*Article* **Anti-Cancer Effects of Cyclic Peptide ALOS4 in a Human Melanoma Mouse Model**

**Bar Levi 1,†, Shiri Yacobovich 1,†, Michael Kirby <sup>1</sup> , Maria Becker <sup>2</sup> , Oryan Agranyoni <sup>1</sup> , Boris Redko <sup>3</sup> , Gary Gellerman <sup>3</sup> , Albert Pinhasov 1,2, Igor Koman <sup>4</sup> and Elimelech Nesher 1,4,\***


**Abstract:** We examined the effects of ALOS4, a cyclic peptide discovered previously by phage library selection against integrin αvβ<sup>3</sup> , on a human melanoma (A375) xenograft model to determine its abilities as a potential anti-cancer agent. We found that ALOS4 promoted healthy weight gain in A375-engrafted nude mice and reduced melanoma tumor mass and volume. Despite these positive changes, examination of the tumor tissue did not indicate any significant effects on proliferation, mitotic index, tissue vascularization, or reduction of αSMA or Ki-67 tumor markers. Modulation in overall expression of critical downstream αvβ<sup>3</sup> integrin factors, such as FAK and Src, as well as reductions in gene expression of *c-Fos* and *c-Jun* transcription factors, indirectly confirmed our suspicions that ALOS4 is likely acting through an integrin-mediated pathway. Further, we found no overt formulation issues with ALOS4 regarding interaction with standard inert laboratory materials (polypropylene, borosilicate glass) or with pH and temperature stability under prolonged storage. Collectively, ALOS4 appears to be safe, chemically stable, and produces anti-cancer effects in a human xenograft model of melanoma. We believe these results suggest a role for ALOS4 in an integrin-mediated pathway in exerting its anti-cancer effects possibly through immune response modulation.

**Keywords:** cancer; cyclic peptide; integrin; αvβ<sup>3</sup> ; ALOS4; melanoma
