Fiorillo vi ewd_sy_siena_2016
•Download as PPTX, PDF•
1 like•279 views
VI EWDSy: European Workshop in Drug Synthesis Certosa di Pontignano, Siena, Italy, 15-19 May 2016
Recommended
Recommended
More Related Content
What's hot
What's hot (20)
Similar to Fiorillo vi ewd_sy_siena_2016
Similar to Fiorillo vi ewd_sy_siena_2016 (20)
Recently uploaded
Recently uploaded (20)
Fiorillo vi ewd_sy_siena_2016
- 1. Recent advances in antitumour berberine Gaetano Fiorillo, Tanjia Monir Syeda , Paolo Lombardi via G. Di Vittorio 70, 20026 Novate Milanese, Milano, Italy Email: staff@naxospharma.eu Siena, 15-19 May 2016
- 2. Berberine Extracted from plants of the genus Berberis, Coptis and others. In use in the Ayurvedic and Chinese medicines. Anti-microbial/parasitic, Anti-diarrheal, anti-inflammatory, Anti-arryhthmic, Cholesterol-lowering Anticancer1,2 Berberine Chloride 1L. M. Guamán Ortiz, P.Lombardi, M. Tillhon, A. I. Scovassi, Molecules ,2014, 19, 12349-12367 2 L. M. Guamán Ortiz, P.Lombardi, M. Tillhon, A. I. Scovassi, Biochemical Pharmacology, 2012, 84, 1260–1267 2011: 7 clinical trials 2016: 25 clinical trials
- 3. Mondello, R. et al.,Bioorg Med Chem, 2003, 505–514 (NMR Studies) Gratteri, P et al.Chem. Commun. 2011, 4917- 4919 (RX studies) minor groove binding Berberine DNA Interaction Mechanism “ Interaction between nucleic acids and berberine sulfate “ Journal of Cellular Biology, 15, 1962, 589 …has been reported since decades… Minor groove binding or Intercalation ????
- 4. Berberine represents an interesting and attractive natural lead compound Chemical modifications might select more specific medical indications resulting in derivatives with better (or different) biological effects compared to the parent berberine Performing rational chemical modifications of berberine structure led to a new class of derivatives with antitumour properties Chemical Programme Berberine
- 5. Aromatic interactions are ubiquitous in nature, their geometry is relevant for the molecular recognition in biological systems 1 Chemical Programme L = Linker with different functionalities 1 Waters ML, Curr Opin Chem Biol. 2002, 6, 736
- 6. from very low to low yields - better with activated halides or iodides - berberine back from loss of acetone major by-product Alkylation of enamine (7,8-dihydroberberine) Berberine derivatives Synthetic Methods_1 1) from low to moderate yields - berberine and tetrahydroberberine from disproportionation of enamine as major by-products 2)
- 7. generally from good to very good yields Uncommon aldehyde-enamine condensation1,2 1 Cook, AG, Enamines Synthesis, Structure and reaction, 1988, 200 2 Iwasa, K, et al., Planta Medica, 1997, 196 Berberine derivatives Synthetic Methods_2
- 8. 1) Commercially available aldehyde 2) Commercially available alcohol followed by oxidation (e.g. : PCC or TEMPO) 3) Homologation starting from the above Berberine Derivatives Aldehyde Intermediates_1 Commercially available
- 10. Improved DNA intereaction of berberine derivatives_1 1.77 0.35 2.11 11.01 7.6 7.58 6.8 0 2 4 6 8 10 12 Kix10-5(M-1) Interaction costants of NAXs 1 1.77 0.48 0.51 7.07 10.04 8.90 7.48 0 2 4 6 8 10 12 Kix10-5(M-1) Interaction costants of NAXs2 1D. Bhowmik, M. Hossain, F. Buzzetti, R. D’Auria, P. Lombardi, G.S.Kumar, J. Phys. Chem. B, 2012, 116, 2314−24. 2D. Bhowmik, F. Buzzetti, G. Fiorillo, F. Orzi, T. Syeda Monir, P. Lombardi, G.S. Kumar, Med. Chem. Comm., 2014, 5, 226-31. n = 3 n = 4 Monophenyl Derivatives Diphenyl Derivatives
- 11. S. Chatterjee, S. Mallick, F. Buzzetti, G. Fiorillo, T. M. Syeda, Paolo Lombardi, K. Das Saha, G. S. Kumar, RCS Adv., 2015, 5, 90632 n = 5 0 2 4 6 8 10 12 Kix10-5(M-1) Interaction costants of NAXs Pyridylalkyl Derivatives Improved DNA intereaction of berberine derivatives_2
- 12. Binding to human telomeric G quadruplex P. Gratteri, M. Ferraroni, C. Bazzicalupi, F. Papi, G. Fiorillo, L. M. Guamán Ortiz, A. Nocentini, A. I. Scovassi, P. Lombardi Chemistry An Asian Journal, 2016, 11(7),1107-15 NAX 053 – d[TAG3(T2AG3)T] adduct crystal structure
- 13. Our1 and others’ studies identified berberine as a novel, non specific inhibitor of the nascent synthesis of some proteins, supposedly acting as a RNA silencing agent Berberine effect on TS expression 1G. Marverti, A. Ligabue, P. Lombardi, S. Ferrari, M. G. Monti, C. Frassinetti, M.P. Costi, Int. Journal of Oncology, 2013, 43, 129 2008 cells = cisplatin sensitive C13 cells = cisplatin resistant
- 14. Berberine NAX035NAX012 Berberine derivatives: effect on TS over-expression in mesothelioma cell lines TS levels: time-course in mesothelioma MSTO-211H cells at the IC50 dose at 72 h NAX038
- 15. Berberine Derivatives Antiproliferactive effects against human mesothelioma cell lines STO, MESOII = peritoneal mesothelima cell lines MSTO = pleural mesothelioma cell lines 0 1 2 3 4 5 6 7 8 9 IC50[mM] Mesothelioma cell lines STO MESOII MSTO 0 1 2 3 4 5 6 7 8 9 IC50[mM] Mesothelioma cell lines STO MESOII MSTO
- 16. Antitumour activity of i.p. and oral NAX035, on the peritoneal STO human mesothelioma s.c. xenografted in nude mice Route Dose mg/kg TVI% (+32) (PvsControls) Max BWL% TOX i.p. 1 52 (0.1181) 8 0/9 p.o. 10 72 (0.0434) 10 0/9 p.o. 15 74 (0.0373) 5 0/8 Correlation between TS protein levels in vitro and in vivo in tumour tissue samples examined at the end of the p.o. treatment period
- 17. NAX 012 NAX 013 NAX 014 NAX 035 Berberine 24 h 94.2±1.2 >100 52.3±3.2 >100 91.8±2.8 48 h 46.6±2.5 >100 30.7±2.1 >100 58.4±1.9 72 h 31.9±2.9 >100 26.5±6.7 48.6±6.7 36.0±1.8 Antiproliferative effect (IC50 mM) Method - Alamar Blue assay. The number of viable cells after treatment is expressed as a % of the vehicle treated control M. Provinciali, P. Lombardi, et al, Biofactors, 39, 2013, 672-679 Berberine derivatives Antiproliferactive effects on HER2+ human Breast Cancer cells (SK-BR-3) (SK-BR-3)
- 18. Tumour Number Tumour Growth Inhibition High % Tumour Free Mice NAX014: antitumour efficacy in HER-2/neu spontaneous tumours transgenic female mice Both i.p. [2.5mg/kg-(2xweek)x12] and oral [20mg/kg-(2xweek)x8] administrations of NAX014 reduced tumour volume and number, delayed the onset and progression of HER2+ BC compared to control group Tumour Growth Inhibition Tumour Number i.p. p. o. Provinciali, M., Lombardi, P. et al., Carcinogenesis, 2015, 1–11
- 19. HER2 p-HER2 β-actin Ctrl Different NAX compounds specifically inhibit the expression of different proteins Lapatinib+ Trastuzumab NAX012NAX014 NAX035
- 20. Innovative proprietary compound, structurally related to the plant isoquinoline alkaloid berberine. Novel mechanism of action, targeting the expression of TS protein, differently from previous TS inhibitors Efficacy on chemoresistant tumour cells Antitumour efficacy and tolerability at the effective doses by oral and i.p. administration in a human mesothelioma xenografted nude mice Conclusions: NAX 035
- 21. Conclusions: NAX 014 anticancer and anti-metastatic efficacy on HER2+ tumours in vitro activity at µM concentrations in vivo tolerability by i.p.and oral administration at the effective dose Innovative proprietary compound, structurally related to the plant isoquinoline alkaloid berberine Unique ability to reduce cellular HER2 expression via a postulated novel mechanism
- 22. Financial supports were provided by Ministero dello Sviluppo Economico (Grant 01705) to Naxospharma srl (Project coordinator) and to Istituto Tumori Milano and by Agència per a la competitivitat de l'empresa ACC1O (Grant RDNET11-1-0001) to Aromics SL, Barcelona under the 6th call of the EuroTransBio initiative, Transnational joint project BERTA (BERberine as antiTumour Agents). Aknowledgeme nts Regione Lombardia (Grant 13810040) to Naxospharma srl and to Istituto di Genetica Molecolare – CNR Pavia, Project PLANT CELL Ente Cassa di Risparmio di Firenze (Grant 2014.0309) to Department NEUROFARBA, University of Firenze
Editor's Notes
- I’d like to thank the scientific committee for the opportunity to present our research work
- Berberine is an isoquinoline quaternary alkaloid extracted of from plants of the genus Berberis and Coptis. It shows diverse pharmacological activities: Anti-microbial/parasitic, Anti-diarrheal, anti-inflammatory, Anti-arryhthmic, Cholesterol-lowering, and anticancer We have written two reviews in collaboration with CNR of Pavia, one of which specifically addresses anticancer properties of berberine In two thousand eleven there were 7 ongoing clinical trials, which increased to 25 today
- Now, how berberine explicates its anticancer effects? Evita di dire ciò, non si può dimostralo con certezza Interaction between nucleic acids and berberine has been reported since nineteen sixty two. The question whether the alkaoild is a minor groove binder or an intercalator was matter of debate
- In any case… read…
- Our chemistry programme was directed to the synthesis of novel Berberine derivatives characterized by the presence of (hetero)aromatic groups bonded to the 13-position of the parent alkaloid skeleton through a linker of variable length and functionality. That would possibly creates a geometric propensity for additional stacking-type, non-covalent aromatic interactions with respect to the parent Berberine. Aromatic interactions are ubiquitous in nature, their geometry is relevant for the molecular recognition in biological systems That could result in better (or different) biological effects, possibly ameliorated antitumour properties. Here on the right are some examplary derivatives
- Synthetic methods of the prior art for C-13 functionalization imply: preparation of a reactive acetonylberberine intermediate, followed by enamine alkyl halide alkylation and concomitant expulsion of acetone to restore the isoquinoline quaternary salt of the desired derivative reduction of berberine to dihiydro berberine, followed by enamine alkyl alide alkylation to obtain an instable iminium ion, reduction with sodium borohydride, oxidation with appropriate reagents to obtain the isoquinoline quaternary salt of the desired derivative
- In our hands, the most viable and direct preparative method was based on this uncommon enamine-aldehyde condensation. The postulated mechanism implies an unstable alcoholic intermediate wich looses water and H+ to obtain the desired isoquinoline quaternary salt derivative
- Good, we use aldehyde intermediates when commercially available, or commercially available alcohol followed by oxidation or we use homologation procedure. For example starting from diphenyl propanol we obtain diphenyl propanal with tempo or pcc oxidation. Than, aldehyde elongation by wittig reaction followed methanolysis or hydrolysis.
- Moremore, wittig reaction without solvent, hydrogenation, reduction and tempo oxidation to obtain pyridyl alkyl derivative. Alternatively, we use Darzen’s reaction as a homologation procedure.
- This slide shows the partnership with Indian institute of chemical biology to estabilish the interaction costants of some of our berberine derivatives with calf thymus DNA Click With respect to parent berberine, the interaction depends on the length of the linker For the monophenylalkyl derivatives the maximum interaction is obtained when the linker has three carbon atoms whereas for the diphenylalkyl the maximum interaction is obtained with four carbon atoms
- Similarly for pyridyl alkyl derivatives, where the best constant interaction is with five carbon atoms linker. We did not investigate longer linkers.
- Crystal structure of the adduct formed by NAX053 and the human telomeric sequence. The interaction is a Pi stacking. click This compound affects the viability of HeLa cancer cell lines in a dose-dependent manner.
- Our1 and others’ studies identified berberine as a novel, non specific inhibitor of the nascent synthesis of some proteins, supposedly acting as a RNA silencing agent. We found that berberine inhibits the synthesis of thymidylate synthase (an imporant protein overexpressed in many cancers). Noteworthy, berberine was more effective in cisplatin resistant cancer cells than in the sensitive ones.
- Some of our berberine derivatives show a more pronounced effect on TS expression of, for example, pleural mesothelioma MSTO cell lines with respect to berberine
- This slide shows the antiproliferative activity against one pleural and two peritoneal mesotelhelioma cell lines, for the monophenyl and diphenyl classes, resepctively. Non ti perdere qui a fare commenti tra attività e struttura.
- This in vivo study refers to the efficacy of NAX035 (administered either intraperitoneally or orally) in a peritoneal STO human mesothelioma s.c. xenografted in nude mice. TVI = tumour volume inhibition BWL% = percent body wight loss TOX = 0 dead animals in the treated group ( 9 or 8) Note correlation between downregulation of TS protein in vitro correlates with downregulation of TS protein in tumour tissue of treated animal = in vivo target modulation
- We investigated also the antiproliferative activity of some of our novel berberine derivatives on a HER2+ human Breast Cancer cell line, in collaboration with IRCCS-INRCA in Ancona Also on that basis of other pharmacological investigations (not reported here) NAX014 was chosen for in vivo studies
- This in vivo study refers to a model of spontaneous development of HER2 positive breast cancer in transgenic mice, very similar to what happens in women. Note that time course scale is in WEEKS Quindi leggi Both ip and...
- The antitumour efficacy of NAX014 is due to the inhibition of the expression of HER2 protein, whereas current anti breast cancer drugs transtuzumab (antibody) and lapatinib (kinase inhibitor) exert different mechanims BUT DO NOT AFFECT THE LEVEL OF THE PROTEIN, WHICH REMAINS OVEREXPRESSED Noteworthy, NAX compounds which down regulate TS (e.g. NAX012 and NAX035) do not affect HER2 expression, and NAX014 which downregulates HER2, does not affect TS expression.
- In conlusion we have selected two lead compound, The first is nax thirty five (e poi leggo)
- And nax 014 that have unique ability to reduce ccellular HER2 expression via a postulated novel mechanism
- Finally i want to thank the the funding agencies which supported our and our partners’ research