- Email: [email protected]
Free radical-scavenging activity of olive oil phenolics
S26
101
103
FREE RADICAL-SCAVENGING ACTIVITY OF OLIVE OIL PHENOLICS Francesco Visioli and Claudio Galli lnstltute of Pharmacological Sciences, Via Balzaretti 9, 20133 Milan, Italy. [email protected]
ANTIOXIDANT PROPERTIES OF GRAPE PROCYANIDINS . . mFpellloo, M. Cnrini”, G. AIdiS’, F. B&i*, G. Rossoni’, E. Bombardelli*and P. Morazmai’ “Istiluto Cbimicn Fwmacwtlco Tossicologico, V.le Abmzi 42, 20131 Milan, Italy; ‘Dipartimento di Farmacologia, Chemioterapia e Tossicologia Medica, Via Vanvitelli 32, 20129 Milan, Italy; ‘Indena S.p.A., V.le Ortles 12,20139 Milan, Italy. The antioxidant properties of procyanidins from Viris vini&u seeds has been extensively investigated in different m models, cell-free, membranous (phosphatidylcholine liposomes) and cell systems (crythrocytes, leukocytes). Pwyanidins behave as potent preventive and chain-breaking antioxidants, vitamin E recycling agents, iron/copper chelators and inhibitors of the respiratory burst of human activated neutrophils. In post-ischemic reperfiwd rabbit heart Langerdorff preparations they exert strong antiarrhythmic and cardioprotective effects due to the entrapping of reactive oxygen species. The antioxidant potential and the protective effects on cardiac mechanics following ischemia/reperfusion damage were also demonstrated in it&w studies (3 weeks supplementation of young and aged rats with procyanidins complexed with soybean lecithin, Leucoselccty. Procyanidins markedly increase the plasma antioxidant defenses of both young and aged rats (TRAP value and ascorbic acid); in parallel, a highly significant recovery in the mechanical performances of the hearts from stunning/reperfusion was found, determined by measuring left ventricular developed pressure, coronary perfusion pressure and the release of creatine kinase and &keto-PGF,, in the pe&asate. A similar increase in total
such pro erties may be relevant to the observed protection from C l! D and cancer. Finally, since a considerable amount of such corn ounds is currently discarded as olive mill waste water, t Re recovery of the latter may bear interesting applications in preservative chemistty.
procyanidins-@sated hypercholesterolemic
102
104
Methoxylated analoguer of cocalne: ldentltication of a weak ‘antagonlst”. w. Robert[*, E. Andrisano? D. Simonib,A.P. Kozikowski
IDENTIFICATION
aDipartimento Salmeloro 6, bDipartimento cGeorgetown
dl Scienze Farmaceutiche. LJnivesit(l di Bologna, Via 40126 Bologna, Italy. di Scienze Famraceutiche, Universita di Ferrara, Italy. University , Washington DC.
Our own efforts to acquire a better understanding of the topographyof the cocainereceptorwith the aim toward identifyng a possiblecocaine antagonist, led us to investlgate modifications to
various region of this molecule. Recently, we +potied that racemic two-carbon bridae methoxviated cocaine were found to antaaonize. albait weakly,cocalne’s abi%ty to inhibit dopamlne reuptake. iereln, we now report a novel enantioselectlve approach for the preparation of two-carbon bridge methoxylated cocaine which In turn allows, concomitant, facile attribution of their absolute configuration. All new synthesized analogues were examined for their ability to diaolacel3Hl mazindol blndina and to Inhibit hiah-affinltv uotake of [3i] dopamina into striatal nerve andings (synaptoiomae). Of interest. adiscreoancie wasobserved in the blndina and dooamine uptake inhibition’ data. For the pseudococaine-like-derivatives it is assessed the idea of pursuing a cocaine -based approach for the development of cocaine antagonist.
(-)
cocrlno
6- and 7.methoxylatrd and pseudo Cocalnas
cocalnoa
0. Simoni, J Stoelwinder, A.P. Kozikowski, K.M. Johnson, Bergmann, R.G. Ball. J. Med. Chem., 1993, 36. 3975.
plasma antioxidant capacity and ascorbate levels was observed in healthy volunteers after 5 days procyanidins supplementation. Finally a marked prevention in the formation of atherosclerotic lesions was observed in rabbits.
POTENT PEIARMACODYNAMIC OF PROPERTIES OF BOSWELLIC ACIDS T. Svrovcts, B. Btichelc,K. Belancr, W. Sauter and Th. Simmet Institute of Pharmacology, Toxicology and Natural products, Hehnholkstr. 20, D-89081 Uhn, Germany
Boswcllic acids, which arc pentacyclic triterpenes, were extracted from the gum resin of Bow&a carfwi Birdw., purified by HPLC and identified by mass spechometry and nuclear magnetic resouancc spectroscopy. purity was greater than 99%. Ace@-P-boswellic acid as well as acetyl-1 I-keto-P-boswellic acid exerted concentration-dependent cyctotoxic effects in hvo established malignant human glioma cell lines, U-87 MG and U-373 MG. Cytotoxicity was no1 due to apoptosis as shown by a lack of morphological signs of apoptosis or DNA laddering. Morcovcr, neither the TUXEL method nor the histone assay provided any evidence for apoptoris. Tubulin was not inhibited either, but distinct morphological changes of the nuclei stained with Hocchst 33258 hinted to a potential inhibition of topoisomcrases. Indeed, nuclear extracts from both cell lines used exhibited high activities of topoisomeraae I and II activity as shown by relaxation of plasmid pBR322 and dccatation of kinctoplast DNA, respectively. Further experiments were performed with purified human topoisomezase I and iopoiaomeraae Ilp170 isofonn. Effects of boswellic acids on kqoisomerase activity were tested in the so-called relaxation assays using pBR322 and pRYG plasmid substrate DNA. Acctyl-P-boswellic acid as well BP ace@-1 I-keto-P-boswellic acid were found to be potent inhibitors of human topoisomerasc I and Il activity, specifically, when compared 1o ihe standard inhibitors camptotbccin and amaacrine, respectively. Indeed, the inhibitory potency of the boswellic acids mentioned exceeded that of the standard inhibitors. Among various other pentacyclic triterpenes, only brmlinic acid and oleanolic acid exhibited weak inhibitory activity at high concentrationa such as
100 pM. Various experimental approaches show that boswcllic acids do not inter&ate into DNA. Moreover, in contrast to most topoisomcrllsc inhibitors, they do not stabilize rhe cleavable complex, but inhibit topoisomcrasc activity
J. S.
apparently on the enzyme level. Baswellic acids represent a novel greup of topoisomerase inhibitors with a distinct mechanism of action. Due to their hpophilicity, they nught be particularly useful1 for the chemotherapeutic treatmentof humanmalignantbrain tumors.
101
103
FREE RADICAL-SCAVENGING ACTIVITY OF OLIVE OIL PHENOLICS Francesco Visioli and Claudio Galli lnstltute of Pharmacological Sciences, Via Balzaretti 9, 20133 Milan, Italy. [email protected]
ANTIOXIDANT PROPERTIES OF GRAPE PROCYANIDINS . . mFpellloo, M. Cnrini”, G. AIdiS’, F. B&i*, G. Rossoni’, E. Bombardelli*and P. Morazmai’ “Istiluto Cbimicn Fwmacwtlco Tossicologico, V.le Abmzi 42, 20131 Milan, Italy; ‘Dipartimento di Farmacologia, Chemioterapia e Tossicologia Medica, Via Vanvitelli 32, 20129 Milan, Italy; ‘Indena S.p.A., V.le Ortles 12,20139 Milan, Italy. The antioxidant properties of procyanidins from Viris vini&u seeds has been extensively investigated in different m models, cell-free, membranous (phosphatidylcholine liposomes) and cell systems (crythrocytes, leukocytes). Pwyanidins behave as potent preventive and chain-breaking antioxidants, vitamin E recycling agents, iron/copper chelators and inhibitors of the respiratory burst of human activated neutrophils. In post-ischemic reperfiwd rabbit heart Langerdorff preparations they exert strong antiarrhythmic and cardioprotective effects due to the entrapping of reactive oxygen species. The antioxidant potential and the protective effects on cardiac mechanics following ischemia/reperfusion damage were also demonstrated in it&w studies (3 weeks supplementation of young and aged rats with procyanidins complexed with soybean lecithin, Leucoselccty. Procyanidins markedly increase the plasma antioxidant defenses of both young and aged rats (TRAP value and ascorbic acid); in parallel, a highly significant recovery in the mechanical performances of the hearts from stunning/reperfusion was found, determined by measuring left ventricular developed pressure, coronary perfusion pressure and the release of creatine kinase and &keto-PGF,, in the pe&asate. A similar increase in total
such pro erties may be relevant to the observed protection from C l! D and cancer. Finally, since a considerable amount of such corn ounds is currently discarded as olive mill waste water, t Re recovery of the latter may bear interesting applications in preservative chemistty.
procyanidins-@sated hypercholesterolemic
102
104
Methoxylated analoguer of cocalne: ldentltication of a weak ‘antagonlst”. w. Robert[*, E. Andrisano? D. Simonib,A.P. Kozikowski
IDENTIFICATION
aDipartimento Salmeloro 6, bDipartimento cGeorgetown
dl Scienze Farmaceutiche. LJnivesit(l di Bologna, Via 40126 Bologna, Italy. di Scienze Famraceutiche, Universita di Ferrara, Italy. University , Washington DC.
Our own efforts to acquire a better understanding of the topographyof the cocainereceptorwith the aim toward identifyng a possiblecocaine antagonist, led us to investlgate modifications to
various region of this molecule. Recently, we +potied that racemic two-carbon bridae methoxviated cocaine were found to antaaonize. albait weakly,cocalne’s abi%ty to inhibit dopamlne reuptake. iereln, we now report a novel enantioselectlve approach for the preparation of two-carbon bridge methoxylated cocaine which In turn allows, concomitant, facile attribution of their absolute configuration. All new synthesized analogues were examined for their ability to diaolacel3Hl mazindol blndina and to Inhibit hiah-affinltv uotake of [3i] dopamina into striatal nerve andings (synaptoiomae). Of interest. adiscreoancie wasobserved in the blndina and dooamine uptake inhibition’ data. For the pseudococaine-like-derivatives it is assessed the idea of pursuing a cocaine -based approach for the development of cocaine antagonist.
(-)
cocrlno
6- and 7.methoxylatrd and pseudo Cocalnas
cocalnoa
0. Simoni, J Stoelwinder, A.P. Kozikowski, K.M. Johnson, Bergmann, R.G. Ball. J. Med. Chem., 1993, 36. 3975.
plasma antioxidant capacity and ascorbate levels was observed in healthy volunteers after 5 days procyanidins supplementation. Finally a marked prevention in the formation of atherosclerotic lesions was observed in rabbits.
POTENT PEIARMACODYNAMIC OF PROPERTIES OF BOSWELLIC ACIDS T. Svrovcts, B. Btichelc,K. Belancr, W. Sauter and Th. Simmet Institute of Pharmacology, Toxicology and Natural products, Hehnholkstr. 20, D-89081 Uhn, Germany
Boswcllic acids, which arc pentacyclic triterpenes, were extracted from the gum resin of Bow&a carfwi Birdw., purified by HPLC and identified by mass spechometry and nuclear magnetic resouancc spectroscopy. purity was greater than 99%. Ace@-P-boswellic acid as well as acetyl-1 I-keto-P-boswellic acid exerted concentration-dependent cyctotoxic effects in hvo established malignant human glioma cell lines, U-87 MG and U-373 MG. Cytotoxicity was no1 due to apoptosis as shown by a lack of morphological signs of apoptosis or DNA laddering. Morcovcr, neither the TUXEL method nor the histone assay provided any evidence for apoptoris. Tubulin was not inhibited either, but distinct morphological changes of the nuclei stained with Hocchst 33258 hinted to a potential inhibition of topoisomcrases. Indeed, nuclear extracts from both cell lines used exhibited high activities of topoisomeraae I and II activity as shown by relaxation of plasmid pBR322 and dccatation of kinctoplast DNA, respectively. Further experiments were performed with purified human topoisomezase I and iopoiaomeraae Ilp170 isofonn. Effects of boswellic acids on kqoisomerase activity were tested in the so-called relaxation assays using pBR322 and pRYG plasmid substrate DNA. Acctyl-P-boswellic acid as well BP ace@-1 I-keto-P-boswellic acid were found to be potent inhibitors of human topoisomerasc I and Il activity, specifically, when compared 1o ihe standard inhibitors camptotbccin and amaacrine, respectively. Indeed, the inhibitory potency of the boswellic acids mentioned exceeded that of the standard inhibitors. Among various other pentacyclic triterpenes, only brmlinic acid and oleanolic acid exhibited weak inhibitory activity at high concentrationa such as
100 pM. Various experimental approaches show that boswcllic acids do not inter&ate into DNA. Moreover, in contrast to most topoisomcrllsc inhibitors, they do not stabilize rhe cleavable complex, but inhibit topoisomcrasc activity
J. S.
apparently on the enzyme level. Baswellic acids represent a novel greup of topoisomerase inhibitors with a distinct mechanism of action. Due to their hpophilicity, they nught be particularly useful1 for the chemotherapeutic treatmentof humanmalignantbrain tumors.