변형뉴클레오티드 합성 및 이를 이용한 mRNA번역 저해 효율 연구 = Synthesis of Modified Nucleotides and a Study on Their mRNA Translation Inhibition Efficiency

다국어 초록 (Multilingual Abstract)

Eukaryotic translation initiation is critically governed by recognition of the mRNA 5′ cap by eukaryotic translation initiation factor 4E (eIF4E), a central component of the eIF4F complex. Because dysregulated cap-dependent translation and elevated eIF4E activity are implicated in multiple diseases, including cancer, competitive modulation of eIF4E–cap interactions represents an attractive strategy for both mechanistic studies and therapeutic exploration. Here, we designed and synthesized a series of N²- and N⁷-modified guanosine derivatives intended to competitively engage the eIF4E cap-binding site, and systematically evaluated their binding and translation inhibition activities in a cell-free translation system. We first generated a set of guanosine monophosphate (GMP) mononucleotide analogues using a simple and scalable synthetic route and quantified their inhibitory potency relative to the reference compound m⁷GTP. Several mononucleotide derivatives exhibited enhanced translation inhibition compared with m⁷GTP, demonstrating that appropriate substituent installation can substantially improve competitive activity even within a mononucleotide scaffold. To further validate a cap-analogue design strategy, we synthesized corresponding dinucleotide analogues based on selected mononucleotide leads. The dinucleotide scaffold, which more closely resembles the native cap architecture, yielded additional gains in inhibitory activity for specific analogues, consistent with strengthened cap-like binding geometry and an expanded interaction network within the eIF4E binding pocket. Collectively, these results establish a versatile synthetic platform for constructing a diverse cap-analogue library from mononucleotides to dinucleotides and highlight N²/N⁷ substitution patterns that enhance translation inhibition. The compounds described herein may serve as biochemical tools to dissect eIF4E-mediated initiation and provide a basis for developing mechanism-driven modulators of dysregulated translation.

목차 (Table of Contents)

• 1. 초록 영문 (Abstract) 1
• 2. 서론 (Introduction) 3
• 3. 합성 (chemistry) 4
• 3.1. 합성 절차 6
• 3.2. GMP·TEA 및 GDP·TEA의 조제 6
• 3.2.1. N7(2Py)GMP (D1, N7-[(pyridin-2-yl)methyl]guanosine 5′-monophosphate)의 합성 7
• 3.2.2. N7(2Py)GpppG (D1′, N7-[(pyridin-2-yl)methyl]guanosine 5′-triphosphate–5′-guanosine)의 합성 7
• 3.2.3. N7(2TP)GMP (D2, N7-[(thiophen-2-yl)methyl]guanosine 5′-monophosphate)의 합성 8
• 3.2.4. N2His·N7MeGMP (D3, N7-methyl-N2-(histidinylmethyl)guanosine 5′-monophosphate)의 합성 9
• 3.2.5. N2MeIm·N7BnGMP (D4, N7-benzyl-N2-[(1-methyl-1H-imidazol-2-yl)methyl]guanosine 5′-monophosphate)의 합성 10
• 3.2.6. N2MeIm·N7BnGpppG (D4′, N7-benzyl-N2-[(1-methyl-1H-imidazol-2-yl)methyl]guanosine 5′-triphosphate–5′-guanosine)의 합성 11
• 3.2.7. N2MeIm·N7(2TP)GMP (D5, N7-[(thiophen-2-yl)methyl]-N2-[(1-methyl-1H-imidazol-2-yl)methyl]guanosine 5′-monophosphate)의 합성 12
• 3.2.8. N7(2Fu)GMP (D6, N7-[(furan-2-yl)methyl]guanosine 5′-monophosphate)의 합성 13
• 3.2.9. N7(2Fu)GpppG (D6′, N7-[(furan-2-yl)methyl]guanosine 5′-triphosphate–5′-guanosine)의 합성 13
• 3.2.10. N2BocHis·N7BnGMP (D7, N7-benzyl-N2-(Boc-histidinylmethyl)guanosine 5′-monophosphate)의 합성 14
• 3.2.11. N2Uro·N7BnGMP (D8, N7-benzyl-N2-(urocanoylmethyl)guanosine 5′-monophosphate)의 합성 15
• 3.2.12. m7GpppG (reference, N7-Methyl-diguanosine triphosphate)의 합성 16
• 4. 생물학적 평가 (Biology) 17
• 4.1. 단핵산 유도체의 번역 억제력 비교 (Comparative inhibitory potency of mononucleotide analogues) 17
• 4.2. 이핵산 유도체의 번역 억제력 비교 (Comparative inhibitory potency of dinucleotide analogues) 19
• 5. 결론 (Conclusions) 22
• 6. 참고문헌 (References) 24
• 부록 26