CN109395092A - A kind of carrier and its application based on host-guest interaction - Google Patents
A kind of carrier and its application based on host-guest interaction Download PDFInfo
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- CN109395092A CN109395092A CN201811264940.6A CN201811264940A CN109395092A CN 109395092 A CN109395092 A CN 109395092A CN 201811264940 A CN201811264940 A CN 201811264940A CN 109395092 A CN109395092 A CN 109395092A
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- 229910001414 potassium ion Inorganic materials 0.000 description 1
- BDAWXSQJJCIFIK-UHFFFAOYSA-N potassium methoxide Chemical compound [K+].[O-]C BDAWXSQJJCIFIK-UHFFFAOYSA-N 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/69—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit
- A61K47/6949—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes
- A61K47/6951—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the conjugate being characterised by physical or galenical forms, e.g. emulsion, particle, inclusion complex, stent or kit inclusion complexes, e.g. clathrates, cavitates or fullerenes using cyclodextrin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7028—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
- A61K31/7034—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
- A61K31/704—Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
- A61K48/005—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy characterised by an aspect of the 'active' part of the composition delivered, i.e. the nucleic acid delivered
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a kind of carrier based on host-guest interaction and its applications, carrier includes host molecule and guest molecule, host molecule is the cyclodextrin of polymer graft modification, and guest molecule can be contained in the ring of cyclodextrin, and guest molecule covalent coupling is on the surface of PCL-HPG.Carrier of the invention, cytotoxicity is low, all has good carrying capacity to hydrophobic drug and nucleic acid sequence, has good tolerance and cell compatibility to serum, while can improve transfection efficiency well.By the transformation of guest molecule, the carrier of different pH responses can be obtained.
Description
Technical field
The present invention relates to a kind of carriers based on host-guest interaction, while further including its application.
Background technique
In recent years, delivering chemotherapeutics and gene altogether becomes research hotspot both domestic and external for treatment of cancer.Drug and
The combination of gene can mutually promote, complement each other, and reduce chemotherapeutics dosage, and being finally reached reduces toxic side effect, improves treatment
The purpose of effect.The key of the technology successful implementation is a kind of not only safe but also efficient carrier material.Currently, there are many
Carrier material is used for the total delivering of drug and gene, such as inorganic nano-particle, liposome, micella.
Drug candesartan (CD) is coupled on polyethyleneimine (PEI) and modifies to single by Wang etc. [1]
Manage (SWNTs), then again with vascular endothelial growth factor targeting siRNA (siVEGF) occur electrostatic it is compound, formed drug and
The total delivery system (SWNT-PEI-CD/siVEGF) of gene, for cooperateing with and targeting therapy on tumor angiogenesis.Result of study
It has been shown that, SWNT-PEI-CD/siVEGF is total to delivery system can have with cancer target specificity, and by combination drug and gene
Effect inhibits Tumor Angiongesis, and therapeutic effect is better than being administered alone or gene.Wang and Chen [2] develops a kind of lipoprotein and spreads out
Biological nano carrier (rHDL), for loading therapeutic gene (p53) and hydrophobicity chemotherapeutics (PTX), studies have shown that by excellent
The PTX-DODAB/p53-rHDL nanoparticle of change can not only be mediated by SR-BI and realize excellent tumor-targeting, also to lotus
Tumor nude mice has significant antitumous effect.Zhou seminar [3] is successfully synthesized in a recent research by one kettle way
The amphiphilic copolymer of pH sensitivity and isotope of redox-sensitive, the copolymer can form micella and to DOX and PLK1 specificity
ShRNA carries out payload, sensibility can be triggered to discharge rapidly under tumour cell microenvironment the drug that contains and
Gene；And have the nude mice of U87 glioma as animal model to establish, the system is had studied to the therapeutic effect of solid tumor, discovery
The therapeutic efficiency for delivering therapy system altogether is higher than list to DOX or single to shRNA treatment mode.In order to by dewatering medicament DOX and
Functional gene pMMP-9 is effectively delivered to tumour cell, and the designs such as Ma [4] have been synthesized one kind and modified with porphyrin with core, arginine
Poly (L-lysine) be arm star-shaped cationic polymer (PP-PLLD-Arg).Studies have shown that carrying drug or gene phase with single
Than PP-PLLD-Arg/DOC/pMMP-9 compound not only can induce more significant Apoptosis, but also significantly reduce HNE-1
The invasive ability of cell.Liu [5] successfully synthesizes the knot being made of β-CD and dendroid poly (L-lysine) by click-reaction
The clearly star-like cationic copolymer (CD-PLLD) of structure.
Researchers have done many effort in terms of design prepares highefficiency non-viral carrier, from cationic-liposome to
Then linear cationic polymer arrives hyperbranched cationic polymer, but how to balance between transfection efficiency and cytotoxicity
Contradiction is still a major challenge.By taking branched polyethylene imine (PEI) as an example, molecular weight is many weeks by 25000 PEI (PEI25k)
That knows has stable and high gene transfection efficiency genophore, is typically used as evaluating other non-viral gene vector transfection efficiencies
Gold standard.However, instability limit its application clinically in higher cytotoxicity and blood.Low molecule
Amount PEI600 (PEI that molecular weight is 600) shows low cytotoxicity and good stability, but efficiency gene transfection is very
It is low.
Li Xiaohui hyperbranched poly caprolactone/poly epihydric alcohol ether copolymer synthesis and in terms of gene delivery vector
A kind of novel hyperbranched poly caprolactone/polyglycidyl ether copolymerization is disclosed using [D] China Concord Medical Science University, 2009.
Object (poly (epsilon-caprolactone)-b-hyperbranchedpolyglycidol, PCL-b-HPG).PCL-b-
HPG has amphipathic and has great amount of hydroxy group end group, can improve the hydrophily and modifiability of polycaprolactone.By PCL-b-
HPG is prepared into nano-micelle, has investigated its cytotoxicity and gene transfection, and end is coupled rgd peptide, to new to develop
Type targeted drug delivery system lays the foundation.It is observed by TEM, SEM and particle size analyzer, gained nano-micelle is spherical shape, shape
Shape is uniform, and partial size is (170 ± 10.0) nm.Using mtt assay, the cytotoxicity of PCL-b-HPG nano-micelle is had detected, and is prepared
Load pEGFP-C1 gene nano micella, the average grain diameter of prepared nano-micelle are about 225nm, and pEGFP-C1 plasmid contains
Amount accounts for about 2%, and gene embedding efficiency is 85% or more.Gel blocking electrophoresis experiment, display load base in the form of nano-micelle
There is apparent traction in swimming lane, shows that PCL-b-HPG nano-micelle can effectively wrap up plasmid DNA molecule, in vitro in cause
EA.hy926 cell transfection assays confirm PCL-b-HPG carry gene nano micella can transfected plasmids DNA, transfection efficiency about exists
15%.Its transfection efficiency or relatively low.
Therefore, how to design the non-viral gene vector with high gene transfection efficiency and low cytotoxicity is drug and base
Because of the critical issue in total delivery strategies.
Bibliography:
[1]Ding X F,Su Y J,Wang C,Zhang F R,Chen K R,Wang Y,Li M,Wang
W.Synergistic Suppression of Tumor Angiogenesis by the Co-delivering of
Vascular Endothelial Growth Factor Targeted siRNA and Candesartan Mediated by
Functionalized Carbon Nanovectors[J].AcsAppl Mater Inter,2017,9(28):23353-
23369.
[2]Wang W,Chen K,Su Y,Zhang J,Li M,Zhou J.Lysosome-Independent
Intracellular Drug/Gene Codelivery by Lipoprotein-Derived Nanovector for
Synergistic Apoptosis-Inducing Cancer-Targeted Therapy[J].Biomacromolecules,
2018,19(2):438-448.
[3]Wang P,Yu N,Wang Y,Sun H,Yang Z,Zhou S.Co-delivery of PLK1-
specific shRNA and doxorubicin via core-crosslinked pH-sensitive and redox
ultra-sensitive micelles for glioma therapy[J].J Mater Chem B,2018,6(1):112-
124.
[4]Ma D,Lin Q M,Zhang L M,Liang Y Y,Xue W.Astar-shaped porphyrin-
arginine functionalized poly(L-lysine)copolymer for photo-enhanced drug and
gene co-delivery[J].Biomaterials,2014,35(14):4357-4367.
[5]Liu T,Xue W,Ke B,Xie M Q,Ma D.Star-shaped cyclodextrin-poly(L-
lysine)derivative co-delivering docetaxel and MMP-9siRNA plasmid in cancer
therapy[J].Biomaterials,2014,35(12):3865-3872.
[6]Appel E A,del Barrio J,Loh X J,Scherman O A.Supramolecular
polymeric hydrogels[J].ChemSoc Rev,2012,41(18):6195-6214.
[7]Guo Y J,Guo S J,Ren J T,Zhai Y M,Dong S J,Wang E K.Cyclodextrin
Functionalized Graphene Nanosheets with High Supramolecular Recognition
Capability:Synthesis and Host-Guest Inclusion for Enhanced Electrochemical
Performance[J].Acs Nano,2010,4(9):5512-5512.
[8]Prochowicz D,Kornowicz A,Lewinski J.Interactions of Native
Cyclodextrins with Metal Ions and Inorganic Nanoparticles:Fertile Landscape
for Chemistry and Materials Science[J].Chem Rev,2017,117(22):13461-13501.
[9]Zhang Z,Lv Q,Gao X Y,Chen L,Cao Y,Yu S J,He C L,Chen X S.pH-
Responsive Poly(ethylene glycol)/Poly(L-lactide)Supramolecular Micelles Based
on Host-Guest Interaction[J].AcsAppl Mater Inter,2015,7(16):8404-8411.
[10]Xue M,Zhong X,Shaposhnik Z,Qu Y Q,Tamanoi F,Duan X F,Zink J I.pH-
Operated Mechanized Porous Silicon Nanoparticles[J].J Am ChemSoc,2011,133
(23):8798-8801.
Summary of the invention
It is an object of the invention to overcome the deficiencies of the prior art and provide a kind of carriers based on host-guest interaction.
It is another object of the present invention to provide the applications of above-mentioned carrier.
The technical solution used in the present invention is:
A kind of carrier based on host-guest interaction, including host molecule and guest molecule, host molecule are polymer
The cyclodextrin of graft modification, guest molecule can be contained in the ring of cyclodextrin, table of the guest molecule covalent coupling in PCL-HPG
Face.
As the further improvement of above-mentioned carrier, PCL has 4 branch chain arms, is the core of PCL-HPG.
As the further improvement of above-mentioned carrier, the number-average molecular weight of HPG is 45000~60000.
As the further improvement of above-mentioned carrier, the number-average molecular weight of PCL is 14000~16000.
As the further improvement of above-mentioned carrier, the polymer of cyclodextrin grafting fiber is PEI.
As the further improvement of above-mentioned carrier, the number-average molecular weight of PEI is 500~700.
As the further improvement of above-mentioned carrier, cyclodextrin is beta-cyclodextrin, and guest molecule is selected from benzimidazole, Buddha's warrior attendant
At least one of alkane, azobenzene.
Carrier is preparing the application in dewatering medicament and/or nucleic acid sequence carrier, wherein carrier is as described above.
As the further improvement of above-mentioned application, dewatering medicament is selected from DOX, Docetaxel DOC, taxol PTX；Nucleic acid
Plasmid, linear DNA and its compound, RNA and its compound that sequence is of length no more than 5000bp.
A kind of preparation method being loaded with dewatering medicament and/or nucleic acid sequence carrier, the host molecule and guest molecule of carrier
As described above, comprising:
1) host molecule of carrier and guest molecule are mixed, reaction obtains carrier；
2) carrier is mixed with dewatering medicament and/or nucleic acid sequence, removes unsupported dewatering medicament and/or nucleic acid sequence
Column, obtain the carrier for being loaded with dewatering medicament and/or nucleic acid sequence.
The beneficial effects of the present invention are:
Carrier of the invention, cytotoxicity is low, all has good carrying energy to hydrophobic drug and nucleic acid sequence
Power has good tolerance and cell compatibility to serum, while can improve transfection efficiency well.Pass through guest molecule
The transformation of son can obtain the carrier of different pH responses.
Detailed description of the invention
Fig. 1 is the synthetic route of carrier；
Fig. 2 is the coherent detection data of guest molecule；
Fig. 3 is PEI600, β-CD and β-CD-PEI6001H NMR spectra；
Fig. 4 be star-type polymer PCL-HPG-PEI600 structural schematic diagram (A) and1H NMR spectra (B)；
Fig. 5 is the spectrogram of various concentration PCL-HPG-BM and β-CD-PEI600 effect；
Fig. 6 is the isothermal titration curve of β-CD-PEI600 and PCL-HPG-BM interaction；
Fig. 7 is the experimental result of PEI25k/DNA and PCL-HPG-PEI600/DNA compound under different transfection conditions；
Fig. 8 is de-assembly behavior outcome of PCL-HPG-PEI600 under the conditions of weak acid pH；
Fig. 9 is the partial size of PCL-HPG-PEI600 under condition of different pH.
Specific embodiment
A kind of carrier based on host-guest interaction, including host molecule and guest molecule, host molecule are polymer
The cyclodextrin of graft modification, guest molecule can be contained in the ring of cyclodextrin, table of the guest molecule covalent coupling in PCL-HPG
Face.
As the further improvement of above-mentioned carrier, PCL has 4 branch chain arms, is the core of PCL-HPG.
As the further improvement of above-mentioned carrier, the number-average molecular weight of HPG is 45000~60000.
As the further improvement of above-mentioned carrier, the number-average molecular weight of PCL is 14000~16000.
As the further improvement of above-mentioned carrier, the polymer of cyclodextrin grafting fiber is PEI.
To avoid the cytotoxicity of PEI excessive, as the further improvement of above-mentioned carrier, the number-average molecular weight of PEI is 500
~700.
As the further improvement of above-mentioned carrier, cyclodextrin is beta-cyclodextrin, and guest molecule is selected from benzimidazole, Buddha's warrior attendant
At least one of alkane, azobenzene.This Subjective and Objective molecule can be mutually chimeric well, forms more stable Subjective and Objective body
System.Using physicochemical characteristic of the different guest molecules at different pH, the guest-host system of available difference pH response.
Carrier is preparing the application in dewatering medicament and/or nucleic acid sequence carrier, wherein carrier is as described above.
As the further improvement of above-mentioned application, dewatering medicament includes but is not limited to DOX, Docetaxel DOC, taxol
PTX；Plasmid, linear DNA and its compound, RNA and its compound that nucleic acid sequence is of length no more than 5000bp.
A kind of preparation method being loaded with dewatering medicament and/or nucleic acid sequence carrier, the host molecule and guest molecule of carrier
As described above, comprising:
1) host molecule of carrier and guest molecule are mixed, reaction obtains carrier；
2) carrier is mixed with dewatering medicament and/or nucleic acid sequence, removes unsupported dewatering medicament and/or nucleic acid sequence
Column, obtain the carrier for being loaded with dewatering medicament and/or nucleic acid sequence.
Explanation of nouns:
HPG: hyperbranched polyglycidyl ether Hyperbranched polyglycerol
BM: benzimidazole Benzimidazole
β-CD: beta-cyclodextrin β-Cyclodextrin
PCL: polycaprolactone Polycaprolactone
PEI: polyethyleneimine Polyethyleneimine
DOX: adriamycin Doxorubicin
Below with reference to embodiment, technical solution of the present invention is further illustrated.
The synthesis of carrier
The synthetic route of carrier is as shown in Figure 1.Scheme the synthetic route of (A) PCL-HPG-BM；(B) conjunction of β-CD-PEI600
At route.(C) structural schematic diagram of star-type polymer PCL-HPG-PEI600.
The synthesis of PCL-HPG
It is double using anhydrous and oxygen-free in 50mL Schlenk flask that 3.0g (0.20mmol) dry PCL is weighed first
Pipe device vacuumizes logical nitrogen for several times to guarantee no oxygen in reaction system repeatedly.Then it is molten that 0.8mL 25wt% potassium methoxide is added
Liquid is gradually heated to 95 DEG C while stirring.After 10min, vacuumizes remove extra methanol again.Then, under the protection of nitrogen,
12g (162mmol) epoxy prapanol is added dropwise into reaction system with micro-injection pump.After dripping, the reaction was continued 5h.Reaction
After, with proper amount of methanol lysate, and the cation exchange resin after acidification is added to remove potassium ion.Revolving is except removal
After solvent obtains crude product, yellow transparent thick liquid is lyophilized to obtain after the bag filter dialysis for being 3000 with molecular cut off.
The synthesis of PCL-HPG-BM
0.5g PCL-HPG and 0.05g (0.3mmol) BM is dissolved in dry n,N-Dimethylformamide (DMF),
0.02g (0.1mmol) DCC and 0.005g (0.3mmol) DMAP is then added into reaction system and to stir.React at room temperature 48h
Afterwards, yellow transparent thick liquid is lyophilized to obtain after the bag filter dialysis for being 1000 with molecular cut off by reaction solution.
Fig. 2 is the coherent detection data of guest molecule, wherein figure (A) PCL-HPG1H NMR spectra；(B)PCL-HPG-
BM, PCL-HPG, 4-arms-PCL and BM's1H NMR spectra.It can be seen that 1.3,1.6,2.3 and of chemical shift from figure A
3.99ppm is respectively the methene proton absorption peak of different location on 4-arms-PCL.And the vicinity chemical shift 3.4ppm
Proton uptake peak belongs to the methylene and methine of HPG, and the absorption peak at 4.6ppm is then attributed to the hydroxyl of HPG, this is illustrated
The successful synthesis of PCL-HPG.Raw material BM and 4-arms-PCL, the nuclear-magnetism of product PCL-HPG and PCL-HPG-BM is shown in figure
Hydrogen spectrogram can not only find returning for each proton peak in 4-arms-PCL and HPG from the nucleus magnetic hydrogen spectrum figure of PCL-HPG-BM
Belong to, the diagnostic protons absorption peak of BM can also be seen at chemical shift 7.5-8.5ppm.This illustrates guest molecule PCL-HPG-
The successful synthesis of BM.
The synthesis of β-CD-PEI600
The synthesis of β-CD-PEI600 is divided into two steps, and the first step is mono- 6- tolysulfonyl-beta-cyclodextrin (β-CD-OTs)
Synthesis, weigh 31.2g (26.4mmol) β-CD in 500mL single necked round bottom flask, be added 250mL deionized water, stirring extremely
It is evenly dispersed；Then to 10mL NaOH aqueous solution (8.2M) is slowly added dropwise in suspension.It drips after reaction solution clarification,
Reaction system is cooled to 0 DEG C；Then 20mL paratoluensulfonyl chloride acetonitrile solution (1.3M), completion of dropwise addition are slowly added dropwise thereto
Afterwards, the reaction was continued at room temperature 2h.Then, pH to 6 or so is adjusted with dilute hydrochloric acid, is collected by centrifugation and precipitates to obtain crude product, uses pure water
After recrystallizing twice, it is dried in vacuo to obtain white powder.Second step is the synthesis of β-CD-PEI600, first by 1.1g (1.9mmol)
PEI600 is dissolved in 10mL dry dimethyl sulfoxide (DMSO), and 1.9g (1.5mmol) β-CD-OTs is then added into system.
After system reaction temperature is risen to 70 DEG C, reacted 3 days under the protection of nitrogen.Finally, being with molecular cut off by reaction solution
Light yellow clear liquid is lyophilized to obtain after 2000 bag filter dialysis.
Fig. 3 is PEI600, β-CD and β-CD-PEI6001H NMR spectra.As shown, chemical shift 2.5-3.0ppm
The proton uptake peak at place belongs to PEI, and chemical shift 5.0ppm and 3.2-4.0ppm be then respectively belonging to β-CD No. 1 hydrogen and
2-6 hydrogen.1H NMR's the result shows that β-CD-PEI600 is successfully synthesized, while passing through the proton peak face of integral β-CD and PEI
Product calculates to obtain 1.5 or so β-CD of each PEI grafting.
The synthesis of supermolecule copolymer p CL-HPG-PEI600
First 0.1g β-CD-PEI600 and 0.02g PCL-HPG-BM are dissolved in 5mL DMSO, are then added dropwise to dropwise
In 20mL PBS (pH 7.4).After mixed liquor is stirred at room temperature overnight, after being dialysed with the bag filter that molecular cut off is 3000
Supermolecule copolymer p CL-HPG-PEI600 is lyophilized to obtain.
Fig. 4 be star-type polymer PCL-HPG-PEI600 structural schematic diagram (A) and1H NMR spectra (B).As shown,
The diagnostic protons peak that chemical shift is 4-arms-PCL at the peak 1.3,1.6 and 2.3ppm；And chemical shift is in 3.4-4.0ppm model
Peak in enclosing is attributed to the characteristic peak of HPG；Peak of the chemical shift within the scope of 2.5-3.0ppm belongs to PEI600；Chemical shift
The characteristic peak that peak at 5.0ppm is typical β-CD；It is difficult at 7.5ppm since BM unit is assembled by β-CD substantially
To find the characteristic peak of BM.The nuclear-magnetism is as a result, it was confirmed that assembly polymer PC L-HPG-PEI600 is successfully prepared.
The detection of product:
The host-guest interaction of β-CD and BM are studied by fluorescence spectrum, further verify assembly PCL-HPG-
The successful preparation of PEI600.As shown in figure 5, (A) PCL-HPG-BM is in the β-CD-PEI600 solution of various concentration in Fig. 5
Fluorescence emission spectrum, the concentration of PCL-HPG-BM are 0.5mg/mL；(B) under the β-CD-PEI600 solution effects of various concentration
Fluorescence intensity of the PCL-HPG-BM at 483nm.The characteristic emission wavelength of BM molecule in PCL-HPG-BM appears in 483nm
Place, with gradually increasing for β-CD-PEI600, system fluorescence intensity is gradually decreased, and illustrates that more and more BM molecules enter
In the hydrophobic cavity of β-CD；And when the mass ratio of β-CD-PEI600 and PCL-HPG-BM reaches 10:1, fluorescence intensity is almost
It no longer reduces, fluorescence peak disappears substantially, this is because β-CD molecule is completely combined with BM molecule.Pass through the result one of fluorescence spectrum
Aspect demonstrates the successful assembling of β-CD-PEI600 and PCL-HPG-BM, on the other hand also determines host molecule β-CD-
The assembling optimum quality ratio of PEI600 and guest molecule PCL-HPG-BM is 10:1.
In addition, also determining the actual mass of β-CD-PEI600 and PCL-HPG-BM assembling by Isothermal titration calorimetry
Than titration curve is as shown in Figure 6.It is calculated by the β-CD-PEI600 amount of consumption and host molecule β-CD-PEI600 and visitor has been determined
The assembling quality ratio of body molecule PCL-HPG-BM is 7.9:1.
The loading of dewatering medicament and/or DNA compound
Dewatering medicament specifically contains that steps are as follows by taking DOX as an example:
The PCL-HPG-PEI600 compound of load DOX is prepared using dialysis.Weigh 5mg DOXHCl and 100mg
PCL-HPG-PEI600 is dissolved in 5mL DMSO, and the hydrochloric acid in 5 μ L triethylamines and in DOX is added.It keeps away at room temperature
After light is stirred overnight, it is added dropwise in 20mL deionized water.Acquired solution is transferred in the bag filter that molecular cut off is 500, is kept away
Light dialyse 48h with remove DMSO and it is unsupported on DOX, PCL-HPG-PEI600/DOX compound is lyophilized to obtain.PCL-HPG-
The content of DOX is measured using UV-Visible absorption spectrum (UV-VIS) in PEI600/DOX, Detection wavelength 483nm.
Polymer PC L-HPG-PEI600 is dissolved in ultrapure water by the preparation of PCL-HPG-PEI//pMMP-9 gene composite
In be made into certain density aqueous solution, through 0.45 μm sterile filter filtering after, it is molten that aqueous solutions of polymers is added to pMMP-9
It in liquid, and mixes well, is incubated for 20-30min at room temperature.
The DNA compound used in transfection experiment is also to obtain referring to above-mentioned steps: by polymer PC L-HPG-PEI600
Genophore and DNA after mixing, are incubated for 20-30min at room temperature and obtain.
Transfection experiment:
1) first by MCF-7 cell according to every hole cell number 5 × 104Density be inoculated in 24 orifice plates, to cell fusion
When rate is up to 70%, inhales and abandon culture medium, be replaced with the fresh DMEM culture medium containing 10%FBS；
2) 37 DEG C are placed in, 5%CO2After being incubated for 2h in cell incubator, the PCL-HPG-PEI600/ of different quality ratio is added
PMMP-9 compound (content of every hole pMMP-9 is 2 μ g)；
3) culture uses the expression of inverted fluorescence microscope qualitative observation green fluorescent protein afterwards for 24 hours and takes pictures；
4) it then inhales and abandons culture medium, washed with PBS, and digested with pancreatin, cell is collected by centrifugation；
5) it is used in combination finally, cell is resuspended with appropriate PBS using the expression percentage of FCM quantitative analysis green fluorescent protein
Flow Jo7.6.1 software analysis data (each sample do three groups parallel).
PEI25k/pMMP-9 (w/w=1.3) is as a control group.
The transfection experiment result of different condition is as shown in Figure 7.In Fig. 7, under the different transfection conditions of figure (A) PEI25k/DNA and
The transfection efficiency in vitro (1: serum-free of PCL-HPG-PEI600/DNA compound；2:10%FBS is incubated for 4h；3:10%FBS is incubated
It educates for 24 hours)；(B) there are the transfection efficiency in vitro and cell of different quality ratio PCL-HPG-PEI600/DNA compound under serum condition
Toxicity (10%FBS is incubated for for 24 hours)；(C) have under serum condition after the transfection of different quality ratio PCL-HPG-PEI600/DNA compound
Fluorescence photo (10%FBS, be incubated for for 24 hours).As can be seen that the transfection of PEI25k control group is imitated in the presence of serum from figure A
Rate influence is very big, and after being incubated for 4h altogether with cell in culture medium containing 10%FBS, cell transfecting efficiency is under serum-free condition
31.5% significantly reduce to 8.0%, decline about 74%.And the cell transfecting efficiency of PCL-HPG-PEI600/pMMP-9 also by
Still there is 21% cell to be successfully transfected to the influence of serum, but under identical transfection conditions, this result is much higher than PEI25k
Control group illustrates that PCL-HPG-PEI600 is better than PEI25k as the serum tolerance of genophore.
In addition, PCL-HPG-PEI600/pMMP-9 (120:1) compound is incubated in culture medium containing 10%FBS with cell altogether
After educating for 24 hours, compared to transfection efficiency under serum-free condition, transfection efficiency is not only up to 50.5% without decline instead, and
The transfection efficiency of PEI25k control group is only 15.5%.On the basis of this experimental result, not homogeneity is also further had studied
Measure the transfection feelings after being incubated for for 24 hours altogether in culture medium containing 10%FBS with cell than lower PCL-HPG-PEI600/pMMP-9 compound
Condition, and the cytotoxicity in transfection process is assessed.As shown in panelb, when mass ratio is only 60:1, just there is 32.5%
Cell be successfully transfected, and with the increase of mass ratio, transfection efficiency increases therewith.To avoid PCL-HPG-PEI600/
PMMP-9 compound itself inhibits interference of the cell Proliferation to result, and cytotoxicity experiment chooses pEGFP conduct in transfection process
Model plasmid schemes B the results show that each mass ratio PCL-HPG-PEI600/pEGFP compound does not have obviously under experimental conditions
Cytotoxicity, the cell survival rate for reviewing PEI25k/pEGFP control group is only 37%.The corresponding fluorescence picture of each group such as schemes C
It is shown.The above results show that PCL-HPG-PEI600/pMMP-9 compound has preferable serum tolerance and cytocompatibility
Property.
The experimental data of pH response
For the pH responsiveness for verifying assembly PCL-HPG-PEI600, PCL-HPG- is further studied by fluorescence spectrum
De-assembly behavior of PEI600 under the conditions of weak acid pH.As a result as shown in figure 8, as pH >=6.5, assembly PCL-HPG-
The fluorescence intensity of PEI600 has almost no change, but when pH is 6, fluorescence intensity is obviously reduced, this may be because weak
Under the conditions of acid, the increased hydrophilicity of BM, Host-guest Recognition declines, so that BM molecule is detached from from the hydrophobic internal cavities of β-CD,
Lead to the decline of fluorescence intensity.
In addition partial size of the PCL-HPG-PEI600 assembly under condition of different pH is surveyed by nanometer laser particle size instrument
It is as shown in Figure 9 to measure result.It can be seen from the figure that PCL-HPG-PEI600 is under the conditions of pH value range 7.4-6.8, change of size
Less, about 100-110nm or so is kept；And in system pH value continue to reduce, when pH value is 6.5, PCL-HPG-
The partial size of PEI600 suddenly increases to 160nm or more, and continues that decline change of size little with pH value.Fluorescence spectrum and DLS
The result shows that PCL-HPG-PEI600 at lower ph, has a preferable pH responsiveness, and pH value response range 6.0-6.5 it
Between, meet the microenvironment of tumour cell, this is expected to realize the degradation of assembly polymer to improve drug in tumour cell
With the delivery efficiency of gene.
Claims (10)
1. a kind of carrier based on host-guest interaction, including host molecule and guest molecule, host molecule connect for polymer
The modified cyclodextrin of branch, guest molecule can be contained in the ring of cyclodextrin, it is characterised in that: guest molecule covalent coupling exists
The surface of PCL-HPG.
2. carrier according to claim 1, it is characterised in that: PCL has 4 branch chain arms, is the core of PCL-HPG.
3. carrier according to claim 1, it is characterised in that: the number-average molecular weight of HPG is 45000~60000.
4. carrier according to claim 1, it is characterised in that: the number-average molecular weight of PCL is 14000~16000.
5. carrier according to any one of claims 1 to 4, it is characterised in that: the polymer of cyclodextrin grafting fiber is PEI.
6. carrier according to claim 5, it is characterised in that: the number-average molecular weight of PEI is 500~700.
7. carrier according to any one of claims 1 to 4, it is characterised in that: cyclodextrin is beta-cyclodextrin, guest molecule choosing
From at least one of benzimidazole, adamantane, azobenzene.
8. carrier is preparing the application in dewatering medicament and/or nucleic acid sequence carrier, it is characterised in that: carrier such as claim 1
Described in~7 any one.
9. application according to claim 8, it is characterised in that: dewatering medicament is selected from DOX, Docetaxel DOC, taxol
PTX；Plasmid, linear DNA and its compound, RNA and its compound that nucleic acid sequence is of length no more than 5000bp.
10. a kind of preparation method for being loaded with dewatering medicament and/or nucleic acid sequence carrier, the host molecule and guest molecule of carrier are such as
Described in any one of claim 1~7, comprising:
1) host molecule of carrier and guest molecule are mixed, reaction obtains carrier；
2) carrier is mixed with dewatering medicament and/or nucleic acid sequence, removes unsupported dewatering medicament and/or nucleic acid sequence, obtains
To the carrier for being loaded with dewatering medicament and/or nucleic acid sequence.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110438162A (en) * | 2019-08-22 | 2019-11-12 | 广州晶欣生物科技有限公司 | A kind of modified form non-liposomal transfection reagent box and its application method |
| CN112011040A (en) * | 2020-07-20 | 2020-12-01 | 广州医科大学 | Multiple nano-delivery system and preparation method thereof |
| CN115970066A (en) * | 2022-12-29 | 2023-04-18 | 成都爱睿康乐医疗器械有限公司 | Drug-loaded nanogel biological lubricant based on host-guest interaction and preparation method and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050276841A1 (en) * | 2004-06-07 | 2005-12-15 | California Institute Of Technology | Biodegradable drug-polymer delivery system |
| CN102399369A (en) * | 2010-09-14 | 2012-04-04 | 同济大学 | Method for preparing supramolecular polymer micelle drug carriers sensitive to amino acid |
| CN105802106A (en) * | 2016-04-22 | 2016-07-27 | 同济大学 | Preparation method of supermolecular nanometer aggregate with triple responses of temperature, UV (ultraviolet) and reducing agent |
| CN105920614A (en) * | 2016-06-22 | 2016-09-07 | 佛山市高明绿化纳新材料有限公司 | Supramolecular hydrogel medicine and gene dual-carrier material and preparation method thereof |
| CN108659649A (en) * | 2018-05-03 | 2018-10-16 | 南通大学 | A kind of degradable C of selfreparing3N4The preparation method of/host and guest body thin film |
-
2018
- 2018-10-29 CN CN201811264940.6A patent/CN109395092A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20050276841A1 (en) * | 2004-06-07 | 2005-12-15 | California Institute Of Technology | Biodegradable drug-polymer delivery system |
| CN102399369A (en) * | 2010-09-14 | 2012-04-04 | 同济大学 | Method for preparing supramolecular polymer micelle drug carriers sensitive to amino acid |
| CN105802106A (en) * | 2016-04-22 | 2016-07-27 | 同济大学 | Preparation method of supermolecular nanometer aggregate with triple responses of temperature, UV (ultraviolet) and reducing agent |
| CN105920614A (en) * | 2016-06-22 | 2016-09-07 | 佛山市高明绿化纳新材料有限公司 | Supramolecular hydrogel medicine and gene dual-carrier material and preparation method thereof |
| CN108659649A (en) * | 2018-05-03 | 2018-10-16 | 南通大学 | A kind of degradable C of selfreparing3N4The preparation method of/host and guest body thin film |
Non-Patent Citations (1)
| Title |
|---|
| XIAOYAN ZHOU，ET AL: ""Construction of a High-Efficiency Drug and Gene Co-Delivery System for Cancer Therapy from a pH-Sensitive Supramolecular Inclusion between Oligoethylenimine-graft-β-cyclodextrin and Hyperbranched Polyglycerol Derivative"", 《ACS APPLIED MATERIALS & INTERFACES 》 * |
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| CN110438162A (en) * | 2019-08-22 | 2019-11-12 | 广州晶欣生物科技有限公司 | A kind of modified form non-liposomal transfection reagent box and its application method |
| CN110438162B (en) * | 2019-08-22 | 2021-06-25 | 广州晶欣生物科技有限公司 | Improved non-liposome transfection kit and application method thereof |
| CN112011040A (en) * | 2020-07-20 | 2020-12-01 | 广州医科大学 | Multiple nano-delivery system and preparation method thereof |
| CN112011040B (en) * | 2020-07-20 | 2021-08-17 | 广州医科大学 | Multiple nano-delivery system and preparation method thereof |
| CN115970066A (en) * | 2022-12-29 | 2023-04-18 | 成都爱睿康乐医疗器械有限公司 | Drug-loaded nanogel biological lubricant based on host-guest interaction and preparation method and application thereof |
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