Battah Sinan, O'Neill Sophie, Edwards Christine, Balaratnam Sherina, Dobbin Paul, MacRobert Alexander J
National Medical Laser Centre, Division of Surgical and Interventional Sciences, Royal Free and University College Medical School, University College London, Charles Bell House, 67-73 Riding House St, London W1W 7EJ, UK.
Int J Biochem Cell Biol. 2006;38(8):1382-92. doi: 10.1016/j.biocel.2006.02.001. Epub 2006 Mar 3.
Intracellular porphyrin generation following administration of 5-aminolaevulinic acid has been widely used in photodynamic therapy for a range of malignant and certain non-malignant lesions. However, cellular uptake of 5-aminolaevulinic acid is limited by its hydrophilic nature and improved means of delivery are therefore being sought. Highly branched polymeric drug carriers known as dendrimers are a promising new approach to drug delivery. The aim of this study was to investigate the efficacy of dendrimers conjugated with 5-aminolaevulinic acid for porphyrin production in the transformed PAM 212 keratinocyte cell line and skin explants. Each dendritic derivative incorporated three 5-aminolaevulinic acid residues which were conjugated as esters via methyl or propyl linkers to a central tertiary carbon whose remaining terminal bore an amino, aminobenzyloxycarbonyl or nitro group. In the cell line, all compounds were more efficient at low concentrations compared to equimolar 5-aminolaevulinic acid for porphyrin production, with the most efficient incorporating the longer propyl linker. This compound was also the most lipophilic according to partition coefficient measurements. The intracellular porphyrin fluorescence levels showed good correlation with cellular phototoxicity following light exposure for all the compounds, together with minimal dark toxicity. Our findings indicate that the key factors influencing the efficacy of the dendritic derivatives are lipophilicity and steric hindrance within the dendritic structure which could restrict access to intracellular esterases for liberation of 5-aminolaevulinic acid. These findings should be taken into account in the design of larger dendrimers of 5-aminolaevulinic acid.
5-氨基乙酰丙酸给药后细胞内卟啉生成已广泛应用于一系列恶性和某些非恶性病变的光动力治疗。然而,5-氨基乙酰丙酸的细胞摄取受到其亲水性的限制,因此正在寻求改进的给药方式。被称为树枝状聚合物的高度分支聚合物药物载体是一种很有前景的新型药物递送方法。本研究的目的是研究与5-氨基乙酰丙酸共轭的树枝状聚合物在转化的PAM 212角质形成细胞系和皮肤外植体中产生卟啉的功效。每种树枝状衍生物都包含三个5-氨基乙酰丙酸残基,它们通过甲基或丙基连接基作为酯与中心叔碳共轭,中心叔碳的其余末端带有氨基、氨基苄氧基羰基或硝基。在细胞系中,与等摩尔的5-氨基乙酰丙酸相比,所有化合物在低浓度下产生卟啉的效率更高,其中效率最高的含有较长的丙基连接基。根据分配系数测量,该化合物也是亲脂性最强的。对于所有化合物,细胞内卟啉荧光水平与光照后的细胞光毒性显示出良好的相关性,同时暗毒性最小。我们的研究结果表明,影响树枝状衍生物功效的关键因素是树枝状结构内的亲脂性和空间位阻,这可能会限制细胞内酯酶释放5-氨基乙酰丙酸的途径。在设计更大的5-氨基乙酰丙酸树枝状聚合物时应考虑这些研究结果。