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Reversal of genetic brain iron accumulation by N,N′-bis(2-mercaptoethyl)isophthalamide, a lipophilic metal chelator, in mice

  • Inorganic Compounds
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Abstract

N,N’-bis(2-mercaptoethyl)isophthalamide (NBMI) is a novel lipophilic metal chelator and antioxidant used in mercury poisoning. Recent studies have suggested that NBMI may also bind to other metals such as lead and iron. Since NBMI can enter the brain, we evaluated if NBMI removes excess iron from the iron-loaded brain and ameliorates iron-induced oxidative stress. First, NBMI exhibited preferential binding to ferrous (Fe2+) iron with a negligible binding affinity to ferric (Fe3+) iron, indicating a selective chelation of labile iron. Second, NBMI protected SH-SY5Y human neuroblastoma cells from the cytotoxic effects of high iron. NBMI also decreased cellular labile iron and lessened the production of iron-induced reactive oxygen species in these cells. Deferiprone (DFP), a commonly used oral iron chelator, failed to prevent iron-induced cytotoxicity or labile iron accumulation. Next, we validated the efficacy of NBMI in Hfe H67D mutant mice, a mouse model of brain iron accumulation (BIA). Oral gavage of NBMI for 6 weeks decreased iron accumulation in the brain as well as liver, whereas DFP showed iron chelation only in the liver, but not in the brain. Notably, depletion of brain copper and anemia were observed in BIA mice treated with DFP, but not with NBMI, suggesting a superior safety profile of NBMI over DFP for long-term use. Collectively, our study demonstrates that NBMI provides a neuroprotective effect against BIA and has therapeutic potential for neurodegenerative diseases associated with BIA.

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Abbreviations

AD:

Alzheimer’s disease

ANOVA:

Analysis of variance

BBB:

Blood–brain barrier

BIA:

Brain iron accumulation

CMC:

Carboxymethylcellulose

DCF-DA:

2’,7’-Dichlorofluorescein diacetate

DFO:

Deferoxamine

DFP:

Deferiprone

DMEM:

Dulbecco’s modified eagle’s medium

DMPS:

Sodium 2,3-dimercaptopropanesulfonate monohydrate

DMSO:

Dimethyl sulfoxide

FAC:

Ferric ammonium citrate

FBS:

Fetal bovine serum

Fe2 + :

Ferrous iron

Fe3 + :

Ferric iron

FeCl3 :

Iron (III) chloride

Hg:

Mercury

MCH:

Mean corpuscular hemoglobin

MCV:

Mean corpuscular volume

MES:

2-(N-Morpholino) ethanesulfonic acid

NBMI:

N,N’-Bis(2-mercaptoethyl)isophthalamide

Pb:

Lead

PD:

Parkinson’s disease

RBC:

Red blood cell

ROS:

Reactive oxygen species

WBC:

White blood cell

WT:

Wild type

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Acknowledgements

The authors are grateful to EmeraMed (Nicholasville, KY) for providing NBMI and reagents, to Dr. Haley Boyd and Dr. Kuljeet Gugnani for helpful discussions, to Ms. Lingxue Zeng and Mr. Sonny Farfan for help during animal experiments, and to Ms. Analucia Gonzales Urday and Mr. Shrey Shah for quantification of calcein-AM images.

Funding

This work was supported in part by EmeraMed.

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Author notes

  1. Ruiying Cheng and Rajitha Gadde have contributed equally to this work.

Authors and Affiliations

  1. Department of Biomedical and Nutritional Sciences, University of Massachusetts Lowell, Lowell, MA, USA

    Ruiying Cheng & Jonghan Kim

  2. Department of Pharmaceutical Sciences, School of Pharmacy, Massachusetts College of Pharmacy and Health Sciences, Boston, MA, USA

    Rajitha Gadde, Neha Kulkarni & Swati Betharia

  3. Department of Pharmaceutical Sciences, Northeastern University, Boston, MA, USA

    Yingfang Fan, Nachiket Shevale & Jonghan Kim

  4. Gordon Center for Medical Imaging, Department of Radiology, Massachusetts General Hospital and Harvard Medical School, Boston, MA, USA

    Kai Bao & Hak Soo Choi

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Contributions

RC, RG, YF, SB, and JK designed research; RC, RG, YF, NK, NS, and KB performed research; RC, RG, YF, SB, and JK analyzed data; RC prepared the original draft; RG, SB, and JK reviewed and edited the manuscript; HSC, SB, and JK provided necessary lab space and reagents for research.

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Correspondence to Swati Betharia or Jonghan Kim.

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Cheng, R., Gadde, R., Fan, Y. et al. Reversal of genetic brain iron accumulation by N,N′-bis(2-mercaptoethyl)isophthalamide, a lipophilic metal chelator, in mice. Arch Toxicol 96, 1951–1962 (2022). https://doi.org/10.1007/s00204-022-03287-1

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