17-DMAG reduced primary CLL-cell viability with increasing concentration and exposure time. Normal T and NK cells showed no significant toxicity in the same study, while normal B cells showed some toxicity. This selectivity was observed in isolated-cell assays. [1]
In primary prostate cancer-associated fibroblasts, 17-DMAG reduced collagen-gel contraction at 6–60 nM over 48 hours and reduced scratch closure at 30 nM over 24 hours. These functional endpoints were measured in separate assay designs. [2]
Sensitivity to 17-DMAG can change with acquired resistance. In ALK-rearranged lung cancer cell models, resistant derivatives showed increased P-glycoprotein expression; P-glycoprotein silencing or verapamil restored responsiveness. In the parental models, dicumarol did not change sensitivity to 17-DMAG. [3]
CLL-cell viability and normal-lymphocyte comparison
Cell-based study · Primary CD19-selected CLL B cells; normal donor B, T and NK cells. [1]
17-DMAG reduced CLL-cell viability with increasing concentration and exposure time. Normal T and NK cells did not show significant toxicity under the tested conditions; normal B cells showed some toxicity.
Literature-Reported Treatment Conditions
- Treatment concentration
- CLL annexin V/PI: 10, 50, 100 and 500 nM; normal lymphocytes: 100 nM, 500 nM and 1 μM.
- Exposure duration
- 24 or 48 hours
- Method
- Annexin V/propidium iodide flow cytometry; double-negative cells counted as live; 10,000 cells/sample.
- Co-treatment
- 17-DMAG alone unless a separately identified comparator or mechanistic rescue condition is specified.
- Reported limits
- Vehicle composition for these cellular assays is not specified. Error-bar definition in Figure 1 is not stated. Normal-donor and patient sample counts differ.
Methods and source locations
Compound reported in the paper: 17-DMAG; Compound label in the paper: 17-DMAG
Findings concern literature 17-DMAG and do not establish hydrochloride-specific potency, formulation equivalence or Adooq product validation.
Methods: Patients, cell separation, culture conditions, and reagents; Viability assays; Statistical analysis; Results: selective apoptosis; Figure 1A (left) and 1B
CLL-cell viability: Annexin V/PI; linear mixed-effects analysis described in Methods. — Results: 17-DMAG promotes selective apoptosis in CLL cells; Figure 1A left and caption; Statistical analysis
Viability of normal lymphocytes: Annexin V/PI at 24 and 48 hours. No percent viability estimated from bars. — Figure 1B and caption; Results: 17-DMAG promotes selective apoptosis in CLL cells
IKK depletion and NF-κB signaling in primary CLL cells
Cell-based study · Primary CLL patient cells. [1]
17-DMAG exposure decreased IKKα/IKKβ and AKT protein levels and reduced NF-κB signaling. At 1 μM for 24 hours, nuclear NF-κB DNA binding and MCL1/BCL2 expression decreased. These are cellular pathway readouts, not direct IKK enzyme inhibition measurements.
Literature-Reported Treatment Conditions
- Treatment concentration
- Figure 3: 10 nM, 100 nM and 1 μM; Figure 4: 1 μM.
- Exposure duration
- 24 hours
- Method
- Immunoblotting, nuclear-extract electrophoretic mobility shift assay and real-time PCR.
- Co-treatment
- 17-DMAG alone unless a separately identified comparator or mechanistic rescue condition is specified.
- Reported limits
- Cellular vehicle composition is not specified. No binding constant or enzyme IC50 is established by these experiments.
Methods and source locations
Compound reported in the paper: 17-DMAG; Compound label in the paper: 17-DMAG
Findings concern literature 17-DMAG and do not establish hydrochloride-specific potency, formulation equivalence or Adooq product validation.
Methods: Immunoblot analysis; Electrophoretic mobility shift assay; Real-time PCR; Statistical analysis; Figures 3 and 4A–D; corresponding Results
IKKα, IKKβ and AKT protein abundance: Immunoblot with actin loading controls. — Figure 3 image and caption; Results: 17-DMAG leads to degradation of IKK-α and β
NF-κB nuclear signaling and DNA binding: Immunoblot and EMSA after 1 μM 17-DMAG for 24 hours. TNF-α and Bay-11 are separate assay comparators, not combination treatment. — Figure 4A–B and caption; corresponding Results
MCL1 and BCL2 transcript abundance: Real-time PCR after 1 μM for 24 hours; Figure 4C normalized to 18S; points are individual patients and bar is the average. — Figure 4C and caption; Results: 17-DMAG depletion of IKK results in inhibition of NF-κB activation and signaling
Collagen-gel contraction by prostate cancer-associated fibroblasts
Cell-based study · Primary prostate cancer-associated fibroblasts (CAF), three patient-derived lines. [2]
17-DMAG reduced collagen-gel contraction at 6, 20 and 60 nM after 48 hours. This endpoint measures gel contraction; it does not by itself establish a direct molecular target or exclude cellular toxicity.
Literature-Reported Treatment Conditions
- Treatment concentration
- 6, 20 and 60 nM; 0 nM vehicle control.
- Exposure duration
- 48 hours
- Method
- CAF embedded at 25,000 cells/mL in 1 mg/mL collagen; 0.5 mL gels in 24-well plates with DMEM/5% FCS; dislodged gels imaged for area-based contraction.
- Co-treatment
- 17-DMAG alone unless a separately identified comparator or mechanistic rescue condition is specified.
- Reported limits
- Vehicle chemical identity is not given. The associated MTS endpoint is excluded because its reagent incubation is reported as 2 hours in Methods 4.3 and 3 hours in Figure 3 legend.
Methods and source locations
Compound reported in the paper: 17-DMAG; Compound label in the paper: 17-DMAG
Findings concern literature 17-DMAG and do not establish hydrochloride-specific potency, formulation equivalence or Adooq product validation.
Results 2.5; Methods 4.1–4.3 and 4.7; Figure 3C and caption
Collagen-gel contraction: Image-based collagen-gel contraction at 48 hours. No bar heights digitized. — Figure 3C and caption; Results 2.5
Scratch closure by prostate cancer-associated fibroblasts
Cell-based study · Primary prostate cancer-associated fibroblasts from five patients. [2]
17-DMAG at 30 nM reduced scratch closure over 24 hours.
Literature-Reported Treatment Conditions
- Treatment concentration
- 30 nM
- Exposure duration
- 24 hours
- Method
- Confluent CAF in 24-well plates scratched with a pipette tip, washed with PBS and incubated in DMEM/1% FCS; images at 0 and 24 hours analyzed with ImageJ.
- Co-treatment
- 17-DMAG alone unless a separately identified comparator or mechanistic rescue condition is specified.
- Reported limits
- Vehicle chemical identity is not given. Scratch closure is a composite assay and does not isolate migration from every other contributor.
Methods and source locations
Compound reported in the paper: 17-DMAG; Compound label in the paper: 17-DMAG
Findings concern literature 17-DMAG and do not establish hydrochloride-specific potency, formulation equivalence or Adooq product validation.
Results 2.6; Methods 4.3 and 4.7; Figure 3F and caption
Scratch wound closure: Scratch assay at 24 hours; quadruplicate measurements per CAF population described in Methods. — Results 2.6; Figure 3F and caption; Methods 4.3
Acquired 17-DMAG resistance and P-glycoprotein rescue
Cell-based study · ALK-rearranged H3122 and H2228 cells; resistant derivatives H3122/DR-1, H3122/DR-2 and H2228/DR. [3]
Chronic selection produced 17-DMAG-resistant derivatives with increased P-glycoprotein expression and rhodamine efflux. P-glycoprotein silencing or verapamil restored drug responsiveness. Dicumarol did not change parental-cell sensitivity, arguing against NQO1 inhibition as the principal explanation in these models.
Literature-Reported Treatment Conditions
- Treatment concentration
- 17-DMAG response curves: 0.01–10 μM in Figure 1; 0.001–10 μM in Figure 4E. Mechanistic conditions: 5 μM verapamil, 100 nM P-gp siRNA, or 20 μM dicumarol.
- Exposure duration
- 72-hour drug-response assays; resistant lines selected over 6 months and kept drug-free for >1 week before experiments.
- Method
- MTT response assays; eight replicate wells per assay, repeated at least three times. Protein/RNA analyses, siRNA and pharmacological perturbation, and rhodamine-123 efflux assay.
- Co-treatment
- Response curves include 17-DMAG alone; separate rescue conditions use 5 μM verapamil or 100 nM P-gp siRNA. NQO1 test combines 17-DMAG with 20 μM dicumarol. AUY922 and ALK inhibitors were separate comparators.
- Reported limits
- Vehicle composition and verapamil pretreatment interval are not specified. IC50 uncertainties are reproduced exactly as printed; unusually large SD values are not corrected or re-estimated.
Methods and source locations
Compound reported in the paper: 17-DMAG; Compound label in the paper: 17-DMAG
Findings concern literature 17-DMAG and do not establish hydrochloride-specific potency, formulation equivalence or Adooq product validation.
Methods: cell culture/reagents, resistance establishment, MTT, rhodamine efflux and siRNA; Results; Figures 1, 3E and 4A–F
MTT growth-inhibition response to 17-DMAG: 72-hour MTT; at least three independent experiments. Values transcribed from Figure 1 table, not curves. — Figure 1 embedded table and caption; Methods: MTT assay
Responsiveness of resistant cells to 17-DMAG: 72-hour MTT after siRNA transfection or verapamil pretreatment; corresponding P-gp immunoblots. — Figure 4B, D–E and caption; Results: The induction of P-gp leads to 17-DMAG resistance
17-DMAG response with NQO1 inhibitor: MTT combination experiment. — Figure 3E and caption; Results: NQO1 expression is not associated with acquired resistance to 17-DMAG
