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Most conventional inhibitors are target\u2010specific and ineffective against broad\u2010spectrum viral infections. Therefore, the current scenario demands developing inhibitors to attenuate the membrane properties instead of targeting the viral proteins. In the present work, we have designed a new WD\u2010containing peptide (dmTG\u201023) by introducing a WD repeat at the center of the peptide sequence using mTG\u201023 peptide as a template and a 14\u2010amino acid peptide with seven WD repeats (WD\u201014), and tested their efficacies against the polyethylene glycol (PEG)\u2010induced fusion assay. Our findings show that both dmTG\u201023 and WD\u201014 prevent PEG\u2010induced fusion of small unilamellar vesicles (SUVs) with varying cholesterol concentrations, with a higher effect for dmTG\u201023. Moreover, we have compared our results with our previous observations on TG\u201023 and mTG\u201023 to evaluate the importance of the number and position of WD repeats in the peptide sequence to impart the highest inhibitory activity. Our results demonstrate that three WD repeat\u2010containing dmTG\u201023 shows the highest efficacy against the PEG\u2010induced fusion of SUVs by enhancing the membrane order at the acyl chain region. This result is significantly important in designing peptide\u2010based broad\u2010spectrum fusion inhibitors.<\/jats:p>","DOI":"10.1002\/asia.202500474","type":"journal-article","created":{"date-parts":[[2025,5,22]],"date-time":"2025-05-22T07:38:26Z","timestamp":1747899506000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Importance of the Number and Position of Tryptophan\u2013Aspartate Repeats in Designing Peptide\u2010Based Membrane Fusion Inhibitors"],"prefix":"10.1002","volume":"20","author":[{"given":"Manbit Subhadarsi","family":"Panda","sequence":"first","affiliation":[{"name":"School of Chemistry Sambalpur University  Jyoti Vihar Burla Odisha 768 019 India"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7499-5707","authenticated-orcid":false,"given":"Hirak","family":"Chakraborty","sequence":"additional","affiliation":[{"name":"School of Chemistry Sambalpur 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