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顺铂生物缀合酶 GNPs 放大了顺铂的作用。

Cisplatin bioconjugated enzymatic GNPs amplify the effect of cisplatin with acquiescence.

机构信息

Nanomedicine & Nanobiotechnology Lab, Department of Biosciences, Integral University, Lucknow, India.

Department Pharmaceutics, College of Pharmacy, King Saud University, Riyadh, 11451, Saudi Arabia.

出版信息

Sci Rep. 2019 Sep 25;9(1):13826. doi: 10.1038/s41598-019-50215-y.

Abstract

Enzymatic gold nanoparticles (B-GNPs) have been synthesized using a natural anticancer agent bromelain (a cysteine protease) and these nanoparticles were used to bioconjugate Cisplatin (highly effective against osteosarcoma and lung cancer). Cisplatin bioconjugated bromelain encapsulated gold nanoparticles (B-C-GNPs) were found profoundly potent against same cancers at much lower concentration with minimum side effects due to the synergistic effect of bromelain. The B-C-GNPs have been observed to inhibit the proliferation of osteosarcoma cell lines Saos-2 and MG-63 with IC estimation of 4.51 µg/ml and 3.21 µg/ml, respectively, and against small lung cancer cell line A-549 with IC 2.5 µg/ml which is lower than IC of cisplatin against same cell lines. The B-GNPs/B-C-GNPs were characterized by TEM, UV-Visible spectroscopy, Zeta potential and DLS to confirm the production, purity, crystalline nature, stability of nanoemulsion, size and shape distribution. The change in 2D and 3D conformation of bromelain after encapsulation was studied by Circular Dichroism and Fluorometry, respectively. It was found that after encapsulation, a 19.4% loss in secondary structure was observed, but tertiary structure was not altered significantly and this loss improved the anticancer activity. The confirmation of bioconjugation of cisplatin with B-GNPs was done by UV-Visible spectroscopy, TEM, FTIR, 2D H NMR DOSY and ICP-MS. Further, it was found that almost ~4 cisplatin molecules bound with each B-GNPs nanoparticle.

摘要

酶促金纳米粒子(B-GNPs)已通过天然抗癌剂菠萝蛋白酶(半胱氨酸蛋白酶)合成,并用这些纳米粒子进行顺铂的生物偶联(对骨肉瘤和肺癌非常有效)。发现顺铂生物偶联菠萝蛋白酶包封金纳米粒子(B-C-GNPs)在低得多的浓度下对相同的癌症具有非常有效的作用,并且由于菠萝蛋白酶的协同作用,副作用最小。观察到 B-C-GNPs 抑制骨肉瘤细胞系 Saos-2 和 MG-63 的增殖,IC 估计值分别为 4.51μg/ml 和 3.21μg/ml,对小肺癌细胞系 A-549 的 IC 为 2.5μg/ml,低于顺铂对相同细胞系的 IC。通过 TEM、UV-可见光谱、Zeta 电位和 DLS 对 B-GNPs/B-C-GNPs 进行了表征,以确认生产、纯度、结晶性质、纳米乳液的稳定性、尺寸和形状分布。通过圆二色性和荧光光谱法分别研究了封装后菠萝蛋白酶的 2D 和 3D 构象变化。发现封装后,二级结构损失了 19.4%,但三级结构没有明显改变,这种损失提高了抗癌活性。通过 UV-可见光谱、TEM、FTIR、2D H NMR DOSY 和 ICP-MS 证实了顺铂与 B-GNPs 的生物偶联。此外,发现每个 B-GNPs 纳米粒子上几乎结合了~4 个顺铂分子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8315/6761153/ba9f33a4f440/41598_2019_50215_Fig1_HTML.jpg

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