The EZMTT cell proliferation assay provides precise measurement for drug combinations and better correlation between in vitro and in vivo efficacy

Rui, Jingjing; Yin, JianBing; Yu, Yan; Bai, Ruisong; Su, Weike; Ruan, Benfang

doi:10.1016/j.bmcl.2020.127134

Cited by 2 publications

(4 citation statements)

References 25 publications

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“…In a 96-well plate, 4 × 10 3 /well of the cells were plated and treated with a series of dilutions of CPD 23 in PBS with 1% DMSO, CPD 23@SOL micelles (both CPD 23 topped at 2 mM) and blank SOL micelles for 5 days. The amount of NAD(P)H produced in viable cells was measured using the EZMTT reagents 35 , 36 with 1 hour incubation, to collect OD values of the samples, control and blank wells at the wavelength of 450 nm. The inhibition of specimens to tumor cells was calculated by equation (3).…”

Section: Materials and Experimental Methodsmentioning

confidence: 99%

Self-Assembled Micellar Glutaminase Allosteric Inhibitor for Effective Therapeutic Intervention

Fang¹,

Chen²,

Li³

et al. 2022

IJN

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Introduction: Kidney-type glutaminase (KGA) has been an important anti-tumor drug target, and KGA allosteric inhibitors attracted much interest for their superior enzymatic specificity with good drug safety profiles. For glutaminase allosteric inhibitors such as BPTES, CB-839 and Selen derivatives, the low solubility remains as the main factor that limits in vivo efficacy. The 1,3,4-Selenadiazole compound CPD 23 showed improved in vivo efficacy but worse solubility; however, the graft polymer polyvinyl caprolactam-polyvinyl acetate-polyethylene glycol (PVCap-PVA-PEG), Soluplus ® (SOL) stood out as an excellent delivery carrier for CPD 23. Methods: The CPD 23@SOL micelles were prepared, optimized and evaluated through on the basis of solubility improvement and loading capacity. Characterizations of particle size and Zeta potential by dynamic light scattering, morphology by transmission electron microscopy and solid state by X-ray powder diffraction were closely conducted. The biological studies included the tumor cell growth inhibition, blood and liver microsomal stability, in vivo pharmacokinetics and tissue biodistribution. Results: At 1:20 ratio of CPD 23:SOL, CPD 23@SOL micelles were well-dispersed, spherical and stable, with size less than 200 nm with encapsulation efficiency of more than 90%. This SOL micellar system significantly increased the aqueous solubility of CPD 23 by 15,000 folds. Particularly, CPD 23@SOL micelles demonstrated higher stability in blood and liver microsomes, showing approximately 86% remaining at 2 h incubation and about 66% at 4 h, respectively. In addition, with or without micellar formulation, CPD 23 maintained essentially the same inhibitory activity in tumor cells. Interestingly, CPD 23@SOL micelles significantly improved the pharmacokinetic exposure, prolonged the in vivo circulation and dramatically changed tissue biodistributions of CPD 23. Conclusion:The current work provided an encouraging and practical delivery system for novel Selenadiazoles and glutaminase allosteric inhibitors whose poor water-soluble characteristic has been a bottleneck for the field.

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Section: Materials and Experimental Methodsmentioning

confidence: 99%

Self-Assembled Micellar Glutaminase Allosteric Inhibitor for Effective Therapeutic Intervention

Fang¹,

Chen²,

Li³

et al. 2022

IJN

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“…However, clinical applications of glutaminase inhibitors have been met with major obstacles and generally poor outcomes. KGA allosteric inhibitors BPTES and CB839 showed partial inhibition of many cancer cell lines; the observed drug induced proliferation rate (DIP) for overnight culture containing CB839 was more than 10% of the one without drug treatment . Also, at high dosage (200 mg/kg oral), CB839 only achieved 50% tumor growth suppression in an in vivo xenograft model.…”

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confidence: 98%

“…KGA allosteric inhibitors BPTES and CB839 4 showed partial inhibition of many cancer cell lines; the observed drug induced proliferation rate (DIP) for overnight culture containing CB839 was more than 10% of the one without drug treatment. 5 Also, at high dosage (200 mg/kg oral), CB839 only achieved 50% tumor growth suppression in an in vivo xenograft model. Recently, Soth et al 6 developed a new compound IPN60090 that is currently in Clinical Phase I and showed high selectivity for GLS1 (IC 50 = 31 nM), but no activity against GLS2.…”

mentioning

confidence: 99%

“…The lack of in vivo efficacy by a single agent might be due to alternative pathways that could bypass the glutaminase inhibition. To overcome this, glutaminase inhibitors were tested in drug combination that showed synergy with paclitaxel and rapamycin in reducing tumor growth, indicating a suitable combination of a couple of targets might completely inhibit cancer growth and overcome drug resistance. Hexylselen , was designed as a dual KGA/GDH inhibitor, and it can completely inhibit many cancer cells with low toxicity to the normal cells.…”

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confidence: 99%

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Diselenide Covalent Allosteric Inhibitors of Glutaminase with Strong In Vivo Anticancer Activity

Hou

Chen

Pan

et al. 2023

ACS Med. Chem. Lett.

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Allosteric glutaminase inhibitors demonstrate inhibition of glutamine-dependent cancer cells with low general drug toxicity, but have issues with efficacy in vivo. Here, we designed a series of diselenide compounds with 6 atoms in the middle, aiming to target the allosteric site of kidney type glutaminase (KGA) with a covalent linkage to strengthen the interaction. Proteomic analysis demonstrated that the diselenide compounds cross-linked with the Lys320 residue at the KGA allosteric site; this was confirmed by the KGA K320A mutant which showed essentially no binding to the diselenide. Further, structure−activity relationship (SAR) analysis demonstrated that growth inhibition correlated well with KGA inhibition and was enhanced by thioredoxin reductase (TrxR) inhibition. Interestingly, diselenide compounds showed no inhibition of glutamate dehydrogenase (GDH), indicating some enzyme selectivity. Importantly, the designed novel diselenides are glutaminase allosteric inhibitors that showed in vivo efficacy and survival in the xenograft animal model.

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The EZMTT cell proliferation assay provides precise measurement for drug combinations and better correlation between in vitro and in vivo efficacy

Cited by 2 publications

References 25 publications

Self-Assembled Micellar Glutaminase Allosteric Inhibitor for Effective Therapeutic Intervention

Self-Assembled Micellar Glutaminase Allosteric Inhibitor for Effective Therapeutic Intervention

Diselenide Covalent Allosteric Inhibitors of Glutaminase with Strong In Vivo Anticancer Activity

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