Clodronate and liposome-encapsulated clodronate are metabolized to a toxic ATP analog, adenosine 5'-(beta,gamma-dichloromethylene) triphosphate, by mammalian cells in vitro

Julie Clare Crockett, J Monkkonen, G M Blackburn, R G G Russell, Michael John Rogers

Research output: Contribution to journalArticle

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Abstract

Clodronate, alendronate, and other bisphosphonates are widely used in the treatment of bone diseases characterized by excessive osteoclastic bone resorption, The exact mechanisms of action of bisphosphonates have not been identified but may involve a toxic effect on mature osteoclasts due to the induction of apoptosis, Clodronate encapsulated in liposomes is also toxic to macrophages in vivo and may therefore be of use in the treatment of inflammatory diseases, It is generally believed that bisphosphonates are not metabolized. However, we have found that mammalian cells in vitro (murine J774 macrophage-like cells and human MG63 osteosarcoma cells) can metabolize clodronate (dichloromethylenebisphosphonate) to a nonhydrolyzable adenosine triphosphate (ATP) analog, adenosine 5'-(beta,gamma-dichloromethylene) triphosphate, which could be detected in cell extracts by using fast protein liquid chromatography, J774 cells could also metabolize liposome-encapsulated clodronate to the same ATP analog, Liposome-encapsulated adenosine 5'-(beta,gamma-dichloromethylene) triphosphate was more potent than liposome-encapsulated clodronate at reducing the viability of cultures of J774 cells and caused both necrotic and apoptotic cell death, Neither alendronate nor liposome-encapsulated alendronate were metabolized, These results demonstrate that the toxic effect of clodronate on J774 macrophages, and probably on osteoclasts, is due to the metabolism of clodronate to a nonhydrolyzable ATP analog, Alendronate appears to act by a different mechanism.

Original languageEnglish
Pages (from-to)1358-1367
Number of pages10
JournalJournal of Bone and Mineral Research
Volume12
Issue number9
DOIs
Publication statusPublished - Sep 1997

Keywords

  • mold dictyostelium-discoideum
  • mononuclear phagocyte system
  • acute-phase response
  • bone-resorption
  • dichloromethylene diphosphonate
  • in-vitro
  • free bisphosphonates
  • rat bone
  • osteoclasts
  • macrophages

Cite this

Clodronate and liposome-encapsulated clodronate are metabolized to a toxic ATP analog, adenosine 5'-(beta,gamma-dichloromethylene) triphosphate, by mammalian cells in vitro. / Crockett, Julie Clare; Monkkonen, J ; Blackburn, G M ; Russell, R G G ; Rogers, Michael John.

In: Journal of Bone and Mineral Research, Vol. 12, No. 9, 09.1997, p. 1358-1367.

Research output: Contribution to journalArticle

Crockett, Julie Clare ; Monkkonen, J ; Blackburn, G M ; Russell, R G G ; Rogers, Michael John. / Clodronate and liposome-encapsulated clodronate are metabolized to a toxic ATP analog, adenosine 5'-(beta,gamma-dichloromethylene) triphosphate, by mammalian cells in vitro. In: Journal of Bone and Mineral Research. 1997 ; Vol. 12, No. 9. pp. 1358-1367.
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abstract = "Clodronate, alendronate, and other bisphosphonates are widely used in the treatment of bone diseases characterized by excessive osteoclastic bone resorption, The exact mechanisms of action of bisphosphonates have not been identified but may involve a toxic effect on mature osteoclasts due to the induction of apoptosis, Clodronate encapsulated in liposomes is also toxic to macrophages in vivo and may therefore be of use in the treatment of inflammatory diseases, It is generally believed that bisphosphonates are not metabolized. However, we have found that mammalian cells in vitro (murine J774 macrophage-like cells and human MG63 osteosarcoma cells) can metabolize clodronate (dichloromethylenebisphosphonate) to a nonhydrolyzable adenosine triphosphate (ATP) analog, adenosine 5'-(beta,gamma-dichloromethylene) triphosphate, which could be detected in cell extracts by using fast protein liquid chromatography, J774 cells could also metabolize liposome-encapsulated clodronate to the same ATP analog, Liposome-encapsulated adenosine 5'-(beta,gamma-dichloromethylene) triphosphate was more potent than liposome-encapsulated clodronate at reducing the viability of cultures of J774 cells and caused both necrotic and apoptotic cell death, Neither alendronate nor liposome-encapsulated alendronate were metabolized, These results demonstrate that the toxic effect of clodronate on J774 macrophages, and probably on osteoclasts, is due to the metabolism of clodronate to a nonhydrolyzable ATP analog, Alendronate appears to act by a different mechanism.",
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T1 - Clodronate and liposome-encapsulated clodronate are metabolized to a toxic ATP analog, adenosine 5'-(beta,gamma-dichloromethylene) triphosphate, by mammalian cells in vitro

AU - Crockett, Julie Clare

AU - Monkkonen, J

AU - Blackburn, G M

AU - Russell, R G G

AU - Rogers, Michael John

PY - 1997/9

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N2 - Clodronate, alendronate, and other bisphosphonates are widely used in the treatment of bone diseases characterized by excessive osteoclastic bone resorption, The exact mechanisms of action of bisphosphonates have not been identified but may involve a toxic effect on mature osteoclasts due to the induction of apoptosis, Clodronate encapsulated in liposomes is also toxic to macrophages in vivo and may therefore be of use in the treatment of inflammatory diseases, It is generally believed that bisphosphonates are not metabolized. However, we have found that mammalian cells in vitro (murine J774 macrophage-like cells and human MG63 osteosarcoma cells) can metabolize clodronate (dichloromethylenebisphosphonate) to a nonhydrolyzable adenosine triphosphate (ATP) analog, adenosine 5'-(beta,gamma-dichloromethylene) triphosphate, which could be detected in cell extracts by using fast protein liquid chromatography, J774 cells could also metabolize liposome-encapsulated clodronate to the same ATP analog, Liposome-encapsulated adenosine 5'-(beta,gamma-dichloromethylene) triphosphate was more potent than liposome-encapsulated clodronate at reducing the viability of cultures of J774 cells and caused both necrotic and apoptotic cell death, Neither alendronate nor liposome-encapsulated alendronate were metabolized, These results demonstrate that the toxic effect of clodronate on J774 macrophages, and probably on osteoclasts, is due to the metabolism of clodronate to a nonhydrolyzable ATP analog, Alendronate appears to act by a different mechanism.

AB - Clodronate, alendronate, and other bisphosphonates are widely used in the treatment of bone diseases characterized by excessive osteoclastic bone resorption, The exact mechanisms of action of bisphosphonates have not been identified but may involve a toxic effect on mature osteoclasts due to the induction of apoptosis, Clodronate encapsulated in liposomes is also toxic to macrophages in vivo and may therefore be of use in the treatment of inflammatory diseases, It is generally believed that bisphosphonates are not metabolized. However, we have found that mammalian cells in vitro (murine J774 macrophage-like cells and human MG63 osteosarcoma cells) can metabolize clodronate (dichloromethylenebisphosphonate) to a nonhydrolyzable adenosine triphosphate (ATP) analog, adenosine 5'-(beta,gamma-dichloromethylene) triphosphate, which could be detected in cell extracts by using fast protein liquid chromatography, J774 cells could also metabolize liposome-encapsulated clodronate to the same ATP analog, Liposome-encapsulated adenosine 5'-(beta,gamma-dichloromethylene) triphosphate was more potent than liposome-encapsulated clodronate at reducing the viability of cultures of J774 cells and caused both necrotic and apoptotic cell death, Neither alendronate nor liposome-encapsulated alendronate were metabolized, These results demonstrate that the toxic effect of clodronate on J774 macrophages, and probably on osteoclasts, is due to the metabolism of clodronate to a nonhydrolyzable ATP analog, Alendronate appears to act by a different mechanism.

KW - mold dictyostelium-discoideum

KW - mononuclear phagocyte system

KW - acute-phase response

KW - bone-resorption

KW - dichloromethylene diphosphonate

KW - in-vitro

KW - free bisphosphonates

KW - rat bone

KW - osteoclasts

KW - macrophages

U2 - 10.1359/jbmr.1997.12.9.1358

DO - 10.1359/jbmr.1997.12.9.1358

M3 - Article

VL - 12

SP - 1358

EP - 1367

JO - Journal of Bone and Mineral Research

JF - Journal of Bone and Mineral Research

SN - 0884-0431

IS - 9

ER -