CN107875399A - A kind of Telmisartan modified chitosan oligosaccharide fatty acid nanoparticle and preparation and application - Google Patents
A kind of Telmisartan modified chitosan oligosaccharide fatty acid nanoparticle and preparation and application Download PDFInfo
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- CN107875399A CN107875399A CN201711019493.3A CN201711019493A CN107875399A CN 107875399 A CN107875399 A CN 107875399A CN 201711019493 A CN201711019493 A CN 201711019493A CN 107875399 A CN107875399 A CN 107875399A
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Abstract
The present invention provides Telmisartan modified chitosan oligosaccharide fatty acid, is polymer substance, the amino wherein on chitosan oligosaccharide is substituted by aliphatic acid and Telmisartan molecule.Wherein chitosan oligosaccharide fatty acid molecular weight is 1~20kDa, and deacetylation is 70%~100%, and the carbon chain lengths of aliphatic acid are C12 C22, and amino group substitution degree is 1%~50%.The present invention is in Stability Analysis of Structures, passive target function of tumor is strong, on the high chitosan oligosaccharide fatty acid architecture basics of internal stability, the Telmisartan of chemical modification small-molecular-weight again, not only increase the stability of medicine in vivo, also by being interacted with the blood angiotonin II receptor I of the high expression of tumor cell surface, possess active targeting tumour cell ability, antineoplastic can be improved the oncotherapy the effect of.Its representational general structure is：
Description
Technical field
The invention belongs to art of pharmacy, is related to a kind of Telmisartan modified chitosan oligosaccharide fatty acid nanoparticle and preparation method,
And the application of synthesized Telmisartan modified chitosan oligosaccharide fatty acid nanoparticle.
Background technology
Tumour is still to threaten the most important disease of human life quality, although the treatment method such as chemotherapy, radiotherapy can be with
Extend the survival period of patient, but distribution lacks specificity in small molecule, anti-tumor drug body, easily damages human normal organ,
Such as existing doxorubicin hydrochloride injection, the side effects such as serious myocardial degenerative disease, bone marrow suppression are easily produced, are limited
Clinical practice.
The administration nano-drug administration system of free drug is encapsulated, can be by strengthening infiltration and interception effect, Passive diffusion to tumour
Tissue, improves antitumor drug effect.Although Nano medication improves the life cycle of patient in terms for the treatment of, clinical treatment outcome is simultaneously
Without optimistic as preclinical laboratory result.Therefore, researcher utilizes normal cell and neoplastic cell receptor expression
Difference, by realizing neoplastic disease in administration nano-drug administration system surface modification target head such as monoclonal antibody, polypeptide, accounting, aptamers etc.
More assemble at stove position.
Chitosan is a kind of cationic polymer being made up of Glucosamine, by alkali or enzyme chitin can be made to take off second
Acyl group and obtain.This Natural polycations natural macromolecular material has preferable biocompatibility, biodegradability, peace
Quan Xing.Cell to liposoluble substance to having higher intake ability, but chitosan oligosaccharide is due to a lack of lipophilicity, it is not easy to through cell
Film.With lipophilicity aliphatic acid by chemical grafting, the amphipathy macromolecule being prepared can pass through chitosan oligosaccharide in an aqueous medium
The mode of self aggregation forms micella.The secondary core that the nanoparticle is formed due to the aliphatic acid of modification in micellar surface, can be swollen
Oncocyte quickly absorbs.After the micella contains antineoplastic by way of physically encapsulation, it is remarkably improved original antitumor
The therapeutic activity of medicine and the function of reversing tumor cellular drug resistance.
Existing research has confirmed that blood angiotonin II receptor I is the g protein coupled receptor family expressed on cell membrane,
And high expression, such as breast cancer, cancer of pancreas, oophoroma etc. on kinds of tumor cells film.Therefore blood angiotonin II receptor I can
With as the potential acceptor for realizing that tumor microenvironment targets.
Blood angiotonin II receptor I inhibitor such as Telmisartan is used for protecting that hypertensive patient's is dirty by FDA approvals
Device.Have scholar also verified Telmisartan be in ARBs class medicines with AT1R affinity is most strong.
By the part amino grafting in carboxylic Telmisartan and chitosan oligosaccharide fatty acid structure, the product being prepared exists
The micella that active targeting acts on can be formed in water by self aggregation.The hydrophobic cores of the micella can be with solubilisation of hydrophobic medicine
Molecule.Due to the Telmisartan of surface modification, drug delivery system can improve the ingestion of medicines of cell, increase antineoplaston
Drug effect.The application of the active targeting carrier material, intake and increasing of the tumour cell to antineoplastic can be increased substantially
Add the aggregation of tumor locus medicine, the final treatment drug effect for improving patient.
The content of the invention
It is an object of the present invention to provide a kind of Telmisartan modified chitosan oligosaccharide fatty acid nanoparticle, its is representative
General structure be：
Wherein：
The sugared monocyclic percentage that a is not substituted for amino in Telmisartan modified chitosan oligosaccharide fatty acid nanoparticle, scope
For 40%~90%,
B is the sugared monocyclic percentage substituted in Telmisartan modified chitosan oligosaccharide fatty acid nanoparticle by aliphatic acid, and scope is
1%~50%,
C is the sugared monocyclic percentage that is substituted by Telmisartan in Telmisartan modified chitosan oligosaccharide fatty acid nanoparticle, scope
For 0.3%~2.0%.
Second object of the present invention is to provide the preparation method of Telmisartan modified chitosan oligosaccharide fatty acid, by with lower section
Case is realized：
1. prepared by reaction solution：15mmol~30mmol Telmisartan is weighed, is dissolved in 1.5mL DMFs,
Add 150mmol n-hydroxysuccinimides and 150mmol 1- (3- dimethylamino-propyls) -3- ethyl carbodiimide hydrochlorides
Salt, 60 DEG C activate 1 hour；
2. weighing chitosan oligosaccharide fatty acid 2mmol, it is dissolved in 4mL water, Probe Ultrasonic Searching 30 times, adds above-mentioned reaction solution, and mend
Add 4.0mL DMFs, 60 DEG C are reacted 12 hours；Reaction solution is put in bag filter, dialysed in the acid solutions of pH 1.0
2 days, distilled water continued dialysis 2 days, freeze-drying, obtains purpose product Telmisartan modified chitosan oligosaccharide fatty acid.
Chitosan oligosaccharide fatty acid used in the present invention has been national inventing patent " fluorescence labeling hydrophobic modified chitosan polymerization
Thing and the preparation method and application " (patent No.：ZL2005100507981) and " surface-modified hydrophobically modified chitin polymer is given
Medicine micelle and preparation method thereof " (patent No.：ZL200610051601.0) covered.Chitosan oligosaccharide fatty acid amino group substitution degree is
1%~50%；Wherein 1~200kDa of molecular weight of chitosan oligosaccharide；The carbon chain lengths of aliphatic acid are：C12-C22；The de- second of chitosan oligosaccharide
Acyl degree is 70%-100%.
Third object of the present invention is to provide Telmisartan modified chitosan oligosaccharide fatty acid and is preparing anti-tumor immunotherapy
Application in medicine, the specifically application in the anti-tumor immunotherapy medicine for preparing targeting breast cancer tumor cells.Result of study
Show that Telmisartan modified chitosan oligosaccharide fatty acid can be with active targeting breast cancer tumor cells.
Telmisartan modified chitosan oligosaccharide fatty acid provided by the invention, is polymer substance, the amino wherein on chitosan oligosaccharide
Substituted by aliphatic acid and Telmisartan molecule.Wherein chitosan oligosaccharide fatty acid molecular weight is 1~20kDa, deacetylation is 70%~
100%, the carbon chain lengths of aliphatic acid are C12-C22, and amino group substitution degree is 1%~50%.The present invention is in Stability Analysis of Structures, by moving-target
Strong to function of tumor, on the high chitosan oligosaccharide fatty acid architecture basics of internal stability, then chemical modification small-molecular-weight replaces meter Sha
It is smooth, the stability of medicine in vivo is not only increased, also passes through the blood angiotonin II receptor I with the high expression of tumor cell surface
Interaction, possesses active targeting tumour cell ability, can improve antineoplastic the oncotherapy the effect of.The present invention is excellent
Gesture is：On the basis of the research work of early stage, the abundant amino to dissociate on chitosan oligosaccharide fatty acid and Telmisartan molecule are utilized
The upper carboxyl with reactivity reacts under catalyst action, and synthesis obtains Telmisartan modified chitosan oligosaccharide fatty acid delivering system
System, improves stability inside medicine, can improve intake and antitumor drug effect of the tumour cell to medicine.
Brief description of the drawings
Fig. 1 is the nuclear magnetic resonance map of Telmisartan, chitosan oligosaccharide fatty acid and Telmisartan modified chitosan oligosaccharide fatty acid.
Fig. 2 is that adriamycin, chitosan oligosaccharide fatty acid carrier micelle and Telmisartan modified chitosan oligosaccharide fatty acid carrier micelle exist
The release profiles of adriamycin in different pH dissolution mediums.
Fig. 3 is the carrier of the Telmisartan modified chitosan oligosaccharide fatty acid of flow cytomery difference bonding ratio in mammary gland
Result is absorbed on cancer cell.
Embodiment
The present invention is further described by embodiment and accompanying drawing.
Embodiment 1
According to national inventing patent " fluorescence labeling hydrophobic modified chitosan polymer and preparation method and application " (patent
Number：) and " surface-modified hydrophobically modified chitin polymer administration micelle and preparation method thereof " (patent ZL2005100507981
Number：ZL200610051601.0), it is synthetically prepared to obtain chitosan oligosaccharide fatty acid.
The stearic amino group substitution degree of chitosan oligosaccharide is determined using TNB method, takes the shell of 1~10mg different weights
Oligosaccharides is dissolved in 2mL distilled water respectively, adds 4% sodium acid carbonate 2mL and 0.1% TNB 2mL, 2 are incubated at 37 DEG C
Hour, 2mol/L hydrochloric acid 2mL are added, are shaken up, absorbance is determined at 344nm, standard curve is prepared, takes above-mentioned chitosan oligosaccharide fatty acid
4mg, it is dissolved in 2mL distilled water, is operated with method, it is 6.71% to calculate chitosan oligosaccharide-stearic amino group substitution degree by standard curve；
The preparation of Telmisartan modified chitosan oligosaccharide fatty acid：
Telmisartan 7.71mg is taken, is dissolved in 1.5mL DMFs, adds 17.25mg N- hydroxysuccinimidyl acyls
Imines and 28.65mg 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides, 60 DEG C activate 1 hour；
Chitosan oligosaccharide fatty acid 40mg is weighed, is dissolved in 4mL water, Probe Ultrasonic Searching 30 times, adds above-mentioned reaction solution whole, and mend
Add 4.0mL DMFs, 60 DEG C are reacted 12 hours.Reaction solution is put in bag filter, dialysed in the acid solutions of pH 1.0
2 days, distilled water continued dialysis 2 days, freeze-drying, obtains Telmisartan modified chitosan oligosaccharide fatty acid.
4mg chitosan oligosaccharide fatty acids and Telmisartan modified chitosan oligosaccharide fatty acid are weighed respectively, add 2mL water, Probe Ultrasonic Searching
30 times, the μ l of 2mg/mL fluorescein isothiocynates 200 are taken respectively, and room temperature lucifuge stirs 12 hours.Reaction solution is put in bag filter, thoroughly
Analysis 1 day, centrifugation, obtain the chitosan oligosaccharide fatty acid and Telmisartan modified chitosan oligosaccharide fatty acid of marked by fluorescein isothiocyanate.
In 24 porocyte culture plates, 1mL is added per hole and contains 1 × 105Individual MCF-7 cells, put 37 DEG C, 5%CO2Incubator is trained
Support 12 hours, after cell is completely adherent, 100 μ l Telmisartan modified chitosan oligosaccharide fatty acids are added per hole.It is small to continue incubation 12
When, nutrient solution is discarded, is cleaned three times with isotonic phosphate buffer, cell dissociation is got off with 0.25% pancreatin, centrifugation obtains
Cell precipitation is obtained, cell precipitation, the fluorescent value of flow cytomery fluorescein isothiocynate are resuspended with 0.5mL buffer solutions.Inspection
Survey result and show that chitosan oligosaccharide fatty acid and Telmisartan modified chitosan oligosaccharide fatty acid are respectively 12.8 and 22.1.
Embodiment 2
According to national inventing patent " fluorescence labeling hydrophobic modified chitosan polymer and preparation method and application " (patent
Number：) and " surface-modified hydrophobically modified chitin polymer administration micelle and preparation method thereof " (patent ZL2005100507981
Number：ZL200610051601.0), it is synthetically prepared to obtain chitosan oligosaccharide fatty acid.
The stearic amino group substitution degree of chitosan oligosaccharide is determined using TNB method, takes the shell of 1~10mg different weights
Oligosaccharides is dissolved in 2mL distilled water respectively, adds 4% sodium acid carbonate 2mL and 0.1% TNB 2mL, 2 are incubated at 37 DEG C
Hour, 2mol/L hydrochloric acid 2mL are added, are shaken up, absorbance is determined at 344nm, standard curve is prepared, takes above-mentioned chitosan oligosaccharide fatty acid
4mg is dissolved in 2mL distilled water, is operated with method, and it is 6.71% to calculate chitosan oligosaccharide-stearic amino group substitution degree by standard curve；
The preparation of Telmisartan modified chitosan oligosaccharide fatty acid：
Telmisartan 11.56mg is taken, is dissolved in 1.5mL DMFs, adds 17.25mg N- hydroxysuccinimidyl acyls
Imines and 28.65mg 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides, 60 DEG C activate 1 hour；
Chitosan oligosaccharide fatty acid 40mg is weighed, is dissolved in 4mL water, Probe Ultrasonic Searching 30 times, adds above-mentioned reaction solution whole, and mend
Add 4.0mL DMFs, 60 DEG C are reacted 12 hours.Reaction solution is put in bag filter, dialysed in the acid solutions of pH 1.0
2 days, distilled water continued dialysis 2 days, freeze-drying, obtains Telmisartan modified chitosan oligosaccharide fatty acid product.
4mg chitosan oligosaccharide fatty acids and Telmisartan modified chitosan oligosaccharide fatty acid are weighed respectively, add 2mL water, Probe Ultrasonic Searching
30 times, the μ l of 2mg/mL fluorescein isothiocynates 200 are taken respectively, and room temperature lucifuge stirs 12 hours.Reaction solution is put in bag filter, thoroughly
Analysis 1 day, centrifugation, obtain the chitosan oligosaccharide fatty acid and Telmisartan modified chitosan oligosaccharide fatty acid of marked by fluorescein isothiocyanate.
In 24 porocyte culture plates, 1mL is added per hole and contains 1 × 105Individual MCF-7 cells, put 37 DEG C, 5%CO2Incubator is trained
Support 12 hours, after cell is completely adherent, 100 μ l Telmisartan modified chitosan oligosaccharide fatty acids are added per hole.It is small to continue incubation 12
When, nutrient solution is discarded, is cleaned three times with isotonic phosphate buffer, cell dissociation is got off with 0.25% pancreatin, centrifugation obtains
Cell precipitation is obtained, cell precipitation, the fluorescent value of flow cytomery fluorescein isothiocynate are resuspended with 0.5mL buffer solutions.Inspection
Survey result and show that chitosan oligosaccharide fatty acid and Telmisartan modified chitosan oligosaccharide fatty acid are respectively 12.8 and 35.8.
Embodiment 3
According to national inventing patent " fluorescence labeling hydrophobic modified chitosan polymer and preparation method and application " (patent
Number：) and " surface-modified hydrophobically modified chitin polymer administration micelle and preparation method thereof " (patent ZL2005100507981
Number：ZL200610051601.0), it is synthetically prepared to obtain chitosan oligosaccharide fatty acid.
The stearic amino group substitution degree of chitosan oligosaccharide is determined using TNB method, takes the shell of 1~10mg different weights
Oligosaccharides is dissolved in 2mL distilled water respectively, adds 4% sodium acid carbonate 2mL and 0.1% TNB 2mL, 2 are incubated at 37 DEG C
Hour, 2mol/L hydrochloric acid 2mL are added, are shaken up, absorbance is determined at 34nm, standard curve is prepared, takes above-mentioned chitosan oligosaccharide fatty acid
4mg is dissolved in 2mL distilled water, is operated with method, and it is 6.71% to calculate chitosan oligosaccharide-stearic amino group substitution degree by standard curve.
The preparation of Telmisartan modified chitosan oligosaccharide fatty acid：
Telmisartan 15.42mg is taken, is dissolved in 1.5mL DMFs, adds 17.25mg N- hydroxysuccinimidyl acyls
Imines and 28.65mg 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides, 60 DEG C activate 1 hour；
Chitosan oligosaccharide fatty acid 40mg is weighed, is dissolved in 4mL water, Probe Ultrasonic Searching 30 times, adds above-mentioned reaction solution whole, and mend
Add 4.0mL DMFs, 60 DEG C are reacted 12 hours.Reaction solution is put in bag filter, dialysed in the acid solutions of pH 1.0
2 days, distilled water continued dialysis 2 days, freeze-drying, obtains Telmisartan modified chitosan oligosaccharide fatty acid product.
Nuclear magnetic resonance spectrometry measure Telmisartan, chitosan oligosaccharide stearic acid and Telmisartan modified chitosan oligosaccharide fatty acid.Claim
Chitosan oligosaccharide fatty acid and each 5mg of Telmisartan modified chitosan oligosaccharide fatty acid are taken, uses 0.5mLD respectively2O dissolves, and separately takes Telmisartan
5mg, 0.5mLDMSO-d6 is dissolved in, uses nuclear magnetic resonance1H-NMR is determined, and as a result referring to Fig. 1, A Telmisartans in figure, B is chitosan oligosaccharide
Aliphatic acid, C are Telmisartan modified chitosan oligosaccharide fatty acid.
Using the critical micelle concentration of pyrene fluorescence spectrometry chitosan oligosaccharide fatty acid and Telmisartan modified chitosan oligosaccharide fatty acid.
Accurately weighed pyrene 12mg, puts 100mL volumetric flasks, adds acetone solution and constant volume.Above-mentioned pyrene solution 1mL is pipetted, puts 100mL volumetric flasks
Middle dilution and constant volume.Pipette the pyrene solution 0.5mL after dilution to put respectively in 10mL teat glasses, 50 DEG C are flung to acetone.Respectively plus
Enter the chitosan oligosaccharide fatty acid and Telmisartan modified chitosan oligosaccharide fatty acid solution 5mL of various concentrations, control pyrene final concentration of 7 ×
10-7Mol/l, room-temperature water bath ultrasound 30min.Scan the excitation spectrum and emission spectrum (EX=337nm, EM of pyrene：I1=374nm,
I3=384nm, slit=2.5nm and 10nm), fluorescence intensity is determined, and critical micelle concentration is calculated, respectively 48.3 μ g/mL
With 16.1 μ g/mL.
Precision weighs chitosan oligosaccharide fatty acid and Telmisartan modified chitosan oligosaccharide fatty acid 4mg respectively, adds 2mL water, probe
Ultrasound 30 times, is separately added into the μ l of 2mg/mL adriamycins dimethyl sulfoxide solution 200.After magnetic agitation 2 hours, bag filter dialysis is placed in
2 days, obtain chitosan oligosaccharide fatty acid carrier micelle and Telmisartan modified chitosan oligosaccharide fatty acid carrier micelle.
Utilize fluorescence spectrophotometry chitosan oligosaccharide fatty acid carrier micelle and Telmisartan modified chitosan oligosaccharide fatty acid
The content of adriamycin in carrier micelle, adriamycin are dissolved in dimethyl sulfoxide (DMSO), and dimethyl sulfoxide dilutes the solution for being made into various concentrations, system
Standby standard curve.A certain amount of chitosan oligosaccharide fatty acid carrier micelle and Telmisartan modified chitosan oligosaccharide fatty acid carrier micelle is dissolved in two
In first sulfoxide, fluorescent value is determined, it is respectively 8.0% He to calculate doxorubicin content (percentage by weight) in carrier by standard curve
7.5%.
Weigh chitosan oligosaccharide fatty acid, Telmisartan modified chitosan oligosaccharide fatty acid each 10mg is accurately weighed, is dissolved in 10mL steamings
Distilled water, obtain 1mg/mL solution.Probe Ultrasonic Searching 30 times (400w, work 2s, interval 3s).Particle size is analyzed with surface potential
It is 62.3nm and 76.3nm that instrument, which determines chitosan oligosaccharide fatty acid micella and the particle diameter of Telmisartan modified chitosan oligosaccharide fatty acid micella,
Zeta potential is 16.7mV and 35.1mV.
Chitosan oligosaccharide fatty acid carrier micelle and each 10mg of Telmisartan modified chitosan oligosaccharide fatty acid carrier micelle separately are weighed, essence
It is close weighed, 10mL distilled water is dissolved in, obtains 1mg/mL solution.Particle size and surface potential analysis-e/or determining chitosan oligosaccharide fat
The particle diameter that fat acid carrier micelle and Telmisartan modified chitosan oligosaccharide fatty acid carry is 34.6nm and 30.3nm.
Chitosan oligosaccharide fatty acid carrier micelle and the investigation of Telmisartan modified chitosan oligosaccharide fatty acid release in vitro behavior：
Doxorubicin concentration is taken respectively as 30 μ g/mL chitosan oligosaccharide fatty acids carrier micelles and Telmisartan modified chitosan oligosaccharide fat
Sour carrier micelle 1mL, it is put into bag filter (MWCO=7000), the phosphate buffer that bag filter is put into 20mL differences pH (divides
Wei pH 5.0 and pH 7.4) in, vibrated in 37 DEG C of shaking table.Sampled in different time points, whole releases are discarded after sampling
Medium, add fresh medium 20mL, continuous sampling 72 hours.Drug concentration (EX=in fluorescence spectrophotometry sample
505nm, EM=565nm, slit=5nm).Release profiles are shown in Fig. 2.
The antitumor drug effect of chitosan oligosaccharide fatty acid carrier micelle and Telmisartan modified chitosan oligosaccharide fatty acid carrier micelle：
The present invention is used with inhibition rate of tumor cell, evaluation Telmisartan modified chitosan oligosaccharide fatty acid cell viability assays
Tetrazolium salt colorimetric assay determines.Using breast cancer cell MCF-7 cells as model, in 96 porocyte culture plates, 200 μ l are added per hole
Containing 2 × 103The nutrient solution of individual MCF-7 cells, put 37 DEG C, 5%CO2Incubator culture 12 hours, after cell is completely adherent, cell
Doxorubicin solution, chitosan oligosaccharide fatty acid carrier micelle and Telmisartan the modified chitosan oligosaccharide fat of various concentrations are separately added into hole
Sour carrier micelle, using undressed blanc cell as control, multiple holes are set per hole；After being incubated 48 hours, 5mg/mL is added per hole
The μ l of Thiazolyl blue solution 20, continue abandoning supernatant after being incubated 4 hours, the μ l of dimethyl sulfoxide (DMSO) 200 are added per hole, use enzyme detector
Absorbance at 570nm is determined, cell survival rate is calculated as follows：
Cell survival rate (%)=experimental group absorbance/control group absorbance × 100%
It is computed, adriamycin, chitosan oligosaccharide fatty acid carrier micelle and Telmisartan modified chitosan oligosaccharide fatty acid carrier micelle
IC50 to MCF-7 cells is respectively 1.80 μ g/mL, 4.39 μ g/mL and 2.49 μ g/mL.
4mg chitosan oligosaccharide fatty acids and Telmisartan modified chitosan oligosaccharide fatty acid are weighed respectively, add 2mL water, Probe Ultrasonic Searching
30 times, the μ l of 2mg/mL fluorescein isothiocynates 200 are taken respectively, and room temperature lucifuge stirs 12 hours.Reaction solution is put in bag filter, thoroughly
Analysis 1 day, centrifugation, obtain the chitosan oligosaccharide fatty acid and Telmisartan modified chitosan oligosaccharide fatty acid of marked by fluorescein isothiocyanate.
In 24 porocyte culture plates, 1mL is added per hole and contains 1 × 105Individual MCF-7 cells, put 37 DEG C, 5%CO2Incubator is trained
Support 12 hours, after cell is completely adherent, 100 μ l Telmisartan modified chitosan oligosaccharide fatty acids are added per hole.It is small to continue incubation 12
When, nutrient solution is discarded, is cleaned three times with isotonic phosphate buffer, cell dissociation is got off with 0.25% pancreatin, centrifugation obtains
Cell precipitation is obtained, cell precipitation, the fluorescent value of flow cytomery fluorescein isothiocynate are resuspended with 0.5mL buffer solutions.Inspection
It is respectively 12.8 and 53.4 that result chitosan oligosaccharide fatty acid as shown in Figure 3, which is surveyed, with Telmisartan modified chitosan oligosaccharide fatty acid.
Claims (4)
1. a kind of Telmisartan modified chitosan oligosaccharide fatty acid nanoparticle, its representational general structure are：
Wherein：
A is the sugared monocyclic percentage that amino is not substituted in Telmisartan modified chitosan oligosaccharide fatty acid nanoparticle, and scope is
40%~90%,
B is the sugared monocyclic percentage that substitute by aliphatic acid in Telmisartan modified chitosan oligosaccharide fatty acid nanoparticle, scope for 1%~
50%,
C is the sugared monocyclic percentage substituted in Telmisartan modified chitosan oligosaccharide fatty acid nanoparticle by Telmisartan, and scope is
0.3%~2.0%.
A kind of 2. preparation method of Telmisartan modified chitosan oligosaccharide fatty acid described in claim 1, it is characterised in that by with
Lower step is realized：
(1) prepared by reaction solution：15mmol~30mmol Telmisartan is taken, is dissolved in 1.5mL DMFs, is added
150mmol n-hydroxysuccinimides and 150mmol 1- (3- dimethylamino-propyls) -3- ethyl-carbodiimide hydrochlorides, 60
DEG C activation 1 hour；
(2) chitosan oligosaccharide fatty acid 2mmol is weighed, is dissolved in 4mL water, Probe Ultrasonic Searching 30 times, adds above-mentioned reaction solution, and add
4.0mL DMFs, 60 DEG C are reacted 12 hours；Reaction solution is put in bag filter, dialyses 2 in the acid solutions of pH 1.0
My god, distilled water continues dialysis 2 days, freeze-drying, obtains purpose product Telmisartan modified chitosan oligosaccharide fatty acid.
A kind of 3. preparation method of Telmisartan modified chitosan oligosaccharide fatty acid according to claim 2, it is characterised in that institute
Chitosan oligosaccharide fatty acid is covered by CN200510050798.1 and CN200610051601.0, chitosan oligosaccharide fatty acid amino
Substitution value is 1%~50%, and wherein 1~200kDa of molecular weight of chitosan oligosaccharide, the carbon chain lengths of aliphatic acid are：C12-C22, shell are few
The deacetylation of sugar is 70%-100%.
4. a kind of Telmisartan modified chitosan oligosaccharide fatty acid according to claim 1 is in anti-tumor immunotherapy medicine is prepared
Application, it is characterised in that prepare targeting breast cancer tumor cells anti-tumor immunotherapy medicine in application.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101148483A (en) * | 2007-11-08 | 2008-03-26 | 浙江大学 | Chitosan oligosaccharide-stearic acid grafting compound modified by folic acid, preparation method and application thereof |
| CN101293933A (en) * | 2008-06-17 | 2008-10-29 | 浙江大学 | Polyglycol modified chitosan oligosaccharide fatty acid grafting article, preparing method and application thereof |
| CN101314622A (en) * | 2008-06-26 | 2008-12-03 | 浙江大学 | Salicylic-g-chitosan oligosaccharide grafts and synthesizing process |
| CN101524326A (en) * | 2009-03-30 | 2009-09-09 | 浙江大学 | Chlorin e6 chitosan-stearic acid graft micelle |
| CN101564539A (en) * | 2009-05-25 | 2009-10-28 | 浙江大学 | Chitosan oligosaccharide fatty acid grafting-adriamycin bonded drug, preparation and applications |
| CN102863557A (en) * | 2012-10-12 | 2013-01-09 | 苏州大学 | Preparation method and application of fatty acid-trimethyl chitosan polymer modified by lactobionic acid |
-
2017
- 2017-10-26 CN CN201711019493.3A patent/CN107875399B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101148483A (en) * | 2007-11-08 | 2008-03-26 | 浙江大学 | Chitosan oligosaccharide-stearic acid grafting compound modified by folic acid, preparation method and application thereof |
| CN101293933A (en) * | 2008-06-17 | 2008-10-29 | 浙江大学 | Polyglycol modified chitosan oligosaccharide fatty acid grafting article, preparing method and application thereof |
| CN101314622A (en) * | 2008-06-26 | 2008-12-03 | 浙江大学 | Salicylic-g-chitosan oligosaccharide grafts and synthesizing process |
| CN101524326A (en) * | 2009-03-30 | 2009-09-09 | 浙江大学 | Chlorin e6 chitosan-stearic acid graft micelle |
| CN101564539A (en) * | 2009-05-25 | 2009-10-28 | 浙江大学 | Chitosan oligosaccharide fatty acid grafting-adriamycin bonded drug, preparation and applications |
| CN102863557A (en) * | 2012-10-12 | 2013-01-09 | 苏州大学 | Preparation method and application of fatty acid-trimethyl chitosan polymer modified by lactobionic acid |
Non-Patent Citations (3)
| Title |
|---|
| FU-QIANG HU，ET AL: ""A novel chitosan oligosaccharide–stearic acid micelles for gene delivery: Properties and in vitro transfection studies"", 《INTERNATIONAL JOURNAL OF PHARMACEUTICS》 * |
| HONG YUAN,ET AL: ""stearic Acid-g-chitosan Polymeric Micelle for Oral Drug Delivery: In Vitro Transport and in Vivo Absorption"", 《MOLECULAR PHARMACEUTICS》 * |
| JIAN YOU，ET AL: ""Polymic Micelles with Glycolipid-like Structure and Multiple Hydrophobic Domains for Mediating Molecular Target Delivery of Paclitaxel"", 《BIOMACROMOLECULES》 * |
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| Publication number | Publication date |
|---|---|
| CN107875399B (en) | 2020-04-03 |
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