CN106434754A - Composite containing anti-PD-1 gene and polycation and its application in treating cancer - Google Patents
Composite containing anti-PD-1 gene and polycation and its application in treating cancer Download PDFInfo
- Publication number
- CN106434754A CN106434754A CN201610823083.3A CN201610823083A CN106434754A CN 106434754 A CN106434754 A CN 106434754A CN 201610823083 A CN201610823083 A CN 201610823083A CN 106434754 A CN106434754 A CN 106434754A
- Authority
- CN
- China
- Prior art keywords
- gene
- complex
- polycation
- cell
- tumor
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
-
- 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
- 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)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Wood Science & Technology (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Medicinal Chemistry (AREA)
- Public Health (AREA)
- Biochemistry (AREA)
- Organic Chemistry (AREA)
- Biomedical Technology (AREA)
- Pharmacology & Pharmacy (AREA)
- General Engineering & Computer Science (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Plant Pathology (AREA)
- Physics & Mathematics (AREA)
- Medicinal Preparation (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to the technical field of cancer treatment by T cell, and particularly relates to a composite containing anti-PD-1 gene and polycation and its application in treating tumor; the composite includes poly-cationic polymer and anti PD-1 gene in the mass ratio of 1-100: 1. The composite is transfected to the T cell externally, and the secretion anti OD-1 antibody-T cell is prepared, and PD-1 antibody can be generated, thus helping to stop the PD-1 signal, effectively helping the exhausted T cell, and promoting the T cell treatment cancer of antitumor immunity response. Compared with the prior art, the secretion anti OD-1 antibody-T cell can effectively treat tumor and realize the tumor immunological therapy; besides, the composite is free from in-vivo toxicity.
Description
Technical field
The present invention relates to the technical field of oncotherapy, particularly to a kind of compound containing anti-PD-1 gene and polycation
Thing and its application in treatment tumor and in particular to one kind by anti-PD-1 (programmed death-1) gene and poly- sun from
The nano-complex that son is formed passes through in-vitro transfection to T cell, and preparation carries the anti-PD-1 antibody-T-cell of secretion, can produce PD-1
Antibody, thus helping block PD-1 signal, can effectively help exhaustion T cell, promote the T cell treatment of anti-tumor immune response swollen
The method of tumor.
Background technology
T lymphocyte target killing expresses the tumor cell of this specific antigen.Because it has target killing tumor cell
Feature, CTL therapy become the ideal scheme of tumor individual therapy, is increasingly subject to the weight of immunologists in the world
Depending on.But because the generation of PD-1 leads to T cell to lose activity.We have proposed in vitro directly general in order to solve this difficult problem
The channel genes somatic cell of anti-PD-1, thus T cell itself produces the antibody of anti-PD-1, to improve the activity of T cell, to play and control
Treat the effect of tumor.
Gene, because of its unique target spot specificity, structure designability and metabolism safety, becomes scientific research circle and generally has an optimistic view of
New medicine.However, in 20 years of past, it is defeated in vivo for gene that research worker devises a series of carrier
Send, but only only a few enters clinical stage, so far, do not have a carrier to obtain FDA certification yet, the lacking an of effective carrier
Position, leads to conveying in nucleic acid substances body to be still within the clinical bottleneck phase.
Nucleic acid substances are transported in endochylema or the nucleus of target cell, need to overcome a series of internal conveying barrier,
Therefore, efficiently whether nucleic acid carrier, be that can it be used for the key point of clinical treatment.The carrier of nucleic acid substances conveying is generally
Two categories below：(1) viral vector：Viral vector (as slow virus carrier, adenovirus vector) as gene delivery vehicles although
Have higher in-vitro transfection activity, however, its immunogenicity and the shortcoming being easily caused mutation bring for clinical trial huge
Safety problem is so that its application is limited.(2) non-virus carrier：The advantage of non-virus carrier essentially consists in, and is ensureing expected turning
Under conditions of dye activity, the immunogenicity that viral vector brought and many inflammatory reactions can be substantially reduced, its generally with
Lower two kinds of carriers design：(a) cationic-liposome；(b) polycation gene carrier.And study at present and more focus primarily upon
Polycation gene carrier and the modification of cationic-liposome, are allowed to be applied to the targeted of genetic stew.Cation lipid
Body has higher inside and outside transfection activity, however, because the positive charge on surface affects its internal normal distribution, meanwhile, by
In selecting cation lipid, immunogenicity and inflammatory reaction also become in animal experiment one of inevitable shortcoming (Gao,
K.&Huang,L.Nonviral methods for siRNA delivery.Molecular pharmaceutics 6,651-
658(2008).).
Content of the invention
For defect of the prior art, it is an object of the invention to provide a kind of containing anti-PD-1 gene and polycation
Complex and its purposes in treatment tumor.
The purpose of the present invention is achieved through the following technical solutions：
In a first aspect, the invention provides a kind of complex containing anti-PD-1 gene and polycation, described complex bag
Including mass ratio is 1～100：1 polycationic polymer and anti-PD-1 gene.
Preferably, the mass ratio of described polycationic polymer and anti-PD-1 gene is 5～50：1.
Preferably, the mass ratio of described polycationic polymer and anti-PD-1 gene is 10～30：1.
Preferably it is characterised in that described polycationic polymer be polyethyleneimine, the derivant of polyethyleneimine,
At least one in poly- spermine or polycationic polypeptides.
Preferably, described polyethyleneimine include L-PEI, in dendritic polyethyleneimine at least one
Kind；Described poly- spermine is the polycationic polymer that spermine or spermidine are formed with bridging agent；Described polycationic polypeptides include
At least one in polylysine, poly arginine or polyglutamic acid.
Preferably, described bridging agent is glutaraldehyde, terephthalaldehyde, m-terephthal aldehyde, o-phthalaldehyde(OPA), 2,5- imidazoles two
Formaldehyde, 2,4- pyridine dicarbaldehyde, 2,5- pyridine dicarbaldehyde, 2,6- pyridine dicarbaldehyde, 2,5- pyridazine dicarbaldehyde or 2,6- pyridazine two
One of formaldehyde.
Second aspect, the invention provides a kind of preparation method containing anti-PD-1 gene and the complex of polycation, bag
Include following steps：
The aqueous solution of described polycationic polymer and the aqueous solution of anti-PD-1 gene, divide in room temperature standing 20-60
Clock can be prepared by described complex.
The third aspect, the invention provides a kind of complex containing anti-PD-1 gene and polycation treatment tumor in
Application.
Fourth aspect, the invention provides a kind of complex therapies tumor containing anti-PD-1 gene and polycation for utilization
Method, comprises the following steps：
The described complex containing anti-PD-1 gene and polycation is co-cultured with T cell, preparation carries the anti-PD-1 of secretion
Antibody-T-cell, the more anti-PD-1 antibody-T-cell of secretion will be carried be expelled in tumour patient body.
Compared with prior art, the present invention has following beneficial effect：
1st, the secretion of the present invention anti-PD-1 antibody-T-cell can effectively treatment tumor, the immunization therapy of achievable tumor, and
And, there is not toxicity in vivo.
2nd, the method that the complex transfecting T cells containing anti-PD-1 gene and polycation of present invention preparation treat tumor,
Can achieve the conveying of anti-PD-1 gene efficient low toxicity, the complex that especially poly- spermine cationoid is formed with anti-PD-1 gene by
It is the function of full-time cohesion gene in vivo in spermine, thus effectively improving transfection.
Brief description
The detailed description with reference to the following drawings, non-limiting example made by reading, the further feature of the present invention,
Objects and advantages will become more apparent upon：
Fig. 1 is the particle diameter schematic diagram of the complex of the present invention；
Fig. 2 is the zeta current potential schematic diagram of the complex of the present invention；
Fig. 3 is the transmission electron microscope schematic diagram of the complex of the present invention；
Fig. 4 is the gel electrophoresis schematic diagram of the complex of the present invention；
Fig. 5 is the cytotoxicity schematic diagram of the poly- spermine of the present invention；
Fig. 6 is the schematic diagram of mouse tumor model and tumor tissue section；Wherein, figure A-D is the tumor tissues taking off, A
Tumor for physiological saline group；B is the tumor of naked DNA group；The tumor that C organizes for poly- spermine polymer (PSA)；D is the present invention's
The tumor of nano-complex group.
Specific embodiment
With reference to specific embodiment, the present invention is described in detail.Following examples will be helpful to the technology of this area
Personnel further understand the present invention, but the invention is not limited in any way.It should be pointed out that the ordinary skill to this area
For personnel, without departing from the inventive concept of the premise, some deformation can also be made and improve.These broadly fall into the present invention
Protection domain.
Following examples provide a kind of complex containing anti-PD-1 gene and polycation, and described complex includes quality
Than for 1～100：1 polycationic polymer and anti-PD-1 gene.
The mass ratio of described polycationic polymer and anti-PD-1 gene is 5～50：1.
The mass ratio of described polycationic polymer and anti-PD-1 gene is 10～30：1.
Described polycationic polymer is that polyethyleneimine, the derivant of polyethyleneimine, poly- spermine or polycation are many
At least one in peptide.
Described polyethyleneimine includes L-PEI, at least one in dendritic polyethyleneimine；Described
Poly- spermine is the polycationic polymer that spermine or spermidine are formed with bridging agent；Described polycationic polypeptides include poly- bad ammonia
At least one in acid, poly arginine or polyglutamic acid.
Described bridging agent be glutaraldehyde, terephthalaldehyde, m-terephthal aldehyde, o-phthalaldehyde(OPA), 2,5- imidazoles dicarbaldehyde, 2,
In 4- pyridine dicarbaldehyde, 2,5- pyridine dicarbaldehyde, 2,6- pyridine dicarbaldehyde, 2,5- pyridazine dicarbaldehyde or 2,6- pyridazine dicarbaldehyde
A kind of.
The described preparation method containing anti-PD-1 gene and the complex of polycation, comprises the following steps：
The aqueous solution of described polycationic polymer and the aqueous solution of anti-PD-1 gene, divide in room temperature standing 20-60
Clock can be prepared by described complex.
The application in treatment tumor of the described complex containing anti-PD-1 gene and polycation.
The described method utilizing containing anti-PD-1 gene and the complex therapies tumor of polycation, comprises the following steps：
The described complex containing anti-PD-1 gene and polycation is co-cultured with T cell, preparation carries the anti-PD-1 of secretion
Antibody-T-cell, the more anti-PD-1 antibody-T-cell of secretion will be carried be expelled in tumour patient body.
The preparation of the formation complex of the poly- spermine of embodiment 1 polycationic polymer (PSA) and anti-PD-1 plasmid DNA
By spermine and bridging agent 2, the poly- spermine of 6- pyridine dicarbaldehyde preparation is made into aqueous solution, and concentration is 2 μ g/ μ l, with going out
Bacterium water is diluted to 20ng/ μ l, 40ng/ μ l, 100ng/ μ l, 200ng/ μ l, 1000ng/ μ l and 2000ng/ μ l solution.In addition will resist
PD-1 gene plasmid DNA is made into aqueous solution, and concentration is 20ng/ μ l.Poly- spermine aqueous solutions of polymers equal-volume is added to plasmid
Aqueous dna, vortex 5min, and stand 20min, obtain final product a series of solution of nano-complexes.
The particle diameter of formation complex and the electricity of the poly- spermine of embodiment 2 polycationic polymer (PSA) and anti-PD-1 plasmid DNA
The sign of position
Put into the survey of BIC90plus granularity potentiometric analyzer using a series of complex solutions of embodiment 1 method preparation
In experimental tank, the mass ratio 0.25,0.5,2,5,10,50 of PSA and plasmid DNA in the complex of this series of complex solution, if
Put test temperature and be 25 DEG C, medium is water, viscosity is 0.89cp, refractive index is 1.330, light scattering angle is 90., Detection wavelength
For 659nm, each sample test 3 times, each run time is 2min, records mean particle size.Result is as shown in figure 1, result
The particle diameter of display nano-complex is in below 200nm.Its dispersion index all meets the requirements.
BIC90plus granularity potentiometric analyzer will be put into using a series of complex solutions prepared by embodiment 1 method
In test trough, the mass ratio 0 of PSA and plasmid DNA in the complex of this series of complex solution:1、10:1、20:1、30:1、
40:1、50:1, setting test temperature is 25 DEG C, and medium is water, and viscosity is 0.89cp, and refractive index is 1.330, and dielectric constant is
78.54, pH value is 7.0, zeta potentiometric analysiss model is Smoluchowski equation, and each sample test 3 times is transported every time automatically
Row 10 times, records zeta current potential meansigma methodss.Result is as shown in Figure 2.Knowable to the result of Fig. 2, potential all meets the requirements.
The formation complex of embodiment 3 polycationic polymer poly- spermine polymer (PSA) and anti-PD-1 plasmid DNA saturating
Radio mirror is tested
It is 1 that preparation method according to embodiment 1 to prepare PSA/ anti-PD-1 gene plasmid DNA mass ratio:The complex of l is received
Rice complex solution, sample size is 100 μ L.Draw 10 μ l, Deca, on the copper mesh of 400 mesh, is dried naturally, finally using transmission
Ultramicroscope, to observe the form of sample, records transmission electron microscope picture.Result is as shown in figure 3, result shows nano-complex
Particle diameter is in below 200nm.
The formation complex of embodiment 4 polycationic polymer poly- spermine polymer (PSA) and anti-PD-1 plasmid DNA solidifying
Gel electrophoresis are tested
Weigh 1.0g agarose, add 100ml 1 × TAE buffer, heating for dissolving in microwave oven, treat that temperature is down to 65
DEG C, add Ethidum Eremide (EB), be configured to 1.0% agarose solution (containing 0.5 μ g/ml Ethidum Eremide), pour in glue groove, insert
Enter sample comb, it is solid that room temperature places the gelling such as 0.5-1 hour.Then, extract sample comb, in electrophoresis tank, add TAE buffer not have
Gel, waits loading.Then, poly- spermine polymer and genetic stew different quality are prepared according to the preparation method of embodiment 1
The nano-complex aqueous solution of ratio, PSA is followed successively by 0 with the mass ratio of genetic stew：1、0.2：1、0.5：1、1：1、5：1、10：1、
15：1、20：1、30：1、50：1.Marker selects DSTM5000 (100 5000bp), loading 2 μ l；6 × sample-loading buffer (bromine phenol
Orchid-glycerol indicator, containing 0.25% bromjophenol blue, 40% glycerol) 1 μ l uniformly mixed with nano-complex water 5 μ l, loading 6 μ l.?
Electrophoresis 45 minutes under 110mV voltage, finally, are placed in ultraviolet gel imaging system and observe and record electrophoretogram, result such as Fig. 4 institute
Show.From electrophoretogram as can be seen that the plasmid DNA containing anti-PD-1 gene is when mass ratio is 0.5, complex is in the swimming of electric field
Blocked completely.From picture it is observed that increase with mass ratio, the swimming of complex is all blocked.
This test shows that PSA has the ability that preferable cohesion plasmid DNA forms complex, is effectively protected plasmid DNA.
The T cell toxicity detection of the poly- spermine of embodiment 5 (PSA)
Cytotoxicity is measured using mtt assay, investigates cytotoxicity from T cell, turn 96 with the cell density in 8000/hole
Porocyte plates, are placed in overnight incubation in 37 DEG C of 5% cell culture incubator.Prepare 1,2,4,8, a series of variable concentrations of 15mg/mL
Poly- spermine (being prepared by embodiment 1) solution, every hole adds 100 μ L, and diluent media is DMEM high glucose medium (serum-free no phenol
Red), take out 96 porocyte plates from incubator, suck culture fluid, every hole is rinsed once with 100 μ L phosphate buffered solution, then
Discard phosphate buffered solution, the poly- spermine polymer solution of variable concentrations is added sequentially in cell plates, parallel assay 3
Hole.It is placed in and cultivate 4 hours in cell culture incubator.Then, suck culture fluid, every hole rinses one with 100 μ L phosphate buffered solution
Secondary, then discard phosphate buffered solution, every hole adds 100 μ L DMEM high glucose medium (serum-free is no phenol red) and 25 μ L MTT
Solution (5mg/mL), continues at culture in incubator.Afterwards, suck culture fluid within 6 hours, every hole adds 100 μ L dimethyl sulfoxide,
Place fully molten first a ceremonial jade-ladle, used in libation, adopt the multi-function microplate reader determination sample in the absorbance at 570nm and 630nm (at 630nm to be
Comparison).Result shows that the toxicity of poly- spermine polymer (PSA) is relatively low, and the toxicity of high molecular PEI (molecular weight 25000) is bright
Aobvious is higher than PSA.
The anti-PD-1 gene of embodiment 6 present invention and polycationic polymer form complex transfecting T cells treatment tumor
Effect test
BALB/c Female nude mice (5 week old, body weight 20 ± 2g), raises 1 week under SPF level environment, takes SMMC7721 to grow
The passage cell of good state, by cell number 4 × 106Individual/ml Cell sap, 0.1ml is inoculated in BALB/c mouse hind leg nearly back skin
Under.Continue to raise 2 weeks, using vernier caliper measurement and observe the upgrowth situation of tumor, to calculate tumor according to equation below
Volume：Gross tumor volume (mm3)=(length × wide × wide)/2.When gross tumor volume reaches 150-200mm3When (do tumor model move
Thing is tested), it is divided into the tumor that A is physiological saline group, B is the tumor of naked DNA group, C is swelling that poly- spermine polymer (PSA) is organized
Tumor, D is the tumor of the nano-complex group of the present invention.Wherein, physiological saline group injecting normal saline, the injection of naked DNA group is naked
DNA, poly- spermine polymer (PSA) group injection PSA, the nano-complex group injection of the present invention be：Make through the embodiment of the present invention 1
(PAS is 1 with the mass ratio of anti-PD-1 plasmid DNA to standby nano-complex:40) T cell after transfecting, the T cell after this transfection
It is to secrete anti-PD-1 antibody-T-cell containing carrying.
Put to death mice after three weeks, take tumor tissues, and measure the volume size detection result of tumor as shown in fig. 6, wherein,
A-D is the tumor tissues taking off, and A is the tumor of physiological saline group, and B is the tumor of naked DNA group, and C is poly- spermine polymer (PSA)
The tumor of group, D is the tumor of the nano-complex group of the present invention；Experimental result shows：The nano-complex group treatment of the present invention
Effect fine.
The system of the formation complex of embodiment 7 polycationic polymer polyethyleneimine (PEIS) and anti-PD-1 plasmid DNA
Standby, sign, toxicity, complex transfecting T cells are become to treat the effect test of tumor
The poly- spermine that small molecule PEI (weight average molecular weight 800-1200) is prepared with bridging agent 2,6- pyridine dicarbaldehyde is polymerized
Thing is made into aqueous solution, concentration be 2 μ g/ μ l, with aquesterilisa be diluted to 20ng/ μ l, 40ng/ μ l, 100ng/ μ l, 200ng/ μ l,
1000ng/ μ l and 2000ng/ μ l solution.In addition anti-PD-1 gene plasmid DNA is made into aqueous solution, concentration is 20ng/ μ l.Will
PEIS aqueous solutions of polymers equal-volume adds to plasmid DNA aqueous solution, vortex 5min, and stands 20min, obtains final product a series of nanometers
The solution of complex.
The particle diameter of the present embodiment gained nano-complex is substantially the same manner as Example 2 with the sign of current potential, and transmission electron microscope is surveyed
Examination and gel electrophoresis test result essentially identical with embodiment 3 and 4, and the T cell toxicity of PEI toxicity is relatively low after testing, with PSA
Toxicity suitable.
Treat effect test result and the embodiment 6 of tumor using nano-complex transfecting T cells manufactured in the present embodiment
Unanimously, show that nano-complex manufactured in the present embodiment has good therapeutic effect.
Concrete application approach of the present invention is a lot, and the above is only the preferred embodiment of the present invention.It should be pointed out that more than
Embodiment is merely to illustrate the present invention, and is not limited to protection scope of the present invention.Common skill for the art
For art personnel, under the premise without departing from the principles of the invention, some improvement can also be made, these improvement also should be regarded as this
Bright protection domain.
Claims (9)
1. a kind of complex containing anti-PD-1 gene and polycation it is characterised in that described complex include mass ratio be 1～
100：1 polycationic polymer and anti-PD-1 gene.
2. contain as claimed in claim 1 the complex of anti-PD-1 gene and polycation it is characterised in that described polycation
The mass ratio of polymer and anti-PD-1 gene is 5～50：1.
3. contain as claimed in claim 2 the complex of anti-PD-1 gene and polycation it is characterised in that described polycation
The mass ratio of polymer and anti-PD-1 gene is 10～30：1.
4. the complex containing anti-PD-1 gene and polycation as described in any one of claim 1-3 is it is characterised in that described
Polycationic polymer is at least in polyethyleneimine, the derivant of polyethyleneimine, poly- spermine or polycationic polypeptides
Kind.
5. contain as claimed in claim 4 the complex of anti-PD-1 gene and polycation it is characterised in that described polyethylene
Imines includes L-PEI, at least one in dendritic polyethyleneimine；Described poly- spermine is spermine or spermidine
The polycationic polymer being formed with bridging agent；Described polycationic polypeptides include polylysine, poly arginine or polyglutamic acid
In at least one.
6. the complex containing anti-PD-1 gene and polycation as claimed in claim 5 is it is characterised in that described bridging agent is
Glutaraldehyde, terephthalaldehyde, m-terephthal aldehyde, o-phthalaldehyde(OPA), 2,5- imidazoles dicarbaldehyde, 2,4- pyridine dicarbaldehyde, 2,5- pyrrole
One of pyridine dicarbaldehyde, 2,6- pyridine dicarbaldehyde, 2,5- pyridazine dicarbaldehyde or 2,6- pyridazine dicarbaldehyde.
7. the preparation method containing anti-PD-1 gene and the complex of polycation as described in a kind of any one as claim 1-6,
It is characterized in that, comprise the following steps：
The aqueous solution of described polycationic polymer and the aqueous solution of anti-PD-1 gene, in room temperature standing 20-60 minute be
Described complex can be obtained.
8. the complex containing anti-PD-1 gene and polycation as described in a kind of any one as claim 1-6 treatment tumor
In application.
9. a kind of complex therapies tumor containing anti-PD-1 gene and polycation utilizing as described in any one of claim 1-6
Method it is characterised in that comprising the following steps：
The described complex containing anti-PD-1 gene and polycation and T cell are co-cultured, preparation carry the anti-PD-1 antibody of secretion-
T cell, the more anti-PD-1 antibody-T-cell of secretion will be carried be expelled in tumour patient body.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610823083.3A CN106434754A (en) | 2016-09-13 | 2016-09-13 | Composite containing anti-PD-1 gene and polycation and its application in treating cancer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610823083.3A CN106434754A (en) | 2016-09-13 | 2016-09-13 | Composite containing anti-PD-1 gene and polycation and its application in treating cancer |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106434754A true CN106434754A (en) | 2017-02-22 |
Family
ID=58168911
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610823083.3A Pending CN106434754A (en) | 2016-09-13 | 2016-09-13 | Composite containing anti-PD-1 gene and polycation and its application in treating cancer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106434754A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107007570A (en) * | 2017-06-13 | 2017-08-04 | 复旦大学附属金山医院 | A kind of miR 146a nano-particles and its application in the medicine for preparing treatment diabete peripheral herve pathology |
| WO2021095599A1 (en) * | 2019-11-13 | 2021-05-20 | 国立大学法人京都大学 | Pd-1 signal inhibitor combination therapy |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103328542A (en) * | 2011-01-06 | 2013-09-25 | 金拓 | Cationic polymers formed from amino group-bearing monomers and heterocyclic linkers |
| CN104257630A (en) * | 2014-09-26 | 2015-01-07 | 上海交通大学 | Polycationic nucleic acid compound nano-particles as well as preparation method and application thereof |
| CN105085680A (en) * | 2014-05-23 | 2015-11-25 | 复旦大学 | Humanized anti-PD-1 and c-MET bispecific antibody, and preparation method and application thereof |
-
2016
- 2016-09-13 CN CN201610823083.3A patent/CN106434754A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103328542A (en) * | 2011-01-06 | 2013-09-25 | 金拓 | Cationic polymers formed from amino group-bearing monomers and heterocyclic linkers |
| CN105085680A (en) * | 2014-05-23 | 2015-11-25 | 复旦大学 | Humanized anti-PD-1 and c-MET bispecific antibody, and preparation method and application thereof |
| CN104257630A (en) * | 2014-09-26 | 2015-01-07 | 上海交通大学 | Polycationic nucleic acid compound nano-particles as well as preparation method and application thereof |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107007570A (en) * | 2017-06-13 | 2017-08-04 | 复旦大学附属金山医院 | A kind of miR 146a nano-particles and its application in the medicine for preparing treatment diabete peripheral herve pathology |
| WO2021095599A1 (en) * | 2019-11-13 | 2021-05-20 | 国立大学法人京都大学 | Pd-1 signal inhibitor combination therapy |
| CN114728068A (en) * | 2019-11-13 | 2022-07-08 | 国立大学法人京都大学 | Combination therapy with PD-1 signaling inhibitors |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Yin et al. | Stem cell-based gene therapy activated using magnetic hyperthermia to enhance the treatment of cancer | |
| Park et al. | Multi-modal transfection agent based on monodisperse magnetic nanoparticles for stem cell gene delivery and tracking | |
| Barminko et al. | Encapsulated mesenchymal stromal cells for in vivo transplantation | |
| CN111603454A (en) | Multi-targeting fusion cell membrane modified bionic nano delivery system and preparation method and application thereof | |
| Tang et al. | A new method of wound treatment: targeted therapy of skin wounds with reactive oxygen species-responsive nanoparticles containing SDF-1α | |
| CN106399380A (en) | Compound containing anti-PD (Program Death)-1 gene and poly-spermidine and application of compound to treatment of tumors | |
| Qu et al. | A Universal Platform for High‐Efficiency “Engineering” Living Cells: Integration of Cell Capture, Intracellular Delivery of Biomolecules, and Cell Harvesting Functions | |
| Tzeng et al. | Polymeric nanoparticle‐based delivery of TRAIL DNA for cancer‐specific killing | |
| CN106890343B (en) | Targeting polypeptide nano gene vector compound | |
| Wu et al. | Light-responsive charge-reversal nanovector for high-efficiency in vivo CRISPR/Cas9 gene editing with controllable location and time | |
| Wang et al. | Multistage sensitive nanoCRISPR enable efficient intracellular disruption of immune checkpoints for robust innate and adaptive immune coactivation | |
| Zhou et al. | Anti‑CD206 antibody‑conjugated Fe3O4‑based PLGA nanoparticles selectively promote tumor‑associated macrophages to polarize to the pro‑inflammatory subtype | |
| Zhang et al. | Role and mechanism of neural stem cells of the subventricular zone in glioblastoma | |
| Kim et al. | Sticky “delivering-from” strategies using viral vectors for efficient human neural stem cell infection by bioinspired catecholamines | |
| CN102397554A (en) | Tumor-targeting double-drug carrying and delivery system and preparation method thereof | |
| CN106362163A (en) | Nucleic acid delivery nano systems having double-targeting function | |
| He et al. | Sonogenetic nanosystem activated mechanosensitive ion channel to induce cell apoptosis for cancer immunotherapy | |
| CN106434754A (en) | Composite containing anti-PD-1 gene and polycation and its application in treating cancer | |
| Han et al. | Three-dimensional-cultured MSC-derived exosome with hydrogel for cerebral ischemia repair | |
| Zhao et al. | X‐ray‐guided in situ genetic engineering of macrophages for sustained cancer immunotherapy | |
| Lin et al. | A hierarchical tumor-targeting strategy for eliciting potent antitumor immunity against triple negative breast cancer | |
| Venkatesan et al. | Synthesis and characterization of chitosan tripolyphosphate nanoparticles and its encapsulation efficiency containing Russell's viper snake venom | |
| Liu et al. | Amphiphilic core–shell nanoparticles containing dense polyethyleneimine shells for efficient delivery of microrNa to Kupffer cells | |
| CN102786675B (en) | Block copolymer and synthesis method thereof, and preparation method of nano particles | |
| Zhang et al. | Programmable bispecific nano-immunoengager that captures T cells and reprograms tumor microenvironment |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170222 |
|
| RJ01 | Rejection of invention patent application after publication |