Synthesis of a new peptide and evaluation of its cytotoxicity and regenerative properties by prediction and in vitro experiment

“Oral bioavailability is an important molecular descriptor in the development of BAS for absorption and metabolism during the first passage through the liver [24].”

“Cell viability was assessed using the 3-(4,5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide (MTT) assay [17].”

“The MMP protein plays an important role in maintaining and restoring tissue homeostasis after tissue damage and performs a wide range of functions [36].”

“Inactively expressed in a latent complex with a latent-associated peptide (LAP), TGFß1 is secreted into the extracellular space and bound by latent TGFß-binding protein 1 (LTBP1), where it remains available for activation [41].”

“For the studied peptide, it is 0.869 with an optimal value for medicinal substances ranging from 0.5 to 3 [28].”

“This growth factor has been identified as a regulator not only of myofibroblast resorption but also of myofibroblast formation during wound healing, among other processes [39,40].”

“Thus, the cyclic peptide CARSKNKDC with an active sequence of SK and/or KS with cyclization through C1-C2 bonds restores damaged blood vessels during angiogenesis in inflammatory diseases, whose receptors are heparan sulfate proteoglycans [12].”

“In addition, the authors [37] analyzed the transcriptomics of migration genes in inflammation, in which the predominant localization of MMP3 was established—in the direction of the wound bed versus the wound edge or intact skin.”

“Hussain et al. [13] came to similar conclusions when they investigated the cyclic peptide CARGGLKSC.”

“The data obtained are consistent with studies [32] in which a decrease in the inflammatory process in the lungs, joints, and intestine was found in animal models on the background of migration of COL3A1 proteins.”

“This peptide consists of two threonine (T) molecules [19].”

“Despite their effectiveness, peptides remain difficult to create (predict and design) because bioactivity is influenced by many characteristics, including structure, number of amino acids, sequence, molecular weight, total charge, hydrophobicity, lipophilicity, and so on [7].”

“At the same time, many new peptides have been developed, but only a few of them have regenerative properties [8].”

“A study using monolayers of Caco-2 cells showed that optimal permeability of the compound is achieved at logP values of 4 to 5, while increasing the logP value outside this range decreases the permeability of the preparations [26].”

“The development of a peptide with regenerative properties was based on an analysis of the scientific literature [12–14].”

“One of the mechanisms of action of regenerative peptides, as evidenced by data [20,21], maybe as follows: the peptide regulates inflammation by physically interacting with collagen fibers and serving as a structural component of collagen fibers, thereby affecting cellular functions such as…”

“According to the authors [1–3], some peptide sequences have pronounced regenerative properties.”

“The indicator logP ≤0 indicates high hydrophilicity and logP >0 indicates increased lipophilicity [25].”

“Some peptides target bone tissue or fractures, binding to either tartrate-resistant acid phosphatase deposited by osteoclasts or hydroxyapatite deposited by osteoblasts [6].”

“The peptide was identified by MALDI-TOF MS Ultraflex method (Bruker, Germany) [16].”

“The authors [4] claim that peptides can bind to collagen in the extracellular matrix (ECM), which is exposed during inflammation.”

“Fibroblast migration can accelerate the process of wound reepithelialization and promote wound closure during the healing process [30].”

“Our results suggest that COL3A1 can be used as a diagnostic biomarkers of inflammation in tissue damage, which is consistent with studies [35], in which an increase in COL3A1 expression was found in joint injuries.”

“COL3A1 genes were identified in inflammatory processes and their regenerative function was confirmed using Funrich software [31].”

“…indicator reflects the distribution of n -octanol in a buffer solution at pH = 7.4 and characterizes the lipophilicity/hydrophobicity balance of the test substance and influences the effective penetration through the cell membrane, absorption, distribution, metabolism, excretion, and toxicity [27].”

“The obtained predicted IC 50 values are important for the assessment of cardiac toxicity of new biologically active compounds [18].”

“In the studies of the authors [33,34], it was proved that COL3A1 expression levels increased at the early stages of inflammatory processes, in particular with damage to compounds, and decreased at later stages.”

“This growth factor has been identified as a regulator not only of myofibroblast resorption but also of myofibroblast formation during wound healing, among other processes [39,40].”

“The data obtained on the physicochemical characteristics of the CC-18 peptide demonstrate that it is bioavailable, which is consistent with the empirical findings of other researchers [22,23].”

“In studies [38] on laboratory mice with excised wounds, the compensatory role of MMP3 expression was demonstrated with a decrease in MMP14 in adult skin fibroblasts (MMP14Sf-/-), leading to accumulation of type I collagen and increased MMP3 expression.”

“Therefore, it is difficult to design peptides that can selectively bind to the desired target while avoiding non-target interactions [10].”

“In the studies of the authors [33,34], it was proved that COL3A1 expression levels increased at the early stages of inflammatory processes, in particular with damage to compounds, and decreased at later stages.”

“The data obtained on the physicochemical characteristics of the CC-18 peptide demonstrate that it is bioavailable, which is consistent with the empirical findings of other researchers [22,23].”

“The molecular weight distribution of the peptide was evaluated by mass spectrometry [15].”

“The incorporation of a peptide fragment into a known stable cyclic peptide framework [11].”

“The ML-based tool for ranking peptides, named PeptideRanker, can also assist with high-throughput screening, ranking peptides according to their likelihood to be biologyically active so that researchers prioritize sequences to be synthesized chemically or tested in vivo [210].”

“Bioinformatics tools, such as PeptideRanker and the Antimicrobial Peptide Calculator and Predictor enable the prediction of bioactive peptides and their potential antimicrobial activity based on amino acid sequences (Tikhonov et al., 2025).”