Downregulation of PD‑L1 via amide analogues of brefelamide: Alternatives to antibody‑based cancer immunotherapy

Zhang,Jing; Yamada,Osamu; Kida,Shinya; Murase,Shinya; Hattori,Toshio; Oshima,Yoshiteru; Kikuchi,Haruhisa

doi:10.3892/etm.2020.8553

Downregulation of PD‑L1 via amide analogues of brefelamide: Alternatives to antibody‑based cancer immunotherapy

Authors:
- Jing Zhang
- Osamu Yamada
- Shinya Kida
- Shinya Murase
- Toshio Hattori
- Yoshiteru Oshima
- Haruhisa Kikuchi
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Affiliations: Research and Development Center, FUSO Pharmaceutical Industries, Ltd., Osaka 536‑8523, Japan, Department of Health Science and Social Welfare, Kibi International University, Takahashi, Okayama 716‑8508, Japan, Laboratory of Natural Products Chemistry, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Miyagi 980‑8578, Japan
Published online on: February 26, 2020 https://doi.org/10.3892/etm.2020.8553
Pages: 3150-3158

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Abstract

The therapeutic blockade of immune checkpoint has emerged as an effective treatment option for a broad range of tumors. However, the objective tumor response is still limited to a small number of cases and tumor types. The full utility of monoclonal antibody (mAb)‑based treatment is hindered by several inherent limitations. Thus, there is an urgent requirement to explore alternative modalities targeting the same pathways. In the present study, two amide analogues of brefelamide, TPFS‑201 and TPFS‑202, were identified as small molecular immune checkpoint inhibitors, as they downregulated PD‑L1 expression in tumor cells. PD‑L1 was suppressed in cancer cells treated with TPFD compounds at both mRNA and protein levels, as detected by reverse transcription quantitative PCR and flow cytometric analysis, respectively. Reporter assays using a PD‑L1 promoter luciferase construct confirmed the transcriptional inhibition of PD‑L1 by TPFS compunds. TPFS compound‑mediated PD‑L1 downregulation in cancer cells consequently restored T cell activity, as identified by the reduction of apoptosis and an increase in interleukin‑2 promoter activity in Jurkat T cells, which were co‑cultured with TPFS compound‑treated A549 cells. TPFS compound‑mediated PD‑L1 inhibition was partially abolished by the disruption of the putative transcriptional co‑activator with PDZ (TAZ)/TEA domain (TEAD)‑binding motif in the PD‑L1 promoter. The inhibitory effect of TPFS compounds on PD‑L1 was markedly inhibited in mouse cell lines, which is consistent with previous research demonstrating that PD‑L1 regulation by TAZ is not conserved in mice due to distinct promoter sequences flanking the TAZ/TEAD‑binding motif. Together, the data of the current study indicated the potential utility of the brefelamide amide analogues as small molecule immune checkpoint inhibitors, thereby providing therapeutic alternatives, which could be used as monotherapy or in combination with mAbs‑based treatment.

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