Jennifer M. Finefield Robert M. Williams, Advisor Colorado State University December 6, 2011 Selective HDAC Inhibitor Novel Drug Delivery Method Biosynthetic Studies

Efforts Toward the Synthesis of a Selective Histone Deacetylase Inhibitor

DNA Packaging For DNA to fit within the nucleus, it must be condensed DNA is packaged into chromatin To begin packaging, DNA is wound around histones

Gene Expression: Dynamic Wrapping and Unwrapping of DNA Histone Acetyltransferase (HAT) readies DNA for transcription Histone Deacetylase (HDAC) returns DNA to the inactive state HDAC inhibitors prevent removal of acetyl residues X

Transcriptional Control HDAC inhibitors mimic the natural substrate Deacetylation is prevented, eventually leading to cell death

Zn-Dependent Histone Deacetylase Enzymes HDAC enzymes are divided into different classes Within each class, there are different isoforms Many known HDAC inhibitors display very little selectivity for class or isoform Marks, P. A., et al., Advances in Cancer Research, 2005, 137

HDAC Inhibitors and Enhancing Selectivity Many known HDAC inhibitors display very little selectivity for class or isoform Wiest, O. et al., J. Med. Chem. 2004, 47, 3409; Methot, J. L., et al., Bioorg. Med. Chem. Lett. 2008, 18, 973

Improving Selectivity of HDACi: Targeting HDAC1 and HDAC2 Moradei, O. M., et al., J. Med. Chem. 2007, 50, 5543

Improving Selectivity of HDACi: Targeting HDAC3 Finefield, J. M.; Williams, R. M.; Wiest, O.; Bradner, J. Unpublished results

Improving Selectivity of HDACi: Targeting HDAC3 Finefield, J. M.; Williams, R. M.; Wiest, O.; Bradner, J. Unpublished results Ser118, present in HDAC1/2 (green) » Tyr118 in HDAC3 (blue)

Key Disconnections

Thiazoline-Pyridine Synthesis Bowers, A., et al., Org. Lett., 2009, 11, 1301

Synthesis of the Amide Isostere Bowers, A., et al., J. Am. Chem. Soc., 2009, 131, 2900

Alternate Route

2-Thiophenyl Biaryl Synthesis

Final Steps and Future Direction ______________________________________________________________________________________________________________________________

Design and Synthesis of a Novel Drug Delivery Method Specifically Targeted to Multiple Myeloma Cells

Multiple Myeloma MM is a plasma cell malignancy that can lead to bone destruction, anaemia, hypercalcaemia, and renal insufficiency MM is associated with older age (median age 66 years) and is found to occur more often in men than women Cause of MM remains unknown Current treatments include a single high-dose of melphalan, velcade, and various combination treatments Trialx.com, Mahindra, A., et al., Blood Reviews 2010, 24, S5; Barlogie, B., et al., Blood 2004, 103, 20

Tumor Specific Oligonucleotide (MB8226) UAGGCUACGUACUUAAGCG

The Trojan Horse Nakatani, K. et al., J. Am. Chem. Soc. 2000, 122, 2172

Naphthyridine Modified MM Drugs Currently undergoing clinical trials to be used as a combination treatment for multiple myeloma Given either as a high-dose treatment or as part of a combination for the treatment of multiple myeloma

Naphthyridine Modified Vorinostat Brown, E. V., J. Org. Chem. 1965, 30, 1607; Yoshida, M. et al., Synthesis 2008, 1099; Gediya, L. K. et al. J. Med. Chem. 2005, 48, 5047

Naphthyridine Modified Vorinostat Mai, A. et al. OPPI Briefs 2001, 33, 391

Naphthyridine Modified Melphalan Nakatani, K. et al., Bioorg. Med. Chem. 2003, 11, 2347; Gullbo, J. et al., Oncol. Res. 2003, 14, 113

Preliminary Test Results

Studies on the Biosynthesis of Reverse Prenylated Indole Secondary Metabolites from Aspergillus versicolor and Aspergillus sp. MF297-2

Reverse Prenylated Indole Secondary Metabolites

Proposed Biosynthesis of the Bicyclo[2.2.2]diazaoctane Ring System Porter and Sammes Diels-Alder Proposal (1970) Enzymatic Diels-Alder Reaction Porter, A. E. A. et al., Chem. Commun. 1970, 1103; Williams, R. M., Chem. Pharm. Bull. 2002, 50, 711.

Proposed Biosynthesis of the Bicyclo[2.2.2]diazaoctane Ring System Enzyme Controlled Stereoselectivity

Reverse Prenylated Indole Secondary Metabolites

Isolation of the Notoamides: New Addition to the Stephacidin Family 2007: Aspergillus sp. MF297-2 Kato, H. et al., Angew. Chem. Int. Ed. 2007, 46, 2254

Isolation of the Notoamides: New Addition to the Stephacidin Family 2008: Aspergillus versicolor NRRL Greshock, T. J. et al., Angew. Chem. Int. Ed. 2008, 47, 3573

Antipodal Natural Products Tsukamoto, S. et al., Org. Lett. 2009, 11, 1297; Greshock, T. J. et al., Angew. Chem. Int. Ed. 2008, 47, 3573

Isolation of the Notoamides: New Addition to the Stephacidin Family : Aspergillus sp. MF297-2 Tsukamoto, S. et al: JACS, 2009, 131, 3834; JNP 2008, 71, 2064; OL 2009, 11, 1297; JNP 2010, 73, 1438

Isolation of Notoamide E: A Potential Biosynthetic Precursor : Aspergillus sp. MF297-2 Tsukamoto, S. et al., J. Am. Chem. Soc. 2009, 131, 3834

Proposed Biosynthetic Pathway: Notoamide E Greshock, T. J. et al., Angew. Chem. Int. Ed. 2008, 47, 3573

Synthesis of [ 13 C] 2 -Notoamide E Tsukamoto, S. et al., JACS 2009, 131, 3834; Grubbs, A. W. et al., TL 2005, 46, 9013; Grubbs, A. W. et al., ACIE 2007, 46, 2257

Synthesis of [ 13 C] 2 -Notoamide E Tsukamoto, S. et al., J. Am. Chem. Soc. 2009, 131, 3834

[ 13 C] 2 -Notoamide E Incorporation Study with Aspergillus sp. MF297-2 No labeled bicyclo[2.2.2]diazaoctane containing metabolites were produced Tsukamoto, S. et al., J. Am. Chem. Soc. 2009, 131, 3834

[ 13 C] 2 -Notoamide E Incorporation Study with Aspergillus versicolor Finefield, J. M.; Williams, R. M. et al., Tetrahedron Lett. 2011, 52, 1987

Possible Precursors Leading to Stephacidin A

Synthesis of Deoxybrevianamide E and 6-Hydroxydeoxybrevianamide E Kato, H.; Nakamura, Y.; Finefield, J. M.; Umaoka, H.; Nakahara, T.; Williams, R. M.; Tsukamoto, S., TL 2011, 52, 6923

Synthesis of Ketopremalbrancheamide

Biosynthetic Breakthrough: Characterization of the (  )-Notoamide Biosynthetic Gene Cluster Ding, Y.; de Wet, J. R.; Cavalcoli, J.; Li, S.; Greshock, T. J.; Miller, K. A.; Finefield, J. M.; Sunderhaus, J. D.; McAfoos, T. J.; Tsukamoto, S.; Williams, R. M.; Sherman, D. H., JACS 2010, 132, 12733

Biosynthetic Breakthrough: Identification of Two Prenyltransferases Ding, Y.; de Wet, J. R.; Cavalcoli, J.; Li, S.; Greshock, T. J.; Miller, K. A.; Finefield, J. M.; Sunderhaus, J. D.; McAfoos, T. J.; Tsukamoto, S.; Williams, R. M.; Sherman, D. H., JACS 2010, 132, 12733

Early Steps in the Biosynthetic Pathway

Feeding Study with [ 13 C] 2 -[ 15 N]-6- Hydroxydeoxybrevianamide E No incorporation into advanced metabolites Finefield, J. M.; Williams, R. M. et al., JOC 2011, 76, 5954; Finefield, J. M.; Williams, R. M.; Tsukamoto, S. et al., TL 2011, 52, 6923

Possible Enantio-diverging Pathways from Notoamide S

Notoamide S Incorporation Study with Aspergillus versicolor Unlabeled synthesis of notoamide S: McAfoos, T. J. et al., Heterocycles 2010, 82, 461 Results from feeding study: Finefield, J. M.; Tsukamoto, S.; Williams, R. M. et al., unpublished results

Notoamide S Incorporation Study with Aspergillus sp. MF297-2 Tsukamoto, S. et al., unpublished results

Notoamide S: Additional Biosynthetic Insight

Notoamide T: Precursor Incorporation Studies ________________________________________________________________________________________________________________________________ Finefield, J. M., Tsukamoto, S.; Williams, R. M. et al., unpublished results

Notoamide S: Additional Biosynthetic Insight

Biosynthetic Precursor Incorporation Study ________________________________________________________________________________________________________________________________ Finefield, J. M.; Tsukamoto, S.; Williams, R. M. et al., Org. Lett. 2011, 13, 3802

Characterization of the (  )-Notoamide Biosynthetic Gene Cluster (  )-Notoamide Biosynthetic Gene Cluster (+)-Notoamide Biosynthetic Gene Cluster Li, S.; Sherman, D. H. et al., unpublished results

Current Postulated Biogenesis of the Notoamides and Stephacidins

Summary

Acknowledgements Prof. Robert M. Williams Williams Research Group Prof. David H. Sherman Sherman Research Group Prof. Sachiko Tsukamoto Tsukamoto Research Group Dr. James Berenson