Structure–Activity Studies of 3,9-Diazaspiro[5.5]undecane-Based γ-Aminobutyric Acid Type A Receptor Antagonists with Immunomodulatory Effect

Bavo, Francesco; de-Jong, Heleen; Petersen, Jonas; Falk-Petersen, Christina Birkedahl; Löffler, Rebekka; Sparrow, Emma L; Rostrup, Frederik; Eliasen, Jannik Nicklas; Wilhelmsen, Kristine S.; Barslund, Kasper; Bundgaard, Christoffer; Nielsen, Birgitte; Kristiansen, Uffe; Wellendorph, Petrine; Bogdanov, Yury; Frølund, Bente

doi:10.1021/acs.jmedchem.1c00290

Cited by 4 publications

(3 citation statements)

References 59 publications

(144 reference statements)

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“…[ 3 H]muscimol binding assay in rat cortical synaptic membranes as previously described [35]. *Data taken from Lie et al [17].…”

Section: Tablementioning

confidence: 99%

Structural and stereochemical determinants for hGAT3 inhibition: development of novel conformationally constrained and substituted analogs of (S)-isoserine.

Bavo,

Kickinger,

Lie

et al. 2023

Preprint

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The GABA transporter 3 (GAT3) is a member of the GABA transporter (GAT) family proposed to have a role in regulating tonic inhibition. The GAT3-preferring substrate (S)-isoserine has shown beneficial effects in a mouse model of stroke accompanied by an increased GAT3 expression, indicating a molecular mechanism mediated by GAT3. However, (S)-isoserine is not ideally suited for in vivo studies due to a lack of selectivity and brain permeability. To elucidate the structural determinants of (S)-isoserine for GAT3 inhibition, and to optimize and inform further ligand development, we here present the design, synthesis and pharmacological evaluation of a series of conformationally constrained isoserine analogues with defined stereochemistry. Using [3H]GABA uptake assays at recombinant human GAT3, we identified the azetidine and pyrrolidine analogs ((S,S)-6a and (S,S)-7a) as the most potent inhibitors. To further elaborate on the selectivity profile both compounds were tested at all GATs, the taurine transporter (TauT) and GABAA receptors. Although (S,S)-6a and (S,S)-7a are comparable to (S)-isoserine with respect to potency, the selectivity versus the taurine transporter was significantly improved (at least 6 and 53 times more activity at hGAT3, respectively). A subsequent comprehensive structure-activity study showed that different connectivity approaches, stereochemical variations, simple or larger α- and N- substituents, and even minor size enlargement of the alicyclic ring all abrogated GAT3 inhibition, indicating very strict stereochemical and size requirements. The observed structure activity relationships may guide future ligand optimization and the novel ligands ((S,S)-6a and (S,S)-7a) can serve as valuable tools to validate the proposed GAT3-mediated effect of (S)-isoserine such as in functional recovery after stroke and thus help corroborate the relevance of targeting GAT3 and tonic inhibition in relevant brain pathologies.

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“…[ 3 H]muscimol binding assay in rat cortical synaptic membranes as previously described [35]. *Data taken from Lie et al [17].…”

Section: Tablementioning

confidence: 99%

Structural and stereochemical determinants for hGAT3 inhibition: development of novel conformationally constrained and substituted analogs of (S)-isoserine.

Bavo,

Kickinger,

Lie

et al. 2023

Preprint

Self Cite

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“…In particular, the immunoassays based on enzyme-linked immunosorbent assay (ELISA) [ 2 , 7 ] and chemiluminescence immunoassay (CLIA) [ 8 ] are currently the most commonly used protein assays, in which ELISA serves as the gold standard for protein detection in the fields of clinical diagnosis and biosafety [ 9 ]. However, the levels of protein biomarkers in clinical samples are generally very low, while a large amount of matrix interference exists in samples [ 10 ], so ELISA assays in clinical use are generally limited by their sensitivity, reliability, and specificity [ 11 ]. Therefore, it is urgent to develop new methods for the rapid, sensitive, portable, and highly specific detection of protein biomarkers.…”

Section: Introductionmentioning

confidence: 99%

Recent Advances in CRISPR/Cas-Based Biosensors for Protein Detection

et al. 2022

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CRISPR is an acquired immune system found in prokaryotes that can accurately recognize and cleave foreign nucleic acids, and has been widely explored for gene editing and biosensing. In the past, CRISPR/Cas-based biosensors were mainly applied to detect nucleic acids in the field of biosensing, and their applications for the detection of other types of analytes were usually overlooked such as small molecules and disease-related proteins. The recent work shows that CRISPR/Cas biosensors not only provide a new tool for protein analysis, but also improve the sensitivity and specificity of protein detections. However, it lacks the latest review to summarize CRISPR/Cas-based biosensors for protein detection and elucidate their mechanisms of action, hindering the development of superior biosensors for proteins. In this review, we summarized CRISPR/Cas-based biosensors for protein detection based on their mechanism of action in three aspects: antibody-assisted CRISPR/Cas-based protein detection, aptamer-assisted CRISPR/Cas-based protein detection, and miscellaneous CRISPR/Cas-based methods for protein detection, respectively. Moreover, the prospects and challenges for CRISPR/Cas-based biosensors for protein detection are also discussed.

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“…Propofol is a type A γ -aminobutyric acid receptor (GABA A R) agonist that exists in the peripheral tissue and has become a tumor-promoting molecule that controls the growth of tumor cells [ 13 , 14 ]. GABA A Rs could enhance growth and promote migration and invasion of papillary thyroid cancer [ 15 ].…”

Section: Introductionmentioning

confidence: 99%

Propofol enhances stem-like properties of glioma via GABAAR‐dependent Src modulation of ZDHHC5-EZH2 palmitoylation mechanism

Fan

Gong

et al. 2022

Stem Cell Res Ther

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Background Propofol is a commonly used anesthetic. However, its effects on glioma growth and recurrence remain largely unknown. Methods The effect of propofol on glioma growth was demonstrated by a series of in vitro and in vivo experiments (spheroidal formation assay, western blotting, and xenograft model). The acyl-biotin exchange method and liquid chromatography-mass spectrometry assays identified palmitoylation proteins mediated by the domain containing the Asp-His-His-Cys family. Western blotting, co-immunoprecipitation, quantitative real-time polymerase chain reaction, co-immunoprecipitation, chromatin immunoprecipitation, and luciferase reporter assays were used to explore the mechanisms of the γ-aminobutyric acid receptor (GABAAR)/Src/ZDHHC5/EZH2 signaling axis in the effects of propofol on glioma stem cells (GSCs). Results We found that treatment with a standard dose of propofol promoted glioma growth in nude mice compared with control or low-dose propofol. Propofol-treated GSCs also led to larger tumor growth in nude mice than did vector-treated tumors. Mechanistically, propofol enhances the stem-like properties of gliomas through GABAAR to increase Src expression, thereby enhancing the palmitoylation of ZDHHC5-mediated EZH2 and Oct4 expression. Conclusion These results demonstrate that propofol may promote glioma growth through the GABAAR-Src-ZDHHC5-EZH2 mechanism and are helpful in guiding the clinical use of propofol to obtain a better patient prognosis after the surgical resection of tumors.

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Structure–Activity Studies of 3,9-Diazaspiro[5.5]undecane-Based γ-Aminobutyric Acid Type A Receptor Antagonists with Immunomodulatory Effect

Cited by 4 publications

References 59 publications

Structural and stereochemical determinants for hGAT3 inhibition: development of novel conformationally constrained and substituted analogs of (S)-isoserine.

Structural and stereochemical determinants for hGAT3 inhibition: development of novel conformationally constrained and substituted analogs of (S)-isoserine.

Recent Advances in CRISPR/Cas-Based Biosensors for Protein Detection

Propofol enhances stem-like properties of glioma via GABAAR‐dependent Src modulation of ZDHHC5-EZH2 palmitoylation mechanism

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