Peramivir, an Anti-Influenza Virus Drug, Exhibits Potential Anti-Cytokine Storm Effects

Abstract

Coronavirus Disease 2019 (COVID-19) infected by Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2) has been declared a public health emergency of international concerns. Cytokine storm syndrome (CSS) is a critical clinical symptom of severe COVID-19 patients, and the macrophage is recognized as the direct host cell of SARS-CoV-2 and potential drivers of CSS. In the present study, peramivir was identified to reduce TNF-α by partly intervention of NF-κB activity in LPS-induced macrophage model. In vivo, peramivir reduced the multi-cytokines in serum and bronchoalveolar lavage fluid (BALF), alleviated the acute lung injury and prolonged the survival time in mice. In human peripheral blood mononuclear cells (hPBMCs), peramivir could also inhibit the release of TNF-α. Collectively, we proposed that peramivir might be a candidate for the treatment of COVID-19 and other infections related CSS.

Keywords: COVID-19; acute lung injury; cytokine storm syndrome; multi-cytokines; peramivir.

Graphical Abstract

Figure 1

Identification of peramivir as anti-inflammatory agents. (A) Chemical structures of peramivir, oseltamivir and zanamivir. (B) Peramivir showed the strongest TNF-α inhibitory effect compared with oseltamivir and zanamivir, **P < 0.01, ***P < 0.001 vs peramivir. (C) The dose-response curves for the TNF-α inhibitions of peramivir exhibited IC₅₀s of 4.3 µM. (D) Cell viabilities of macrophages with peramivir treatment at different concentrations. N = 3.

Figure 2

Peramivir has a significant effect on some cytokines in mouse serum and bronchoalveolar lavage fluid (BALF). (A–H) Serum cytokines. (I, J) BALF cytokines. *P < 0.05, **P <0.01, ***P < 0.001, ****P < 0.0001. N = 8-10.

Figure 3

The weak effect of peramivir on chemokines and IL -10 in the serum of mice. (A–C) serum chemokines: CXCL1, CCL5, CXCL10. (D) serum cytokines: IL-10. ns, no significance. N = 10.

Figure 4

The weak effect of peramivir on some cytokines in the bronchoalveolar lavage fluid (BALF) of mice. (A, B) BALF cytokines: IFN-γ, IFN-β. (C) BALF chemokines: MCP-1. (D) BALF cytokines: GM-CSF. (E–J) BALF cytokines: IL-1α, IL-1β, IL-10, IL-17A, IL-27, IL-23. ns, no significance. N = 8-10.

Figure 5

Peramivir effectively attenuates acute lung injury and prolong the survival in LPS-induced mice. (A) Representative images of lung H&E staining of control, and peramivir treatment groups. Black, green and yellow arrows indicated infiltration of inflammatory cells, congestion and edema within thickened alveolar, respectively. Scale bars, 100 or 200 μm as indicated. (B) Lung injury scores of control and peramivir treatment groups (n=5). *P < 0.05. (C) Survival time of LPS-induced CSS in control, and peramivir (20, 60 mg/kg) groups (n=10). Kaplan–Meier analysis was performed. *P < 0.05, **P < 0.01. (D) RAW264.7 cells were co-cultured with peramivir at concentrations of 2.5, 5 and 10 μM at 1 h before LPS stimulation. The activity of NF-κB luciferase was upregulated in all groups after 8 h, and there was a significant decline in cells co-cultured with peramivir in a dose-dependent manner. ***P < 0.001. (E) The activation of the NF-κB, MAPK and STAT pathway in LPS-stimulated macrophages after the treatment of peramivir. (F) p65 nuclear translocation in LPS-stimulated macrophages after the treatment of peramivir (blue, DAPI; green, p65; cyan, cyan). Scale bars, 10 μm as indicated.

Figure 6

Peramivir inhibits cytokine release in LPS-induced hPBMCs from the two health donors. TNF-α concentration was elevated by LPS stimulation. Peramivir reduced TNF-α release at the time 6h (A, C) and 12h (B, D) with dose (2.5, 5, 10 μM)-dependent manner. Peramivir showed no toxicity toward hPBMCs. *P < 0.05, **P < 0.01, *** P < 0.001.