Novel Pyrrolo[3,2- d ]pyrimidine Compounds Target Mitochondrial and Cytosolic One-carbon Metabolism with Broad-spectrum Antitumor Efficacy

• Published in: Molecular cancer therapeutics 2019-10, Vol.18 (10), p.1787-1799
• Main Authors: Dekhne, Aamod S, Shah, Khushbu, Ducker, Gregory S, Katinas, Jade M, Wong-Roushar, Jennifer, Nayeen, Md Junayed, Doshi, Arpit, Ning, Changwen, Bao, Xun, Frühauf, Josephine, Liu, Jenney, Wallace-Povirk, Adrianne, O'Connor, Carrie, Dzinic, Sijana H, White, Kathryn, Kushner, Juiwanna, Kim, Seongho, Hüttemann, Maik, Polin, Lisa, Rabinowitz, Joshua D, Li, Jing, Hou, Zhanjun, Dann, 3rd, Charles E, Gangjee, Aleem, Matherly, Larry H
• Format: Article
• Language: English
• Subjects: Animals; Antineoplastic Agents - chemistry; Antineoplastic Agents - pharmacology; Biosynthetic Pathways - drug effects; Carbon - metabolism; Cell Line, Tumor; CHO Cells; Cricetinae; Cricetulus; Cytosol - drug effects; Cytosol - metabolism; Female; Inhibitory Concentration 50; Metabolomics; Mice, SCID; Mitochondria - drug effects; Mitochondria - metabolism; Purines - biosynthesis; Pyrimidines - chemistry; Pyrimidines - pharmacology; Pyrroles - chemistry; Pyrroles - pharmacology; Xenograft Model Antitumor Assays
• ISSN: 1535-7163; 1538-8514
• DOI: 10.1158/1535-7163.MCT-19-0037

Folate-dependent one-carbon (C1) metabolism is compartmentalized into the mitochondria and cytosol and supports cell growth through nucleotide and amino acid biosynthesis. Mitochondrial C1 metabolism, including serine hydroxymethyltransferase (SHMT) 2, provides glycine, NAD(P)H, ATP, and C1 units for cytosolic biosynthetic reactions, and is implicated in the oncogenic phenotype across a wide range of cancers. Whereas multitargeted inhibitors of cytosolic C1 metabolism, such as pemetrexed, are used clinically, there are currently no anticancer drugs that specifically target mitochondrial C1 metabolism. We used molecular modeling to design novel small-molecule pyrrolo[3,2- ]pyrimidine inhibitors targeting mitochondrial C1 metabolism at SHMT2. antitumor efficacy was established with the lead compounds ( , , ) toward lung, colon, and pancreatic cancer cells. Intracellular targets were identified by metabolic rescue with glycine and nucleosides, and by targeted metabolomics using a stable isotope tracer, with confirmation by assays with purified enzymes. In addition to targeting SHMT2, inhibition of the cytosolic purine biosynthetic enzymes, β-glycinamide ribonucleotide formyltransferase and/or 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase, and SHMT1 was also established. generated significant antitumor efficacy with potential for complete responses against both early-stage and upstage MIA PaCa-2 pancreatic tumor xenografts, providing compelling proof-of-concept for therapeutic targeting of SHMT2 and cytosolic C1 enzymes by this series. Our results establish structure-activity relationships and identify exciting new drug prototypes for further development as multitargeted antitumor agents.