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Avagacestat (formerly known as BMS708163) is a potent, selective, orally bioactive γ-secretase inhibitor of Aβ40 and Aβ42 that may have anti-AD (Alzheimer's disease) properties. With IC50 values of 0.3 nM and 0.27 nM, respectively, it suppresses Aβ40 and Aβ42 and exhibits 193-fold selectivity against Notch. Currently under development as a treatment for mild to moderate AD and predementia, avagacestat is intended to selectively inhibit Aβ synthesis. It is possible that Avagacestat will result in fewer toxic adverse events than other less selective compounds because it produced up to 190-fold greater selectivity for Aβ synthesis than Notch processing in preclinical studies.
Physicochemical Properties
| Molecular Formula | C20H17CLF4N4O4S | |
| Molecular Weight | 520.88 | |
| Exact Mass | 520.059 | |
| Elemental Analysis | C, 46.12; H, 3.29; Cl, 6.81; F, 14.59; N, 10.76; O, 12.29; S, 6.16 | |
| CAS # | 1146699-66-2 | |
| Related CAS # |
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| PubChem CID | 46883536 | |
| Appearance | White to off-white solid powder | |
| Density | 1.5±0.1 g/cm3 | |
| Boiling Point | 652.3±65.0 °C at 760 mmHg | |
| Flash Point | 348.3±34.3 °C | |
| Vapour Pressure | 0.0±2.0 mmHg at 25°C | |
| Index of Refraction | 1.564 | |
| LogP | 4.89 | |
| Hydrogen Bond Donor Count | 1 | |
| Hydrogen Bond Acceptor Count | 11 | |
| Rotatable Bond Count | 9 | |
| Heavy Atom Count | 34 | |
| Complexity | 792 | |
| Defined Atom Stereocenter Count | 1 | |
| SMILES | 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 |
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| InChi Key | XEAOPVUAMONVLA-QGZVFWFLSA-N | |
| InChi Code | 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 | |
| Chemical Name | (2R)-2-[(4-chlorophenyl)sulfonyl-[[2-fluoro-4-(1,2,4-oxadiazol-3-yl)phenyl]methyl]amino]-5,5,5-trifluoropentanamide | |
| Synonyms | BMS-708163; BMS708163; Avagacestat; BMS 708163 | |
| HS Tariff Code | 2934.99.9001 | |
| Storage |
Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition | Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs) |
Biological Activity
| Targets |
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] |
| ln Vitro |
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]. 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] |
| ln Vivo |
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]. 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] |
| Enzyme Assay |
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] |
| Cell Assay |
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. 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] |
| Animal Protocol |
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 . 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] |
| ADME/Pharmacokinetics |
In male Sprague-Dawley rats, oral Avagacestat at 10 mg/kg showed an oral bioavailability of ~45%, a plasma elimination half-life (t₁/₂) of ~3.8 hours, a peak plasma concentration (Cmax) of 320 ng/mL (reached at 1.2 hours post-dose), and a volume of distribution (Vd) of ~2.0 L/kg [1] - In beagle dogs, oral Avagacestat at 0.5 mg/kg had a t₁/₂ of ~4.5 hours, a Cmax of 45 ng/mL (reached at 1.5 hours post-dose), and a brain-to-plasma concentration ratio of ~0.5 (measured 2 hours post-dose) [4] - In human AD patients (Phase II trial), oral Avagacestat (100 mg/day) reached a steady-state Cmax of ~180 ng/mL, with a t₁/₂ of ~5.2 hours [5] - Avagacestat has high plasma protein binding (>98%) in human, rat, and dog plasma (measured via ultrafiltration) [1] |
| Toxicity/Toxicokinetics |
In a 28-day rat toxicity study (oral Avagacestat at 5, 20, 60 mg/kg/day), the no-observed-adverse-effect level (NOAEL) was 20 mg/kg/day; at 60 mg/kg/day, mild skin hyperplasia and gastrointestinal mucosal inflammation were observed in 2/5 rats (reversible after treatment cessation) [1] - In beagle dogs (0.5 mg/kg oral for 14 days), no significant changes in serum ALT, AST, creatinine, or BUN levels were observed; no histopathological abnormalities in brain, liver, or kidney [4] - In the Phase II AD trial (n=252), the most common adverse events (AEs) with Avagacestat were diarrhea (18% [50 mg] vs. 25% [100 mg] vs. 8% placebo), fatigue (15% vs. 19% vs. 10%), and skin rash (12% vs. 18% vs. 5%); 5% of 100 mg/day patients discontinued due to AEs [5] |
| References |
[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. |
| Additional Infomation |
Avagacestat is an oxadiazole and a ring assembly. Avagacestat has been investigated for the basic science and treatment of Alzheimer Disease. Avagacestat (BMS-708163) is a small-molecule γ-secretase inhibitor initially developed for Alzheimer’s disease (AD) (via reducing Aβ production) and later investigated for EGFR inhibitor-resistant NSCLC (via reversing PI3K/Akt-mediated resistance) [1,2] - Unlike some γ-secretase inhibitors, Avagacestat directly targets the presenilin subunit of γ-secretase, ensuring potent inhibition of Aβ production; however, its lack of Notch-sparing activity increases the risk of Notch-related AEs (e.g., skin rash, gastrointestinal toxicity) [3,5] - Avagacestat completed Phase II clinical trials for mild-to-moderate AD but was discontinued due to lack of cognitive improvement and increased AEs at higher doses; it also showed preclinical efficacy in EGFR-resistant NSCLC but did not advance to late-stage clinical trials [2,5] - In canine models, Avagacestat is used as a tool to evaluate γ-secretase inhibitor efficacy in large animals, supporting translational research for AD therapeutics [4] |
Solubility Data
| Solubility (In Vitro) |
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| Solubility (In Vivo) |
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. (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 1.9198 mL | 9.5991 mL | 19.1983 mL | |
| 5 mM | 0.3840 mL | 1.9198 mL | 3.8397 mL | |
| 10 mM | 0.1920 mL | 0.9599 mL | 1.9198 mL |