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Excretion, Metabolism and Cytochrome P450 Inhibition of Methyl 3,4-Dihydroxybenzoate (MDHB): A Potential Candidate to Treat Neurodegenerative Diseases

  • Jia Hui Wang
  • Ke Qi Chen
  • Jun Xing Jiang
  • Huan Yong Li
  • Jun Ping Pan
  • Ji Yan Su
  • Lin Wang
  • Yang Hu
  • Xiang Nan Mi
  • Yi Rong Xin
  • Qin Gao
  • Xiang Long Zhao
  • Fei XiaoEmail author
  • Huan Min LuoEmail author
Original Research Article
  • 25 Downloads

Abstract

Background and Objectives

Methyl 3,4-dihydroxybenzoate (MDHB) has the potential to prevent neurodegenerative diseases (NDDs). The present work investigated its excretion, metabolism, and cytochrome P450-based drug–drug interactions (DDIs).

Methods

After intragastric administration of MDHB, the parent drug was assayed in the urine and faeces of mice. Metabolites of MDHB in the urine, faeces, brain, plasma and liver were detected by liquid chromatography–hybrid quadrupole time-of-flight mass spectrometry (LC–QTOF/MS). A cocktail approach was used to evaluate the inhibition of cytochrome P450 isoforms by MDHB.

Results

The cumulative excretion permille of MDHB in the urine and faeces were found to be 0.67 ± 0.31 and 0.49 ± 0.44‰, respectively. A total of 96 metabolites of MDHB were identified, and all IC50 (half-maximal inhibitory concentration) values of MDHB towards cytochrome P450 isoforms were > 100 μM.

Conclusions

The results suggest that MDHB has a low parent drug cumulative excretion percentage and that MDHB has multiple metabolites and is mainly metabolized through the loss of –CH2 and –CO2, the loss of –CH2O, ester bond hydrolysis, the loss of –O and –CO2, isomerization, methylation, sulfate conjugation, the loss of –CH2O and –O and glycine conjugation, glycine conjugation, the loss of two –O groups and alanine conjugation, the loss of –CH2O and –O and glucose conjugation, glucuronidation, glucose conjugation, etc., in vivo. Finally, MDHB has a low probability of cytochrome P450-based DDIs.

Notes

Compliance with Ethical Standards

Ethical approval

The animal experiments adhered to the Jinan University Medical College Animal Use Ordinance and were approved by the Ethics Committee of the Medical School of Jinan University.

Funding

This work was supported by the National Natural Science Foundation of China (grant no. 81473296) and China’s 111 Project (no. B14036).

Conflict of interest

None of the authors have conflicts of interest to declare.

Supplementary material

13318_2019_576_MOESM1_ESM.docx (17.5 mb)
Supplementary material 1 (DOCX 17925 kb)

References

  1. 1.
    Ma L, Li JM, Chen YQ, Li Y, Guo XJ. Determination of 3,4-dihydroxy methyl benzoate in Hedyotis diffusa Willd by HPLC. Shizhen Guo Yi Guo Yao. 2009;20(03):528–9.Google Scholar
  2. 2.
    Xu WQ, Gong XJ, Zhou X, Zhao C, Chen HG. Chemiacal constituents and bioactivity of Kalimeris indica. China J Chin Materia Med. 2010;35(23):3172–4.Google Scholar
  3. 3.
    Zhang Z, Zhou X, Zhou XW, Xu X, Liao MJ, Yan L, et al. Methyl 3,4-dihydroxybenzoate promotes neurite outgrowth of cortical neurons cultured in vitro. Neural Regen Res. 2012;7(13):971–7.PubMedPubMedCentralGoogle Scholar
  4. 4.
    Zhang Z, Cai L, Zhou XW, Su CF, Xiao F, Gao Q, et al. Methyl 3,4-dihydroxybenzoate promote rat cortical neurons survival and neurite outgrowth through the adenosine A2a receptor/PI3K/Akt signaling pathway. NeuroReport. 2015;26(6):367–73.CrossRefGoogle Scholar
  5. 5.
    Zhou X, Su CF, Zhang Z, Wang CY, Luo JQ, Zhou XW, et al. Neuroprotective effects of methyl 3,4-dihydroxybenzoate against H2O2-induced apoptosis in RGC-5 cells. J Pharmacol Sci. 2014;125(1):51–8.CrossRefGoogle Scholar
  6. 6.
    Cai L, Wang LF, Pan JP, Mi XN, Zhang Z, Geng HJ, et al. Neuroprotective effects of methyl 3,4-dihydroxybenzoate against TBHP-induced oxidative damage in SH-SY5Y cells. Molecules. 2016;21(8):1071.CrossRefGoogle Scholar
  7. 7.
    Zhou XW, Zhang Z, Su CF. Methyl 3,4-dihydroxybenzoate protects primary cortical neurons against Aβ25-35-induced neurotoxicity through mitochondria pathway. J Neurosci Res. 2013;91:1215–25.CrossRefGoogle Scholar
  8. 8.
    Zhang J, Xu D, Ouyang H, Hu SH, Li A, Luo HM, et al. Neuroprotective effects of methyl 3,4-dihydroxybenzoate in a mouse model of retinitis pigmentosa. Exp Eye Res. 2017;162:86–96.CrossRefGoogle Scholar
  9. 9.
    Hu SH, Wang JH, Mi XN, Pan JP, Wang LF, Zhang J, et al. Neuroprotective effect of methyl 3,4-dihydroxybenzoate on NMDA-induced injury in rat retina. Int J Sci. 2018;5(5):114–29.Google Scholar
  10. 10.
    Zhang W, Cai L, Geng HJ, Su CF, Yan L, Wang JH, et al. Methyl 3,4-dihydroxybenzoate extends the lifespan of Caenorhabditis elegans, partly via W06A7.4 gene. Exp Gerontol. 2014;60:108–16.Google Scholar
  11. 11.
    Wang JH, Hu SH, Su JY, Pan JP, Mi XN, Geng HJ, et al. Determination of the pharmacokinetics and tissue distribution of methyl 3,4-dihydroxybenzoate (MDHB) in mice using liquid chromatography–tandem mass spectrometry. Eur J Drug Metab Pharmacokinet. 2019;44:237–49.CrossRefGoogle Scholar
  12. 12.
    Yang BF. Pharmacology, vol. 7. Beijing: People’s Medical Publishing House; 2008.Google Scholar
  13. 13.
    Roffey SJ, Obach RS, Gedge JI, Smith DA. What is the objective of the mass balance study? A retrospective analysis of data in animal and human excretion studies employing radiolabeled drugs. Drug Metab Rev. 2007;39(1):17–43.CrossRefGoogle Scholar
  14. 14.
    Chadrani Gunaratna Bioanalytical Syatems Inc. Drug metabolism and pharmacokinetics in drug discovery: a primer for bioanalytical chemists, part 1. Curr Sep. 2000;19(1):17–23.Google Scholar
  15. 15.
    Bandu R, Ahn HS, Lee JW, Kim YW, Choi SH, Kim HJ, et al. Liquid chromatography electrospray ionization tandem mass spectrometric (LC/ESI–MS/MS) study for the identification and characterization of in vivo metabolites of cisplatin in rat kidney cancer tissues: online hydrogen/deuterium (H/D) exchange study. PLoS One. 2015;10(8):e0134027.CrossRefGoogle Scholar
  16. 16.
    Jeon JS, Oh SJ, Lee JY, Ryu CS, Kim YM, Lee BH, et al. Metabolic characterization of meso-dihydroguaiaretic acid in liver microsomes and in mice. Food Chem Toxicol. 2015;76:94–102.CrossRefGoogle Scholar
  17. 17.
    Peng Y, Wu H, Zhang X, Zhang F, Qi H, Zhong Y, et al. A comprehensive assay for nine major cytochrome P450 enzymes activities with 16 probe reactions on human liver microsomes by a single LC/MS/MS run to support reliable in vitro inhibitory drug–drug interaction evaluation. Xenobiotica. 2015;45(11):961–77.CrossRefGoogle Scholar
  18. 18.
    Sim J, Choi E, Lee YM, Jeong GS, Lee S. In vitro inhibition of human cytochrome P450 by cudratricusxanthone A. Food Chem Toxicol. 2015;81:171–5.CrossRefGoogle Scholar
  19. 19.
    Sun YW, El-Bayoumy K, Aliaga C, Awad AS, Gowda K, Amin S, et al. Tissue distribution, excretion and pharmacokinetics of the environmental pollutant dibenzo[def, p]chrysene in mice. Chem Res Toxicol. 2015;28(7):1427–33.CrossRefGoogle Scholar
  20. 20.
    Bi HC. The pharmacokinetic study of tanshinone IIA in rats and the involved mechanisms. Guangzhou: Sun Yat-sen University; 2007.Google Scholar
  21. 21.
    Erve JC, Vashishtha SC, Ojewoye O, Adedoyin A, Espina R, Demaio W, et al. Metabolism of prazosin in rat and characterization of metabolites in plasma, urine, faeces, brain and bile using liquid chromatography/mass spectrometry (LC/MS). Xenobiotica. 2008;38(5):540–58.CrossRefGoogle Scholar
  22. 22.
    Chen JQ, Wang H, Li DY, Qu XT, Zhu FX. Analysis of metabolites of protocatechuic acid in rats in vivo. Chin Trad Patent Med. 2017;39(03):561–4.Google Scholar
  23. 23.
    Spaggiari D, Geiser L, Daali Y, Rudaz S. A cocktail approach for assessing the in vitro activity of human cytochrome P450s: an overview of current methodologies. J Pharm Biomed Anal. 2014;101:221–37.CrossRefGoogle Scholar
  24. 24.
    Zhong DF, Zhu MS, Humphreys WG. Drug metabolism in drug design and development-basic concepts and practice, vol. 1. Beijing: People’s Military Medical Press; 2011.Google Scholar
  25. 25.
    Lee KS, Kim SK. Direct and metabolism-dependent cytochrome P450 inhibition assays for evaluating drug–drug interactions. J Appl Toxicol. 2013;33(2):100–8.CrossRefGoogle Scholar
  26. 26.
    Jiang YB, Shan JH, Wang Y, Cai YH, Tian YP. Application progress of LC–MS/MS technology in drug metabolism research. Chin J Pharm Anal. 2014;03:385–91.Google Scholar
  27. 27.
    Zhu X, Chen Y, Subramanian R. Comparison of information-dependent acquisition, SWATH, and MS(All) techniques in metabolite identification study employing ultrahigh-performance liquid chromatography–quadrupole time-of-flight mass spectrometry. Anal Chem. 2014;86(2):1202–9.CrossRefGoogle Scholar
  28. 28.
    Xue XY, Lin LF, Xiao F, Pi T, Lai YC, Luo HM. Neurotrophic effects of protocatechuic acid on neurite outgrowth and survival in cultured cerebral cortical neurons of newborn rat. J Chin Med Mater. 2011;34(4):567–71.Google Scholar
  29. 29.
    Park CS, Lee JY, Choi HY, Ju BG, Youn I, Yune TY. Protocatechuic acid improves functional recovery after spinal cord injury by attenuating blood-spinal cord barrier disruption and hemorrhage in rats. Neurochem Int. 2019;124:181–92.CrossRefGoogle Scholar
  30. 30.
    Diao XP, Liao M, Cheng XY, Liang CJ, Sun YP, Zhang X, et al. Identification of metabolites of helicid in vivo using ultra-high performance liquid chromatography–quadrupole time-of-flight mass spectrometry. Biomed Chromatogr. 2018;32:e4263.CrossRefGoogle Scholar
  31. 31.
    Tong JC, Zhou ZM, Qi WW, Jiang SY, Yang B, Zhong ZL, et al. Antidepressant effect of helicid in chronic unpredictable mild stress model in rats. Int Immunopharmacol. 2019;67:13–21.CrossRefGoogle Scholar
  32. 32.
    Maheshwari N, Khan FH, Mahmood R. 3,4-Dihydroxybenzaldehyde lowers ROS generation and protects human red blood cells from arsenic(III) induced oxidative damage. Environ Toxicol. 2018;33:861–73.CrossRefGoogle Scholar
  33. 33.
    Schlickmann F, de Souza P, Boeing T, Mariano LNB, Steimbach VMB, Krueger CMA, et al. Chemical composition and diuretic, natriuretic and kaliuretic effects of extracts of Mimosa bimucronata (DC.) Kuntze leaves and its majority constituent methyl gallate in rats. J Pharm Pharmacol. 2017;69(11):1615–24.CrossRefGoogle Scholar
  34. 34.
    Husain N, Mahmood R. 3,4-Dihydroxybenzaldehyde quenches ROS and RNS and protects human blood cells from Cr(VI)-induced cytotoxicity and genotoxicity. Toxicol In Vitro. 2018;50:293–304.CrossRefGoogle Scholar
  35. 35.
    Bag PK, Roy N, Acharyya S, Saha DR, Koley H, Sarkar P, et al. In vivo fluid accumulation-inhibitory, anticolonization and anti-inflammatory and in vitro biofilm-inhibitory activities of methyl gallate isolated from Terminalia chebula against fluoroquinolones resistant Vibrio cholerae. Microb Pathog. 2019;128:41–6.CrossRefGoogle Scholar
  36. 36.
    Baek JM, Kim JY, Lee CH, Yoon KH, Lee MS. Methyl gallate inhibits osteoclast formation and function by suppressing Akt and Btk-PLCgamma2-Ca (2+) signaling and prevents lipopolysaccharide-induced bone loss. Int J Mol Sci. 2017;18(3):581.CrossRefGoogle Scholar
  37. 37.
    Tai A, Sawano T, Ito H. Antioxidative properties of vanillic acid esters in multiple antioxidant assays. Biosci Biotechnol Biochem. 2012;76:314–8.CrossRefGoogle Scholar
  38. 38.
    Li G, Min BS, Zheng C, Lee J, Oh SR, Ahn KS, et al. Neuroprotective and free radical scavenging activities of phenolic compounds from Hovenia dulcis. Arch Pharm Res. 2005;28(7):804–9.CrossRefGoogle Scholar
  39. 39.
    Cha PH, Shin W, Zahoor M, Kim HY, do Min S, Choi KY. Hovenia dulcis Thunb extract and its ingredient methyl vanillate activate Wnt/beta-catenin pathway and increase bone mass in growing or ovariectomized mice. PloS One. 2014;9(1):e85546.CrossRefGoogle Scholar
  40. 40.
    Lim SJ, Kim M, Randy A, Nam EJ, Nho CW. Effects of Hovenia dulcis Thunb. extract and methyl vanillate on atopic dermatitis-like skin lesions and TNF-alpha/IFN-gamma-induced chemokines production in HaCaT cells. J Pharm Pharmacol. 2016;68(11):1465–79.CrossRefGoogle Scholar
  41. 41.
    Hrycay EG, Bandiera SM. Expression, function and regulation of mouse cytochrome P450 enzymes: comparison with human P450 enzymes. Curr Drug Metab. 2009;10(10):1151–83.CrossRefGoogle Scholar
  42. 42.
    Eagling VA, Tjia JF, Back DJ. Differential selectivity of cytochrome P450 inhibitors against probe substrates in human and rat liver microsomes. Br J Clin Pharmacol. 1998;45(2):107–14.CrossRefGoogle Scholar
  43. 43.
    Zhang PY, Ren J, Zhi WQ, Liu CM, Gao N. Inhibition of CYP450 activity by honokiol and other four components of Chinese traditional medicine in vitro. Chin J Clin Pharmacol Ther. 2017;22(8):922–6.Google Scholar

Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Department of Pharmacology, College of Basic MedicineJinan UniversityGuangzhouChina
  2. 2.State Key Laboratory of Oncology in South ChinaSun Yat-sen University Cancer CenterGuangzhouChina
  3. 3.Analytical and Testing CenterJinan UniversityGuangzhouChina
  4. 4.State Key Laboratory of Applied Microbiology Southern ChinaGuangdong Institute of MicrobiologyGuangzhouChina
  5. 5.SCIEX (China) Co., Ltd.GuangzhouChina
  6. 6.Institute of Brain SciencesJinan UniversityGuangzhouChina

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