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Amino Acids

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Development and validation of a sensitive LC–MS/MS assay for the quantification of anserine in human plasma and urine and its application to pharmacokinetic study

  • Inge Everaert
  • Giovanna Baron
  • Silvia Barbaresi
  • Ettore Gilardoni
  • Crescenzo Coppa
  • Marina Carini
  • Giulio Vistoli
  • Tine Bex
  • Jan Stautemas
  • Laura Blancquaert
  • Wim Derave
  • Giancarlo Aldini
  • Luca Regazzoni
Original Article
  • 38 Downloads
Part of the following topical collections:
  1. Carnosine

Abstract

Carnosine (beta-alanyl-l-histidine) and its methylated analogue anserine are present in relevant concentrations in the omnivore human diet. Several studies reported promising therapeutic potential for carnosine in various rodent models of oxidative stress and inflammation-related chronic diseases. Nevertheless, the poor serum stability of carnosine in humans makes the translation of rodent models hard. Even though anserine and carnosine have similar biochemical properties, anserine has better serum stability. Despite this interesting profile, the research on anserine is scarce. The aim of this study was to explore the bioavailability and stability of synthesized anserine by (1) performing in vitro stability experiments in human plasma and molecular modelling studies and by (2) evaluating the plasma and urinary pharmacokinetic profile in healthy volunteers following different doses of anserine (4–10–20 mg/kg body weight). A bio-analytical method for measuring anserine levels was developed and validated using liquid chromatography-electrospray mass spectrometry. Both plasma (CMAX: 0.54–1.10–3.12 µM) and urinary (CMAX: 0.09–0.41–0.72 mg/mg creatinine) anserine increased dose-dependently following ingestion of 4–10–20 anserine mg/kg BW, respectively. The inter-individual variation in plasma anserine was mainly explained by the activity (R2 = 0.75) and content (R2 = 0.77) of the enzyme serum carnosinase-1. Compared to carnosine, a lower interaction energy of anserine with carnosinase-1 was suggested by molecular modelling studies. Conversely, the two dipeptides seems to have similar interaction with the PEPT1 transporter. It can be concluded that nutritionally relevant doses of synthesized anserine are well-absorbed and that its degradation by serum carnosinase-1 is less pronounced compared to carnosine. This makes anserine a good candidate as a more stable carnosine-analogue to attenuate chronic diseases in humans.

Keywords

Anserine Carnosine Pharmacokinetics Carnosinase-1 

Notes

Acknowledgements

The practical contribution of Anneke Volkaert is greatly acknowledged. The authors thank Flamma (Italy) for generously providing anserine and carnosine. This work was financially supported by the Research Foundation-Flanders (FWO) Grant nos. (12R3815N, G035213N), and by the Industrial Research Fund (IOF, Ghent University) Grant F2014/IOF-StarTT/273. Inge Everaert is a recipient of a post-doc Fellowship by Research Foundation Flanders (FWO).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Research involving human participants

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

726_2018_2663_MOESM1_ESM.docx (83 kb)
Supplementary material 1 (DOCX 82 kb)

References

  1. Adelmann K, Frey D, Riedl E et al (2012) Different conformational forms of serum carnosinase detected by a newly developed sandwich ELISA for the measurements of carnosinase concentrations. Amino Acids 43:143–151.  https://doi.org/10.1007/s00726-012-1244-8 CrossRefPubMedGoogle Scholar
  2. Aldini G, Orioli M, Rossoni G et al (2011) The carbonyl scavenger carnosine ameliorates dyslipidemia and renal function in zucker obese rats. J Cell Mol Med 15:1339–1354CrossRefGoogle Scholar
  3. Bae ON, Serfozo K, Baek SH et al (2013) Safety and efficacy evaluation of carnosine, an endogenous neuroprotective agent for ischemic stroke. Stroke 44:205–212.  https://doi.org/10.1161/STROKEAHA.112.673954 CrossRefPubMedGoogle Scholar
  4. Baguet A, Everaert I, Yard B et al (2014) Does low serum carnosinase activity favor high-intensity exercise capacity? J Appl Physiol 116:553–559.  https://doi.org/10.1152/japplphysiol.01218.2013 CrossRefPubMedGoogle Scholar
  5. Barski OA, Xie Z, Baba SP et al (2013) Dietary carnosine prevents early atherosclerotic lesion formation in apolipoprotein e-null mice. Arterioscler Thromb Vasc Biol 33:1162–1170.  https://doi.org/10.1161/ATVBAHA.112.300572 CrossRefPubMedGoogle Scholar
  6. Boldyrev AA, Aldini G, Derave W (2013) Physiology and pathophysiology of carnosine. Physiol Rev 93:1803–1845.  https://doi.org/10.1152/physrev.00039.2012 CrossRefPubMedGoogle Scholar
  7. De Courten B, Jakubova M, De Courten MPJ et al (2016) Effects of carnosine supplementation on glucose metabolism: Pilot clinical trial. Obesity 24:1027–1034.  https://doi.org/10.1002/oby.21434 CrossRefPubMedGoogle Scholar
  8. Elbarbary NS, Ismail EAR, El-Naggar AR et al (2017) The effect of 12 weeks carnosine supplementation on renal functional integrity and oxidative stress in pediatric patients with diabetic nephropathy: a randomized placebo-controlled trial. Pediatr Diabetes.  https://doi.org/10.1111/pedi.12564 CrossRefPubMedGoogle Scholar
  9. Everaert I, Taes Y, De Heer E et al (2012) Low plasma carnosinase activity promotes carnosinemia after carnosine ingestion in humans. Am J Physiol Ren Physiol 302:F1537–F1544.  https://doi.org/10.1152/ajprenal.00084.2012 CrossRefGoogle Scholar
  10. Freedman BI, Hicks PJ, Sale MM et al (2007) A leucine repeat in the carnosinase gene CNDP1 is associated with diabetic end-stage renal disease in European Americans. Nephrol Dial Transplant 22:1131–1135.  https://doi.org/10.1093/ndt/gfl717 CrossRefPubMedGoogle Scholar
  11. Gardner M, Illingworth K, Wood DIANA (1991) Intestinal absorption of the intact peptide carnosine in man, and comparison with intestinal permeability of lactulose. J Physiol 439:411–422CrossRefGoogle Scholar
  12. Geissler S, Zwarg M, Knütter I et al (2010) The bioactive dipeptide anserine is transported by human proton-coupled peptide transporters. FEBS J 277:790–795.  https://doi.org/10.1111/j.1742-4658.2009.07528.x CrossRefPubMedGoogle Scholar
  13. Hajizadeh-Zaker R, Ghajar A, Mesgarpour B et al (2018) l-Carnosine As an Adjunctive Therapy to Risperidone in Children with Autistic Disorder: A Randomized, Double-Blind, Placebo-Controlled Trial. J Child Adolesc Psychopharmacol 28:74–81.  https://doi.org/10.1089/cap.2017.0026 CrossRefPubMedGoogle Scholar
  14. Harris RC, Tallon MJ, Dunnett M et al (2006) The absorption of orally supplied beta-alanine and its effect on muscle carnosine synthesis in human vastus lateralis. Amino Acids 30:279–289CrossRefGoogle Scholar
  15. Hirohiko M, Kazushige G, Toshitsugu Y et al (2006) Effects of carnosine and anserine supplementation on relatively high intensity endurance. J Biol Chem 4:86–94Google Scholar
  16. Hisatsune T, Kaneko J, Kurashige H et al (2015) Effect of anserine/carnosine supplementation on verbal episodic memory in elderly people. J Alzheimer’s Dis 50:149–159.  https://doi.org/10.3233/JAD-150767 CrossRefGoogle Scholar
  17. Houjeghani S, Kheirouri S, Faraji E, Jafarabadi MA (2018) L-Carnosine supplementation attenuated fasting glucose, triglycerides, advanced glycation end products, and tumor necrosis factor–α levels in patients with type 2 diabetes: a double-blind placebo-controlled randomized clinical trial. Nutr Res 49:96–106.  https://doi.org/10.1016/j.nutres.2017.11.003 CrossRefPubMedGoogle Scholar
  18. Iacobini C, Menini S, Blasetti Fantauzzi C et al (2018) FL-926-16, a novel bioavailable carnosinase-resistant carnosine derivative, prevents onset and stops progression of diabetic nephropathy in db/db mice. Br J Pharmacol 175:53–66.  https://doi.org/10.1111/bph.14070 CrossRefPubMedGoogle Scholar
  19. Janssen B, Hohenadel D, Brinkkoetter P et al (2005) Carnosine as a protective factor in diabetic nephropathy: association with a leucine repeat of the carnosinase gene CNDP1. Diabetes 54:2320–2327CrossRefGoogle Scholar
  20. Katakura Y, Totsuka M, Imabayashi E et al (2017) Anserine/carnosine supplementation suppresses the expression of the inflammatory chemokine CCL24 in peripheral blood mononuclear cells from elderly people. Nutrients 9:1199.  https://doi.org/10.3390/nu9111199 CrossRefPubMedCentralGoogle Scholar
  21. Konagai C, Watanabe H, Abe K et al (2013) Effects of essence of chicken on cognitive brain function: a near-infrared spectroscopy study. Biosci Biotechnol Biochem 77:178–181.  https://doi.org/10.1271/bbb.120706 CrossRefPubMedGoogle Scholar
  22. Kubomura D, Matahira Y, Masui A, Matsuda H (2009) Intestinal absorption and blood clearance of l-histidine-related compounds after ingestion of anserine in humans and comparison to anserine-containing diets. J Agric Food Chem 57:1781–1785.  https://doi.org/10.1021/jf8030875 CrossRefPubMedGoogle Scholar
  23. Matsumura Y, Kita S, Ono H et al (2002) Preventive effect of a chicken extract on the development of hypertension in stroke-prone spontaneously hypertensive rats. Biosci Biotechnol Biochem 66:1108–1110.  https://doi.org/10.1271/bbb.66.1108 CrossRefPubMedGoogle Scholar
  24. Mehrazad-Saber Z, Kheirouri S, Noorazar S-G (2018) Effects of L- Carnosine Supplementation on Sleep Disorders and Disease Severity in Autistic Children: A Randomized, Controlled Clinical Trial. Basic Clin Pharmacol Toxicol.  https://doi.org/10.1111/bcpt.12979 CrossRefGoogle Scholar
  25. Menini S, Iacobini C, Ricci C et al (2015) Protection from diabetes-induced atherosclerosis and renal disease by d-carnosine-octylester: effects of early vs late inhibition of advanced glycation end-products in Apoe-null mice. Diabetologia 58:845–853.  https://doi.org/10.1007/s00125-014-3467-6 CrossRefPubMedGoogle Scholar
  26. Mooyaart AL, Zutinic A, Bakker SJ et al (2010) Association between CNDP1 genotype and diabetic nephropathy is sex specific. Diabetes 59:1555–1559CrossRefGoogle Scholar
  27. Orioli M, Aldini G, Benfatto MC et al (2007) HNE Michael adducts to histidine and histidine-containing peptides as biomarkers of lipid-derived carbonyl stress in urines: LC-MS/MS profiling in Zucker obese rats. Anal Chem 79:9174–9184.  https://doi.org/10.1021/ac7016184 CrossRefPubMedGoogle Scholar
  28. Pedretti A, De Luca L, Marconi C et al (2008) Modeling of the intestinal peptide transporter hPepT1 and analysis of its transport capacities by docking and pharmacophore mapping. Chem Med Chem 3:1913–1921.  https://doi.org/10.1002/cmdc.200800184 CrossRefPubMedGoogle Scholar
  29. Pegova A, Abe H, Boldyrev A (2000) Hydrolysis of carnosine and related compounds by mammalian carnosinases. Comp Biochem Physiol B Biochem Mol Biol 127:443–446.  https://doi.org/10.1016/S0305-0491(00)00279-0 CrossRefPubMedGoogle Scholar
  30. Perry TL, Hansen S, Tischler B et al (1967) Carnosinemia: metabolic disorder with neurologic disease and mental defect. N Engl J Med 277:1219–1227CrossRefGoogle Scholar
  31. Peters V, Jansen EEW, Jakobs C et al (2011) Anserine inhibits carnosine degradation but in human serum carnosinase (CN1) is not correlated with histidine dipeptide concentration. Clin Chim Acta 412:263–267.  https://doi.org/10.1016/j.cca.2010.10.016 CrossRefPubMedGoogle Scholar
  32. Peters V, Schmitt CP, Weigand T et al (2017) Allosteric inhibition of carnosinase (CN1) by inducing a conformational shift. J Enzyme Inhib Med Chem 32:1102–1110.  https://doi.org/10.1080/14756366.2017.1355793 CrossRefPubMedPubMedCentralGoogle Scholar
  33. Sauerhofer S, Yuan G, Braun GS et al (2007) l-carnosine, a substrate of carnosinase-1, influences glucose metabolism. Diabetes 56:2425–2432CrossRefGoogle Scholar
  34. Stegen S, Stegen B, Aldini G et al (2015) Plasma carnosine, but not muscle carnosine, attenuates high-fat diet-induced metabolic stress. Appl Physiol Nutr Metab Physiol Appl Nutr Métabol 40:868–876.  https://doi.org/10.1139/apnm-2015-0042 CrossRefGoogle Scholar
  35. Suzuki Y (2004) Effect of CBEX supplementation on high-intensity intermittent exercise. Jpn J Phys Educ Health Sport Sci 49:159–169CrossRefGoogle Scholar
  36. Suzuki Y, Nakao T, Maemura H et al (2006) Carnosine and anserine ingestion enhances contribution of nonbicarbonate buffering. Med Sci Sports Exerc 38:334–338CrossRefGoogle Scholar
  37. Szcześniak D, Budzeń S, Kopeć W, Rymaszewska J (2014) Anserine and carnosine supplementation in the elderly: effects on cognitive functioning and physical capacity. Arch Gerontol Geriatr 59:485–490.  https://doi.org/10.1016/j.archger.2014.04.008 CrossRefPubMedGoogle Scholar
  38. Teufel M, Saudek V, Ledig JP et al (2003) Sequence identification and characterization of human carnosinase and a closely related non-specific dipeptidase. J Biol Chem 278:6521–6531CrossRefGoogle Scholar
  39. Vistoli G, Straniero V, Pedretti A et al (2012) Predicting the physicochemical profile of diastereoisomeric histidine-containing dipeptides by property space analysis. Chirality 24:566–576.  https://doi.org/10.1002/chir.22056 CrossRefPubMedGoogle Scholar
  40. Yadav A, Sinha N, Kumar V et al (2016) Association of CTG repeat polymorphism in carnosine dipeptidase 1 (CNDP1) gene with diabetic nephropathy in north Indians. Indian J Med Res 144:32.  https://doi.org/10.4103/0971-5916.193280 CrossRefPubMedPubMedCentralGoogle Scholar
  41. Yeum KJ, Orioli M, Regazzoni L et al (2010) Profiling histidine dipeptides in plasma and urine after ingesting beef, chicken or chicken broth in humans. Amino Acids 38:847–858.  https://doi.org/10.1007/s00726-009-0291-2 CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag GmbH Austria, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Department of Movement and Sports SciencesGhent UniversityGhentBelgium
  2. 2.Department of Pharmaceutical SciencesUniversity of MilanMilanItaly

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