CN103204999A - Synthetic method and application of water-soluble fluorescent dendrimers - Google Patents
Synthetic method and application of water-soluble fluorescent dendrimers Download PDFInfo
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Abstract
The invention discloses a synthetic method and application of water-soluble fluorescent dendrimers. Fluorescent dendrimers different in generation and carrying different functional groups are prepared from the kind of water-soluble fluorescent dendrimers by the aid of a 'divergent method' or 'convergence method'. The fluorescent dendrimers peripherally carrying ammonium salt cations have excellent water solubility and biocompatibility, are capable of entering living cells, can be combined with electronegative DNA (deoxyribonucleic acid), and can serve as genetic vectors to bring exogenous nucleic acid members DNA or RNA (ribonucleic acid) into the cells. Cellular uptake experiments show that along with increase of the generation, capabilities of penetrating cell membranes and carrying genes of the dendrimers are enhanced. The synthetic method and the application have the advantages of convenience in synthesis, high efficiency, product purification and simplicity and the like, and the synthetic dendrimers are safe, low in toxicity, excellent in water solubility and good in light stability, can serve as multifunctional cell fluorescent labeled molecules and gene vectors, and are applied to scientific researches and genetic diagnosis in the field of biological medicine.
Description
Technical field
The invention belongs to that dendritic macromole is synthetic, genophore and biomass cells marker field, particularly the perylene class fluorescence dendritic macromole of a class synthesizing biological degradable is applied to the genophore of non-viral system.
Background technology
Follow the development of advanced biotechnology therapy, gene importing system becomes more important.Genophore commonly used at present mainly contains virus type and non-virus type two big classes, and virus type gene transfection carrier has the limitation of safety problem and low gene transfection efficient, has limited their range of application.The non-viral gene vector that research and development have security is the hot research problem of technical field of biological material always.The more non-viral gene vector system of research mainly contains positively charged ion heteropolymer type carrier, biodegradable polymeric system, many composite liposomes body system, thermosensitive type polymeric system etc. at present.The cationic genophore of studying often has polymine (PEI), polylysine (PLL) and polyamide-amide type dendrimer (PAMAM), and wherein the applied research of dendrimer especially comes into one's own.
Perylene and derivative thereof have good light, heat, chemical stability, almost 100% fluorescence quantum yield (FQY), narrow fluorescence emission peak, can separate with cell background fluorescence (395-479nm) and excellent dyeing behavior, be widely used in organic electro-optic device, laser dyes and biological fluorescent labeling field.
In recent years, this compounds has been applied to biological field, as protein target mark and cell specific marker etc., yet poorly water-soluble has directly influenced it in the application in this field, therefore improves the water-soluble of this compounds and keeps high-fluorescence quantum yield significant in the aqueous solution.Carry out the functional group modification in the periphery of perylene and can make its maximum absorption band that very big red shift takes place, can better avoid the cell background fluorescence.
Dendritic macromole is owing to its accurate three-dimensional manometer molecular structure, and intramolecule exists a large amount of cavitys, carries a large amount of surface functional groups, can be widely used in biomedicine field, as pharmaceutical carrier and genophore etc.The dendritic macromole that synthesize at present De Yi perylene derivative and is fluorescent core all has good water-solubility, biocompatibility, lower cytotoxicity and the cell marking characteristic of excellence.Yet these macromole synthesis steps are loaded down with trivial details, need to the protection of middle product with separate protection, and protect/de-protected process tends to reaction not exclusively and with side reaction, thereby cause the dendritic macromole structure to have defective; Simultaneously, these synthetic De perylene derivative dendritic macromoles present electric neutrality, also do not report they to be applied to aspects such as pharmaceutical carrier and genophore.Thereby, invent a kind of method for preparing water-soluble De perylene analog derivative fluorescence dendritic macromole simply, efficiently, have very important significance at biomedicine field (as cell marking, pharmaceutical carrier, genophore etc.) for it.
Summary of the invention
The object of the present invention is to provide that a class light, heat, chemical stability are good, good water solubility, biodegradable, structure designability De perylene class fluorescence dendritic macromole, and it has good biocompatibility, cell marking.At first carry four primary amine functional group De perylene fluorescent core A0 by the synthetic method preparation of maturation, then by unsymmetrical monomer selectivity " click " chemical reaction, synthesize different algebraically, carry the fluorescence dendritic macromole of different functional groups.Fluorescence cation dendroid macromolecule B1, the B2 and the B3 that carry different amido functional groups are cultivated with active somatic cell respectively, study the ability of its permeates cell membranes; B1, B2 and B3 is compound with DNA respectively, cultivate with cell then, study the ability of its passenger gene.
A synthetic Lei Yi perylene derivative is the dendritic macromole of fluorescent core, comprise: (1) is carried functional group De perylene analog derivative and analogue thereof (the present invention is selected the naphthalene of Ppolynuclear aromatic hydrocarbon, pyrene perylene, the derivative of terylene and they is as fluorescent emission group) with unsymmetrical monomer AB(AB type unsymmetrical monomer be the methacryloxy ethyl propenoate, methacryloxy propyl acrylate or methacryloxy ethoxy ethoxy ethyl propenoate) reaction after, repeat to add successively unsymmetrical monomer mercaptoethylamine and AB, by selectivity " click " chemical reaction Bei perylene processed class fluorescence dendritic macromole between AB and the mercaptoethylamine.(the functionalized modification of 2) perylene fluorescence dendritic macromole end groups: the outermost end that hydroxyl, PEG chain functional group is incorporated into the fluorescence dendritic macromole.(3) water-soluble cationic fluorescence dendritic macromole is applied to genophore, research B1, B2, B3 permeates cell membranes and passenger gene ability.Its concrete preparation process is:
1. the design of dendritic macromole is synthetic
1.1 be raw material with commercial fluorescent chemicals Si Xiu perylene acid anhydride (4Br-PDA) at first, has a high-fluorescence quantum yield De perylene analog derivative 4Br-PBI through amidate action is synthetic; Utilize substitution reaction with amino Yin Ru " island " position Dao perylene nuclear then, thereby obtain carrying four primary amine group De perylene compounds A0(compound A-40 Shu Yu perylene class fluorescent emission group series), its structural formula is as follows:
1.2 compound A-40 and methacryloxy ethyl propenoate (MAEA) are added in the reaction tubes, mol ratio is between 1:16-1:20, stirred overnight at room temperature under nitrogen atmosphere, at 48-53 ℃ of reaction 48-56h, obtain the first-generation fluorescence dendritic macromole (A1) of terminal methyl acrylate then;
1.3 above-mentioned A1 and mercaptoethylamine are dissolved between 1:9-1:11 in the dimethyl sulfoxide (DMSO) (DMSO) according to mol ratio, under nitrogen atmosphere, behind the stirring at room 30-45min, obtain the first-generation fluorescence dendritic macromole (B1) of aminofunctional;
1.4 above-claimed cpd B1 and MAEA are mixed between 1:32-1:80 in the adding reaction tubes according to mol ratio, stirred overnight at room temperature under nitrogen atmosphere, at 48-53 ℃ of reaction 56-72h, obtain the s-generation fluorescence dendritic macromole (A2) of terminal methyl acrylate then;
1.5 above-mentioned A2 and mercaptoethylamine are dissolved between 1:18-1:21 in the dimethyl sulfoxide (DMSO) (DMSO) according to mol ratio, under nitrogen atmosphere, behind the stirring at room 30-45min, obtain the s-generation fluorescence dendritic macromole (B2) of aminofunctional;
1.6 above-claimed cpd B2 and MAEA are mixed between 1:80-1:400 in the adding reaction tubes according to mol ratio, stirred overnight at room temperature under nitrogen atmosphere, at 48-53 ℃ of reaction 72-120h, obtain the third generation fluorescence dendritic macromole (A3) of terminal methyl acrylate then;
1.7 above-mentioned A3 and mercaptoethylamine are dissolved between 1:38-1:42 in the dimethyl sulfoxide (DMSO) (DMSO) according to mol ratio, under nitrogen atmosphere, behind the stirring at room 30-45min, obtain the third generation fluorescence dendritic macromole (B3) of aminofunctional;
The functional modification of 2 perylene class fluorescence dendritic macromole end groups
2.1 hydroxyl functional De perylene class fluorescence dendritic macromole is synthetic
Above-mentioned A1 and mercapto glycerol are dissolved among the DMSO between 1:18-1:22 according to mol ratio, and are catalyzer with the triethylamine, behind the stirring at room 22-26h, obtain the first-generation fluorescence dendritic macromole (C1) of hydroxy-functional under the nitrogen atmosphere;
Above-mentioned A2 and mercapto glycerol are dissolved among the DMSO between 1:36-1:42 according to mol ratio, and are catalyzer with the triethylamine, behind the stirring at room 35-38h, obtain the s-generation fluorescence dendritic macromole (C2) of hydroxy-functional under the nitrogen atmosphere;
Above-mentioned A3 and mercapto glycerol are dissolved among the DMSO between 1:80-1:100 according to mol ratio, and are catalyzer with the triethylamine, behind the stirring at room 38-48h, obtain the third generation fluorescence dendritic macromole (C3) of hydroxy-functional under the nitrogen atmosphere.
2.2 terminal PEG chain functionalization De perylene class fluorescence dendritic macromole is synthetic
Carry perylene compounds A0 and ethoxy ethoxy ethyl propylene acid esters stirred overnight at room temperature under solvent-free, catalyst-free, nitrogen atmosphere of four primary amine groups, at 48-52 ℃ of reaction 48-56h, synthesized the Yi Dai perylene class fluorescence dendritic macromole D1 of terminal PEG chain then; Compound B-11 and ethoxy ethoxy ethyl propylene acid esters stirred overnight at room temperature under solvent-free, catalyst-free, nitrogen atmosphere then at 48-55 ℃ of reaction 68-72h, have synthesized the 2nd generation perylene class fluorescence dendritic macromole D2 of terminal PEG chain;
3 water-soluble cationic fluorescence dendritic macromoles are applied to genophore
The fluorescence dendritic macromole that above-mentioned synthetic method is synthesized (B1, B2 B3) are applied to genophore, it is characterized in that:
With the synthetic first-generation to third generation fluorescence dendritic macromole (B1, B2, B3) be the acid treatment of 1-4M with concentration after, cultivate with active somatic cell, discovery B1, B2, B3 can both enter viable cell fast, shows excellent biological compatibility; This compounds has very strong ability of carrying gene simultaneously, and wherein the passenger gene ability of B3 is the highest.
The present invention has following beneficial effect:
1. by the new fluorescent core molecule of design, Shou Zai perylene " island " position has synthesized water miscible, the Ke biological degradation De perylene class fluorescence dendritic macromole of different algebraically by the method for " click " chemistry.Such macromole also has characteristics such as good light, heat, chemical stability, structure designability, excellent biocompatibility, cell marking.
2. the present invention utilizes unsymmetrical monomer that the method that a points of proximity hits chemical reaction is reacted, the goal tree dendritic macromolecules is unique product, reduced the synthetic and purification step of dendritic macromole, accelerate resultant velocity, improved quality and the Product Safety of fluorescence dendritic macromole.Simultaneously, the inventive method can be controlled according to actual needs and repeat to replace the unsymmetrical monomer AB of adding and the number of times of mercaptoethylamine, prepares the fluorescence dendritic macromole of different algebraically, as preparing 1-6 for the fluorescence dendritic macromole of different molecular weight.
3. the fluorescence dendritic macromole for preparing of the inventive method has good biocompatibility, and a large amount of amido functional groups is rich on the fluorescence cation dendroid macromolecule surface of carrying different amido functional groups, the amino protonated dendritic macromole that can make of end has positive charge, thereby can with electronegativity nucleic acid substances DNA or RNA interaction of molecules, can exogenous nucleic acid transhipment be entered in the cell as genophore.Experiment showed, that synthetic fluorescence dendritic macromole increases along with the increase of algebraically the efficient that cell carries out transgenosis, and cytotoxicity is very low, has good using value.
4. in addition, this synthetic route can be saved the synthetic and high needed plenty of time of algebraically dendritic macromole of separation and purification in the past, improves combined coefficient.
Description of drawings
The structural formula of figure 1 perylene analog derivative and analogue thereof.
The synthetic reacting flow chart that carries four primary amine group De perylene compounds A0 among Fig. 2 embodiment 1.
The preparation first-generation is to the reaction principle synoptic diagram of third generation fluorescence dendritic macromole among Fig. 3 embodiment 1.
The structural formula of hydroxyl functional De perylene class fluorescence dendritic macromole C1, C2, C3 among Fig. 4 embodiment 4.
The structural formula of Fig. 5 embodiment 5 middle-end PEG chain functionalization De perylene class fluorescence dendritic macromole D1, D2, D3.
The fluorescence emission spectrum spectrogram of dendritic macromole B1, B2, B3 among Fig. 6 embodiment 1.
Dendritic macromole B1, B2, B3 enter fluorescence imaging figure behind the active somatic cell among Fig. 7 embodiment 6.Fig. 9 A is the fluorescence imaging figure of B1 after active somatic cell is cultivated 24h; Fig. 9 B is the fluorescence imaging figure of B2 after active somatic cell is cultivated 12h; Fig. 9 C is the fluorescence imaging figure of B3 after active somatic cell is cultivated 12h.
Dendritic macromole B1, B2, B3 gene transfection test among Fig. 8 embodiment 7.2 μ M B1/100 μ M DNA mixtures, 2 μ M B2/100 μ M DNA mixtures, 2 μ M B3/100 μ M DNA mixtures (N/P=8:1) penetrate into the fluorescence imaging figure after active somatic cell is cultivated 48h.Figure A1 is the fluorescence imaging of dendritic macromole B1; Figure A2 is the fluorescence imaging of dendritic macromole B2; Figure A3 is the fluorescence imaging of dendritic macromole B3.Figure B1, figure B2, figure B3 all with CXR with reference to dye marker DNA.Figure C1 merges image for scheming A1 and scheming B1, and figure C2 is that figure A2 merges image with figure B2, schemes C3 for scheming A3 and figure B3 merging image.
Dendritic macromole B1 and DNA composite fluorescence image among Fig. 9 embodiment 7.
Embodiment
Embodiment 1 utilizes the click chemistry reaction method to synthesize the fluorescence dendritic macromole
(1) first-generation fluorescence dendritic macromole (A1) of synthetic terminal methyl acrylate
With 0.16g(0.16mmol) carry four primary amine group De perylene compounds A0 and 0.2g(3.15mmol) MAEA adds in two mouthfuls of reaction tubess of 10mL, stirred overnight at room temperature under nitrogen atmosphere, then at 50 ℃ of reaction 48h, after dropping to room temperature etc. temperature of reaction, with normal hexane washing reaction liquid 3 times, each 30mL normal hexane, vacuum-drying gets red oily product A 1, and productive rate is 98%.
1H-NMR(400Hz,CDCl
3):δppm:8.20(s,4H),7.09(d,8H),6.91(d,8H),6.11(s,8H),5.56(d,8H),4.32(br,32H),4.12(t,4H),2.87(t,16H),2.69(m,16H),2.51(t,16H),1.9(s,24H),1.59(m,4H),1.36(m,4H),0.90(t,6H).MS(MALDI-TOF,m/z)Calc.for?C
136H
158N
6O
40,2516.73;found:2515.8.
(2) the functionalized first-generation fluorescence dendritic macromole (B1) of synthesizing amino
Above-mentioned 0.201g (0.08mmol) A1 and 0.062g (0.81mmol) mercaptoethylamine are dissolved in the 1mL dimethyl sulfoxide (DMSO) (DMSO), under nitrogen atmosphere behind the stirring at room 45min, add 20mL methylene dichloride diluting soln, then with saturated icy salt solution washing four times, each 30mL icy salt solution, behind the extraction separatory, remove organic solvent through underpressure distillation and get red oily product B 1, productive rate is 98%.
1H-NMR(CDCl
3,400Hz):δppm:8.12(s,4H),7.03(d,8H),6.84(d,8H),4.22(m,32H),4.15(t,4H),2.81(t,16H),2.77(m,32H),2.61(m,24H),2.53(m,16H),2.43(t,16H),1.56-1.44(m,8H),1.18(s,24H),0.91(t,6H).
(3) s-generation fluorescence dendritic macromole (A2) of synthetic terminal methyl acrylate
With 0.228g(0.072mmol) compound B-11 and 0.5g(2.71mmol) MAEA adds in two mouthfuls of reaction tubess of 10mL, stirring at room 24h under nitrogen atmosphere, then at 50 ℃ of reaction 72h, after dropping to room temperature etc. temperature of reaction, with normal hexane washing reaction liquid 4 times, each 30mL normal hexane dissolves crude product again with methylene dichloride then, remove organic solvent through underpressure distillation and get red oily product A 2, productive rate is 90%.
1H-NMR(400Hz,CDCl
3):δppm:8.18(s,4H),7.10(d,8H),6.92(d,8H),6.12(s,16H),5.59(d,16H),4.33-4.27(br,96H),4.15(t,4H),2.90-2.42(m,168H),1.94(s,48H),1.55-1.48(m,8H)1.25(d,24H),0.89(t,6H).
(4) the functionalized s-generation fluorescence dendritic macromole (B2) of synthesizing amino
Above-mentioned 0.0725g (0.012mmol) A2 and 0.0195g (0.25mmol) mercaptoethylamine are dissolved in the 1mL dimethyl sulfoxide (DMSO) (DMSO), under nitrogen atmosphere behind the stirring at room 45min, add 20mL methylene dichloride diluting soln, then with saturated icy salt solution washing four times, each 30mL icy salt solution, behind the extraction separatory, remove organic solvent through underpressure distillation and get red oily product B 2, productive rate is 92%.
(5) synthetic third generation fluorescence dendritic macromole (B3)
With 0.087g(0.012mmol) compd B 2 and 0.5g(2.71mmol) MAEA adds in two mouthfuls of reaction tubess of 10mL, stirring at room 24h under nitrogen atmosphere, then at 50 ℃ of reaction 72h, after dropping to room temperature etc. temperature of reaction, with normal hexane washing reaction liquid 4 times, each 30mL normal hexane dissolves crude product again with methylene dichloride then, remove organic solvent through underpressure distillation and get red oily product A 3, productive rate is 95%.
1H-NMR(400MHz,CDCl
3):δppm:8.18(s,4H),7.11(d,J=7.1Hz,8H),6.92(s,8H),6.13(s,32H),5.60(s,32H),4.31(d,J=22.1Hz,224H),4.14(t,4H),2.88-2.44(m,408H),1.95(s,96H),1.50-1.42(m,8H)1.25(d,J=6.5Hz,72H),0.88(t,6H).
Above-mentioned 0.074g (0.006mmol) A3 and 0.0195g (0.25mmol) mercaptoethylamine are dissolved in the 1mL dimethyl sulfoxide (DMSO) (DMSO), under nitrogen atmosphere behind the stirring at room 45min, add 20mL methylene dichloride diluting soln, then with saturated icy salt solution washing four times, each 30mL icy salt solution, behind the extraction separatory, remove organic solvent through underpressure distillation and get red oily product B 3, productive rate is 90%.
Except adopting the methacryloxy propyl acrylate to replace the methacryloxy ethyl propenoate and carry four primary amine group De perylene compounds A0 reactions in the step (1), other operation all is same as embodiment 1, prepares the first-generation to San Dai perylene class fluorescence dendritic macromole.
Embodiment 3
Except adopting methacryloxy ethoxy ethoxy ethyl propenoate to replace the methacryloxy ethyl propenoate and carry four primary amine group De perylene compounds A0 reactions in the step (1), other operation all is same as embodiment 1, prepares the first-generation to San Dai perylene class fluorescence dendritic macromole.
Synthesizing of embodiment 4 hydroxyl functional De perylene class fluorescence dendritic macromoles
(1) according to embodiment 1 synthetic compound A1, A2, A3;
(2) compd A 1 is solvent with mercapto glycerol at DMSO, and triethylamine is under the catalyzer condition behind the room temperature reaction 24h, can obtain the Yi Dai perylene class fluorescence dendritic macromole C1 of hydroxyl functionalization;
(3) compd A 2 is solvent with mercapto glycerol at DMSO, and triethylamine is under the catalyzer condition behind the room temperature reaction 36h, can obtain the Er Dai perylene class fluorescence dendritic macromole C2 of hydroxyl functionalization;
(4) compound A-13 and mercapto glycerol are solvent at DMSO, and triethylamine is under the catalyzer condition behind the room temperature reaction 40h, can obtain the San Dai perylene class fluorescence dendritic macromole C3 of hydroxyl functionalization.
Synthesizing of embodiment 5 end PEG chain functionalization De perylene class fluorescence dendritic macromoles
(1) carries perylene compounds A0 and ethoxy ethoxy ethyl propylene acid esters stirred overnight at room temperature under solvent-free, catalyst-free, nitrogen atmosphere of four primary amine groups, at 50 ℃ of reaction 48h, synthesized the Yi Dai perylene class fluorescence dendritic macromole D1 of end PEG chain then;
(2) according to embodiment 1 synthetic compound B1, B2;
(3) compound B-11 and ethoxy ethoxy ethyl propylene acid esters stirred overnight at room temperature under solvent-free, catalyst-free, nitrogen atmosphere then at 50 ℃ of reaction 72h, have synthesized the 2nd generation perylene class fluorescence dendritic macromole D2 of end PEG chain;
(4) compd B 2 and ethoxy ethoxy ethyl propylene acid esters stirred overnight at room temperature under solvent-free, catalyst-free, nitrogen atmosphere then at 50 ℃ of reaction 80h, have synthesized the 3rd generation perylene class fluorescence dendritic macromole D3 of end PEG chain.
Embodiment 6 active somatic cells picked-up experiment
With the synthetic first-generation to third generation fluorescence dendritic macromole (B1, B2 B3) with after the HBr acidification of 2M, cultivate with active somatic cell, find B1, B2, B3 can both enter viable cell, shows excellent biological compatibility.
Embodiment 7 gene transfections experiment
Utilize dendritic macromole B1 of the present invention, B2, B3 bonding DNA transfection active somatic cell has been investigated the compound ratio of DNA and dendritic macromole, and namely the terminal ammonium salt number of dendritic macromole and the number of dna molecular phosphate radical are than (N/P) influence to transfection experiment.By research with different N/P than (1:1,2:1,4:1,8:1) DNA of Xing Chenging and B1, B2, found that of B3 mixture transfectional cell, B3 at all N/P than all showing very high transfection activity under the condition.When comparing, N/P is 8:1,2 μ M B3, and during 100 μ M DNA, the transfection efficiency of the active somatic cell of B3 is best.
Claims (8)
1. the synthetic method of water soluble fluorescence dendritic macromole is characterized in that:
Four primary amine group De perylene compounds, hereinafter to be referred as compound A-40, the structural formula of compound A-40 is as follows:
Compound A-40 and methacryloxy ethyl propenoate MAEA are added in the reaction tubes, mol ratio is between 1:16-1:20, stirred overnight at room temperature under nitrogen atmosphere then at 48-53 ℃ of reaction 48-56h, obtains the first-generation fluorescence dendritic macromole A1 of terminal methyl acrylate.
2. the synthetic method of water soluble fluorescence dendritic macromole according to claim 1 is characterized in that also comprising:
Above-mentioned A1 and mercaptoethylamine are dissolved between 1:9-1:11 among the dimethyl sulfoxide (DMSO) DMSO according to mol ratio, under nitrogen atmosphere, behind the stirring at room 30-45min, obtain the first-generation fluorescence dendritic macromole B1 of aminofunctional.
3. the synthetic method of water soluble fluorescence dendritic macromole according to claim 2 is characterized in that also comprising:
Above-claimed cpd B1 and MAEA are mixed between 1:32-1:80 in the adding reaction tubes according to mol ratio, and stirred overnight at room temperature under nitrogen atmosphere then at 48-53 ℃ of reaction 56-72h, obtains the s-generation fluorescence dendritic macromole A2 of terminal methyl acrylate.
4. the synthetic method of water soluble fluorescence dendritic macromole according to claim 3 is characterized in that also comprising:
Above-mentioned A2 and mercaptoethylamine are dissolved between 1:18-1:21 among the dimethyl sulfoxide (DMSO) DMSO according to mol ratio, under nitrogen atmosphere, behind the stirring at room 30-45min, obtain the s-generation fluorescence dendritic macromole B2 of aminofunctional.
5. the synthetic method of water soluble fluorescence dendritic macromole according to claim 4 is characterized in that also comprising:
Above-claimed cpd B2 and MAEA are mixed between 1:80-1:400 in the adding reaction tubes according to mol ratio, stirred overnight at room temperature under nitrogen atmosphere, at 48-53 ℃ of reaction 72-120h, obtain the third generation fluorescence dendritic macromole A3 of terminal methyl acrylate then;
Above-mentioned A3 and mercaptoethylamine are dissolved between 1:38-1:42 among the dimethyl sulfoxide (DMSO) DMSO according to mol ratio, under nitrogen atmosphere, behind the stirring at room 30-45min, obtain the third generation fluorescence dendritic macromole B3 of aminofunctional.
6. according to the synthetic method of claim 1 or 3 or 5 described water soluble fluorescence dendritic macromoles, it is characterized in that also comprising:
Above-mentioned A1 and mercapto glycerol are dissolved among the DMSO between 1:18-1:22 according to mol ratio, and are catalyzer with the triethylamine, behind the stirring at room 22-26h, obtain the first-generation fluorescence dendritic macromole C1 of hydroxy-functional under the nitrogen atmosphere;
Above-mentioned A2 and mercapto glycerol are dissolved among the DMSO between 1:36-1:42 according to mol ratio, and are catalyzer with the triethylamine, behind the stirring at room 35-38h, obtain the s-generation fluorescence dendritic macromole C2 of hydroxy-functional under the nitrogen atmosphere;
Above-mentioned A3 and mercapto glycerol are dissolved among the DMSO between 1:80-1:100 according to mol ratio, and are catalyzer with the triethylamine, behind the stirring at room 38-48h, obtain the third generation fluorescence dendritic macromole C3 of hydroxy-functional under the nitrogen atmosphere.
7. according to the synthetic method of claim 2 or 4 described water soluble fluorescence dendritic macromoles, it is characterized in that also comprising:
Compound A-40 and ethoxy ethoxy ethyl propylene acid esters stirred overnight at room temperature under solvent-free, catalyst-free, nitrogen atmosphere then at 48-52 ℃ of reaction 48-56h, have synthesized a generation perylene class fluorescence dendritic macromole D1 of end PEG chain;
Compound B-11 and ethoxy ethoxy ethyl propylene acid esters stirred overnight at room temperature under solvent-free, catalyst-free, nitrogen atmosphere then at 48-55 ℃ of reaction 68-72h, have synthesized the 2nd generation perylene class fluorescence dendritic macromole D2 of end PEG chain;
Compd B 2 and ethoxy ethoxy ethyl propylene acid esters stirred overnight at room temperature under solvent-free, catalyst-free, nitrogen atmosphere then at 48-60 ℃ of reaction 78-100h, have synthesized the 3rd generation perylene class fluorescence dendritic macromole D3 of end PEG chain.
8. adopt the synthetic macromolecular application of claim 2 or 4 or 5 described methods, it is characterized in that also comprising:
With synthetic dendritic macromole B1, B2, B3 be with after the sour acidification of 1-4M, by adding in the cell culture fluid, cultivate active somatic cell more than 1 hour after, these molecules can both enter active somatic cell; Carrier is served as in these molecular energies and exogenous nucleic acid matter DNA or RNA combination, carries exogenous nucleic acid material transfection active somatic cell.
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| CN103999853A (en) * | 2014-06-11 | 2014-08-27 | 中国农业大学 | Application of fluorescence arborization nanometer macromolecule in preparing drug carrier |
| CN107033267A (en) * | 2017-03-15 | 2017-08-11 | 北京化工大学 | A series of preparation of water-soluble polymers based on Lay quinoline based compound and its as photothermal reagent in biomedical application |
| CN107189075A (en) * | 2017-06-06 | 2017-09-22 | 东北石油大学 | A kind of dendrimer compound and its preparation method and use |
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| CN103539951A (en) * | 2013-09-29 | 2014-01-29 | 北京化工大学 | Water-soluble hyperbranched fluorescent polymer and preparation method and applications thereof |
| CN103539951B (en) * | 2013-09-29 | 2015-10-14 | 北京化工大学 | A kind of water-soluble ultrabranching fluorescent polymer and preparation method thereof and application |
| CN103755703A (en) * | 2013-12-24 | 2014-04-30 | 北京化工大学 | Synthesis and application of double-function water-soluble perylene bisimide derivative |
| CN103755703B (en) * | 2013-12-24 | 2016-05-18 | 北京化工大学 | A kind of synthetic and application of difunctional water-soluble perylene diimide derivatives |
| CN103999853A (en) * | 2014-06-11 | 2014-08-27 | 中国农业大学 | Application of fluorescence arborization nanometer macromolecule in preparing drug carrier |
| CN107033267A (en) * | 2017-03-15 | 2017-08-11 | 北京化工大学 | A series of preparation of water-soluble polymers based on Lay quinoline based compound and its as photothermal reagent in biomedical application |
| CN107033267B (en) * | 2017-03-15 | 2020-04-28 | 北京化工大学 | Preparation of a series of water-soluble polymers based on Lerinyl compounds and application of water-soluble polymers as photothermal reagents in biomedicine |
| CN107189075A (en) * | 2017-06-06 | 2017-09-22 | 东北石油大学 | A kind of dendrimer compound and its preparation method and use |
| CN107189075B (en) * | 2017-06-06 | 2018-03-02 | 东北石油大学 | A kind of dendrimer compound and its preparation method and use |
| CN111704716A (en) * | 2020-07-03 | 2020-09-25 | 浙江大学 | Fluorescent polylysine dendrimer, and preparation method and application thereof |
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