Smart Biodegradable Nanoparticulate Materials: Poly-lactide-co-glycolide Functionalization with Selected Peptides
Smart Biodegradable Nanoparticulate Materials: Poly-lactide-co-glycolide Functionalization with Selected Peptides
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DOI：
10.2174/1573413712666151116213330
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发表时间：
2016-01-01
影响因子：
1.5
通讯作者：
Genta, Ida
中科院分区：
文献类型：
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作者：
Colzani, Barbara;Biagiotti, Marco;Genta, Ida
背景：智能纳米颗粒材料，即具有特定表面功能的定制纳米颗粒（NP），最近作为时间和位点特异性药物输送的有用工具而引起了人们的关注。具体来说，聚合物纳米颗粒（NP）可以用不同的化学实体（即肽）进行化学功能化，选择性地识别体内生物底物并靶向药物释放。为了获得肽修饰的纳米颗粒，可以采用不同且非常复杂的策略。方法：提出了一种简单的方法，用于用小肽直接功能化聚丙交酯-共-乙交酯（PLGA）。使用固相肽合成获得十二肽（GE11）和较小的四肽（FQPV）。 FQPV-和GE11-PLGA缀合物是通过优化碳二亚胺化学获得的。纳米沉淀和溶剂萃取/蒸发方法是专门为制备 FQPV-PLGA 纳米颗粒而设计的。纳米颗粒的特征包括尺寸、表面电荷和吸附的肽量；使用成体成纤维细胞对 FQPV-PLA NP 进行离体细胞毒性研究。结果：定制 GE11（最近被称为高效表皮生长因子受体靶向剂）和 FQPV（用作模型肽）通过固相肽合成合成，实现了良好的纯度（95％）和令人满意的工艺产率（70-85％）。然后，通过优化碳二亚胺化学获得 FQPV- 和 GE11-PLGA 缀合物，实现高度功能化 (> 85%)。意识到肽-PLGA 缀合物相对于普通 PLGA 的不同物理化学性质，专门构建了两种不同的 NP 制备技术，即纳米沉淀和溶剂萃取/蒸发方法，以制备 FQPV-PLGA NP。两种方法均表明适合获得具有适合肠胃外给药尺寸（<250nm）、窄尺寸分布（P.I.约0.1）、良好形态特征和负电荷（约-20mV）的纳米粒子。纳米颗粒上的肽吸附方案被认为是增加纳米颗粒表面肽表达的额外策略，旨在提高靶向效果。实验设计方法 (DoE) 已成功应用于更通用的溶剂萃取/蒸发方法，以系统地突出工艺参数（有机溶剂混合物、PVA 浓度和聚合物溶液体积）对 NP 尺寸的影响。结论：PLGA 成功地用两种不同的肽 FQPV 和 GE11 进行功能化，遵循通用且简单的碳二亚胺化学，无需进一步修改聚合物和/或肽结构。纳米沉淀和溶剂萃取/蒸发纳米颗粒制备方法都经过适当优化，以获得基于肽-PLGA的纳米颗粒，并且可以根据肽-聚合物缀合物和用于包封的药物的溶解度特性选择性地选择它们。预先形成的 PLGA NP 上的肽吸附可有效地用于增加 NP 表面上的肽表达，从而提高主动靶向情况下的细胞识别。对选定的基于肽-PLGA 的纳米颗粒材料的初步细胞相容性评估显示了用于制药目的的合成程序和纳米颗粒制备方法的潜在可行性。
Background: Smart nanoparticulate materials, namely tailored nanoparticles (NPs) with specific surface functionality, have recently attracted attention as useful tool for time-and site-specific drug delivery. Specifically, polymeric nanoparticles (NPs) can be chemically functionalized with different chemical entities, i.e. peptides, that selectively recognize biological substrates in vivo and target drug release. Divergent and very complex strategies can be pursued in order to obtain peptide-decorated NPs.Methods: A simple method was suggested for direct functionalization of poly-lactide-co-glicolide (PLGA) with small peptides. A solid-phase peptide synthesis was used to obtain a dodecapeptide (GE11) and a smaller tetrapeptide (FQPV). FQPV- and GE11-PLGA conjugates were obtained by optimized carbodiimide chemistry. Nanoprecipitation and solvent extraction/evaporation methods were purpose-built in order to prepare FQPV-PLGA NPs. NPs were characterized in terms of size, surface charge and adsorbed peptide amount; ex-vivo cytotoxicity studies were performed on FQPV-PLA NPs using adult fibroblasts.Results: Custom GE11, recently known as efficient Epidermal Growth Factor Receptor targeting agent, and FQPV, used as model peptide, were synthesized by solid-phase peptide synthesis achieving good purity (95%) and satisfactory process yields (70-85%). Then, FQPV- and GE11-PLGA conjugates were obtained by optimized carbodiimide chemistry achieving an high degree of functionalization (> 85%). Aware of different physico-chemical properties of peptide-PLGA conjugates with respect to plain PLGA, two different NPs preparation techniques, nanoprecipitation and solvent extraction/evaporation methods, were purpose-built in order to prepare FQPV-PLGA NPs. Both methods revealed suitable to obtain NPs with proper dimensions for the parenteral administration (< 250nm), narrow size distribution (P.I. about 0.1), good morphological features and negative charge (about -20mV). A peptide adsorption protocol onto NPs was considered as additional strategy to increase peptide expression on NPs surface aimed at improving the targeting effectiveness. A Design of Experiment approach (DoE) has been successfully applied to the more versatile solvent extraction/evaporation method in order to systematically highlight the influence of process parameters (organic solvent mixture, PVA concentration and polymeric solution volume) on NPs sizes.Conclusion: PLGA was successfully functionalized with two different peptides, FQPV and GE11, following the a versatile and simple carbodiimmide chemistry without further modifying polymer and/or peptide structure. Both nanoprecipitation and solvent extraction/evaporation NPs preparation methods were properly optimized in order to obtain peptide-PLGA based NPs and they can be alternatively selected according to solubility properties of both peptide-polymer conjugate and drug intended for encapsulation. Peptide adsorption on preformed PLGA NPs could be efficiently used to increase peptide expression on NPs surface thus improving cellular recognition in case of active targeting. Preliminary cytocompatibility evaluation of the selected peptide-PLGA based nanoparticulate materials shows a potential feasibility of the set-up synthetic procedures and NPs preparation methods for pharmaceutical purposes.