Abstract <p>Ergosterol peroxide (EP) is a natural product with a wide range of biological activities. To obtain compounds with improved antitumor activity, a series of 15 EP-3-piperazine carbamate indazole derivatives were designed and synthesized using carbamate as a linker. The new compounds were evaluated for their cytotoxic activities against the human lung cancer A549 cell line, human liver cancer HepG2 cell line, and human breast cancer MCF-7 cell line. Among these derivatives, (3β,5α,8α,22<i>E</i>)-5,8-epidioxyergosta-6,22-dien-3-yl-4-[(1<i>H</i>-indazol-7-yl)methyl]piperazine-1-carboxylate showed the strongest antiproliferative activity, with an IC₅₀ value of 2.74 μM against A549 cells, which was superior to those of EP (IC₅₀ = 19.42 μM) and cisplatin (IC₅₀ = 5.82 μM). The molecular docking results indicated that the (3β,5α,8α,22<i>E</i>)-5,8-epidioxyergosta-6,22-dien-3-yl 4-[(1<i>H</i>-indazol-7-yl)methyl]piperazine-1-carboxylate –VEGFR protein binding energy was 10.49 kcal/mol, and further validation at the molecular level through Western blot analysis demonstrated that it could significantly inhibit the expression of the P-VEGFR protein. (3β,5α,8α,22<i>E</i>)-5,8-Epidioxyergosta-6,22-dien-3-yl-4-[(1<i>H</i>-indazol-7-yl)methyl]piperazine-1-carboxylate is a potential anti-VEGFR protein inhibitor, the antitumor activity of which we aim to further evaluate in vivo (i.e. in mice).</p>