Currently, several HDACi, including SAHA, LBH589, PXD101, MS-275, and FK228, are being examined in 5-BrdU clinical trials for their ability to treat various solid and hematological malignancies. The U.S. Food and Drug Administration recently approved SAHA and FK228 for the treatment of cutaneous T-cell lymphoma. HDAC inhibitors modulate the expression of several genes that regulate apoptosis, angiogenesis, cell cycle progression, and cellular differentiation. They have minimal toxicity against normal cells. Taken together, these findings are critical in designing target inhibitors of HDAC for the treatment of cancer and other diseases. A close view of these clinical trials with small molecule HDACi indicated the hydroxamic acid or N-hydroxyacrylamide group play an important role in HDAC activity. We previously reported identification of the indoline-1-sulfonamide-containing compounds with apparent anticancer activity. On the basis of the observations above, we designed and synthesized a series of new class of histone deacetylase inhibitors. HDACs are important targets for cancer therapy because of their ability to transcriptionally regulate the expression of genes that are involved in cell proliferation, differentiation, and apoptosis. HDACi are currently in clinical trials for the treatment of solid tumors and in leukemia patients. Two HDACi, SAHA and FK228, have received the US FDA approval for the treatment of patients with cutaneous T-cell lymphoma. In this study, we report the synthesis of a new chemical compound 3- -N-hydroxy-acrylamide, that has a potent HDACi activity. A 412997 dihydrochloride MPT0E028 was designed based on our prior research with microtubule destabilizing agents using indoline-1-arylsulfonamides as a core structure. After coupling an N-hydroxyacrylamide functional group at the 5- position of indoline ring, we afforded the desired MPT0E028. Furthermore, we evaluated the anti-cancer activity of MPT0E028 in vitro and in vivo. We found that MPT0E028 induced cytotoxicity in numerous human cancer cell lines from the NCI-60 panels and performed mechanistic studies in HCT116 cells, which showed high sensitivity to MPT0E028. When compared to SAHA, treatment with MPT0E028 induced stronger inhibition of cancer cell but not normal cell growth and produced a significantly higher number of sub-G1 cells as determined by flow cytometry. Also, MPT0E028 induced more profoundly caspase 3 and PARP activation. Taken together, MPT0E028 inhibits cancer cells growth and induces apoptotic cell-death. We demonstrated that MPT0E028 inhibits the activity of HDAC1, HDAC2 and HDAC8 in class I as well as HDAC6 in class IIb, but not HDAC4 in class IIa, and consistently induces acetylation of histone H3 and a-tubulin. Class I HDACs have been shown to be overexpressed in human colorectal cancer cells, which may contribute to perturbed cancer cell proliferation, differentiation, and apoptosis by both epigenetic or non-epigenetic modification. Inhibition of HDAC activity induced the intrinsic and extrinsic apoptotic pathway, leading to cancer cell death.