Department of Pharmacology, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal 576104, Karnataka, India.
Department of Pharmaceutical Chemistry, Manipal College of Pharmaceutical Sciences, Manipal University, Manipal 576104, Karnataka, India.
Chem Biol Interact. 2015 May 25;233:81-94. doi: 10.1016/j.cbi.2015.03.015. Epub 2015 Mar 27.
The potential of cinnamic acid as an anti-inflammatory and anti-cancer agent has been studied previously. In our investigation, novel bio-isosters of cinnamyl sulfonamide hydroxamate were synthesized, characterized and confirmed for their structure and evaluated for cytotoxicity. Three NCEs namely, NMJ-1, -2 and -3 showed cell-growth inhibition in 6 human cancer cell lines with IC50 at the range of 3.3±0.15-44.9±2.6 μM. The hydroxamate derivatives of cinnamyl sulfonamide are reported inhibitors of HDAC enzyme. Thus, the effectiveness of these molecules was determined by whole cell HDAC assay in HCT 116 cell line. NMJ-2 (0.41±0.01 μM) exhibited better enzyme inhibition (IC50) compared to SAHA (2.63±0.07). In order to evaluate induction of apoptosis by treatment, Hoechst 33342 and AO/EB nuclear staining methods were used. Further, cell cycle analysis, Annexin V binding and caspase 3/7 activation assays were performed by flow cytometry where NMJ-2 significantly arrested the cell cycle at G2/M phase, increased Annexin V binding to the cell surface and activation of caspase-3/7. Bax/Bcl-2 ratio was observed by Western blot and showed an increase with NMJ-2 treatment. This was comparable to standard SAHA. The acute toxicity study (OECD-425) showed that NMJ-2 was safe up to 2000 mg/kg in rats. 1,2-Dimethyl hydrazine (DMH) was used to produce experimental colon adenocarcinoma in Wistar rats. 5-FU and NMJ-2 (100 mg/kg p.o. and 10 mg/kg i.p. once daily for 21 days, respectively) were administered to the respective groups. Both treatments significantly reduced ACFs, adenocarcinoma count, TNF-α, IL-6, nitrite and nitrate levels in colonic tissue. Our findings indicate that NMJ-2 has potent anti-cancer activity against colon cancer, by acting through HDAC enzyme inhibition and activation of intrinsic mitochondrial apoptotic pathway, with additional anti-inflammatory activity.
肉桂酸作为一种抗炎和抗癌药物的潜力此前已被研究过。在我们的研究中，合成了肉桂酰基磺酰胺羟肟酸的新型生物等排体，并对其结构进行了表征和确认，同时还对其细胞毒性进行了评估。三种新型化合物（NMJ-1、NMJ-2 和 NMJ-3）在 6 种人类癌细胞系中表现出细胞生长抑制作用，IC50 值在 3.3±0.15-44.9±2.6 μM 范围内。肉桂酰基磺酰胺的羟肟酸衍生物被报道为 HDAC 酶抑制剂。因此，通过全细胞 HDAC 测定法在 HCT 116 细胞系中测定这些分子的有效性。NMJ-2（0.41±0.01 μM）的酶抑制（IC50）优于 SAHA（2.63±0.07）。为了评估治疗引起的细胞凋亡，使用 Hoechst 33342 和 AO/EB 核染色法。此外，通过流式细胞术进行细胞周期分析、Annexin V 结合和 caspase 3/7 激活测定，NMJ-2 显著将细胞周期阻滞在 G2/M 期，增加了 Annexin V 与细胞表面的结合，并激活了 caspase-3/7。通过 Western blot 观察 Bax/Bcl-2 比值，发现 NMJ-2 处理后比值增加。这与标准 SAHA 相当。急性毒性研究（OECD-425）表明，NMJ-2 在大鼠中高达 2000 mg/kg 是安全的。1,2-二甲基肼（DMH）用于产生 Wistar 大鼠实验性结肠腺癌。5-FU 和 NMJ-2（分别以 100 mg/kg p.o.和 10 mg/kg i.p.每天一次，共 21 天）施用于相应组。两种治疗均显著降低了 ACFs、结肠腺癌计数、TNF-α、IL-6、结肠组织中的亚硝酸盐和硝酸盐水平。我们的研究结果表明，NMJ-2 通过抑制 HDAC 酶和激活内在线粒体凋亡途径发挥作用，具有潜在的抗癌活性，可有效治疗结肠癌，并具有额外的抗炎活性。
