contain red The measure of mycin content.
Assay, chromatographic column Lichrospher are carried out to erythromycin using high performance liquid chromatography-uv detection method C18(4.6mm*250mm, 5 μm), (is taken dipotassium hydrogen phosphate 8.7g, adds water 1000mL, with phosphorus acid for adjusting pH value extremely with phosphate solution 8.2)-acetonitrile (40: 60) is mobile phase；Flow velocity is 1mL/min, 35 DEG C of column temperature；Wavelength is 215nm, and sample size is 20 μ l.With public affairs Formula (1) calculates the drugloading rate (EL of erythromycin in sampleDOX(%)).
3. particle diameter：3000 HS instrument of Zetasizer (Malvern Instruments, Malvern, UK) exist 633nm/25 DEG C, He-Ne laser determination sample particle diameters.
Polyacrylic acid-cholesterol/4- hydroxy benzenes pinacol borates that embodiment 17 (1)~17 (2) is loaded with erythromycin gather Compound carrier micelle (PACP/ERY) physicochemical property is shown in Table 8.
Table 8 is loaded with polyacrylic acid-cholesterol/4- hydroxy benzenes pinacol borate polymer medicament carrying micelles of erythromycin (PACP/ERY) characterization
Embodiment 18：Polyimides-cholesteryl hemisuccinate/oligomerization the methionine for containing Allopurinol is polymer drug-carried The preparation of micella (PCM/ALO) and characterization
1. preparation process
(1) Probe Ultrasonic Searching method
By polyimides-cholesteryl hemisuccinate/oligomerization methionine polymer micelle carrier (PCM) lyophilized products 30mg It is dissolved in 6mL pure water and 1h is stirred at room temperature, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Allopurinol 20mg is molten Solution is in methyl alcohol.The two solution is mixed, after ice-bath ultrasonic 30min, with bag filter (MWCO=3500) room temperature in distilled water Dialyse 12h, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
(2) high pressure homogenization method
By polyimides-cholesteryl hemisuccinate/oligomerization methionine polymer micelle carrier (PCM) lyophilized products 30mg It is dissolved in 6mL pure water and 1h is stirred at room temperature, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Allopurinol 20mg is molten Solution is in methyl alcohol.The two solution is mixed, after high-pressure homogeneous, with bag filter (MWCO=3500), room temperature is dialysed in distilled water 12h, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
(3) solvent evaporation method
By polyimides-cholesteryl hemisuccinate/oligomerization methionine polymer micelle carrier (PCM) lyophilized products 30mg It is dissolved in 6mL pure water and 1h is stirred at room temperature, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Allopurinol 20mg is molten Solution is in methyl alcohol.The two solution is mixed, is stirred overnight, methanol is volatilized, centrifuges 5~10min of 3000rpm, with 0.8 μm of filter Membrane filtration, freeze-drying.
2. polyimides-cholesteryl hemisuccinate/oligomerization methionine polymer medicament carrying micelle (PCM/ALO) contains not The measure of fast alcohol content.
The content of assay Allopurinol is carried out using high performance liquid chromatography-uv detection method.Chromatographic column is Lichrospher C18(4.6mm*250mm, 5 μm), mobile phase are methanol-water-glacial acetic acid (75: 25: 0.2), flow velocity 1mL/ Min, column temperature are 30 DEG C, Detection wavelength 251nm, and sample size is 20 μ l.The drugloading rate of Allopurinol in sample is calculated with formula (1) (ELALO (%)).
3. particle diameter：3000 HS instrument of Zetasizer (Malvern Instruments, Malvern, UK) exist 633nm/25 DEG C, He-Ne laser determination sample particle diameters.
Embodiment 18 (1)~18 (3) is loaded with polyimides-cholesteryl hemisuccinate/oligomerization methionine of Allopurinol Polymer medicament carrying micelle (PCM/ALO) physicochemical property is shown in Table 9.
Table 9 is loaded with polyimides-cholesteryl hemisuccinate/oligomerization methionine polymer medicament carrying micelle of Allopurinol (PCM/ALO) characterization
Embodiment 19：Glucan-cholesteryl hemisuccinate/poly- L-Histidine of taxol and legalon is contained at the same time The preparation of polymer medicament carrying micelle (DCP/PTX+SB) and characterization
1. preparation process
(1) Probe Ultrasonic Searching method
By glucan-cholesteryl hemisuccinate/poly- L-Histidine polymer micelle carrier (DCP) lyophilized products 30mg dissolvings It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Taxol and legalon Each 20mg dissolves in acetone at the same time.The two solution is mixed, after ice-bath ultrasonic 30min, is existed with bag filter (MWCO=3500) Room temperature dialysis 12h, centrifuges 5~10min of 3000rpm in distilled water, with 0.8 μm of membrane filtration, freeze-drying.
(2) high pressure homogenization method
By glucan-cholesteryl hemisuccinate/poly- L-Histidine polymer micelle carrier (DCP) lyophilized products 30mg dissolvings It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Taxol and legalon Each 20mg dissolves in acetone at the same time.The two solution is mixed, after high-pressure homogeneous, with bag filter (MWCO=3500) in distilled water Middle room temperature dialysis 12h, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
(3) solvent evaporation method
By glucan-cholesteryl hemisuccinate/poly- L-Histidine polymer micelle carrier (DCP) lyophilized products 30mg dissolvings It is stirred at room temperature 1h in 6mL pure water, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Taxol and legalon Each 20mg dissolves in acetone at the same time.The two solution is mixed, is stirred overnight, methanol is volatilized, centrifugation 3000rpm 5~ 10min, with 0.8 μm of membrane filtration, freeze-drying.
2. glucan-cholesteryl hemisuccinate/poly- L-Histidine polymer medicament carrying micelle (DCP/PTX+SB) wraps at the same time Carry the measure of taxol and legalon content.
Assay is carried out to taxol and legalon using high performance liquid chromatography-uv detection method respectively.Chromatography Column is Lichrospher C18(4.6mm*250mm, 5 μm), mobile phase are methanol-water (70: 30, v/v), flow velocity 1mL/min, Column temperature is 30 DEG C, Detection wavelength 227nm, and sample size is 20 μ l.The drugloading rate of taxol in sample is calculated with formula (1) (ELPTX(%))；Chromatographic column is Lichrospher C18(4.6mm*250mm, 5 μm), mobile phase for methanol-water-glacial acetic acid (40: 60: 1, v/v), flow velocity 1mL/min, column temperature is 30 DEG C, Detection wavelength 288nm, and sample size is 20 μ l..In terms of formula (1) Calculate the drugloading rate (EL of legalon in sampleSB(%)).
3. particle diameter：3000 HS instrument of Zetasizer (Malvern Instruments, Malvern, UK) exist 633nm/25 DEG C, He-Ne laser determination sample particle diameters.
Embodiment 19 (1)~19 (3) at the same contain glucan-cholesteryl hemisuccinate of taxol and legalon/ Poly- L-Histidine polymer medicament carrying micelle (DCP/PTX+SB) physicochemical property is shown in Table 10.
Table 10 contains glucan-cholesteryl hemisuccinate/poly- L-Histidine polymerization of taxol and legalon at the same time The characterization of thing carrier micelle (DCP/PTX+SB)
Embodiment 20：Polyimides-cholesteryl hemisuccinate/oligomerization first sulphur of adriamycin and legalon is contained at the same time The preparation of propylhomoserin polymer medicament carrying micelle (PCM/DOX+SB) and characterization
1. preparation process
(1) Probe Ultrasonic Searching method
By polyimides-cholesteryl hemisuccinate/oligomerization methionine polymer micelle carrier (PCM) lyophilized products 30mg It is dissolved in 6mL pure water and 1h is stirred at room temperature, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Desalination adriamycin and Each 20mg of legalon is dissolved in acetone and n,N-Dimethylformamide mixed solvent (v/v=1: 1) at the same time, ice-bath ultrasonic After 30min, with bag filter (MWCO=3500) in distilled water room temperature dialyse 12h, centrifuge 5~10min of 3000rpm, with 0.8 μ M membrane filtrations, freeze-drying.
(2) high pressure homogenization method
By polyimides-cholesteryl hemisuccinate/oligomerization methionine polymer micelle carrier (PCM) lyophilized products 30mg It is dissolved in 6mL pure water and 1h is stirred at room temperature, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Desalination adriamycin and Each 20mg of legalon is dissolved in acetone and n,N-Dimethylformamide mixed solvent (v/v=1: 1) at the same time, will the two solution Mixing, after high-pressure homogeneous, with bag filter (MWCO=3500) in distilled water room temperature dialyse 12h, centrifugation 3000rpm 5~ 10min, with 0.8 μm of membrane filtration, freeze-drying.
2. polyimides-cholesteryl hemisuccinate/oligomerization methionine polymer medicament carrying micelle micellar carrier (PCM/ DOX+SB) while the measure of adriamycin and legalon content is contained.
Carrying out the content of assay adriamycin using fluophotometer, (excitation wavelength and launch wavelength are respectively 488nm And 570nm).Drugloading rate (the EL of adriamycin in sample is calculated with formula (1)DOX(%)).Using high performance liquid chromatography-ultraviolet Detection method carries out assay to legalon.Chromatographic column is Lichrospher C18(4.6mm*250mm, 5 μm), mobile phase is Methanol-water-glacial acetic acid (40: 60: 1, v/v), flow velocity 1mL/min, column temperature are 30 DEG C, Detection wavelength 288nm, and sample size is 20μl.Drugloading rate (the EL of legalon in sample is calculated with formula (1)SB(%)).
3. particle diameter：3000 HS instrument of Zetasizer (Malvern Instruments, Malvern, UK) exist 633nm/25 DEG C, He-Ne laser determination sample particle diameters.
Embodiment 20 (1)~20 (2) at the same be loaded with adriamycin and legalon polyimides-cholesterol succinic acid it is single Ester/oligomerization methionine polymer medicament carrying micelle (PCM/DOX+SB) physicochemical property is shown in Table 11.
Table 11 is loaded with polyimides-cholesteryl hemisuccinate/oligomerization methionine of adriamycin and legalon at the same time The characterization of polymer medicament carrying micelle (PCM/DOX+SB)
Embodiment 21：Chitosan-cholesteryl hemisuccinate/4 of adriamycin and taxol, 4 '-dicarboxyl are contained at the same time The preparation of azobenzene polymer carrier micelle (GCB/DOX+PTX) and characterization
1. preparation process
(1) Probe Ultrasonic Searching method
By chitosan-cholesteryl hemisuccinate/4,4 '-dicarboxyl azobenzene polymer micellar carrier (GCB) lyophilized products 30mg, which is dissolved in 6mL pure water, is stirred at room temperature 1h, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Desalination Ah mould Element and each 20mg of taxol are dissolved in n,N-Dimethylformamide at the same time, after ice-bath ultrasonic 30min, with bag filter (MWCO= 3500) the room temperature dialysis 12h in distilled water, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freeze-drying.
(2) high pressure homogenization method
By chitosan-cholesteryl hemisuccinate/4,4 '-dicarboxyl azobenzene polymer micellar carrier (GCB) lyophilized products 30mg, which is dissolved in 6mL pure water, is stirred at room temperature 1h, after ultrasonic or high-pressure homogeneous under ice bath, 0.8 μm of membrane filtration.Desalination Ah mould Element and each 20mg of taxol are dissolved in n,N-Dimethylformamide at the same time.The two solution is mixed, after high-pressure homogeneous, with dialysis Bag (MWCO=3500) room temperature dialysis 12h in distilled water, centrifuges 5~10min of 3000rpm, with 0.8 μm of membrane filtration, freezing It is dry.
2. chitosan-cholesteryl hemisuccinate/4,4 '-dicarboxyl azobenzene polymer carrier micelle (GCB/DOX+ PTX) while the measure of adriamycin and legalon content is contained.
Carrying out the content of assay adriamycin using fluophotometer, (excitation wavelength and launch wavelength are respectively 488nm And 570nm).Drugloading rate (the EL of adriamycin in sample is calculated with formula (1)DOX(%)).Using high performance liquid chromatography-ultraviolet Detection method carries out assay to taxol, and chromatographic column is Lichrospher C18(4.6mm*250mm, 5 μm), mobile phase is first Alcohol-water (70: 30, v/v), flow velocity 1mL/min, column temperature are 30 DEG C, Detection wavelength 227nm, and sample size is 20 μ l.With formula (1) drugloading rate (EL of taxol in sample is calculatedPTX(%)).
3. particle diameter：3000 HS instrument of Zetasizer (Malvern Instruments, Malvern, UK) exist 633nm/25 DEG C, He-Ne laser determination sample particle diameters.
Embodiment 21 (1)~21 (2) while chitosan-cholesteryl hemisuccinate/4 of adriamycin and taxol are loaded with, 4 '-dicarboxyl azobenzene polymer carrier micelle (GCB/DOX+PTX) physicochemical property is shown in Table 12.
Table 12 is loaded with chitosan-cholesteryl hemisuccinate/4 of adriamycin and taxol, 4 '-dicarboxyl azobenzene at the same time The characterization of polymer medicament carrying micelle (GCB/DOX+PTX)
Embodiment 22：Contain chitosan-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle of taxol (GCT/PTX) and contain taxol chitosan-lipoic acid polymer medicament carrying micelle (GT/PTX) release in vitro
1. the preparation of polymer micelle carrier
(1) preparation of chitosan-cholesteryl hemisuccinate/lipoic acid polymer micelle carrier (GCT)
Preparation method according to above-described embodiment 2 prepares chitosan-cholesteryl hemisuccinate/lipoic acid polymer micelle Carrier (GCT).
(2) preparation of chitosan-lipoic acid polymer micelle carrier (GT)
1.000g chitosans, 0.220g lipoic acids are dissolved in dimethyl sulfoxide (DMSO), add 0.990g EDC, 0.589g NHS, 0.015g DMAP activate 0.5~1h of free carboxy of lipoic acid, are slowly dropped into the solution of chitosan, normal-temperature reaction 24h.Distilled water dialysis (MWCO=3500) 48~72h.Freeze-drying obtains chitosan-lipoic acid polymer micelle carrier (GT)。
2. contain the preparation of the polymer medicament carrying micelle of taxol
(1) chitosan-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle (GCT/PTX) of taxol is contained Preparation
Preparation method according to above-described embodiment 13 (1) prepare contain chitosan-cholesteryl hemisuccinate of taxol/ Lipoic acid polymer medicament carrying micelle (GCT/PTX).
(2) preparation of chitosan-lipoic acid polymer medicament carrying micelle (GT/PTX) of taxol is contained
Chitosan-lipoic acid polymer (GT) lyophilized products 30mg is dissolved in 6mL pure water and 1h is stirred at room temperature, under ice bath After ultrasonic or high-pressure homogeneous, 0.8 μm of membrane filtration.Taxol 20mg is dissolved in absolute ethyl alcohol.The two solution is mixed, ice bath After ultrasonic 30min, with bag filter (MWCO=3500) in distilled water room temperature dialyse 12h, centrifuge 5~10min of 3000rpm, use 0.8 μm of membrane filtration, is freeze-dried, and contains chitosan-lipoic acid polymer medicament carrying micelle (GT/PTX) of taxol to obtain the final product.
3. release in vitro
Chitosan-cholesteryl hemisuccinate/lipoic acid polymer medicament carrying micelle (the GCT/ for containing taxol is taken respectively PTX) and chitosan-lipoic acid polymer medicament carrying micelle (GT/PTX) of taxol is contained in right amount, it is dilute with 5% glucose solution PTX concentration is released to 0.5mg/mL.The phosphate-buffered containing 10 μM, the glutathione of 10mM and pH=7.4 is placed on respectively In the dissolution medium of liquid (Tween 80 for containing 0.2%).1mLGCT/PTX, GT/PTX polypeptide drug-loaded micelle solution is respectively taken to be placed in bag filter (MWCO=3500) in, it is put in the beaker for filling 200mL dissolution mediums.Water-bath constant temperature oscillator setting speed for 100 ± 5rpm, temperature carry out the extracorporeal releasing test of carrier micelle under the conditions of being 37 ± 0.5 DEG C.According to preset time point (0,1,2,3, 4th, 6,10,16,24,36,48h) take 1mL dissolution mediums to come out, and the fresh dissolution medium for supplementing same volume is entered.Ensure to burn Medium volume in cup is always 200mL.
The release sample of acquirement is stored and freezed in -20 DEG C, is redissolved when to be determined with proper amount of methanol, 0.22 μm of filter membrane mistake The content of assay taxol is directly carried out after filter using high performance liquid chromatography-uv detection method.Chromatographic column is Lichrospher C18(4.6mm*250mm, 5 μm), mobile phase are methanol-water (70: 30, v/v), flow velocity 1mL/min, column temperature For 30 DEG C, Detection wavelength 227nm, sample size is 20 μ l.The medicament contg of PTX in dissolution medium is measured, and is calculated respectively pre- If the accumulative release percentage of time point each medicine, release profiles, the result is shown in Figure 1 are drawn.Under the conditions of 10 μM of glutathione, GCT/PTX and GT/PTX are relatively stable, and burst size is smaller；And under the conditions of 10mM glutathione, GCT/PTX and GT/PTX's Under the conditions of obviously higher than 10 μM glutathione of rate of release and Cumulative release amount, show lesion microenvironment sensitivity disulfide bond herein Under the conditions of specificly-response degrade, the rate of release of GCT/PTX is more than GT/PTX in addition, shows the different efficient cross-film of molecular modification type Polymer medicament carrying micelle GCT/PTX is in lesion microenvironment, lesion microenvironment sensitivity key selective degradation, and the particle diameter of micella increases Greatly, the hydrophobic forces of kernel weaken, and realize medicine quick-release；And the polymer medicament carrying micelle GT/ of single sensitive hydrophobic molecule modification The nano-micelle that PTX responses are partially destroyed after stimulating exists in the form of precipitating, and drug release rate is not so good as target micella.

Claims (9)

1. a kind of efficient cross-film polymer micelle of different molecular modification type, it is characterised in that the micellar carrier is by hydrophily height The hydrophobic molecule for having efficient cross-film function is first introduced on molecule chain backbone, further introduces and contains to hydrophilic macromolecule chain backbone The hydrophobic molecule of lesion microenvironment sensitivity key builds to obtain；The carrier have it is amphipathic, can in an aqueous medium self assembly to receive Rice glue beam, can be wrapped in micelle inner core by hydrophobic drug by noncovalent interaction and deliver it to up to lesions position；Glue The hydrophobic molecule of the efficient cross-film function of tool in binding structure, can mediate the efficient cross-film of medicament-carried nano micelle to enter born of the same parents；In addition, micella knot Lesion microenvironment sensitivity key contained by structure can cause the hydrophobic work of medicament-carried nano micelle kernel by focal zone microenvironment selective degradation Firmly weaken, particle diameter increase, medicine, which is fast released, acts on focal zone, is remarkably improved the dense of lesions position free drug Degree, curative effect and bioavilability, the micellar carrier general structure are as follows：
[R1]q-M-[R2]p
Wherein M is hydrophilic macromolecule chain；R1To have the hydrophobic molecule of efficient cross-film function；R2For the sensitivity key of microenvironment containing lesion Hydrophobic molecule；Q is the substitution value for the hydrophobic molecule for having efficient cross-film function on hydrophilic macromolecule chain backbone, and the tool is efficient The substitution value of the hydrophobic molecule of cross-film function is 2%~25%；P is sensitive for microenvironment containing lesion on hydrophilic macromolecule chain backbone The substitution value of the hydrophobic molecule of key, the substitution value of the hydrophobic molecule of the sensitivity of microenvironment containing the lesion key is 2%~70%.
2. a kind of different efficient cross-film polymer micelle carrier of molecular modification type as claimed in claim 1, it is characterised in that hydrophilic Property macromolecular chain be selected from hyaluronic acid, unfraction heparin, low molecular weight heparin, desulfated heparin, chondroitin, poly-sulfated soft Ossein, alginic acid, glucan, fungi polysaccharide, chitosan, lentinan, polymaleic anhydride, polyacrylic acid and containing carboxyl, One kind in the derivative of amino or hydroxyl.
3. a kind of different efficient cross-film polymer micelle carrier of molecular modification type as claimed in claim 1, it is characterised in that pass through Amido link or ester bond have the hydrophobic molecule for having efficient cross-film function to be selected from cholesterol, cholesterol in hydrophilic macromolecule backbone modification One kind in derivative, wherein cholesterol derivative are selected from butanedioic acid cholesterol monoesters, 3 beta-amino -5- cholestene, O-phthalic One kind in sour cholesterol hydrogen ester.
4. a kind of different efficient cross-film polymer micelle carrier of molecular modification type as claimed in claim 1, it is characterised in that pass through Amido link or ester bond have the hydrophobic molecule of the sensitivity key of microenvironment containing lesion in hydrophilic macromolecule backbone modification, wherein micro- containing lesion The hydrophobic molecule of environment sensitive key be selected from lipoic acid, 4- hydroxy benzenes pinacol borate, 4,4 '-dicarboxyl azobenzene, azobenzene- One kind in 4 benzoic acid, poly- L-Histidine, oligomerization methionine.
5. a kind of preparation method of the different efficient cross-film polymer micelle carrier of molecular modification type as claimed in claim 1, it is special Sign is to comprise the following steps：
Hydrophilic macromolecule chain containing amino, carboxyl or hydroxyl is dissolved in reaction dissolvent, using activator will have efficiently across The hydrophobic molecule of film function obtains intermediate 1 by amido link or linkage in hydrophilic macromolecule；Intermediate 1 is dissolved in instead Answer in solvent, using activator by the hydrophobic molecule of the sensitivity key of microenvironment containing lesion by amido link or linkage in intermediate 1, that is, obtain the efficient cross-film polymer micelle carrier of different molecular modification type.
6. a kind of different efficient cross-film polymer micelle carrier of molecular modification type described in claim 5, react molten in preparation process Agent is water, methanol, n,N-Dimethylformamide, tetrahydrofuran, toluene, tetrahydrofuran, dichloromethane, dimethyl sulfoxide (DMSO), formyl Amine, the mixed solvent of water and methanol, the mixed solvent of water and n,N-Dimethylformamide, the mixed solvent of water and formamide, N, The mixed solvent of dinethylformamide and formamide.
7. the application of the efficient cross-film polymer micelle of different molecular modification type described in claim 1, it is characterised in that can be used for The injection of intravascular or intramuscular or oral, external application the carrier with pharmaceutical active or pharmacological activity molecule, it is such to carry medicine The molecule for learning activity or pharmacological activity is selected from：Non-steroid anti-inflammatory drug, steroidal anti-inflammatory drugs, suppress uric acid generation class gout suppressant Thing, promote uric acid discharge class anti-gout drugs and taxanes, flavonoids, camptothecin, vinca, Anthraquinones, ring spore Plain class, antifol, purine antagonist, Pyrimidine antagonists, any one in dihydropyridine series antineoplastic medicament or two Composition.
8. the application described in claim 7, is realized in the following ways：
A. the efficient cross-film polymer micelle carrier of different molecular modification type and water are dissolved by weight for 1~50: 1000 ratio, are obtained To nano micellar solution；
B. after therapeutically effective amount is dissolved with pharmaceutical active or pharmacological activity molecule with pharmaceutically acceptable organic solvent, Preformed solution is mixed to get with the nano micellar solution；By preformed solution after ultrasonic or high-pressure homogeneous processing, using dialysis, Ultrafiltration or evaporating organic solvent, obtain the polypeptide drug-loaded micelle solution that particle diameter is 10~1000nm；Or by preformed solution through rotation Turn evaporation after bottle wall forms film, redissolved with water or scattered, through ultrasonic or high-pressure homogeneous processing, be prepared particle diameter for 10~ The polypeptide drug-loaded micelle solution of 1000nm；
C. the polypeptide drug-loaded micelle solution being prepared, which is characterized in that, can also add pharmaceutically acceptable freeze drying protectant, and use is cold Freeze dry method and freeze-dried powder is made；The freeze drying protectant is lactose, glucose, mannitol, sodium chloride, amino acid, citric acid One or more in salt, acetate, phosphate.
9. a kind of different efficient cross-film polymer micelle of molecular modification type according to claim 1, it is characterised in that described Pharmaceutically acceptable organic solvent is：Methanol, ethanol, acetone, tetrahydrofuran, n,N-Dimethylformamide, dimethyl sulfoxide (DMSO) Deng organic solvent or their mixed solvent.
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CN108815552A (en) * 2018-07-05 2018-11-16 四川大学 A kind of drug controllably loads and the bio-medical coating material of long-acting slow-release and preparation method thereof
CN109528648A (en) * 2018-12-11 2019-03-29 中国药科大学 The amphipathy macromolecule prodrug micelle and its preparation method and application of the slightly sour environmental response of tumor stroma
CN109700779A (en) * 2018-09-14 2019-05-03 苏州科技大学 The degradable polymer microspheres of compound sulfhydryl modified chitosan
CN109925515A (en) * 2019-03-05 2019-06-25 上海珑欣生物医学科技有限公司 For treating microRNA nano-complex and its preparation and the application of colon cancer
CN112107542A (en) * 2020-09-03 2020-12-22 西北师范大学 Has tumor pH and H2O2Multifunctional polymer micelle with specific activated antitumor activity and preparation method thereof
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Cited By (7)

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Publication number Priority date Publication date Assignee Title
CN108815552A (en) * 2018-07-05 2018-11-16 四川大学 A kind of drug controllably loads and the bio-medical coating material of long-acting slow-release and preparation method thereof
CN109700779A (en) * 2018-09-14 2019-05-03 苏州科技大学 The degradable polymer microspheres of compound sulfhydryl modified chitosan
CN109528648A (en) * 2018-12-11 2019-03-29 中国药科大学 The amphipathy macromolecule prodrug micelle and its preparation method and application of the slightly sour environmental response of tumor stroma
CN109925515A (en) * 2019-03-05 2019-06-25 上海珑欣生物医学科技有限公司 For treating microRNA nano-complex and its preparation and the application of colon cancer
CN112107542A (en) * 2020-09-03 2020-12-22 西北师范大学 Has tumor pH and H2O2Multifunctional polymer micelle with specific activated antitumor activity and preparation method thereof
CN113563493A (en) * 2021-07-01 2021-10-29 蚌埠医学院 Hydrophobic polysaccharide and preparation method and application thereof
CN113563493B (en) * 2021-07-01 2022-06-24 蚌埠医学院 Hydrophobic polysaccharide and preparation method and application thereof

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