Discovery of rafoxanide as a dual CDK4/6 inhibitor for the treatment of skin cancer

Shi,Xinan; Li,Hongjian; Shi,Anhua; Yao,Hong; Ke,Kunbin; Dong,Chao; Zhu,Ying; Qin,Yi; Ding,Ying; He,Yan Hong; Liu,Xu; Li,Ling; Lei,Ling; Hai,Qingshan; Chen,Wei; Leung,Kwong‑Sak; Wong,Man‑Hon; Kung,Hsiang‑Fu; Lin,Marie Chia‑Mi

doi:10.3892/or.2018.6533

Discovery of rafoxanide as a dual CDK4/6 inhibitor for the treatment of skin cancer

Authors:
- Xinan Shi
- Hongjian Li
- Anhua Shi
- Hong Yao
- Kunbin Ke
- Chao Dong
- Ying Zhu
- Yi Qin
- Ying Ding
- Yan Hong He
- Xu Liu
- Ling Li
- Ling Lei
- Qingshan Hai
- Wei Chen
- Kwong‑Sak Leung
- Man‑Hon Wong
- Hsiang‑Fu Kung
- Marie Chia‑Mi Lin
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Affiliations: Department of Medicine, Southwest Guizhou Vocational and Technical College for Nationalities, Xingyi, Guizhou 562400, P.R. China, Institute of Future Cities, The Chinese University of Hong Kong, Hong Kong 999077, P.R. China, Department of Physiology and Pathology, Yunnan University of Traditional Chinese Medicine, Kunming, Yunnan 650504, P.R. China, Jiangsu Cancer Biotherapy Institute, Xuzhou Medical College, Xuzhou, Jiangsu 221000, P.R. China, Biomedical Engineering Research Center, Kunming Medical University, Kunming, Yunnan 650504, P.R. China, Institute of Future Cities, The Chinese University of Hong Kong, Hong Kong 999077, P.R. China, Department of Computer Science and Engineering, The Chinese University of Hong Kong, Hong Kong 999077, P.R. China, Academy of Medical Sciences, Zhengzhou University, Zhengzhou, Henan 450000, P.R. China
Published online on: June 27, 2018 https://doi.org/10.3892/or.2018.6533
Pages: 1592-1600

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Abstract

Since cyclin‑dependent kinases 4/6 (CDK4/6) play pivotal roles in cell cycle regulation and are overexpressed in human skin cancers, CDK4/6 inhibitors are potentially effective drugs for skin cancer. In the present study, we present a mixed computational and experimental study attempting to repurpose approved small‑molecule drugs as dual CDK4/6 inhibitors for skin cancer treatment. We performed structure‑based virtual screening using the docking software idock, targeting an ensemble of CDK4/6 structures. We identified and selected nine compounds with significant predicted scores, and evaluated their cytotoxic effects in vitro in A375 and A431 human skin cancer cell lines. Rafoxanide was found to exhibit the highest cytotoxic effects (IC50: 1.09 µM for A375 and 1.31 µM for A431 cells). Consistent with the expected properties of CDK4/6 inhibitors, rafoxanide significantly increased the G1 phase population. Notably, we revealed that rafoxanide specifically decreased the expression of CDK4/6, cyclin D, retinoblastoma protein (Rb) and the phosphorylation of CDK4/6 and Rb. Furthermore, the anticancer effect of rafoxanide was demonstrated in vivo in BALB/C nude mice subcutaneously xenografted with human skin cancer A375 cells. Rafoxanide (40 mg/kg, i.p.) exhibited significant antitumor activity, comparable to that of oxaliplatin (5 mg/kg, i.p.). The combined administration of rafoxanide and oxaliplatin produced a synergistic therapeutic effect. To the best of our knowledge, the present study is the first to indicate that rafoxanide inhibits CDK4/6 activity and is a potential candidate drug for the treatment of human skin cancer.

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