Macrocyclic polyamine compound based on dedpp-2tpa and its preparation method and application

Abstract

The invention discloses a macrocyclic polyamine compound based on DEDPP-2TPA, and a preparation method and use thereof. The disclosed compound is mainly prepared and synthesized by Suzuki coupling andcondensation reactions, an esterification reaction and a Click reaction, and the structure of the compound is also confirmed by nuclear magnetism and a mass spectrum; the compound has the propertiesof large Stock displacement and two-photon fluorescence, and an aggregation induction effect (AIE); after the compound interacts with nucleic acid, nanoparticles which are easy to uptake by cells areformed by co-aggregation; experiment of green fluorescent protein (GFP) and luciferase expression proves that a lipidosome formed by the compound with dioleoyl-phosphatidylethanolamine (DOPE) can be used as a non-viral gene vector; and cell uptake and release processes are traced by a single photon and two photons.

Description

technical field

[0001] The invention relates to a macrocyclic polyamine compound compound, in particular to a DEDPP-2TPA-based macrocyclic polyamine compound which can be used to prepare a transgene carrier and its preparation method and application. Background technique

[0002] Gene therapy is to introduce exogenous normal genes or genes with therapeutic functions into target cells to correct or compensate diseases caused by gene defects and abnormalities, so as to achieve therapeutic purposes. Gene therapy can provide new and powerful treatments for specific, chronic diseases, whether inherited or acquired. Due to the presence of phosphate groups in nucleic acids, nucleic acids are negatively charged, and there are many negatively charged proteins and glycolipid compounds on the cell membrane, which hinder the entry of nucleic acids into cells through endocytosis. Therefore, the smooth entry of nucleic acids into cells through gene carriers is a research key.

[0003] G...

Examples

Embodiment 1

[0052] A kind of macrocyclic polyamine compound based on DEDPP-2TPA, the structural formula (I) of said compound is as follows:

[0053]

[0054] In the formula (I), R is a macrocyclic polyamine [12]aneN 3 Structural unit, n is a positive integer.

[0055] Among them, when R is When n=4, it is recorded as compound 1;

[0056] when R is When n=6, it is recorded as compound 2;

[0057] when R is When n=8, it is recorded as compound 3;

[0058] Concrete synthetic route is as follows:

[0059]

[0060] Concrete synthetic steps are as follows:

[0061] 1), Compound 4 (1.1g, 3mmol), TPA-B(OH) 2 (2.17g, 7.5mmol), Pd(PPh 3 ) 4 (0.312g, 0.27mmol) into the two-necked bottle, nitrogen protection, inject anaerobic THF 100mL and KCO 3 Aqueous solution (2M, 50mL), reflux at 80°C for 8h, remove THF under reduced pressure, extract and dry with DCM, use petroleum ether / ethyl acetate (v / v=15:1) as eluent, and separate by column chromatography to obtain a yellow solid Compou...

Embodiment 2

[0095] Prepare solutions with different proportions (0-99%) of tetrahydrofuran and water, add compounds 1, 2 and 3 to prepare a solution (10 μM) with the same concentration, and measure the fluorescence emission intensity of compounds 1, 2 and 3 in different solvent ratios; The maximum value of the intensity and the change trend of the proportion of THF are plotted to obtain figure 1 A~1C, in figure 1 In A to 1C, the X-axis represents the concentration of tetrahydrofuran, and the Y-axis represents the fluorescence intensity. Depend on figure 1 It can be concluded that the macrocyclic polyamine compound based on DEDPP-2TPA of the present invention has excellent AIE performance.

Embodiment 3

[0097] Compounds 1, 2 and 3 were prepared at 10 μM in different solvents, and their UV absorption and fluorescence emission were tested, and the spectrum was processed to obtain figure 2 A~2F. Depend on figure 2 It can be concluded that the DEDPP-2TPA-based macrocyclic polyamine compound of the present invention has the characteristics of large Stoke shift, long-wave excitation and low self-luminescence.

technical field

[0001] The invention relates to a macrocyclic polyamine compound compound, in particular to a DEDPP-2TPA-based macrocyclic polyamine compound which can be used to prepare a transgene carrier and its preparation method and application. Background technique

[0002] Gene therapy is to introduce exogenous normal genes or genes with therapeutic functions into target cells to correct or compensate diseases caused by gene defects and abnormalities, so as to achieve therapeutic purposes. Gene therapy can provide new and powerful treatments for specific, chronic diseases, whether inherited or acquired. Due to the presence of phosphate groups in nucleic acids, nucleic acids are negatively charged, and there are many negatively charged proteins and glycolipid compounds on the cell membrane, which hinder the entry of nucleic acids into cells through endocytosis. Therefore, the smooth entry of nucleic acids into cells through gene carriers is a research key.

[0003] G...

Examples

Embodiment 1

[0052] A kind of macrocyclic polyamine compound based on DEDPP-2TPA, the structural formula (I) of said compound is as follows:

[0053]

[0054] In the formula (I), R is a macrocyclic polyamine [12]aneN 3 Structural unit, n is a positive integer.

[0055] Among them, when R is When n=4, it is recorded as compound 1;

[0056] when R is When n=6, it is recorded as compound 2;

[0057] when R is When n=8, it is recorded as compound 3;

[0058] Concrete synthetic route is as follows:

[0059]

[0060] Concrete synthetic steps are as follows:

[0061] 1), Compound 4 (1.1g, 3mmol), TPA-B(OH) 2 (2.17g, 7.5mmol), Pd(PPh 3 ) 4 (0.312g, 0.27mmol) into the two-necked bottle, nitrogen protection, inject anaerobic THF 100mL and KCO 3 Aqueous solution (2M, 50mL), reflux at 80°C for 8h, remove THF under reduced pressure, extract and dry with DCM, use petroleum ether / ethyl acetate (v / v=15:1) as eluent, and separate by column chromatography to obtain a yellow solid Compou...

Embodiment 2

[0095] Prepare solutions with different proportions (0-99%) of tetrahydrofuran and water, add compounds 1, 2 and 3 to prepare a solution (10 μM) with the same concentration, and measure the fluorescence emission intensity of compounds 1, 2 and 3 in different solvent ratios; The maximum value of the intensity and the change trend of the proportion of THF are plotted to obtain figure 1 A~1C, in figure 1 In A to 1C, the X-axis represents the concentration of tetrahydrofuran, and the Y-axis represents the fluorescence intensity. Depend on figure 1 It can be concluded that the macrocyclic polyamine compound based on DEDPP-2TPA of the present invention has excellent AIE performance.

Embodiment 3

[0097] Compounds 1, 2 and 3 were prepared at 10 μM in different solvents, and their UV absorption and fluorescence emission were tested, and the spectrum was processed to obtain figure 2 A~2F. Depend on figure 2 It can be concluded that the DEDPP-2TPA-based macrocyclic polyamine compound of the present invention has the characteristics of large Stoke shift, long-wave excitation and low self-luminescence.

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