Hapten Optimization for Cocaine Vaccine with Improved Cocaine Recognition

• Published in: Chemical biology & drug design Vol. 84; no. 3; pp. 354 - 363
• Main Authors: Ramakrishnan, Muthu, Kinsey, Berma M., Singh, Rana A., Kosten, Thomas R., Orson, Frank M.
• Format: Journal Article
• Language: English
• Published: England Blackwell Publishing Ltd 01-09-2014
• Subjects: addiction; Animals; Antibodies - analysis; Antibodies - immunology; Antibodies - metabolism; antibody; benzoylecgonine; carrier protein; cocaine; Cocaine - analogs & derivatives; Cocaine - chemistry; Cocaine - immunology; Enzyme-Linked Immunosorbent Assay; hapten; Haptens - chemistry; Haptens - immunology; Male; Mice; Mice, Inbred BALB C; outer membrane protein complex; specificity; substance abuse; TA-CD vaccine; vaccine; Vaccines - immunology; vaccine; antibody; cocaine; hapten; outer membrane protein complex; addiction; specificity; TA-CD vaccine; carrier protein; substance abuse; benzoylecgonine; mice
• ISSN: 1747-0277; 1747-0285
• DOI: 10.1111/cbdd.12326

In the absence of any effective pharmacotherapy for cocaine addiction, immunotherapy is being actively pursued as a therapeutic intervention. While several different cocaine haptens have been explored to develop anticocaine antibodies, none of the hapten was successfully designed, which had a protonated tropane nitrogen as is found in native cocaine under physiological conditions, including the succinyl norcocaine (SNC) hapten that has been tested in phase II clinical trials. Herein, we discuss three different cocaine haptens: hexyl norcocaine (HNC), bromoacetamido butyl norcocaine (BNC), and succinyl butyl norcocaine (SBNC), each with a tertiary nitrogen structure mimicking that of native cocaine which could optimize the specificity of anticocaine antibodies for better cocaine recognition. Mice immunized with these haptens conjugated to immunogenic proteins produced high titre anticocaine antibodies. However, during chemical conjugation of HNC and BNC haptens to carrier proteins, the 2β methyl ester group is hydrolyzed, and immunizing mice with these conjugate vaccines in mice produced antibodies that bound both cocaine and the inactive benzoylecgonine metabolite. While in the case of the SBNC conjugate, vaccine hydrolysis of the methyl ester did not appear to occur, leading to antibodies with high specificity to cocaine over BE. Although we observed similar specificity with a SNC hapten, the striking difference is that SBNC carries a positive charge on the tropane nitrogen atom, and therefore, it is expected to have better binding of cocaine. The 50% cocaine inhibitory concentration (IC50) value for SBNC antibodies (2.8 μm) was significantly better than the SNC antibodies (9.4 μm) when respective hapten–BSA was used as a substrate. In addition, antibodies from both sera had no inhibitory effect from BE. In contrast to BNC and HNC, the SBNC conjugate was also found to be highly stable without any noticeable hydrolysis for several months at 4 °C and 2–3 days in pH 10 buffer at 37 °C.
Optimization of cocaine conjugate vaccine with improved hapten design with a tertiary nitrogen structure mimicking that of physiological structure of cocaine. The hapten conjugates produced antibodies with better cocaine recognition than our current generation TA‐CD vaccine. In addition, we have also shown a new approach to study the stability of cocaine hapten‐conjugates by exploring the unique specificity of commercially available anti‐cocaine antibodies.