Epibatidine

9/3/11 5:22 PM

Epibatidine
Matthew J. Dowd , Graduate Student Department of Medicinal Chemistry Virginia Commonwealth University Richmond, VA 23298-0540 USA Click here for the CHIME-enhanced version of this article. Also, click on any structure to view a 3-D version of the molecule. Within the rainforest of Ecuador resides a small, colorful, seemingly harmless amphibian called Epipedobates tricolor ( Figure 1 ). This frog first introduced itself to the scientific world in 1974. It was then that Dr. John Daly of the National Institutes of Health isolated from the frog a compound initially called alkaloid 208/210 (its MW from mass spectrometry) [1]. Daly demonstrated that this new alkaloid was a potent analgesic (as measured in the Straub-tail response when injected into mice). Even after subsequent trips to South America, too little of the compound was isolated to make a structural determination.[2,3] Because of this lack of compound, for both scientific and political reasons, the remaining sample, about 750 micrograms, was kept in storage for several years. During the early 1990's, when NMR instruments and methods became more sensitive and sophisticated, Daly's group was able to determine the structure of alkaloid 208/210, which was renamed epibatidine ( 1 )(1 R, 2 R, 4 S exo-2-(6-chloro-3-pyridyl)-7-azabicyclo[2.2.1]heptane). As stated above, epibatidine was shown to be a potent analgetic (about 200 times more potent than morphine). The truly exciting discovery was that epibatidine's mechanism of action appeared to be non-opioid. Many potent pain relieving drugs are opiates, morphine ( 2 ) being a very familiar example. Morhpine is an effective and potent analgesic; however, the potential for addiction and the development of morphine tolerance are major drawbacks to its use. Several major pharmaceutical companies have focused their efforts on discovering better analgesics. When Daly showed that epibatidine's effect was not blocked by naloxone, an opioid antagonist, this revelation produced much enthusiasm in the hope for a better drug.

If epibatidine did not exert its analgesic effect through opioid receptors, how then did it produce the pain relief? Shortly after the publication of the structure of epibatidine, several research groups, including Daly's, determined the answer by examining epibatidine's interaction with nicotinic acetylcholine receptors (AChRs), a type of
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4 S -(-)-Epibatidine is the natural stereoisomer excreted by the frog. sparked a resurgence in the medicinal chemistry of nicotine and nicotinic analogues.5 A [12]. Beers and Reich [9]. the chirality.vcu. although not all. protonated basic nitrogen).1-5.html Page 2 of 6 . it did so at extremely low concentrations (Ki=0. This modeling experiment was the first to show in a three dimensional fashion that epibatidine and nicotine may interact with similar receptor features. pyridine N or carbonyl O) and a center of positive charge (e. several research groups have proposed pharmacophores for the nicotinic receptor. the models all contain a hyrogen bond acceptor atom (e.hence their name. Barlow and Johnson [10]. S -(-)-nicotine (Ki = 1-2 nM). For example. With the emergence of epibatidine as a high affinity nicotinic agonist. whereas the enantiomers of epibatidine occupy the same molecular volume. Medicinal chemists have attempted to define the structural and chemical features that are important for epibatidine's high affinity. or absolute spatial configuration of the molecule.8]. one goal of medicinal chemists is to define a pharmacophore the optimal three dimensional arrangement of chemical and structural molecular features required by a certain receptor. This distance is often referred to as the "internitrogen distance" because most. along with previous research illustrating potentially beneficial effects of nicotine [7.043-0. Its enantiomer has been synthesized. both basic nitrogens are part of a five-membered ring (in epibatidine. 2 R. With some simplification.g. Not only did epibatidine bind to and activate these receptors.Epibatidine 9/3/11 5:22 PM ligand-gated ion channel whose endogenous ligand is acetylcholine ( 3 ) [4-6]. Both contain a six-membered pyridine ring. rationalized the nonstereospecificity of epibatidine [5]. chlorine is green. Another early insight into the binding of epibatidine resulted from a comparison of the structures of nicotine and epibatidine. tested for receptor affinity. the naturally occurring stereoisomer. With many biologically active compounds. separated by a distance of approximately 4.8 A.. and shown to have the same affinity as the natural isomer.edu/medchem/articles/epi/index. showed that energy-minimized molecular models of the compounds could be overlayed such that major structural features are in similar positions in space ( Figure 2 ) [5]. both contain a basic nitrogen linked to the pyridine ring by one or two carbons. S -(-)-Nicotine ( 4 ) also activates these receptors . a research group at Abbott Laboratories pubished work in which they synthesized a series of pyridyl ether compounds which are nicotinic http://www. often influences its activity. has about 20-fold higher affinity than its enantiomer. 1 R. The enantiomers of nicotine appear to occupy different volumes in spaces. In the past. Glennon and his group reevaluated the nicotinic pharmacophore and produced a model which indicated an optimal internitrogen distance of 5. Dukat et al . Fig 2.055 nM or about 55 pM).. nicotinic analogues contain a pyridine nitrogen and a more basic nitrogen. and Sheridan and coworkers [11] have suggested nicotine pharmacophores. the five membered ring is part of the azabicycloheptane structure).pharmacy. R-(+)-nicotine ( 4 ) (Ki = 25 nM). In 1996. The finding that the analgesic effects of epibatidine are blocked by mecamylamine (a noncompetitive nicotinic antagonist). When studying biologically active compounds. Nitrogens are blue. In fact. A molecular modeling study by Dukat et al . Superposition of nicotine (cyan) and epibatidine (red).g.

the most studied ligand-gated ion channel. some as potent as epibatidine. is composed of ( 1)2 1 . which utilized an -iodocycloalkenone in a modified Stille reaction [31].0 4 4 84. The results are shown in Table 1. Broka [25].vcu. epiboxidine is 20-fold less toxic than epibatidine. Aoyagi reported a total sythesis in which the key reaction was an asymmetric Diels-Alder reaction with a chiral N-acylnitroso compound as the dieneophile [30].. there is a somewhat significant change in affinity when the subunits are changed (e. In addition. Sirisoma and Johnson described their synthetic route. The binding affinity and the pharmacological effects of a particular ligand are dependent upon the subunit composition of the nAChR. Daly used this same isosteric replacement in epibatidine. a hybrid between epibatidine and ABT-418 ( 5 ) [32]. ABT-418 was designed by replacing the pyridine ring of nicotine with a methylisoxazole ring. and nicotinic receptors [7. Corey [23]. With such a brief overview.edu/medchem/articles/epi/index.Y.1 A may be optimal for interaction at the nicotinic receptor [13]. it is impossible to review or mention all the research and scientists that have contributed to our understanding of epibatidine.8. chemists are certain to devise additional avenues to its synthesis. Because of the rather simple but intriguing structure of epibatine. an isosteric analogue of nicotine. There are.7 Synthetic organic chemists have also shown intense interest in epibatidine. Luetje's lab. For more details. please refer to any of the excellent reviews of epibatidine[15-17]. E. Molecular modeling studies incorporating these new agents suggested that a internitrogen distance closer to 6. One area of complexity concerns receptor subtypes and populations. Neuronal nAChRs are composed of various combinations of and subunits. As can be observed.22]. producing epiboxidine. Epibatidine's azabicycloheptane system is not common in natural products. is a ligand-gated ion channel. Also. epiboxidine (Ki = 0. The nicotinic acetylcholine receptor. each with a different composition of subunits. However.9) and four beta ( 1 . 30-fold difference in affinity when 3 2 is changed to 3 4). Although not as potent as epibatidine. and different pharmacological properties.J.Epibatidine 9/3/11 5:22 PM agonists. To date. nicotinic ligands [18-20].html Page 3 of 6 .8 3 2 13. and (epsilon). Chen [24]. the affinity of epibatidine for several different subtypes of the nAChR was reported [14].pharmacy. new analogues of epibatidine have been and are still being synthesized. many of the complexities of the chemistry and biology have been omitted for brevity's sake.6 3 4 303 4 2 30. and Clayton and Regan [26] were the among the first to report total syntheses of epibatidine. (beta). http://www. composed of five individual subunits: (alpha). So what does the future hold for epibatidine? The chance of epibatidine ever being used as a medicinal agent is quite low because of its high toxicity.01 nM) and ABT-418 (Ki = 10 nM).3 2 4 86. Also. One interesting analogue is epiboxidine ( 6 ). (delta). There is still much work to be done before a precise nicotine pharmacophore can be agreed upon by the medicinal chemists. Table 1. Receptor Affinity (pM) 2 2 10. More recently. has analgesic and cognitive-enhancing properties in certain test systems. In a recent report from Dr. nine alpha subunits ( 1 . as stated above. several different subtypes of receptor. replacing the chloropyridine ring with the methylisoxazole ring. Many other synthetic routes were later reported (See references 27-29 for reviews).6 nM) has higher affinity to the nAChR than nicotine (Ki = 1. however.g. The muscle type nAChR.4) subunits have been discovered.21. T. (gamma). ABT-418. Epibatidine Affinity at Neuronal nAChR subtypes.

The diazabicyclic pyrazine DBO-83 ( 10) is another high affinity (Ki = 4 nM) nicotinic ligand that has some structural similarity to epibatidine [37. compound 7 was shown to have analgesic potency comparable to that of epibatidine [34].. 2) Badio. Panel. Li. Another puzzle to solve is the source of epibatidine. The common presumption is that the frog obtains epibatidine. identifying the producer of this potent nicotinic agonist may provide an abundant source of epibatidine.. 1993.pharmacy.F. Soc. However. Dumas. 250 . have been synthesized and tested. Res.. The idea for this compound originated partly from research aimed at discovering analgesics that were selective for the mu-opioid receptor. J. Chem. Spande. 114 .edu/medchem/articles/epi/index.. Daly. Thirty Years of Discovering Arthropod Alkaloids in Amphibian Skin. L. J. and the azabicyclooctane analogue 9 [33-36]. T. H. 563569. Whichever the answer. Am. he could not isolate or detect any epibatidine[2. Garraffo. T. M. 1994.M. Prod.L. Damaj. Edwards.html Page 4 of 6 . Date posted: 1/13/99 References 1) Spande. because of the structural similarity of epibatidine and nicotine.. 131-139. 440-448. bis -homoepibatidine ( 8 ). Mol.. 1998. Epibatidine: A Novel (Chloropyridyl)azabicycloheptane with Potent Analgesic Activity from an Ecuadoran Poison Frog. Nat. Res. 1994.C. 3) Daly. Ip.. Med. Biftu.vcu.W. Pharmacol. Glennon. Eckman.M. B. On the other hand. Eur.F. tricolor frogs in captivity..W..J. R. http://www. B. 61. C. Interestingly.Y.Epibatidine 9/3/11 5:22 PM Several analogues of epibatidine. T. Med. W. M. a plant-derived alkaloid. Martin.A. D.I. Epibatidine: A Very High Affinity Nicotinic Receptor Ligand. J. a Potent Analgetic and Nicotinic Agonist.. Chem. or some biological precursor. 162172. 1992. J. J. Chem. Epibatidine is a Nicotinic Analgesic.. Daly. J. 3475-3478.. Yeh. when Daly raised some E. Epibatidine: Discovery and Definition as a Potent Analgesic and Nicotinic Agonist.W. Shen.. Glassco. J. 4) Badio.. These include homoepibatidine ( 7 ). .. H. Insects are one suspected source. a floral source may be a possibility.. 4 . Garraffo.3]. 1994. Researchers first thought that the frog produced the compound biochemically.. 6) Qian.W. 38]. 5) Dukat. 45. E. Pharmacol. L.. J.. S. Libertine-Garahan. M. Daly. from a dietary source. T. May. in which the azabicycloheptane ring has been altered. H.R. B.W. R13-R14. 4 . Epibatidine.. T.

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