Notch1 (IC50 = 1.6 nM); Notch2 (IC50 = 0.7 nM); Notch3 (IC50 = 3.4 nM); Notch4 (IC50 = 3.4 nM)
BMS-906024 is a potent and selective inhibitor of the gamma secretase complex, which prevents the proteolytic activation of all four Notch receptors (Notch1, Notch2, Notch3, Notch4). The compound demonstrates a low nanomolar half-maximal inhibitory concentration (IC50) in in vitro enzyme assays and cellular Notch reporter assays.[1]

All six lung cancer cell lines have lower Notch1 ICD levels when exposed to BMS-906024 (5-100 nM; 72 hours). Hes1 cryopreservation was reported and total Notch1 was unaffected by BMS-906024 at 100 nM [1]. BMS-906024 inhibited triple negative medium (MDA-MB-468) and blank (TALL-1) cells in the experiment at an IC50 of around 4 nM [2]. BMS-906024 (100 nM; for 72 hours) improves paclitaxel Western Blot analysis [1].

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BMS-906024 potently inhibits Notch signaling in NSCLC cell lines. Treatment with concentrations as low as 5 nM reduced levels of the activated Notch1 intracellular domain (N1ICD), with maximal depletion observed at 50-100 nM. At 100 nM, it had no effect on total Notch1 protein but downregulated the mRNA expression of Hes1, a canonical Notch target gene, confirming functional inhibition of the pathway.[1]
BMS-906024 alone was not cytotoxic at concentrations up to 800 nM across all 31 NSCLC cell lines tested. However, it significantly enhanced the cytotoxicity of paclitaxel in vitro. The synergy was quantified using the Combination Index (CI) method (Chou-Talalay). The mean CI value for the combination of BMS-906024 (100 nM) and paclitaxel across 31 NSCLC cell lines was significantly lower (indicating greater synergy) than that with cisplatin (mean CI = 0.54 vs. 0.85, P = 0.01 for initial 14 lines). Synergy (CI < 0.7) was particularly strong and consistent in KRAS- and BRAF-wildtype (WT) cell lines (mean CI = 0.43) compared to KRAS- or BRAF-mutant cell lines (mean CI = 0.90, P = 0.003). The IC50 of paclitaxel significantly decreased when combined with BMS-906024 in the KRAS/BRAF-WT subgroup.[1]
Transient overexpression of activated Notch1 ICD or Notch3 ICD did not abrogate the synergy between BMS-906024 and paclitaxel, suggesting that sensitization may not be mediated solely through inhibition of these individual receptors.[1]
Knockdown of mutant KRAS (G12C) using siRNA in heterozygous mutant cell lines increased the synergy between BMS-906024 and paclitaxel.[1]

BMS-906024 (8.5 mg/kg; lateral gavage; days 1 to 4 per week for 3 weeks) significantly increased the tumor growth inhibitory effects of paclitaxel (36 mg/kg). BMS-906024 (?IV/PO) has T1/2 of 4.6/5.3 hours, Cmax of 1/0.3 μM, AUC. BMS-906024 enhances paclitaxel-mediated NSCLC cytotoxicity in vivo through a combination of inhibiting proliferation and promoting cell sealing in a p21- and p57-independent manner [1].

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In vivo studies using NSCLC cell line-derived xenografts and a patient-derived xenograft (PDX) model demonstrated that BMS-906024 significantly enhanced the antitumor activity of paclitaxel. In KRAS/BRAF-WT models (PDX-T42 and H838 xenografts), the combination of BMS-906024 and paclitaxel resulted in significantly greater tumor growth inhibition compared to either agent alone. Kaplan-Meier analysis showed significantly longer progression-free survival in the combination group. Similar enhancement was observed in some KRAS-mutant xenograft models (SKLU1, HCC44) in a manner consistent with their in vitro synergy profiles.[1]
The enhanced antitumor effect was associated with decreased cell proliferation (reduced Ki67 positivity) and increased apoptosis (elevated TUNEL positivity) in tumor tissues from combination-treated mice compared to paclitaxel alone.[1]

Human constructs were generated by PCR using standard molecular biology techniques and verified by sequencing. All constructs were generated in the pCDNA3.1+ Hyg vector (Invitrogen, Carlsbad, CA) and contain an N-terminal signal sequence, Notch coding sequence and a C-terminal c-myc tag (7 copies of EQKLISEEDL). The Notch coding sequence included begins N-terminal to the putative S2 cleavage site and contains the transmembrane and cytoplasmic domains. Coding sequence in the human Notch1 construct includes amino acids 1714-2555 (Accession NP_060087.3); a M1737V mutation within the transmembrane domain was added to suppress internal translation initiation. Coding sequence in the human Notch-2 construct includes amino acids 1645-2471 (Accession NP_077719.2). Coding sequence in the human Notch-3 construct includes amino acids 1622-2321 (Accession NP_000426.2). Coding sequence in the human Notch-4 construct includes amino acids 1415-2003 (Accession NP_004548.3).[2]

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Notch-CBF1 transactivation assay: HeLa cells were transiently cotransfected with plasmids containing truncated Notch1-4 receptors and a luciferase reporter vector with CBF1 binding sites. Cells were tested for Notch-CBF1 activity in the presence or absence of test compounds. IC50 values were determined from three independent experiments. [2]

Western Blot analysis [1]
Cell Types: NSCLC cell lines (A549, H358, H1975, H2444, H1792, HCC44)
Tested Concentrations: 5, 10, 25, 50, 100 nM
Incubation Duration: 72 hrs (hours).
Experimental Results: Notch1 ICD levels were diminished in all six lung cancer cell lines tested at concentrations as low as 5 nM, with maximum depletion concentrations of 50-100 nM.

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To assess chemosensitivity, cells were plated in 96-well plates and treated for 72 hours with a range of concentrations of cisplatin or paclitaxel (0.25x to 8x IC50), with or without a fixed 100 nM concentration of BMS-906024. Cell viability was measured using the CellTiter 96 Aqueous Non-Radioactive Cell Proliferation MTS Assay. Dose-response curves were generated, and IC50 values were calculated. Synergy was assessed using the Chou-Talalay method to calculate the Combination Index (CI) with CalcuSyn software.[1]
For Notch inhibition validation, cells were treated with varying concentrations of BMS-906024 (e.g., 5-100 nM) for 72 hours, followed by Western blot analysis to detect levels of Notch1 ICD and total Notch1. Downregulation of Hes1 mRNA was assessed by qRT-PCR after 24 hours of treatment with BMS-906024.[1]
For KRAS knockdown experiments, cells were transfected with 60 nM of a KRAS-G12C specific siRNA or non-targeting control siRNA using a lipid-based transfection reagent. Twenty-four hours post-transfection, cells were plated for MTS assays to evaluate changes in synergy.[1]
For exogenous Notch overexpression, cells were transfected with plasmids encoding Notch1 ICD or Notch3 ICD (or empty vector controls) using a transfection reagent. Twenty-four hours later, cells were plated for MTS assays to test if overexpression altered the synergy between BMS-906024 and paclitaxel. Parallel samples were analyzed by immunoblotting to confirm overexpression.[1]

Formulation: BMS-906024 was formulated in 10% vitamin E TPGS, 10% ethanol, and 80% PEG300 for in vivo studies.[1]

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Animal/Disease Models: 6 to 12 weeks old female NOD scid gamma (NSG) mice with KRAS- and BRAF -WT PDX-T42 xenograft[1]
Doses: 8.5 mg/kg
Route of Administration: 3.4/1.9 μM[2] hrs (hrs (hours)). po (oral gavage); days 1 to 4 per week for 3 weeks.
Experimental Results: Dramatically enhanced the tumor growth inhibitory effect of paclitaxel (36 mg/kg), but had no significant effect on cisplatin (2 mg/kg) treatment.

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Animal/Disease Models: Mouse[2]
Doses: 1 mg/kg (pharmacokinetic/PK/PK analysis)
Route of Administration: intravenous (iv) (iv)injection or oral administration
Experimental Results: T1/2 is 4.6/5.3 hrs (hrs (hours)), Cmax is 1/0.3 μM, AUC is 3.4 /1.9 μM • IV/PO hour.

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Female NOD scid gamma (NSG) mice, 6-12 weeks old, were used.
For efficacy studies, tumor xenografts were established by subcutaneous injection of NSCLC cell lines (e.g., HCC44, SKLU1, H838, A549) mixed with Matrigel, or by surgical implantation of PDX fragments (T-042). When the average tumor volume reached approximately 100 mm³, mice were randomized into treatment groups (6-8 mice per group).
BMS-906024 was formulated in 10% vitamin E TPGS, 10% ethanol, and 80% PEG300. It was administered by oral gavage (p.o.) at a dose of 8.5 mg/kg, on days 1 through 4 of each week for 3 weeks.
Paclitaxel was diluted in 0.9% sodium chloride and administered by intraperitoneal injection (i.p.) on day 1 of each week for 3 weeks. The specific paclitaxel dose varied per model and was determined in preliminary studies as the dose that reduced tumor growth by approximately 50% after 3 weeks.
In combination groups, BMS-906024 was given at least one hour after paclitaxel on days when both drugs were administered to avoid potential drug-drug interactions.
Tumor volume was measured twice weekly. Mice were euthanized when tumor size reached predefined endpoints or at the end of the study for tissue harvest.[1]
A separate tolerability study in non-tumor-bearing NSG mice evaluated BMS-906024 doses of 3.5, 4.5, 5.5, 6.5, and 7.5 mg/kg (p.o., days 1-4 weekly for 3 weeks). No dose-limiting toxicity was observed, supporting the use of the 8.5 mg/kg dose in efficacy studies.[1]

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Female NOD scid gamma (NSG) mice, 6-12 weeks old, were used.
For efficacy studies, tumor xenografts were established by subcutaneous injection of NSCLC cell lines (e.g., HCC44, SKLU1, H838, A549) mixed with Matrigel, or by surgical implantation of PDX fragments (T-042). When the average tumor volume reached approximately 100 mm³, mice were randomized into treatment groups (6-8 mice per group).
BMS-906024 was formulated in 10% vitamin E TPGS, 10% ethanol, and 80% PEG300. It was administered by oral gavage (p.o.) at a dose of 8.5 mg/kg, on days 1 through 4 of each week for 3 weeks.
Paclitaxel was diluted in 0.9% sodium chloride and administered by intraperitoneal injection (i.p.) on day 1 of each week for 3 weeks. The specific paclitaxel dose varied per model and was determined in preliminary studies as the dose that reduced tumor growth by approximately 50% after 3 weeks.
In combination groups, BMS-906024 was given at least one hour after paclitaxel on days when both drugs were administered to avoid potential drug-drug interactions.
Tumor volume was measured twice weekly. Mice were euthanized when tumor size reached predefined endpoints or at the end of the study for tissue harvest.[1]
A separate tolerability study in non-tumor-bearing NSG mice evaluated BMS-906024 doses of 3.5, 4.5, 5.5, 6.5, and 7.5 mg/kg (p.o., days 1-4 weekly for 3 weeks). No dose-limiting toxicity was observed, supporting the use of the 8.5 mg/kg dose in efficacy studies.[1]

BMS-906024 showed good intrinsic permeability in Caco-2 cells (104 nm/s) and was well absorbed orally.
Plasma half-life (t1/2) ranged from 4.6 h (mouse) to 51.1 h (dog) after intravenous administration.
Oral bioavailability (Fpo) ranged from 29% in dogs to 57% in mice.
Systemic clearance was < 10% of hepatic blood flow; volume of distribution (Vss) exceeded total body water, indicating extensive extravascular distribution.
Plasma protein binding was 88.7% in human serum and 95.9%, 94.3%, 95.4%, and 87.6% in mouse, rat, dog, and cynomolgus monkey serum, respectively. [2]

In the cited tolerability study in mice, doses of BMS-906024 up to 7.5 mg/kg (p.o., days 1-4 weekly for 3 weeks) showed no dose-limiting toxicity. In the efficacy studies, the combination of BMS-906024 (8.5 mg/kg) with paclitaxel was well-tolerated, causing no significant weight loss, diarrhea, or signs of distress.[1]

[1]. Gamma Secretase Inhibition by BMS-906024 Enhances Efficacy of Paclitaxel in Lung Adenocarcinoma. Mol Cancer Ther. 2017 Dec;16(12):2759-2769.

[2]. Discovery of Clinical Candidate BMS-906024: A Potent Pan-Notch Inhibitor for the Treatment of Leukemia and Solid Tumors. ACS Med Chem Lett. 2015 Mar 11;6(5):523-7.

(2S,3R)-N'-[(3S)-1-methyl-2-oxo-5-phenyl-3H-1,4-benzodiazepine-3-yl]-2,3-bis(3,3,3-trifluoropropyl)succinylamine is a primary, secondary, and tertiary carboxamide. Osulgastat (BMS-906024) has been used in clinical trials for the treatment of cancer, lymphoblastic leukemia, acute T-cell lymphoma, and precursor T-cell lymphoblastic lymphoma. Osulgastat is a small molecule γ-secretase (GS) and pan-Notch inhibitor with potential antitumor activity. After intravenous injection, osulgastat binds to GS and blocks Notch receptor activation, which may inhibit the proliferation of tumor cells with overactive Notch pathways. The integrated membrane protein GS is a multi-subunit protease complex that cleaves specific residues within the transmembrane domain of single-transmembrane proteins (such as the Notch receptor), thereby activating these proteins. Overactivation of the Notch signaling pathway (usually caused by activating mutations) is associated with increased cell proliferation and poor prognosis in certain tumor types.

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BMS-906024 is a clinical-stage gamma secretase inhibitor (GSI) originally developed from Alzheimer's disease research but repurposed for oncology.
It is described as a potent pan-Notch inhibitor that prevents the activation of all four Notch receptors.
The study identifies KRAS and BRAF wildtype status as a potential predictive biomarker for enhanced synergy between BMS-906024 and paclitaxel in lung adenocarcinoma. Within KRAS/BRAF-mutant tumors, TP53 mutation/null status and a low H2O2 pathway activity signature were also correlated with synergy.
The proposed mechanism for synergy involves BMS-906024 inhibiting Notch signaling, which may downregulate EGFR expression and subsequently impact the KRAS/MAPK pathway, particularly in KRAS/BRAF-WT contexts. Additionally, paclitaxel was found to decrease KRAS protein levels, and knockdown of mutant KRAS enhanced synergy.
BMS-906024 was under evaluation in Phase 1 clinical trials for leukemia and solid tumors at the time of this publication.[1]

C26H26F6N4O3

556.500067234039

556.19

C, 56.11; H, 4.71; F, 20.48; N, 10.07; O, 8.63

1401066-79-2

66550890

Typically exists as White to off-white solids at room temperature

1.4±0.1 g/cm3

726.3±60.0 °C at 760 mmHg

393.0±32.9 °C

0.0±2.4 mmHg at 25°C

1.558

2.1

2

10

9

39

916

3

CN1C2=CC=CC=C2C(C3=CC=CC=C3)=N[C@H](NC([C@@H]([C@@H](C(N)=O)CCC(F)(F)F)CCC(F)(F)F)=O)C1=O

AYOUDDAETNMCBW-GSHUGGBRSA-N

InChI=1S/C26H26F6N4O3/c1-36-19-10-6-5-9-18(19)20(15-7-3-2-4-8-15)34-22(24(36)39)35-23(38)17(12-14-26(30,31)32)16(21(33)37)11-13-25(27,28)29/h2-10,16-17,22H,11-14H2,1H3,(H2,33,37)(H,35,38)/t16-,17+,22+/m0/s1

(2R,3S)-N1-((S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)-2,3-bis(3,3,3-trifluoropropyl)succinamide

BMS906024; BMS 906024; Osugacestat; BMS 906024; Osugacestat [USAN]; AL101; (2R,3S)-N1-((S)-1-methyl-2-oxo-5-phenyl-2,3-dihydro-1H-benzo[e][1,4]diazepin-3-yl)-2,3-bis(3,3,3-trifluoropropyl)succinamide; DRL23N424R; BMS-906024.

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~9.5 mg/mL (~17.07 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)