FLAP antagonist | BI 665915

Target protein: 
FLAP
Probe Name: 
BI 665915
MOLECULAR WEIGHT [DA]: 
458.5
In stock: 
67485

Chemical structure

2D structure of BI 665915

Highlights

5‑Lipoxygenase Activating Protein (FLAP) is an important protein in the Leukotriene pathway. BI 665915 demonstrates excellent FLAP binding potency with an IC50 of below 10 nM and potent inhibition of LTB4 synthesis in human whole blood with an IC50 of below 100 nM. It is an excellent molecule for testing biological hypotheses in vitro and also in vivo.1 A favourable cross-species drug metabolism and DMPK profile and good selectivity make it an excellent molecule for testing biological hypotheses in vitro and in vivo. The compound is also active in mice which differentiates it from related compounds as highlighted in a recent review by D. Pettersen et al.2

With BI-0153 we also offer a structurally similar molecule which can be used as negative control for in vitro experiments due to significant weaker potency (670 nM).

3D structure of BI 665915

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Target information

5‑Lipoxygenase Activating Protein (FLAP) is an important protein in the Leukotriene (LT) pathway which acts as a partner of 5-lipoxygenase (5-LO) in the metabolism of arachidonic acid.3

Human FLAP in complex with leukotriene synthesis inhibitors (PDB code: 2q7r)

Human FLAP in complex with leukotriene synthesis inhibitors (PDB code: 2q7r)

Leukotriene (LT) Pathway

Leukotriene (LT) Pathway1

The membrane-attached 5-lipoxygenase activating protein (FLAP) binds to arachidonic acid (AA) and selectively transfers AA to 5-lipoxygenase (5-LO), which oxidizes AA to 5-hydroperoxyeicosatetraenoic acid (5-HpETE) followed by a dehydration to LTA4.1,3

Leukotrienes (LTs) are a family of eicosanoid proinflammatory mediators that are biosynthesized from arachidonic acid (AA) via oxidative metabolism.3

The leukotriene pathway constitutes a series of events underlying the inflammatory components of several diseases such as asthma, allergy, and atherosclerosis.3,5,6

More information about the target can be found in the following J. Med. Chem. publication1 by Hidenori Takahashi et al. and references cited therein.

In vitro Activity

BI 665915 shows a high potency (IC50 = 1.7 nM in the FLAP binding assay).

In vitro activity table

Probe name / Negative control

BI 665915

BI-0153

MW [Da]

459

430

FLAP binding (IC50) [nM]a

1.7

670

FLAP Functional inhibition in human whole blood (IC50) [nM]b

45

>5000

FLAP Functional inhibition in mouse whole blood (IC50) [nM]c

4800

n.d.

a Binding assay; geometric mean values (n ≥ 3), each determined from duplicate 10-point concentration−response curves;

b Human whole blood assays; geometric mean values (n ≥ 3), each determined from duplicate 10-point concentration−response curves;

c Mouse whole blood assays performed using the same protocol as that for the hWB assay; geometric mean values (n ≥ 3), each determined from duplicate 8-point concentration−response curves;

In vitro DMPK parameters

In vitro DMPK parameters table

Probe name / Negative control

BI 665915

BI-0153a

Solubility @ pH 6.8 [µg/ml]

48

>43

CACO permeability @ pH 7.4 [*10-6 cm/s]

34

n.d.

CACO efflux ratio

1.9

n.d.

Human hepatocyte clearance [%QH]

41

n.d.

Plasma protein binding human [%QH]

95.3

n.d.

a Please refer to the section negative control

In vivo DMPK parameters

BI 665915 was evaluated in rats, dogs, and cynomolgus monkeys (see table). The compound showed low iv plasma clearance over the three species and a good bioavailability of 45 to 63%.
In mice high exposures were observed at a dose of 100 mg/kg (AUC0-inf = 436,000 nM*h).

In vivo DMPK parameters of BI 665915 in the rat, dog, and cynomolgus monkeya

BI 665915

rat

dog

monkey

CL [%QH]b,c

7.0

2.8

3.6

Mean residence time after iv dose (l/kg)b

3.1

23

4.8

F [%]b

63

58

45

Vss [l/kg]b

0.9

1.2

0.5

a Dose = iv, 1 mg/kg; dosing vehicle, 70% PEG; po, 10 mg/kg; dosing suspension vehicle, 0.5% methyl cellulose/0.015% Tween; all DMPK parameters were determined after 11-time point blood sampling (0, 5, 15, 30 min, 1, 2, 4, 6, 8, 12, and 24 h) per iv or po dose.

b Mean values (n = 3).

c Value represents the percentage of hepatic blood flow.

In vivo pharmacology

BI 665915 shows an attractive DMPK profile and therefore was tested in a mouse ex vivo model of mechanism engagement. Blood samples were stimulated with calcimycin, and the levels of LTB4 were measured. BI 665915 demonstrated dose-dependent LTB4 production inhibition in mouse whole blood, 2 h after a single oral dose.1

Negative control

MOLECULAR WEIGHT OF NEGATIVE CONTROL [DA]: 
430.5

Negative control

Negative control

The closely related analogue BI-0153 can be used as an in vitro negative control

Selectivity

Extensive external screens covering 751 targets did not give strong hits (see supplementary data section)

Invitrogen® panel: 546 kinases < 30% inhibition @ 3µM
Panlabs® External screen covering 68 targets @ 10 µM
Cerep® External screen covering 137 targets @ 20 µM

Selectivity data available

Probe name

BI 665915

Cerep®

Yes

Eurofins-Panlabs®

Yes

Invitrogen®

Yes

DiscoverX®

No

Dundee

No

Download selectivity data: 

reference molecules

For a recent review on FLAP inhibitors see Reference 2

Summary

BI 665915 demonstrates nanomolar FLAP binding potency and is a molecule suitable for testing biological hypotheses in vitro and also in vivo.

Supplementary data

References

  1. Synthesis, SAR, and Series Evolution of Novel Oxadiazole-Containing 5-Lipoxygenase Activating Protein Inhibitors: Discovery of 2-[4-(3-{(R)-1-[4-(2-Amino-pyrimidin-5-yl)-phenyl]-1-cyclopropyl-ethyl}-[1,2,4]oxadiazol-5-yl)-pyrazol-1-yl]-N,N-dimethyl-acetam

    Hidenori Takahashi et al.

    J. Med. Chem. 2015;58:1669-1690.

  2. Recent advances for FLAP inhibitors

    Daniel Pettersen, Öjvind Davidsson, Carl Whatling

    Bioorg. Med. Chem. Lett. 2015;25:2607-2612.

  3. Funk Prostaglandins and Leukotrienes: Advances in Eicosanoid Biology

    Colin D.

    Science 2001;294:1791-1875.

  4. Crystal Structure of Inhibitor-Bound Human 5-Lipoxygenase–Activating Protein

    Andrew D. Ferguson, Brian M. McKeever, Shihua Xu, Douglas Wisniewski, Douglas K. Miller, Ting-Ting Yamin, Robert H. Spencer, Lin Chu, Feroze Ujjainwalla, Barry R. Cunningham, Jilly F. Evans, Joseph W. Becker

    Science 2007;317:510-512.

  5. Treatment of Asthma with Drugs Modifying the Leukotriene Pathway

    Jeffery M. Drazen, Elliot Israel, Paul M. O'Byrne

    Science 2007;317:510-512.

  6. The 5 lipoxygenase system in the vasculature: Emerging role in health and disease

    Etty Osher, Gary Weisinger, Rona Limor, Karen Tordjman, Naftali Stern

    Mol. Cell. Endcrinol. 2006;252:201−206.

  7. WO2013113799 A1

    Lars Anders Bylock