浜松工業会東京支部若手会員活性化講演会（平成26年7月23日）工学部からの創薬研究 “似て非なるもの”を活用するものづくり静岡大学大学院・工学研究科鳴海哲夫似て非なるもの “見かけは似ているが、本質的には違うもの” コカ・コーラ 45 kcal/100 mL 甘味料：砂糖（スクロース）コカ・コーラゼロ 0.5 kcal/100 mL 甘味料：アスパルテーム異なる分子構造を有しながら、生体が同様に「甘味」を感じる“似て非なる”分子アミド結合１）タンパク質やペプチドの主鎖骨格を形成する普遍的な共通構造２）水素結合能により、二次構造や高次構造の形成に寄与３）生理活性分子や医薬品の重要な構造 →医薬品上位200化合物の内、95化合物がアミド結合を含むアミド結合の特性に基づいて新たな機能性分子の創製や効率的合成法の開発 Concept: Replacement of Amide and Alkene O R R1 N H amide X Structural isosterism R1 R H alkene Hann, M.M. et al., J. Chem. Soc., Perkin Trans. 1, 1982, 307. H N R2 O R1 N H O Amide to Alkene Cell Biology R1 R2 H Chemical Biology O Alkene-type isosteres Native peptide Chemical Biology X H N Isostere-based Design X = H, Me, F, Cl, CF3 Alkene to Amide Organic Chemistry Small change, but still great advantages! Medicinal Chemistry Native Peptide and its Alkene Isosteres Xaa N H O H N O Xaa N H Yaa O O H N N H Yaa X • Ground-State Amide Equivalents O H Xaa • Peptidomimetics for Medicinal Chemistry Yaa Alkene-Type Isosteres • Chemical Probes for Native Peptides Biochemistry Class I Xaa N H H H Class II O N H Yaa (E)-Alkene Dipeptide Isostere (EADI) Class III N H Yaa (Z)-F-alkene Dipeptide Isostere (FADI) Xaa N H F O Me Yaa (E)-Me-alkene Dipeptide Isostere (FADI) O Xaa O Xaa Yaa (Z)-Cl-alkene Dipeptide Isostere (CADI) O Xaa N H Cl CF3 Yaa (Z)-CF 3-alkene Dipeptide Isostere (FADI) 「アルケン型ジペプチドイソスターの効率的合成法の開発と生理活性ペプチドの構造活性相関研究への応用」有機合成化学協会誌、2008年、66巻、846-857. Biolgical Evaluation of Alkene Pseudopeptides Org. Biomol. Chem., 2010, 8, 616. Biolgical Evaluation of Alkene Pseudopeptides Biopolymers 2007, 88, 272. Entry X 1 Activity (%)a EC50 (nM) 96.5 ± 0.3 3.1 2 (transoid) 75.8 ± 1.6 7.6 3 (cisoid) 2.0 ± 0.3 — 4 (transoid) 2.6 ± 0.1 — 5 (cisoid) 5.4 ± 0.1 — aActivity is based on the relative maximum agonistic activity at 10 nM of a compound to 1 mM kisspeptin-10 (%). 100% = maximum agonistic activity signal at 1 mM kisspeptin-10. Ongoing and Future Study “Caged” Compounds in Chemical Biology How can we control biological activities (spatially and temporally)? Addition of bioactive compound Responses to stimulation 光感受性保護基光 hn ² ケージド化合物 "IN-ACTIVE" "ACTIVE" 生理活性分子細胞生命現象のリアルタイム解析により、創薬標的となる過渡的複合体の機能解明 Coumarin-4-ylmethyl Groups Large molar absorption coefficients at longer wavelength Fast release rates with improved stability Two-photon photolysis with practically useful absorption cross sections (720-900 nm) Recent Review of Courmain-type PPGs: Givens, et al. Photochem. Photobiol. Sci. 2012, 11, 472. Caged Ligands for Photoactivation of Protein Kinase C “IN-ACTIVE” 0 min “ACTIVE” 10 min Nomura, W., Narumi, T., et al., ChemBioChem, 2011, 12, 535-539. Translocation of Activated PKCδ within the Golgi-stained cells HeLa cells 0 min 10 min EGFP-PKCδ BODIPY TR ceramide merged image Photoirradiation induced translocation of PKCδ to Golgi apparatus. Slow response against photoirradiation Wataru Nomura, Yuki Serizawa Concept: Isosteric Replacement of Alkene and Amide O R X Structural isosterism R1 N H amide R1 R H alkene Hann, M.M. et al., J. Chem. Soc., Perkin Trans. 1, 1982, 307. Br HO Bhc enol double bond Br Isostere-based design O O alkene-to-amide replacement HO aromatization Br O N O H amide bond N O 8-aza-Bhc O O Hydrophilic Properties of 8-Azacoumarin and Bhc Derivatives 2.5 mM in PBS CS: 602 mM pKa: 5.88 ✓ Inductive effect CS: 10832 mM pKa: 4.22 2.5 mM in PBS ✓ Strong inductive effect ✓ H-bonding interaction Application of 8-aza-Bhc groups to Bioactive Compounds X X Br Br Isostere-based design HO O Excellent water solubility HO O 6-Bhc Excellent photoreactivity N O O 8-Aza-3-Bhc Efficient photorelease at lower pH Novel caging group HO O O O 3 O O NH 2 Br HO estertype H 2N OH O HN carbamatetype N O carbonatetype UV, 365 nm, 25 s "85% conv." O O O O Br Br O HO N O 3 HO N N N O O O HO UV, 365 nm, 20 s "90% conv." O 6 N O O UV, 365 nm, 80 s "84% conv." Promising applications to alcohol, amine and carbonyl functional groups N HIV-1外被タンパク質gp120の構造変化誘起を指向した低分子CD4ミミックの構造活性相関研究 Viral Attachment CD4 gp120 Coreceptor Binding Virus-Cell Fusion Entry HIV-1 Conformational Change of gp120 CD4 mimic Phe43 Cavity (CD4 Binding Site) Coreceptor Binding Site (X = Cl, Br, Me) SAR Study anti-HIV activity, cytotoxicity CD4 mimicry on conformational change Narumi, Narumi, Narumi, Narumi, T., et al., BMCL 2010, 20, 354-358. T., et al., BMCL 2010, 20, 5853-5858. T., et al., BMC 2011, 19, 6735-6742. T., et al., BMC 2013, 21, 2518-2526. 中和抗体活性増進剤: エンベローププロテインオープナー NAb +NBD +sCD4 anti V3+NBD CD4ミミック誘導体はgp120 の構造変化を誘起することで、中和抗体活性を増進させる。 CD4i+NBD CD4i anti V3 Yoshimura K, Harada S, Tamamura H, Matsushita S, et al., J.Virol., 2010 Our SAR Studies BMCL 2010, 20, 354. BMCL 2010, 20, 5853. BMC 2011, 19, 6735. BMC 2013, 21, 2518 Hiroshi Arai 抗HIV活性が約４０倍向上した誘導体(HAR-431)の創出に成功 2013年１０月１日から静岡大学工学部で、鳴海研究室をはじめました。今後とも手厚い（手荒い？）ご指導よろしくお願いします。創薬を目指して新しい有機分子を作り生命現象を科学する Research Group 大学院生、研究生、共同研究を募集しています。鳴海哲夫連絡先：[email protected]