Experimental Study Of Guan-Fu Amino Alcohol Derivatives’ In Vitro Intervention On Two Digestive Tract Tumor Cell Lines Which Named AGS And SW480

Purpose:1. Compound Guan-Fu Amino Alcohol (GFAA) is chosen as the start molecule to modification and transformation. Addtionally, several new GFAA analogues has been designed and semisynthesized by acylation, which has enlarged the library of natural products.2. To study the synthetical products’antitumor activities on human gastric cancer cell lines AGS and human colon cancer cell lines SW480, to summary the structure-activity relationship, and to investigate the potential mechanisms.Methods:1. The structure of Guan-Fu Amino Alcohol (GFAA) is modified or altered by using various modern chromatographic techniques, and several of its analogues have been obtained via chemical design and synthesis by acylation. Their structures are confirmed by MS、 IR、1H NMR、13C NMR et al.2. Using MTT method to determine the IC50 of GFAA derivatives AA-Ac1、AA-Ac2、AA-Ac3、AA-Ac4、AA-Ac5、AA-Ac6 and AA-Ac7 on the proliferation index of the AGS and SW480 cells at 12h、24h and 48h.3. Using the flow cytometry analysis to detect the effects of GFAA derivatives on cell apoptosis and cell cycle of AGS and SW480 cells.Results:1. Guan-Fu Amino Alcohol’s 7 acylated derivatives are isolated for the firs structures are confirmed by MS、IR、1H NMR and 13C NMR, and their purity ar High Performance Liquid Chromatography (HPLC).2. AGS cells:5 (AA-Ac2、AA-Ac3、AA-Ac4、AA-Ac5、AA-Ac6) of the 7 acylated derivatives have inhibiting effect on the proliferation of AGS cells, the IC50 after 12 hours was: (20.95±0.30)μg/ml、(7.94±0.58)μg/ml、(7.39±0.61)μg/ml、>50μg/ml and (20.81±0.2)μg/ml respectively; the IC50 after 24 hours was:(14.12±0.11)μg/ml、(3.89±0.15)μg/ml、 (6.27±0.44)μg/ml、>50μg/ml、(16.09±0.19)μg/ml respectively; the IC50 after 48 hours was: (8.73±0.15)μg/ml、(3.09±0.24)μg/ml、(4.18±0.42)μg/ml、(19.83±0.07)μg/ml、(7.74±0.26)μg/ml.3. SW480 cells:4 (AA-Ac2、AA-Ac3、AA-Ac4、AA-Ac6) of the 7 acylated derivatives have inhibiting effect on the proliferation of SW480 cells, the IC50 after 12 hours was: (17.00±0.42)μg/ml、(7.48±0.52)μg/ml、(6.94±0.62)μg/ml、(34.84±0.20)μg/ml respectively; the IC50 after 24 hours was:(13.35±0.56)μg/ml、(4.50±0.32)μg/ml、(4.88±0.85)μg/ml、 (23.36±0.75)μg/ml respectively; the IC50 after 48 hours was:(10.38±026)μg/ml、 (3.60±0.20)μg/ml、(4.53±0.89)μg/ml、(11.91±0.64)μg/ml.4. Cell apoptosis analyzed by flow cytometry:compared with the control group, AGS cells treated with active compounds (AA-Ac2、AA-Ac3、AA-Ac4、AA-Ac5、AA-Ac6) after 48 hours, cell apoptosis rate is (13.25±0.78)%、(28.2±1.13)%、(26.75±1.77)%、(39.25±0.92)%、 (30.95±2.62)%、(40.85±0.35)% respectively; and SW480 cells treated with active compounds (AA-Ac2、AA-Ac3、AA-Ac4、AA-Ac6) after 48 hours, cell apoptosis rate is (4.45±1.06)%、 (22.85±1.63)%、(14.65±0.78)%、(18.05±0.78)%、(29.4±0.85)% respectively; compared with the control group, the active compounds group had a higher cell apoptosis, in which the late stage. The difference is statistically significant (P<0.05).5. Cell cycle analyzed by flow cytometry:to AGS cells, the G2/M stage of the cell cycle with active compounds (AA-Ac3、AA-Ac4、AA-Ac6) was increased to (22.07±1.34)%、 (16.08±0.76)% and (14.14±2.33)% compared to the control group (3.93±0.13)%, whereas AA-Ac2 and AA-Ac5 can increase the S stage of the cell cycle to (45.28±0.75)% and (45.46±0.55)%, compared to the control group (39.14±1.44)%; to SW480 cells, the G2/M stage of the cell cycle with active compounds (AA-Ac2、AA-Ac3、AA-Ac4、AA-Ac6) was increased to (60.04±1.34)%、(68.65±1.67)%、(63.42±0.84)% and (69.68±0.64)% compared to the control group (48.77±1.43)%. The difference is statistically significant (P<0.05).Conclusion:1. The technical route designed in this paper is reasonable and feasible. According to this route, we can really obtain Guan-Fu Amino Alcohol’s acylated derivatives.2. GFA A analogues (AA-Ac2、AA-Ac3、AA-Ac4、AA-Ac5、AA-Ac6) have inhibiting effect on the proliferation of AGS cells in time-dose dependence.3. GFAA analogues (AA-Ac2、AA-Ac3、AA-Ac4、AA-Ac6) have inhibiting effect on the proliferation of SW480 cells in time-dose dependence.4. GFA A analogues (A A-Ac2、AA-Ac3、AA-Ac4、AA-Ac5、AA-Ac6) can significantly induce apoptosis of AGS and SW480 cells, especially in the late stage.5. To AGS cells, GFA A analogues (AA-Ac3、AA-Ac4、AA-Ac6) can arrest the cell cycle in G2/M phase, whereas AA-Ac2 and AA-Ac5 can arrest the cell cycle in S phase; to SW480 cells, GFAA analogues (AA-Ac2、AA-Ac3、AA-Ac4、AA-Ac6) can arrest the cell cycle in G2/M phase.