An efficient biodelivery system for antisense polyamide nucleic acid (PNA).
With the aim of developing a general and straightforward procedure for the intracellular delivery of naked peptide nucleic acids (PNAs), we designed an intracellularly biodegradable triphenylphosphonium (TPP) cation based transporter system. In this system, TPP is linked, via a biolabile disulfide bridge, to an activated mercaptoethoxycarbonyl moiety, allowing its direct coupling to the N-terminal extremity of a free PNA through a carbamate bond. We found that such TPP-PNA-carbamate conjugates were highly stable in a cell culture medium containing fetal calf serum. In a glutathione-containing medium mimicking the cytosol, the conjugates were rapidly degraded into an unstable intermediate, which spontaneously decomposed, releasing the free PNA. Using a fluorescence-labeled PNA-TPP conjugate, we demonstrated that conjugates were taken up by cells. Efficient cellular uptake and release of the PNA into the cytosol was further confirmed by the anti-HIV activity measured for the TPP-conjugate of a 16-mer PNA targeting the TAR region of the HIV-1 genome. This conjugate exhibited an IC(50) value of 1 microM, while the free 16-mer PNA did not inhibit replication of HIV in the same cellular test.
Mehiri
Mohamed
M
Laboratoire de Chimie des Molécules Bioactives et des Arômes, Université de Nice-Sophia Antipolis-CNRS, Institut de Chimie de Nice, Nice, France.
Upert
Gregory
G
Tripathi
Snehlata
S
Di Giorgio
Audrey
A
Condom
Roger
R
Pandey
Virendra N
VN
Patino
Nadia
N
D019380:Anti-HIV Agents / Q000378:metabolism / Q000494:pharmacology; D016375:Antisense Elements (Genetics); D002460:Cell Line; D003470:Culture Media; D016503:Drug Delivery Systems; D016325:HIV Long Terminal Repeat / Q000187:drug effects; D015497:HIV-1 / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D006801:Humans; D009757:Nylons / Q000737:chemistry / Q000378:metabolism*; D020135:Peptide Nucleic Acids / Q000138:chemical synthesis / Q000378:metabolism* / Q000494:pharmacology; D014779:Virus Replication / Q000187:drug effects
D019380:Anti-HIV Agents; D016375:Antisense Elements (Genetics); D003470:Culture Media; D009757:Nylons; D020135:Peptide Nucleic Acids