title: Tumor Targeting with Novel 6-Substituted Pyrrolo [2,3-d] Pyrimidine Antifolates with Heteroatom Bridge Substitutions via Cellular Uptake by Folate Receptor α and the Proton-Coupled Folate Transporter and Inhibition of de Novo Purine Nucleotide Biosynthesis.
abstract: Targeted antifolates with heteroatom replacements of the carbon vicinal to the phenyl ring in 1 by N (4), O (8), or S (9), or with N-substituted formyl (5), acetyl (6), or trifluoroacetyl (7) moieties, were synthesized and tested for selective cellular uptake by folate receptor (FR) α and β or the proton-coupled folate transporter. Results show increased in vitro antiproliferative activity toward engineered Chinese hamster ovary cells expressing FRs by 4-9 over the CH2 analogue 1. Compounds 4-9 inhibited de novo purine biosynthesis and glycinamide ribonucleotide formyltransferase (GARFTase). X-ray crystal structures for 4 with FRα and GARFTase showed that the bound conformations of 4 required flexibility for attachment to both FRα and GARFTase. In mice bearing IGROV1 ovarian tumor xenografts, 4 was highly efficacious. Our results establish that heteroatom substitutions in the 3-atom bridge region of 6-substituted pyrrolo[2,3-d]pyrimidines related to 1 provide targeted antifolates that warrant further evaluation as anticancer agents.
authors.affiliation: Division of Medicinal Chemistry, Graduate School of Pharmaceutical Sciences, Duquesne University , 600 Forbes Avenue, Pittsburgh, Pennsylvania 15282, United States.
authors.forename: Lalit K
authors.initials: LK
authors.lastname: Golani
authors.affiliation: Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute , 110 East Warren Avenue, Detroit, Michigan 48201, United States.
authors.forename: Adrianne
authors.initials: A
authors.lastname: Wallace-Povirk
authors.affiliation: Department of Chemistry, Indiana University , Bloomington, Indiana 47405, United States.
authors.forename: Siobhan M
authors.initials: SM
authors.lastname: Deis
authors.affiliation: Department of Chemistry, Indiana University , Bloomington, Indiana 47405, United States.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Wong
authors.affiliation: Laboratory of Structural Sciences and Laboratory of Structural Biology and Biochemistry, Van Andel Research Institute , 333 Bostwick Avenue NE, Grand Rapids, Michigan 49503, United States.
authors.forename: Jiyuan
authors.initials: J
authors.lastname: Ke
authors.affiliation: Laboratory of Structural Sciences and Laboratory of Structural Biology and Biochemistry, Van Andel Research Institute , 333 Bostwick Avenue NE, Grand Rapids, Michigan 49503, United States.
authors.forename: Xin
authors.initials: X
authors.lastname: Gu
authors.affiliation: Division of Medicinal Chemistry, Graduate School of Pharmaceutical Sciences, Duquesne University , 600 Forbes Avenue, Pittsburgh, Pennsylvania 15282, United States.
authors.forename: Sudhir
authors.initials: S
authors.lastname: Raghavan
authors.affiliation: Department of Oncology, Wayne State University School of Medicine , Detroit, Michigan 48201, United States.
authors.forename: Mike R
authors.initials: MR
authors.lastname: Wilson
authors.affiliation: Division of Medicinal Chemistry, Graduate School of Pharmaceutical Sciences, Duquesne University , 600 Forbes Avenue, Pittsburgh, Pennsylvania 15282, United States.
authors.forename: Xinxin
authors.initials: X
authors.lastname: Li
authors.affiliation: Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute , 110 East Warren Avenue, Detroit, Michigan 48201, United States.|Department of Oncology, Wayne State University School of Medicine , Detroit, Michigan 48201, United States.
authors.forename: Lisa
authors.initials: L
authors.lastname: Polin
authors.affiliation: Laboratory of Structural Sciences and Laboratory of Structural Biology and Biochemistry, Van Andel Research Institute , 333 Bostwick Avenue NE, Grand Rapids, Michigan 49503, United States.
authors.forename: Parker W
authors.initials: PW
authors.lastname: de Waal
authors.affiliation: Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute , 110 East Warren Avenue, Detroit, Michigan 48201, United States.|Department of Oncology, Wayne State University School of Medicine , Detroit, Michigan 48201, United States.
authors.forename: Kathryn
authors.initials: K
authors.lastname: White
authors.affiliation: Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute , 110 East Warren Avenue, Detroit, Michigan 48201, United States.|Department of Oncology, Wayne State University School of Medicine , Detroit, Michigan 48201, United States.
authors.forename: Juiwanna
authors.initials: J
authors.lastname: Kushner
authors.affiliation: Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute , 110 East Warren Avenue, Detroit, Michigan 48201, United States.
authors.forename: Carrie
authors.initials: C
authors.lastname: O'Connor
authors.affiliation: Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute , 110 East Warren Avenue, Detroit, Michigan 48201, United States.|Department of Oncology, Wayne State University School of Medicine , Detroit, Michigan 48201, United States.
authors.forename: Zhanjun
authors.initials: Z
authors.lastname: Hou
authors.affiliation: Laboratory of Structural Sciences and Laboratory of Structural Biology and Biochemistry, Van Andel Research Institute , 333 Bostwick Avenue NE, Grand Rapids, Michigan 49503, United States.|Key Laboratory of Receptor Research, VARI-SIMM Center, Center for Structure and Function of Drug Targets, Shanghai Institute of Materia Medica, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences , Shanghai 201203, People's Republic of China.
authors.forename: H Eric
authors.initials: HE
authors.lastname: Xu
authors.affiliation: Laboratory of Structural Sciences and Laboratory of Structural Biology and Biochemistry, Van Andel Research Institute , 333 Bostwick Avenue NE, Grand Rapids, Michigan 49503, United States.
authors.forename: Karsten
authors.initials: K
authors.lastname: Melcher
authors.affiliation: Department of Chemistry, Indiana University , Bloomington, Indiana 47405, United States.
authors.forename: Charles E
authors.initials: CE
authors.lastname: Dann
authors.affiliation: Molecular Therapeutics Program, Barbara Ann Karmanos Cancer Institute , 110 East Warren Avenue, Detroit, Michigan 48201, United States.|Department of Oncology, Wayne State University School of Medicine , Detroit, Michigan 48201, United States.|Department of Pharmacology, Wayne State University School of Medicine , Detroit, Michigan 48201, United States.
authors.forename: Larry H
authors.initials: LH
authors.lastname: Matherly
authors.affiliation: Division of Medicinal Chemistry, Graduate School of Pharmaceutical Sciences, Duquesne University , 600 Forbes Avenue, Pittsburgh, Pennsylvania 15282, United States.
authors.forename: Aleem
authors.initials: A
authors.lastname: Gangjee
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D045744:Cell Line, Tumor; D018360:Crystallography, X-Ray; D004354:Drug Screening Assays, Antitumor; D005260:Female; D058975:Folate Receptor 1 / Q000378:metabolism*; D005493:Folic Acid Antagonists / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D064593:Heterografts; D006801:Humans; D016513:Mice, SCID; D062105:Molecular Docking Simulation; D009368:Neoplasm Transplantation; D051026:Phosphoribosylglycinamide Formyltransferase / Q000037:antagonists & inhibitors; D058979:Proton-Coupled Folate Transporter / Q000378:metabolism*; D011685:Purine Nucleotides / Q000037:antagonists & inhibitors* / Q000096:biosynthesis; D011743:Pyrimidines / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D011758:Pyrroles / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology; D013329:Structure-Activity Relationship
chemical_list: D000970:Antineoplastic Agents; D058975:Folate Receptor 1; D005493:Folic Acid Antagonists; D058979:Proton-Coupled Folate Transporter; D011685:Purine Nucleotides; D011743:Pyrimidines; D011758:Pyrroles; D051026:Phosphoribosylglycinamide Formyltransferase