The molecular mechanism of nitrogen-containing bisphosphonates as antiosteoporosis drugs

K. L. Kavanagh, K. Guo, James Edward Dunford, X. Wu, S. Knapp, F. H. Ebetino, Michael John Rogers, R. G. G. Russell, U. Oppermann

Research output: Contribution to journalArticle

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Abstract

Osteoporosis and low bone mass are currently estimated to be a major public health risk affecting > 50% of the female population over the age of 50. Because of their bone-selective pharmacokinetics, nitrogen-containing bisphosphonates (N-BPs), currently used as clinical inhibitors of bone-resorption diseases, target osteoclast farnesyl pyrophosphate synthase (FIRPS) and inhibit protein prenylation. FIRPS, a key branchpoint of the mevalonate pathway, catalyzes the successive condensation of isopentenyl pyrophosphate with dimethylallyl pyrophosphate and geranyl pyrophosphate. To understand the molecular events involved in inhibition of FPPS by N-BPs, we used protein crystallography, enzyme kinetics, and isothermal titration calorimetry. We report here high-resolution x-ray structures of the human enzyme in complexes with risedronate and zoledronate, two of the leading N-BPs in clinical use. These agents bind to the dimethylallyl/ geranyl pyrophosphate ligand pocket and induce a conformational change. The interactions of the N-BP cyclic nitrogen with Thr-201 and Lys-200 suggest that these inhibitors achieve potency by positioning their nitrogen in the proposed carbocation-binding site. Kinetic analyses reveal that inhibition is competitive with geranyl pyrophosphate and is of a slow, tight binding character, indicating that isomerization of an initial enzyme-inhibitor complex occurs with inhibitor binding. Isothermal titration calorimetry indicates that binding of N-BPs to the apoenzyme is entropy-driven, presumably through desolvation entropy effects. These experiments reveal the molecular binding characteristics of an important pharmacological target and provide a route for further optimization of these important drugs.

Original languageEnglish
Pages (from-to)7829-7834
Number of pages5
JournalPNAS
Volume103
Issue number20
DOIs
Publication statusPublished - May 2006

Keywords

  • farnesyl pyrophosphate synthase
  • osteoclast
  • slow, tight inhibition
  • farnesyl diphosphate synthase
  • trans-prenyltransferase
  • bone-resorption
  • in-vivo
  • mevalonate pathway
  • liver prenyltransferase
  • isoprenoid biosynthesis
  • pyrophosphate synthase
  • binding energetics
  • structural basis
  • inhibition

Cite this

Kavanagh, K. L., Guo, K., Dunford, J. E., Wu, X., Knapp, S., Ebetino, F. H., ... Oppermann, U. (2006). The molecular mechanism of nitrogen-containing bisphosphonates as antiosteoporosis drugs. PNAS, 103(20), 7829-7834. https://doi.org/10.1073/PNAS.0601643103

The molecular mechanism of nitrogen-containing bisphosphonates as antiosteoporosis drugs. / Kavanagh, K. L.; Guo, K.; Dunford, James Edward; Wu, X.; Knapp, S.; Ebetino, F. H.; Rogers, Michael John; Russell, R. G. G.; Oppermann, U.

In: PNAS, Vol. 103, No. 20, 05.2006, p. 7829-7834.

Research output: Contribution to journalArticle

Kavanagh, KL, Guo, K, Dunford, JE, Wu, X, Knapp, S, Ebetino, FH, Rogers, MJ, Russell, RGG & Oppermann, U 2006, 'The molecular mechanism of nitrogen-containing bisphosphonates as antiosteoporosis drugs', PNAS, vol. 103, no. 20, pp. 7829-7834. https://doi.org/10.1073/PNAS.0601643103
Kavanagh KL, Guo K, Dunford JE, Wu X, Knapp S, Ebetino FH et al. The molecular mechanism of nitrogen-containing bisphosphonates as antiosteoporosis drugs. PNAS. 2006 May;103(20):7829-7834. https://doi.org/10.1073/PNAS.0601643103
Kavanagh, K. L. ; Guo, K. ; Dunford, James Edward ; Wu, X. ; Knapp, S. ; Ebetino, F. H. ; Rogers, Michael John ; Russell, R. G. G. ; Oppermann, U. / The molecular mechanism of nitrogen-containing bisphosphonates as antiosteoporosis drugs. In: PNAS. 2006 ; Vol. 103, No. 20. pp. 7829-7834.
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AU - Knapp, S.

AU - Ebetino, F. H.

AU - Rogers, Michael John

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AU - Oppermann, U.

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AB - Osteoporosis and low bone mass are currently estimated to be a major public health risk affecting > 50% of the female population over the age of 50. Because of their bone-selective pharmacokinetics, nitrogen-containing bisphosphonates (N-BPs), currently used as clinical inhibitors of bone-resorption diseases, target osteoclast farnesyl pyrophosphate synthase (FIRPS) and inhibit protein prenylation. FIRPS, a key branchpoint of the mevalonate pathway, catalyzes the successive condensation of isopentenyl pyrophosphate with dimethylallyl pyrophosphate and geranyl pyrophosphate. To understand the molecular events involved in inhibition of FPPS by N-BPs, we used protein crystallography, enzyme kinetics, and isothermal titration calorimetry. We report here high-resolution x-ray structures of the human enzyme in complexes with risedronate and zoledronate, two of the leading N-BPs in clinical use. These agents bind to the dimethylallyl/ geranyl pyrophosphate ligand pocket and induce a conformational change. The interactions of the N-BP cyclic nitrogen with Thr-201 and Lys-200 suggest that these inhibitors achieve potency by positioning their nitrogen in the proposed carbocation-binding site. Kinetic analyses reveal that inhibition is competitive with geranyl pyrophosphate and is of a slow, tight binding character, indicating that isomerization of an initial enzyme-inhibitor complex occurs with inhibitor binding. Isothermal titration calorimetry indicates that binding of N-BPs to the apoenzyme is entropy-driven, presumably through desolvation entropy effects. These experiments reveal the molecular binding characteristics of an important pharmacological target and provide a route for further optimization of these important drugs.

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KW - osteoclast

KW - slow, tight inhibition

KW - farnesyl diphosphate synthase

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KW - bone-resorption

KW - in-vivo

KW - mevalonate pathway

KW - liver prenyltransferase

KW - isoprenoid biosynthesis

KW - pyrophosphate synthase

KW - binding energetics

KW - structural basis

KW - inhibition

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DO - 10.1073/PNAS.0601643103

M3 - Article

VL - 103

SP - 7829

EP - 7834

JO - PNAS

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SN - 0027-8424

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