Discovery of 6-substituted thieno[2,3-d]pyrimidine analogs as dual inhibitors of glycinamide ribonucleotide formyltransferase and 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase in de novo purine nucleotide biosynthesis in folate receptor expressing human tumors

Wallace-Povirk, Adrianne; Tong, Nian; Wong-Roushar, Jennifer; O’Connor, Carrie; Zhou, Xilin; Hou, Zhanjun; Bao, Xun; García, Gloria Esperanza Castellanos; Li, Jing; Kim, Seongho; Dann, Charles E.; Matherly, Larry H.; Gangjee, Aleem

doi:10.1016/j.bmc.2021.116093

Cited by 10 publications

(36 citation statements)

References 63 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Further, inhibitions of the murine EOCs by all compounds approximated those for IGROV1 human EOC cells. For both murine and human EOC cells, inhibitions were significantly decreased by excess folic acid, establishing their cellular uptake by FRα with secondary uptake by PCFT 10 , 23 .…”

Section: Resultsmentioning

confidence: 90%

“…Our lead analogs (i.e., AGF94 , AGF278 , and AGF283 ; Fig. 1 ) inhibited glycinamide ribonucleotide (GAR) formyltransferase (GARFTase), the first folate-dependent step in the 10 reactions comprising de novo purine biosynthesis, resulting in perturbations in purine precursors and decreased pools of purine nucleotides 10 , 21 – 23 . Anti-tumor activity of this series of compounds including EOC reflects their endocytosis by FRα and/or their facilitative uptake by PCFT 10 , 21 .…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Targeted therapy of pyrrolo[2,3-d]pyrimidine antifolates in a syngeneic mouse model of high grade serous ovarian cancer and the impact on the tumor microenvironment

Wallace-Povirk

Rubinsak

Malysa

et al. 2022

Sci Rep

Self Cite

View full text Add to dashboard Cite

Novel therapies are urgently needed for epithelial ovarian cancer (EOC), the most lethal gynecologic malignancy. In addition, therapies that target unique vulnerabilities in the tumor microenvironment (TME) of EOC have largely been unrealized. One strategy to achieve selective drug delivery for EOC therapy involves use of targeted antifolates via their uptake by folate receptor (FR) proteins, resulting in inhibition of essential one-carbon (C1) metabolic pathways. FRα is highly expressed in EOCs, along with the proton-coupled folate transporter (PCFT); FRβ is expressed on activated macrophages, a major infiltrating immune population in EOC. Thus, there is great potential for targeting both the tumor and the TME with agents delivered via selective transport by FRs and PCFT. In this report, we investigated the therapeutic potential of a novel cytosolic C1 6-substituted pyrrolo[2,3-d]pyrimidine inhibitor AGF94, with selectivity for uptake by FRs and PCFT and inhibition of de novo purine nucleotide biosynthesis, against a syngeneic model of ovarian cancer (BR-Luc) which recapitulates high-grade serous ovarian cancer in patients. In vitro activity of AGF94 was extended in vivo against orthotopic BR-Luc tumors. With late-stage subcutaneous BR-Luc xenografts, AGF94 treatment resulted in substantial anti-tumor efficacy, accompanied by significantly decreased M2-like FRβ-expressing macrophages and increased CD3+ T cells, whereas CD4+ and CD8+ T cells were unaffected. Our studies demonstrate potent anti-tumor efficacy of AGF94 in the therapy of EOC in the context of an intact immune system, and provide a framework for targeting the immunosuppressive TME as an essential component of therapy.

show abstract

Section: Resultsmentioning

confidence: 90%

Section: Introductionmentioning

confidence: 99%

Targeted therapy of pyrrolo[2,3-d]pyrimidine antifolates in a syngeneic mouse model of high grade serous ovarian cancer and the impact on the tumor microenvironment

Wallace-Povirk

Rubinsak

Malysa

et al. 2022

Sci Rep

Self Cite

View full text Add to dashboard Cite

show abstract

“…The data are shown as mean values from at least three experiments. Data for PMX, 1 , 2, 10 , and 11 were published. ,, …”

Section: Resultsmentioning

confidence: 99%

“…Additional chemicals were purchased from commercial sources in the highest available purity. Compounds 1 , 2 , 10 , 11 , AGF23 , and AGF347 were synthesized as previously described. ,,, …”

Section: Biological Studiesmentioning

confidence: 99%

Multitargeted 6-Substituted Thieno[2,3-d]pyrimidines as Folate Receptor-Selective Anticancer Agents that Inhibit Cytosolic and Mitochondrial One-Carbon Metabolism

Tong

Wong-Roushar

Wallace-Povirk

et al. 2023

ACS Pharmacol. Transl. Sci.

Self Cite

View full text Add to dashboard Cite

Multitargeted agents with tumor selectivity result in reduced drug resistance and dose-limiting toxicities. We report 6substituted thieno [2,3-d]pyrimidine compounds (3−9) with pyridine (3, 4), fluorine-substituted pyridine (5), phenyl (6, 7), and thiophene side chains (8, 9), for comparison with unsubstituted phenyl (1, 2) and thiophene side chain (10, 11) containing thieno[2,3-d]pyrimidine compounds. Compounds 3−9 inhibited proliferation of Chinese hamster ovary cells (CHO) expressing folate receptors (FRs) α or β but not the reduced folate carrier (RFC); modest inhibition of CHO cells expressing the proton-coupled folate transporter (PCFT) by 4, 5, 6, and 9 was observed. Replacement of the side-chain 1′,4′-phenyl ring with 2′,5′-pyridyl, or 2′,5′-pyridyl with a fluorine insertion ortho to Lglutamate resulted in increased potency toward FR-expressing CHO cells. Toward KB tumor cells, 4−9 were highly active (IC 50 's from 2.11 to 7.19 nM). By metabolite rescue in KB cells and in vitro enzyme assays, de novo purine biosynthesis was identified as a targeted pathway (at 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase (AICARFTase) and glycinamide ribonucleotide formyltransferase (GARFTase)). Compound 9 was 17-to 882-fold more potent than previously reported compounds 2, 10, and 11 against GARFTase. By targeted metabolomics and metabolite rescue, 1, 2, and 6 also inhibited mitochondrial serine hydroxymethyl transferase 2 (SHMT2); enzyme assays confirmed inhibition of SHMT2. X-ray crystallographic structures were obtained for 4, 5, 9, and 10 with human GARFTase. This series affords an exciting new structural platform for potent multitargeted antitumor agents with FR transport selectivity.

show abstract

“…According to reports, the cellular uptake of 6-substituted pyrrolo [2,3-d] pyrimidines, depend on multiple (folate) transporters like folate receptor alpha (FRα), FRβ, reduced folate carrier (RFC), and proton coupled folate transporter (PCFT). And they found that the role of folylpolyglutamate synthetase (FPGS) in the retention of 6-substituted pyrrolo [2,3-d] pyrimidines intracellularly ( Dekhne et al, 2020 ; Golani et al, 2020 ; Wallace-Povirk et al, 2021 ; Wallace-Povirk et al, 2022 ). Thus, this hypersensitivity maybe explained by the fact that KB cells express high amount of FRα, which may serve as cellular internalization route for glytrexate.…”

Section: Discussionmentioning

confidence: 99%

Pharmacokinetics, bioavailability, and plasma protein binding study of glytrexate, a novel multitarget antifolate

Xiang

Wu²,

Wang³

et al. 2022

Front. Pharmacol.

View full text Add to dashboard Cite

Glytrexate, developed by our team, as a novel multitarget folate antagonist, has inhibitory effects on a variety of cancer cell types, especially KB tumor cells (IC50 0.078 nM), and thus has antitumor drug development prospects. However, its pharmacokinetics and plasma protein binding properties remain unknown. In this study a selective and sensitive liquid chromatography-tandem mass spectrometry (LC‒MS/MS) method was developed and verified to facilitate biological analysis. The bioanalysis method was applied to evaluate the stability, plasma protein binding, and pharmacokinetics of glytrexate. Glytrexate is more stable in human plasma than in rat plasma and in human liver microsomes. The binding of glytrexate to human plasma proteins was higher than that to rat plasma proteins, both of which were less than 30%, suggesting that glytrexate may be at a higher concentration at the pharmacologic target receptor(s) in tissues. Pharmacokinetic characteristics were determined by noncompartmental analysis after administration of single oral (12.5, 25 and 50 mg/kg) and intravenous (2 mg/kg) doses in rats. According to the rat oral pharmacokinetic characteristics, glytrexate had linear dynamics in a dose range of 12.5–50 mg/kg and a poor oral bioavailability of 0.57–1.15%. The investigation revealed that the intravenous half-life, AUC, and Cmax of glytrexate were higher than those of pemetrexed. Pemetrexed is generally produced as an injection preparation. This provides ideas for the development of glytrexate formulations. Therefore, glytrexate injection has clinical application prospects compared to oral administration. This study provides a basis for further investigations into the pharmacological effects and clinical uses of glytrexate.

show abstract

Discovery of 6-substituted thieno[2,3-d]pyrimidine analogs as dual inhibitors of glycinamide ribonucleotide formyltransferase and 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase in de novo purine nucleotide biosynthesis in folate receptor expressing human tumors

Cited by 10 publications

References 63 publications

Targeted therapy of pyrrolo[2,3-d]pyrimidine antifolates in a syngeneic mouse model of high grade serous ovarian cancer and the impact on the tumor microenvironment

Targeted therapy of pyrrolo[2,3-d]pyrimidine antifolates in a syngeneic mouse model of high grade serous ovarian cancer and the impact on the tumor microenvironment

Multitargeted 6-Substituted Thieno[2,3-d]pyrimidines as Folate Receptor-Selective Anticancer Agents that Inhibit Cytosolic and Mitochondrial One-Carbon Metabolism

Pharmacokinetics, bioavailability, and plasma protein binding study of glytrexate, a novel multitarget antifolate

Contact Info

Product

Resources

About