Investigation of anti-leukemia molecular mechanism of ITR-284, a carboxamide analog, in leukemia cells and its effects in WEHI-3 leukemia mice.
ITR-284, a potent anti-leukemia agent of carboxamide derivative, has been shown to inhibit the proliferation of leukemia cells. In this study, the underlying molecular mechanisms in vitro and anti-leukemia activity in vivo of ITR-284 were investigated. ITR-284 reduced the cell viability and induced apoptosis in HL-60 and WEHI-3 leukemia cells. Following exposure of cells to 30 nM of ITR-284, there is a time-dependent decrease in the mitochondrial membrane potential (DeltaPsi(m)) and an increase in the reactive oxygen species (ROS). ITR-284 treatment also caused a time-dependent increase of Fas/CD95, cytosolic cytochrome c, cytosolic active form of caspase-8/-9/-3, cytosolic Apaf-1 and Bax, and the decrease of Bcl-2. However, the ITR-284-induced caspase-8/-9 and -3 activities can be blocked by pan-caspase inhibitor (Z-VAD-FMK). In addition, the anti-leukemia effects of ITR-284 in vivo were further evaluated in BALB/c mice inoculated with WEHI-3 cells. Orally treatment with ITR-284 (2 and 10mg/kg/alternate day for 7 times) increased the survival rate and prevented the loss of body weight in leukemia mice. The enlargement of spleen and infiltration of immature myeloblastic cells into spleen red pulp were significantly reduced in ITR-284-treated mice compared with control mice. Moreover, ITR-284 application can enhance the anti-leukemia effect of all-trans retinoic acid (ATRA). These results revealed that ITR-284 acted against both HL-60 and WEHI-3 in vitrovia both intrinsic and extrinsic apoptotic signaling pathways, and exhibited an anti-leukemic effect in a WEHI-3 orthotopic mice model of leukemia.
Wen
Yen-Fang
YF
Medicinal Chemistry Laboratory, Biomedical Engineering Research Laboratories, Industrial Technology Research Institute, Hsinchu 300, Taiwan.
Yang
Jai-Sing
JS
Kuo
Sheng-Chu
SC
Hwang
Chrong-Shiong
CS
Chung
Jing-Gung
JG
Wu
Hsi-Chin
HC
Huang
Wen-Wen
WW
Jhan
Jia-Hua
JH
Lin
Chung-Ming
CM
Chen
Hui-Jye
HJ
D000577:Amides / Q000737:chemistry / Q000627:therapeutic use; D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000627:therapeutic use*; D045744:Cell Line, Tumor; D006131:Growth Inhibitors / Q000737:chemistry / Q000627:therapeutic use*; D018922:HL-60 Cells; D006801:Humans; D007942:Leukemia, Experimental / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D013876:Thiophenes / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use*; D023041:Xenograft Model Antitumor Assays / Q000379:methods
D000577:Amides; D000970:Antineoplastic Agents; D006131:Growth Inhibitors; C545715:N-(2-dimethylaminoethyl)-4,8-dihydrobenzo(1,2-b,4,5-b')dithiophene-2-carboxamide; D013876:Thiophenes