Characterization Of Paeonol And Its Derivatives

The generation and evolution of hepatoma are related not only to abnormal differentiation and over-proliferation of hepatoma cells but also to apoptosis decrease. To develop the new drugs with inducing apoptosis of tumor cells will be a new strategy for curing cancer. Since arsenic trioxide has successfully used to cure acute promyelocytic leukemia(APL), arsenic compounds have then used to cure cancer since beginning of 1970s. From that, a series of arsenic compound are studied. Effect of arsenic compounds on cancer has paid increasing attention. Beyond that, paeonol is reported to be an effective against cancer in vivo and in vitro and be able to induce the apoptosis of tumor cells and affect the express of apoptotic genes. From these point of views, combining arsenic compounds with paeonol might be new drugs for anti-inflammatory, and its promotion of blood flow and anti-tumor. In this present, a series of derivatives of paeonol with arsenic compounds are synthesized, all of which are innovative compounds. The synthetic conditions of these compounds have been studied by orthogonal experiment and variance analysis. The optimization on extractive conditions of paeonol has been also studied. Anti-tumor activity of the innovative compounds on HepG2 cell line is screened, and the most effective and the optimal testing concentration are determined. Attempting to find out a kind of new organic arsenic drugs, simple structure and apoptosis induced to tumor cell in vivo and in vitro at a lower concentration, some methods are used by men of the growth inhibition assay, apoptotic analysis, and effective mechanism with HepG2 cell line.The main results and conclusions are as follows:1. The optimal extractive conditions of paeonolThe supercritical carbon dioxide is used to extraction of paeonol from Dabpi(a Chinese herb). The optimal extractive condition is obtained by orthogonal experiment, and the extractive effect, that is, extractive temperature is 45℃, the extractive pressure is 100Bar, and the extractive time is 75 minutes. The extraction rate is 94.1%.2. Eight synthesized derivatives of paeonol with arsenic compounds 2-hydroxy-4-methoxy-acetophenonesemi-p-aminobenzene(3a), 2-hydroxy-4-methoxy-acetophenonesemi-p-aminobenzenearsonic acid(3b), 2- methoxy-4-hydroxy-5- acetylazobenzene-4'-arsonic acid(4a), 1-o-hydroxy-p-methoxybenzoyl-2-p-aminobenzenearsonicacidethane(5a), 1-o-hydroxy-p- methoxybenzoyl-2-phenyl-2-p-aminobenzenearsonic acid-ethane(5b), 2'-hydroxy-4'-methoxybenzoyl-bromomethane(F1), 2'-hydroxy-4'-methoxy-5'-bromoacetophenone(F2), 2'-hydroxy-4'-methoxy-5'-bromobenzoyl- bromo-methane(F4-2), respectively.All of the eight compounds are innovative compounds, and the optimal synthetic conditions were as the following:● The yield of compound 3a (S1) reaches a maximum of 25.1% in such conditions as: (1) n (paeonol) :n (aniline)= 1:1; (2)The solution is normal butyl alcohol;(3) The refluxing time is 2 hours; (4)The refluxing temperature is 118℃.● The yield of compound 3b (S6) reaches a maximum of 28.4% in such conditions as:(1)The mol ratio between paeonol and arsanilic acid is 1:1 in normal butyl alcohol solution; (2)The refluxing time is 3 hours.● Changes of the mol ratio between paeonol and arsanilic acid have no effect on the yield of compound 4a (R2). The yield is 54.6% in these conditions: (1)The reaction temperature is controlled under 5℃; (2)The reaction time is 3 hours; (3)The pH value of reaction medium range from 8 to 10.● The optimal synthetic conditions of compound 5a (M3) are 1.5mL concentrated HCl/EtOH, the reaction temperature of 20℃, and the reaction time of 35 hours. The optimal synthetic conditions of compound 5b (M1) are 0.1-0.2mL concentrated HCl, the reaction temperature of 20℃, and the reaction time of 35 hours.● Paeonol bromide (F1, F2, F4-2): (1)compound F2 is synthesized in the condition of Br2/ absolute ether and a temperature of below zero;(2)Compound F1 is synthesized in the condition of absolute methanol or CHCl3/EtOAc and CuBr2; (3)Comp...