Division of Pharmaceutical Sciences, School of Pharmacy, University of Missouri-Kansas City, 2464 Charlotte Street, Kansas City, MO 64108-2718, USA.
Int J Pharm. 2012 Sep 15;434(1-2):315-24. doi: 10.1016/j.ijpharm.2012.05.033. Epub 2012 Jun 9.
A majority of studies involving prodrugs are directed to overcome low bioavailability of the parent drug. The aim of this study is to increase the bioavailability of acyclovir (ACV) by designing a novel prodrug delivery system which is more lipophilic, and at the same time site specific. In this study, a lipid raft has been conjugated to the parent drug molecule to impart lipophilicity. Simultaneously a targeting moiety that can be recognized by a specific transporter/receptor in the cell membrane has also been tethered to the other terminal of lipid raft. Targeted lipid prodrugs i.e., biotin-ricinoleicacid-acyclovir (B-R-ACV) and biotin-12hydroxystearicacid-acyclovir (B-12HS-ACV) were synthesized with ricinoleicacid and 12hydroxystearicacid as the lipophilic rafts and biotin as the targeting moiety. Biotin-ACV (B-ACV), ricinoleicacid-ACV (R-ACV) and 12hydroxystearicacid-ACV (12HS-ACV) were also synthesized to delineate the individual effects of the targeting and the lipid moieties. Cellular accumulation studies were performed in confluent MDCK-MDR1 and Caco-2 cells. The targeted lipid prodrugs B-R-ACV and B-12HS-ACV exhibited much higher cellular accumulation than B-ACV, R-ACV and 12HS-ACV in both cell lines. This result indicates that both the targeting and the lipid moiety act synergistically toward cellular uptake. The biotin conjugated prodrugs caused a decrease in the uptake of [(3)H] biotin suggesting the role of sodium dependent multivitamin transporter (SMVT) in uptake. The affinity of these targeted lipid prodrugs toward SMVT was studied in MDCK-MDR1 cells. Both the targeted lipid prodrugs B-R-ACV (20.25 ± 1.74 μM) and B-12HS-ACV (23.99 ± 3.20 μM) demonstrated higher affinity towards SMVT than B-ACV (30.90 ± 4.19 μM). Further, dose dependent studies revealed a concentration dependent inhibitory effect on [(3)H] biotin uptake in the presence of biotinylated prodrugs. Transepithelial transport studies showed lowering of [(3)H] biotin permeability in the presence of biotin and biotinylated prodrugs, further indicating a carrier mediated translocation by SMVT. Overall, results from these studies clearly suggest that these biotinylated lipid prodrugs of ACV possess enhanced affinity towards SMVT. These prodrugs appear to be potential candidates for the treatment of oral and ocular herpes virus infections, because of higher expression of SMVT on intestinal and corneal epithelial cells. In conclusion we hypothesize that our novel prodrug design strategy may help in higher absorption of hydrophilic parent drug. Moreover, this novel prodrug design can result in higher cell permeability of hydrophilic therapeutics such as genes, siRNA, antisense RNA, DNA, oligonucleotides, peptides and proteins.
大多数涉及前药的研究旨在克服母体药物生物利用度低的问题。本研究旨在通过设计一种新的前药传递系统来提高阿昔洛韦(ACV)的生物利用度,该系统具有更高的亲脂性,同时具有特定的部位特异性。在本研究中,脂质筏已与母体药物分子缀合以赋予亲脂性。同时,还将可以被细胞膜上特定转运体/受体识别的靶向部分连接到脂质筏的另一端。合成了靶向脂质前药,即生物素-蓖麻酸-阿昔洛韦(B-R-ACV)和生物素-12-羟基硬脂酸-阿昔洛韦(B-12HS-ACV),其中蓖麻酸和 12-羟基硬脂酸作为亲脂性筏,生物素作为靶向部分。还合成了生物素-ACV(B-ACV)、蓖麻酸-ACV(R-ACV)和 12-羟基硬脂酸-ACV(12HS-ACV),以阐明靶向和脂质部分的单独作用。在汇合的 MDCK-MDR1 和 Caco-2 细胞中进行了细胞积累研究。靶向脂质前药 B-R-ACV 和 B-12HS-ACV 在两种细胞系中的细胞积累均明显高于 B-ACV、R-ACV 和 12HS-ACV。这一结果表明,靶向和脂质部分都协同作用于细胞摄取。生物素缀合的前药会导致 [(3)H]生物素摄取减少,表明钠离子依赖的多种维生素转运体(SMVT)在摄取中起作用。在 MDCK-MDR1 细胞中研究了这些靶向脂质前药对 SMVT 的亲和力。靶向脂质前药 B-R-ACV(20.25 ± 1.74 μM)和 B-12HS-ACV(23.99 ± 3.20 μM)对 SMVT 的亲和力均高于 B-ACV(30.90 ± 4.19 μM)。此外,剂量依赖性研究显示,在存在生物素化前药的情况下,对 [(3)H]生物素摄取有浓度依赖性抑制作用。跨上皮转运研究表明,在存在生物素和生物素化前药的情况下,[(3)H]生物素的通透性降低,进一步表明 SMVT 通过载体介导的转位。总的来说,这些研究结果清楚地表明,ACV 的这些生物素化脂质前药对 SMVT 具有更高的亲和力。由于 SMVT 在肠上皮细胞和角膜上皮细胞中的高表达,这些前药可能成为治疗口腔和眼部疱疹病毒感染的潜在候选药物。综上所述,我们假设我们的新型前药设计策略可能有助于提高亲水性母体药物的吸收。此外,这种新型前药设计可以提高亲水治疗剂(如基因、siRNA、反义 RNA、DNA、寡核苷酸、肽和蛋白质)的细胞通透性。