Efficient Orthogonal Bioconjugation of Dendrimers for Synthesis of Bioactive Nanoparticles
Efficient Orthogonal Bioconjugation of Dendrimers for Synthesis of Bioactive Nanoparticles
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DOI：
10.1021/bc1005653
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发表时间：
2011-06-01
影响因子：
4.7
通讯作者：
Hartley, Oliver
中科院分区：
文献类型：
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作者：
Gaertner, Hubert F.;Cerini, Fabrice;Hartley, Oliver
携带生物活性功能组（例如，靶向部分、有效载荷、示踪剂）在广泛的临床应用中具有潜在的用途。虽然复杂，但这种结构应尽可能具有确定的分子结构并且是单分散的。然而，实现该目标的现有方法不适合于肽和蛋白质的掺入，并且提供两种不同类型的功能元件的正交引入的那些方法与使用市售材料不相容。在这项研究中，我们已经开发了基于市售聚酰胺胺（PAMAM）树枝状聚合物的纳米颗粒的生产方法。首先，我们确定了一种优化的肟缀合策略，在该策略下，复杂的树枝状聚合物不仅可以用模型肽，而且可以用重组蛋白质（胰岛素作为例子）完全修饰。其次，我们开发了一种基于胱胺核心PAMAM树状聚合物的双链共价杂树枝状聚合物（“二嵌段”）的策略，并将其用于生成杂树枝状聚合物，其中肽阵列和甘露糖阵列被正交引入。最后，通过将官能化连接体并入二嵌段结构中，我们能够将第三功能元件位点特异性地引入纳米颗粒中。我们使用荧光素、甘露糖阵列和肽阵列作为三种功能来举例说明这种方法。我们表明，将甘露糖阵列掺入纳米颗粒中强烈且特异性地增强了免疫系统的哨兵细胞的摄取，这是疫苗递送应用的重要特性。这些基于PAMAM树枝状聚合物的方法代表了生物活性纳米颗粒开发的强大和通用的平台。
Nanoparticles carrying biologically active functional sets (e.g., targeting moiety, payload, tracer) have potential use in a wide range of clinical applications. Though complex, such constructions should, as far as possible, have a defined molecular architecture and be monodisperse. However, the existing methods to achieve this goal are unsuitable for the incorporation of peptides and proteins, and those that provide for orthogonal introduction of two different types of functional element are incompatible with the use of commercially available materials. In this study, we have developed approaches for the production of nanoparticles based on commercially available polyamidoamine (PAMAM) dendrimers. First, we identified an optimized oxime conjugation strategy under which complex dendrimers can be fully decorated not only with model peptides, but also with recombinant proteins (insulin was taken as an example). Second, we developed a strategy based on a two-chain covalent heterodendrimer (a "diblock") based on cystamine core PAMAM dendrimers and used it to generate heterodendrimers, into which a peptide array and a mannose array were orthogonally introduced. Finally, by incorporating a functionalized linker into the diblock architecture we were able to site-specifically introduce a third functional element into the nanoparticle. We exemplified this approach using fluorescein, a mannose array, and a peptide array as the three functionalities. We showed that incorporation of a mannose array into a nanoparticle strongly and specifically enhances uptake by sentinel cells of the immune system, an important property for vaccine delivery applications. These PAMAM dendrimer-based approaches represent a robust and versatile platform for the development of bioactive nanoparticles.