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Inhibition of adenosine kinase by phosphonate and bisphosphonate derivatives

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Abstract

The enzyme adenosine kinase (AK) plays a central role in regulating the intracellular and interstitial concentration of the purine nucleoside adenosine (Ado). In view of the beneficial effects of Ado in protecting tissues from ischemia and other stresses, there is much interest in developing AK inhibitors, which can regulate Ado concentration in a site- and event-specific manner. The catalytic activity of AK from different sources is dependent upon the presence of activators such as phosphate (Pi). In this work we describe several new phosphorylated compounds which either activate or inhibit AK. The compounds acetyl phosphate, carbamoyl phosphate, dihydroxyacetone phosphate and imidodiphosphate were found to stimulate AK activity in a dose-dependent manner comparable to that seen with Pi. In contrast, a number of phosphonate and bisphosphonate derivatives, which included clodronate and etidronate, were found to inhibit the activity of purified AK in the presence of Pi. These AK inhibitors (viz. clodronate, etidronate, phosphonoacetic acid, 2-carboxyethylphosphonic acid, N-(phosphonomethyl)-glycine and N-(phosphonomethyl)iminodiacetic acid), at concentrations at which they inhibited AK, were also shown to inhibit the uptake of 3H-adenosine and its incorporation into macromolecules in cultured mammalian cells, indicating that they were also inhibiting AK in intact cells. The drug concentrations at which these effects were observed showed limited toxicity to the cultured cells, indicating that these effects are not caused by cellular toxicity. These results indicate that the enzyme AK provides an additional cellular target for the clinically widely used bisphosphonates and related compounds, which could possibly be exploited for a new therapeutic application. Our structure–activity studies on different AK activators and inhibitors also indicate that all of the AK activating compounds have a higher partial positive charge (δ+) on the central phosphorous atom in comparison to the inhibitors. This information should prove helpful in the design and synthesis of more potent inhibitors of AK.

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Abbreviations

Ado:

adenosine

AK:

adenosine kinase

ADA:

adenosine deaminase

EHNA:

erythro-9-(2-hydroxyl-3-nonyl) adenine

AP:

acetyl phosphate

CP:

carbamoyl phosphate

DHAP:

dihydroxyacetone phosphate

IDP:

imidodiphosphate

MPA:

methylphosphonic acid

DEPAA:

diethylphosphonoacetic acid

PAA:

phosphonoacetic acid

PFA:

phosphonoformic acid

CEPA:

2-carboxyethylphosphonic acid

PMG:

N-(phosphonomethyl)-glycine

PMIA:

N-(phosphonomethyl)iminodiacetic acid

AR:

adenosine receptor

5′NH2dAdo:

5′-amino, 5′-deoxyadenosine

5IT:

5-iodotubercidin

5′d-5IT:

5′-deoxy, 5-iodotubercidin

MOPAC:

molecular orbital package

MNDO:

modified neglect of diatomic overlap

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  1. Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ontario, Canada, L8N 3Z5

    Jae Park, Bhag Singh & Radhey S. Gupta

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  1. Jae Park
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  2. Bhag Singh
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Correspondence to Radhey S. Gupta.

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Park, J., Singh, B. & Gupta, R.S. Inhibition of adenosine kinase by phosphonate and bisphosphonate derivatives. Mol Cell Biochem 283, 11–21 (2006). https://doi.org/10.1007/s11010-006-2216-7

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