CN103705940A - Preparation and anti-tumor application of natural active drug-polysaccharide targeted compound - Google Patents
Preparation and anti-tumor application of natural active drug-polysaccharide targeted compound Download PDFInfo
- Publication number
- CN103705940A CN103705940A CN201310742883.9A CN201310742883A CN103705940A CN 103705940 A CN103705940 A CN 103705940A CN 201310742883 A CN201310742883 A CN 201310742883A CN 103705940 A CN103705940 A CN 103705940A
- Authority
- CN
- China
- Prior art keywords
- polysaccharide
- natural activity
- activity medicine
- reaction
- medicine
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Abstract
The invention relates to a natural active drug-polysaccharide targeted compound with antitumor activity and targeting property. A natural active drug is introduced onto a polysaccharide framework by chemical grafting, and physically mixed with a ligand distearoylphosphatidyl ethanolamine-polyethyleneglycol-anisoyl amine with targeted transmission capacity to form a natural active drug-polysaccharide targeted compound, and the natural active drug-polysaccharide targeted compound can be self-assembled in water to form a nano micelle. The method is characterized in that the natural active drug-polysaccharide targeted compound has targeted transmission capacity, enhances the tumor transfer efficiency of the preparation, reduces the untoward reaction and enhances the tolerance of the patient; the hydrophobic inner core formed by the hydrophobic group can physically wrap the antineoplastic drug, thereby obviously improving the water solubility of the antineoplastic drug; and the antineoplastic drug which is physically wrapped by the chemically coupled ursolic acid can implement combined treatment of tumors and lower the toxic or side effect. The preparation method is simple and easy to operate, has the advantage of higher yield, and can easily implement industrialization.
Description
Technical field
The invention belongs to field of pharmaceutical preparations, relate to a kind of preparation method of natural activity medicine-polysaccharide targeting complex and the application in antitumor thereof.
Background technology
In current global range, cancer has become frequently-occurring disease and the commonly encountered diseases of serious threat human health.World Health Organization (WHO) points out, cancer is captured more than 700 ten thousand people's life every year in the whole world at present, and this numeral also can continue to rise.Therefore for the treatment of cancer with study very urgently, be, new century mankind urgent need to solve the problems.
The features such as natural activity medicine is owing to having wide material sources, and biological activity is high, and toxic and side effects is few, have become the focus that people study in recent years.Its biological activity mainly comprises: antiinflammatory, antioxidation, blood fat reducing, blood pressure lowering, antitumor, antiviral etc.Wherein, its antineoplastic action is the focus of expert and scholar's research always.For example: gamlogic acid is one of effective ingredient of the secreted dry resin of Garcinia maingayii, for spectrum antineoplastic agent, kinds of tumors is shown to stronger anti-tumor activity, and the toxicity in effective dosage ranges is less, can optionally kill cancerous cell, and normal hemopoietic system and immunologic function are had no significant effect.There are some researches show that gamlogic acid has obvious dose-dependent inhibition effect to the propagation of human hepatoma cell strain SMMC-7721 and QGY-7701, and to a little less than normal person's hepatoma cell strain L-02 effect relatively.Curcumin is the active component extracting from Rhizoma Curcumae Longae, has pharmacological action widely, utilizes phenolic hydroxyl group Mulberry Extract, and radiation medicine liver damage, oxidative damage are shielded; By regulating cell cycle, cell death inducing, regulate gene expression to play antitumor action.Ursolic acid is pentacyclic triterpenoid, is distributed widely in occurring in nature, has antioxidation, antiinflammatory, protects the liver, the effect such as blood fat reducing.Discovered in recent years, ursolic acid can suppress the growth of Several Kinds of Malignancy cell.Research shows, ursolic acid has good inhibitory action to the propagation of tumor cell.Ursolic acid can inducing tumor cell generation apoptosis simultaneously.But most of natural activity medicines are insoluble in water, bioavailability is low, has greatly limited its formulation application.
Polysaccharide is the important component part of all Living organisms, is controlling cell division, and Cell differentiation inducing activity and the body homergy aspect that sustains life have great significance.In addition, because polysaccharide has good biocompatibility and biodegradability, backbone structure improves the advantages such as its physicochemical property owing to existing a large amount of active group (as carboxyl and amino etc.) to be easy to chemical modification, is more and more modified into amphiphilic polysaccharide conjugate.For example, hyaluronic acid is a kind of linear macromolecule mucopolysaccharide repeatedly alternately being connected and composed by the disaccharidase unit of glucuronic acid and 2-Acetamido-2-deoxy-D-glucose, there is good biocompatibility, biodegradability, high viscoelasticity, and can be combined with some specific receptor specificity of cell surface, therefore, the pharmaceutical carrier through hyaluronic acid transformation has the advantages such as good biocompatibility, targeting, slow-releasing and high drug load.Heparin and derivant thereof are by glucamine, L-idose aldehyde glycosides, N-Acetyl-D-glucosamine and D-Glucose aldehydic acid alternately form acid mucopolysaccharide sulfuric ester, there is good water solublity, because containing a large amount of hydroxyls and carboxyl in its structure, be easy to carry out structural modification.
Natural activity medicine-polysaccharide conjugate refers to small-molecule drug is connected on polysaccharide by covalent bond, as a part for side group.Its main advantage is: solid tumor is had to tendentiousness deposition, i.e. EPR effect; Prolong drug, at the circulation time of blood, is avoided engulfing of reticuloendothelial system; Improve hydrophobic drug solubility etc.
Along with the raising of Protocols in Molecular Biology, and from the molecular level of cell receptor and multiplication regulatory the further understanding to Tumorigenesis, people start to carry out to take the targeted therapy research that cell receptor etc. is target spot.Sigma-receptor is the embrane-associated protein of knowing, and psychosis is had to high-affinity.These expression of receptor are at normal tissue, liver for example, endocrine gland, kidney, lung, gonad, the central nervous system ovary of unifying.At a series of body tumor tissues overexpression sigma-receptor, melanoma for example, non-minicell pulmonary carcinoma, neurogenic breast tumor, and tumor of prostate.Benzoylamide shows the good affinity to sigma-receptor.Natural activity medicine-polysaccharide targeting complex is to utilize to have the ligand modified of targeting and form, and making to be modified the natural activity medicine-polysaccharide targeting complex obtaining can produce specifically and be combined with cell, increases its transport efficacy, reduces toxic and side effects.
Natural activity medicine-polysaccharide targeting complex can self assembly in aqueous solution, forms the outside hydrophilic micellar structure of inner hydrophobic, and its hydrophobic kernel can load hydrophobic medicine, increases thus the dissolubility of the hydrophobic drug being carried by bag.By chemical coupling and physically encapsulation antitumor drug, can reach the effect of co-therapy, improve curative effect.Therefore, natural activity medicine-polysaccharide targeting complex not only can be used alone as and prepare polymer drug, can also be as the carrier of pharmaceutical active or pharmacologically active molecule.
Summary of the invention
The object of the present invention is to provide a kind of natural activity medicine-polysaccharide targeting complex, by the method for chemical coupling, the natural activity medicine with anti-tumor activity is grafted on the skeleton of polysaccharide, form the amphipathic conjugate of natural activity medicine-polysaccharide, in water, be self-assembled into polymer micelle, again by the method for physical mixed, anisamide derivant-DSPE-PEG-anisamide (DSPE-PEG-AA) anchor of formed polymer micelle and sigma-receptor targeting is closed, form natural activity medicine-polysaccharide targeting complex polymer micelle.Because DSPE-PEG-AA has the effect of targeting sigma-receptor, so can make conjugate occur to be combined specifically with cell, increase the tumor transport efficacy of medicine, reduce the toxic and side effects of medicine, increase patient's toleration.
Another object of the present invention is to provide the preparation method of above-mentioned natural activity medicine-polysaccharide targeting complex.
Of the present invention also have an object to be to provide the application of above-mentioned natural activity medicine-polysaccharide targeting complex in antitumor.
In order to achieve the above object, the invention provides a kind of natural activity medicine-polysaccharide targeting complex, this complex is at the carboxyl of polysaccharide or through on the carboxyl of derivatization, by the method for chemical coupling, grafting natural activity medicine, form amphipathic natural activity medicine-polysaccharide conjugate, in water, self assembly forms polymer micelle.By the method for physical mixed, in conjunction with DSPE-PEG-AA, form natural activity medicine-polysaccharide target polymer micelle.This polymer micelle has improved stability and the dissolubility of natural activity medicine greatly, increase its initiatively targeted delivery characteristic, increased the bioavailability of active medicine, by chemical coupling and physically encapsulation antitumor drug, greatly reduce the toxic and side effects of medicine, increased patient's toleration.
Described natural activity medicine-polysaccharide targeting complex, wherein polysaccharide is selected from low molecular weight heparin, desulfated heparin, unfraction heparin, hyaluronic acid, alginic acid, chrondroitin, poly-sulfated chrondroitin, carboxymethyl chitosan, carboxyetbyl chitosan, quaternized carboxymethyl chitosan, N-octyl group-O, N-carboxymethyl chitosan, Sensor Chip CM 5 or carboxymethyl lentinan.
Described natural activity medicine-polysaccharide targeting complex, natural activity medicine wherein, be selected from terpene substances ursolic acid, enoxolone, oleanolic acid and triptolide, aldehydes matter honokiol and curcumin, Flavonoid substances gamlogic acid and baicalin and anthraquinones chrysophanic acid and resveratrol.
The preparation method of described natural activity medicine-polysaccharide targeting complex, different according to the structure of natural activity medicine, comprise following two methods:
1. in the structure of natural activity medicine, contain carboxyl
1. technique I: natural activity medicine is dissolved in appropriate solvent, take dicyclohexylcarbodiimide (DDC) and N-Hydroxysuccinimide (NHS) is activator, employing Diamines is linking arm, by amide, reacts, and obtains amidized natural activity medicine.
Technique II: part terpene substances, by oxidation reaction, changes into carbonyl by hydroxyl oxygen, and the ursolic acid of formed 3-carbonyl and oxammonium hydrochloride. are dissolved in pyridine, reaction generates white solid, this white solid and sodium cyanoborohydride (NaBH
3cN), sodium acetate (CH
3cOONa) and titanous chloride. (TiCl
3) be dissolved in methanol, reaction generates amidized natural activity medicine.
2. polysaccharide is dissolved in reaction dissolvent, 1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDC) of take is activator, by amide, is reacted natural activity medicine is grafted on polysaccharide skeleton.
3. technique I: the ratio that is 1～50: 1000 by weight by natural activity medicine-polysaccharide conjugate and water is dissolved, DSPE-PEG-AA and water are 1～50: 1000 ratio dissolving by weight, DSPE-PEG-AA solution is added dropwise in the amphipathic conjugate solution of natural activity medicine-polysaccharide, stirring at room, obtains natural activity medicine-polysaccharide targeting complex.
Technique II: natural activity medicine-polysaccharide conjugate is mixed in 1: 50～50: 1 ratios with DSPE-PEG-AA, dissolve with distilled water, stirring at room, obtains natural activity medicine-polysaccharide targeting complex.
2. in the structure of natural activity medicine, do not contain carboxyl
1. polysaccharide is dissolved in reaction dissolvent, the DMAP (DMAP) of take is catalyst, 1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDC) is dehydrant, first by after the activated carboxylic on polysaccharide, natural activity medicine is dissolved in to appropriate solvent, by esterification, natural activity medicine is grafted on polysaccharide.
2. technique I: the ratio that is 1～50: 1000 by weight by natural activity medicine-polysaccharide conjugate and water is dissolved, DSPE-PEG-AA and water are 1～50: 1000 ratio dissolving by weight, DSPE-PEG-AA solution is added dropwise in natural activity medicine-polysaccharide conjugate solution, stirring at room, obtains natural activity medicine-polysaccharide targeting complex.
Technique II: natural activity medicine-polysaccharide conjugate is mixed in 1: 50～50: 1 ratios with DSPE-PEG-AA, dissolve with distilled water, stirring at room, obtains natural activity medicine-polysaccharide targeting complex.
Described preparation method 1 and 2, wherein appropriate solvent is selected from the mixture of oxolane, DMF, dimethyl sulfoxide or oxolane and DMF or oxolane and dimethyl sulfoxide; Described reaction dissolvent is selected from the mixed solvent into water or Methanamide or DMF and water or Methanamide and water or Methanamide and oxolane or Methanamide and DMF.
Described preparation method 1, wherein linking arm is selected from the Alkylenediamine of p-phenylenediamine (PPD), cystamine or carbon number 2～12.
The technique II of the step of described preparation method 1 in 1., wherein part terpene substances comprises ursolic acid, oleanolic acid and enoxolone.
Described natural activity medicine-polysaccharide targeting complex, can be used alone as and prepare polymer drug, also can be used as the medicine that preparation bag carries hydrophobicity pharmaceutical active or pharmacologically active molecule; The medicine that prepared polymer drug or bag carry hydrophobicity pharmaceutical active or pharmacologically active molecule is injection, tablet, capsule, pill, granule, oral solution, patch, liniment, lotion, gel, varnish, ointment, spray, aerosol or nasal formulations.
Described pharmaceutical active or pharmacologically active molecule are selected from taxanes, camptothecin, vincristine class, fear quinones, podophillotoxines, amycin class or tretinoin antitumor drug.
Natural activity medicine-polysaccharide targeting complex of described carrying anti-tumor medicine, can be as injection, oral, external or mucosa delivery.Wherein drug administration by injection optimizing injection, freeze-dried powder, oral administration preferred tablet, capsule, pill, syrup, granule, oral solution, the preferred patch of topical administration, liniment, lotion, gel, varnish, ointment, the preferred spray of mucosa delivery, aerosol, nasal formulations.
The method operating procedure that this natural activity medicine-polysaccharide targeting complex is made medicament-carried nano micelle is as follows: natural activity medicine-polysaccharide targeting complex and water are 3～50: 1000 ratio dissolving by weight, obtain natural activity medicine-polysaccharide nano-micelle; By the indissoluble for the treatment of effective dose or be slightly soluble in the pharmaceutically acceptable dissolution with solvents of antitumor drug of water, after described natural activity medicine-polysaccharide nano-micelle mixes, through ultrasonic or high pressure homogenize, process, solution is removed organic solvent micromolecule by dialysis or ultrafiltration or post partition method and is obtained the nano-micelle that particle diameter is 10～1000nm, the nano-micelle obtaining can directly be applied, and also can recycle by lyophilization or dry the making after solid preparation redissolves of spraying.
Concrete scheme is as follows:
Containing carboxyl or through derivatization, forming on carboxylic polysaccharide molecule chain and to introduce the natural activity medicine with antitumor action, make it have amphipathic, in aqueous medium, can be self-assembled into nano-micelle, relatively hydrophobic natural activity medicine is agglomerated into kernel, polysaccharide molecule hydrophilic chain height of formation hydrophilic shell, has the anti-tumor activity of raising, stablizes micelle, effectively hides organism endothelium reticular system.By physical mixed DSPE-PEG-AA, form natural activity medicine-polysaccharide targeting complex, can be oriented to tumor cell, realize the targeted delivery of medicine.Therefore, this natural activity medicine-polysaccharide targeting complex is the pharmaceutical carrier that a class is good, especially for insoluble anti-tumor medicament.This natural activity medicine-polysaccharide targeting complex is as pharmaceutical carrier, and particle diameter is controlled at 10～1000nm, smooth surface, and good evenness, at good dispersion, drug loading is high.This natural activity medicine-polysaccharide targeting complex can be used for injection, oral, external or mucosa delivery.
The concrete preparation method synthetic and micelle of described natural activity medicine-polysaccharide targeting complex is as follows:
One, natural activity medicine-polysaccharide conjugate is synthetic
(1) in the structure of natural activity medicine, contain carboxyl
1. technique I: natural activity medicine is dissolved in suitable organic solvent, take dicyclohexylcarbodiimide and N-Hydroxysuccinimide as activator, control reaction 1 condition to reacting completely, sucking filtration, filtrate is precipitated with appropriate solvent 1, sucking filtration obtains precipitate, and vacuum drying obtains ester in the middle of the activity of natural activity medicine; The middle ester of natural activity medicine and Diamines linking arm are dissolved in to N by suitable molar ratio, in dinethylformamide, Alkylenediamine is slowly added drop-wise in the solution of ester in the middle of the activity of natural activity medicine, control reaction 2 to completely, with appropriate solvent 2, precipitate, precipitate carries out respectively pickling, washing, sucking filtration obtains precipitate, and vacuum drying obtains amidized natural activity medicine.
Technique II:A. part terpene substances is dissolved with dichloromethane, progressively splashes in the solution that is dissolved with oxidant stirring at room certain hour under uniform temperature.Reactant liquor after filtration, washing, washing, dry, filter, concentrated after, silica gel column chromatography eluting, concentrated, obtains solid.
B. the solid and the oxammonium hydrochloride. that reaction A are obtained are dissolved in pyridine, and certain hour refluxes under uniform temperature.Question response liquid is cooled to after room temperature, and distilled water wash, filtration, washing, dry, obtain solid.
C. the solid and sodium cyanoborohydride and the sodium acetate that reaction B are obtained, be dissolved in methanol, under nitrogen protection, reacts.Under condition of ice bath, slowly drip titanium trichloride solution, stirring at room certain hour.By the turbid solution of gained, add appropriate distilled water, revolve nor-alcohol, regulator solution pH is to neutral, and with dichloromethane extraction, organic layer washing, dry, filtration, filtrate are concentrated.Mixed solvent recrystallization with methanol, dichloromethane and petroleum ether, obtains solid, i.e. amidized natural activity medicine.
Synthetic route is as follows:
(R
1-COOH: ursolic acid, enoxolone, oleanolic acid, gamlogic acid or chrysophanic acid; H
2n-R
2-N
2h is the Alkylenediamine of p-phenylenediamine (PPD), cystamine or carbon number 2～12)
(R
1'-OH: ursolic acid, oleanolic acid and enoxolone)
2. polysaccharide molecule is dissolved in to reaction dissolvent, under nitrogen protection, 1-ethyl-(3-dimethylaminopropyl) carbodiimide of take is activator; add amidized natural activity medicine; control reaction to reacting completely, after reaction finishes, with appropriate solvent, precipitate; filtration is precipitated thing; redissolve, ultrasonic, dialysis; lyophilization, obtains the amphipathic conjugate of natural activity medicine-polysaccharide.
Synthetic route is as follows:
(R
3-COOH: low molecular weight heparin, desulfated heparin, unfraction heparin, hyaluronic acid, alginic acid, chrondroitin, poly-sulfated chrondroitin, carboxymethyl chitosan, carboxyetbyl chitosan, quaternized carboxymethyl chitosan, N-octyl group-O, N-carboxymethyl chitosan, Sensor Chip CM 5 or carboxymethyl lentinan)
3. technique I: the ratio that is 1～50: 1000 by weight by natural activity medicine-polysaccharide conjugate and water is dissolved, DSPE-PEG-AA and water are 1～50: 1000 ratio dissolving by weight, DSPE-PEG-AA solution is dripped and is slowly added dropwise in the amphipathic conjugate solution of natural activity medicine-polysaccharide, stirring at room reaction certain hour, obtains natural activity medicine-polysaccharide targeting complex.
Technique II: natural activity medicine-polysaccharide conjugate is mixed in 1: 50～50: 1 ratios with DSPE-PEG-AA, dissolve with distilled water, stirring at room certain hour, obtains natural activity medicine-polysaccharide targeting complex.
(2) in the structure of natural activity medicine, do not contain carboxyl
1. polysaccharide is dissolved in reaction dissolvent, take DMAP as catalyst, 1-ethyl-(3-dimethylaminopropyl) carbodiimide is dehydrant, under nitrogen protection, by the activated carboxylic on polysaccharide; Natural activity medicine is dissolved in to suitable solvent, is slowly added drop-wise in polysaccharide solution, control reaction to complete, question response finishes, and by appropriate solvent precipitation, filters to obtain precipitate, redissolves, ultrasonic, dialysis, and lyophilization, obtains natural activity medicine-polysaccharide conjugate.
Synthetic route is as follows:
(R
3-COOH: low molecular weight heparin, desulfated heparin, unfraction heparin, hyaluronic acid, alginic acid, chrondroitin, poly-sulfated chrondroitin, carboxymethyl chitosan, carboxyetbyl chitosan, quaternized carboxymethyl chitosan, N-octyl group-O, N-carboxymethyl chitosan, Sensor Chip CM 5 or carboxymethyl lentinan; R
4-OH is triptolide, baicalin, honokiol, curcumin or white reed Herba chenopodii alcohol.)
2. technique I: the ratio that is 3～50: 1000 by weight by the amphipathic conjugate of natural activity medicine-polysaccharide and water is dissolved, DSPE-PEG-AA and water are 1～50: 1000 ratio dissolving by weight, DSPE-PEG-AA solution is dripped and is slowly added dropwise in the amphipathic conjugate solution of natural activity medicine-polysaccharide, stirring at room reaction certain hour, obtains natural activity medicine-polysaccharide targeting complex.
Technique II: natural activity medicine-polysaccharide conjugate is mixed in 1: 50～50: 1 ratios with DSPE-PEG-AA, dissolve with distilled water, stirring at room certain hour, obtains natural activity medicine-polysaccharide targeting complex.
The concrete steps of step in described reaction (1) technique I are 1. as follows:
Described suitable organic solvent is selected from the mixture of oxolane, DMF, dimethyl sulfoxide or oxolane and DMF or oxolane and dimethyl sulfoxide.
Described control 1 condition of reacting is under nitrogen protection, and ice bath reaction 0.2～2h, rises to room temperature, reaction 6～72h.
Described precipitation solvent 1 is normal hexane.
The middle ester of described natural activity medicine and Alkylenediamine are 5: 1～80: 1 by suitable molar ratio.
Described control is reacted 2 conditions and is dripped speed at every interval 2～30 seconds for reaction; Reaction temperature is 0～40 ℃; After Diamines drips, then react 3～12h.
Described precipitation solvent 2 is saturated sodium-chloride.
Described pickling solvent is the rare HCl of 1mol/L, and washing solvent is ultra-pure water or distilled water.
The concrete steps of step in described reaction (1) technique II are 1. as follows:
Described part terpene substances comprises ursolic acid, oleanolic acid and enoxolone.
In described reaction A, the oxidant of oxidation reaction is pyridinium chlorochromate, and solvent is dichloromethane, or the oxidant of oxidation reaction is Jones reagent, and solvent is dichloromethane-acetone; Described temperature is controlled as-10～30 ℃, and the described stirring at room time is 0.5～10h.
In described reaction B, it is 80～150 ℃ that temperature is controlled; Described return time is 2～24h.
In described reaction C, the stirring at room time is 2～36h.
Step in described reaction (1) is 2. as follows with step concrete steps 3.:
Described step 2. in, reaction dissolvent is selected from the mixed solvent of water or Methanamide or DMF and water or Methanamide and water or Methanamide and oxolane or Methanamide and DMF; Described control is reacted for first adding after 1-ethyl-(3-dimethylaminopropyl) carbodiimide activator reaction 0.2～2h, then adds amidized natural activity medicine, room temperature reaction 6～72h; Described precipitation solvent is ice acetone, ice ether, ice ethanol or glacial acetic acid ethyl ester; Described dialysis time is 0.5～3d.
In described step technique I and technique II 3., the stirring at room response time is 10～60min.
In described reaction (2), concrete steps are as follows:
Described step 1. in, reaction dissolvent is the mixed solvent of water or Methanamide or DMF and water or Methanamide and water or Methanamide and oxolane or Methanamide and DMF; The mol ratio of described polysaccharide, catalyst, dehydrant is 1: 0.2～0.5: 3～10; The appropriate solvent of described dissolving natural activity medicine is the mixture of oxolane, DMF, dimethyl sulfoxide or oxolane and DMF or oxolane and dimethyl sulfoxide; Described control reaction, for to add catalyst and activator by above-mentioned mol ratio, after reaction 0.2～2h, adds natural activity medicine, and controlling a reaction speed is 2～30 seconds, every interval, room temperature reaction 6～72h again after dripping; Described suitable precipitation solvent is ice acetone, ice ether, ice ethanol or glacial acetic acid ethyl ester; Described dialysis time is 0.5～3d.
In described step technique I and technique II 2., stirring at room certain hour is 10～60min, is preferably 45min.
Two, the preparation method of natural activity medicine-polysaccharide targeting complexes micelle
In the ratio of dissolving natural activity medicine-polysaccharide targeting complex of 3～30mg in every 1mL water, natural activity medicine-polysaccharide targeting the complex making is soluble in water, through ultrasonic or high pressure homogenize, process, being prepared into particle diameter is natural activity medicine-polysaccharide targeting complexes micelle of 10～1000nm.
Three, using natural activity medicine-polysaccharide targeting complex as pharmaceutical carrier, preparation is dissolved with appropriate solvent as paclitaxel containing the medicine of insoluble anti-tumor medicament, mix with natural activity medicine-polysaccharide targeting compound water solution, through ultrasonic or high pressure homogenize, process, by dialysis or ultrafiltration or post partition method, remove organic solvent and micromolecule, make the nano-micelle that particle diameter is 10～1000nm.Described appropriate solvent, refers to the solvent that can dissolve this medicine pharmaceutically using.
The present invention has following advantage and effect with respect to prior art:
(1) the present invention forms amphipathic conjugate by natural activity medicine branch hydrophobic and that have an antitumor action to polysaccharide skeleton, water solublity and the stability of natural activity medicine have obviously been improved, extend its time in blood circulation, increased its bioavailability.Preparation process is simple, and processing ease reduces cost, raises the efficiency, and easily realizes industrialization.
(2) utilization of the present invention has the DSPE-PEG-AA of targeting, has realized the targeting of this system, has improved tumor transport efficacy, has reduced the toxic and side effects of medicine, has increased patient's toleration.Natural activity medicine-polysaccharide targeting the complex generating, is a kind of high molecular nanometer combination drug, is expected to become the superior nano combined medicine of a kind of antineoplastic.
(3) natural activity medicine-polysaccharide targeting complex provided by the invention, both can be used alone as polymer drug, also can be used as the carrier of pharmaceutical active or pharmacologically active molecule, had application space widely; In addition, the nano-micelle carrier forming by the self assembly of natural activity medicine-polysaccharide targeting complex, can realize the number of ways medicine carryings such as chemistry and physics, is a kind of pattern of very promising targeting combined chemotherapy.By chemical coupling natural activity medicine and other antitumor drug of physically encapsulation, can realize the complementation of combined chemotherapy mechanism of drug action, strengthen anti-tumor activity, reach the effect of synergy.
(4) natural activity medicine-polysaccharide targeting complex provided by the invention, not only has the good biological characteristics of polysaccharide molecule, can also bring into play better natural activity medicine antitumor, antioxidation, antiinflammatory, protect the liver, the effect such as blood fat reducing.
(5) the invention provides natural activity medicine-polysaccharide targeting complex and can be used for oral, injection, external or mucosa delivery, have tight security, particle diameter can be controlled in 10～1000nm.
Accompanying drawing explanation
Fig. 1: flow cytometer quantitative expedition natural activity medicine-polysaccharide targeting complex cellular uptake.
Fig. 2-1,2-2,2-3:MTT method are measured the anti-tumor activity of natural activity medicine-polysaccharide targeting complex.
Fig. 3-1,3-2,3-3,3-4: natural activity medicine-polysaccharide targeting complex is to different growth of tumour cell and antitumor action.
Fig. 4-1,4-2,4-3,4-4: the impact that natural activity medicine-polysaccharide targeting complex forms HUVEC tubule.
Fig. 5-1,5-2:MTT method are measured the anti-tumor activity of natural activity medicine-polysaccharide targeting complex of carrying anti-tumor medicine.
The specific embodiment
Below by embodiment, to the present invention's further instruction in addition, but following embodiment does not limit the interest field of this patent.
Embodiment 1: the preparation of curcumin-Sensor Chip CM 5 targeting complex
Getting 0.2mol Sensor Chip CM 5 adds in 15ml water, magnetic agitation is to dissolving completely, with acetic acid, regulate left and right, PH to 6～7, add again 0.6mol propane diamine, after mix homogeneously, add again 0.2mol1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDC) and 0.3mol N-Hydroxysuccinimide (NHS), room temperature reaction 12h, dialysis 2d, must the dissociate Sensor Chip CM 5 intermediate of one end amino of lyophilization.Getting above-mentioned intermediate is scattered in appropriate methanol, the methanol solution that adds the curcumin of 0.9mol, ultrasonic back flow reaction 2.5h, absolute methanol for crude product, absolute ether, dichloromethane cyclic washing are colourless to filtrate, and vacuum drying obtains the amphipathic conjugate of curcumin-Sensor Chip CM 5 after purification.4mg curcumin-Sensor Chip CM 5 conjugate is dissolved in 4mL distilled water, and 2mg DSPE-PEG-AA is dissolved in 2mL distilled water.The DSPE-PEG-AA solution of 600 μ L is slowly added dropwise to curcumin-Sensor Chip CM 5 conjugate solution, stirs 45min, obtain curcumin-Sensor Chip CM 5 targeting complex.
Embodiment 2: the preparation of ursolic acid-low molecular weight heparin targeting complex
300mg ursolic acid dissolves with dichloromethane, and 10 ℃ progressively splash in the dichloromethane solution that is dissolved with 280mg pyridinium chlorochromate.After dropwising, stirring at room 1.5h.Reactant liquor after filtration, washing, washing, dry, filter, concentrated after, silica gel column chromatography, eluting, concentrated, obtains white solid.Get this white solid 59mg and 80mg oxammonium hydrochloride. and be dissolved in 7ml pyridine, above connect drying tube, 115 ℃ of about 6h that reflux.Reactant liquor is cooled to after room temperature, and distilled water wash filters, and washing is dry, obtains white solid.Get this white solid 120mg, 1.05g sodium cyanoborohydride, 1.05g sodium acetate, is dissolved in methanol, and reaction system is in N
2under protection, air in eliminating system.System is placed in ice bath, 0.8ml TiCl
3solution is slowly added drop-wise in reactant liquor.After dropwising, stirring at room 14h.The white opacity liquid of gained is added to appropriate distilled water, revolve nor-alcohol, with the NaOH solution of 2M, regulate pH to neutral, with dichloromethane extraction, organic layer washing, dry, to filter, filtrate is concentrated.With methanol, dichloromethane and petroleum ether mixed solvent recrystallization, obtain white solid, be amidized ursolic acid.Take 0.3mol low molecular weight heparin and be dissolved in formamide solution, 1.2mol1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDC) is dissolved in Methanamide.At N
2under protection, EDC is slowly added in low molecular weight heparin solution, after activation 15min, the amidized ursolic acid of 0.9mol is joined in the solution of low molecular weight heparin.Room temperature reaction 24h.After stopped reaction, add excessive cold acetone precipitated product, sucking filtration, obtains.Product redissolves with ultra-pure water, ice bath, and Probe Ultrasonic Searching 30min, the 2d (MWCO=3500) that dialyses in distilled water, crosses 0.8 μ m microporous filter membrane, and lyophilization, obtains product, the amphipathic conjugate of ursolic acid-low molecular weight heparin.The amphipathic conjugate of 3mg ursolic acid-low molecular weight heparin is dissolved in 3mL distilled water, and 2mg DSPE-PEG-AA is dissolved in 4mL distilled water.The DSPE-PEG-AA solution of 888 μ L is slowly added dropwise in the amphipathic conjugate solution of ursolic acid-low molecular weight heparin, stirs 30min, obtain ursolic acid-low molecular weight heparin targeting complex.
Embodiment 3: the preparation of gamlogic acid-hyaluronic acid targeting complex
Take gamlogic acid (GA) 0.6mol and be placed in eggplant-shape bottle, add the oxolane of 30mL, ice bath, at logical N
2condition under, add successively dicyclohexylcarbodiimide (DCC) and N-Hydroxysuccinimide (NHS), the mol ratio of GA, DCC, NHS is followed successively by 1: 1.5: 1.2.After ice bath reaction 30min, move to room temperature reaction 24h.Vacuum filtration disgorging DCU, obtains filtrate.Filtrate, with the normal hexane precipitation 12h of 3 times of amounts, is filtered, and vacuum drying, obtains succinimido GA.Take succinimido GA and dissolve with DMF, be slowly added drop-wise in 2mL cystamine, 55min drips off.React 8h, saturated common salt water precipitation, filters, and obtains the amidized GA of yellow mercury oxide, carries out successively pickling, washing, vacuum drying, 4 ℃ of preservations again.Take 0.2mol hyaluronic acid and be dissolved in DMF, add 0.3mol1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDC).At N
2under protection, EDC is slowly added in hyaluronic acid solution, after activation 15min, amidized GA is joined in hyaluronic solution.Room temperature reaction 24h.After stopped reaction, add excessive cold acetone precipitated product, sucking filtration, obtains.Product redissolves with ultra-pure water, ice bath, and Probe Ultrasonic Searching 30min, the 2d (MWCO=3500) that dialyses in distilled water, crosses 0.8 μ m microporous filter membrane, and lyophilization, obtains product gamlogic acid-hyaluronic acid amphipathic conjugate.4mg gamlogic acid-hyaluronic acid amphipathic conjugate is dissolved in 4mL distilled water, and 2mg DSPE-PEG-AA is dissolved in 2mL distilled water.The DSPE-PEG-AA solution of 500 μ L is slowly added dropwise in gamlogic acid-hyaluronic acid amphipathic conjugate solution, stirs 30min, obtain gamlogic acid-hyaluronic acid targeting complex.
Embodiment 4: the preparation of baicalin-carboxymethyl chitosan targeting complex
Take carboxymethyl chitosan 0.05mmol and be placed in eggplant-shape bottle, add 20mL Methanamide, heating, magnetic agitation makes its dissolving, be cooled to room temperature, add respectively catalyst DMAP (DMAP) 0.01mmol, dehydrant 1-ethyl-(3-dimethylaminopropyl) carbodiimide (EDC) 0.6mmol, activates after 30min under room temperature condition, adds baicalin 0.3mmol, N with 20mL, dinethylformamide dissolves, and is placed in constant pressure funnel, controls reaction and drips speed, 1～3h dropwises, room temperature reaction 24h.After TLC monitoring reaction extremely completely, after stopped reaction, add the ice acetone precipitation product of excess volume, sucking filtration, obtains product.Product redissolves with ultra-pure water, ice bath, and Probe Ultrasonic Searching 30min, the 2d (MWCO=3500) that dialyses in distilled water, crosses 0.8 μ m microporous filter membrane, and lyophilization, obtains the amphipathic conjugate of product baicalin-carboxymethyl chitosan.The amphipathic conjugate of 4mg baicalin-carboxymethyl chitosan is dissolved in 4mL distilled water, and 2mg DSPE-PEG-AA is dissolved in 2mL distilled water.The DSPE-PEG-AA solution of 700 μ L is slowly added dropwise to the amphipathic conjugate solution of baicalin-carboxymethyl chitosan, stirs 30min, obtain baicalin-carboxymethyl chitosan targeting complex.
Embodiment 5: preparation and the sign of natural activity medicine-polysaccharide targeting complex nano-micelle
1, the preparation of natural activity medicine-polysaccharide targeting complex nano-micelle: according to the character of natural activity medicine, whether consider needs under the condition of lucifuge, natural activity medicine-polysaccharide targeting complex 18mg is dissolved in 3m1 water, in stirring at room 30min, then after ultrasonic under ice bath or high pressure homogenize, 0.8 μ m membrane filtration, obtains.
2, by the 1 natural activity medicine-polysaccharide targeting complex nano-micelle preparing, get 1mL and be diluted with water to 3mL, with particle instrument (Malvern Instruments, Malvern, UK), measure, the results are shown in Table 1.
The particle diameter (nm) of natural activity medicine-polysaccharide targeting complex of table 1 different mixing proportion
A:DSPE-PEG-AA B: natural activity medicine-polysaccharide conjugate
Embodiment 6:: the Preparation and characterization that comprises paclitaxel natural activity medicine-polysaccharide targeting complex self-assembled nano micelle compositions
1, preparation technology
(1) dialysis
By natural activity medicine-polysaccharide targeting complex 18mg, be dissolved in 3m1 distilled water and stir 1h.Paclitaxel 10mg is dissolved in ethanol (methanol, acetonitrile).Then both mix, after Probe Ultrasonic Searching 30min, and distilled water dialysed overnight, centrifugal (3000rpm) 15min, with 0.8 μ m membrane filtration, lyophilization.
(2) emulsion-solvent evaporation method
By natural activity medicine-polysaccharide targeting complex 18mg, be dissolved in 3ml distilled water and stir 1h.Paclitaxel 10mg is dissolved in dichloromethane.Then the two mixes, Probe Ultrasonic Searching 30min, and the uncovered stirring of room temperature is spent the night, and makes dichloromethane volatilization, and centrifugal (3000rpm) 15min, with 0.8 μ m membrane filtration, lyophilization.
2, the mensuration of content of taxol in natural activity medicine-polysaccharide targeting complex self-assembled nano micelle
By HPLC (LC-2010C, Shimadzu, Japan) method, carry out assay.Mobile phase is methanol: water=75: 25 (v/v), and chromatographic column is Lichrospher C18 (150 * 4.6 μ m), pillar particle diameter is 5 μ m posts.Flow velocity is 1.0mL/min, and detection wavelength is 227nm (SPD-10A, UVdetector, Shimadzu, Japan), and column temperature is 30 ℃, and injected sample volume is 20 μ l.Drug loading with formula (1) calculation sample.The results are shown in Table 2.
Table 2 is loaded with natural activity medicine-polysaccharide targeting complex self-assembled nano micelle of paclitaxel
Embodiment 7: the Preparation and characterization that comprises amycin natural activity medicine-polysaccharide targeting complex self-assembled nano micelle compositions
1, preparation technology
Natural activity medicine-polysaccharide targeting complex 18mg, is dissolved in 3mL distilled water and stirs 1h.Amycin 9mg is dissolved in dimethyl sulfoxide (DMF).Then the two mixes, after Probe Ultrasonic Searching 30min, and distilled water dialysed overnight, centrifugal (3000rpm) 15min, with 0.8 μ m membrane filtration, lyophilization.
2, the mensuration of amycin content in natural activity medicine-polysaccharide targeting complex self-assembled nano micelle
With ultraviolet spectrophotometry, in 480nm wavelength place, measure the content of amycin.Drug loading with formula (1) calculation sample.The results are shown in Table 3.
Table 3 is loaded with natural activity medicine-polysaccharide targeting complex self-assembled nano micelle of amycin
Embodiment 8: the Preparation and characterization that comprises hydroxy camptothecin natural activity medicine-polysaccharide targeting complex self-assembled nano micelle
1, preparation technology
Natural activity medicine-polysaccharide targeting complex 20mg, is dissolved in 3mL distilled water and stirs 1h.Hydroxy camptothecin 10mg is dissolved in DMF (dimethyl sulfoxide).Then the two mixes, after Probe Ultrasonic Searching 30min, and distilled water dialysed overnight, centrifugal (3000rpm) 15min, with 0.8 μ m membrane filtration, lyophilization.
2, the mensuration of natural activity medicine-polysaccharide targeting complex self-assembled nano micelle hydroxy camptothecin content
With high performance liquid chromatogram-fluorescence detection method, carry out assay.Mobile phase is citrate buffer: acetonitrile: 75nmol/ml potassium dihydrogen phosphate=70: 23: 7 (v/v), chromatographic column is Lichrospher C18 (150 * 4.6 μ m), pillar particle diameter is 5 μ m.Flow velocity is 1.0ml/min, and fluoroscopic examination wavelength is λ
ex363nm and λ
ex530nm, column temperature is 50 ℃, injected sample volume is 20 μ l.Drug loading with formula (1) calculation sample.The results are shown in Table 4.
Table 4 is loaded with natural living-article medicine-polysaccharide targeting complex self-assembled nano micelle of hydroxy camptothecin
Embodiment 9: flow cytometer quantitative expedition natural activity medicine-polysaccharide targeting complex cellular uptake
B16 cell is inoculated in to 6 orifice plates, grows up to monolayer adherence cell after hatching 24h, suck culture fluid, use serum-free medium rinsing, add respectively (a) the coumarin 6 solution that dissociates; (b) carry ursolic acid-low molecular weight heparin targeting complex solution of coumarin 6; (c) carry ursolic acid-low molecular weight heparin conjugate solution of coumarin 6, hatch respectively 6h for 37 ℃.Afterwards, with the rapid peptic cell of 0.25% trypsin, with pre-cooling PBS, neutralize and blow and beat into cell suspension.Centrifugal 5min under 3000rpm, cell colony is with pre-cooling PBS rinsing and be made into individual cells suspension.Fluorescence intensity with the coumarin 6 of cells were tested by flow cytometry cellular uptake.The results are shown in Figure 1.Result shows, with the ursolic acid-low molecular weight heparin conjugate solution phase ratio that carries coumarin 6, the average fluorescent strength that carries ursolic acid-low molecular weight heparin targeting complex solution of coumarin 6 significantly strengthens, and shows that cell obviously increases carrying the picked-up of ursolic acid-low molecular weight heparin targeting complex of coumarin 6.
Embodiment 10:MTT method is measured the anti-tumor activity of natural activity medicine-polysaccharide targeting complex
By the B16F10 in exponential phase, MCF-7 and HepG2 cell digest with 0.02%EDTA, make cell suspension, respectively with 1 * 10
5/ ml cell concentration adds in 96 hole ELISA Plate, and every hole 100 μ l establish five multiple holes, put 37 ℃ of 5%CO
2in incubator, cultivate 24h left and right.The various samples with culture medium dilution that add respectively variable concentrations: (a1) ursolic acid; (b1) ursolic acid-unfraction heparin targeting complex; (a2) chrysophanic acid; (b2) chrysophanic acid-carboxyetbyl chitosan targeting complex; (a3) triptolide; (b3) triptolide-chrondroitin targeting complex.The concentration of the preparation after dilution is respectively 10 μ g/mL, hatch after 72h, discard whole supernatant, with PBS washing once, add blue (MTT, 5mg/mL) the 10 μ L of tetramethyl azo azoles, hatch 4h for 37 ℃, abandoning supernatant, each hole adds 150 μ LDMSO to dissolve the crystallization of first a ceremonial jade-ladle, used in libation in the mensuration absorbance (A of enzyme connection instrument 570nm wavelength place
test).And measure in the same way the trap of blank group (acellular strain) and matched group (n.s), be designated as respectively A
blankand A
control, by formula (2), calculate cell survival rate, the cytotoxicity of assess sample.The results are shown in Figure 2-1,2-2,2-3.
Result shows, compares natural drug, and the anti tumor activity in vitro to different tumor cells of natural drug-polysaccharide targeting complex obviously strengthens.
Embodiment 11: natural activity medicine-polysaccharide targeting complex is to different growth of tumour cell and antitumor action
By B16F10 tumor-bearing mice, OS-RC-2 tumor-bearing mice, MCF-7 tumor-bearing mice and HepG2 tumor-bearing mice are divided into respectively 5 groups at random, and 5/group, grouping and dosage are as follows: (a1) ursolic acid; (b1) ursolic acid-low molecular weight heparin conjugate; (c1) ursolic acid-low molecular weight heparin targeting complex; (d1) 5% glucose blank group; (a2) baicalin; (b2) baicalin-carboxymethyl chitosan carbohydrate conjugates; (c2) baicalin-carboxymethyl chitosan targeting complex; (d2) 5% glucose blank group; (a3) gamlogic acid; (b3) gamlogic acid-hyaluronic acid conjugate; (c3) gamlogic acid-hyaluronic acid targeting complex; (d3) 5% glucose blank group; (a4) curcumin; (b4) curcumin-Sensor Chip CM 5; (c4) curcumin-Sensor Chip CM 5 targeting complex; (d4) 5% glucose blank group.(a1) (b1), be (c1), 25mg/kg (in ursolic acid) with (d1) organizing dosage; (a2) (b2), be (c2), 10mg/kg (in baicalin) with (d2) organizing dosage; (a3) (b3), be (c3), 8mg/kg (in gamlogic acid) with (d3) organizing dosage; (a4) (b4), be (c4), 10mg/kg (in curcumin) with (d4) organizing dosage.According to the mode of administration every other day, with first day administration, count the 0th day, according to above-mentioned dosage, respectively at 0,2,4,6 days through above-mentioned group of preparation of tail vein injection.Within after drug withdrawal the 2nd day, put to death and dissect mice, peel off tumor, take tumor weight, according to formula (3), calculate tumor growth inhibition percentage.
The results are shown in Figure 3-1,3-2,3-3,3-4, result shows, compare matched group, natural activity medicine-polysaccharide conjugate has obvious enhancing from natural activity medicine-polysaccharide targeting complex to the anti-tumor activity of different tumor model mices, compare with natural activity medicine-polysaccharide conjugate, the anti-tumor activity of natural activity medicine-polysaccharide targeting complex is significantly increased.
The impact that embodiment 12-natural activity medicine-polysaccharide targeting complex forms HUVEC tubule
Get liquid Matrigel glue 50 μ L at 4 ℃ and add 96 orifice plates, tiling is placed on 1h in 37 ℃ of incubators and fixes, by Human umbilical vein endothelial cells (HUVEC) cell suspension 50 μ L (3 * 10
5individual/ml) join in Matrigel glue, with endotheliocyte serum-free medium, macromolecular drug complex is fitted on to desired concn, join in culture plate, every hole adds 50 μ L, after jiggling evenly, and at 37 ℃, 5%CO
2under condition, cultivate 8h, choose at random 3 visuals field, under 100 times of mirrors, repeated observation is taken pictures, and calculates tubule under each visual field and forms number.The concentration of laboratory sample group medicine is 100 μ g/mL.Negative control group is set simultaneously and only contains 5% hyclone.Application of sample group and matched group are all established 3 multiple holes.Laboratory sample group is (a1) low molecular weight heparin; (a2) N-desulfuration acidify heparin; (a3) heparin; (a4) N-O-desulfuration acidify heparin; (b1) ursolic acid; (b2) curcumin; (b3) triptolide; (b4) gamlogic acid; (c1) ursolic acid-low molecular weight heparin targeting complex; (c2) curcumin-N-desulfuration acidify heparin targeting complex; (c3) triptolide-heparin targeting complex; (c4) gamlogic acid-N-O-desulfuration acidify heparin targeting complex.The results are shown in Figure 4-1,4-2,4-3,4-4.As seen from the figure, natural activity medicine-polysaccharide targeting complex and matched group phase specific energy obviously suppress the formation of tubule, compare to suppress the ability that tubule forms and strengthen with free natural activity medicine or heparin and derivant thereof.Natural activity medicine-polysaccharide targeting the complex of proof in the present invention can suppress angiogenesis, and compares with free natural activity medicine or heparin and derivant thereof and suppress the ability that tubule forms and strengthen.
Embodiment 13:MTT method is measured the anti-tumor activity of natural activity medicine-polysaccharide targeting complex of carrying anti-tumor medicine
MCF-7 cell in exponential phase is digested with 0.02%EDTA, make cell suspension, respectively with 1 * 10
5/ ml cell concentration adds in 96 hole ELISA Plate, and every hole 100 μ l establish five multiple holes, put 37 ℃ of 5%CO
2in incubator, cultivate 24h left and right.The various samples with culture medium dilution that add respectively variable concentrations: (a1) paclitaxel; (a2) ursolic acid-low molecular weight heparin targeting complex that comprises paclitaxel; (b1) amycin; (b2) baicalin-carboxymethyl chitosan targeting complex that comprises amycin.The concentration of the preparation after dilution is respectively 0.01,0.1,1,10,100 μ g/mL, hatch after 72h, discard whole supernatant, with PBS washing once, add the blue (MTT of tetramethyl azo azoles, 5mg/mL) 10 μ L, hatch 4h for 37 ℃, abandoning supernatant, each hole adds 150 μ L DMSO to dissolve the crystallization of first a ceremonial jade-ladle, used in libation in the mensuration absorbance (A of enzyme connection instrument 570nm wavelength place
test).And measure in the same way the trap of blank group (acellular strain) and matched group (n.s), be designated as respectively A
blankand A
control, by formula (1), calculate cell survival rate, the cytotoxicity of assess sample.The results are shown in Figure 5-1,5-2.Result shows, compares antitumor drug, and the anti-tumor activity of natural drug-polysaccharide targeting complex of carrying anti-tumor medicine is stronger.
Claims (10)
1. natural activity medicine-polysaccharide targeting complex; it is characterized in that this natural activity medicine-polysaccharide targeting complex prepares by following method: the method by the natural activity medicine with anti-tumor activity by chemical coupling is grafted on the skeleton of polysaccharide; with part-DSPE-PEG-anisamide (DSPE-PEG-AA) physical mixed with targeted delivery ability, form natural activity medicine-polysaccharide targeting complex.
2. natural activity medicine-polysaccharide targeting complex according to claim 1, it is characterized in that: described natural activity medicine is selected from terpene substances ursolic acid, enoxolone, oleanolic acid and triptolide, aldehydes matter honokiol and curcumin, Flavonoid substances gamlogic acid and baicalin, and fear quinones substance chrysophanic acid and resveratrol; Described polysaccharide is selected from low molecular weight heparin, desulfated heparin, unfraction heparin, hyaluronic acid, alginic acid, chrondroitin, poly-sulfated chrondroitin, carboxymethyl chitosan, carboxyetbyl chitosan, quaternized carboxymethyl chitosan, N-octyl group-O, N-carboxymethyl chitosan, Sensor Chip CM 5 or carboxymethyl lentinan.
3. natural activity medicine-polysaccharide targeting complex according to claim 1, is characterized in that the preparation of this natural activity medicine-polysaccharide targeting complex comprises the steps:
(1) in the structure of natural activity medicine, contain carboxyl
1. technique I: natural activity medicine is dissolved in suitable organic solvent, take dicyclohexylcarbodiimide and N-Hydroxysuccinimide as activator, control reaction 1 condition to reacting completely, sucking filtration, filtrate is precipitated with appropriate solvent 1, sucking filtration obtains precipitate, and vacuum drying obtains ester in the middle of the activity of natural activity medicine; The middle ester of natural activity medicine and Diamines linking arm are dissolved in to N by suitable molar ratio, in dinethylformamide, Diamines linking arm is slowly added drop-wise in the solution of ester in the middle of the activity of natural activity medicine, control reaction 2 to completely, with appropriate solvent 2, precipitate, precipitate carries out respectively pickling, washing, sucking filtration obtains precipitate, and vacuum drying obtains amidized natural activity medicine;
Technique II:A. part terpene substances is dissolved with dichloromethane, progressively splashes in the solution that is dissolved with oxidant stirring at room certain hour under uniform temperature.After filtration, washing, washes reactant liquor, dry, filter, and after concentrating, silica gel column chromatography eluting, concentrated, obtain solid;
B. the white solid and the oxammonium hydrochloride. that reaction A are obtained are dissolved in pyridine, and certain hour refluxes under uniform temperature.Question response liquid is cooled to after room temperature, and distilled water wash filters, and washing is dry, obtains solid;
C. the solid and sodium cyanoborohydride and the sodium acetate that reaction B are obtained, be dissolved in methanol, under nitrogen protection, reacts.Under condition of ice bath, slowly drip titanium trichloride solution, stirring at room certain hour.By the turbid solution of gained, add appropriate distilled water, revolve nor-alcohol, regulator solution pH is to neutral, with dichloromethane extraction, organic layer washing, dry, filter, filtrate is concentrated, and the mixed solvent recrystallization with methanol, dichloromethane and petroleum ether, obtains solid, i.e. amidized natural activity medicine;
2. polysaccharide molecule is dissolved in to reaction dissolvent, under nitrogen protection, 1-ethyl-(3-dimethylaminopropyl) carbodiimide of take is activator, add amidized natural activity medicine, control reaction to reacting completely, after reaction finishes, with appropriate solvent, precipitate, filtration is precipitated thing, redissolve, ultrasonic, dialysis, lyophilization, obtains natural activity medicine-polysaccharide conjugate;
3. technique I: the ratio that is 1～50: 1000 by weight by natural activity medicine-polysaccharide conjugate and water is dissolved, DSPE-PEG-AA and water are 1～50: 1000 ratio dissolving by weight, DSPE-PEG-AA solution is slowly added dropwise in natural activity medicine-polysaccharide conjugate solution, stirring at room reaction certain hour, obtains natural activity medicine-polysaccharide targeting complex;
Technique II: natural activity medicine-polysaccharide conjugate is mixed in 1: 50～50: 1 ratios with DSPE-PEG-AA, dissolve with distilled water, stirring at room certain hour, obtains natural activity medicine-polysaccharide targeting complex;
(2) in the structure of natural activity medicine, do not contain carboxyl
1. polysaccharide is dissolved in reaction dissolvent, take DMAP as catalyst, 1-ethyl-(3-dimethylaminopropyl) carbodiimide is dehydrant, under nitrogen protection, by the activated carboxylic on natural polysaccharide; Natural activity medicine is dissolved in to suitable solvent, is slowly added drop-wise in polysaccharide solution, control reaction to complete, question response finishes, and by appropriate solvent precipitation, filters to obtain precipitate, redissolves, ultrasonic, dialysis, and lyophilization, obtains natural activity medicine-polysaccharide conjugate;
2. technique I: the ratio that is 3～50: 1000 by weight by the amphipathic conjugate of natural activity medicine-polysaccharide and water is dissolved, DSPE-PEG-AA and water are 1～50: 1000 ratio dissolving by weight, DSPE-PEG-AA solution is dripped and is slowly added dropwise in the amphipathic conjugate solution of natural activity medicine-natural polysaccharide, stirring at room reaction certain hour, obtains natural activity medicine-polysaccharide targeting complex;
Technique II: natural activity medicine-polysaccharide conjugate is mixed in 1: 50～50: 1 ratios with DSPE-PEG-AA, dissolve with distilled water, stirring at room certain hour, obtains natural activity medicine-polysaccharide targeting complex.
4. the preparation of natural activity medicine-polysaccharide targeting complex according to claim 3, wherein the technique I of the step of reaction (1) in is 1. characterised in that:
Described suitable organic solvent is selected from the mixture of oxolane, DMF, dimethyl sulfoxide or oxolane and DMF or oxolane and dimethyl sulfoxide;
Described control 1 condition of reacting is under nitrogen protection, and ice bath reaction 0.2～2h, rises to room temperature, reaction 6～72h;
Described precipitation solvent 1 is normal hexane;
It is 2～12 Alkylenediamine that described Diamines linking arm is selected from p-phenylenediamine (PPD), cystamine or carbon number;
The middle ester of described natural activity medicine and Alkylenediamine are 5: 1～80: 1 by suitable molar ratio;
Described control is reacted 2 conditions and is dripped speed at every interval 2～30 seconds for reaction; Reaction temperature is 0～40 ℃; After Diamines drips, then react 3～12h;
Described precipitation solvent 2 is saturated sodium-chloride;
Described pickling solvent is the rare HCl of 1mol/L, and washing solvent is ultra-pure water or distilled water.
5. the preparation of natural activity medicine-polysaccharide targeting complex according to claim 3, wherein the technique II of the step of reaction (1) in is 1. characterised in that:
Described part terpene substances comprises ursolic acid, oleanolic acid and enoxolone;
In described reaction A, the oxidant of oxidation reaction is pyridinium chlorochromate, and solvent is dichloromethane, or the oxidant of oxidation reaction is Jones reagent, and solvent is dichloromethane-acetone; Described temperature is controlled as-10～30 ℃; The described stirring at room time is 0.5～10h;
In described reaction B, it is 80～150 ℃ that temperature is controlled; Described return time is 2～24h;
In described reaction C, the stirring at room time is 2～36h.
6. the preparation of natural activity medicine-polysaccharide targeting complex according to claim 3, wherein 3. the step of reaction (1) is 2. characterised in that with step:
Described step 2. in, reaction dissolvent is selected from the mixed solvent into water or Methanamide or DMF and water or Methanamide and water or Methanamide and oxolane or Methanamide and DMF; Described control is reacted for first adding after 1-ethyl-(3-dimethylaminopropyl) carbodiimide activator reaction 0.2～2h, then adds amidized natural activity medicine, room temperature reaction 6～72h; Described precipitation solvent is ice acetone, ice ether, ice ethanol or glacial acetic acid ethyl ester; Described dialysis time is 0.5～3d;
In described step technique I and technique II 3., the stirring at room response time is 10～60min;
7. the preparation of natural activity medicine-polysaccharide targeting complex according to claim 3, is wherein characterised in that in reaction (2):
Described step 1. in, reaction dissolvent is the mixed solvent of Methanamide or Methanamide and oxolane or Methanamide and DMF; The mol ratio of described polysaccharide, catalyst, dehydrant is 1: 0.2～0.5: 3～20; The appropriate solvent of described dissolving natural activity medicine is the mixture of oxolane, DMF, dimethyl sulfoxide or oxolane and DMF or oxolane and dimethyl sulfoxide; Described control reaction, for to add catalyst and activator by above-mentioned mol ratio, after reaction 0.2～2h, adds natural activity medicine, and controlling a reaction speed is 2～30 seconds, every interval, room temperature reaction 6～72h again after dripping; Described suitable precipitation solvent is ice acetone, ice ether, ice ethanol or glacial acetic acid ethyl ester; Described dialysis time is 0.5～3d;
In described step technique I and technique II 2., stirring at room certain hour is 10～60min, is preferably 45min.
8. natural activity medicine-polysaccharide targeting complex claimed in claim 1, is characterized in that it can be used alone as and prepares polymer drug, or as preparation bag, carry the medicine of hydrophobicity pharmaceutical active or pharmacologically active molecule; The medicine that prepared polymer drug or bag carry hydrophobicity pharmaceutical active or pharmacologically active molecule is injection, tablet, capsule, pill, granule, oral solution, patch, liniment, lotion, gel, varnish, ointment, spray, aerosol or nasal formulations.
9. the application of natural activity medicine-polysaccharide targeting complex according to claim 8, is characterized in that described pharmaceutical active or pharmacologically active molecule are selected from: taxanes, camptothecin, vincristine class, fear quinones, podophillotoxines, amycin class or tretinoin antitumor drug.
10. according to the requirement of claim 9, it is characterized in that natural activity medicine-polysaccharide targeting complex prepares the method for medicament-carried nano micelle and comprise the steps: that natural activity medicine-polysaccharide and water are that 3～50: 1000 ratio is dissolved by weight, obtain natural activity medicine-polysaccharide nano-micelle; By the indissoluble for the treatment of effective dose or be slightly soluble in the pharmaceutically acceptable dissolution with solvents of antitumor drug of water, after described natural activity medicine-polysaccharide nano-micelle mixes, through ultrasonic or high pressure homogenize, process, solution is removed organic solvent micromolecule by dialysis or ultrafiltration or post partition method and is obtained the nano-micelle that particle diameter is 10～1000nm, the nano-micelle obtaining can directly be applied, and also can recycle by lyophilization or dry the making after solid preparation redissolves of spraying.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310742883.9A CN103705940B (en) | 2013-12-30 | 2013-12-30 | The preparation of a kind of natural activity medicine-polysaccharide targeting complex and antineoplastic application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310742883.9A CN103705940B (en) | 2013-12-30 | 2013-12-30 | The preparation of a kind of natural activity medicine-polysaccharide targeting complex and antineoplastic application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103705940A true CN103705940A (en) | 2014-04-09 |
| CN103705940B CN103705940B (en) | 2016-03-02 |
Family
ID=50399522
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310742883.9A Active CN103705940B (en) | 2013-12-30 | 2013-12-30 | The preparation of a kind of natural activity medicine-polysaccharide targeting complex and antineoplastic application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103705940B (en) |
Cited By (23)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104398493A (en) * | 2014-12-08 | 2015-03-11 | 中国人民解放军第四军医大学 | Tumor active-targeting nano drug delivery system capable of reversing drug-resistance |
| CN106267226A (en) * | 2015-06-09 | 2017-01-04 | 北京和理咨询有限公司 | Treat osteosarcomatous triptolide derivant and preparation method thereof |
| CN106860874A (en) * | 2017-01-23 | 2017-06-20 | 福建中医药大学 | Carboxymethyl chitosan Rhein conjugate and its synthesis technique |
| CN107050465A (en) * | 2017-01-17 | 2017-08-18 | 中国药科大学 | A kind of feature Nano medication of curcumin and its application for brain diseases diagnosis and treatment |
| CN107400180A (en) * | 2017-07-27 | 2017-11-28 | 大连民族大学 | Redox responds the preparation method and purposes of chitosan liposome |
| CN108079307A (en) * | 2018-02-08 | 2018-05-29 | 中国药科大学 | A kind of tri compound nanometer system and its application based on methoxy polyethylene glycol-polylactic acid |
| CN108409878A (en) * | 2018-01-31 | 2018-08-17 | 湖北民族学院 | Cortex magnoliae officinalis polysaccharide and its preparation and use |
| CN108904447A (en) * | 2018-08-15 | 2018-11-30 | 烟台大学 | A kind of liver tumour targeted carrier material, micellar preparation and preparation method thereof |
| CN109464675A (en) * | 2018-11-29 | 2019-03-15 | 河南中医药大学 | A kind of preparation method and application of triptolide-carboxyl chitosan coupling drug |
| CN109481694A (en) * | 2018-12-20 | 2019-03-19 | 药大制药有限公司 | A kind of antiviral conjugate of Ribavirin-resveratrol, preparation method and application |
| CN109481400A (en) * | 2018-12-21 | 2019-03-19 | 潍坊医学院 | A kind of Liver targeting adriamycin/Bcl-2 siRNA carries nano-micelle and its preparation method and application altogether |
| CN109678986A (en) * | 2018-11-30 | 2019-04-26 | 河南大学 | A kind of rutin chelate and its preparation method and application |
| CN110201216A (en) * | 2019-07-24 | 2019-09-06 | 中国人民解放军陆军军医大学第二附属医院 | One kind is for through and through wound rapid hemostatic material and preparation method thereof |
| WO2019189419A1 (en) * | 2018-03-28 | 2019-10-03 | 持田製薬株式会社 | Anti-cancer agent-bonded alginic acid derivative |
| CN110859825A (en) * | 2019-11-26 | 2020-03-06 | 武汉理工大学 | Preparation method of targeted drug delivery nano-delivery system |
| CN111388449A (en) * | 2020-04-08 | 2020-07-10 | 华侨大学 | Polyornithine/carboxymethyl lentinan layer-by-layer self-assembly drug carrier and preparation method thereof |
| CN111870580A (en) * | 2020-08-04 | 2020-11-03 | 温州医科大学附属第二医院、温州医科大学附属育英儿童医院 | Curcumin-containing medicament and application thereof in targeted therapy of colitis |
| CN113061180A (en) * | 2021-04-02 | 2021-07-02 | 山东大学 | Conjugate of ES2 peptide without anticoagulation activity heparin modification and combination with paclitaxel, preparation method and application thereof |
| CN114522171A (en) * | 2021-12-06 | 2022-05-24 | 福建医科大学孟超肝胆医院(福州市传染病医院) | Lentinan ursolic acid self-assembly nanoparticles and preparation method and application thereof |
| CN115317451A (en) * | 2022-07-29 | 2022-11-11 | 河南中医药大学 | Cel-CSO/Taxol NPs nano-medicine and application thereof |
| CN115554240A (en) * | 2021-07-02 | 2023-01-03 | 福建中医药大学 | Tripterine/glycyrrhetinic acid-carboxymethyl chitosan-ketothiol-rhein micelle |
| CN115558039A (en) * | 2021-07-02 | 2023-01-03 | 福建中医药大学 | Glycyrrhetinic acid-carboxymethyl chitosan-keto thiol-rhein conjugate, and preparation method and application thereof |
| CN116496430A (en) * | 2023-06-26 | 2023-07-28 | 中国科学院烟台海岸带研究所 | Amphiphilic chitosan cystamine grafted derivative and preparation and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101791411A (en) * | 2010-01-25 | 2010-08-04 | 中国药科大学 | Preparation and application of amphiphilic polysaccharide conjugate and medicinal compositions thereof |
| CN102988999A (en) * | 2012-05-09 | 2013-03-27 | 中国药科大学 | Curcumin-polysaccharide conjugate as well as preparation method and application thereof |
-
2013
- 2013-12-30 CN CN201310742883.9A patent/CN103705940B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101791411A (en) * | 2010-01-25 | 2010-08-04 | 中国药科大学 | Preparation and application of amphiphilic polysaccharide conjugate and medicinal compositions thereof |
| CN102988999A (en) * | 2012-05-09 | 2013-03-27 | 中国药科大学 | Curcumin-polysaccharide conjugate as well as preparation method and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 邓启盼: "纳米脂质体应用于肿瘤研究的进展", 《纳米脂质体应用于肿瘤研究的进展》, vol. 30, no. 6, 31 December 2010 (2010-12-31), pages 489 - 494 * |
Cited By (33)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104398493B (en) * | 2014-12-08 | 2017-06-30 | 中国人民解放军第四军医大学 | A kind of tumor-targeting nanoscale medicine delivery system of reversible tumor drug resistance |
| CN104398493A (en) * | 2014-12-08 | 2015-03-11 | 中国人民解放军第四军医大学 | Tumor active-targeting nano drug delivery system capable of reversing drug-resistance |
| CN106267226A (en) * | 2015-06-09 | 2017-01-04 | 北京和理咨询有限公司 | Treat osteosarcomatous triptolide derivant and preparation method thereof |
| CN107050465A (en) * | 2017-01-17 | 2017-08-18 | 中国药科大学 | A kind of feature Nano medication of curcumin and its application for brain diseases diagnosis and treatment |
| CN106860874A (en) * | 2017-01-23 | 2017-06-20 | 福建中医药大学 | Carboxymethyl chitosan Rhein conjugate and its synthesis technique |
| CN107400180A (en) * | 2017-07-27 | 2017-11-28 | 大连民族大学 | Redox responds the preparation method and purposes of chitosan liposome |
| CN108409878A (en) * | 2018-01-31 | 2018-08-17 | 湖北民族学院 | Cortex magnoliae officinalis polysaccharide and its preparation and use |
| CN108079307A (en) * | 2018-02-08 | 2018-05-29 | 中国药科大学 | A kind of tri compound nanometer system and its application based on methoxy polyethylene glycol-polylactic acid |
| WO2019189419A1 (en) * | 2018-03-28 | 2019-10-03 | 持田製薬株式会社 | Anti-cancer agent-bonded alginic acid derivative |
| CN108904447A (en) * | 2018-08-15 | 2018-11-30 | 烟台大学 | A kind of liver tumour targeted carrier material, micellar preparation and preparation method thereof |
| CN108904447B (en) * | 2018-08-15 | 2020-10-30 | 烟台大学 | Liver tumor targeting carrier material, micelle preparation and preparation method thereof |
| CN109464675A (en) * | 2018-11-29 | 2019-03-15 | 河南中医药大学 | A kind of preparation method and application of triptolide-carboxyl chitosan coupling drug |
| CN109464675B (en) * | 2018-11-29 | 2021-10-29 | 河南中医药大学 | Preparation method and application of triptolide-carboxylated chitosan coupling drug |
| CN109678986B (en) * | 2018-11-30 | 2020-12-22 | 河南大学 | Rutin chelate and preparation method and application thereof |
| CN109678986A (en) * | 2018-11-30 | 2019-04-26 | 河南大学 | A kind of rutin chelate and its preparation method and application |
| CN109481694A (en) * | 2018-12-20 | 2019-03-19 | 药大制药有限公司 | A kind of antiviral conjugate of Ribavirin-resveratrol, preparation method and application |
| CN109481400A (en) * | 2018-12-21 | 2019-03-19 | 潍坊医学院 | A kind of Liver targeting adriamycin/Bcl-2 siRNA carries nano-micelle and its preparation method and application altogether |
| CN109481400B (en) * | 2018-12-21 | 2021-03-16 | 潍坊医学院 | Liver-targeted adriamycin/Bcl-2 siRNA co-loaded nano micelle and preparation method and application thereof |
| CN110201216A (en) * | 2019-07-24 | 2019-09-06 | 中国人民解放军陆军军医大学第二附属医院 | One kind is for through and through wound rapid hemostatic material and preparation method thereof |
| CN110859825A (en) * | 2019-11-26 | 2020-03-06 | 武汉理工大学 | Preparation method of targeted drug delivery nano-delivery system |
| CN110859825B (en) * | 2019-11-26 | 2021-12-07 | 武汉理工大学 | Preparation method of targeted drug delivery nano-delivery system |
| CN111388449A (en) * | 2020-04-08 | 2020-07-10 | 华侨大学 | Polyornithine/carboxymethyl lentinan layer-by-layer self-assembly drug carrier and preparation method thereof |
| CN111870580A (en) * | 2020-08-04 | 2020-11-03 | 温州医科大学附属第二医院、温州医科大学附属育英儿童医院 | Curcumin-containing medicament and application thereof in targeted therapy of colitis |
| CN113061180A (en) * | 2021-04-02 | 2021-07-02 | 山东大学 | Conjugate of ES2 peptide without anticoagulation activity heparin modification and combination with paclitaxel, preparation method and application thereof |
| CN115554240A (en) * | 2021-07-02 | 2023-01-03 | 福建中医药大学 | Tripterine/glycyrrhetinic acid-carboxymethyl chitosan-ketothiol-rhein micelle |
| CN115558039A (en) * | 2021-07-02 | 2023-01-03 | 福建中医药大学 | Glycyrrhetinic acid-carboxymethyl chitosan-keto thiol-rhein conjugate, and preparation method and application thereof |
| CN115558039B (en) * | 2021-07-02 | 2023-05-12 | 福建中医药大学 | Glycyrrhetinic acid-carboxymethyl chitosan-ketal-rhetinic acid conjugate, preparation method and application thereof |
| CN115554240B (en) * | 2021-07-02 | 2023-09-15 | 福建中医药大学 | Tripterine/glycyrrhetinic acid-carboxymethyl chitosan-ketal-rhein micelle |
| CN114522171A (en) * | 2021-12-06 | 2022-05-24 | 福建医科大学孟超肝胆医院(福州市传染病医院) | Lentinan ursolic acid self-assembly nanoparticles and preparation method and application thereof |
| CN114522171B (en) * | 2021-12-06 | 2024-02-27 | 福建医科大学孟超肝胆医院(福州市传染病医院) | Lentinan ursolic acid self-assembled nano-particles and preparation method and application thereof |
| CN115317451A (en) * | 2022-07-29 | 2022-11-11 | 河南中医药大学 | Cel-CSO/Taxol NPs nano-medicine and application thereof |
| CN116496430A (en) * | 2023-06-26 | 2023-07-28 | 中国科学院烟台海岸带研究所 | Amphiphilic chitosan cystamine grafted derivative and preparation and application thereof |
| CN116496430B (en) * | 2023-06-26 | 2023-08-25 | 中国科学院烟台海岸带研究所 | Amphiphilic chitosan cystamine grafted derivative and preparation and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103705940B (en) | 2016-03-02 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103705940B (en) | The preparation of a kind of natural activity medicine-polysaccharide targeting complex and antineoplastic application thereof | |
| CN103705939B (en) | The amphipathic preparation of ursolic acid-polysaccharide conjugate and the application in oncotherapy thereof | |
| CN101791411B (en) | Preparation and application of amphiphilic polysaccharide conjugate and medicinal compositions thereof | |
| CN102988999B (en) | Curcumin-polysaccharide conjugate as well as preparation method and application thereof | |
| CN103705534A (en) | Preparation of natural active substance constructed polymer composite medicine and application thereof in inhibiting angiogenesis | |
| CN102125547B (en) | Pharmaceutical composition containing gambogic acid medicament and preparation method thereof | |
| CN103301472A (en) | Amphiphilic polysaccharide-anti-tumor medicament conjugate capable of releasing medicines specifically at lesion site of living body, as well as preparation method and application of medicinal composition of amphiphilic polysaccharide-anti-tumor medicament conjugate | |
| Fan et al. | Plasma membrane targeted photodynamic O2 economizer for hypoxic tumor therapy | |
| CN106267229B (en) | A kind of Liver targeting carries the structure and preparation method thereof of platinum nano-prodrug | |
| CN109875964B (en) | Preparation and application of adriamycin carrier-free nano-drug | |
| CN110408047B (en) | Nano coordination polymer and preparation method and application thereof | |
| Liu et al. | Polymeric prodrug of bufalin for increasing solubility and stability: Synthesis and anticancer study in vitro and in vivo | |
| CN103705943A (en) | Preparation method and application of reduction-response-type pegylation (PEG) nanomedicine composition | |
| Yi et al. | Synthesis, characterization, and formulation of poly-puerarin as a biodegradable and biosafe drug delivery platform for anti-cancer therapy | |
| CN104940945A (en) | Hollow mesoporous copper sulfide compound modified by hyaluronic acid and preparation method and application thereof | |
| CN105273205A (en) | Block polymer with benzeneboronic acid ester as connecting unit, synthesis method and application thereof | |
| Wang et al. | Glucose oxidase-amplified CO generation for synergistic anticancer therapy via manganese carbonyl-caged MOFs | |
| CN111053911A (en) | Reduction response type cross-linking agent and preparation and application of cross-linked hydroxyl drug molecule thereof | |
| CN106420664A (en) | Application of adopting aspirin conjugate with antitumor activity as drug carrier or molecular probe carrier | |
| Liu et al. | Self-delivery nanomedicine for vascular disruption-supplemented chemo-photodynamic tumor therapy | |
| CN109846857B (en) | Preparation method and application of active natural supramolecular photosensitizer | |
| CN111358948A (en) | Camptothecin-berberine/indocyanine green nano-drug, preparation method and application | |
| CN107375199A (en) | A kind of nanogel delivery system for polymerizeing chloroquine and preparation method thereof | |
| CN108888774B (en) | Tripterine-dendrimer conjugate as well as preparation method and application thereof | |
| CN104208704A (en) | Preparation method for pH-sensitive carbon nano tube-targeted drug delivery system |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20190628 Address after: Room 111 and 113, Building 9, Weidi Road, Qixia District, Nanjing City, Jiangsu Province Patentee after: Nanjing Kening Testing Technology Co.,Ltd. Address before: 211198 No. 639 Longmian Avenue, Jiangning District, Nanjing City, Jiangsu Province Patentee before: China Pharmaceutical University |
|
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20230927 Address after: Room 903, 905, and 907, Building D6, Jiangsu Life Science Park, No. 9 Weidi Road, Xianlin Street, Qixia District, Nanjing City, Jiangsu Province, 210000 Patentee after: NANJING ZEHENG PHARMACEUTICAL SCIENCE & TECHNOLOGY Co.,Ltd. Address before: Room 111 and 113, Building 9, Weidi Road, Qixia District, Nanjing City, Jiangsu Province Patentee before: Nanjing Kening Testing Technology Co.,Ltd. |
|
| TR01 | Transfer of patent right |