Biessen Erik A L, Sliedregt-Bol Karen, 'T Hoen Peter A Chr, Prince Perry, Van der Bilt Erica, Valentijn A Rob P M, Meeuwenoord Nico J, Princen Hans, Bijsterbosch Martin K, Van der Marel Gijs A, Van Boom Jacques H, Van Berkel Theo J C
Division of Biopharmaceutics, Leiden/Amsterdam Center for Drug Research, Leiden University, P.O. Box 9503, 2300 RA Leiden, The Netherlands.
Bioconjug Chem. 2002 Mar-Apr;13(2):295-302. doi: 10.1021/bc015550g.
In this study, we present the design and synthesis of an antisense peptide nucleic acid (asPNA) prodrug, which displays an improved biodistribution profile and an equally improved capacity to reduce the levels of target mRNA. The prodrug, K(GalNAc)(2)-asPNA, comprised of a 14-mer sequence complementary to the human microsomal triglyceride transfer protein (huMTP) gene, conjugated to a high-affinity tag for the hepatic asialoglycoprotein receptor (K(GalNAc)(2)). The prodrug was avidly bound and rapidly internalized by HepG2s. After iv injection into mice, K(GalNAc)(2)-asPNA accumulated in the parenchymal liver cells to a much greater extent than nonconjugated PNA (46% +/- 1% vs 3.1% +/- 0.5% of the injected dose, respectively). The prodrug was able to reduce MTP mRNA levels in HepG2 cells by 35-40% (P < 0.02) at 100 nM in an asialoglycoprotein receptor- and sequence-dependent fashion. In conclusion, hepatocyte-targeted PNA prodrugs combine a greatly improved tropism with an enhanced local intracellular availability and activity, making them attractive therapeutics to lower the expression level of hepatic target genes such as MTP.
在本研究中，我们展示了一种反义肽核酸（asPNA）前药的设计与合成，其具有改善的生物分布特征以及同等程度改善的降低靶标mRNA水平的能力。该前药K(GalNAc)(2)-asPNA由与人类微粒体甘油三酯转运蛋白（huMTP）基因互补的14聚体序列组成，并与肝去唾液酸糖蛋白受体的高亲和力标签（K(GalNAc)(2)）偶联。该前药被HepG2细胞强烈结合并迅速内化。静脉注射到小鼠体内后，K(GalNAc)(2)-asPNA在实质肝细胞中的蓄积程度比未偶联的PNA高得多（分别为注射剂量的46%±1%和3.1%±0.5%）。该前药能够以去唾液酸糖蛋白受体和序列依赖性方式在100 nM时将HepG2细胞中的MTP mRNA水平降低35 - 40%（P < 0.02）。总之，肝细胞靶向的PNA前药将大大改善的靶向性与增强的局部细胞内可用性和活性相结合，使其成为降低诸如MTP等肝靶标基因表达水平的有吸引力的治疗药物。
