Home Apoptosis Bcl-2 inhibitor - Autophagy activator - Apoptosis related activator (R)-(-)-Gossypol (AT-101) acetic acid

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(R)-(-)-Gossypol (AT-101) acetic acid Bcl-2 inhibitor

Cat.No.S2812

(R)-(-)-Gossypol (AT-101) acetic acid, the R-(-) enantiomer of Gossypol acetic acid, binds with Bcl-2, Bcl-xL and Mcl-1 with Ki of 0.32 μM, 0.48 μM and 0.18 μM in cell-free assays; does not inhibit BIR3 domain and BID. AT-101 simultaneously triggers apoptosis and a cytoprotective type of autophagy. Phase 2.
(R)-(-)-Gossypol (AT-101) acetic acid Bcl-2 inhibitor Chemical Structure

Chemical Structure

Molecular Weight: 578.61

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Chemical Information, Storage & Stability

Molecular Weight 578.61 Formula

C30H30O8.C2H4O2

 Storage (From the date of receipt)
CAS No. 866541-93-7 Download SDF Storage of Stock Solutions

Solubility

In vitro
Batch:

DMSO : 100 mg/mL (172.82 mM)
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : 100 mg/mL

Water : Insoluble

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In vivo
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Mechanism of Action

Targets/IC50/Ki
Mcl-1 [1]
(Cell-free assay)
0.18 μM(Ki)
Bcl-2 [1]
(Cell-free assay)
0.32 μM(Ki)
Bcl-xL [1]
(Cell-free assay)
0.48 μM(Ki)
In vitro
AT-101 inhibits a panel of different lymphoproliferative malignancies with IC50 ranged from 1.2 μM to 7.4 μM. AT-101 (10 μM) disrupts the Δψm in a concentration- and time-dependent manner in a diffuse large B-cell and in mantle cell lymphoma lines. AT-101 (1 μM or 2 μM) combined with carfilzomib (6 nM or 10 nM) induces apoptosis in HBL-2 and Granta cell lines. [2] AT-101 (20 μM for 24 hours) results in a median 72% apoptosis and down-regulation of Mcl-1 in CLL lymphocytes in both suspension culture as well as stromal coculture. Stromal cells express undetectable levels of antiapoptotic but high levels of activated ERK and AKT proteins and has low or no apoptosis with AT-101. [3] AT-101 induces apoptosis in a time- and dose-dependent fashion, with ED50 values of 1.9 mM and 2.4 mM in Jurkat T and U937 cells, respectively. AT-101 (10 μM) combined with radiation (32 Gy) induces more apoptosis than radiation alone and exceeds the sum of the effects caused by the single agent treatments. AT-101 activates SAPK/JNK in a dose- and time-dependent manner. [4] AT-101 (10 µM) induces apoptosis through activation of caspase-9, -3, and -7 in VCaP Cells. AT-101 (10 µM) decreases Bcl-2 and Mcl-1 expression in VCaP cells. [5] AT-101 (< 20 μM) is able to inhibit the growth of multiple myeloma cells despite the stimulatory growth effects provided by stromal cells in the bone marrow milieu. AT-101 (10 μM) induces apoptosis in multiple myeloma cells via the activation of caspases 3, caspases 9 and PARP. AT-101 (10 μM) promotes apoptosis in multiple myeloma cells by disrupting the Bax/Bcl-2 ratio and the mitochondrial membrane potential. [6]
Kinase Assay
Fluorescence-Polarization-Based Binding Assay
For competitive binding experiments, Bcl-2 protein (40 nM) and FAM-Bid peptide (2.5 nM) are preincubated in the assay buffer (100 mM potassium phosphate, pH 7.5; 100 μg/mL bovine gamma globulin; 0.02% sodium azide, 5 μL of a solution in DMSO of AT101 is added to the Bcl-2/FAM-Bid solution in Dynex 96-well, black, round-bottom plates to produce a final volume of 125 μL. For each experiment, a control containing Bcl-2 and Flu-Bid peptide (equivalent to 0% inhibition), and another control containing only FAM-Bid, are included on eachassay plate. After 4 hours incubation, the polarization values in milipolarization units (mP) weremeasured at an excitation wavelength at 485 nm and an emission wavelength at 530 nm using the Ultra plate reader. IC50,the inhibitor concentration at which 50% of bound peptide is displaced, is determined from the plot using nonlinear leastsquares analysis and curve fitting performed using GraphPad Prizm 4 software. The unlabeled Bid BH3 peptide is used as the positive control. PF for Bcl-xL protein, Bak BH3 peptide labeled with 6-carboxyfluorescein succinimidyl ester (FAM-Bak) instead of the FAM-Bim to maximize the signal. It is determined that FAM-Bak has a Kd of 6 nM to Bcl-xL protein. The competitive binding assay for Bcl-xL is same as that for Bcl-2 with the following exceptions. 30 nM of Bcl-xL protein and 2.5 nM of FAM-Bak peptide in the following assay buffer: 50 mM Tris-Bis, pH 7.4 and 0.01% bovine gamma globulin. PF for Mcl-1 protein, FAM-Bid peptide and human Mcl-1 protein are used. It is determined that FAM-Bid peptide binds to human Mcl-1 protein with a Kd of 1.71 nM. The competitive binding assays for Mcl-1 are performed in the same manner as that for Bcl-2 with the following exceptions. 5 nM Mcl-1 and 1 nM Flu-Bid peptide in the following assay buffer: 25 mM Tris, pH 8.0; 150 mM NaCl and 0.05% Pluronic acid
In vivo
AT-101 is still detectable in plasma with average concentrations of 0.49 μM for the 35 mg/kg group and 0.39 μM for the 200 mg/kg group in SCID beige mice bearing RL-DLBCL xenograft. AT-101 peak plasma concentration is observed after 30 minutes of administration of the drug in both the dose levels, with the 200 mg/kg group showing a plasma average concentration almost 4 times greater than the 35 mg/kg group (7.88 μM and 27.78 μM respectively) in SCID beige mice. AT-101 (25 mg/kg to 100 mg/kg, orally) indefinitely results in earlier onset of weight loss equivalent to more than 10% of the pretreatment weight and death in SCID beige mice. AT-101 (35 mg/kg, orally per day for 10 days) plus intraperitoneal cyclophosphamide (Cy) and intraperitoneal rituximab (R) show significantitumor volume control compared to any other treatment group. [2] AT-101 (15 mg/kg, p.o., 5 days/week) as a single agent in intact mice significantly reduces the development of VCaP tumor growth compared to untreated tumors at weeks 2 to 6. AT-101 in combination with surgical castration delays the onset of androgen-independent VCaP tumor growth compared to castration-only or AT-101-only groups in mice. [5]
References
  • [4] https://pubmed.ncbi.nlm.nih.gov/19852810/
  • [5] https://pubmed.ncbi.nlm.nih.gov/18084610/
  • [6] https://pubmed.ncbi.nlm.nih.gov/18346839/

Applications

Methods Biomarkers Images PMID
Growth inhibition assay Cell viability S2812-viability1 24824755
Western blot BNIP3 Smac / Cyt C p53 / Cox IV p-AKT / AKT APE1 / Bcl-2 / p53 / p-p53 / NF-κB S2812-WB1 24824755

Clinical Trial Information

(data from https://clinicaltrials.gov, updated on 2024-05-22)

NCT Number Recruitment Conditions Sponsor/Collaborators Start Date Phases
NCT05338931 Recruiting
B-cell Non Hodgkin Lymphoma
AbClon
 March 15 2022 Phase 1|Phase 2
NCT00934076 Withdrawn
Carcinoma Non Small Cell Lung
University of Alabama at Birmingham|Ascenta Therapeutics
 February 2010 Phase 1
NCT00561197 Terminated
Locally Advanced Esophageal or GE Junction Cancer
Ascenta Therapeutics
 August 2007 Phase 1|Phase 2

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Frequently Asked Questions

Question 1:
Is it S2812 (-)-gossypol or another enantiomer?

Answer:
It is the R-(–)-gossypol acetic acid.

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