Phosphocyclocreatine is the dominant form of cyclocreatine in control and creatine transporter deficiency patient fibroblasts

Pharmacol Res Perspect. 2019 Dec;7(6):e00525. doi: 10.1002/prp2.525.

Abstract

Creatine transporter deficiency (CTD) is a metabolic disorder resulting in cognitive, motor, and behavioral deficits. Cyclocreatine (cCr), a creatine analog, has been explored as a therapeutic strategy for the treatment of CTD. We developed a rapid, selective, and accurate HILIC ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method to simultaneously quantify the intracellular concentrations of cCr, creatine (Cr), creatine-d3 (Cr-d3), phosphocyclocreatine (pcCr), and phosphocreatine (pCr). Using HILIC-UPLC-MS/MS, we measured cCr and Cr-d3 uptake and their conversion to the phosphorylated forms in primary human control and CTD fibroblasts. Altogether, the data demonstrate that cCr enters cells and its dominant intracellular form is pcCr in both control and CTD patient cells. Therefore, cCr may replace creatine as a therapeutic strategy for the treatment of CTD.

Keywords: HILIC UPLC-MS/MS method; creatine; creatine transporter deficiency; cyclocreatine; phosphocreatine; phosphocyclocreatine.

Publication types

  • Research Support, N.I.H., Intramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Brain Diseases, Metabolic, Inborn / drug therapy*
  • Brain Diseases, Metabolic, Inborn / metabolism
  • Cells, Cultured
  • Chromatography, High Pressure Liquid / methods
  • Creatine / deficiency*
  • Creatine / metabolism
  • Creatinine / analogs & derivatives*
  • Creatinine / pharmacokinetics
  • Creatinine / therapeutic use
  • Fibroblasts / metabolism*
  • Humans
  • Imidazolidines / analysis
  • Imidazolidines / metabolism*
  • Mental Retardation, X-Linked / drug therapy*
  • Mental Retardation, X-Linked / metabolism
  • Phosphocreatine / analogs & derivatives*
  • Phosphocreatine / analysis
  • Phosphocreatine / metabolism
  • Plasma Membrane Neurotransmitter Transport Proteins / deficiency*
  • Plasma Membrane Neurotransmitter Transport Proteins / metabolism
  • Primary Cell Culture
  • Tandem Mass Spectrometry / methods

Substances

  • Imidazolidines
  • Plasma Membrane Neurotransmitter Transport Proteins
  • Phosphocreatine
  • phosphocyclocreatine
  • cyclocreatine
  • Creatinine
  • Creatine

Supplementary concepts

  • Creatine deficiency, X-linked