Discovery of Novel Phthalazinone Derivatives as Potent Ectonucleotide Pyrophosphatase/Phosphodie sterase-1 (ENPP1) Inhibitor for Cancer Immunotherapy
- 주제(키워드) ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) , 2’ , 3’-cyclic guanosine monophosphate–adenosine monophosphate (cGAMP) , cyclic guanosine monophosphate–adenosine monophosphate synthase–stimulator of interferon genes (cGAS–STING) , innate immune system , phthalazinone inhibitor
- 주제(DDC) 547
- 발행기관 아주대학교
- 지도교수 최준원
- 발행년도 2023
- 학위수여년월 2023. 2
- 학위명 석사
- 학과 및 전공 일반대학원 분자과학기술학과
- 실제URI http://www.dcollection.net/handler/ajou/000000032656
- 본문언어 한국어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
The cyclic guanosine monophosphate–adenosine monophosphate synthase–stimulator of interferon genes (cGAS–STING) pathway is an important component of the innate immune system. In this signaling pathway, 2’,3’-cyclic guanosine monophosphate–adenosine monophosphate (cGAMP) acts as a STING agonist. However, ectonucleotide pyrophosphatase/phosphodiesterase 1 (ENPP1) easily hydrolyzes cGAMP. The inhibition of STING activation via cGAMP hydrolysis induces immune suppression in the tumor microenvironment, thereby decreasing the effectiveness of cancer immunotherapy. We designed, synthesized, and characterized a small-molecule ENPP1 inhibitor that potently stimulates immune activation. All synthesized compounds had a sulfamide head group for binding to zinc in the catalytic domain of ENPP1 and a phthalazinone scaffold that served as the pharmacophore. An AMP-Glo assay was carried out using cGAMP to assess the activity of the ENPP1 inhibitors. To determine the potency of the synthesized compounds, a luciferase reporter assay was performed to determine the expression levels of the interferon genes in HCT116 and THP-1 cells. Compound 7f displayed relatively excellent activity against ENPP1 at the molecular and cellular levels, with IC50 values <100 nM. Further, the oral administration of 7f significantly decreased the tumor volume in a CT26 murine colorectal carcinoma model. Our in vitro and in vivo assays suggest that 7f can be applied in cancer immunotherapy via ENPP1 inhibition.
more목차
1. Introduction 1
1.1. Cancer immunotherapy and the cGAS–STING pathway 1
1.2. ENPP1 as a therapeutic target in cancer 3
1.3. ENPP1 inhibitors for cancer immunotherapy 4
2. Results and Discussion 7
2.1. Synthesis of the phthalazinone derivatives 7
2.2. Structure-activity relationship 13
2.3. Biological evaluation 17
2.4. Docking studies 21
2.5. CYP inhibition, metabolic stability, and PK profiling 21
2.6. In vivo efficacy 23
3. Conclusions 24
4. Experimental Section 25
4.1. General information 25
4.2. Synthetic procedure 26
5. References 134
6. 1H and 13C NMR spectra 144
