Synthesis of 4-[2-aminoethyl(nitrosamino)]-1-pyridin-3-yl-butan-1-one, a new NNK hapten for the induction of N-nitrosamine-specific antibodies.
4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is one of the most abundant and potent procarcinogens in tobacco smoke. In order to induce a strong and substained antibody response against NNK, we developed a functionalized derivative that closely mimicked its structural features, in particular, the pyridyloxobutyl moiety, the adjacent ketone, and the N-nitrosamino group. This hapten was conjugated via a C 2 linker to the highly immunogenic diphteria toxoid licensed as a vaccine in humans to induce polyclonal and monoclonal antibodies. Two monoclonal antibodies were obtained with Kd values of 45.8 nM (P9D5) and 37.6 nM (P7H3), respectively, for NNK-C 2. Both the monoclonal (P9D5 and P7H3) and polyclonal antibodies reacted strongly with NNK (IC 50 = 80 microM or 160 microM) and NNAL (IC 50 = 29 microM or 93 microM) and to a lesser extent with some of the metabolites of NNK. Interestingly, the mAbs did not react with the metabolites of the detoxification pathways such as NNK-N-Oxide and NNAL-N-Oxide (IC 50 > 512 microM). Therefore, such antibodies detect NNK and NNAL and may have the potential to modulate their redistribution in vivo, perhaps reducing some detrimental effects of smoking.
Prodhomme
Emmanuel J F
EJ
Institute of Immunology, Laboratoire National de Santé, 20A rue Auguste Lumière, L-1011, Luxembourg, Luxembourg.
Ensch
Corinne
C
Bouche
Fabienne B
FB
Kaminski
Thomas
T
Deroo
Sabrina
S
Seck
Pierre
P
Kirsch
Gilbert
G
Muller
Claude P
CP
D000906:Antibodies / Q000097:blood / Q000276:immunology*; D002352:Carrier Proteins / Q000737:chemistry / Q000276:immunology; D006241:Haptens / Q000737:chemistry*; D015394:Molecular Structure; D009602:Nitrosamines / Q000138:chemical synthesis* / Q000737:chemistry* / Q000276:immunology*
C525967:4-(2-aminoethyl(nitrosamino))-1-pyridin-3-yl-butan-1-one; D000906:Antibodies; D002352:Carrier Proteins; D006241:Haptens; D009602:Nitrosamines; C016583:4-(N-methyl-N-nitrosamino)-1-(3-pyridyl)-1-butanone