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Collagen hydrolysate is an excellent source of bioactive peptides and these gelatin-derived molecules can play a vital role in the treatment of tumors. Here, our objective was to investigate the effects of bovine collagen peptides (BCPs) on the proliferation and metastasis of ovarian cancer cells and identify peptides from bovine bone gelatin hydrolysate with anticancer activity against the human ovary carcinoma cell line, SKOV-3. The hydrolysate was initially purified using consecutive bioactivity-guided chromatography methods, with the resulting most active fraction subjected to LC-MS-MS analysis. Finally, 11 peptides were selected for functional validation. GPAGPQGPRGDKGETGEQGDR (all deamidated) were validated as having strong cytotoxicity against SKOV-3 cells and SKOV-3 cells bearing mice. Further, this peptide exhibited lower toxicity to MRC-5 cells and could inhibit cell invasion, and induce apoptosis. In addition, the underlying mechanism for the anticancer effect might be direct binding with α5β1 integrin suppressing JNK pathway.