Antisense candidates against protein kinase C-alpha designed based on phylogenesis and simulant structure of mRNA.

Abstract

AIM:

To optimize the antisense drug design by the combined method of phylogenetic analysis and secondary structure prediction and to get ideal candidates.

METHODS:

The phylogenetic analysis and the secondary structure simulation were performed by computer. Oligodeoxynucleotides (ODN) were designed against the full-conserved blocks with low local reaction free energy of protein kinase C (PKC)-alpha mRNA. The in vitro effects of ODN were evaluated by human A549 lung carcinoma cells and mouse B16-BL6 melanoma cells, the expression of target mRNA was detected by in situ hybridization and RT-PCR. The in vivo effects of ODN were also evaluated by models of A549 xenografts in nude mice and B16 melanoma in mice.

RESULTS:

Three ODN had significantly lower IC50 values than that of ISIS3521, the positive control, on A549 cells in vitro. Five ODN inhibited the growth of B16-BL6 cells with IC50 <100 nmol/L, while IC50 of ISIS3521 was >200 nmol/L. In situ hybridization and RT-PCR showed that the best candidate AP1261 inhibited the expression of PKC-alpha at mRNA level in a dose-dependent manner. AP1261 inhibited the growth of A549 and B16 tumors in vivo at 0.005-0.5 mg.kg(-1).d(-1). The inhibitory rate of AP1261 on A549 tumors was greater than that of ISIS3521 at the same dose. ISIS3521 did not affect the growth of B16 tumors.

CONCLUSION:

AP1261 may be of value as an antitumor agent or adjuvant and the combined method of phylogenetic analysis and secondary structure prediction is a potential helpful tool for antisense drug design.

• Authors: Hai-Feng Song, Zhong-Ming Tang, Shou-Jun Yuan, Bao-Zhen Zhu, Xiu-Wen Liu
• Journal: Acta pharmacologica Sinica (Acta Pharmacol Sin) Vol. 24 Issue 3 Pg. 269-76 (Mar 2003) ISSN: 1671-4083 [Print] United States
• PMID: 12617778 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antineoplastic Agents
• Isoenzymes
• Oligodeoxyribonucleotides, Antisense
• RNA, Messenger
• PRKCA protein, human
• Prkca protein, mouse
• Protein Kinase C
• Protein Kinase C-alpha

Topics

• Adenocarcinoma (metabolism, pathology)
• Animals
• Antineoplastic Agents (chemical synthesis, pharmacology)
• Cell Division (drug effects)
• Cell Line, Tumor
• Computer-Aided Design
• Drug Design
• Humans
• Isoenzymes (chemistry, pharmacology)
• Lung Neoplasms (metabolism, pathology)
• Melanoma, Experimental (pathology)
• Mice
• Mice, Nude
• Neoplasm Transplantation
• Oligodeoxyribonucleotides, Antisense (chemical synthesis, pharmacology)
• Protein Kinase C (biosynthesis, chemistry, genetics)
• Protein Kinase C-alpha
• Protein Structure, Secondary
• RNA, Messenger (biosynthesis, genetics)
• Rats