A Potent d-Protein Antagonist of VEGF-A is Nonimmunogenic, Metabolically Stable, and Longer-Circulating in Vivo

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• 作者：Maruti Uppalapati ; Dong Jun Lee ; Kalyaneswar Mandal ; Hongyan Li ; Les P. Miranda ; Joshua Lowitz ; John Kenney ; Jarrett J. Adams ; Dana Ault-Riché ; Stephen B. H. Kent ; Sachdev S. Sidhu
• 刊名：ACS Chemical Biology
• 出版年：2016
• 出版时间：April 15, 2016
• 年：2016
• 卷：11
• 期：4
• 页码：1058-1065
• 全文大小：411K
• 年卷期：0
• ISSN：1554-8937

文摘

Polypeptides composed entirely of pan class="smallcaps">dpan>-amino acids and the achiral amino acid glycine (pan class="smallcaps">dpan>-proteins) inherently have in vivo properties that are proposed to be near-optimal for a large molecule therapeutic agent. Specifically, pan class="smallcaps">dpan>-proteins are resistant to degradation by proteases and are anticipated to be nonimmunogenic. Furthermore, pan class="smallcaps">dpan>-proteins are manufactured chemically and can be engineered to have other desirable properties, such as improved stability, affinity, and pharmacokinetics. Thus, a well-designed pan class="smallcaps">dpan>-protein therapeutic would likely have significant advantages over pan class="smallcaps">lpan>-protein drugs. Toward the goal of developing pan class="smallcaps">dpan>-protein therapeutics, we previously generated RFX001.D, a pan class="smallcaps">dpan>-protein antagonist of natural vascular endothelial growth factor A (VEGF-A) that inhibited binding to its receptor. However, RFX001.D is unstable at physiological temperatures (T_m = 33 °C). Here, we describe RFX037.D, a variant of RFX001.D with extreme thermal stability (T_m > 95 °C), high affinity for VEGF-A (K_d = 6 nM), and improved receptor blocking. Comparison of the two enantiomeric forms of RFX037 revealed that the pan class="smallcaps">dpan>-protein is more stable in mouse, monkey, and human plasma and has a longer half-life in vivo in mice. Significantly, RFX037.D was nonimmunogenic in mice, whereas the pan class="smallcaps">lpan>-enantiomer generated a strong immune response. These results confirm the potential utility of synthetic pan class="smallcaps">dpan>-proteins as alternatives to therapeutic antibodies.