Synthesis, conformational analysis and CB1 binding affinity of hairpin-like anandamide pseudopeptide mimetics
Maria Di Marzo
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, Italy
Search for more papers by this authorAgostino Casapullo
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, Italy
Search for more papers by this authorGiuseppe Bifulco
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, Italy
Search for more papers by this authorPaola Cimino
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, Italy
Search for more papers by this authorAlessia Ligresti
Endocannabinoid Research Group, Istituto di Chimica Biomolecolare, CNR, 80078, Pozzuoli, NA, Italy
Search for more papers by this authorVincenzo Di Marzo
Endocannabinoid Research Group, Istituto di Chimica Biomolecolare, CNR, 80078, Pozzuoli, NA, Italy
Search for more papers by this authorRaffaele Riccio
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, Italy
Search for more papers by this authorCorresponding Author
Luigi Gomez-Paloma
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, Italy
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, ItalySearch for more papers by this authorMaria Di Marzo
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, Italy
Search for more papers by this authorAgostino Casapullo
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, Italy
Search for more papers by this authorGiuseppe Bifulco
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, Italy
Search for more papers by this authorPaola Cimino
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, Italy
Search for more papers by this authorAlessia Ligresti
Endocannabinoid Research Group, Istituto di Chimica Biomolecolare, CNR, 80078, Pozzuoli, NA, Italy
Search for more papers by this authorVincenzo Di Marzo
Endocannabinoid Research Group, Istituto di Chimica Biomolecolare, CNR, 80078, Pozzuoli, NA, Italy
Search for more papers by this authorRaffaele Riccio
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, Italy
Search for more papers by this authorCorresponding Author
Luigi Gomez-Paloma
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, Italy
Dipartimento di Scienze Farmaceutiche, Università di Salerno, 84084 Fisciano, Salerno, ItalySearch for more papers by this authorAbstract
We have designed, synthesized and evaluated the CB1 binding affinity of a number of new conformationally restricted lipopeptides (1–17). All of them present some of the AEA key structural elements incorporated in a hairpinlike peptide framework. Among them, compounds 1–3 and 8 showed CB1 affinities in competitive binding assays with Ki values in the micromolar range (Ki of AEA = 0.8 µM in the same assay). The remaining pseudopeptides showed little binding to the CB1 receptor (with Ki values ≥ 50 µM). Conformational analysis on two representative compounds, performed by a combination of NMR studies, restrained molecular dynamics and QM calculations, allowed us to shed light on the structure-activity relationships (SAR), pointing to a correlation between the predominance of the hairpinlike structural motif and the CB1 binding affinity. In a more general context, the present study may also prove useful in gaining additional insight into the biological relevance of the various AEA conformations. Copyright © 2006 European Peptide Society and John Wiley & Sons, Ltd.
REFERENCES
- 1Gaoni Y, Mechoulam R, Hashish III. Isolation, structure, and partial synthesis of an active constituent of hashish. J. Am. Chem. Soc. 1964; 86: 1646–1647.
- 2Devane WA, Hanus L, Breuer A, Pertwee RG, Stevenson LA, Griffin G, Gibson D, Mandelbaum A, Etinger A, Mechoulam R. Isolation and structure of a brain constituent that binds to the cannabinoid receptor. Science 1992; 258: 1946–1949.
- 3Martin BR. Cellular effects of cannabinoids. Pharmacol. Rev. 1986; 38: 45–74.
- 4Dewey WL. Cannabinoid pharmacology. Pharmacol. Rev. 1986; 38: 151–178.
- 5Razdan RK. The total synthesis of cannabinoids. Total Syntheses in Natural Products, J Apsimon (ed), Wiley VCH, New York, 1981; 4 185–262.
10.1002/9780470129678.ch2 Google Scholar
- 6Razdan RK. Structure-activity relationships in cannabinoids. Pharmacol. Rev. 1986; 38: 75–149.
- 7Khanolkar AD, Makriyannis A. Structure-activity relationships of anandamide, an endogenous cannabinoid ligand. Life Sci. 1999; 65: 607–616.
- 8Bisogno T, Maurelli S, Melck D, De Petrocellis L, Di Marzo V. Biosynthesis, uptake, and degradation of anandamide and palmitoylethanolamide in leukocytes. J. Biol. Chem. 1997; 272: 3315–3323.
- 9Di Marzo V, De Petrocellis L, Bisogno T, Maurelli S. The endogenous cannabimimetic eicosanoid anandamide induces arachidonate release in J774 mouse macrophages. Adv. Exp. Med. Biol. 1997; 407: 341–346.
- 10Cabral GA, Toney DM, Fisher-Stenger K, Harrison MP, Marciano-Cabral F. Anandamide inhibits macrophage-mediated killing of tumor necrosis factor-sensitive cells. Life Sci. 1995; 56: 2065–2072.
- 11Schwarz H, Blanco FJ, Lotz M. Anandamide, an endogenous cannabinoid receptor agonist inhibits lymphocyte proliferation and induces apoptosis. J. Neurochem. 1994; 55: 107–115.
- 12Ishac EJ, Jiang L, Lake KD, Varga K, Abood ME, Kunos G. Inhibition of exocytotic noradrenaline release by presynaptic cannabinoid CB1 receptors on peripheral sympathetic nerves. Br. J. Pharmacol. 1996; 118: 2023–2028.
- 13Pugh G Jr, Smith PB, Dombrowski DS, Welch SP. The role of endogenous opioids in enhancing the antinociception produced by the combination of delta 9-tetrahydrocannabinol and morphine in the spinal cord. J. Pharmacol. Exp. Ther. 1996; 279: 608–616.
- 14Martin BR, Compton DR, Thomas BF, Prescott WR, Little PJ, Razdan RK, Johnson MR, Melvin LS, Mechoulam R, Ward SJ. Behavioral, biochemical, and molecular modeling evaluations of cannabinoid analogs. Pharmacol., Biochem. Behav. 1991; 40: 471–478.
- 15Compton DR, Johnson MR, Melvin LS, Martin BR. Pharmacological profile of a series of bicyclic cannabinoid analogs: classification as cannabimimetic agents. J. Pharmacol. Exp. Ther. 1992; 260: 201–209.
- 16Adams IB, Ryan W, Singer M, Razdan RK, Compton DR, Martin BR. Pharmacological and behavioral evaluation of alkylated anandamide analogs. Life Sci. 1995; 56: 2041–2048.
- 17Khanolkar AD, Abadji V, Lin S, Hill WA, Taha G, Abouzid K, Meng Z, Fan P, Makriyannis A. Head group analogs of arachidonylethanolamide, the endogenous cannabinoid ligand. J. Med. Chem. 1996; 39: 4515–4519.
- 18Sheskin T, Hanus L, Slager J, Vogel Z, Mechoulam R. Structural requirements for binding of anandamide-type compounds to the brain cannabinoid receptor. J. Med. Chem. 1997; 40: 659–667.
- 19Lin S, Khanolkar AD, Fan P, Goutopoulos A, Qin C, Papahadjis D, Makriyannis A. Novel analogues of arachidonylethanolamide (anandamide): affinities for the CB1 and CB2 cannabinoid receptors and metabolic stability. J. Med. Chem. 1998; 41: 5353–5361.
- 20Khanolkar AD, Makriyannis A. Structure-activity relationships of anandamide, an endogenous cannabinoid ligand. Life Sci. 1999; 65: 607–616.
- 21Thomas BF, Adams IB, Mascarella SW, Martin BR, Razdan RK. Structure-activity analysis of anandamide analogs: relationship to a cannabinoid pharmacophore. J. Med. Chem. 1996; 39: 471–479.
- 22Tong W, Collantes ER, Welsh WJ, Berglund BA, Howlett AC. Derivation of a pharmacophore model for anandamide using constrained conformational searching and comparative molecular field analysis. J. Med. Chem. 1998; 41: 4207–4215.
- 23Barnett-Norris J, Guarnieri F, Hurst DP, Reggio PH. Exploration of biologically relevant conformations of anandamide, 2-arachidonylglycerol, and their analogues using conformational memories. J. Med. Chem. 1998; 41: 4861–7482.
- 24Kent SBH, Alewood D, Alewood P, Baca M, Jones A, Schnolzer M. Total chemical synthesis of proteins: evolution of solid-phase synthetic methods illustrated by total chemical synthesis of the HIV-1 protease. In Innovations and Perspectives in Solid Phase Synthesis, R Epton (ed.). Intercept: Andover, 1992; 1–22.
- 25Zhang L, Goldammer C, Henkel B, Zühl F, Panhaus G, Jung G, Bayer E. “Magic Mixture, a powerful solvent system for solid-phase synthesis of difficult sequences”. In Innovations and Perspectives in Solid Phase Synthesis, R Epton (ed.). Mayflower Worldwide: Birmimgham, 1994; 711–716.
- 26Dauber-Osguthorpe P, Roberts VA, Osguthorpe DJ, Wolff J, Genest M, Hagler AT. Structure and energetics of ligand binding to proteins: Escherichia coli dihydrofolate reductase-trimethoprim, a drug-receptor system. Proteins: Struct., Funct., Genet. 1998; 4: 31–47.
- 27 Accelerys: San Diego, CA, 92121–3752.
- 28Frisch MJ, Trucks GW, Schlegel HB, Scuseria GE, Robb MA, Cheeseman JR, Zakrzewski VG, Montgomery JA, Stratmann RE, Burant JC, Dapprich S, Millam JM, Daniels AD, Kudin KN, Strain MC, Farkas O, Tomasi J, Barone V, Cossi M, Cammi R, Mennucci B, Pomelli C, Adamo C, Clifford S, Ochterski J, Petersson GA, Ayala PY, Cui Q, Morokuma K, Malick DK, Rabuck AD, Raghavachari K, Foresman JB, Cioslowski J, Ortiz JV, Stefanov BB, Liu G, Liashenko A, Piskorz P, Komaromi I, Gomperts R, Martin RL, Fox DJ, Keith T, Al-Laham MA, Peng CY, Nanayakkara A, Gonzalez C, Challacombe M, Gill PMV, Johnson BG, Chen W, Won MW, Andres JL, Head-Gordon M, Replogle ES, Pople JA. Gaussian 98, Revision A.6. Gaussian: Pittsburgh, PA, 1998.
- 29Ditchfield R. Self-consistent perturbation theory of diamagnetism. I. a gauge-invariant LCAO(linear combination of atomic orbitals) method for NMR chemical shifts. Mol. Phys. 1974; 27: 789–807.
- 30Rohlfing CM, Allen LC, Ditchfield R. Proton and carbon-13 chemical shifts: comparison between theory and experiment. Chem. Phys. 1984; 87: 9–15.
- 31Wolinski K, Hinton JF, Pulay P. Efficient implementation of the gauge-independent atomic orbital method for NMR chemical shift calculations. J. Am. Chem. Soc. 1990; 112: 8251–8260.
- 32Barone G, Duca D, Silvestri A, Gomez-Paloma L, Riccio R, Bifulco G. Determination of the relative stereochemistry of flexible organic compounds by ab initio methods: conformational analysis and Boltzmann averaged GIAO 13C NMR chemical shifts. Chem. Eur. J. 2002; 8(14): 3240–3245.
10.1002/1521-3765(20020715)8:14<3240::AID-CHEM3240>3.0.CO;2-G CAS PubMed Web of Science® Google Scholar
- 33Cimino P, Duca D, Gomez-Paloma L, Riccio R, Bifulco G. Comparison of different theory models and basis sets in the calculation of 13C NMR chemical shifts of natural products. Magn. Reson. Chem. 2004; 42: S26–S33.
- 34Melck D, Bisogno T, De Petrocellis L, Chuang H, Julius D, Bifulco M, Di Marzo V. Unsaturated long-chain N-acyl-vanillyl-amides (N-AVAMs): vanilloid receptor ligands that inhibit anandamide-facilitated transport and bind to CB1 cannabinoid receptors. Biochem. Biophys. Res. Commun. 1999; 262: 275–284.
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