CN105420277A - Polyamid-amine hyperbranched gene carrier and preparation method and application thereof - Google Patents
Polyamid-amine hyperbranched gene carrier and preparation method and application thereof Download PDFInfo
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- 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
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Abstract
The invention discloses a polyamid-amine hyperbranched gene carrier. The gene carrier is prepared from a material A, a material B, a material C and triethylamine. The molecular formula of the material A is C2(n-1)H(5n-2)Nn, wherein n is equal to 2, 3, 4 or 5. Please see the structural formula of the material C in the specification, wherein n is equal to 0, 1, 2, 3, 4, 5 or 6, R is equal to the sum of CmH2m+1, and m is equal to 1, 2, 3, 4 or 5. Please see the structural formula of the material B in the specification, wherein n is equal to 1, 2, 3, 4, 5 or 6; or please see the other structural formula of the material B in the specification, wherein x and y are equal to 1, 2, 3, 4, 5 or 6. The invention further discloses a preparation method and application of the polyamid-amine hyperbranched gene carrier. The polyamid-amine hyperbranched gene carrier is good in branching degree and is small in toxicity and good in buffering capacity when used as gene carriers, the polyamid-amine hyperbranched gene carrier can be used for anti-cancer treatment, raw material cost is low, and the preparation method is simple.
Description
Technical field
The present invention relates to polyamide-amide genophore technical field, particularly relate to hyperbranched genophore of a kind of polyamide-amide and its preparation method and application.
Background technology
Along with developing rapidly of synthetic chemistry, many new molecular structures can design search polymkeric substance topological framework, and hyperbranched polymer has become a current study hotspot due to the structure of its uniqueness and physicochemical property; Hyperbranched polymer is widely used in bio-sensing, is separated, battery power, gene delivery, drug carrier, hydrogel, carrier of photocatalyst and biomedicine etc., because of the constructional feature of the special 3 D stereo of hyperbranched polymer, can be synthesized by two radiation and convergences, two kinds of methods.Hyperbranched polymer is because have unique physics and chemistry characteristic, and as low viscosity, high-dissolvability, high-compatibility, hyperergy, hyperbranched polymer more and more obtains the close attention of scientist.In the past few decades, large quantifier elimination all concentrates on hyperbranched polymer.
Current hyperbranched polymer is becoming the non-virus carrier of the gene delivery of a new generation, in the non-virus carrier of gene delivery, hyperbranched polymer has two salient features: one is structure-controllable, and one is that chemical property can adapt to various requirement as medicine or gene delivery.The preparation of several years hyperbranched polymers in the past has obtained sizable concern.Non-viral gene vector particularly cationic polymers also receives increasing concern.Current cationic polymers polyamide-amide degree of branching is poor, large as toxicity during genophore, and surge capability is poor, and raw materials cost is higher.
Summary of the invention
The present invention proposes hyperbranched genophore of a kind of polyamide-amide and its preparation method and application, degree of branching is good, little as toxicity during genophore, and surge capability is good, can be used as anticancer therapy, and raw materials cost is low, and preparation method is simple.
The hyperbranched genophore of a kind of polyamide-amide that the present invention proposes, its raw material comprises A material, B material, C material and triethylamine;
A material molecular formula is: C
2 (n-1)h
5n-2n
n, wherein n=2,3,4,5;
C material structure formula is:
wherein n=0,1,2,3,4,5,6, R=C
mh
2m+1, m=1,2,3,4,5;
B material structure formula is:
wherein n=1,2,3,4,5,6; Or
wherein x=y=1,2,3,4,5,6.
Material A has two effects, and one is ammonolysis, and one adds end-blocking to terminal olefin by Michael.
Preferably, A material is that one or more of the polyethylene polyamine of 250-300 form by quadrol, three second tetramines and molecular weight, and B material is N, N '-methylene-bisacrylamide, and C material is Cys methyl ester hydrochloride.
Preferably, the mol ratio of N, N '-methylene-bisacrylamide and Cys methyl ester hydrochloride is the mol ratio 1:80-120 of 2-4:1-3, N, N '-methylene-bisacrylamide and A material.
Preferably, also comprise solvent, solvent by volume part comprises water 1-6 part, DMSO0-5 part.
Preferably, the weightmeasurement ratio g:ml of B material and solvent is 0.2-0.8:4-8.
Preferably, its raw material also comprises cyclodextrin, water and DMSO, and the volume ratio of water and DMSO is 2-12:1-10; The mixture of preferably water and DMSO and the envelope-bulk to weight ratio ml:g of B material are 4-8:0.2-0.8.
The invention also discloses the preparation method of the hyperbranched genophore of a kind of polyamide-amide, comprise the steps:
B material, C material, triethylamine and solvent are sent into reactor and carries out polyreaction, its temperature of reaction is 55-65 DEG C, and the reaction times is 30-40h, add A material and carry out ammonia solution, ammonia solution temperature is 55-65 DEG C, and the ammonia solution time is 34-52 hour, obtains the hyperbranched genophore of polyamide-amide.
The invention also discloses the preparation method of the hyperbranched genophore of a kind of polyamide-amide, comprise the steps:
Take 40-60wt%DMSO, add B material dissolution wherein and obtain the first solution completely, mixed by water with remaining DMSO, it is complete to add C material dissolution wherein, add cyclodextrin and obtain the second solution to saturated, first solution is dropped in the second solution, drip completely and add triethylamine, be warming up to 52-62 DEG C of reaction 32-38h, add A material and carry out ammonolysis reaction, ammonia solution temperature is 50-60 DEG C, and the ammonia solution time is 36 hours, and dialysis obtains the hyperbranched genophore of polyamide-amide; Preferably, the molecular weight of the dialysis membrane of dialysis use is 3000-14000.
Preferably, after ammonolysis reaction, also comprise concentrated, purifying; Preferably, concentration process carries out in Rotary Evaporators; In purge process, in the concentrated material obtained, add cold acetone and/or ether precipitates, filter, in the throw out obtained, add distilled water dissolve, continue to add cold acetone and/or ether precipitates, repeat 3-5 time successively.
The invention also discloses the hyperbranched genophore of a kind of polyamide-amide as the application of genophore for anticancer therapy.
The raw material of the hyperbranched genophore of polyamide-amide of the present invention is easy to get, and cost is low, and by the ratio between component each in conservative control raw material, generate the hyperbranched genophore of polyamide-amide containing polyamino, its degree of branching is better, there is the different degree of branching additionally by adopting single stage method preparation and there are the goods of identical repeating unit and similar molecular weight, preparation process is effective and method is simple, in concrete preparation process, first C material is utilized, B material carries out being obtained by reacting material under triethylamine catalysis good biocompatibility and lower cytotoxicity, secondly the hyperbranched genophore of polyamide-amide obtained after adding A material ammonia solution further has the 3-D solid structure being different from common genophore, more excellent vivo and vitro transfection efficiency can be obtained better with DNA compound, the cyclodextrin of further interpolation, effectively can control the branched structure of the hyperbranched genophore of polyamide-amide, when described goods are used as genophore, little to the toxicity of cell, surge capability is good, and have excellent DNA compound compressed ability, and raw material is easy to get, cost is lower, can be widely used in anticancer therapy, significantly reduces the cost of anticarcinogen, by popular colony is accepted.
Accompanying drawing explanation
Fig. 1 is the schematic arrangement of the side chain of the hyperbranched genophore of polyamide-amide that the present invention proposes;
Fig. 2 is the two-dimentional nuclear magnetic spectrogram of the hyperbranched genophore of polyamide-amide that the present invention proposes
1h-
1h-COSY;
Fig. 3 is the one dimension nuclear magnetic spectrogram of the hyperbranched genophore of polyamide-amide that the present invention proposes
13c-NMR;
Fig. 4 is the degree of branching of the different super branched molecules of the hyperbranched genophore of polyamide-amide that the present invention proposes;
Fig. 5 is the gel osmoticing chromatogram analysis table of the hyperbranched genophore of polyamide-amide that the present invention proposes;
Fig. 6 is Hydration fluid mechanics radius and the Zeta potential figure thereof of the hyperbranched genophore of polyamide-amide that the present invention proposes;
Fig. 7 is the infrared spectrum of the hyperbranched genophore of polyamide-amide that the present invention proposes;
Fig. 8 is the polyamide-amide gel electrophoresis figure after the ammonia solution that proposes of the present invention.
Embodiment
Below, by specific embodiment, technical scheme of the present invention is described in detail.
Embodiment 1
The hyperbranched genophore of a kind of polyamide-amide, its raw material comprises A material, B material, C material and triethylamine;
A material molecular formula is: C
2 (n-1)h
5n-2n
n, wherein n=2,3,4,5;
C material structure formula is:
wherein n=0,1,2,3,4,5,6, R=C
mh
2m+1, m=1,2,3,4,5;
B material structure formula is:
n=1,2,3,4,5,6; Or
x＝y＝1,2,3,4,5,6。
Embodiment 2
The hyperbranched genophore of a kind of polyamide-amide, its raw material comprises N, N '-methylene-bisacrylamide, Cys methyl ester hydrochloride, triethylamine, three second tetramine and water.
A material can be the polyethylene polyamine of 250-300 for quadrol, three second tetramines or molecular weight.
The mol ratio of N, N '-methylene-bisacrylamide and Cys methyl ester hydrochloride is the mol ratio 1:80 of 3:2.4, N, N '-methylene-bisacrylamide and three second tetramines.The weightmeasurement ratio g:ml of N, N '-methylene-bisacrylamide and aqueous solvent is 0.8:4.
Embodiment 3
The hyperbranched genophore of a kind of polyamide-amide, its raw material comprises N, and N '-methylene-bisacrylamide, Cys methyl ester hydrochloride, triethylamine, molecular weight are polyethylene polyamine and the solvent of 275.
The mol ratio of N, N '-methylene-bisacrylamide and Cys methyl ester hydrochloride is the mol ratio 1:120 of 2.5:2.9, N, N '-methylene-bisacrylamide and polyethylene polyamine.The weightmeasurement ratio g:ml of N, N '-methylene-bisacrylamide and solvent is 0.2:8.Solvent by volume part comprises 1 part, water, DMSO5 part.
Embodiment 4
The hyperbranched genophore of a kind of polyamide-amide, its raw material comprises N, N '-methylene-bisacrylamide, Cys methyl ester hydrochloride, triethylamine, quadrol, and solvent.
The mol ratio of N, N '-methylene-bisacrylamide and Cys methyl ester hydrochloride is the mol ratio 1:95 of 2:3, N, N '-methylene-bisacrylamide and quadrol.The weightmeasurement ratio g:ml of N, N '-methylene-bisacrylamide and solvent is 0.6:5.Solvent by volume part comprises 5 parts, water, DMSO1 part.
The preparation method of the hyperbranched genophore of described polyamide-amide, comprises the steps:
By N, N '-methylene-bisacrylamide, Cys methyl ester hydrochloride, triethylamine and solvent are sent into reactor and are carried out polyreaction, its temperature of reaction is 55 DEG C, reaction times is 40h, add quadrol ammonia solution, ammonia solution temperature is 55 DEG C, and the ammonia solution time is 52 hours, obtains the hyperbranched genophore of polyamide-amide.
Embodiment 5
The hyperbranched genophore of a kind of polyamide-amide, its raw material comprises N, N '-methylene-bisacrylamide, Cys methyl ester hydrochloride, triethylamine, three second tetramines, and solvent.
The mol ratio of N, N '-methylene-bisacrylamide and Cys methyl ester hydrochloride is the mol ratio 1:115 of 3:2.5, N, N '-methylene-bisacrylamide and three second tetramines.The weightmeasurement ratio g:ml of N, N '-methylene-bisacrylamide and solvent is 0.3:8.Solvent by volume part comprises 4 parts, water, DMSO2 part.
The preparation method of the hyperbranched genophore of described polyamide-amide, comprises the steps:
By N, N '-methylene-bisacrylamide, Cys methyl ester hydrochloride, triethylamine and solvent are sent into reactor and are carried out polyreaction, its temperature of reaction is 65 DEG C, reaction times is 30h, add three second tetramine ammonia solutions, ammonia solution temperature is 65 DEG C, and the ammonia solution time is 34 hours, obtains the hyperbranched genophore of polyamide-amide.
Embodiment 6
The hyperbranched genophore of a kind of polyamide-amide, its raw material comprises N, N '-methylene-bisacrylamide, Cys methyl ester hydrochloride, triethylamine, and molecular weight is 275 polyethylene polyamines, and solvent.
The mol ratio 1:100 of the mol ratio of N, N '-methylene-bisacrylamide and Cys methyl ester hydrochloride to be 3:2, N, N '-methylene-bisacrylamide and molecular weight be 275 polyethylene polyamines.The weightmeasurement ratio g:ml of N, N '-methylene-bisacrylamide and solvent is 0.45:6.Solvent by volume part comprises 3 parts, water, DMSO3 part.
The preparation method of the hyperbranched genophore of described polyamide-amide, comprises the steps:
By N, N '-methylene-bisacrylamide, Cys methyl ester hydrochloride, triethylamine and solvent are sent into reactor and are carried out polyreaction, its temperature of reaction is 60 DEG C, reaction times is 36h, adding molecular weight is 275 polyethylene polyamine ammonia solutions, ammonia solution temperature is 60 DEG C, and the ammonia solution time is 36 hours, obtains the hyperbranched genophore of polyamide-amide.
Embodiment 7
The hyperbranched genophore of a kind of polyamide-amide, its raw material comprises N, N '-methylene-bisacrylamide, Cys methyl ester hydrochloride, triethylamine, quadrol, water, DMSO, cyclodextrin.
The mol ratio of N, N '-methylene-bisacrylamide and Cys methyl ester hydrochloride is the mol ratio 1:119 of 4:1, N, N '-methylene-bisacrylamide and quadrol; Water and DMSO mixture and N, N '-methylene-bisacrylamide envelope-bulk to weight ratio ml:g are 7:0.8.The volume ratio of water and DMSO is 2:4.
The preparation method of the hyperbranched genophore of described polyamide-amide, comprises the steps:
Take 60wt%DMSO, add N wherein, N '-methylene-bisacrylamide, dissolve and obtain the first solution completely, water is mixed with remaining DMSO, adding Cys methyl ester hydrochloride wherein dissolves completely, add cyclodextrin and obtain the second solution to saturated, first solution is dropped in the second solution, drip completely and add triethylamine, be warming up to 62 DEG C of reaction 37h, add quadrol and carry out ammonolysis reaction, ammonia solution temperature is 58 DEG C, the ammonia solution time is 36 hours, concentrate in Rotary Evaporators, in the concentrated material obtained, add cold acetone precipitate, filter, in the throw out obtained, add distilled water dissolve, continue to add cold acetone to precipitate, repeat 3 times successively, select again molecular weight be 3000 dialysis membrane dialyse, vacuum lyophilization obtains the hyperbranched genophore of polyamide-amide for 24 hours.
Embodiment 8
The hyperbranched genophore of a kind of polyamide-amide, its raw material comprises N, N '-methylene-bisacrylamide, Cys methyl ester hydrochloride, triethylamine, three second tetramines, water, DMSO, and cyclodextrin.
The mol ratio of N, N '-methylene-bisacrylamide and Cys methyl ester hydrochloride is the mol ratio 1:82 of 2.1:1.4, N, N '-methylene-bisacrylamide and three second tetramines.Water and DMSO mixture and N, N '-methylene-bisacrylamide envelope-bulk to weight ratio ml:g are 4:0.26.The volume ratio of water and DMSO is 2:4.
The preparation method of the hyperbranched genophore of described polyamide-amide, comprises the steps:
Take 41wt%DMSO, add N wherein, N '-methylene-bisacrylamide, dissolve and obtain the first solution completely, water is mixed with remaining DMSO, adding Cys methyl ester hydrochloride wherein dissolves completely, add cyclodextrin and obtain the second solution to saturated, first solution is dropped in the second solution, drip completely and add triethylamine, be warming up to 53 DEG C of reaction 33h, add three second tetramines and carry out ammonolysis reaction, ammonia solution temperature is 52 DEG C, the ammonia solution time is 36 hours, concentrate in Rotary Evaporators, in the concentrated material obtained, add ether precipitate, filter, in the throw out obtained, add distilled water dissolve, continue to add ether to precipitate, repeat 4 times successively, select again molecular weight be 14000 dialysis membrane dialyse, vacuum lyophilization obtains the hyperbranched genophore of polyamide-amide for 24 hours.
Embodiment 9
The hyperbranched genophore of a kind of polyamide-amide, its raw material comprises N, N '-methylene-bisacrylamide, Cys methyl ester hydrochloride, triethylamine, and molecular weight is 275 polyethylene polyamines, water, DMSO, and cyclodextrin.
The mol ratio of N, N '-methylene-bisacrylamide and Cys methyl ester hydrochloride is the mol ratio 1:100 of 3:2, N, N '-methylene-bisacrylamide and polyethylene polyamine.Water and DMSO mixture and N, N '-methylene-bisacrylamide envelope-bulk to weight ratio ml:g are 6:0.45.The volume ratio of water and DMSO is 1:5.
The preparation method of the hyperbranched genophore of described polyamide-amide, comprises the steps:
Take 50wt%DMSO, add N wherein, N '-methylene-bisacrylamide, dissolve and obtain the first solution completely, water is mixed with remaining DMSO, adding Cys methyl ester hydrochloride wherein dissolves completely, add cyclodextrin and obtain the second solution to saturated, first solution is dropped in the second solution, drip completely and add triethylamine, be warming up to 60 DEG C of reaction 36h, add polyethylene polyamine and carry out ammonolysis reaction, ammonia solution temperature is 60 DEG C, the ammonia solution time is 36 hours, concentrate in Rotary Evaporators, cold acetone is added and ether precipitates in the concentrated material obtained, filter, in the throw out obtained, add distilled water dissolve, continue to add cold acetone and ether precipitates, repeat 5 times successively, select again molecular weight be 3500 dialysis membrane dialyse, vacuum lyophilization obtains the hyperbranched genophore of polyamide-amide for 24 hours.
The hyperbranched genophore of polyamide-amide described in embodiment 1-6 can be used as genophore for anticancer therapy.
Hyperbranched for described polyamide-amide genophore is carried out polymer characterization, wherein: a peacekeeping two-dimentional NMR spectrum uses the pulse sequence spectrograph record (deuterated water D2O-d2 is solvent) of Bruker500MHz standard; Adopt
1h-NMR;
13c-NMR;
13c,
1h-HMQC;
1h,
1h – COSY;
13c,
1the spectral characterization of H-HMBC.
By GPC (gel permeation chromatography) two detector, measure molecular weight.
Fourier transform infrared attenuated total reflectance attenuated total refraction spectrum (FTIR-ATR) carries out at the AVATAR370FT-IR type infrared spectrometer spectrograph of U.S. Buddhist nun high-tensile strength Company.
The hydrodynamic radius of MalvernZetasizer3000HS and computer for analysis software evaluation synthetic sample and Zeta potential, the sample dissolution of synthesis is in deionized water and at ultrasonic cell-break crusher machine and be evenly dispersed in deionized water.
As shown in Figure 1, Fig. 1 is the schematic arrangement of the side chain of the polyamide-amide that the present invention proposes.
As shown in Figure 2, Fig. 2 is the two-dimentional nuclear magnetic spectrogram of the polyamide-amide that the present invention proposes
1h-
1h-COSY.
As shown in Figure 3, Fig. 3 is the one dimension nuclear magnetic spectrogram of the polyamide-amide that the present invention proposes
13c-NMR.
As shown in Figure 4, Fig. 4 is the degree of branching of the different super branched molecules of the polyamide-amide that the present invention proposes.
As shown in Figure 5, Fig. 5 is the gel osmoticing chromatogram analysis table of the polyamide-amide that the present invention proposes.
As shown in Figure 6, Fig. 6 is Hydration fluid mechanics radius and the Zeta potential figure thereof of the polyamide-amide that the present invention proposes.
As shown in Figure 7, Fig. 7 is the infrared spectrum of the polyamide-amide that the present invention proposes.
As shown in Figure 8, Fig. 8 is the polyamide-amide gel electrophoresis figure after the ammonia solution that proposes of the present invention.
The above; be only the present invention's preferably embodiment; but protection scope of the present invention is not limited thereto; anyly be familiar with those skilled in the art in the technical scope that the present invention discloses; be equal to according to technical scheme of the present invention and inventive concept thereof and replace or change, all should be encompassed within protection scope of the present invention.
Claims (10)
1. the hyperbranched genophore of polyamide-amide, is characterized in that, its raw material comprises A material, B material, C material and triethylamine;
A material molecular formula is: C
2 (n-1)h
5n-2n
n, wherein n=2,3,4,5;
C material structure formula is:
wherein n=0,1,2,3,4,5,6, R=C
mh
2m+1, m=1,2,3,4,5;
B material structure formula is:
wherein n=1,2,3,4,5,6; Or
wherein x=y=1,2,3,4,5,6.
2. the hyperbranched genophore of polyamide-amide according to claim 1, it is characterized in that, A material is that one or more of the polyethylene polyamine of 250-300 form by quadrol, three second tetramines and molecular weight, B material is N, N '-methylene-bisacrylamide, C material is Cys methyl ester hydrochloride.
3. the hyperbranched genophore of polyamide-amide according to claim 2, it is characterized in that, the mol ratio of N, N '-methylene-bisacrylamide and Cys methyl ester hydrochloride is 2-4:1-3, the mol ratio 1:80-120 of N, N '-methylene-bisacrylamide and A material.
4. the hyperbranched genophore of the polyamide-amide according to any one of claim 1-3, is characterized in that, also comprise solvent, and solvent by volume part comprises water 1-6 part, DMSO0-5 part.
5. the hyperbranched genophore of polyamide-amide according to claim 4, is characterized in that, the weightmeasurement ratio g:ml of B material and solvent is 0.2-0.8:4-8.
6. the hyperbranched genophore of the polyamide-amide according to any one of claim 1-3, is characterized in that, its raw material also comprises cyclodextrin, water and DMSO, and the volume ratio of water and DMSO is 2-12:1-10; The mixture of preferably water and DMSO and the envelope-bulk to weight ratio ml:g of B material are 4-8:0.2-0.8.
7. a preparation method for the hyperbranched genophore of the polyamide-amide according to claim 1-5, is characterized in that, comprise the steps:
B material, C material, triethylamine and solvent are sent into reactor and carries out polyreaction, its temperature of reaction is 55-65 DEG C, and the reaction times is 30-40h, add A material and carry out ammonia solution, ammonia solution temperature is 55-65 DEG C, and the ammonia solution time is 34-52 hour, obtains the hyperbranched genophore of polyamide-amide.
8. a preparation method for the hyperbranched genophore of polyamide-amide according to claim 6, is characterized in that, comprise the steps:
Take 40-60wt%DMSO, add B material dissolution wherein and obtain the first solution completely, mixed by water with remaining DMSO, it is complete to add C material dissolution wherein, add cyclodextrin and obtain the second solution to saturated, first solution is dropped in the second solution, drip completely and add triethylamine, be warming up to 52-62 DEG C of reaction 32-38h, add A material and carry out ammonolysis reaction, ammonia solution temperature is 50-60 DEG C, and the ammonia solution time is 36 hours, and dialysis obtains the hyperbranched genophore of polyamide-amide; Preferably, the molecular weight of the dialysis membrane of dialysis use is 3000-14000.
9. the preparation method of the hyperbranched genophore of the polyamide-amide according to claim 7 or 8, is characterized in that, after ammonolysis reaction, also comprises concentrated, purifying; Preferably, concentration process carries out in Rotary Evaporators; In purge process, in the concentrated material obtained, add cold acetone and/or ether precipitates, filter, in the throw out obtained, add distilled water dissolve, continue to add cold acetone and/or ether precipitates, repeat 3-5 time successively.
10. the hyperbranched genophore of the polyamide-amide according to any one of claim 1-6 is used for the application of anticancer therapy as genophore.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101343359A (en) * | 2008-09-04 | 2009-01-14 | 上海交通大学 | Preparation method for aminophenol modified daiamid type tree shaped numerator |
| CN102140178A (en) * | 2010-12-28 | 2011-08-03 | 重庆工商大学 | Cyclodextrin-polyamidoamine cross-linked polymer and preparation method and application thereof |
| CN103110954A (en) * | 2013-01-31 | 2013-05-22 | 北京大学 | Cholesterol-modified biodegradable polycation carrier as well as preparation method and application thereof |
| CN104004196A (en) * | 2014-05-04 | 2014-08-27 | 健雄职业技术学院 | Preparation method and application of degradable hyperbranched polyamidoamine |
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2015
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101343359A (en) * | 2008-09-04 | 2009-01-14 | 上海交通大学 | Preparation method for aminophenol modified daiamid type tree shaped numerator |
| CN102140178A (en) * | 2010-12-28 | 2011-08-03 | 重庆工商大学 | Cyclodextrin-polyamidoamine cross-linked polymer and preparation method and application thereof |
| CN103110954A (en) * | 2013-01-31 | 2013-05-22 | 北京大学 | Cholesterol-modified biodegradable polycation carrier as well as preparation method and application thereof |
| CN104004196A (en) * | 2014-05-04 | 2014-08-27 | 健雄职业技术学院 | Preparation method and application of degradable hyperbranched polyamidoamine |
Non-Patent Citations (8)
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