Biological Activity

Bioactivity / Mechanism

In G-415 and TGBC-2TKB gallbladder cancer cells, WYE-354 (1 μM, 18 h) reduced phosphorylation of P70S6K and 4E-BP1. Phospho-eIF4E and the corresponding total proteins showed no significant changes. These cellular readouts support inhibition of downstream mTOR signaling under the tested conditions. [1]

Literature-reported observations and conditions; not Adooq-tested protocols or general treatment recommendations.

Target & Activity

Target	Activity	Value
mTOR	IC50	5nM
PI3Kα	IC50	1.89μM
PI3Kγ	IC50	7.37μM
GCK	IC50	>50μM
Hck	IC50	>50μM
IKKα	IC50	>50μM
Lyn	IC50	>50μM
Met	IC50	>50μM
MK2	IC50	>50μM
p38α	IC50	>50μM
PDGFRα	IC50	>50μM
PKA	IC50	>50μM
PKBα	IC50	>50μM
PKCα	IC50	>50μM
PKCβ	IC50	>50μM
ROCK1	IC50	>50μM
RSK1	IC50	>50μM
Src	IC50	>50μM
VEGFR2	IC50	>50μM
Abl1	IC50	>50μM
AuroraB	IC50	>50μM
CDK1/CyclinB	IC50	>50μM
CDK2/CyclinA	IC50	>50μM
Chk1	IC50	>50μM
ERK2	IC50	>50μM
Fyn	IC50	>50μM

26 targets · Scroll to view more

Cellular Effect

Cell Line	Activity	Value	Description	Reference
HEK293	IC50	4.3 nM	Inhibition of flag-tagged truncated human mTOR (1360-2549) expressed in HEK293 cells by DELFIA assay	[PMID: 19645448]
LNCaP	IC50	355 nM	Cytotoxicity against human LNCAP cells after 3 days by MTS assay	[PMID: 19645448]

In Vitro Studies

WYE-354 sensitivity depended on the cellular model: ABCB1-overexpressing KB-C2 and HEK293/ABCB1 cells had higher viability IC50 values than matched comparator cells. Verapamil (5 μM) partially increased their sensitivity, indicating a limitation relevant to interpreting WYE-354 cell-response experiments. [2]

Downstream mTOR signaling in gallbladder cancer cells

Cell-based study · Human G-415 and TGBC-2TKB gallbladder cancer cells [1]

WYE-354 reduced phosphorylation of P70S6K and 4E-BP1. Phospho-eIF4E and the corresponding total protein signals showed no significant changes under these conditions. This is a cellular pathway readout, without a direct binding or ATP-competition measurement.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
P70S6K phosphorylation at Thr389	Qualitative Western blot Reduced relative to vehicle (qualitative blot)	[1]
4E-BP1 phosphorylation at Thr37/46	Qualitative Western blot Reduced relative to vehicle (qualitative blot)	[1]
Phospho-eIF4E (Ser209) and total P70S6K, 4E-BP1 and eIF4E	Qualitative Western blot No significant changes reported	[1]

Literature-Reported Treatment Conditions

Treatment concentration: 1 μM
Exposure duration: 18 h
Method: Cell treatment followed by Western blotting; β-actin loading control and total-protein comparisons.
Vehicle: 0.01% dimethylsulfoxide vehicle control
Reported limits: No numerical inhibition percentage, replicate count for the blot, salt/solvate specification or lot analysis supplied for this experiment.

Methods and source locations

Compound reported in the paper: WYE-354

Findings concern the reported WYE-354 material; no current Adooq SKU or lot validation.

Figure 1B; Results: WYE-354 reduces cell viability and phosphorylation; Methods: mTOR inhibitors, Antibodies, Western blot analysis

P70S6K phosphorylation at Thr389: Western blot; β-actin loading control — Figure 1B; Results: WYE-354 reduces cell viability and phosphorylation; Methods: mTOR inhibitors, Antibodies, Western blot analysis

4E-BP1 phosphorylation at Thr37/46: Western blot; β-actin loading control — Figure 1B; Results: WYE-354 reduces cell viability and phosphorylation; Methods: mTOR inhibitors, Antibodies, Western blot analysis

Phospho-eIF4E (Ser209) and total P70S6K, 4E-BP1 and eIF4E: Western blot; β-actin loading control — Figure 1B; Results: WYE-354 reduces cell viability and phosphorylation; Methods: mTOR inhibitors, Antibodies, Western blot analysis

ABCB1 expression and WYE-354 cellular sensitivity

Cell-based study · KB-3-1 and KB-C2 cancer cells; HEK293/pcDNA3.1 empty-vector and HEK293/ABCB1 comparator cells [2]

ABCB1-overexpressing models had higher WYE-354 viability IC50 values than their matched comparators. Verapamil reduced IC50 values in overexpressing cells but did not fully restore comparator sensitivity. These cellular IC50 values are not mTOR enzyme potency measurements.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
KB-3-1 viability after 72 h	Cellular IC50 WYE-354 alone: 1.410 ± 0.481 μM; + 5 μM verapamil: 2.135 ± 0.522 μM; mean ± SD of three independent experiments	[2]
KB-C2 viability after 72 h	Cellular IC50 WYE-354 alone: 9.937 ± 0.625 μM; + 5 μM verapamil: 3.788 ± 0.604 μM; mean ± SD of three independent experiments; combination versus control p < 0.05	[2]
HEK293/pcDNA3.1 viability after 72 h	Cellular IC50 WYE-354 alone: 2.319 ± 0.208 μM; + 5 μM verapamil: 2.764 ± 0.179 μM; mean ± SD of three independent experiments	[2]
HEK293/ABCB1 viability after 72 h	Cellular IC50 WYE-354 alone: 13.84 ± 1.465 μM; + 5 μM verapamil: 7.372 ± 0.165 μM; mean ± SD of three independent experiments; combination versus control p < 0.05	[2]

Literature-Reported Treatment Conditions

Treatment concentration: WYE-354 dose–response; plotted series 0–30 μM; comparator combination with 5 μM verapamil
Exposure duration: 72 h
Method: MTT cell viability assay; single-agent and verapamil-combination comparisons
Vehicle: DMSO control; final vehicle fraction not specified in the adopted assay section
Co-treatment: Separate WYE-354-alone and WYE-354 plus 5 μM verapamil conditions
Reported limits: No salt/solvate or lot specification. The Table 1 combination preincubation timing is not separately explicit; no timing is inferred from other combination assays.

Methods and source locations

Compound reported in the paper: WYE-354

Compound-level results using another supplier; no evidence of Adooq lot validation.

Results 2.1–2.2; Table 1 and footnotes; Figures 1B–C and 2; Methods 4.1, 4.3 and 4.9

KB-3-1 viability after 72 h: MTT assay; Table 1 — Table 1 including footnotes; Methods 4.3 and 4.9

KB-C2 viability after 72 h: MTT assay; Table 1 — Table 1 including footnotes; Methods 4.3 and 4.9

HEK293/pcDNA3.1 viability after 72 h: MTT assay; Table 1 — Table 1 including footnotes; Methods 4.3 and 4.9

HEK293/ABCB1 viability after 72 h: MTT assay; Table 1 — Table 1 including footnotes; Methods 4.3 and 4.9

In Vivo Studies

In NOD-SCID mice bearing G-415 or TGBC-2TKB gallbladder cancer xenografts, WYE-354 (50 mg/kg intraperitoneally, once daily for 5 days) reduced day-30 tumor volume by 68.6% and 52.4%, respectively, versus vehicle. TGBC-2TKB tumor-weight reduction was not significant. Terminal signaling inhibition was partial, and Ki67 decreased significantly only in TGBC-2TKB tumors. [1]

WYE-354 in two gallbladder cancer xenograft models

Animal study · 8–12-week-old NOD-SCID mice bearing subcutaneous G-415 or TGBC-2TKB tumors [1]

At day 30, WYE-354-treated tumors had smaller volumes in both models. Tumor-weight reduction was significant in G-415 but not TGBC-2TKB. Ki67 declined significantly only in TGBC-2TKB; G-415 showed a nonsignificant increase. Terminal P70S6K phosphorylation was only partly reduced, with marginal decreases in half of analyzed WYE-354 tumors.

Measured endpoints — not treatment concentrations
Endpoint	Reported result	Reference
G-415 tumor volume at day 30 versus matched control	Reduction 68.6%; P < 0.01; n = 5 per group; plotted data mean ± SD	[1]
G-415 terminal tumor weight versus control	Reduction 82.9%; P < 0.01; n = 5 per group; plotted data mean ± SD	[1]
TGBC-2TKB tumor volume at day 30 versus matched control	Reduction 52.4%; P < 0.01; n = 4 per group; plotted data mean ± SD	[1]
TGBC-2TKB terminal tumor weight versus control	Reduction 45.5%; not significant; n = 4 per group; plotted data mean ± SD	[1]
Terminal tumor Ki67 proliferative index	Change versus control TGBC-2TKB: 23.2% decrease, P < 0.01; G-415: higher Ki67, not significant; plotted data mean ± SD	[1]
Terminal tumor phospho-P70S6K	Qualitative Western blot Partial inhibition; only half of analyzed WYE-354 tumors showed a significant but marginal decrease, as described by authors	[1]

Conditions shared by the study groups

Route: Intraperitoneal
Frequency: Once daily
Treatment course: 5 days; followed to day 30
Vehicle: 5% ethanol, 5% Tween 80, 5% PEG 400

Study groups and reported outcomes
Group / dose	Comparator	Observed result	Reference
G-415 xenografts: WYE-354 versus vehicle 50 mg/kg WYE-354	Matched vehicle-treated xenografts, n = 5	At day 30, tumor volume reduced 68.6% (P < 0.01) and tumor weight reduced 82.9% (P < 0.01) versus control. Ki67 increased nonsignificantly.	[1]
TGBC-2TKB xenografts: WYE-354 versus vehicle 50 mg/kg WYE-354	Matched vehicle-treated xenografts, n = 4	At day 30, tumor volume reduced 52.4% (P < 0.01) and tumor weight reduced 45.5% (not significant) versus control. Ki67 decreased 23.2%, P < 0.01.	[1]

G-415 xenografts: WYE-354 versus vehicle: Model: NOD-SCID mice; G-415 subcutaneous xenografts; n = 5 per group (Figure 4); Reported limits: Sex not specified; animals sacrificed at approximately 1500 mm³ tumor volume or day 30 per caption.

TGBC-2TKB xenografts: WYE-354 versus vehicle: Model: NOD-SCID mice; TGBC-2TKB subcutaneous xenografts; n = 4 per group (Figure 4); Reported limits: Sex not specified; animals sacrificed at approximately 1500 mm³ tumor volume or day 30 per caption.

Methods and source locations

Compound reported in the paper: WYE-354

Findings concern the reported WYE-354 material; no current Adooq SKU or lot validation.

Figure 4A–E and caption; Results: Rapamycin and WYE-354 inhibit tumor growth; Methods: mTOR inhibitors, In vivo studies on subcutaneous tumors, Immunohistochemical staining, Statistical analysis

G-415 tumor volume at day 30 versus matched control: Caliper volume = 0.52 × width² × length; reported reduction in Results — Figure 4A; Results: Rapamycin and WYE-354 inhibit tumor growth

G-415 terminal tumor weight versus control: Terminal tumor weighing — Figure 4A; Results: Rapamycin and WYE-354 inhibit tumor growth

TGBC-2TKB tumor volume at day 30 versus matched control: Caliper volume = 0.52 × width² × length; reported reduction in Results — Figure 4B; Results: Rapamycin and WYE-354 inhibit tumor growth

TGBC-2TKB terminal tumor weight versus control: Terminal tumor weighing — Figure 4B; Results: Rapamycin and WYE-354 inhibit tumor growth

Terminal tumor Ki67 proliferative index: Immunohistochemistry; ImmunoRatio analysis; unpaired Mann–Whitney test — Figure 4E; Results terminal Ki67 paragraph; Methods: Immunohistochemical staining

Terminal tumor phospho-P70S6K: Tumor Western blots compared with total P70S6K and β-actin — Figure 4D; Results paragraph on terminal mTOR signaling; Methods: Western blot analysis

G-415 xenografts: WYE-354 versus vehicle · Figure 4A–E and caption; Results: Rapamycin and WYE-354 inhibit tumor growth; Methods: mTOR inhibitors, In vivo studies on subcutaneous tumors, Immunohistochemical staining, Statistical analysis

TGBC-2TKB xenografts: WYE-354 versus vehicle · Figure 4A–E and caption; Results: Rapamycin and WYE-354 inhibit tumor growth; Methods: mTOR inhibitors, In vivo studies on subcutaneous tumors, Immunohistochemical staining, Statistical analysis

Specifications

Manufacturer	Adooq Bioscience
Actions	Inhibitor
CAS Number	1062169-56-5
Applications	mTOR Inhibitor
Formula	C₂₄H₂₉N₇O₅
Molecular Weight	495.5
SMILES	COC(=O)NC1=CC=C(C=C1)C2=NC3=C(C=NN3C4CCN(CC4)C(=O)OC)C(=N2)N5CCOCC5

Storage & Solubility

In Vitro Solubility

Reported Solubility

Solvent	Reported Solubility
DMSO	95 mg/mL (191.71 mM)
Water	Insoluble
Ethanol	Insoluble

Preparing Stock Solutions

Stock preparation is unavailable for this product molecular weight.

Calculated stock concentrations do not establish solvent solubility. Refer to the reported solubility data for experimental guidance.

In Vivo Preparation

Concentration: 4 mg/mL
Vehicle: 4% DMSO+30% PEG 300+5% Tween 80+ddH2O

Storage

Store powder at -20°C, keep desiccated.
In powder form, the chemical is stable for 36 months.
In solution, store at -20°C and use within 1 month to prevent loss of potency. Aliquot to avoid multiple freeze/thaw cycles.

Research References

[1] Helga Weber, Pamela Leal, Stefan Stein, et al. Rapamycin and WYE-354 suppress human gallbladder cancer xenografts in mice Oncotarget (2015). PMID: 26397134 DOI: 10.18632/oncotarget.5047 Source material
[2] Jingqiu Wang, Dong-Hua Yang, Yuqi Yang, et al. Overexpression of ABCB1 Transporter Confers Resistance to mTOR Inhibitor WYE-354 in Cancer Cells International Journal of Molecular Sciences (2020). PMID: 32092870 DOI: 10.3390/ijms21041387 Source material

Publications

1 publication citing this product

H Weber ,etc. Rapamycin and WYE-354 suppress human gallbladder cancer xenografts in mice. Oncotarget, 2015, 6(31), 31877.

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WYE-354

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WYE-354 5 mg

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Biological Activity

Bioactivity / Mechanism

Target & Activity

Cellular Effect

In Vitro Studies

Downstream mTOR signaling in gallbladder cancer cells

Literature-Reported Treatment Conditions

ABCB1 expression and WYE-354 cellular sensitivity

Literature-Reported Treatment Conditions

In Vivo Studies

WYE-354 in two gallbladder cancer xenograft models

Conditions shared by the study groups

Specifications

Storage & Solubility

In Vitro Solubility

Reported Solubility

Preparing Stock Solutions

In Vivo Preparation

Storage

Research References

Publications

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