Further insight into mechanism of action of clodronate: Inhibition of mitochondrial ADP/ATP translocase by a nonhydrolyzable, adenine-containing metabolite

P P Lehenkari, M Kellinsalmi, J P Napankangas, K V Ylitalo, J Monkkonen, M J Rogers, A Azhayev, H K Vaananen, I E Hassinen

Research output: Contribution to journalArticle

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Abstract

Bisphosphonates are currently the most important class of antiresorptive drugs used for the treatment of diseases with excess bone resorption. Recent studies have shown that bisphosphonates can be divided into two groups with distinct molecular mechanisms of action depending on the nature of the R 2 side chain. Alendronate, like other nitrogen-containing bisphosphonates, inhibits bone resorption and causes apoptosis of osteoclasts and other cells in vitro by preventing post-translational modification of GTP-binding proteins with isoprenoid lipids. Clodronate, a bisphosphonate that lacks a nitrogen, does not inhibit protein isoprenylation but can be metabolized intracellularly to a beta-gamma-methylene (AppCp-type) analog of ATP, which is cytotoxic to macrophages in vitro. The detailed molecular basis for the cytotoxic effects of adenosine-5'-[ beta,gamma-dichloromethylene] triphosphate (AppCCl(2)p) has not been determined yet. We addressed this question by studying the effects of alendronate, clodronate, and the clodronate metabolite AppCCl(2)p on isolated mitochondria, mitochondrial fractions, and mitochondrial membrane potential in isolated human osteoclasts. We found that AppCCl(2)p inhibits mitochondrial oxygen consumption by a mechanism that involves competitive inhibition of the ADP/ATP translocase. Alendronate or the native form of clodronate did not have any immediate effect on mitochondria. However, longer treatment with liposome-encapsulated clodronate caused collapse of the mitochondrial membrane potential, although prominent apoptosis was a late event. Hence, inhibition of the ADP/ATP translocase by the metabolite AppCCl(2)p is a likely route by which clodronate causes osteoclast apoptosis and inhibits bone resorption.

Original languageEnglish
Pages (from-to)1255-1262
Number of pages8
JournalMolecular Pharmacology
Volume61
Publication statusPublished - 2002

Keywords

  • NITROGEN-CONTAINING BISPHOSPHONATES
  • MOLD DICTYOSTELIUM-DISCOIDEUM
  • OSTEOCLASTS IN-VITRO
  • BONE-RESORPTION
  • PERMEABILITY TRANSITION
  • NUCLEOTIDE TRANSLOCATOR
  • MEVALONATE PATHWAY
  • MACROPHAGE-LIKE
  • APOPTOSIS
  • CELLS

Cite this

Lehenkari, P. P., Kellinsalmi, M., Napankangas, J. P., Ylitalo, K. V., Monkkonen, J., Rogers, M. J., ... Hassinen, I. E. (2002). Further insight into mechanism of action of clodronate: Inhibition of mitochondrial ADP/ATP translocase by a nonhydrolyzable, adenine-containing metabolite. Molecular Pharmacology, 61, 1255-1262.

Further insight into mechanism of action of clodronate: Inhibition of mitochondrial ADP/ATP translocase by a nonhydrolyzable, adenine-containing metabolite. / Lehenkari, P P ; Kellinsalmi, M ; Napankangas, J P ; Ylitalo, K V ; Monkkonen, J ; Rogers, M J ; Azhayev, A ; Vaananen, H K ; Hassinen, I E .

In: Molecular Pharmacology, Vol. 61, 2002, p. 1255-1262.

Research output: Contribution to journalArticle

Lehenkari, PP, Kellinsalmi, M, Napankangas, JP, Ylitalo, KV, Monkkonen, J, Rogers, MJ, Azhayev, A, Vaananen, HK & Hassinen, IE 2002, 'Further insight into mechanism of action of clodronate: Inhibition of mitochondrial ADP/ATP translocase by a nonhydrolyzable, adenine-containing metabolite' Molecular Pharmacology, vol. 61, pp. 1255-1262.
Lehenkari, P P ; Kellinsalmi, M ; Napankangas, J P ; Ylitalo, K V ; Monkkonen, J ; Rogers, M J ; Azhayev, A ; Vaananen, H K ; Hassinen, I E . / Further insight into mechanism of action of clodronate: Inhibition of mitochondrial ADP/ATP translocase by a nonhydrolyzable, adenine-containing metabolite. In: Molecular Pharmacology. 2002 ; Vol. 61. pp. 1255-1262.
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abstract = "Bisphosphonates are currently the most important class of antiresorptive drugs used for the treatment of diseases with excess bone resorption. Recent studies have shown that bisphosphonates can be divided into two groups with distinct molecular mechanisms of action depending on the nature of the R 2 side chain. Alendronate, like other nitrogen-containing bisphosphonates, inhibits bone resorption and causes apoptosis of osteoclasts and other cells in vitro by preventing post-translational modification of GTP-binding proteins with isoprenoid lipids. Clodronate, a bisphosphonate that lacks a nitrogen, does not inhibit protein isoprenylation but can be metabolized intracellularly to a beta-gamma-methylene (AppCp-type) analog of ATP, which is cytotoxic to macrophages in vitro. The detailed molecular basis for the cytotoxic effects of adenosine-5'-[ beta,gamma-dichloromethylene] triphosphate (AppCCl(2)p) has not been determined yet. We addressed this question by studying the effects of alendronate, clodronate, and the clodronate metabolite AppCCl(2)p on isolated mitochondria, mitochondrial fractions, and mitochondrial membrane potential in isolated human osteoclasts. We found that AppCCl(2)p inhibits mitochondrial oxygen consumption by a mechanism that involves competitive inhibition of the ADP/ATP translocase. Alendronate or the native form of clodronate did not have any immediate effect on mitochondria. However, longer treatment with liposome-encapsulated clodronate caused collapse of the mitochondrial membrane potential, although prominent apoptosis was a late event. Hence, inhibition of the ADP/ATP translocase by the metabolite AppCCl(2)p is a likely route by which clodronate causes osteoclast apoptosis and inhibits bone resorption.",
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T1 - Further insight into mechanism of action of clodronate: Inhibition of mitochondrial ADP/ATP translocase by a nonhydrolyzable, adenine-containing metabolite

AU - Lehenkari, P P

AU - Kellinsalmi, M

AU - Napankangas, J P

AU - Ylitalo, K V

AU - Monkkonen, J

AU - Rogers, M J

AU - Azhayev, A

AU - Vaananen, H K

AU - Hassinen, I E

PY - 2002

Y1 - 2002

N2 - Bisphosphonates are currently the most important class of antiresorptive drugs used for the treatment of diseases with excess bone resorption. Recent studies have shown that bisphosphonates can be divided into two groups with distinct molecular mechanisms of action depending on the nature of the R 2 side chain. Alendronate, like other nitrogen-containing bisphosphonates, inhibits bone resorption and causes apoptosis of osteoclasts and other cells in vitro by preventing post-translational modification of GTP-binding proteins with isoprenoid lipids. Clodronate, a bisphosphonate that lacks a nitrogen, does not inhibit protein isoprenylation but can be metabolized intracellularly to a beta-gamma-methylene (AppCp-type) analog of ATP, which is cytotoxic to macrophages in vitro. The detailed molecular basis for the cytotoxic effects of adenosine-5'-[ beta,gamma-dichloromethylene] triphosphate (AppCCl(2)p) has not been determined yet. We addressed this question by studying the effects of alendronate, clodronate, and the clodronate metabolite AppCCl(2)p on isolated mitochondria, mitochondrial fractions, and mitochondrial membrane potential in isolated human osteoclasts. We found that AppCCl(2)p inhibits mitochondrial oxygen consumption by a mechanism that involves competitive inhibition of the ADP/ATP translocase. Alendronate or the native form of clodronate did not have any immediate effect on mitochondria. However, longer treatment with liposome-encapsulated clodronate caused collapse of the mitochondrial membrane potential, although prominent apoptosis was a late event. Hence, inhibition of the ADP/ATP translocase by the metabolite AppCCl(2)p is a likely route by which clodronate causes osteoclast apoptosis and inhibits bone resorption.

AB - Bisphosphonates are currently the most important class of antiresorptive drugs used for the treatment of diseases with excess bone resorption. Recent studies have shown that bisphosphonates can be divided into two groups with distinct molecular mechanisms of action depending on the nature of the R 2 side chain. Alendronate, like other nitrogen-containing bisphosphonates, inhibits bone resorption and causes apoptosis of osteoclasts and other cells in vitro by preventing post-translational modification of GTP-binding proteins with isoprenoid lipids. Clodronate, a bisphosphonate that lacks a nitrogen, does not inhibit protein isoprenylation but can be metabolized intracellularly to a beta-gamma-methylene (AppCp-type) analog of ATP, which is cytotoxic to macrophages in vitro. The detailed molecular basis for the cytotoxic effects of adenosine-5'-[ beta,gamma-dichloromethylene] triphosphate (AppCCl(2)p) has not been determined yet. We addressed this question by studying the effects of alendronate, clodronate, and the clodronate metabolite AppCCl(2)p on isolated mitochondria, mitochondrial fractions, and mitochondrial membrane potential in isolated human osteoclasts. We found that AppCCl(2)p inhibits mitochondrial oxygen consumption by a mechanism that involves competitive inhibition of the ADP/ATP translocase. Alendronate or the native form of clodronate did not have any immediate effect on mitochondria. However, longer treatment with liposome-encapsulated clodronate caused collapse of the mitochondrial membrane potential, although prominent apoptosis was a late event. Hence, inhibition of the ADP/ATP translocase by the metabolite AppCCl(2)p is a likely route by which clodronate causes osteoclast apoptosis and inhibits bone resorption.

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KW - MOLD DICTYOSTELIUM-DISCOIDEUM

KW - OSTEOCLASTS IN-VITRO

KW - BONE-RESORPTION

KW - PERMEABILITY TRANSITION

KW - NUCLEOTIDE TRANSLOCATOR

KW - MEVALONATE PATHWAY

KW - MACROPHAGE-LIKE

KW - APOPTOSIS

KW - CELLS

M3 - Article

VL - 61

SP - 1255

EP - 1262

JO - Molecular Pharmacology

JF - Molecular Pharmacology

SN - 0026-895X

ER -