Cat. No.: HY-187731

BXL0124

BXL0124 is an orally effective CD44 inhibitor and Notch signaling pathway inhibitor. BXL0124 has a vitamin D receptor-dependent mechanism and can downregulate the expression of CD44, Notch1/2/3, HES1, OCT4, LAMA5, JAG1, JAG2, NF-κB and DLL1. BXL0124 inhibits c-Myc expression and the levels of phosphorylated ERK, AKT, ErbB2, reduces the level of activated Notch1 receptor and its nuclear localization, decreases the mRNA and protein levels of Jagged-1 and Jagged-2, and inhibits the STAT3 signaling pathway by reducing the formation of the CD44-STAT3-JAK2 complex, while also inhibiting the transcriptional activity of the CD44 promoter in a p53-dependent manner. BXL0124 can induce myoepithelial differentiation and inhibit the self-renewal of cancer stem cell-like cells, cancer cell proliferation, invasion, and growth. BXL0124 can be used in research related to triple-negative breast cancer, basal-like breast cancer, ErbB2-overexpressing mammary tumorigenesis, and breast cancer.

For research use only. We do not sell to patients.

CAS No.: 685141-23-5
Formula: C₃₂H₄₄F₆O₄
Molecular Weight:606.68

Storage:

Please store the product under the recommended conditions in the Certificate of Analysis.

BXL0124 Chemical Structure

CAS No.: 685141-23-5

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Use of MCE Products

Strong Fluorescence Signal / Multi-Disciplinary Applications / High Fluorescence Specificity

Biological Activity
- Description
- IC₅₀ & Target
- In Vitro
- In Vivo
Chemical Information
Purity & Documentation
References
Calculators
Help & FAQs
Classifications
Bioz

Biological Activity

Description

IC₅₀ & Target

^[1]

STAT3

All NF-κB Isoforms :

All STAT Isoforms :

All JAK Isoforms :

In Vitro

Anti-cancerCell Cycle Regulation

Anti-cancer

BXL0124 (1-100 nM; 5 days) potently reduces mammosphere formation efficiency and self-renewal capacity in SUM159 triple-negative breast cancer cells^[1].
BXL0124 (10 nM; 5 days) downregulates the expression of pluripotency and cancer stem cell markers in SUM159 triple-negative breast cancer mammospheres^[1].
BXL0124 (10 nM; 5 days) inhibits the expression of Notch signaling pathway molecules in SUM159 triple-negative breast cancer mammospheres^[1].
BXL0124 (10 nM; 5 days) can modulate the expression of mammary epithelial differentiation markers in SUM159 triple-negative breast cancer mammospheres^[1].
BXL0124 (10 nM; 72 h) inhibits the proliferation of CD44+/CD24−, CD44+/CD24_low, and CD44+/CD24_high MCF10DCIS basal-like breast cancer cells^[2].
BXL0124 (10 nM; 24 h) reduces the protein levels of c-Notch1, Jagged-2, and c-Myc in CD44⁺/CD24⁻, CD44⁺/CD24^low, and CD44⁺/CD24^high MCF10DCIS basal-like breast cancer cells^[2].
BXL0124 (0.1-10 nM; 24 h) inhibits the Notch signaling pathway in MCF10DCIS basal-like breast cancer cells in a dose-dependent manner in vitro, reducing the levels of c-Notch1, Notch3, Jagged-1, Jagged-2, DLL1, and c-Myc proteins^[2].
BXL0124 (10 nM; 1-24 h) decreased the protein levels of c-Notch1, Jagged-1, and Jagged-2 in MCF10DCIS basal-like breast cancer cells in a time-dependent manner^[2].
BXL0124 (10 nM; 24 h) reduces nuclear levels of activated c-Notch1 and increases nuclear VDR levels in MCF10DCIS basal-like breast cancer cells^[2].
BXL0124 (10 nM; 24 h) inhibits the Notch1 signaling pathway in MCF10DCIS basal-like breast cancer cells in a VDR-dependent manner^[2].
BXL0124 (10 nM; 24 h) inhibits Notch1 signaling in MCF10DCIS basal-like breast cancer cells in a HES1-dependent manner^[2].
BXL0124 (10 nM; 1-24 h) increases HES1 mRNA levels in MCF10DCIS basal-like breast cancer cells, while decreasing the mRNA levels of Jagged-1, Jagged-2, DLL1, and c-Myc, but the mRNA levels of Notch receptors remain unchanged^[2].
BXL0124 (10 nM; 30 min-24 h) rapidly and persistently upregulates HES1 mRNA and protein levels in MCF10DCIS basal-like breast cancer cells^[2].
BXL0124 (administered continuously for 3 days) potently inhibits the proliferation of ErbB2-overexpressing E18-9A-42 breast tumor cells, with an IC₅₀ of 0.04 nM^[3].
BXL0124 (24 h incubation) reduces plasma membrane-localized phosphorylated AKT levels in E18-9A-42 ErbB2-overexpressing breast tumor cells^[3].
BXL0124 (0.01-100 nM; 72 h) potently inhibits the proliferation of MCF10DCIS basal-like human breast cancer cells^[4].
BXL0124 (0.01-100 nM; 24-72 h) inhibits the metabolic activity of MCF10DCIS basal-like human breast cancer cells^[4].
BXL0124 (1-10 nM; 48 h) inhibits the invasion of MCF10DCIS basal-like human breast cancer cells in a BME-coated invasion assay^[4].
BXL0124 (1-10 nM; 10 days) inhibits the invasive growth of MCF10DCIS basal-like human breast cancer cells in 3D Matrigel culture assays^[4].
BXL0124 (10 nM; 24-48 h) inhibits the mRNA expression of CD44, MMP-2, MMP-9, and uPA in MCF10DCIS basal-like human breast cancer cells^[4].
BXL0124 (0.1-10 nM; 24 h) dose-dependently inhibited the protein levels of CD44 and pSTAT3 in MCF10DCIS basal-like human breast cancer cells, without altering the protein levels of total STAT3, Akt, Erk, or NFκB^[4].
BXL0124 (10 nM; 24 h)-mediated downregulation of CD44 and pSTAT3 protein levels in MCF10DCIS basal-like human breast cancer cells is a VDR-dependent process^[4].
BXL0124 (10 nM; 24 h) reduces nuclear localization of phosphorylated STAT3 in MCF10DCIS basal-like human breast cancer cells^[4].
BXL0124 (0.1-10 nM; 24 h) inhibits STAT3 DNA-binding activity in MCF10DCIS basal-like human breast cancer cells in a dose-dependent manner^[4].
BXL0124 (10 nM; 24 h) reduces the formation of CD44-STAT3-JAK2 protein complexes in MCF10DCIS basal-like human breast cancer cells and decreases the levels of phosphorylated STAT3 in these complexes^[4].
BXL0124 (10 nM; 24 h) upregulates VDR protein levels and inhibits CD44 and pSTAT3 protein levels in MCF10CA1a and MDA-MB-468 basal-like human breast cancer cells^[4].
BXL0124 (0.01-10 nM; 1-72 h) potently inhibits CD44 expression through a VDR- and p53-dependent mechanism, reduces the CD44⁺/high/CD24⁻/low breast cancer stem cell subpopulation, and induces osteopontin mRNA expression in MCF10DCIS.com human breast cancer cells in vitro^[5].

Anti-cancer

Cell Cycle Regulation

BXL0124 (incubation for 24 h) downregulates the phosphorylation levels of ErbB2, ERK, and AKT in E18-9A-42 ErbB2-overexpressing breast tumor cells and decreases cyclin D1 expression, without affecting the total protein levels of ErbB2, ERK, or AKT^[3].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Cell Proliferation Assay^[2]

Cell Line:

CD44+/CD24−, CD44+/CD24low, and CD44+/CD24high subpopulations of MCF10DCIS basal-like breast cancer cells
Concentration:

10 nM
Incubation Time:

72 h
Result:

Reduced cell proliferation rates in all three sorted subpopulations to ~25% of control in CD44+/CD24− cells, ~28% of control in CD44+/CD24low cells, and ~20% of control in CD44+/CD24high cells.

Western Blot Analysis^[2]

Cell Line:

CD44+/CD24−, CD44+/CD24low, and CD44+/CD24high subpopulations of MCF10DCIS basal-like breast cancer cells
Concentration:

10 nM
Incubation Time:

24 h
Result:

Decreased protein levels of activated Notch1 (c-Notch1), Jagged-2, and c-Myc in all three sorted subpopulations.

Western Blot Analysis^[2]

Cell Line:

MCF10DCIS basal-like breast cancer cells
Concentration:

0.1-10 nM
Incubation Time:

24 h
Result:

Decreased protein levels of c-Notch1, total Notch3, Jagged-1, Jagged-2, DLL1, and c-Myc in a dose-dependent manner, with the greatest effects on c-Notch1 and Jagged-2.
Left total Notch1 and total Notch2 protein levels unaffected.
Showed stronger inhibitory activity than 1α,25(OH)₂D₃ at equivalent concentrations.

Western Blot Analysis^[2]

Cell Line:

MCF10DCIS basal-like breast cancer cells
Concentration:

10 nM
Incubation Time:

1-24 h
Result:

Markedly reduced c-Notch1 protein levels at 12 h and 24 h, while total Notch1 levels remained unchanged.
Reduced Jagged-1 protein levels at 4 h, 12 h, and 24 h.
Reduced Jagged-2 levels at 12 h and 24 h.

Western Blot Analysis^[2]

Cell Line:

MCF10DCIS basal-like breast cancer cells
Concentration:

10 nM
Incubation Time:

24 h
Result:

Had its ability to decrease c-Notch1 and Jagged-2 protein levels reduced by VDR knockdown, confirming inhibition of Notch1 activation is VDR-dependent.\nHad its ability to decrease c-Notch1 and Jagged-2 protein levels reduced by partial HES1 knockdown, indicating inhibition of Notch1 signaling is HES1-dependent.
Still increased HES1 protein levels despite knockdown.
Left VDR induction unaffected by HES1 knockdown.

Real Time qPCR^[2]

Cell Line:

MCF10DCIS basal-like breast cancer cells
Concentration:

10 nM
Incubation Time:

1-24 h
Result:

Did not affect mRNA levels of Notch1, Notch2, or Notch3.
Significantly decreased mRNA levels of Jagged-1, Jagged-2, and DLL1 at 8 h, 16 h, and 24 h.
Significantly decreased c-Myc mRNA levels at 16 h and 24 h.
Rapidly and significantly increased HES1 mRNA levels at all time points tested, including as early as 1 h.

Cell Proliferation Assay^[4]

Cell Line:

MCF10DCIS basal-like human breast cancer cells
Concentration:

0.01-100 nM
Incubation Time:

72 h
Result:

Potently inhibited MCF10DCIS cell proliferation, with greater potency than 1α,25(OH)₂D₃.
Caused a marked reduction in [³H]thymidine incorporation compared to untreated cells at 1, 10, and 100 nM.

Cell Viability Assay^[4]

Cell Line:

MCF10DCIS basal-like human breast cancer cells
Concentration:

0.01-100 nM
Incubation Time:

24-72 h
Result:

Inhibited MCF10DCIS cell metabolic activity in a time-dependent manner, with greater potency than 1α,25(OH)₂D₃.
Caused a significant reduction in absorbance at 560 nm compared to untreated cells at 10 and 100 nM at 72 h.

Cell Invasion Assay^[4]

Cell Line:

MCF10DCIS basal-like human breast cancer cells
Concentration:

1-10 nM
Incubation Time:

48 h
Result:

Significantly reduced the number of MCF10DCIS cells that penetrated the BME layer, with greater effectiveness than 1α,25(OH)₂D₃.
Caused a marked decrease in invaded cell counts compared to untreated cells at 10 nM.

Cell Invasion Assay^[4]

Cell Line:

MCF10DCIS basal-like human breast cancer cells
Concentration:

1-10 nM
Incubation Time:

10 days (medium replenished every 2 days)
Result:

Abolished the invasive outgrowth of MCF10DCIS cells observed in untreated control cultures.
Resulted in only non-invasive cell clusters, unlike control cultures which exhibited invasive protrusions.

Real Time qPCR^[4]

Cell Line:

MCF10DCIS basal-like human breast cancer cells
Concentration:

10 nM
Incubation Time:

24-48 h
Result:

Significantly decreased mRNA expression levels of CD44, MMP-2, MMP-9, and uPA at both 24 h and 48 h.
Did not significantly change mRNA expression of MMP-14.

Western Blot Analysis^[4]

Cell Line:

MCF10DCIS basal-like human breast cancer cells
Concentration:

0.1-10 nM (24 h dose-dependent study); 10 nM (6-24 h time-dependent study)
Incubation Time:

24 h (dose-dependent study); 6-24 h (time-dependent study)
Result:

Decreased protein levels of CD44 variant isoforms (CD44v, 100-250 kDa), CD44 standard isoform (CD44s, 85 kDa), and phosphorylated STAT3 (pSTAT3) in a dose-dependent manner at 24 h.
Left total STAT3, phosphorylated Akt (pAkt), total Akt, phosphorylated Erk (pErk), total Erk, phosphorylated NFκB (pNFκB), and total NFκB levels unchanged.
Decreased CD44s, CD44v, and pSTAT3 levels at 12 h and 24 h in a time-dependent study, while total STAT3 levels remained unchanged.

Western Blot Analysis^[4]

Cell Line:

MCF10DCIS basal-like human breast cancer cells
Concentration:

10 nM
Incubation Time:

24 h (after 72 h siRNA transfection)
Result:

Abolished the repression of CD44v, CD44s, and pSTAT3 protein levels in cells transfected with VDR siRNA, compared to cells transfected with non-targeting siRNA.

Immunofluorescence^[4]

Cell Line:

MCF10DCIS basal-like human breast cancer cells
Concentration:

10 nM
Incubation Time:

24 h
Result:

Reduced strong nuclear staining of pSTAT3 observed in untreated control cells, indicating decreased nuclear localization of activated STAT3.

Western Blot Analysis^[4]

Cell Line:

MCF10DCIS basal-like human breast cancer cells
Concentration:

10 nM
Incubation Time:

24 h
Result:

Decreased the amounts of CD44v, CD44s, and pSTAT3 in complexes immunoprecipitated with anti-STAT3 antibody, while total STAT3 levels remained unchanged.
Decreased the amount of JAK2 associated with STAT3, while Src association with STAT3 was unchanged.
Decreased the amounts of CD44v, CD44s, and pSTAT3 in complexes immunoprecipitated with anti-JAK2 antibody, while JAK2 levels remained unchanged.

Western Blot Analysis^[4]

Cell Line:

MCF10CA1a and MDA-MB-468 basal-like human breast cancer cells
Concentration:

10 nM
Incubation Time:

24 h
Result:

Increased vitamin D receptor (VDR) protein levels in both cell lines.
Decreased protein levels of CD44v, CD44s, and pSTAT3 in both cell lines.
Left total STAT3 protein levels unchanged in both cell lines.

Western Blot Analysis^[5]

Cell Line:

MCF10DCIS.com human breast cancer cells
Concentration:

0.01-10 nM (Western blot, 24 h); 10 nM (flow cytometry, fluorescence microscopy, VDR siRNA assay, time-course mRNA analysis, 24 h); 0.01-10 nM (dose-dependent mRNA analysis, 4 h); 0.1-10 nM (CD44 promoter assay, 24 h); 1 μM VDR siRNA (72 h, followed by 10 nM BXL0124 for 24 h)
Incubation Time:

24 h (Western blot, flow cytometry, fluorescence microscopy, VDR siRNA assay, CD44 promoter assay); 1, 4, 12, 24 h (time-course mRNA analysis); 4 h (dose-dependent mRNA analysis); 72 h (VDR siRNA incubation, followed by 24 h BXL0124 treatment)
Result:

Down-regulated CD44 standard isoform (CD44s, 85 kDa) and CD44 variant isoforms (CD44v, 100-250 kDa) protein expression in a dose-dependent manner, with greater potency than 1α,25-dihydroxyvitamin D₃.
Decreased the fraction of CD44⁺/high/CD24^-/low cells from 17.0% to 8.5%, and increased the fraction of CD44⁺/high/CD24⁺/high cells from 83.0% to 91.5%.
Reduced CD44 protein expression on the plasma membrane, with no change in subcellular localization.
Had its CD44 protein repression effect blocked by VDR siRNA knockdown, confirming VDR dependence.
Down-regulated CD44 mRNA expression with maximum inhibition at 4 h, and in a dose-dependent manner at 4 h, with greater potency than 1α,25-dihydroxyvitamin D₃.
Induced osteopontin mRNA expression starting at 4 h, with stronger induction than 1α,25-dihydroxyvitamin D₃ at 24 h, and in a dose-dependent manner at 4 h.
Repressed transactivation of the full-length CD44 promoter in a dose-dependent manner, with greater potency than 1α,25-dihydroxyvitamin D₃; this repression was abolished by p53-binding site mutation in the CD44 promoter, confirming p53 dependence.

In Vivo

Anti-cancerCell Cycle Regulation

Anti-cancer

Cell Cycle Regulation

BXL0124 (p.o.) delays MMTV-ErbB2/neu-induced mammary tumor formation by inhibiting ErbB2 signaling, with enhanced efficacy when combined with CDDO-IM and no hypercalcemic toxicity^[1].
BXL0124 (0.3 μg/kg BW; i.p.; three times weekly; 54 weeks) inhibits ErbB2-overexpressing mammary tumor growth by 57% (tumor volume) and 54% (tumor weight) in MMTV-ErbB2/neu transgenic mice via down-regulation of ErbB2/AKT/ERK signaling and cyclin D1 expression, without inducing hypercalcemic toxicity^[3].
BXL0124 (0.1 μg/kg; p.o.; 6 days a week; 5 weeks) suppresses MCF10DCIS.com xenograft breast tumor growth by 52% (tumor weight reduction) and represses CD44 and PCNA expression in nu/nu mice without hypercalcemic toxicity^[5].

Anti-cancer

BXL0124 inhibits growth of MCF10DCIS xenograft tumors in immunodeficient nu/nu mice^[4].
BXL0124 (0.03-0.1 μg/kg; i.p., p.o.; 6 days a week; 5 weeks) suppresses MCF10DCIS.com xenograft breast tumor growth by 66% (tumor weight reduction) via intraperitoneal administration at 0.1 μg/kg, while oral administration at 0.03 μg/kg and 0.1 μg/kg reduces tumor growth by 37% and 52% (tumor weight reduction), respectively, in SCID mice without hypercalcemic toxicity^[5].

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

Animal Model:

MMTV-ErbB2/neu transgenic (6-7 weeks old at study start)^[3]
Dosage:

0.3 μg/kg BW
Administration:

i.p.; three times weekly; 54 weeks
Result:

Reduced mean mammary tumor volume to 0.54 cm³, corresponding to 57% growth inhibition compared to controls.
Reduced mean tumor weight to 0.36 gram, corresponding to 54% inhibition compared to controls.
Reduced mean tumor multiplicity by 54% compared to controls.
Did not affect mouse body weight or induce hypercalcemic toxicity.
Down-regulated the phosphorylation of ErbB2, ERK, and AKT without altering total protein levels of these molecules in mammary tumors.
Reduced expression of the cell proliferation marker cyclin D1 in mammary tumors.

Animal Model:

nu/nu^[5]
Dosage:

0.1 μg/kg
Administration:

p.o.; 6 days a week; 5 weeks
Result:

Suppressed tumor size by 47% and tumor weight by 52%.
Markedly down-regulated protein expression of CD44 standard isoform (CD44s, 85 kDa), CD44 variant isoforms (CD44v, 100-250 kDa), CD44v3, and CD44v6 in xenograft tumors.
Reduced the proportion of cells with strong CD44 membrane staining (3+) from 48% to 15%, and reduced PCNA-positive cells from 82% to 62%.
No significant changes in body weight or serum calcium levels were observed.

Animal Model:

Severe combined immunodeficiency (SCID)^[5]
Dosage:

0.1 μg/kg (i.p.); 0.03 μg/kg (p.o.); 0.1 μg/kg (p.o.)
Administration:

i.p.; 6 days a week; 5 weeks; p.o.; 6 days a week; 5 weeks
Result:

Reduced average tumor volume by 75% and tumor weight by 66% at 0.1 μg/kg i.p..
Reduced tumor volume by 36% and tumor weight by 37% at 0.03 μg/kg p.o..
Reduced tumor volume by 49% and tumor weight by 52% at 0.1 μg/kg p.o..
No significant changes in body weight or serum calcium levels were observed with any dose or route tested.

Related Protocols :

Chemical Information

CAS No. 685141-23-5
Molecular Weight 606.68
Formula C₃₂H₄₄F₆O₄
SMILES

C[C@@]12[C@](CC[C@]2([H])[C@H](CCCC(C)(O)C)CC#CC(C(F)(F)F)(O)C(F)(F)F)([H])/C(CCC1)=C/C=C3C([C@H](C[C@@H](C\3)O)O)=C
Shipping

Room temperature in continental US; may vary elsewhere.
Storage

Please store the product under the recommended conditions in the Certificate of Analysis.

Purity & Documentation

Handling Instructions (2659 KB)

References

[1]. Shan NL, et al. Vitamin D compounds inhibit cancer stem-like cells and induce differentiation in triple negative breast cancer. The Journal of steroid biochemistry and molecular biology. 2017 Oct;173:122-129.

[2]. So JY, et al. HES1-mediated inhibition of Notch1 signaling by a Gemini vitamin D analog leads to decreased CD44(+)/CD24(-/low) tumor-initiating subpopulation in basal-like breast cancer. The Journal of steroid biochemistry and molecular biology. 2015 Apr;148:111-21.

[3]. Lee HJ, et al. Gemini vitamin D analog suppresses ErbB2-positive mammary tumor growth via inhibition of ErbB2/AKT/ERK signaling. The Journal of steroid biochemistry and molecular biology. 2010 Jul;121(1-2):408-12.

[4]. So JY, et al. Targeting CD44-STAT3 signaling by Gemini vitamin D analog leads to inhibition of invasion in basal-like breast cancer. PloS one. 2013;8(1):e54020.

[5]. So JY, et al. A novel Gemini vitamin D analog represses the expression of a stem cell marker CD44 in breast cancer. Molecular pharmacology. 2011 Mar;79(3):360-7.

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BXL0124

All NF-κB Isoforms

All STAT Isoforms

All JAK Isoforms