The exploration of non-invasive drug administration routes, such as oral, nasal, ocular, and transdermal, has gained prominence as alternatives to traditional needle injections. Transdermal drug delivery stands out as an attractive method due to its ease of administration, sustained release, and minimized systemic side effects. In this study, we employ support vector machine-based models to design highly effective cell- penetrating peptides (CPPs), known for their efficient transduction and low cytotoxicity. A genomic construct, incorporating growth factors or segments of botulinum toxin, is synthesized and combined with a novel CPP named dermal penetration peptide (DPP). The induced expression of proteins is achieved through Isopropyl β-D-1-thiogalactopyranoside addition, followed by purification using Ni- NTA agarose resin and analysis through western blotting. Topical application of the newly designed peptides on human skin demonstrates their penetration ability, as confirmed through confocal imaging of frozen vertical skin sections. Skin penetration assays reveal a 3.3-fold increase in efficiency for DPP compared to the well-known CPP, TAT. The DPP-attached botulinum toxin exhibits equivalent cleavage efficacy to the non-linked counterpart in cell assays. Additionally, growth factor wound healing tests show significant wound recovery 48 hours post-treatment. Western blotting analysis confirms the expression of recombinant proteins, with specific bands detected at approximately 57.5 and 10 kDa. Cytotoxicity studies at high concentrations demonstrate the non-toxic nature of the developed materials. The results underscore the potential of DPP as a skin efficacy platform technology capable of producing substantial effects even in small quantities.
