The present study was designed to evaluate the nephroprotective effect of PHYMIN-22 in human embryonic kidney (HEK)-293 cells and in-silico experiments. Invitro studies were investigated in HEK-293 cells to assess the cytoproliferative potential of PHYMIN-22 with cisplatin (CP) as a nephrotoxic agent. Further, Molecular docking studies in silico experiment were conducted to find an understanding of the potential interplay between Hsp90 and RIP1kinase proteins and PHYMIN-22. Treatment with PHYMIN-22 alone did not cause any considerable effect on cell survival. However, the cisplatin (20 μM) treatment induces significant (P < 0.001) effect on the HEK-293 cell viability. PHYMI-22 treatment significantly (P < 0.001) increased the cell viability at concentration of 125 and 250 μg/ mL. The microscopical results depict that cisplatin treatment induce irregular morphology and cell death, PHYMIN-22 treatment revert the irregular morphology into normal. The DNA fragmentation is considered as a hallmark of cytotoxicity, Slight DNA smash/damage has observed in cisplatin (20 μM) treated cells, after the treatment of PHYMIN-22 by different concentration (62.5, 125 and 250 μg/ml) reduced the DNA fragmentation to normal. In molecular docking insilico experiment the docking score values indicates that, the PHYMIN22 have high docking in negative with HSP90 as -7.6 kcal/mol. Compared with RIP1 kinase protein, it has high binding affinity. Therefore, PHYMIN-22 treatment was confirmed to harbor proliferative and anti-apoptotic effects in Hek-293 cells with lower cytotoxicity by its cytoprotective functions.