en بیوانفورماتیک Bioinformatic پژوهشي Research <strong>Background</strong>: Diphtheria toxin (DT) and its non-toxic mutant, CRM197, possess anti-cancer properties. The membrane-anchored form of heparin-binding epidermal growth factor-like growth factor (HBEGF) is a cellular receptor for DT and CRM197. The R and T domains of DT and CRM197 are responsible for receptor binding and translocation of their catalytic domains to the cytosol, respectively. Moreover, HB-EGF is a ligand of the epidermal growth factor receptor (EGFR), which is involved in the signaling cascade for cancer progression. HB-EGF binds to EGFR at its EGF-like domain. This study aimed to design anti-cancer peptides based on interactions of DT and CRM197 with HB-EGF to inhibit the binding of HB-EGF to EGFR and thus prevent the signaling involved in cancer progression.<br> <strong>Materials and Methods:</strong> The DT-HB-EGF and the CRM197-HB-EGF complexes were analyzed using molecular dynamics (MD) simulations at 60 ns. The stability, allergenicity and antigenicity of peptides were assessed using Protparam, AlgPred, and Vaxijen softwares respectively.<br> <strong>Results</strong>: The root mean square fluctuation (RMSF) plots demonstrated that the amino acids of DT and CRM197 with lower fluctuations were located in the R and T domains. The Rg plots revealed that the structure of the DT-HB-EGF complex remained compact until 31 ns, after which its compactness decreased. The structure of CRM197-HB-EGF remained compact during simulations. The root mean square deviation (RMSD) plots indicated that the DT-HB-EGF complex remained stable for 31 ns. However, the CRM197-HB-EGF complex produced a stable trajectory during simulations. Assessing tertiary structures of the complexes showed that residues Ser508-Ser535 of DT (pDT) and Ser291-Tyr358 of CRM197 (pCRM) were bound to the EGF-like domain of HBEGF; therefore, these peptides can interfere with the binding of HB-EGF to EGFR. The pCRM peptide was located in the T domain of CRM197, while the pDT peptide was located within the R domain. The Protparam results showed that both peptides were stable. However, pCRM was more thermostable than pDT. None of the peptides was allergens, as revealed by AlgPred. The Vaxijen analysis demonstrated that pCRM was non-antigenic, whereas pDT was antigenic.<br> <strong>Conclusion</strong>: These results demonstrate the utility of pCRM as an anticancer peptide that inhibits HB-EGF binding to EGFR.<br> &nbsp; Anti-cancer peptides, Diphtheria toxin (DT), Epidermal growth factor receptor (EGFR), Heparin-binding epidermal growth factor-like growth factor (HB-EGF), Molecular dynamics (MD)simulations 131 140 http://rmm.mazums.ac.ir/browse.php?a_code=A-10-1441-2&slc_lang=en&sid=1 Soheila Ghaderi soheilaghaderi7@gmail.com 100319475328460013572 100319475328460013572 No Department of Central Laboratory, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran. Shirin Tarahomjoo starahomjoo@gmail.com 100319475328460013573 100319475328460013573 Yes Department of Antigen Studies, Razi Vaccine and Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Karaj, Iran.