UNL, CONICET, FBCB (School of Biochemistry and Biological Sciences), CBL (Biotechnological Center of Litoral), Ciudad Universitaria, Ruta Nacional 168 - Km 472.4 - C.C. 242 - (S3000ZAA) Santa Fe, Argentina.
UNL, CONICET, FBCB (School of Biochemistry and Biological Sciences), CBL (Biotechnological Center of Litoral), Ciudad Universitaria, Ruta Nacional 168 - Km 472.4 - C.C. 242 - (S3000ZAA) Santa Fe, Argentina.
J Biotechnol. 2019 Sep 10;303:46-52. doi: 10.1016/j.jbiotec.2019.07.006. Epub 2019 Jul 20.
Different strategies have been developed and successfully applied to biotherapeutics in order to improve their in vivo efficacy. The genetic fusion to natural or synthetic glycosylated peptides constitutes a promising strategy since it conserves the protein sequence and results in the improvement of the pharmacokinetic properties. The ANITVNITV peptide described by Perlmann and coworkers presents 9 amino acids and 2 potential N-glycosylation sites. Its fusion to FSH resulted in the increase of the molecular mass and negative charge of the protein. Consequently, the pharmacokinetics was considerably improved. The aim of the present study was to compare the influence of ANITVNITV peptide fusion on the physicochemical, biological and pharmacokinetic properties of native hIFN-α2b (IFNwt), which contains a single O-glycosylation site, and a hyperglycosylated variant (IFN4N), that bears, in addition, 4 N-linked glycans. The resulting molecules, IFNwtNter and IFN4NNter, evidenced a higher molecular mass and negative charge compared to IFNwt and IFN4N, respectively. Therefore, the pharmacokinetic properties of the new molecules were significantly improved. The molecules obtained by the synthetic peptide fusion strategy evidenced a decrease in their in vitro antiviral specific biological activities (SBA). However, in vitro antiproliferative SBA was differentially modified for IFNwtNter and IFN4NNter in comparison with the parental molecules. For IFNwtNter, a reduction in the antiproliferative SBA was also observed. Remarkably, the addition of the ANITVNITV peptide to the N-terminus of IFN4N had a positive impact on its growth-inhibitory activity. This feature together with its improved pharmacokinetics encourages the development of IFN4NNter as an IFN-α based biobetter.
为了提高生物治疗药物的体内疗效，已经开发并成功应用了不同的策略。将天然或合成糖基化肽与蛋白质进行基因融合是一种很有前途的策略，因为它保留了蛋白质序列，并改善了药物代谢动力学特性。Perlmann 及其同事描述的 ANITVNITV 肽由 9 个氨基酸和 2 个潜在的 N-糖基化位点组成。它与 FSH 的融合导致蛋白质的分子量和电荷增加。因此，药代动力学得到了显著改善。本研究的目的是比较 ANITVNITV 肽融合对天然 hIFN-α2b（IFNwt）和高糖基化变体（IFN4N）理化性质、生物学和药代动力学性质的影响。IFNwt 和 IFN4N 分别含有 1 个 O-糖基化位点和 4 个 N-糖基化位点。与 IFNwt 和 IFN4N 相比，得到的 IFNwtNter 和 IFN4NNter 分子具有更高的分子量和电荷。因此，新分子的药代动力学性质得到了显著改善。与 IFNwt 和 IFN4N 相比，通过合成肽融合策略获得的分子其体外抗病毒特定生物学活性（SBA）降低。然而，与亲本分子相比，IFNwtNter 和 IFN4NNter 的体外增殖抑制 SBA 发生了不同的变化。对于 IFNwtNter，也观察到其增殖抑制 SBA 降低。值得注意的是，将 ANITVNITV 肽添加到 IFN4N 的 N 末端对其生长抑制活性有积极影响。这种特性及其改善的药代动力学特性鼓励开发 IFN4NNter 作为基于 IFN-α 的生物改良药物。
