Interaction of N-benzoyl-D-phenylalanine and related compounds with the sulphonylurea receptor of beta-cells

Br J Pharmacol. 1998 Mar;123(6):1023-30. doi: 10.1038/sj.bjp.0701686.

Abstract

1. The structure activity relationships for the insulin secretagogues N-benzoyl-D-phenylalanine (NBDP) and related compounds were examined at the sulphonylurea receptor level by use of cultured HIT-T15 and mouse pancreatic beta-cells. The affinities of these compounds for the sulphonylurea receptor were compared with their potencies for K(ATP)-channel inhibition. In addition, the effects of cytosolic nucleotides on K(ATP)-channel inhibition by NBDP were investigated. 2. NBDP displayed a dissociation constant for binding to the sulphonylurea receptor (K(D) value) of 11 microM and half-maximally effective concentrations of K(ATP)-channel inhibition (EC50 values) between 2 and 4 microM (in the absence of cytosolic nucleotides or presence of 0.1 mM GDP or 1 mM ADP). 3. In the absence of cytosolic nucleotides or presence of GDP (0.1 mM) maximally effective concentrations of NBDP (0.1-1 mM) reduced K(ATP)-channel activity to 47% and 44% of control, respectively. In the presence of ADP (1 mM), K(ATP)-channel activity was completely suppressed by 0.1 mM NBDP. 4. The L-isomer of N-benzoyl-phenylalanine displayed a 20 fold lower affinity and an 80 fold lower potency than the D-isomer. 5. Introduction of a p-nitro substituent in the D-phenylalanine moiety of NBDP did not decrease lipophilicity but lowered affinity and potency by more than 30 fold. 6. Introduction of a p-amino substituent in the D-phenylalanine moiety of NBDP (N-benzoyl-p-amino-D-phenylalanine, NBADP) reduced lipophilicity and lowered affinity and potency by about 10 fold. This loss of affinity and potency was compensated for by formation of the phenylpropionic acid derivative of NBADP. A similar difference in affinity was observed for the sulphonylurea carbutamide and its phenylpropionic acid derivative. 7. Replacing the benzene ring in the D-phenylalanine moiety of NBDP by a cyclohexyl ring increased lipophilicity, and the K(D) and EC50 values were slightly lower than for NBDP. Exchange of both benzene rings in NBDP by cyclohexyl rings further increased lipophilicity without altering affinity and potency. 8. This study shows that N-acylphenylalanines interact with the sulphonylurea receptor of pancreatic beta-cells in a stereospecific manner. Their potency depends on lipophilic but not aromatic properties of their benzene rings. As observed for sulphonylureas, interaction of N-acylphenylalanines with the sulphonylurea receptor does not induce complete inhibition of K(ATP)-channel activity in the absence of inhibitory cytosolic nucleotides.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP-Binding Cassette Transporters*
  • Adenosine Triphosphate / metabolism
  • Animals
  • Calcium / metabolism
  • Carbutamide / analogs & derivatives
  • Carbutamide / pharmacology
  • Cell Line, Transformed
  • Cricetinae
  • Cyclohexanes / pharmacology
  • Glyburide / metabolism
  • Guanosine Diphosphate / metabolism
  • Islets of Langerhans / cytology
  • Islets of Langerhans / drug effects
  • Islets of Langerhans / metabolism*
  • Male
  • Mice
  • Nateglinide
  • Phenylalanine / analogs & derivatives*
  • Phenylalanine / metabolism
  • Phenylalanine / pharmacology
  • Potassium Channels / drug effects
  • Potassium Channels / metabolism*
  • Potassium Channels, Inwardly Rectifying*
  • Receptors, Drug / metabolism*
  • Sulfonylurea Receptors
  • Tritium

Substances

  • ATP-Binding Cassette Transporters
  • Cyclohexanes
  • Kcnj1 protein, mouse
  • Potassium Channels
  • Potassium Channels, Inwardly Rectifying
  • Receptors, Drug
  • Sulfonylurea Receptors
  • Tritium
  • Guanosine Diphosphate
  • Nateglinide
  • Phenylalanine
  • Adenosine Triphosphate
  • benzoylphenylalanine
  • Carbutamide
  • Glyburide
  • Calcium