compound gdaz 3  (MedChemExpress)

Journal: Advanced Science

Article Title: HSP70 Interactome‐Mediated Proteolysis Targeting Chimera (HSP70‐PROTAC) for Ferroptosis‐Driven Cancer Treatment

doi: 10.1002/advs.202513655

Figure Lengend Snippet: The reported active GPX4‐targeting degraders and schematic diagram of HSP70‐PROTAC to degrade POI via UPS and CMA. A) Structure of representative CRBN‐based GPX4‐targeting PROTAC dGPX4. B) Structure of representative VHL‐based GPX4‐targeting PROTAC 8e. C) Structure of representative HSP90‐based GPX4‐targeting HIM‐PROTAC GDCNF‐11. D) Schematic diagram of heat shock protein 70 interactome‐mediated proteolysis targeting chimera (HSP70‐PROTAC) to degrade POI via USP and CMA dual processes. E) The structures of the synthesized GPX4‐targeting HSP70‐PROTACs including GDAz‐3.

Article Snippet: The proteomics data were generated by Shanghai Bioprofile Co., Ltd. After treatment with compound GDAz‐3 (2 μM) or DMSO for 18 h, the samples were collected.

Techniques: Synthesized

Journal: Advanced Science

Article Title: HSP70 Interactome‐Mediated Proteolysis Targeting Chimera (HSP70‐PROTAC) for Ferroptosis‐Driven Cancer Treatment

doi: 10.1002/advs.202513655

Figure Lengend Snippet: Characterization of GDAz‐3 as a potent GPX4 degrader. A) The preliminary screening of GPX4 degradation effect of the synthesized HSP70‐PROTACs after treatment with different concentrations (0.1, 0.3, 1, and 3 µM) for 24 h in HT1080 cells (calculated using Image J software and the respective DMSO treatment group:100%). B) The representative protein band and the DC 50 curve of compound GDAz‐3 in degrading GPX4 at various concentrations in HT1080 cells after treatment for 24 h as determined by western blot (n = 3). C) The fluorescence imaging and the DC 50 curve of compound GDAz‐3 in degrading GPX4 at various concentrations in HT1080 cells after treatment for 24 h as determined by In‐Cell Western. (D) The mRNA levels of GPX4 were detected by quantitative real‐time PCR assay after treatment with GDAz‐3 for 24 h at the designed concentrations (n = 3). E) The protein bands of compounds CHX or in combination with GDAz‐3 in degrading GPX4 for the designated time. F) The GPX4 protein level was determined at the indicated time‐points after treatment with GDAz‐3 (n = 3). G) Sustained cellular degradative activity induced by GDAz‐3 (1 µM) upon washout in HT1080 cells. Data represent mean ± SD and analyzed by one‐way ANOVA with Dunnett's multiple comparison test (C, compared to vehicle group; ns: not significant).

Article Snippet: The proteomics data were generated by Shanghai Bioprofile Co., Ltd. After treatment with compound GDAz‐3 (2 μM) or DMSO for 18 h, the samples were collected.

Techniques: Synthesized, Software, Western Blot, Fluorescence, Imaging, In-Cell ELISA, Real-time Polymerase Chain Reaction, Activity Assay, Comparison

Journal: Advanced Science

Article Title: HSP70 Interactome‐Mediated Proteolysis Targeting Chimera (HSP70‐PROTAC) for Ferroptosis‐Driven Cancer Treatment

doi: 10.1002/advs.202513655

Figure Lengend Snippet: Both UPS and CMA processes were involved in HSP70‐PROTAC‐mediated GPX4 degradation. A,B) Western blot analysis of GPX4 in HT1080 cells after treatment with compounds as indicated for 24 h. C) The ubiquitination assay of GPX4 after treatment with GDAz‐3 (1 µM) for 12 h. D) Western blot analysis of GPX4 in CHIP‐knockdown HT1080 cells after treatment with GDAz‐3 for 24 h. E) Pre‐treatment with MLN4924 (2.5 µM) for 2 h before the addition of GDAz‐3 for 12 h to prevent Cullin‐dependent ubiquitination and GPX4 degradation. F) Transmission electron microscopy images of autophagosomes in HT1080 cells treated with DMSO and GDAz‐3 (1 µM) for 24 h (scale bar = 1 µm). G,H) Western blot analysis of the level of GPX4 in HT1080 cells after treatment with compounds as indicated for 24 h. I) Western blot analysis of GPX4 in LAMP2A‐knockdown HT1080 cells after treatment with GDAz‐3 for 24 h. J) Representative confocal fluorescence images of HT1080 cells treated with GDAz‐3 (1 µM) for 24 h (scale bar = 2.5 µm).

Article Snippet: The proteomics data were generated by Shanghai Bioprofile Co., Ltd. After treatment with compound GDAz‐3 (2 μM) or DMSO for 18 h, the samples were collected.

Techniques: Western Blot, Ubiquitin Proteomics, Knockdown, Transmission Assay, Electron Microscopy, Fluorescence

Journal: Advanced Science

Article Title: HSP70 Interactome‐Mediated Proteolysis Targeting Chimera (HSP70‐PROTAC) for Ferroptosis‐Driven Cancer Treatment

doi: 10.1002/advs.202513655

Figure Lengend Snippet: Hsc70‐CHIP‐BAG3 chaperone complex participates in GDAz‐3‐mediated GPX4 degradation. A) Chemical modification of GDAz‐3 and western blot analysis of GPX4 in HT1080 cells after treatment with three negative compounds GDAz‐Neg1, GDAz‐Neg2, and GDAz‐OH (unable to bind with HSP70 or GPX4) for 24 h. B) Western blot analysis of GPX4 in HT1080 cells after treatment with compounds as indicated for 24 h. C) Western blot analysis of GPX4 in Hsc70‐knockdown HT1080 cells after treatment with GDAz‐3 for 24 h. D) Western blot analysis of GPX4 in BAG3‐knockdown HT1080 cells after treatment with GDAz‐3 for 24 h. E,F) Western blot analysis of GPX4 in HT1080 cells after treatment with compounds as indicated for 24 h.

Article Snippet: The proteomics data were generated by Shanghai Bioprofile Co., Ltd. After treatment with compound GDAz‐3 (2 μM) or DMSO for 18 h, the samples were collected.

Techniques: Modification, Western Blot, Knockdown

Journal: Advanced Science

Article Title: HSP70 Interactome‐Mediated Proteolysis Targeting Chimera (HSP70‐PROTAC) for Ferroptosis‐Driven Cancer Treatment

doi: 10.1002/advs.202513655

Figure Lengend Snippet: Verification of the ternary complex of Hsc70/GDAz‐3/GPX4. A) Chemical structure of GDAz‐biotin. B) Western blot analysis of GPX4 in HT1080 cells after treatment with GDAz‐biotin for 24 h. C) The interactions between Hsc70 and GPX4 mediated by GDAz‐3 were determined by pull‐down assay. D) SPR analysis of the interaction between immobilized Hsc70 and GDAz‐3. E) Binding affinity of GDAz‐3 to the immobilized Hsc70 protein in the presence of GPX4 protein. F) Computational docking simulation of the ternary complex of Hsc70/GDAz‐3/GPX4 (yellow dashed lines: hydrogen bonds). G) RMSD curve of the ternary structure of Hsc70/GDAz‐3/GPX4 in 500 ns MD simulations.

Article Snippet: The proteomics data were generated by Shanghai Bioprofile Co., Ltd. After treatment with compound GDAz‐3 (2 μM) or DMSO for 18 h, the samples were collected.

Techniques: Western Blot, Pull Down Assay, Binding Assay

Journal: Advanced Science

Article Title: HSP70 Interactome‐Mediated Proteolysis Targeting Chimera (HSP70‐PROTAC) for Ferroptosis‐Driven Cancer Treatment

doi: 10.1002/advs.202513655

Figure Lengend Snippet: Degradation of GPX4 by GDAz‐3 triggers ferroptosis. A) Cytotoxicity of all the synthesized HSP70‐PROTACs including positive control drugs against HT1080 cells (data are presented as mean ± SD). B) Ferroptosis selectivity of GDAz‐3. HT1080 cells were incubated with the tested compounds or in combination with 2 µM Fer‐1 for 48 h. C) Western blot analysis of GPX1 and SLC7A11 in HT1080 cells after treatment with GDAz‐3 for 24 h. D) Confocal laser scanning microscopy imaging of intracellular total ROS upon treatment with the specified agents at 1 µM for 24 h. E) Confocal laser scanning microscopy imaging of intracellular LPO upon treatments with the specified agents at the designated concentration for 24 h. F) Flow cytometry analysis for intracellular LPO of HT1080 cells treated with the specified agents at the designed concentrations at the designated concentration for 12 h (n =3). G) Determination of the content of intracellular MDA, GSH, and Fe 2+ after treatment with various concentrations of GDAz‐3 for 24 h (n =3). H) Transmission electron microscopy images of HT1080 cells treated with DMSO, and GDAz‐3 (1 µM) for 24 h; red arrow: decreased or absent mitochondrial crests, or broken outer membrane. Data represent mean ± SD. * p < 0.05, ** p <0.01, *** p < 0.001, **** p < 0.0001 compared to control group by one‐way ANOVA with Dunnett's multiple comparisons; ns: not significant.

Article Snippet: The proteomics data were generated by Shanghai Bioprofile Co., Ltd. After treatment with compound GDAz‐3 (2 μM) or DMSO for 18 h, the samples were collected.

Techniques: Synthesized, Positive Control, Incubation, Western Blot, Confocal Laser Scanning Microscopy, Imaging, Concentration Assay, Flow Cytometry, Transmission Assay, Electron Microscopy, Membrane, Control

Journal: Advanced Science

Article Title: HSP70 Interactome‐Mediated Proteolysis Targeting Chimera (HSP70‐PROTAC) for Ferroptosis‐Driven Cancer Treatment

doi: 10.1002/advs.202513655

Figure Lengend Snippet: PK and antitumor evaluation of GDAz‐3 in vivo. A) Concentration‐time curve and PK parameters of GDAz‐3 (n = 3). B) Schematics of in vivo antitumor efficacy study of GDAz‐3 (n = 6 per group). C) Tumor growth curves of HT1080 tumor‐bearing mice subjected to diverse treatments (the data were not presented once the mouse died). D) Changes in the body weights of mice over the treatment period (the data were not presented once the mouse died). (E) Image of the subcutaneous tumor of xenografts in different groups of nude mice; F) Comparison of tumor weight in mice from different groups; G) representative immunohistochemistry staining analysis of GPX4 and 4‐HNE from different groups. H) Pharmacodynamic analysis of GPX4 protein in the HT1080 xenograft tumors in nude mice. Mice‐bearing xenograft tumors were treated with a single administration of either vehicle control or GDAz‐3 at 50 mg kg −1 via intraperitoneal injection, and the mice were sacrificed at the indicated time points. Tumor tissues were harvested for western blotting analysis. I) Quantitative analysis of GDAz‐3 concentration in tumor, and other major organ tissues of the HT1080 xenograft tumors in nude mice after intraperitoneal injection at 12 h (n = 1). Data represent mean ± SEM. * p < 0.05, ** p <0.01, *** p < 0.001, **** p < 0.0001 compared to vehicle group by one‐way ANOVA with Dunnett's multiple comparisons (B and E).

Article Snippet: The proteomics data were generated by Shanghai Bioprofile Co., Ltd. After treatment with compound GDAz‐3 (2 μM) or DMSO for 18 h, the samples were collected.

Techniques: In Vivo, Concentration Assay, Comparison, Immunohistochemistry, Staining, Control, Injection, Western Blot

Journal: Advanced Science

Article Title: HSP70 Interactome‐Mediated Proteolysis Targeting Chimera (HSP70‐PROTAC) for Ferroptosis‐Driven Cancer Treatment

doi: 10.1002/advs.202513655

Figure Lengend Snippet: Potential Applicability of HSP70‐PROTAC. A) Western blot analysis of GPX4 in HT1080 cells after treatment with reported GPX4 degraders (1 µM) for 24 h. B,C) Western blot analysis of GPX4 in CRBN‐ or VHL‐knockdown HT1080 cells after treatment with GDAz‐3 and 8e for 24 h. D) Western blot analysis of GPX4 in 786‐O cells after treatment with GDAz‐3 and 8e for 24 h. E) Chemical structures of BRD4‐targeting HSP70‐PROTAC GDAz‐15–17. F) Western blot analysis of their protein degradation activity of BRD4 in the MOLT‐4 cell line after treatment for 48 h.

Article Snippet: The proteomics data were generated by Shanghai Bioprofile Co., Ltd. After treatment with compound GDAz‐3 (2 μM) or DMSO for 18 h, the samples were collected.

Techniques: Western Blot, Knockdown, Activity Assay