1. Synthesis and biological evaluation of selected insect neuropeptide analogs modified by D- or L-phenylglycine derivatives
W Szeszel-Fedorowicz, G Rosiński, J Issberner, R Osborne, I Janssen, A De Loof, D Konopińska Acta Pol Pharm. 2000 Nov;57 Suppl:88-9.
Novel analogs, modified by L- or D- phenylglycine and p-substituted derivatives, of the neuromodulator proctolin (Arg-Tyr-Leu-Pro-Thr) and of the Trypsin Modulating Oostatic Factor from the gray flesh fly Neobellieria bullata (Neb-TMOF-Asn-Pro-Thr-Asn-Leu-His) were synthesized and checked for activity. Proctolin analogs were modified at position 2: Arg-Phg-Leu-Pro-Thr (I), Arg-D-Phg-Leu-Pro-Thr (II), Arg-Phg(p-OH)-Leu-Pro-Thr (III), Arg-D-Phg(p-OH)-Leu-Pro-Thr (IV), Arg-Phg(p-NO2)-Leu-Pro-Thr (V) Arg-D-Phg(p-NO2)-Leu-Pro-Thr (VI), Arg-Phg(p-NH2)-Leu-Pro-Thr (VII), Arg-D-Phg(p-NH2)-Leu-Pro-Thr (VIII), Arg-Phg(p-N,N-di-Me)-Leu-Pro-Thr (IX), Arg-D-Phg(pp-N,N-di-Me)-Leu-Pro-Thr (X) while analogs of Neb-TMOF underwent modifications at position 6: Asn-Pro-Thr-Asn-Leu-Phg(p-NO2) (XI), Asn-Pro-Thr-Asn-Leu-D-Phg(p-NO2) (XII), Asn-Pro-Thr-Asn-Leu-Phg(p-NH2) (XIII), Asn-Pro-Thr-Asn-Leu-D-Phg(p-NH2) (XIV), Asn-Pro-Thr-Asn-Leu-Phg(p-N,N-di-Me) (XV), Asn-Pro-Thr-Asn-Leu-D-Phg(p-N,N-di-Me) (XVI). Earlier studies on proctolin demonstrated that the presence of the -CH2- group between C-alpha and the phenyl ring at position 2 of the peptide chain is important for the myotropic activity. Based on these results, we replaced Tyr at position 2 by different phenylglycine derivatives, lacking the methylene group at the side chain. Myotropic activity of the proctolin analogs was assayed in vitro on the semi-isolated heart of the mealworm Tenebrio molitor and on the foregut of the locust Schistocerca gregaria. All analogs (I-X) were practically inactive. For Neb-TMOF, it was previously demonstrated that the exchange of His-6 by p-substituted Phe-derivatives, especially by Phe(p-NH2), an amino acid containing a basic function, results into analogs which inhibit trypsin biosynthesis in the gray fleshfly. For this reason these new Neb-TMOF analogs with L- or D-phenylglycine p-substituted derivatives at position 6, were developed and tested (in vivo) in the trypsin biosynthesis assay of the gray fleshfly N. bullata. Only analogs XV and XVI slightly inhibited trypsin biosynthesis in the midgut. Because more than 50% of the injected animals died and none of the surviving animals ate much of the liver meal, the lower trypsin level in the gut might be a indirect effect. Other peptides (XI-XIV) had no effect on the level of trypsin biosynthesis in the midgut.
2. Active fragments and analogs of the plant growth factor, phytosulfokine: structure-activity relationships
Y Matsubayashi, H Hanai, O Hara, Y Sakagami Biochem Biophys Res Commun. 1996 Aug 5;225(1):209-14. doi: 10.1006/bbrc.1996.1155.
Sulfated pentapeptide phytosulfokine-alpha (PSK-alpha) is the first chemically characterized peptidal plant growth factor that induces proliferation of the mesophyll cells of asparagus. To determine the active core involved in the sequence PSK-alpha, we synthesized PSK-alpha and twelve PSK-alpha analogs by the solid phase peptide synthesis and the direct sulfation to the peptide-resin using dimethylformamide-sulfurtrioxide (DMF-SO3) complex. The truncated analogs of PSK-alpha without the first and second C-terminal amino acids retained 8% and 20% of the activity of the parent pentapeptide, respectively. Deletion of the sulfate groups of Tyr1 and Tyr3 resulted in compounds with 0.6% and 4% of the activity of PSK-alpha, indicating that the sulfate group of Tyr1 is more important than that of Tyr3 for the expression of its activity. In contrast, the N-terminal truncated analog and unsulfated analog exhibited little or no activity. Thus the N-terminal tripeptide fragment H-Tyr(SO3H)-Ile-Tyr(SO3H)-OH has been identified as the active core of PSK-alpha.
3. Further analogues of plant peptide hormone phytosulfokine-alpha (PSK-alpha) and their biological evaluation
Agata Bahyrycz, Yoshikatsu Matsubayashi, Mari Ogawa, Youji Sakagami, Danuta Konopińska J Pept Sci. 2005 Sep;11(9):589-92. doi: 10.1002/psc.653.
Phytosulfokine-alpha (PSK-alpha), a sulfated growth factor of structure H-Tyr(SO3H)-Ile-Tyr(SO3H)-Thr-Gln-OH universally found in both monocotyledons and dicotyledons, strongly promotes proliferation of plant cells in culture. In studies on the structure/activity relationship of PSK-alpha the synthesis was performed of a series of a further 23 analogues modified in position 1, 3 or 4 as well as simultaneously in positions 1 and 3 of the peptide chain. Peptides were synthesized by the solid phase method according to the Fmoc procedure on a Wang-resin. Free peptides were released from the resin by 95% TFA in the presence of EDT. All peptides were tested by competitive binding assay to the carrot membrane using 3H-labelled PSK-alpha according to the test of Matsubayashi et al. Among these peptide analogues, [H-Phe(4-Cl)1]-PSK-alpha (IV), [H-Phe(4-I)1]-PSK-alpha (VII), and [Phe(4-Cl)3]-PSK-alpha (XI) retained 30% PSK-alpha activity. Analogue [Tyr(PO3H2)3]-PSK-alpha (IX) showed 10% of PSK-alpha activity.
