Comparison of tachykinin NK1 and NK2 receptors in the circular muscle of the guinea‐pig ileum and proximal colon
Authors: Maggi Carlo Alberto , Patacchini Riccardo , Meini Stefania , Quartara Laura , Sisto Alessandro , Potier Edoardo , Giuliani Sandro , Giachetti Antonio 
Published: July 2012

The aim of this study was the pharmacological characterization of tachykinin NK1 and NK2 receptors mediating contraction in the circular muscle of the guinea‐pig ileum and proximal colon. The action of substance P (SP), neurokinin A (NKA) and of the synthetic agonists [Sar9]SP sulphone, [Glp6,Pro9]SP(6–11) (septide) and [βAla8]NKA(4–10) was investigated. The affinities of various peptide and nonpeptide antagonists for the NK1 and NK2 receptor was estimated by use of receptor selective agonists.

The natural agonists, SP and NKA, produced concentration‐dependent contraction in both preparations. EC50 values were 100 pm and 5 nm for SP, 1.2 nm and 19 nm for NKA in the ileum and colon, respectively. The action of SP and NKA was not significantly modified by peptidase inhibitors (bestatin, captopril and thiorphan, 1 μm each).

Synthetic NK1 and NK2 receptor agonists produced concentration‐dependent contraction of the circular muscle of the ileum and proximal colon. EC50 values were 83 pm, 36 pm and 10 nm in the ileum, 8 nm, 0.7 nm and 12 nm in the colon for [Sar9]SP sulphone, septide and [βAla8]NKA(4–10), respectively. The pseudopeptide derivative of NKA(4–10), MDL 28,564 behaved as a full or near‐to‐full agonist in both preparations, its EC50s being 474 nm and 55 nm in the ileum and colon, respectively.

Nifedipine (1 μm) abolished the response to septide and [Sar9]SP sulphone in the ileum and produced a rightward shift and large depression of the response in the colon. The response to [βAla8]NKA(4–10) was abolished in the ileum and largely unaffected in the colon.

The NK1 receptor antagonists, (±)‐CP 96,34, FK 888 and GR 82,334 competitively antagonized the response to septide and [Sar9]SP sulphone in both preparations without affecting that to [βAla8]NKA(4–10). In general, the NK1 receptor antagonists were significantly more potent toward septide than [Sar9]SP sulphone in both preparations.

The NK2 receptor antagonists, GR 94,800 and SR 48,968 selectively antagonized the response to [βAla8]NKA(4–10) without affecting that to [Sar9]SP sulphone or septide in the ileum and colon. SR 48,968 produced noncompetitive antagonism of the response to the NK2 receptor agonist in the ileum and competitive antagonism in the colon.

MEN 10,376 and the cyclic pseudopeptide MEN 10,573 antagonized in a competitive manner the response to [βAla8]NKA(4–10) in the ileum and colon. While MEN 10,573 was equipotent in both preparations, MEN 10,376 was significantly more potent in the colon than in the ileum. MEN 10,376 was also effective against septide in both preparations, without affecting the response to [Sar9]SP sulphone. MEN 10,573 antagonized the response to [Sar9]SP sulphone and septide in both preparations, pKB values against septide being intermediate, and significantly different from, those measured against [βAla8]NKA(4–10) and [Sar9]SP sulphone.

These findings show that tachykinin NK1 and NK2 receptors mediate contraction of the circular muscle of the guinea‐pig ileum and colon. In both preparations NK1 receptor antagonists display higher apparent affinity when tested against septide than [Sar9]SP sulphone. These findings are compatible with the proposed existence of NK1 receptor subtypes in guinea‐pig, although alternative explanations (e.g. agonist binding to different epitopes of the same receptor protein) cannot be excluded at present. Furthermore, an intraspecies heterogeneity of the NK2 receptor in the circular muscle of the guinea‐pig ileum and colon is suggested.