Cationic lipid compounds and preparation method thereof
A technology of cationic lipids and compounds, applied in the field of medicine and biology, can solve the problems of toxicity and effectiveness restricting the application, liposomes cannot be widely used in research, toxicity and effectiveness hinder the application of cationic liposomes, etc., to increase delivery effects, effects that increase stability, change characteristics
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Embodiment 1
[0025] Example 1 The synthesis of trimethyl[2,3-(dilinoleyloxy)propyl]ammonium chloride
[0026]
[0027] 1) Add 13.35g of linoleyl alcohol (cis-9,12-octadecadienol), 7.78g of triethylamine, and 0.732g of 4-dimethylaminopyridine into a 0.25L three-necked flask, and add dichloromethane 30ml, stirred in -10℃ low-temperature circulation pump, lowered to -5℃, dissolved 13.35g of p-toluenesulfonyl chloride in 20ml of dichloromethane and added dropwise to the constant pressure dropping funnel, kept the liquid temperature at -5℃, 90mins After dropping, rinse the constant-pressure dropping funnel with 5ml of dichloromethane, slowly heat up, and react at room temperature for 15 hours. TLC petroleum ether: ethyl acetate 5:1 monitors the absence of linoleyl alcohol, separates the organic phase, and extracts the water phase with dichloromethane twice , the organic phase was dried with anhydrous magnesium sulfate, and the solution was evaporated by rotary evaporation to obtain a colorle...
Embodiment 2
[0033] Embodiment 2 siRNA preparation preparation
[0034] The trimethyl[2,3-(dilinoleyloxy)propyl]ammonium chloride synthesized in Example 1 was dissolved in ethanol, mixed with ALDH siRNA to produce a water-insoluble precipitate (1:1.5), After separating and drying the precipitate, dissolve the precipitate in chloroform or similar solvent, and further mix with other lipids, phosphatidylethanolamine:cholesterol:cholesterol-PEG (0.2:3:2.5:2) in chloroform, such as (WO / 2010 / 135207) described process. After removing the organic solvent, the dry preparation was hydrated with 9% sucrose water, ready for animal administration.
Embodiment 3
[0036] experiment method:
[0037] 1. All procedures used in in vivo animal studies were approved by the Institutional Animal Care and Use Committee (IACUC) and performed in accordance with local, state, and federal regulations. The formulation of siRNA prepared in Example 2 was injected by injecting 0.2 ml into the mouse tail vein. Harvested tissue and blood were analyzed for changes in gene expression. In addition, in order to illustrate the superiority of trimethyl[2,3-(dilinoleyloxy)propyl]ammonium chloride, trimethyl[2,3-(di-dodecyloxy)propyl base] ammonium chloride, trimethyl[2,3-(two-n-tetradecyloxy)propyl]ammonium chloride, trimethyl[2,3-(two-n-hexadecyloxy)propyl] Ammonium chloride, trimethyl[2,3-(dioleyloxy)propyl]ammonium chloride and other similar compounds were used as comparison, and the preparation method of the siRNA preparation was the same as in Example 2.
[0038]2. Isolation of mRNA: Two days after transfection, the cells were washed once with 100 μL of ...
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