[Lys5,MeLeu9,Nle10]-NKA(4–10) induces neurokinin 2 receptor mediated urination and defecation and neurokinin 1 receptor mediated flushing in rats: measured using the rapid detection voiding assay

Jason B. Cook; Raymond Piatt; Lesley Marson

doi:10.1515/jbcpp-2022-0119

Article

[Lys⁵,MeLeu⁹,Nle¹⁰]-NKA_(4–10) induces neurokinin 2 receptor mediated urination and defecation and neurokinin 1 receptor mediated flushing in rats: measured using the rapid detection voiding assay

Jason B. Cook , Raymond Piatt and Lesley Marson

Published/Copyright: November 16, 2022

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From the journal Journal of Basic and Clinical Physiology and Pharmacology Volume 34 Issue 2

Abstract

Objectives

Neurokinin 2 receptor (NK2R) agonists may be useful for treating bladder and bowel dysfunction via direct contraction of detrusor and gastrointestinal smooth muscle. The NK2R agonist [Lys5, MeLeu9, Nle10]-NKA(4–10) (LMN-NKA) induces urination and defecation, but also produces the potential side effect of dermal flushing in rats. Although LMN-NKA is a NK2R agonist, it also has affinity for neurokinin 1 receptors (NK1R). Therefore, the goal of this study was to determine the neurokinin receptor (NKR) subtypes responsible for LMN-NKA-induced urination, defecation, and flushing by blocking either NK2Rs or NK1Rs before LMN-NKA administration.

Methods

To accomplish this goal, we developed a simple high-throughput ‘rapid detection voiding assay’ to detect rapid-onset drug-induced urination and defecation in rats. In LMN-NKA dose-response experiments, LMN-NKA (10–100 μg/kg, subcutaneous) was injected and urination, defecation, and flushing were monitored for 30 min. For NKR antagonist experiments, vehicle, the NK2R antagonist GR159897, or the NK1R antagonist CP-99,994 were injected before an acclimation period. Following acclimation, saline or 100 μg/kg LMN-NKA were injected, and behavior was observed for 30 min.

Results

LMN-NKA produced dose-related increases in urination, defecation, and flushing. Blocking NK2Rs reduced urination and blocked defecation, without affecting flushing. Blocking NK1Rs did not change LMN-NKA-induced urination or defecation but reduced LMN-NKA-induced flushing.

Conclusions

Using the rapid detection voiding assay we show that LMN-NKA-induced urination and defecation are mediated by NK2Rs, while flushing is mediated by NK1Rs. Therefore, drugs that are more selective for NK2 vs. NK1Rs should produce rapid-onset urination and defecation without producing the potential side effect of flushing.

Keywords: defecation; neurokinin 1 receptor; neurokinin 2 receptor; rapid detection voiding assay; urination

Corresponding author: Jason B. Cook, PhD, Dignify Therapeutics LLC, 2 Davis Drive, PO Box 13169, Research Triangle Park, NC 27709, USA, Phone: (919) 371 8138, E-mail: jcook@dignifytherapeutics.com

Funding source: National Institute of Diabetes and Digestive and Kidney Diseases

Award Identifier / Grant number: R44DK112437

Acknowledgments

We thank Integrated Laboratory Systems for their collaboration. We also thank Dr. Nadia Rupniak for a thorough and thoughtful review of the manuscript.

Research funding: This study was funded by the NIDDK of the National Institutes of Health under award number R44DK112437.
Author contributions: JBC performed experiments, collected and analyzed the data, wrote the manuscript. RP performed experiments, collected data, edited manuscript. LM conceived the study, edited manuscript, acquired funding.
Competing interests: Jason B. Cook, Raymond Piatt, and Lesley Marson are employed by Dignify Therapeutics LLC. Lesley Marson has equity ownership in Dignify Therapeutics LLC.
Informed consent: Not applicable.
Ethical approval: Experiments conformed with NIH Guidelines for the Care and Use of Laboratory Animals and were approved by the Integrated Laboratory Systems Animal Care and Use Committee (Animal approved project number 2018-03).

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Received: 2022-05-16

Accepted: 2022-10-19

Published Online: 2022-11-16

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Articles in the same Issue

https://doi.org/10.1515/jbcpp-2022-0119

Keywords for this article

defecation; neurokinin 1 receptor; neurokinin 2 receptor; rapid detection voiding assay; urination