RESUMO
A method for potentiating the response to an anti-cocaine vaccine by leveraging xenoreactive antibodies against the carbohydrate epitope Galα1,3-Gal (GAL) was found to result in a highly specific anti-cocaine response that was able to significantly attenuate cocaine-induced locomotion at 20 mg kg-1 with superior efficacy compared to a standard conjugate.
Assuntos
Anticorpos/imunologia , Anticorpos/farmacologia , Cocaína/antagonistas & inibidores , Dissacarídeos/antagonistas & inibidores , Dissacarídeos/imunologia , Locomoção/efeitos dos fármacos , Animais , Anticorpos/química , Cocaína/farmacologia , Relação Dose-Resposta a Droga , Camundongos , Conformação MolecularRESUMO
Methamphetamine (MA) addiction is a serious public health problem, and current methods to abate addiction and relapse are currently ineffective for mitigating this growing global epidemic. Development of a vaccine targeting MA would provide a complementary strategy to existing behavioral therapies, but this has proven challenging. Herein, we describe optimization of both hapten design and formulation, identifying a vaccine that elicited a robust anti-MA immune response in mice, decreasing methamphetamine-induced locomotor activity.
Assuntos
Haptenos/imunologia , Metanfetamina/imunologia , Vacinas/imunologia , Animais , Formação de Anticorpos , Ensaio de Imunoadsorção Enzimática , Locomoção/efeitos dos fármacos , Metanfetamina/farmacologia , Camundongos , Vacinas/químicaRESUMO
Fentanyl is an addictive prescription opioid that is over 80â times more potent than morphine. The synthetic nature of fentanyl has enabled the creation of dangerous "designer drug" analogues that escape toxicology screening, yet display comparable potency to the parent drug. Alarmingly, a large number of fatalities have been linked to overdose of fentanyl derivatives. Herein, we report an effective immunotherapy for reducing the psychoactive effects of fentanyl class drugs. A single conjugate vaccine was created that elicited high levels of antibodies with cross-reactivity for a wide panel of fentanyl analogues. Moreover, vaccinated mice gained significant protection from lethal fentanyl doses. Lastly, a surface plasmon resonance (SPR)-based technique was established enabling drug-specificity profiling of antibodies derived directly from serum. Our newly developed fentanyl vaccine and analytical methods may assist in the battle against synthetic opioid abuse.
Assuntos
Analgésicos Opioides/toxicidade , Antídotos/uso terapêutico , Drogas Desenhadas/síntese química , Fentanila/toxicidade , Vacinas/administração & dosagem , Animais , Fentanila/farmacocinética , Camundongos , Distribuição TecidualRESUMO
Full details of studies are disclosed on the total syntheses of binding pocket analogues of vancomycin bearing the peripheral L-vancosaminyl-1,2-D-glucosyl disaccharide that contain changes to a key single atom in the residue-4 amide (residue-4 carbonyl O â S, NH, H2) designed to directly address the underlying molecular basis of resistance to vancomycin. Also disclosed are studies piloting the late-stage transformations conducted on the synthetically more accessible C-terminus hydroxymethyl aglycon derivatives and full details of the peripheral chlorobiphenyl functionalization of all of the binding-pocket-modified vancomycin analogues designed for dual D-Ala-D-Ala/D-Ala-D-Lac binding. Their collective assessment indicates that combined binding pocket and chlorobiphenyl peripherally modified analogues exhibit a remarkable spectrum of antimicrobial activity (VSSA, MRSA, and VanA and VanB VRE) and impressive potencies against both vancomycin-sensitive and vancomycin-resistant bacteria (MICs = 0.06-0.005 and 0.5-0.06 µg/mL for the amidine and methylene analogues, respectively) and likely benefit from two independent and synergistic mechanisms of action, only one of which is dependent on D-Ala-D-Ala/D-Ala-D-Lac binding. Such analogues are likely to display especially durable antibiotic activity that is not prone to rapidly acquired clinical resistance.
Assuntos
Antibacterianos/síntese química , Antibacterianos/farmacologia , Compostos de Bifenilo/química , Vancomicina/síntese química , Vancomicina/farmacologia , Antibacterianos/química , Antibacterianos/metabolismo , Bactérias/efeitos dos fármacos , Bactérias/metabolismo , Sítios de Ligação , Técnicas de Química Sintética , Farmacorresistência Bacteriana , Testes de Sensibilidade Microbiana , Relação Estrutura-Atividade , Vancomicina/química , Vancomicina/metabolismoRESUMO
Autoinducer-2 (AI-2)-mediated quorum sensing (QS) is utilised for both intra- and inter-species communication by a wide variety of bacteria. An understanding of the mechanism of this communication has the potential to elucidate new targets for antibacterial therapeutics. Herein, we report the synthesis of DPD analogues with modified dynamic equilibria and the evaluation of their behaviour in Gram-negative bacteria. None of the compounds showed modulation of QS in S. Typhimurium, and although no antagonism of V. harveyi was observed, chloro-analogue C5-Cl-DPD showed modest agonism in this marine bacterium. This raises the possibility that access to a cyclic form of DPD may not be required for AI-2-mediated QS in V. harveyi.
RESUMO
Cocaine abuse is problematic, directly and indirectly impacting the lives of millions, and yet existing therapies are inadequate and usually ineffective. A cocaine vaccine would be a promising alternative therapeutic option, but efficacy is hampered by variable production of anticocaine antibodies. Thus, new tactics and strategies for boosting cocaine vaccine immunogenicity must be explored. Flagellin is a bacterial protein that stimulates the innate immune response via binding to extracellular Toll-like receptor 5 (TLR5) and also via interaction with intracellular NOD-like receptor C4 (NLRC4), leading to production of pro-inflammatory cytokines. Reasoning that flagellin could serve as both carrier and adjuvant, we modified recombinant flagellin protein to display a cocaine hapten termed GNE. The resulting conjugates exhibited dose-dependent stimulation of anti-GNE antibody production. Moreover, when adjuvanted with alum, but not with liposomal MPLA, GNE-FliC was found to be better than our benchmark GNE-KLH. This work represents a new avenue for exploration in the use of hapten-flagellin conjugates to elicit antihapten immune responses.
Assuntos
Cocaína/imunologia , Flagelina/química , Haptenos/química , Vacinas/imunologia , Adjuvantes Imunológicos/química , Compostos de Alúmen/química , Animais , Peso Corporal , Ensaio de Imunoadsorção Enzimática , Flagelina/imunologia , Haptenos/imunologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , RadioimunoensaioRESUMO
A leading nicotine conjugate vaccine was only efficacious for one-third of clinical trial participants, likely due in part to its use of racemic nicotine hapten, (±)-3'-AmNic. Immunization of male Wistar rats with (+)-, (-)-, or (±)-3'-AmNicSucTT and subsequent antibody immunoassays suggest that a vaccine using enantiopure (-)-3'-AmNic hapten imparts superior capacity to bind (-)-nicotine. Future nicotine vaccine clinical candidates must incorporate this design consideration (i.e., hapten enantiopurity) in order to maximize efficacy.
Assuntos
Haptenos/imunologia , Nicotina/imunologia , Vacinas Conjugadas/imunologia , Animais , Ensaio de Imunoadsorção Enzimática , Masculino , Nicotina/metabolismo , Ratos , Ratos Wistar , EstereoisomerismoRESUMO
Studies on the further development of the sequential glycosylations of the vancomycin aglycon catalyzed by the glycosyltransferases GtfE and GtfD and the observation of unusual, perhaps unexpected, aglycon substrate substituent effects on the rate and efficiency of the initial glycosylation reaction are reported.
Assuntos
Antibacterianos/síntese química , Vancomicina/síntese química , Antibacterianos/química , Glucosiltransferases/metabolismo , Glicosilação , Estrutura Molecular , Vancomicina/químicaRESUMO
Vaccines for drugs of abuse have yet to achieve full clinical relevance, largely due to poor/inconsistent immune responses in patients. The use of multivalent scaffolding as a means to tailor drug-hapten density and clustering was examined in the context of drug-immune response modulation. A modular trivalent hapten containing a diglycine spacer, triAM1(Gly)2, was synthesized and shown to elicit anti-nicotine antibodies at equivalent affinity and concentration to the monovalent AM1 analog, despite in this instance having a lower effective hapten density. Augmenting this data, the corresponding monovalent hapten AM1(Gly)2 resulted in enhanced antibody affinity and concentration. Drug-hapten clustering represents a new vaccine paradigm, and, while examined only in the context of nicotine, it should be readily translatable to other drugs of abuse.