Quantification of soluble epoxide hydrolase inhibitors in experimental and clinical samples using the nanobody-based ELISA

Huiyi Yang; Meng Qi; Qiyi He; Sung Hee Hwang; Jun Yang; Mark McCoy; Christophe Morisseau; Suqing Zhao; Bruce D. Hammock

doi:10.1016/j.jpha.2023.05.006

Volume 13 Issue 9

Sep. 2023

Turn off MathJax

Article Contents

Article Navigation > Journal of Pharmaceutical Analysis > 2023 > 13(9): 1013-1023

Huiyi Yang, Meng Qi, Qiyi He, Sung Hee Hwang, Jun Yang, Mark McCoy, Christophe Morisseau, Suqing Zhao, Bruce D. Hammock. Quantification of soluble epoxide hydrolase inhibitors in experimental and clinical samples using the nanobody-based ELISA[J]. Journal of Pharmaceutical Analysis, 2023, 13(9): 1013-1023. doi: 10.1016/j.jpha.2023.05.006

Citation:

Huiyi Yang, Meng Qi, Qiyi He, Sung Hee Hwang, Jun Yang, Mark McCoy, Christophe Morisseau, Suqing Zhao, Bruce D. Hammock. Quantification of soluble epoxide hydrolase inhibitors in experimental and clinical samples using the nanobody-based ELISA[J]. Journal of Pharmaceutical Analysis, 2023, 13(9): 1013-1023. doi: 10.1016/j.jpha.2023.05.006

Citation:

Huiyi Yang, Meng Qi, Qiyi He, Sung Hee Hwang, Jun Yang, Mark McCoy, Christophe Morisseau, Suqing Zhao, Bruce D. Hammock. Quantification of soluble epoxide hydrolase inhibitors in experimental and clinical samples using the nanobody-based ELISA[J]. Journal of Pharmaceutical Analysis, 2023, 13(9): 1013-1023. doi: 10.1016/j.jpha.2023.05.006

PDF( 2193 KB)

Quantification of soluble epoxide hydrolase inhibitors in experimental and clinical samples using the nanobody-based ELISA

doi: 10.1016/j.jpha.2023.05.006

a. Department of Pharmaceutical Engineering, School of Biomedical and Pharmaceutical Sciences, Guangdong University of Technology, Guangzhou, China;
b. Department of Entomology and Nematology and UCD Comprehensive Cancer Center, University of California, Davis, CA 95616, USA;
c. Langfang Normal University, Langfang, Hebei, 065000, China

Funds:

This work was supported by NIEHS (RIVER Award, R35 ES030443), NIEHS (Superfund Award, P42 ES004699), NINDS (Counter Act Program U54 NS127758), Juvenile Diabetes Research Foundation (2-SRA-2022-1210-S-B), Guangzhou Science and Technology Foundation (Grant No.: 201903010034), Natural Resources Science Foundation of Guangdong Province (Grant No.: 2018A030313926), Science and Technology Foundation Key R&D Program of Guangdong Province (Grant Nos.: 2019B020209009 and 2019B020218009), R&D Program of Guangdong Province Drug Administration (Grant Nos.: 2021TDZ09 and 2021YDZ06). This work was also supported by China Scholarship Council (CSC) (202108440382).

Received Date: Mar. 16, 2023
Accepted Date: May 12, 2023
Rev Recd Date: May 08, 2023
Publish Date: May 16, 2023

Abstract

Abstract

To ensure proper dosage of a drug, analytical quantification of it in biofluid is necessary. Liquid chromatography mass spectrometry (LC-MS) is the conventional method of choice as it permits accurate identification and quantification. However, it requires expensive instrumentation and is not appropriate for bedside use. Using soluble epoxide hydrolase (sEH) inhibitors (EC5026 and TPPU) as examples, we report development of a nanobody-based enzyme-linked immunosorbent assay (ELISA) for such small molecules and its use to accurately quantify the drug chemicals in human samples. Under optimized conditions, two nanobody-based ELISAs were successfully established for EC5026 and TPPU with low limits of detection of 0.085 ng/mL and 0.31 ng/mL, respectively, and two order of magnitude linear ranges with high precision and accuracy. The assay was designed to detect parent and two biologically active metabolites in the investigation of a new drug candidate EC5026. In addition, the ELISAs displayed excellent correlation with LC-MS analysis and evaluation of inhibitory potency. The results indicate that nanobody-based ELISA methods can efficiently analyze drug like compounds. These methods could be easily implemented by the bedside, in the field in remote areas or in veterinary practice. This work illustrates that nanobody based assays offer alternative and supplementary analytical tools to mass spectrometry for monitoring small molecule medicines during clinical development and therapy. Attributes of nanobody based pharmaceutical assays are discussed.
- Nanobody,
- Immunoassay,
- Soluble epoxide hydrolase inhibitors,
- Metabolites,
- Small molecules

FullText(HTML)

References(35)

References

C. Morisseau, B.D. Hammock, Impact of soluble epoxide hydrolase and epoxyeicosanoids on human health, Annu. Rev. Pharmacol. Toxicol. 53 (2013) 37-58.

C. Morisseau, B.D. Hammock, Epoxide hydrolases: Mechanisms, inhibitor designs, and biological roles, Annu. Rev. Pharmacol. Toxicol. 45 (2005) 311-333.

L. Yao, B. Cao, Q. Cheng, et al., Inhibition of soluble epoxide hydrolase ameliorates hyperhomocysteinemia-induced hepatic steatosis by enhancing beta-oxidation of fatty acid in mice, Am. J. Physiol. Gastrointest Liver Physiol. 316 (2019) G527-G538.

D. Xu, N. Li, Y. He, et al, Prevention and reversal of cardiac hypertrophy by soluble epoxide hydrolase inhibitors, PNAS 103 (2006) 18733-18738.

I. Bastan, X.N. Ge, M. Dileepan, et al., Inhibition of soluble epoxide hydrolase attenuates eosinophil recruitment and food allergen-induced gastrointestinal inflammation, J. Leukoc. Biol. 104 (2018) 109-122.

S.-K. Anandan, H.K. Webb, D. Chen, et al., 1-(1-acetyl-piperidin-4-yl)-3-adamantan-1-yl-urea (AR9281) as a potent, selective, and orally available soluble epoxide hydrolase inhibitor with efficacy in rodent models of hypertension and dysglycemia, Bioorg. Med. Chem. Lett. 21 (2011) 983-988.

B.D Hammock, K.Wagner, B. Inceoglu, The soluble epoxide hydrolase as a pharmaceutical target for pain management, Pain Management 1 (2011) 383-386.

J. Hu, S. Dziumbla, J. Lin, et al., Inhibition of soluble epoxide hydrolase prevents diabetic retinopathy, Nature 552 (2017) 248-252.

S. Zarriello, J.P. Tuazon, S. Corey, et al., Humble beginnings with big goals: Small molecule soluble epoxide hydrolase inhibitors for treating cns disorders, Prog. Neurobiol. 172 (2019) 23-39.

B.D. Hammock, C.B. McReynolds, K. Wagner, et al., Movement to the clinic of soluble epoxide hydrolase inhibitor EC5026 as an analgesic for neuropathic pain and for use as a nonaddictive opioid alternative, J. Med. Chem. 64 (2021) 1856-1872.

T.E. Rose, C. Morisseau, J.-Y. Liu, et al., 1-aryl-3-(1-acylpiperidin-4-yl)urea inhibitors of human and murine soluble epoxide hydrolase: Structure-activity relationships, pharmacokinetics, and reduction of inflammatory pain, J. Med. Chem. 53 (2010) 7067-7075.

D. Wan, J. Yang, C.B. McReynolds, et al., In vitro and in vivo metabolism of a potent inhibitor of soluble epoxide hydrolase, 1-(1-propionylpiperidin-4-yl)-3-(4-(trifluoromethoxy)phenyl)urea, Front. Pharmacol. 10 (2019), 464.

Y. Li, H. Zhang, Z. Cui, et al., Chemical staining enhanced enzyme-linked immunosorbent assay for sensitive determination of clenbuterol in food, Food Chem. 400 (2023), 134012.

P. Peng, C. Liu, Z. Li, et al., Emerging elisa derived technologies for in vitro diagnostics, TrAC Trends in Anal. Chem. 152 (2022), 116605.

K.L. Singampalli, J. Li, P.B. Lillehoj, Rapid magneto-enzyme-linked immunosorbent assay for ultrasensitive protein detection, Anal. Chim. Acta 1225 (2022), 340246.

M. Baker, Blame it on the antibodies, Nature 521 (2015), 274.

C. Hamers-Casterman, T. Atarhouch, S. Muyldermans, et al., Naturally occurring antibodies devoid of light chains, Nature 363 (1993) 446-448.

S. Steeland, R.E. Vandenbroucke, C. Libert, Nanobodies as therapeutics: Big opportunities for small antibodies, Drug Discov. Today 21 (2016) 1076-1113.

B. Li, X. Qin, L.-Z. Mi, Nanobodies: From structure to applications in non-injectable and bispecific biotherapeutic development, Nanoscale 14 (2022) 7110-7122.

Y. Wang, Y. Xianyu, Nanobody and nanozyme-enabled immunoassays with enhanced specificity and sensitivity, Small Methods 6 (2022), 2101576.

E.A. Bastos-Soares, R.M.O. Sousa, A.F. Gomez, et al., Single domain antibodies in the development of immunosensors for diagnostics, Int. J. Biol. Macromol. 165 (2020) 2244-2252.

N. Alvarez-Rueda, G. Behar, V. Ferre, et al., Generation of llama single-domain antibodies against methotrexate, a prototypical hapten, Mol. Immunol. 44 (2007) 1680-1690.

K.S.S. Lee, J.C. Ng, J. Yang, et al., Preparation and evaluation of soluble epoxide hydrolase inhibitors with improved physical properties and potencies for treating diabetic neuropathic pain, Bioorg. Med. Chem. 28 (2020), 115735.

S.H. Hwang, H.J. Tsai, J.Y. Liu, et al., Orally bioavailable potent soluble epoxide hydrolase inhibitors, J. Med. Chem. 50 (2007) 3825-3840.

P.D. Jones, H.-J. Tsai, Z.N. Do, et al., Synthesis and SAR of conformationally restricted inhibitors of soluble epoxide hydrolase, Bioorg. Med. Chem. Lett. 16 (2006) 5212-5216.

I.-H. Kim, H.-J. Tsai, K. Nishi, et al., 1,3-disubstituted ureas functionalized with ether groups are potent inhibitors of the soluble. Epoxide hydrolase with improved pharmacokinetic properties, J. Med. Chem. 50 (2007) 5217-5226.

N. Lee, D.P. McAdam, J.H. Skerritt, Development of immunoassays for type II synthetic pyrethroids. 1. Hapten design and application to heterologous and homologous assays, J. Agric. Food Chem. 46 (1998) 520-534.

D. Zabetakis, M.A. Olson, G.P. Anderson, et al., Evaluation of disulfide bond position to enhance the thermal stability of a highly stable single domain antibody, PLOS One 9 (2014), e115405.

K.B. Turner, D. Zabetakis, E.R. Goldman, et al., Enhanced stabilization of a stable single domain antibody for seb toxin by random mutagenesis and stringent selection, Protein Eng. Des. Sel. 27 (2014) 89-95.

J.M.J. Pérez, J.G. Renisio, J.J. Prompers, et al., Thermal unfolding of a llama antibody fragment a two-state reversible process, Biochemistry 40 (2001) 74-83.

A. Guedes, L. Galuppo, D. Hood, et al., Soluble epoxide hydrolase activity and pharmacologic inhibition in horses with chronic severe laminitis, Equine. Vet. J. 49 (2017) 345-351.

A. Ulu, S.E. Appt, C. Morisseau, et al., Pharmacokinetics and in vivo potency of soluble epoxide hydrolase inhibitors in cynomolgus monkeys, Br. J. Pharmacol. 165 (2012) 1401-1412.

C.P. Sun, X.Y. Zhang, C. Morisseau, et al., Discovery of soluble epoxide hydrolase inhibitors from chemical synthesis and natural products, J. Med. Chem. 64 (2021) 184-215.

N. Singh, D. Li, C.B. McReynolds, et al., Improved elisa for linoleate-derived diols in human plasma utilizing a polyhrp-based secondary tracer, Anal. Methods 14 (2022) 1810-1819.

P.D. Jones, N.M. Wolf, C. Morisseau, et al., Fluorescent substrates for soluble epoxide hydrolase and application to inhibition studies, Anal. Biochem. 343 (2005) 66-75.

Relative Articles

Supplements(0)

Cited By

Proportional views