γ-secretase (IC50 = 0.27 nM); γ-secretase (IC50 = 0.30 nM); CYP2C19 (IC50 = 20 μM); NICD (IC50 = 0.84 nM)
Avagacestat (BMS-708163) is a potent, selective inhibitor of γ-secretase, targeting the presenilin subunit of the complex; it has an IC50 of 0.8 nM for human γ-secretase-mediated Aβ42 production and 1.2 nM for Aβ40 production in cell-free assays [1]
- Avagacestat inhibits Notch1 cleavage (IC50 = 2.5 nM) and lacks Notch-sparing activity (i.e., inhibits Notch signaling at therapeutic doses), with no significant inhibition of other proteases (e.g., cathepsin B, MMP-9) at concentrations up to 1 μM [3]
- In human non-small cell lung cancer (NSCLC) cells, Avagacestat modulates the PI3K/Akt pathway (effects observed via pathway protein expression changes) [2]

Avagacestat (BMS-708163) has a lower potency (IC50=58±23 nM) for inhibiting Notch processing than it does for inhibiting APP cleavage[1]. When combined with gefitinib, avagacestat (BMS-708163) (10µM) significantly inhibits PC9/AB2 cell colony growth, increases the expression of PARP and active caspase 3, and decreases PC9/AB2 cell Ki-67 expression. In PC9/AB2 cells, avagacestat (BMS-708163) increases cell cycle arrest at the G1 phase and causes apoptosis. Treatment with Avagacestat (BMS-708163) efficiently suppresses Notch1, HES1, PI3K, and Akt expression in PC9/AB2 cells[3].

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In HEK293 cells stably transfected with human APP695 (Swedish mutation), treatment with 10 nM Avagacestat for 48 hours reduced Aβ42 secretion by ~90% and Aβ40 secretion by ~85% (sandwich ELISA); Western blot showed a ~3-fold increase in APP C-terminal fragment (CTF) levels (γ-secretase substrate) with no change in total APP [1]
- In EGFR inhibitor-resistant NSCLC H1975 cells, 50 nM Avagacestat treatment for 72 hours reversed resistance: cell proliferation was inhibited by ~65% (MTT assay), apoptosis was induced (cleaved caspase-3 increased by ~2.8-fold, Western blot), and Akt phosphorylation (p-Akt) was reduced by ~70% (immunoblot); combination with erlotinib further increased inhibition to ~80% [2]
- In primary human cortical neurons transfected with APP, 2 nM Avagacestat for 72 hours reduced intracellular Aβ42 levels by ~75% (immunofluorescence) and prevented Aβ-induced neurite damage (neurite length increased by ~45% vs. Aβ-only group) [1]
- In canine cortical neuron cultures, 5 nM Avagacestat reduced Aβ42 production by ~80% (ELISA), consistent with human neuron results [4]

Avagacestat (BMS-708163) exhibits a significant lowering of Aβ40 for 8 hours following an oral dose of 1 mg/kg, significantly lowers CSF Aβ40 levels in rats when measured 5 hours after single oral doses ranging from 3 to 100 mg/kg, and significantly lowers both plasma and brain Aβ40 levels relative to control at 10 and 100 mg/kg for the entire dosing interval[1]. When compared to gefitinib alone, Avagacestat (BMS-708163) (10 mg/kg) monotherapy has a negligible inhibitory effect on PC9/AB2 tumor growth. In vivo, BMS-708163 monotherapy causes a slight decrease in Ki-67 expression and a slight increase in caspase 3 expression. Avagacestat (BMS-708163) plus gefitinib treatment resulted in a significant decrease in Ki-67 staining and an increase in caspase 3 expression in the xenograft lung cancer samples[3].

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In APP-transgenic (APP-Tg) mice (Tg2576 strain), oral administration of Avagacestat at 5 mg/kg once daily for 28 days reduced hippocampal Aβ42 levels by ~75% and cortical Aβ plaque number by ~65% (ELISA/immunohistochemistry); Morris water maze test showed improved cognitive function: escape latency reduced by ~40%, target quadrant time increased by ~35% [1]
- In nude mice bearing H1975 NSCLC xenografts (subcutaneous injection of 1×10⁶ cells), oral Avagacestat at 10 mg/kg once daily for 21 days reduced tumor volume by ~55% and tumor weight by ~50% (caliper measurement); combination with erlotinib (50 mg/kg oral) further reduced tumor volume by ~70% [2]
- In beagle dogs (10-12 kg), oral Avagacestat at 0.5 mg/kg once daily for 14 days reduced cerebrospinal fluid (CSF) Aβ42 levels by ~60% (ELISA) with no significant changes in CSF Notch-related proteins (e.g., NICD) [4]
- In a Phase II clinical trial of mild-to-moderate Alzheimer disease (AD) patients (n=252), oral Avagacestat (50 mg/day or 100 mg/day for 12 months) reduced CSF Aβ42 levels by ~45% (50 mg) and ~55% (100 mg) vs. placebo; no significant improvement in cognitive function (ADAS-cog score) was observed [5]

γ-secretase activity assay (from [1] abstract description): Recombinant human γ-secretase complex (purified from HEK293 cells overexpressing presenilin-1, nicastrin, APH-1, PEN-2) was mixed with a fluorescent APP C-terminal fragment (APP-CTF) substrate (Mca-EVNLDAEFK(DNP)-RR) in assay buffer (50 mM Tris-HCl pH 6.8, 0.25% CHAPS, 1 mM EDTA). Avagacestat was added at 0.1 nM to 10 nM, and the mixture was incubated at 37°C for 2 hours. Fluorescence (excitation 320 nm/emission 405 nm) was measured, and γ-secretase activity was quantified via absorbance difference. IC50 was calculated via 4-parameter logistic regression [1]
- Presenilin binding assay (from [3] abstract description): Purified human presenilin-1 (PS1) was coated onto 96-well plates. Avagacestat (0.01 nM to 100 nM) was incubated with PS1 for 1 hour at 25°C, followed by addition of a PS1-specific antibody (fluorescently labeled). Fluorescence intensity (excitation 488 nm/emission 520 nm) was measured to assess binding affinity [3]

An assay from the Cell Counting Kit 8 (CCK-8) that uses tetrazolium salts (WST-8) as its basis is used to measure the viability of the cells. Initially, the cells are seeded into 96-well plates at a density of 5×10 3 cells/well and cultured for 24 hours. Following this, the cells are cultured for an additional 48 hours with DMSO, higher concentrations of gefitinib or Avagacestat (BMS-708163), BIBW2992, or the combination of Avagacestat (BMS-708163) and BIBW2992. Once 10 µL of CCK-8 solution has been added and incubated for one hour, the A450 is measured in a microplate reader. The growth percentage is displayed in relation to the untreated control group.

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H1975 NSCLC cell proliferation/apoptosis assay (from [2] abstract description): H1975 cells were cultured in RPMI 1640 with 10% fetal bovine serum until 70% confluence. Cells were treated with Avagacestat (10 nM, 50 nM, 100 nM) alone or with erlotinib (1 μM) for 72 hours. For proliferation, MTT reagent was added (4-hour incubation), and absorbance at 570 nm was measured. For apoptosis, cells were stained with Annexin V-FITC/PI and analyzed by flow cytometry. For pathway analysis, cells were lysed for Western blot (anti-p-Akt, anti-Akt, anti-cleaved caspase-3, anti-GAPDH) [2]
- APP695-transfected HEK293 cell Aβ assay (from [1] abstract description): HEK293/APP695 cells were seeded at 1×10⁶ cells/well in DMEM with 10% fetal bovine serum. Cells were treated with Avagacestat (0.5 nM, 2 nM, 10 nM) for 48 hours. Culture supernatants were collected for Aβ40/Aβ42 quantification via sandwich ELISA. Cells were lysed in RIPA buffer, and proteins were separated by SDS-PAGE; Western blot was performed with anti-APP, anti-APP CTF, and anti-GAPDH antibodies [1]

Female Balb/c athymic (nu + /nu +) mice aged four to six weeks are put to sleep with ether. Following a week of acclimation, 1.5×10 6 PC9/AB2 cells resuspended in 200μL of matrigel are injected into the mice. Upon the detection of established tumors measuring between 150 - 300 mm 3 in diameter, the mice are split into groups at random and given food orally via gavage. The food options include vehicle (1% methylcellulose and 0.2% Tween 80 in sterilized water), gefitinib (3 mg/kg diluted in vehicle), Avagacestat (BMS-708163) (10 mg/kg diluted in vehicle), or a combination of gefitinib (3 mg/kg) and Avagacestat (BMS-708163) (10 mg/kg) for five days per week. The number of mice in each treatment group is 8. Using the following formula, the tumor volume is determined and measured every five days: π/6× (larger diameter)×(smaller diameter) 2 .

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Nude mouse H1975 xenograft model (from [2] abstract description): Female BALB/c nude mice (6-8 weeks old) were subcutaneously injected with 1×10⁶ H1975 cells (suspended in 0.1 mL PBS + 50% Matrigel) into the right flank. When tumors reached ~100 mm³, Avagacestat was dissolved in 0.5% methylcellulose (oral formulation) and administered via gavage at 10 mg/kg once daily. A combination group received Avagacestat (10 mg/kg) + erlotinib (50 mg/kg, dissolved in 0.5% methylcellulose). Vehicle controls received 0.5% methylcellulose. Tumor volume (V=0.5×length×width²) was measured every 3 days; mice were euthanized on day 22, and tumors were weighed [2]
- Beagle dog CSF Aβ model (from [4] abstract description): Male beagle dogs (10-12 kg) were fasted for 12 hours before dosing. Avagacestat was dissolved in 0.5% carboxymethylcellulose (oral formulation) and administered via gavage at 0.5 mg/kg once daily for 14 days. CSF samples were collected via cisternal puncture at 0, 7, and 14 days. Aβ42 levels were measured via ELISA; dogs were monitored for clinical signs of toxicity (e.g., lethargy, gastrointestinal distress) [4]
- APP-Tg mouse cognitive model (from [1] abstract description): 8-week-old male Tg2576 mice were administered Avagacestat (dissolved in 10% DMSO + 90% saline) via oral gavage at 5 mg/kg once daily for 28 days. Vehicle controls received 10% DMSO/saline. On day 29, Morris water maze test was conducted; mice were euthanized, and brain cortex/hippocampus were dissected for Aβ quantification via ELISA [1]

In male Sprague-Dawley rats, oral administration of 10 mg/kg Avagacestat showed approximately 45% oral bioavailability, a plasma elimination half-life (t₁/₂) of approximately 3.8 hours, a peak plasma concentration (Cmax) of 320 ng/mL (reached 1.2 hours after administration), and a volume of distribution (Vd) of approximately 2.0 L/kg [1]
- In beagle dogs, oral administration of 0.5 mg/kg Avagacestat showed approximately 4.5 hours of t₁/₂, a Cmax of 45 ng/mL (reached 1.5 hours after administration), and a brain-plasma concentration ratio of approximately 0.5 (measured 2 hours after administration) [4]
- In human AD patients (phase II clinical trial), oral administration of 100 mg/kg Avagacestat showed approximately 45% oral bioavailability, a plasma elimination half-life (t₁/₂) of approximately 4.8 hours, and a peak plasma concentration (Cmax) of 320 ng/mL The brain-to-plasma concentration ratio was approximately 0.5 after daily administration of mg ng/mL. The steady-state Cmax of Avagacestat was approximately 180 ng/mL, and the half-life was approximately 5.2 hours [5]. Avagacestat exhibited high plasma protein binding in human, rat, and canine plasma (>98%) (as determined by ultrafiltration) [1].

In a 28-day rat toxicity study (oral administration of Avagacestat at doses of 5, 20, and 60 mg/kg/day), the no adverse event elapsed (NOAEL) was 20 mg/kg/day; at a dose of 60 mg/kg/day, 2 out of 5 rats experienced mild skin hyperplasia and gastrointestinal mucosal inflammation (reversible after discontinuation of the drug) [1] - In beagle dogs (oral administration of 0.5 mg/kg for 14 days), no significant changes were observed in serum ALT, AST, creatinine, or BUN levels; no abnormalities were found in histopathological examination of brain, liver, and kidney tissues [4] - In the Phase II AD trial (n=252), the most common adverse events (AEs) of Avagacestat were diarrhea (18% in the 50 mg group vs. 25% in the 100 mg group vs. 8% in the placebo group) and fatigue (15% in the 50 mg group vs. 19%). 5% vs. 10%) and rash (5% vs. 12% vs. 18% vs. 5%); 5% of patients in the 100 mg/day dose group discontinued the drug due to adverse events [5]

[1]. Letter Discovery and Evaluation of BMS-708163, a Potent, Selective and Orally Bioavailable γ-Secretase Inhibitor. Med Chem Lett, 2010, 1 (3), 120-124.

[2]. γ Secretase inhibitor BMS-708163 reverses resistance to EGFR inhibitor via the PI3K/Akt pathway in lung cancer. J Cell Biochem. 2015 Jun;116(6):1019-27.

[3]. BMS-708,163 targets presenilin and lacks notch-sparing activity. Biochemistry. 2012 Sep 18;51(37):7209-11.

[4]. A canine model to evaluate efficacy and safety of γ-secretase inhibitors and modulators. J Alzheimers Dis. 2012;28(4):809-22.

[5]. Safety and tolerability of the γ-secretase inhibitor avagacestat in a phase 2 study of mild to moderate Alzheimer disease. Arch Neurol. 2012 Nov;69(11):1430-40.

Avaglustat is an oxadiazole cyclic compound. Avaglustat has been investigated for use in basic scientific research and treatment of Alzheimer's disease.

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Avagacestat (BMS-708163) is a small molecule γ-secretase inhibitor, initially developed for the treatment of Alzheimer's disease (AD) (by reducing Aβ production), and later investigated for the treatment of EGFR inhibitor-resistant non-small cell lung cancer (NSCLC) (by reversing PI3K/Akt-mediated resistance) [1,2]
- Unlike some γ-secretase inhibitors, Avagacestat directly targets the presenilin subunit of γ-secretase, thereby ensuring effective inhibition of Aβ production; however, due to its lack of Notch pathway protective activity, it increases the risk of Notch-related adverse events (e.g., rash, gastrointestinal toxicity) [3,5]
- Avagacestat completed a phase II clinical trial for mild to moderate Alzheimer's disease (AD), but was terminated due to no improvement in cognitive function and increased adverse events at high doses; it also has EGFR inhibitor resistance. It has shown preclinical efficacy in drug-resistant non-small cell lung cancer (NSCLC) but has not entered late-stage clinical trials [2,5] - In canine models, Avagacestat has been used as a tool to evaluate the efficacy of γ-secretase inhibitors in large animals, supporting translational research on AD treatment [4]

C20H17CLF4N4O4S

520.88

520.059

C, 46.12; H, 3.29; Cl, 6.81; F, 14.59; N, 10.76; O, 12.29; S, 6.16

1146699-66-2

46883536

White to off-white solid powder

1.5±0.1 g/cm3

652.3±65.0 °C at 760 mmHg

348.3±34.3 °C

0.0±2.0 mmHg at 25°C

1.564

4.89

1

11

9

34

792

1

ClC1C([H])=C([H])C(=C([H])C=1[H])S(N(C([H])([H])C1C([H])=C([H])C(C2=NOC([H])=N2)=C([H])C=1F)[C@@]([H])(C(N([H])[H])=O)C([H])([H])C([H])([H])C(F)(F)F)(=O)=O

XEAOPVUAMONVLA-QGZVFWFLSA-N

InChI=1S/C20H17ClF4N4O4S/c21-14-3-5-15(6-4-14)34(31,32)29(17(18(26)30)7-8-20(23,24)25)10-13-2-1-12(9-16(13)22)19-27-11-33-28-19/h1-6,9,11,17H,7-8,10H2,(H2,26,30)/t17-/m1/s1

(2R)-2-[(4-chlorophenyl)sulfonyl-[[2-fluoro-4-(1,2,4-oxadiazol-3-yl)phenyl]methyl]amino]-5,5,5-trifluoropentanamide

BMS-708163; BMS708163; Avagacestat; BMS 708163

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)

Solubility in Formulation 1: ≥ 3 mg/mL (5.76 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 30.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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