Herein, we designed a nano peptide that contains three important motifs for targeting the chemotrypsin-like cysteine protease (3CLpro) which is the enzyme responsible for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) replication. The novel nano peptide contains the Nap Phe-Phe motif that is responsible for peptide self-assembly, an octapeptide (Ser-Ala-Val-Leu-Gln-Ser-Gly-Phe) motif where the enzyme recognizes the substrate and induces enzyme sensitivity, and a tetrapeptide motif which is positively charged containing the peptide (Lys)₄ that facilitates penetration into a cell. The nano peptide was characterized using Proton Nuclear Magnetic Resonance (H-NMR) and Liquid Chromatography-Mass Spectrometry (LC-MS) to confirm its structure. In vitro results showed that the presently formulated nano peptide was not cytotoxic to fibroblasts for up to 72 hours, bound to 3CLpro, inhibited SARS-CoV-2 Omicron variant virus replication, and was stable for binding for up to one week in culture. In this manner, this timely study demonstrates that this novel nano peptide should be studied for a wide range of Coronavirus Disease (COVID-19) prophylactic or therapeutic applications.