Synthesis and assay of isoquinoline derivatives as HIV-1 Tat-TAR interaction inhibitors

Bioorganic and Medicinal Chemistry Letters

Volumn 15, Issue 17, 2005, Pages 3978-3981

  He, Meizi ^a   Yuan, Dekai ^a   Lin, Wei ^a   Pang, Ruifang ^a   Yu, Xiaolin ^a   Yang, Ming ^a  

^a PEKING UNIVERSITY   (China)

Author keywords

CE; Isoquinoline; Tat TAR interaction

Indexed keywords

GUANIDINE; ISOQUINOLINE DERIVATIVE; PROTEIN INHIBITOR; RNA; TRANSACTIVATOR PROTEIN;

ARTICLE; DRUG BINDING; DRUG PROTEIN BINDING; DRUG SYNTHESIS; DRUG TARGETING; HUMAN IMMUNODEFICIENCY VIRUS 1; LONG TERMINAL REPEAT; PROTEIN RNA BINDING; SIMIAN IMMUNODEFICIENCY VIRUS;

ANTI-HIV AGENTS; CELL LINE; ELECTROPHORESIS, CAPILLARY; GENE PRODUCTS, TAT; HIV LONG TERMINAL REPEAT; HUMANS; ISOQUINOLINES; PROTEIN BINDING; STRUCTURE-ACTIVITY RELATIONSHIP;

HUMAN IMMUNODEFICIENCY VIRUS 1; SIMIAN IMMUNODEFICIENCY VIRUS;

EID: 23244444316     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.bmcl.2005.01.068     Document Type: Article

References (18)

1. T.M. Rana, and K. Jeang Arch. Biochem. Biophys. 365 1999 175
2. J.A. Turpin Expert Rev. Anti-infect. 1 2003 97
3. F. Aboul-ela, J. Karn, and G. Varani Nucleic Acid Res. 24 1996 3974
4. B.J. Calnan, B. Tidor, S. Biancalana, D. Hudson, and A.D. Frankel Science 252 1991 1167
5. J. Karn, and M.A. Graeble Trends Genet. 8 1992 365
6. N. Tassew, and M. Thompson Anal. Chem. 74 2002 5313
7. C.S. Chow, and F. Bogdan Chem. Rev. 97 1997 1489
8. Kato, H.; Miyazawa, K.; Etchu, E. Japan. 7307, 115 (C1. C07d, A61k).
9. M. Cain, R.W. Weber, and F.J. Guzman Med. Chem. 25 1982 1081
10. 2: (FABMS) (m/z): 271.9 (M+H); mp: 136-137°C.
11. B. Zhang, H. Xue, and B.C. Lin Chromatographia 48 1998 268
12. P. Mucha, A. Szyk, P. Rekowski, and J. Barciszewski J. Chromatogr. A 968 2002 211
13. H. Zhao, D. Dai, J. Li, Y. Chen, and L. Jiang BBRC 312 2003 351
14. X. Yu, W. Lin, J. Lin, and M. Yang Bioorg. Med. Chem. Lett. 14 2004 3127
15. Capillary electrophoresis assay: CE experiments were carried out on a Beckman P/ACE 2100 capillary electrophoresis system using a 50 cm × 50 μm ID bare fused-silica capillary (Beckman). Phosphate buffer (50 mM, pH 8.0) was used as running buffer. Electrophoresis was started at 15 kV and 20 ± 0.1°C. Samples were injected at 10 kV for 20 s and detected at 214 nm. Prior to use, the capillary was pre-treated successively with 0.1 M NaOH for 60 min, water for 30 min, and finally with running buffer until the baseline becomes smooth. Between runs the capillary was washed sequentially with 0.1 M NaOH, water, and running buffer for 4 min each. Solutions were filtered through a 0.22 μm PTFE membrane prior to use. To ensure proper folding of the TAR RNA structure, the RNA solutions were annealed by heating for 3 min at 95°C and cooled slowly. TAR-compound or TAR-Tat peptide complex was incubated for 30 min at 4°C (binding buffer: 10 mM Tris-HCl, 70 mM NaCl, 0.2 mM EDTA, and 5% glycerol, pH 7.4) before CE analysis.
16. The samples were prepared with a wide range of Tat (or the compounds) diluted by binding buffer (10 mM Tris-HCl, 70 mM NaCl, 0.2 mM EDTA, and 5% glycerol, pH 7.4) and CE procedure was as described in Ref. 15. All experiments were in triplicate. Relative standard deviation (RSD) was calculated from a series of three experiments carried out with the same sample in 1 day. Linear equations of the concentration and the peak area were calculated. With the peak areas of free Tat or the free compounds were obtained as described in Ref. 15, their concentration in the mixture could be calculated.
17. 2 containing humidified air. The cells were seeded at a six-well plate 24 h prior to transfection which was performed by standard calcium phosphate co-precipitation techniques with optimum amounts of the plasmids pLTRCAT and pSVCMVTAT. Twenty four hours later, the culture medium was removed and the cells were washed twice with phosphate-buffered saline (PBS). Then the transfected cells were added to fresh medium together with diluted compounds of final concentration 30 μM, respectively, and incubated for another 24 h. After forty eight hours post-transfection, the cells were harvested and analyzed for CAT activity using a commercial CAT ELISA kit (Roche Molecular Biochemicals) in accordance with the manufacturer's protocol. All data were reported as a percentage of CAT activity (±SD). Results shown are representative of three independent experiments.
18. 50 was defined as the compound concentration required to protect cells against the cytopathogenicity of SIV by 50%.