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Identification and Preclinical Pharmacology of the γ-Secretase Modulator BMS-869780.

Toyn JH, Thompson LA, Lentz KA, Meredith JE, Burton CR, Sankaranararyanan S, Guss V, Hall T, Iben LG, Krause CM, Krause R, Lin XA, Pierdomenico M, Polson C, Robertson AS, Denton RR, Grace JE, Morrison J, Raybon J, Zhuo X, Snow K, Padmanabha R, Agler M, Esposito K, Harden D, Prack M, Varma S, Wong V, Zhu Y, Zvyaga T, Gerritz S, Marcin LR, Higgins MA, Shi J, Wei C, Cantone JL, Drexler DM, Macor JE, Olson RE, Ahlijanian MK, Albright CF - Int J Alzheimers Dis (2014)

Bottom Line: Off-target and safety margins were then based on comparisons to the predicted exposure required for robust Aβ1-42 lowering.Because of insufficient safety predictions and the relatively high predicted human daily dose of 700 mg, further evaluation of BMS-869780 as a potential clinical candidate was discontinued.Nevertheless, BMS-869780 demonstrates the potential of the GSM approach for robust lowering of brain Aβ1-42 without Notch-related side effects.

View Article: PubMed Central - PubMed

Affiliation: Exploratory Biology and Genomics, Bristol-Myers Squibb Research and Development, 5 Research Parkway, Wallingford, CT 06492, USA.

ABSTRACT
Alzheimer's disease is the most prevalent cause of dementia and is associated with accumulation of amyloid-β peptide (Aβ), particularly the 42-amino acid Aβ1-42, in the brain. Aβ1-42 levels can be decreased by γ-secretase modulators (GSM), which are small molecules that modulate γ-secretase, an enzyme essential for Aβ production. BMS-869780 is a potent GSM that decreased Aβ1-42 and Aβ1-40 and increased Aβ1-37 and Aβ1-38, without inhibiting overall levels of Aβ peptides or other APP processing intermediates. BMS-869780 also did not inhibit Notch processing by γ-secretase and lowered brain Aβ1-42 without evidence of Notch-related side effects in rats. Human pharmacokinetic (PK) parameters were predicted through allometric scaling of PK in rat, dog, and monkey and were combined with the rat pharmacodynamic (PD) parameters to predict the relationship between BMS-869780 dose, exposure and Aβ1-42 levels in human. Off-target and safety margins were then based on comparisons to the predicted exposure required for robust Aβ1-42 lowering. Because of insufficient safety predictions and the relatively high predicted human daily dose of 700 mg, further evaluation of BMS-869780 as a potential clinical candidate was discontinued. Nevertheless, BMS-869780 demonstrates the potential of the GSM approach for robust lowering of brain Aβ1-42 without Notch-related side effects.

No MeSH data available.


Related in: MedlinePlus

BMS-869780 did not inhibit Notch cleavage in vitro. (a) HeLa cell cultures were transfected with mNotchΔE and CBF1-luciferase reporter constructs, treated overnight with BMS-869780 (●), BMS-299897 (▲), or BMS-433796 (■), and luciferase assays were carried out. (b) HeLa cell cultures were transfected with mNotchΔ1865, treated with compounds overnight, and cell extracts were evaluated by western blot using anti-c-myc-HRP conjugate. Lanes 1 and 9: DMSO (0.1%) vehicle. Lane 2: BMS-299897 at 1 μM. Lane 3: BMS-433796 at 0.3 μM. Lanes 4–8: BMS-869780 at 0.1, 0.3, 1, 3, and 10 μM, respectively.
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fig6: BMS-869780 did not inhibit Notch cleavage in vitro. (a) HeLa cell cultures were transfected with mNotchΔE and CBF1-luciferase reporter constructs, treated overnight with BMS-869780 (●), BMS-299897 (▲), or BMS-433796 (■), and luciferase assays were carried out. (b) HeLa cell cultures were transfected with mNotchΔ1865, treated with compounds overnight, and cell extracts were evaluated by western blot using anti-c-myc-HRP conjugate. Lanes 1 and 9: DMSO (0.1%) vehicle. Lane 2: BMS-299897 at 1 μM. Lane 3: BMS-433796 at 0.3 μM. Lanes 4–8: BMS-869780 at 0.1, 0.3, 1, 3, and 10 μM, respectively.

Mentions: The effect of BMS-869780 on Notch processing was evaluated using transcriptional reporter assays and western blotting of NICD levels. HeLa cell cultures were transfected with mNotch1ΔE and CBF1-luciferase reporter constructs and treated with BMS-869780. In most replicates of this experiment, inhibition of luciferase reporter occurred with IC50 > 10 μM. In contrast, the GSIs BMS-299897 and BMS-433796 robustly inhibited luciferase activation, with IC50 = 340 nM and 2.1 nM, respectively (Figure 6(a)). The IC50 values for Notch-dependent luciferase expression are summarized in Table 1. To further evaluate the effect of BMS-869780 on Notch processing, western blots of cell cultures were carried out using mNotch1Δ1865, a truncated version of mNotch1ΔE that facilitates separation of NICD product from mNotch1 substrate on western blots [68]. HeLa cell cultures were transfected with mNotch1Δ1865 and treated overnight with compounds before western blotting. BMS-869780 had no effect on NICD levels at concentrations up to 3 μM, whereas the GSIs, BMS-299897, and BMS-433796, greatly reduced NICD (Figure 6(b)). Furthermore, the GSIs caused an increased level of mNotch1Δ1865 substrate, most likely due to inhibition of its turnover by γ-secretase, as previously noted by Blat et al. [68]. In contrast, 10 μM BMS-869780 did not increase mNotch1Δ1865 levels (Figure 6(b)), suggesting that the concomitant loss of NICD at 10 μM was due to a nonspecific effect, rather than inhibition of mNotch1Δ1865 turnover. This was also consistent with the observation of detached and dead cells in the presence of 10 μM BMS-869780.


Identification and Preclinical Pharmacology of the γ-Secretase Modulator BMS-869780.

Toyn JH, Thompson LA, Lentz KA, Meredith JE, Burton CR, Sankaranararyanan S, Guss V, Hall T, Iben LG, Krause CM, Krause R, Lin XA, Pierdomenico M, Polson C, Robertson AS, Denton RR, Grace JE, Morrison J, Raybon J, Zhuo X, Snow K, Padmanabha R, Agler M, Esposito K, Harden D, Prack M, Varma S, Wong V, Zhu Y, Zvyaga T, Gerritz S, Marcin LR, Higgins MA, Shi J, Wei C, Cantone JL, Drexler DM, Macor JE, Olson RE, Ahlijanian MK, Albright CF - Int J Alzheimers Dis (2014)

BMS-869780 did not inhibit Notch cleavage in vitro. (a) HeLa cell cultures were transfected with mNotchΔE and CBF1-luciferase reporter constructs, treated overnight with BMS-869780 (●), BMS-299897 (▲), or BMS-433796 (■), and luciferase assays were carried out. (b) HeLa cell cultures were transfected with mNotchΔ1865, treated with compounds overnight, and cell extracts were evaluated by western blot using anti-c-myc-HRP conjugate. Lanes 1 and 9: DMSO (0.1%) vehicle. Lane 2: BMS-299897 at 1 μM. Lane 3: BMS-433796 at 0.3 μM. Lanes 4–8: BMS-869780 at 0.1, 0.3, 1, 3, and 10 μM, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4109680&req=5

fig6: BMS-869780 did not inhibit Notch cleavage in vitro. (a) HeLa cell cultures were transfected with mNotchΔE and CBF1-luciferase reporter constructs, treated overnight with BMS-869780 (●), BMS-299897 (▲), or BMS-433796 (■), and luciferase assays were carried out. (b) HeLa cell cultures were transfected with mNotchΔ1865, treated with compounds overnight, and cell extracts were evaluated by western blot using anti-c-myc-HRP conjugate. Lanes 1 and 9: DMSO (0.1%) vehicle. Lane 2: BMS-299897 at 1 μM. Lane 3: BMS-433796 at 0.3 μM. Lanes 4–8: BMS-869780 at 0.1, 0.3, 1, 3, and 10 μM, respectively.
Mentions: The effect of BMS-869780 on Notch processing was evaluated using transcriptional reporter assays and western blotting of NICD levels. HeLa cell cultures were transfected with mNotch1ΔE and CBF1-luciferase reporter constructs and treated with BMS-869780. In most replicates of this experiment, inhibition of luciferase reporter occurred with IC50 > 10 μM. In contrast, the GSIs BMS-299897 and BMS-433796 robustly inhibited luciferase activation, with IC50 = 340 nM and 2.1 nM, respectively (Figure 6(a)). The IC50 values for Notch-dependent luciferase expression are summarized in Table 1. To further evaluate the effect of BMS-869780 on Notch processing, western blots of cell cultures were carried out using mNotch1Δ1865, a truncated version of mNotch1ΔE that facilitates separation of NICD product from mNotch1 substrate on western blots [68]. HeLa cell cultures were transfected with mNotch1Δ1865 and treated overnight with compounds before western blotting. BMS-869780 had no effect on NICD levels at concentrations up to 3 μM, whereas the GSIs, BMS-299897, and BMS-433796, greatly reduced NICD (Figure 6(b)). Furthermore, the GSIs caused an increased level of mNotch1Δ1865 substrate, most likely due to inhibition of its turnover by γ-secretase, as previously noted by Blat et al. [68]. In contrast, 10 μM BMS-869780 did not increase mNotch1Δ1865 levels (Figure 6(b)), suggesting that the concomitant loss of NICD at 10 μM was due to a nonspecific effect, rather than inhibition of mNotch1Δ1865 turnover. This was also consistent with the observation of detached and dead cells in the presence of 10 μM BMS-869780.

Bottom Line: Off-target and safety margins were then based on comparisons to the predicted exposure required for robust Aβ1-42 lowering.Because of insufficient safety predictions and the relatively high predicted human daily dose of 700 mg, further evaluation of BMS-869780 as a potential clinical candidate was discontinued.Nevertheless, BMS-869780 demonstrates the potential of the GSM approach for robust lowering of brain Aβ1-42 without Notch-related side effects.

View Article: PubMed Central - PubMed

Affiliation: Exploratory Biology and Genomics, Bristol-Myers Squibb Research and Development, 5 Research Parkway, Wallingford, CT 06492, USA.

ABSTRACT
Alzheimer's disease is the most prevalent cause of dementia and is associated with accumulation of amyloid-β peptide (Aβ), particularly the 42-amino acid Aβ1-42, in the brain. Aβ1-42 levels can be decreased by γ-secretase modulators (GSM), which are small molecules that modulate γ-secretase, an enzyme essential for Aβ production. BMS-869780 is a potent GSM that decreased Aβ1-42 and Aβ1-40 and increased Aβ1-37 and Aβ1-38, without inhibiting overall levels of Aβ peptides or other APP processing intermediates. BMS-869780 also did not inhibit Notch processing by γ-secretase and lowered brain Aβ1-42 without evidence of Notch-related side effects in rats. Human pharmacokinetic (PK) parameters were predicted through allometric scaling of PK in rat, dog, and monkey and were combined with the rat pharmacodynamic (PD) parameters to predict the relationship between BMS-869780 dose, exposure and Aβ1-42 levels in human. Off-target and safety margins were then based on comparisons to the predicted exposure required for robust Aβ1-42 lowering. Because of insufficient safety predictions and the relatively high predicted human daily dose of 700 mg, further evaluation of BMS-869780 as a potential clinical candidate was discontinued. Nevertheless, BMS-869780 demonstrates the potential of the GSM approach for robust lowering of brain Aβ1-42 without Notch-related side effects.

No MeSH data available.


Related in: MedlinePlus