CN109200294A - Viroid gene-drug transmits nano-carrier and its preparation method and application altogether - Google Patents
Viroid gene-drug transmits nano-carrier and its preparation method and application altogether Download PDFInfo
- Publication number
- CN109200294A CN109200294A CN201710547692.5A CN201710547692A CN109200294A CN 109200294 A CN109200294 A CN 109200294A CN 201710547692 A CN201710547692 A CN 201710547692A CN 109200294 A CN109200294 A CN 109200294A
- Authority
- CN
- China
- Prior art keywords
- nano
- gene
- drug
- dna
- viroid
- 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.)
- Pending
Links
Classifications
-
- 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
- A61K48/0066—Manipulation of the nucleic acid to modify its expression pattern, e.g. enhance its duration of expression, achieved by the presence of particular introns in the delivered nucleic acid
-
- 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
- A61K48/0008—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 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition
- A61K48/0025—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 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid
- A61K48/0041—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 'non-active' part of the composition delivered, e.g. wherein such 'non-active' part is not delivered simultaneously with the 'active' part of the composition wherein the non-active part clearly interacts with the delivered nucleic acid the non-active part being polymeric
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/141—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers
- A61K9/146—Intimate drug-carrier mixtures characterised by the carrier, e.g. ordered mixtures, adsorbates, solid solutions, eutectica, co-dried, co-solubilised, co-kneaded, co-milled, co-ground products, co-precipitates, co-evaporates, co-extrudates, co-melts; Drug nanoparticles with adsorbed surface modifiers with organic macromolecular compounds
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Epidemiology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The present invention discloses viroid gene-drug and transmits nano-carrier and its preparation method and application altogether, nano-complex is compounded to form using polycation polyethyleneimine and DNA, DOX Nanoparticle Modified is constructed into a kind of nano-carrier with viral rough surface on the surface of PEI/DNA compound by electrostatic force.In treating cancer, there are many inevitable defects, the nano-carriers can load chemotherapeutics and therapeutic gene simultaneously for monotherapy, the final combination therapy for realizing drug and gene.The present invention can be used for the preparation of anti-tumor agent, and it has many advantages, such as that bio-toxicity is low, transfection efficiency is high, treatment ability of medicine is strong and easy to operate.
Description
Technical field
The present invention relates to a kind of novel preparation methods with tumor-selective multifunctional nano preparation, belong to biological doctor
Medicine field.
Background technique
Cancer, also referred to as malignant tumour, nowadays oneself is become the significant threat of human health and service life of surviving.At present,
Clinically common anti-cancer therapies mainly include surgical operation, radiotherapy, chemotherapy and gene therapy.However it passes
The toxic and side of system is big, and tumor-targeting is poor, is also easy to generate multidrug resistance.It is swift and violent with molecular biology
The increasingly maturation of development and nanotechnology, gene therapy already become the strong means of radical cure cancer.Gene therapy face at present
The urgent problem to be solved faced is still how therapeutic gene to be made efficiently to deliver into host cell, for this reason, it may be necessary to one
Suitable carrier is as assistance.Poly-cation is always one of the non-viral gene vector of most prospect, but they
Transfection efficiency need to be improved.The compound of DNA and carrier enter cell ability be influence efficiency gene transfection it is main because
One of element, so the cellular uptake ability that people's high expectations can have a kind of strategy that can promote DNA nano-complex.In recent years
Come, as people understand virus structure and emic in-depth, is provided for the design that we carry out artificial carrier useful
Information so that by improve transmembrane transport ability come realize high level transfection become possibility.Studies have shown that many viruses,
Such as influenza virus, herpes simplex virus (HSV) and human immunodeficiency virus (HIV) etc. are covered containing one by sharp cutting edge of a knife or a sword glycoprotein
The rough surface of lid, and the nanoscale rough virus surface of this sharp cutting edge of a knife or a sword glycoprotein composition is " friendly " to cell membrane, favorably
In cell endocytic.The nano-gene carrier for constructing a viroid surface thus would be possible to realize high transfection, finally improve cancer
The therapeutic efficiency of disease.
Virus structure is understood in depth with emic recently as people, it was recognized that the rough surface of virus
Be conducive to it into cell, be designed in succession there are many viroid nano-carrier at present.For example, the simulated virus such as Yu is coarse
Small particle silicon ball is modified in big silicon ball surface to increase its degree of roughness, is prepared for the nanometer on a viroid surface by surface
Drug-loading system.Research shows that the nano silicon spheres with rough surface are easier to enter cell.Feng Jun seminar is by gold nanoparticle
It is compounded in the nano-complex surface that load has DNA by electrostatic interaction, the organic-inorganic for foring a kind of viroid surface is miscellaneous
Change nano medicament carrying system, improves the ability that polycation gene carrier enters cell, while realizing the gene therapy of cancer
With the synergistic treatment of photo-thermal therapy.The design for making a general survey of all carriers is mostly to realize that viroid nano surface is carried by hard material
The design of body, few people carry out the design of viroid nano surface carrier using soft material.
Adriamycin (DOX) be used as a kind of common clinical cancer chemotherapeutics, since its hydrophobicity is strong, toxic side effect greatly with
And the defects such as drugloading rate is low, significantly limit its application clinically.Many studies have shown that, free DOX can in recent years
To be self-assembled into nanoparticle, partial size about 10nm or so, and it is negatively charged.Such as someone is self-assembly of a kind of no-load using DOX
The nano-carrier of body.Experiment shows that the nano-carrier has 90.47% drugloading rate, and has good oncotherapy effect.
Meanwhile existing research and clinical data show that monotherapy (chemotherapy and gene therapy) inevitably has intrinsic deficiency,
The important development direction for therefore becoming oncotherapy means in recent years is used in combination in a variety for the treatment of means.By a variety of different therapy apparatuses
The treatment factor of system, is loaded in same nano-carrier and is carried to disease location, realizes that fixed point is administered simultaneously, can be with complementary mechanisms
Defect realizes treatment synergy.But the loading and pass through mechanism of drug and gene are completely different, construct high-efficiency low-toxicity and have
The total transport system of good biocompatibility becomes one of the prominent challenge of related synergistic treatment research at present.
Summary of the invention
It is an object of the invention to overcome the deficiencies in the prior art, technical problems to be solved are to provide a kind of with class disease
Malicious rough surface, gene/drug can be achieved transmit altogether, high-efficiency low-toxicity carrier-free nanometer formulation and preparation method thereof, by DOX oneself
Assemble nanometer carrier is introduced into the surface of DNA Yu polyethyleneimine (PEI) compound by electrostatic force, on the one hand changes
The smooth surface of DNA/PEI compound improves its transfection efficiency with the ability for promoting it to enter cell；Jie of another aspect DOX
Enter, so that loading while the nano-carrier realizes gene and drug, is expected to realize the gene therapy and chemotherapy of tumour
Synergistic treatment, the final efficient treatment for realizing cancer.
Viroid gene-drug transmits nano-carrier altogether, is medicament nano particle by drug self assembly, is made by electrostatic
Gene and cationic polymer are firmly compounded to form nano-complex, recycle electrostatic force that medicament nano particle is compound
On nano-complex surface, the nano-carrier with viroid rough surface of gene and drug is formed while is mounted with.
In the above-mentioned technical solutions, drug is adriamycin.
In the above-mentioned technical solutions, gene is milt DNA or p53 gene.
In the above-mentioned technical solutions, cationic polymer PEI, Mw=1800.
In the above-mentioned technical solutions, it is 140-150nm that viroid gene-drug transmits the size of nano-carrier altogether, whole
Negative potential is presented in body.
When being prepared, carry out as steps described below:
It step 1, is medicament nano particle by drug self assembly
In step 1, drug is adriamycin, doxorubicin hydrochloride is carried out desalting processing, then obtain certainly by reprecipitation method
Assembling adriamycin nano particle: using microsyringe under the conditions of being protected from light, desalination is slowly added dropwise while stirring into ultrapure water
Adriamycin stirs 5h, mixing speed 1000rpm/5min, and submitting speed is 1-5ml per minute, and temperature is room temperature 20-25
Degree Celsius.
Step 2, gene and cationic polymer are compounded to form by nano-complex by electrostatic force
In step 2, gene is milt DNA or p53 gene；Cationic polymer is PEI, Mw=1800；Using dense
The aqueous dna progress that degree is 1mg/mL PEI aqueous solution and concentration is 200ng/ μ L is compound, and the two recombination time is 10-
40min, preferably 30-40min, temperature are 20-25 degrees Celsius of room temperature.
In step 2, the N/P ratio in gene and cationic polymerization objects system is 20-30, i.e. N/P ratio refers to carrier material
Expect the ratio between the molal quantity of P contained by the molal quantity and DNA of contained N.
Step 3, medicament nano particle is compounded in nano-complex surface using electrostatic force
Adriamycin nano particle (DOX NPs) is added in the aqueous solution containing nano-complex PEI/DNA, passes through electrostatic force
Effect, so that adriamycin nano particle and PEI/DNA progress are compound, recombination time is 10-15min, and temperature is room temperature 20-25
Degree Celsius.
In step 3, the potential for first measuring DOX and PEI/DNA respectively carries out of different proportion further according to the two potential
Match and mix, obtains uncharged particle.
Viroid gene-drug of the invention transmits nano-carrier in preparation gene therapy and/or chemotherapeutic agent altogether
In application.
What the genomic medicine prepared by the present invention with viroid surface transmitted carrier selection altogether is the poly- second of low molecular weight
Alkene imines, toxicity is low, good biocompatibility.DOX NPs nanoparticle is increased by the surface that electrostatic force is compounded in PEI/DNA
Add the surface roughness of PEI/DNA nano-complex, so that carrier is easier to carry out cell endocytic, improves transfection efficiency.Meanwhile
The total transmitting of gene and drug may be implemented in total transport system prepared by the present invention, realizes gene therapy in field of cancer treatment
With the synergistic treatment of chemotherapy.Preparation method of the present invention is simple, and the low transfection efficiency of toxicity is high, and the benefit of drug and gene can be improved
With rate, the fragmentation effect to tumour cell is further increased.
Detailed description of the invention
Fig. 1 is the chemical structural formula of PEI1800 used in the present invention.
Fig. 2 is the DLS that viroid rough surface genomic medicine prepared by the present invention transmits nano-carrier PEI/DNA@DOX altogether
Image (grain size distribution).
Fig. 3 is the TEM that viroid rough surface genomic medicine prepared by the present invention transmits nano-carrier PEI/DNA@DOX altogether
Image, wherein A is DOX NPs, B PEI/DNA, C are PEI/DNA@DOX.
Fig. 4 is that viroid rough surface genomic medicine prepared by the present invention transmits nano-carrier PEI/DNA@DOX altogether
HeLa cytotoxicity image, the respectively cytotoxicity schematic diagram of PEI/DNA, DOX NPs and PEI/DNA DOX.
Fig. 5 is the laser co-focusing image after PEI/DNA is cultivated 4 hours in HeLa cell in the embodiment of the present invention,
Green is the DNA of YoYo-1 label, and blue is nucleus.
Fig. 6 is that viroid rough surface genomic medicine transmits nano-carrier PEI/DNA@DOX altogether in the embodiment of the present invention
Laser co-focusing image after cultivating 4 hours in HeLa cell, Green are the DNA of YoYo-1 label, and blue is cell
Core, red are DOX.
Specific embodiment
Technical solution of the present invention is further illustrated combined with specific embodiments below.
The preparation (DOX NPs) of adriamycin nano particle
The DOXHCl of 10mg is added in 9mL dimethyl sulfoxide (DMSO), then 1mL triethylamine is added into solution
(TEA), 12h is stirred under the conditions of being protected from light, and prepares the DOX desalting soln of 1mg/mL.
The DOX desalting soln of the 1mg/mL of 200 μ L is taken to use microsyringe under being vigorously stirred of 1000rpm/5min
It is slowly dropped in 10mL water, 12h is stirred under the conditions of being protected from light, it is filtered with 220nm filter before use.Specific preparation
It is detailed in bibliography Wang C, Cheng L, Liu Z.Drug delivery with upconversion
nanoparticles for multi-functional targeted cancer cell imaging and therapy.
[J].Biomaterials,2011,32(4):1110-20。
The preparation of PEI/DNA nano-complex
The PEI aqueous solution (1mg/mL) that molecular weight is 1800 is prepared, concentration is the aqueous dna (milt of 200ng/ μ L
DNA is bought certainly: Tianjin Kai Tong reagent Chemical Co., Ltd., specification: 5g).Then according to the N/P ratio (N/P) of PEI and DNA
For 20 nano-complex.That is: 2.6 μ gPEI are added in the DNA solution of 5.0 μ g, 92.4mL ultrapure water is added, it will be compound
Object vortex 30s is placed in 37 DEG C of environment and is incubated for 30min.Compound is diluted to 1mL with containing ultrapure water later.
The preparation of PEI/DNA@DOX nano-carrier
The DOX NPs solution of 80 μ L is added into the DNA/PEI nano-complex solution of 1mL, then vortex 30s should
Compound is placed in 37 DEG C of insulating boxs and is incubated for 15min, and it is compound to make it through electrostatic force.
The partial size of PEI/DNA@DOX nano-carrier and potentiometric analysis
Use the partial size and potential of dynamic light scattering particle size instrument (DLS) test nanoparticle.The partial size of the DOX of DOX NPs
For 10nm, PDI 0.384, potential is -34.1mV.The partial size of PEI/DNA is 140nm or so, and potential is+22.0mV.It is different dense
Partial size and potential of the DOX with PEI/DNA compound after compound are spent, so that final PEI/DNA@DOX nano-carrier is electroneutral grain
Son first measures the potential of DOX and PEI/DNA respectively, the matching and mixing of different proportion are carried out further according to the two potential, is obtained
Uncharged particle.With being continuously increased for DOX NPs liquor capacity is added, the partial size of PEI/DNA@DOX has becoming for an increase
Gesture, potential constantly reduce.The increase of partial size and the reduction of potential illustrate that DOX NPs has successfully been combined on PEI/DNA, increase
Add the volume of PEI/DNA and neutralizes its electropositive.When it is 80 μ L that volume, which is added, in DOX NPs, the potential of nanoparticle is-
5.58mV, close to electroneutral, biocompatibility is best.Its average grain diameter is 152.4nm simultaneously, also complies with the size of nanoparticle
It is required that.Enter the influence of cell and the drugloading rate of DOX to nanoparticle in view of potential and partial size, selecting DOX volume is 80
The compound of μ L further is studied as instance model.
Research of the PEI/DNA@DOX nano-carrier to HeLa cytotoxicity
Cytotoxicity of the PEI/DNA before and after compound DOX NP is detected by mtt assay, while adding adriamycin nano-particles
Son amounts to three groups of sample P EI/DNA, DOX NPs and PEI/DNA@DOX.HeLa cell is with 6.0 × 103A cell per well it is close
Degree is seeded in 96 orifice plates before complex solution is added, and it is small that 24 are cultivated in the DMEM culture medium that 100 μ L contain 10%FBS
When.Use the method described in DLS experimental section by polymer and the milt DNA of 1 μ g in N/P than the different conditions for 20
Under it is compound.And the DOX (10 μ L, 30 μ L, 50 μ L, 70 μ L, 80 μ L) of various concentration, the compound 15min in 37 DEG C of insulating boxs is added
Afterwards, after it being co-cultured 48 hours with cell, original training is substituted with the fresh DMEM culture medium containing 10%FBS of 200 μ L
Base is supported, and 20 μ L MTT (5mg/mL in PBS buffer) solution is added in every hole, continues culture 4 in 37 DEG C of incubators
Hour, culture medium is discarded, 150 μ L DMSO are added in every hole, detection should on microplate reader (Bio-Red, Model 550, USA)
Photon absorbing intensity of the solution at 570nm.The relative survival rate of cell is calculated as follows: Cell viability (%)=
(OD570(smple)/OD570(control)) × 100%, wherein OD570(sample)And OD570(Control)It respectively represents no sample processing and has
Solution light absorption value after sample treatment.
As can be seen from Figure, the cell survival rate in PEI/DNA compound is up to 98% or more, it was demonstrated that PEI/DNA is multiple
Object is closed substantially without toxicity.And due to the introducing of DOX nanoparticle, cell survival rate is decreased obviously, and with DOX nanoparticle
The increase cytotoxicity of concentration significantly increases.The toxicity for proving PEI/DNA@DOX is as caused by DOX NPs.The body of DOXNPs
Its toxicity reaches maximum when product is 80 μ L, therefore further determines that DOX NPs additional amount is 80 μ L.The cell toxicant of PEI/DNA@DOX
Property be greater than naked DOX NP, prove that the introducing of DOX NPs changes the surface roughness of nanoparticle, causes from another angle
The variation of cell endocytic behavior, finally enhances its killing ability to tumour cell.
Research of the PEI/DNA@DOX nano-carrier to the endocytosis behavior of HeLa cell and COS7 cell
Using the endocytosis behavior intracellular in HeLa cell and COS7 of laser co-focusing (CLSM) research nanoparticle.This reality
The best transfection for testing middle selection PEI and DNA is studied than 20.First by cell with 1 × 105Concentration be seeded in three individually
In small culture dish, the culture of DMEM culture medium 24 hours containing 10%FBS under the conditions of 37 DEG C.The DNA of 1 μ g is multiple with polymer
Before conjunction, the concentration with 2.5 μ L is 1 × 10-5YoYo-1 acts on 15min at 37 DEG C to carry out fluorescent marker, is then pressed
It is according to the method in PEI/DNA compound preparation experiment and DLS test experiments that it is compound with PEI and DOX NPs progress, and
It is incubated for 30min under the conditions of 37 DEG C.Then with the DMEM culture medium containing 10%FBS by DOX NP, PEI/DNA, PEI/
DNA DOX compound is diluted to 1mL and is added in the single culture dish for being inoculated with cell immediately.After culture 4 hours, training is discarded
Support base and with 500mL PBS gently rinse several times, then laser confocal microscope (Nikon C1-si TE2000,
Japan the distribution situation of feux rouges, green light and blue light is observed under), the excitation wavelength of green light is 488nm, and launch wavelength is 515-
530nm；The excitation wavelength of feux rouges is 561nm；The excitation wavelength of blue light is 405nm.Cell image passes through 3.70 software of EZ-C1
Record.In order to avoid fluorescent quenching occurs, experiment whole process is protected from light.
As it can be seen in figures 5 and 6, green represents the milt DNA of YoYo-1 label, red represents the fluorescence of DOX, and blue represents
The nucleus of 33342 labels.Image clearly is shown in the cytoplasm of green fluorescence dispersion HeLa cell, it is seen that Fig. 6 is green
Fluorescence also has certain enhancing compared with the green fluorescence in Fig. 5, illustrates that the soft material DOX NPs of surface modification is introduced into PEI/
The surface of DNA nano-complex increases its surface roughness, plays certain rush to the endocytosis behavior of PEI/DNA@DOX
Into effect.Fluorescence intensity of the red fluorescence intensity also above pure DOX NPs of composite nanoparticle simultaneously, with Fig. 4 result kissing
It closes, also further demonstrates that viroid nano gene/drug is total to the design of carrier system and can effectively improve ability into cell, most
Its anti-cancer ability is improved eventually.Enhance while Fig. 6 red fluorescence is with green fluorescence, also further illustrating the nano-carrier can be same
When by DNA and DOX delivery enter cell, also further demonstrate that DOX NPs is modified well in PEI/DNA from another side
Composite surface, the nano-carrier have while realizing the potentiality of gene therapy and chemotherapy.
Content records the adjustment for carrying out technological parameter according to the present invention, and the preparation of carrier of the present invention can be achieved and show
Almost the same performance is such as chosen for use as the DNA:p53 gene for the treatment of, buys certainly: Beijing Sai Ye Biotechnology Co., Ltd, rule
Lattice: 20 μ g prepare the carrier with treatment function.Illustrative description has been done to the present invention above, it should explanation, not
In the case where being detached from core of the invention, any simple deformation, modification or other skilled in the art can not be spent
The equivalent replacement of creative work each falls within protection scope of the present invention.
Claims (8)
1. viroid gene-drug transmits nano-carrier altogether, which is characterized in that by drug self assembly be medicament nano particle, lead to
It crosses electrostatic force and gene and cationic polymer is compounded to form nano-complex, recycle electrostatic force by medicament nano
Particle is compounded in nano-complex surface, and the nanometer with viroid rough surface for forming while being mounted with gene and drug carries
Body.
2. viroid gene-drug according to claim 1 transmits nano-carrier altogether, which is characterized in that drug is Ah mould
Element；Gene is milt DNA or p53 gene；Cationic polymer is PEI, Mw=1800.
3. viroid gene-drug according to claim 1 transmits nano-carrier altogether, which is characterized in that viroid base
The size that cause-drug transmits nano-carrier altogether is 140-150nm, whole that negative potential is presented.
4. the preparation method that viroid gene-drug transmits nano-carrier altogether, which is characterized in that carry out as steps described below:
It step 1, is medicament nano particle by drug self assembly；
Step 2, gene and cationic polymer are compounded to form by nano-complex by electrostatic force, gene and cation are poly-
Closing the N/P ratio in objects system is 20-30；
Step 3, medicament nano particle is compounded in nano-complex surface using electrostatic force, first measures medicament nano respectively
The potential of particle and nano-complex carries out the matching and mixing of different proportion further according to the two potential, obtains uncharged particle.
5. the preparation method that viroid gene-drug according to claim 4 transmits nano-carrier altogether, which is characterized in that
In step 1, drug is adriamycin, doxorubicin hydrochloride is carried out desalting processing, then obtain self assembly Ah mould by reprecipitation method
Plain nanoparticle: using microsyringe under the conditions of being protected from light, desalination adriamycin is slowly added dropwise while stirring into ultrapure water, stirs
5h, mixing speed 1000rpm/5min are mixed, submitting speed is 1-5ml per minute, and temperature is 20-25 degrees Celsius of room temperature.
6. the preparation method that viroid gene-drug according to claim 4 transmits nano-carrier altogether, which is characterized in that
In step 2, gene is milt DNA or p53 gene；Cationic polymer is PEI, Mw=1800；Use concentration for 1mg/
The aqueous dna progress that mL PEI aqueous solution and concentration are 200ng/ μ L is compound, and the two recombination time is 10-40min, preferably
30-40min, temperature are 20-25 degrees Celsius of room temperature.
7. the preparation method that viroid gene-drug according to claim 4 transmits nano-carrier altogether, which is characterized in that
In step 3, adriamycin nano particle is added in the aqueous solution containing nano-complex PEI/DNA, by electrostatic force, with
Carry out adriamycin nano particle with PEI/DNA compound, recombination time is 10-15min, and temperature is 20-25 degrees Celsius of room temperature.
8. viroid gene-drug as described in claim 1 transmits nano-carrier in preparation gene therapy and/or chemistry altogether
Application in therapeutic agent.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710547692.5A CN109200294A (en) | 2017-07-06 | 2017-07-06 | Viroid gene-drug transmits nano-carrier and its preparation method and application altogether |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710547692.5A CN109200294A (en) | 2017-07-06 | 2017-07-06 | Viroid gene-drug transmits nano-carrier and its preparation method and application altogether |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109200294A true CN109200294A (en) | 2019-01-15 |
Family
ID=64992491
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710547692.5A Pending CN109200294A (en) | 2017-07-06 | 2017-07-06 | Viroid gene-drug transmits nano-carrier and its preparation method and application altogether |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109200294A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111920947A (en) * | 2020-08-17 | 2020-11-13 | 福州大学 | Method for preparing cyanine dye mediated nucleic acid/anticancer drug compound |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012009448A2 (en) * | 2010-07-13 | 2012-01-19 | The Regents Of The University Of California | Cationic polymer coated mesoporous silica nanoparticles and uses thereof |
| CN102851304A (en) * | 2011-06-30 | 2013-01-02 | 复旦大学 | Electrostatic coating based non-viral nucleic acid ternary complex system and preparation method thereof |
| CN103305549A (en) * | 2013-06-19 | 2013-09-18 | 四川大学 | Genetic vector system of nanoparticle with multiple oxidation-reduction stimulus response as well as preparation method and application of genetic vector system |
| CN104524597A (en) * | 2014-11-27 | 2015-04-22 | 郑州大学 | Construction method and application of anthracycline antitumor drug nanometer compound based on DNA biomaterial |
| CN105561334A (en) * | 2014-10-11 | 2016-05-11 | 中国人民解放军第二军医大学 | Temperature targeting-based nanogel gene delivery compound, and preparation method and application thereof |
-
2017
- 2017-07-06 CN CN201710547692.5A patent/CN109200294A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2012009448A2 (en) * | 2010-07-13 | 2012-01-19 | The Regents Of The University Of California | Cationic polymer coated mesoporous silica nanoparticles and uses thereof |
| CN102851304A (en) * | 2011-06-30 | 2013-01-02 | 复旦大学 | Electrostatic coating based non-viral nucleic acid ternary complex system and preparation method thereof |
| CN103305549A (en) * | 2013-06-19 | 2013-09-18 | 四川大学 | Genetic vector system of nanoparticle with multiple oxidation-reduction stimulus response as well as preparation method and application of genetic vector system |
| CN105561334A (en) * | 2014-10-11 | 2016-05-11 | 中国人民解放军第二军医大学 | Temperature targeting-based nanogel gene delivery compound, and preparation method and application thereof |
| CN104524597A (en) * | 2014-11-27 | 2015-04-22 | 郑州大学 | Construction method and application of anthracycline antitumor drug nanometer compound based on DNA biomaterial |
Non-Patent Citations (2)
| Title |
|---|
| CAITONG YU等: "Smart doxorubicin nanoparticles with high drug payload for enhanced chemotherapy against drug resistance and cancer diagnosis", 《NANOSCALE》 * |
| HUI-ZHEN JIA等: "Virus-Surface-Mimicking Surface Clustering of AuNPs onto DNA-Entrapped Polymeric Nanoparticle for Enhanced Cellular Internalization and Nanocluster-Induced NIR Photothermal Therapy", 《ADVANCED SCIENCE》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111920947A (en) * | 2020-08-17 | 2020-11-13 | 福州大学 | Method for preparing cyanine dye mediated nucleic acid/anticancer drug compound |
| CN111920947B (en) * | 2020-08-17 | 2022-09-13 | 福州大学 | Method for preparing cyanine dye mediated nucleic acid/anticancer drug compound |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wang et al. | Facile synthesis of uniform virus-like mesoporous silica nanoparticles for enhanced cellular internalization | |
| Zakeri et al. | Polyethylenimine-based nanocarriers in co-delivery of drug and gene: a developing horizon | |
| Wang et al. | Colloidal stability of silk fibroin nanoparticles coated with cationic polymer for effective drug delivery | |
| Arsianti et al. | Assembly of polyethylenimine-based magnetic iron oxide vectors: insights into gene delivery | |
| Zhang et al. | Fabrication of HA/PEI-functionalized carbon dots for tumor targeting, intracellular imaging and gene delivery | |
| Long et al. | Functionalization of halloysite nanotubes via grafting of dendrimer for efficient intracellular delivery of siRNA | |
| Long et al. | Polyethyleneimine grafted short halloysite nanotubes for gene delivery | |
| Daniels et al. | Sterically stabilized siRNA: gold nanocomplexes enhance c-MYC silencing in a breast cancer cell model | |
| Zhang et al. | Mitochondria-targeting nanoplatform with fluorescent carbon dots for long time imaging and magnetic field-enhanced cellular uptake | |
| Yin et al. | Functional graphene oxide as a plasmid-based Stat3 siRNA carrier inhibits mouse malignant melanoma growth in vivo | |
| Luo et al. | Photoluminescent F-doped carbon dots prepared by ring-opening reaction for gene delivery and cell imaging | |
| Qu et al. | Glycyrrhetinic acid-modified graphene oxide mediated siRNA delivery for enhanced liver-cancer targeting therapy | |
| Liang et al. | ‘Smart’gold nanoshells for combined cancer chemotherapy and hyperthermia | |
| Carvalho et al. | Silica-based gene delivery systems: From design to therapeutic applications | |
| Wu et al. | A silica–polymer composite nano system for tumor-targeted imaging and p53 gene therapy of lung cancer | |
| CN105873569A (en) | Nanoscale carriers for the delivery or co-delivery of chemotherapeutics, nucleic acids and photosensitizers | |
| Imani et al. | Polyethylene glycol and octa-arginine dual-functionalized nanographene oxide: an optimization for efficient nucleic acid delivery | |
| Hu et al. | Aggregation-induced emission (AIE) dye loaded polymer nanoparticles for gene silencing in pancreatic cancer and their in vitro and in vivo biocompatibility evaluation | |
| Haddick et al. | Particle-size-dependent delivery of antitumoral miRNA using targeted mesoporous silica nanoparticles | |
| Cen et al. | Improving magnetofection of magnetic polyethylenimine nanoparticles into MG-63 osteoblasts using a novel uniform magnetic field | |
| Huang et al. | Therapeutic nanosystems co-deliver anticancer drugs and oncogene SiRNA to achieve synergetic precise cancer chemo-gene therapy | |
| Bai et al. | Real-time fluorescence tracking of gene delivery via multifunctional nanocomposites | |
| Acter et al. | Bowl-shaped mesoporous polydopamine nanoparticles for size-dependent endocytosis into HeLa cells | |
| Wu et al. | Recent advancement of bioinspired nanomaterials and their applications: A review | |
| He et al. | Degradation and bio-safety evaluation of mPEG-PLGA-PLL copolymer-prepared nanoparticles |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190115 |
|
| RJ01 | Rejection of invention patent application after publication |