Viral Hepatitis
Copyright ©The Author(s) 2003. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Jun 15, 2003; 9(6): 1251-1255
Published online Jun 15, 2003. doi: 10.3748/wjg.v9.i6.1251
Distribution and anti-HBV effects of antisense oligodeoxynu-cleotides conjugated to galactosylated poly-L-lysine
Su-Jun Zheng, Sen Zhong, Jian-Jun Zhang, Feng Chen, Hong Ren, Cun-Liang Deng
Su-Jun Zheng, Jian-Jun Zhang, Hong Ren, Institute of Viral Hepatitis, Chongqing University of Medical Sciences, Chongqing 400016, China
Sen Zhong, Feng Chen, Cun-Liang Deng, Department of Infectious Diseases, Luzhou Medical College Hospital, Luzhou 646000, Sichuan Province, China
Author contributions: All authors contributed equally to the work.
Supported by the Natural Science Foundation of China, No. 39370648
Correspondence to: Sen Zhong, Department of Infectious Diseases, Luzhou Medical College Hospital, Luzhou 646000, China.
Telephone: +86-830-2392712-5419
Received: December 8, 2002
Revised: January 4, 2003
Accepted: January 8, 2003
Published online: June 15, 2003

AIM: To describe distribution of the phosphorothioated antisense oligodeoxynucleotides (PS-asODNs) conjugated to galactosylated poly-L-lysine (Gal-PLL) in mice, and to observe their effects on expression of HBV gene in the 2.2.15 cells and transgenic mice.

METHODS: According to the result of direct sequencing of PCR amplified products, a 16 mer phosphorothioate analogue of the antisense oligodeoxynucleotides (PS-asODNs) directed against the HBV U5-like region was conjugated to the hepatotropic Gal-PLL molecules. Its distribution was demonstrated using asODNs labeled with 32P at the 5’ terminus with a T4-polynucleotide Kinase. Its inhibition effect on HBV expression was observed in the transfected 2.2.15 cells and transgenic mice.

RESULTS: The Gal-PLL and asODNs could form stable complex at a molar ratio of 2:1. As shown in the HBV-transfected 2.2.15 cells, the inhibition effects of asODNs alone and asODNs conjugated to Gal-PLL, at 10 μmol/L for both, on HBsAg and HBeAg production were different, the former being 70% and 58%, respectively, and the latter being 96% and 82%, respectively. A more pronounced reduction was also observed in viral DNA load in the culture supernatant for the test with Gal-PLL-asODNs. Among many mouse organs, livers retained more asODNs molecules after administration. The preferential concentration in liver was found to be 52.14% for Gal-PLL-asODNs, as high as 2.38-fold of that for asODNs (21.9%). Both elements decreased gradually in liver, with 2.9% of the former, 5.99% of the latter retained 24 h after the administration. The injection interval, therefore, was recommended to be 24 h. In the transgenic mice, serum HBsAg decreased significantly (P < 0.01) at the 12th day after administrating Gal-PLL- asODNs, the serum HBV DNA turned negative in 4 of the 6 mice.

CONCLUSION: Antisense oligodeoxynucleotides conjugated to Gal-PLL can be concentrated in liver and intaked by hepatocytic cells. This may result in specific inhibition of expression and replication of HBV in vitro and in vivo.

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