Synthesis of achatin-I (Gly-D-Phe-L-Ala-L-Asp) analogs having dehydrophenylalanine or aminoisobutyric acid residue at position 2, and their effects on Achatina giant neurons.

1. Achatin-I (Gly-D-Phe-L-Ala-L-Asp), a neuroactive tetrapeptide having a D-phenylalanine residue, has been proposed to be an excitatory neurotransmitter of Achatina giant neurons. It was revealed that the D-Phe2 residue is essential for bioactivity of achatin-I, which seems to adopt beta-turn conformation. In the present study, in order to investigate the structure-activity relationships of achatin-I and its derivatives, the two highly constrained analogs of achatin-I, [delta ZPhe2]achatin-I (Gly-delta ZPhe-L-Ala-L-Asp) (delta ZPhe: (Z)-alpha,beta-dehydrophenylalanine) and [Aib2]achatin-I (Gly-Aib-L-Ala-L-Asp) (Aib: alpha-aminoisobutyric acid), were synthesized, and their effects on the two identifiable Achatina giant neuron types, PON (periodically oscillating neuron) and v-RCDN (ventral-right cerebral distinct neuron), were examined in comparison with those of achatin-I under voltage clamp. 2. Achatin-I (n = 6), ejected onto the neurone by brief pneumatic pressure (2 kg/cm2, 400 ms, 10(-3) M, at 10-min intervals), produced an inward current (Im) on PON. The Iin value (mean +/- SEM) was 0.44 +/- 0.03 nA. The interval between the achatin-I ejection and the Iin peak was 14.74 +/- 3.15 s (n = 6). [delta ZPhe2]achatin-I (n = 6) and [Aib2]achatin-I (n = 6) had no effect on this neuron type. 3. On the other hand, achatin-I (n = 10) and [delta ZPhe2]-achatin-I (n = 10), ejected by brief pressure, produced an Iin on v-RCDN. The Iin values were 0.85 +/- 0.07 nA for achatin-I and 0.48 +/- 0.05 nA (p < 0.01, compared with that of achatin-I by Student's t-test for paired data) for [delta ZPhe2]achatin-I. The intervals between the compound ejection and the Iin peak were 5.95 +/- 0.33 s for achatin-I and 8.70 +/- 0.81 s (p < 0.05, compared with that of achatin-I) for [delta ZPhe2]achatin-I. [Aib2]achatin-I (n = 10) had no effect on this neuron type.