Structure-Activity Relationships of Glutamate Carboxypeptidase II (GCPII) Inhibitors

Ferraris, Dana; Shukla, Krupa H.; Tsukamoto, Takashi

doi:10.2174/092986712799462658

Cited by 37 publications

(38 citation statements)

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“…In the brain and peripheral nervous system, in which it cleaves the abundant dipeptide neurotransmitter N-acetylaspartylglutamate (NAAG) to liberate glutamate (12, 13), the enzyme is referred to as glutamate carboxypeptidase II (GCPII). Multiple classes of potent and selective small-molecule GCPII inhibitors have been synthesized (14–18) and shown to have profound therapeutic effects in a variety of preclinical models of neurological disorders wherein excess glutamate is implicated (18–22). …”

Section: Introductionmentioning

confidence: 99%

FOLH1/GCPII is elevated in IBD patients, and its inhibition ameliorates murine IBD abnormalities

Rais¹,

Jiang²,

Zhai³

et al. 2016

JCI Insight

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Recent gene-profiling analyses showed significant upregulation of the folate hydrolase (FOLH1) gene in the affected intestinal mucosa of patients with inflammatory bowel disease (IBD). The FOLH1 gene encodes a type II transmembrane glycoprotein termed glutamate carboxypeptidase II (GCPII). To establish that the previously reported increased gene expression was functional, we quantified the glutamate carboxypeptidase enzymatic activity in 31 surgical specimens and report a robust 2.8- to 41-fold increase in enzymatic activity in the affected intestinal mucosa of IBD patients compared with an uninvolved area in the same patients or intestinal mucosa from healthy controls. Using a human-to-mouse approach, we next showed a similar enzymatic increase in two well-validated IBD murine models and evaluated the therapeutic effect of the potent FOLH1/ GCPII inhibitor 2-phosphonomethyl pentanedioic acid (2-PMPA) (IC50 = 300 pM). In the dextran sodium sulfate (DSS) colitis model, 2-PMPA inhibited the GCPII activity in the colonic mucosa by over 90% and substantially reduced the disease activity. The significance of the target was confirmed in FOLH1−/− mice who exhibited resistance to DSS treatment. In the murine IL-10−/− model of spontaneous colitis, daily 2-PMPA treatment also significantly reduced both macroscopic and microscopic disease severity. These results provide the first evidence of FOLH1/GCPII enzymatic inhibition as a therapeutic option for IBD.

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Section: Introductionmentioning

confidence: 99%

FOLH1/GCPII is elevated in IBD patients, and its inhibition ameliorates murine IBD abnormalities

Rais¹,

Jiang²,

Zhai³

et al. 2016

JCI Insight

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“…Presently, the two major categories of GCPII-specific inhibitors that exist are either analogs of NAAG (the GCPII substrate) or derivatives of glutamic acid (the reaction product). 12,13 Consequently, both the chemical space tapped by such compounds and the diversity of GCPII-specific compounds are limited. For example, NAAG- or glutamate-based inhibitors are highly polar substances, with mitigated penetration into the neuronal compartment.…”

Section: Introductionmentioning

confidence: 99%

Development of a High-Throughput Fluorescence Polarization Assay to Identify Novel Ligands of Glutamate Carboxypeptidase II

et al. 2012

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Glutamate carboxypeptidase II (GCPII) is an important target for therapeutic and diagnostic interventions aimed at prostate cancer and neurologic disorders. Here we describe the development and optimization of a high-throughput screening (HTS) assay based on fluorescence polarization (FP) that facilitates the identification of novel scaffolds inhibiting GCPII. First, we designed and synthesized a fluorescence probe based on a urea-based inhibitory scaffold covalently linked to a Bodipy TMR fluorophore (TMRGlu). Next, we established and optimized conditions suitable for HTS and evaluated the assay robustness by testing the influence of a variety of physicochemical parameters (e.g., pH, temperature, time) and additives. Using known GCPII inhibitors, the FP assay was shown to be comparable to benchmark assays established in the field. Finally, we evaluated the FP assay by HTS of a 20 000–compound library. The novel assay presented here is robust, highly reproducible (Z′ = 0.82), inexpensive, and suitable for automation, thus providing an excellent platform for HTS of small-molecule libraries targeting GCPII.

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“…Clinical PSMA imaging reports generally describe the use of radiolabeled high-affinity targeting moieties based on glutamateurea, such as the inhibitor motif glutamic acid-urea-lysine (EuK) (8,9), which are applied in a microdose regime (#100 mg/patient). The molecular binding interactions of glutamate-urea moieties to the PSMA protein have been studied extensively (10). The glutamateureido derivatives coordinate specifically with the binuclear zinc present deep within the PSMA protein.…”

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confidence: 99%

Hybrid Tracers Based on Cyanine Backbones Targeting Prostate-Specific Membrane Antigen: Tuning Pharmacokinetic Properties and Exploring Dye–Protein Interaction

et al. 2019

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Prostate cancer surgery is currently being revolutionized by the use of prostate-specific membrane antigen (PSMA)-targeted radiotracers, for example, 99m Tc-labeled PSMA tracer analogs for radioguided surgery. The purpose of this study was to develop a second-generation 99m Tc-labeled PSMA-targeted tracer incorporating a fluorescent dye. Methods: Several PSMA-targeted hybrid tracers were synthesized: glutamic acid-urea-lysine (EuK)-Cy5-mas 3 , EuK-(SO 3)Cy5-mas 3 , EuK-Cy5(SO 3)-mas 3 , EuK-(Ar)Cy5-mas 3 , and EuK-Cy5(Ar)-mas 3 ; the Cy5 dye acts as a functional backbone between the EuK targeting vector and the 2-mercaptoacetyl-seryl-seryl-seryl (mas 3) chelate to study the dye's interaction with PSMA's amphipathic entrance funnel. The compounds were evaluated for their photophysical and chemical properties and PSMA affinity. After radiolabeling with 99m Tc, we performed in vivo SPECT imaging, biodistribution, and fluorescence imaging on BALB/c nude mice with orthotopically transplanted PC346C tumors. Results: The dye composition influenced the photophysical properties (brightness range 0.3-1.5 • 10 4 M −1 • cm −1), plasma protein interactions (range 85.0% ± 2.3%-90.7% ± 1.3% bound to serum, range 76% ± 0%-89% ± 6% stability in serum), PSMA affinity (half-maximal inhibitory concentration [IC 50 ] range 19.2 ± 5.8-175.3 ± 61.1 nM) and in vivo characteristics (tumorto-prostate and tumor-to-muscle ratios range 0.02 ± 0.00-154.73 ± 28.48 and 0.46 ± 0.28-5,157.50 ± 949.17, respectively; renal, splenic, and salivary retention). Even though all tracer analogs allowed tumor identification with SPECT and fluorescence imaging, 99m Tc-EuK-(SO 3)Cy5-mas 3 had the most promising properties (e.g., half-maximal inhibitory concentration, 19.2 ± 5.8, tumor-to-muscle ratio, 5,157.50 ± 949.17). Conclusion: Our findings demonstrate the intrinsic integration of a fluorophore in the pharmacophore in PSMA-targeted smallmolecule tracers. In this design, having 1 sulfonate on the indole moiety adjacent to EuK (99m Tc-EuK-(SO 3)Cy5-mas 3) yielded the most promising tracer candidate for imaging of PSMA.

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Structure-Activity Relationships of Glutamate Carboxypeptidase II (GCPII) Inhibitors

Cited by 37 publications

References 0 publications

FOLH1/GCPII is elevated in IBD patients, and its inhibition ameliorates murine IBD abnormalities

FOLH1/GCPII is elevated in IBD patients, and its inhibition ameliorates murine IBD abnormalities

Development of a High-Throughput Fluorescence Polarization Assay to Identify Novel Ligands of Glutamate Carboxypeptidase II

Hybrid Tracers Based on Cyanine Backbones Targeting Prostate-Specific Membrane Antigen: Tuning Pharmacokinetic Properties and Exploring Dye–Protein Interaction

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