A computational and laboratory approach for the investigation of interactions of peptide conjugated natural terpenes with EpHA2 receptor
A computational and laboratory approach for the investigation of interactions of peptide conjugated natural terpenes with EpHA2 receptor
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DOI：
10.1007/s00894-023-05596-3
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发表时间：
2023-07-01
影响因子：
2.2
通讯作者：
Banerjee，Ipsita A. A.
中科院分区：
文献类型：
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作者：
Goncalves，Beatriz G. G.;Banerjee，Ipsita A. A.
Ephrin A型受体2（EphA 2）是一种众所周知的癌症治疗药物靶点，因为它在许多类型的癌症中过表达。因此，通过靶向方法确定该受体与配体结合结构域（LBD）和激酶结合结构域（KBD）的结合相互作用以调节其活性是至关重要的。在这项工作中，具有固有抗癌特性的天然萜烯与已知与EphA 2受体的LBD结合的短肽YSAYP和SWLAY缀合。我们研究了六个萜烯（山楂酸，左旋海松酸，quinopimaric acid，oleopentane，polyalthic，和hydroxybetulinic acid）共轭上述肽与配体结合结构域（LBD）的EphA 2受体的计算结合相互作用。此外，在“靶标跳跃方法”之后，我们还检查了缀合物与大骨节病的相互作用。我们的结果表明，与LBD相比，大多数缀合物显示出与EphA 2激酶结构域更高的结合相互作用。此外，萜烯的结合亲和力增加后，缀合的肽与萜烯。为了进一步研究对EphA 2激酶结构域的特异性，我们还检查了与VPWXE（x =正亮氨酸）缀合的萜烯的结合相互作用，因为VPWXE已显示与其他RTK结合。我们的研究结果表明，特别是与SWLAY缀合的萜类化合物对结合大骨节病表现出高效力。我们还设计了缀合物，其中肽部分和萜烯通过丁基（C4）基团接头分开，以检查结合相互作用是否可以增强。对接研究表明，与没有接头的缀合物相比，具有接头的缀合物与LBD的结合增强，尽管没有接头的缀合物与KBD的结合仍然略高。作为概念的证明，然后用已知过表达EphA 2受体的F98肿瘤细胞测试每种肽的maslinate和oleanolate缀合物。结果表明，齐墩果酸酯-酰胺基-SWLAY缀合物在降低肿瘤细胞的细胞增殖方面是有效的，并且可以潜在地开发和进一步研究用于靶向过表达EphA 2受体的肿瘤细胞。为了测试这些缀合物是否可以与受体结合并潜在地作为激酶抑制剂起作用，我们进行了SPR分析和ADP-Glo测定。我们的研究结果表明，OA与SWLAY共轭物表现出最高的inhibit.MethodsDocking研究进行了使用AutoDock维纳，v.1.2.0;分子动力学和MMGBSA计算进行了通过薛定谔软件DESMOND。
ContextEphrin type A receptor 2 (EphA2) is a well-known drug target for cancer treatment due to its overexpression in numerous types of cancers. Thus, it is crucial to determine the binding interactions of this receptor with both the ligand-binding domain (LBD) and the kinase-binding domain (KBD) through a targeted approach in order to modulate its activity. In this work, natural terpenes with inherent anticancer properties were conjugated with short peptides YSAYP and SWLAY that are known to bind to the LBD of EphA2 receptor. We examined the binding interactions of six terpenes (maslinic acid, levopimaric acid, quinopimaric acid, oleanolic, polyalthic, and hydroxybetulinic acid) conjugated to the above peptides with the ligand-binding domain (LBD) of EphA2 receptor computationally. Additionally, following the “target-hopping approach,” we also examined the interactions of the conjugates with the KBD. Our results indicated that most of the conjugates showed higher binding interactions with the EphA2 kinase domain compared to LBD. Furthermore, the binding affinities of the terpenes increased upon conjugating the peptides with the terpenes. In order to further investigate the specificity toward EphA2 kinase domain, we also examined the binding interactions of the terpenes conjugated to VPWXE (x = norleucine), as VPWXE has been shown to bind to other RTKs. Our results indicated that the terpenes conjugated to SWLAY in particular showed high efficacy toward binding to the KBD. We also designed conjugates where in the peptide portion and the terpenes were separated by a butyl (C4) group linker to examine if the binding interactions could be enhanced. Docking studies showed that the conjugates with linkers had enhanced binding with the LBD compared to those without linkers, though binding remained slightly higher without linkers toward the KBD. As a proof of concept, maslinate and oleanolate conjugates of each of the peptides were then tested with F98 tumor cells which are known to overexpress EphA2 receptor. Results indicated that the oleanolate-amido-SWLAY conjugates were efficacious in reducing the cell proliferation of the tumor cells and may be potentially developed and further studied for targeting tumor cells overexpressing the EphA2 receptor. To test if these conjugates could bind to the receptor and potentially function as kinase inhibitors, we conducted SPR analysis and ADP-Glo assay. Our results indicated that OA conjugate with SWLAY showed the highest inhibition.MethodsDocking studies were carried out using AutoDock Vina, v.1.2.0; Molecular Dynamics and MMGBSA calculations were carried out through Schrodinger Software DESMOND.