Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2022 Jun 8:13:938813.
doi: 10.3389/fphar.2022.938813. eCollection 2022.

JBP485, A Dual Inhibitor of Organic Anion Transporters (OATs) and Renal Dehydropeptidase-I (DHP-I), Protects Against Imipenem-Induced Nephrotoxicity

Affiliations

JBP485, A Dual Inhibitor of Organic Anion Transporters (OATs) and Renal Dehydropeptidase-I (DHP-I), Protects Against Imipenem-Induced Nephrotoxicity

Chong Wang et al. Front Pharmacol. .

Abstract

Imipenem (IMP) possesses a broad spectrum of antibacterial activity; however, nephrotoxicity limits its clinical application in patients with renal insufficiency. In our previous studies, a dipeptide, JBP485, a dipeptide with the chemical structure cyclo-trans-4-L-hydroxyprolyl-L-serine, was found to attenuate drug-induced kidney injury. The current study aimed to explore whether JBP485 could relieve IMP-induced kidney injury and clarify the potential molecular pharmacokinetic mechanism. The effects of JBP485 on IMP nephrotoxicity were evaluated in rabbits and human kidney 2 (HK-2) cells. Drug-drug interactions (DDIs) mediated by organic anion transporters (OATs) and dehydropeptidase-I (DHP-I) were explored through pharmacokinetic studies in rats, metabolism assays in the kidney, and uptake studies in OAT-over-expressing cells. The results revealed that JBP485 significantly ameliorated IMP-induced nephrotoxicity in rabbits. Further, incubation of HK-2 cells with JBP485 or cilastatin markedly improved the cell survival rate, inhibited apoptosis and attenuated mitochondrial damage by improving the stability of IMP and reducing its intracellular accumulation. This suggests that DHP-I and OATs might be involved in the protective effect of JBP485. Furthermore, coadministration with JBP485 significantly increased the IMP's plasma concentration as well as the area under the plasma concentration-time curve (AUC), while decreasing IMP renal clearance and cumulative urinary excretion. Moreover, JBP485 reduced IMP uptake in kidney slices and OAT1/3-human embryonic kidney 293 (HEK293) cells. At the same time, the metabolism of IMP by DHP-I was inhibited by JBP485 with an IC50 value of 12.15 ± 1.22 μM. Finally, the molecular docking assay revealed a direct interaction between JBP485 and OAT1/3 or DHP-I. In conclusion, JBP485 protected against IMP nephrotoxicity in rabbits and HK-2 cells by improving IMP stability and reducing its intracellular accumulation via simultaneous inhibition of renal OATs and DHP-I. JBP485 is a promising renoprotective agent and could serve as an effective supplement to reduce IMP-induced adverse renal reactions in the clinical setting.

Keywords: DDI; DHP-I; Imipenem; JBP485; OATs.

PubMed Disclaimer

Conflict of interest statement

Author TK is employed by Japan Bioproducts Industry Co. Ltd. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

FIGURE 1
FIGURE 1
Protective effect of JBP485 on IMP nephrotoxicity in rabbits. Rabbits were injected with IMP (200 mg/kg) through the ear vein in the absence or presence JBP485 (200 mg/kg). The kidneys were collected 72 h after administration of IMP for gross histological observation (A) and HE staining (B). Blood samples (0.5 ml) were collected for the determination of BUN (C) and CRE (D). Data are expressed as the mean ± SD. **p < 0.01 compared with control; ## p < 0.01 compared with IMP group (n = 3).
FIGURE 2
FIGURE 2
Effect of JBP485 and cilastatin on the cytotoxicity, stability and intracellular accumulation of IMP in HK-2 cells. Protein expression levels of OAT1, OAT3 and DHP-I in HK-2 cells and the mouse kidney by Western blotting (A). HK-2 cells were incubated with IMP (0-5 mM) in the absence or presence of JBP485 (500 μM) or cilastatin (500 μM) for 24 h and cell survival was determined by a CCK-8 assay (B). HK-2 cells were incubated with IMP (2 mM) in the absence or presence of JBP485 (500 μM) or cilastatin (500 μM) for 24 h. Cell apoptosis and the mitochondrial membrane potential were evaluated by Annexin V/PI staining and JC-1 staining assays, respectively (C,D). HK-2 cells were incubated with IMP (50 μM) in the absence or presence of JBP485 (50 μM) or cilastatin (50 μM). The residual concentration of IMP in the medium was determined by LC-MS/MS (E). Intracellular accumulation of IMP in HK-2 cells was determined by LC-MS/MS after incubation with IMP (50 μM) with or without JBP485 (50 μM) or cilastatin (50 μM) for 10 min (F). Data are expressed as the mean ± SD.*p < 0.05 compared with control or IMP group (n = 3).
FIGURE 3
FIGURE 3
Effect of JBP485 on the pharmacokinetics of IMP in rats. Mean plasma concentration-time curves (A), plasma clearance (CLP) (B), cumulative urine excretion curves (C), and renal clearances (CLR) (D) of IMP after intravenous administration of IMP and JBP485 in rats. Data are expressed as the mean ± SD. *p < 0.05 and **p < 0.01 compared with control (n = 5).
FIGURE 4
FIGURE 4
Effect of JBP485 on the uptake of IMP by rat kidney slices and hOAT1/3-HEK293 cells. Inhibition effect of JBP485 (50 μM) on the uptake of IMP (50 μM) in kidney slices (A). Inhibition effect of JBP485 (50 μM) on the uptake of IMP (50 μM) in hOAT1/3-HEK293 cells (B,E). Inhibition effect of JBP485 (50 μM) on the uptake of IMP (10–1,000 μM) in hOAT1/3-HEK293 cells (C,F). Inhibition effect of JBP485 (1–400 μM) on the uptake of IMP (50 μM) in hOAT1/3-HEK293 cells (D,G). Data are expressed as the mean ± SD. *p < 0.05 and ** p < 0.01 compared with control (n = 3).
FIGURE 5
FIGURE 5
Effects of JBP485 on IMP metabolism in rat kidney. Metabolism of IMP (50 μM) by rat kidney in the presence or absence of JBP485 (50 μM) at 0.5, 1, 2 and 3 h (A). Concentration-dependant metabolism of IMP (5–200 μM) with or without JBP485 (50 μM) (B). Inhibitory effect of JBP485 (1–100 μM) on IMP (50 μM) metabolism in rat kidney (C). Data are expressed as the mean ± SD. * p < 0.05 and ** p < 0.01 compared with control (n = 3).
FIGURE 6
FIGURE 6
Molecular docking simulation between JBP485 and DHP-I (A), OAT1 (B) or OAT3 (C).

References

    1. Agudelo M., Rodriguez C. A., Pelaez C. A., Vesga O. (2014). Even Apparently Insignificant Chemical Deviations Among Bioequivalent Generic Antibiotics Can Lead to Therapeutic Nonequivalence: The Case of Meropenem. Antimicrob. Agents Chemother. 58 (2), 1005–1018. 10.1128/AAC.00350-13 - DOI - PMC - PubMed
    1. Bejoy J., Qian E. S., Woodard L. E. (2022). Tissue Culture Models of AKI: From Tubule Cells to Human Kidney Organoids. J. Am. Soc. Nephrol. 33 (3), 487–501. 10.1681/ASN.2021050693 - DOI - PMC - PubMed
    1. Benfield P., Chrisp P. (1992). Imipenem/cilastatin: A Pharmacoeconomic Appraisal of its Use in Intra-abdominal Infections. Pharmacoeconomics 1 (6), 443–459. 10.2165/00019053-199201060-00005 - DOI - PubMed
    1. Birnbaum J., Kahan F. M., Kropp H., MacDonald J. S. (1985). Carbapenems, a New Class of Beta-Lactam Antibiotics. Discovery and Development of Imipenem/cilastatin. Am. J. Med. 78 (6a), 3–21. 10.1016/0002-9343(85)90097-x - DOI - PubMed
    1. Bush K. T., Singh P., Nigam S. K. (2020). Gut-derived Uremic Toxin Handling In Vivo Requires OAT-Mediated Tubular Secretion in Chronic Kidney Disease. JCI Insight 5 (7), e133817. 10.1172/jci.insight.133817 - DOI - PMC - PubMed

LinkOut - more resources