Synergistic Anti Leukemia Effect of a Novel Hsp90 and a Pan Cyclin Dependent Kinase Inhibitors.

Ashraf N Abdalla,Ammar Bader,Alaa A.-M Abdel-Aziz,Nunziatina De Tommasi,Hamad M Alkahtani,Antonio Vassallo,Mohammed H Mukhtar,Akhmed Aslam,Adel S El-Azab,Ahmed M Gouda,Waleed H Malki,Mahmoud Zaki El-Readi,Mohamed E Abdallah

doi:10.3390/molecules25092220

Abstract

Acute myeloid leukemia (AML) is among the top four malignancies in Saudi nationals, and it is the top leukemia subtype worldwide. Resistance to available AML drugs requires the identification of new targets and agents. Hsp90 is one of the emerging important targets in AML, which has a central role in the regulation of apoptosis and cell proliferation through client proteins including the growth factor receptors and cyclin dependent kinases. The objective of the first part of this study is to investigate the putative Hsp90 inhibition activity of three novel previously synthesized quinazolines, which showed HL60 cytotoxicity and VEGFR2 and EGFR kinases inhibition activities. Using surface plasmon resonance, compound 1 (HAA2020) showed better Hsp90 inhibition compared to 17-AAG, and a docking study revealed that it fits nicely into the ATPase site. The objective of the second part is to maximize the anti-leukemic activity of HAA2020, which was combined with each of the eleven standard inhibitors. The best resulting synergistic effect in HL60 cells was with the pan cyclin-dependent kinases (CDK) inhibitor dinaciclib, using an MTT assay. Furthermore, the inhibiting effect of the Hsp90α gene by the combination of HAA2020 and dinaciclib was associated with increased caspase-7 and TNF-α, leading to apoptosis in HL60 cells. In addition, the combination upregulated p27 simultaneously with the inhibition of cyclinD3 and CDK2, leading to abolished HL60 proliferation and survival. The actions of HAA2020 propagated the apoptotic and cell cycle control properties of dinaciclib, showing the importance of co-targeting Hsp90 and CDK, which could lead to the better management of leukemia.

Highlights

According to the Saudi Cancer Registry (2014), leukemia ranked second and sixth among the top malignancies for Saudi males and females, respectively, while it is the top malignancy for bothSaudi sexes below 14 years of age [1]
The interaction between each of the three compounds (1, 4, and 5) and Hsp90α was investigated in this study by a surface plasmon resonance (SPR)-based binding assay [30,31,32,33,34]
As a result of fitting the relative sensorgrams to a single-site bimolecular interaction model, the thermodynamic parameters for the resulting complex formation were determined. This approach allowed the measurement of 51.0 ± 2.9 nM

Summary

Introduction

Saudi sexes below 14 years of age [1]. Acute myeloid leukemia (AML) ranked fourth, with 11.9% of the total leukemia cases in both Saudi sexes during 1999–2013. The most common subtype of AML in Saudis is M1, with myeloperoxidase, CD13, CD33 and CD117 as the most common reported antigens [2,3]. AML is considered the top leukemia subtype, with a higher incidence in elderly patients [4]. Reviewing some of the effective targets, FLT3 is regarded as one of the most important mutations in AML, which is prevalent in over a third of its cases [5]. Sorafenib, lestaurtinib and midostaurin are considered the first generation of FLT3 inhibitors, as they have shown a broader effect on kinases

Methods

Results

Conclusion