Nanomedicines for active targeting: Physico-chemical characterization of paclitaxel-loaded anti-HER2 immunonanoparticles and in vitro functional studies on target cells
Nanomedicines for active targeting: Physico-chemical characterization of paclitaxel-loaded anti-HER2 immunonanoparticles and in vitro functional studies on target cells
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DOI：
10.1016/j.ejps.2009.07.006
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发表时间：
2009-10-08
影响因子：
4.6
通讯作者：
Delie, F.
中科院分区：
文献类型：
--
作者：
Cirstoiu-Hapca, A.;Buchegger, F.;Delie, F.
用人源化抗HER2单抗(曲妥珠单抗)共价偶联制备紫杉醇(TX)抗HER2免疫纳米粒(NPS-TX-HER)。Herceptin(R)到TX负载的聚(DL-乳酸)纳米粒(NPs-TX)，用于主动靶向过表达HER2受体的肿瘤细胞。比较NPs-TX-HER和未负载免疫纳米粒(NPs-HER)的物理化学性质，以评估药物对抗HER2偶联到NP表面的影响。采用盐析法制备的载药纳米粒(NPs-TX)的粒径为171+/-22 nm(P.I.=0.1)，包封率为78+/-10%，载药量为7.8+/-0.8%(w/w)。然后将NPS-TX硫代化并与活化的抗HER2单抗偶联，得到237+/-43 nm的免疫纳米粒(P.I.=0.2)。用I-125标记的单抗在SKOV-3细胞上检测激活步骤对单抗免疫反应性的影响，磺基MBS功能化后抗HER2单抗的活性几乎不受影响。每个纳米颗粒结合了大约270个抗HER2单抗分子。NPS-TX-HER的Zeta电位为0.2+/-0.1 mV。载TX免疫纳米粒的理化性质与未载药的免疫纳米粒非常相似，说明药物的包封性并不影响单抗与纳米粒的偶联。制备过程中未见药物丢失。DSC分析表明，包裹的TX为无定形或无序结晶相。这些结果表明，TX被包裹在聚合物基质中，而不是吸附在纳米粒子的表面。对SKOV-3卵巢癌细胞的体外研究表明，与其他TX制剂相比，NPS TX-HER具有更强的细胞毒作用。结果表明，在1ngTx/mlNP-TX-HER中，药物孵育的细胞存活率(77.32+/-5.48%)低于NPs-TX、Mabthera(R)包被的免疫纳米粒、无关单抗(NPs-TX-RIT)(93.8+/-12%)和游离药物(92.3+/-9.3%)。(C)2009爱思唯尔B.V.保留所有权利。
Paclitaxel (Tx)-loaded anti-HER2 immunonanoparticles (NPs-Tx-HER) were prepared by the covalent coupling of humanized monoclonal anti-HER2 antibodies (trastuzumab. Herceptin (R)) to Tx-loaded poly (DL-lactic acid) nanoparticles (NPs-Tx) for the active targeting of tumor cells that overexpress HER2 receptors. The physico-chemical properties of NPs-Tx-HER were compared to unloaded immunonanoparticles (NPs-HER) to assess the influence of the drug on anti-HER2 coupling to the NP surface. The immunoreactivity of sulfo-MBS activated anti-HER2 mAbs and the in vitro efficacy of NPs-Tx-HER were tested on SKOV-3 ovarian cancer cells that overexpress HER2 antigens.Tx-loaded nanoparticles (NPs-Tx) obtained by a salting-out method had a size of 171 +/- 22 nm (P.I.=0.1) and an encapsulation efficiency of about of 78 +/- 10%, which corresponded to a drug loading of 7.8 +/- 0.8% (w/w). NPs-Tx were then thiolated and conjugated to activated anti-HER2 mAbs to obtain immunonanoparticles of 237 +/- 43 nm (P.I.=0.2). The influence of the activation step on the immunoreactivity of the mAbs was tested on SKOV-3 cells using I-125-radiolabeled mAbs, and the activity of the anti-HER2 mAbs was minimally affected after sulfo-MBS functionalization. Approximately 270 molecules of anti-HER2 mAbs were bound per nanoparticle. NPs-Tx-HER exhibited a zeta potential of 0.2 +/- 0.1 mV. The physico-chemical properties of the Tx-loaded immunonanoparticles were very similar to unloaded immunonanoparticles, suggesting that the encapsulation of the drug did not influence the coupling of the mAbs to the NPs. No drug loss was observed during the preparation process. DSC analysis showed that encapsulated Tx is in an amorphous or disordered-crystalline phase. These results suggest that Tx is entrapped in the polymeric matrix and not adsorbed to the surface of the NPs. In vitro studies on SKOV-3 ovarian cancer cells demonstrated the greater cytotoxic effect of NPs Tx-HER compared to other Tx formulations. The results showed that at 1 ngTx/ml, the viability of cells incubated with drug encapsulated in NP-Tx-HER was lower (77.32 +/- 5.48%) than the viability of cells incubated in NPs-Tx (97.4 +/- 12%), immunonanoparticles coated with Mabthera (R), as irrelevant mAb (NPs-Tx-RIT) (93.8 +/- 12%) or free drug (92.3 +/- 9.3%). (C) 2009 Elsevier B.V. All rights reserved.