FAS inhibitor | BI 99179
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Highlights
The non-covalent inhibitor of type I fatty acid synthase (FAS) is a potent and selective compound suitable for in vitro and in vivo validation of FAS as a therapeutic target. The compound has significant peripheral and central exposure upon oral administration in rats.1 The optical antipode BI 99990 is offered as in vitro negative control.
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Target information
Mammalian type I fatty acid synthase (FAS) is a key enzyme for lipogenesis and highly expressed in lipogenic tissues. While most tissues, except liver and adipose tissue, have low levels of FAS expression and activity, FAS is over expressed in many cancers.2, 3
FAS inhibition could be a potential way to treat obesity.4
It has been reported that inhibitors of FAS reduce the production of sebum in sebocytes,5 suggesting topical FAS inhibition as a potential anti-acne approach.
The reported involvement of FAS in the mechanisms of viral infection and replication suggests that FAS inhibition could be applied as an antiviral principle8, 9, 10.
Structure of full length fatty acid synthase, (PDB code: 2vz9.pdb).6
Human Fatty Acid Synthase Psi/KR Tri-Domain with GSK2194069 bound next to NADPH (grey; PDB code: 4piv)
The two identical subunits each comprise an acyl carrier protein (ACP) domain and six different catalytic domains.
BI 99179 most probably binds to the ketoacyl reductase domain (evidence: inhouse enzymatic data and analogy to the published co-crystal structure of the human KR domain with GSK2194069)7
In vitro Activity
BI 99179 inhibits the FAS enzyme isolated form HeLa cells with an IC50 of 79 nM. The optical antipode which can be used as a negative control shows an activity of >3000 nM in this assay.
In vitro activity table
|
Probe name / Negative control |
BI 99179 |
BI 99990 |
|
Inhibition of FASN* (IC50) [nM] |
79 |
>3000 |
|
Inhibition of [14C]acetate incorporation§ mouse N-42 cells (IC50) [nM] |
570 |
>10000 |
|
Inhibition of [14C]acetate incorporation§ hum. H1975 cells (IC50) [nM] |
180 |
>10000 |
|
Cytotox. (LDH release from U937 cells) (IC50) [nM] |
>30000 |
n.d. |
Cell lines:
* Human FAS enzyme isolated from HeLa cells
§ Cells incubated with compound for 1h, 14C-acetate in Krebs-Ringer-buffer incubation for 4h, Methanol:CHCl3 1:1 extraction, measurement in ß-counter;
e HEK membrane prep
In vitro DMPK parameters
|
Probe name / Negative Control |
BI 99179 |
BI 99990 |
|
Aqueous solubility @ pH 7 [µg/ml] |
>39 |
>39 |
|
CACO permeability @ pH 7.4 [*10-6 cm/s] |
94 |
n.d. |
|
CACO efflux ratio |
0.9 |
n.d. |
|
Microsomal stability – rat, human [% QH] |
<27 |
33 |
|
Plasma protein binding rat [% QH] |
97.6 |
n.d. |
In vivo DMPK parameters
|
Probe name / Negative control |
BI 99179 |
BI 99990 |
|
t1/2 [h] |
3.0 |
n.d. |
|
tmax [h] |
0.5 |
0.5 |
|
Cmax [nM] |
2110 |
317 |
|
AUC0–inf [nMh] |
9350 |
2160 |
|
F [%] |
46 |
n.d. |
|
CL [ml/min/kg] |
8.2 |
n.d. |
|
Vss [l/kg] |
1.6 |
n.d. |
|
Cbrain,2h [nM] |
1300 |
n.d. |
|
CCSF,2h [nM] |
50 |
n.d. |
Pharmacokinetic parameters of in male Han/Wistar rats *fasted upon oral application of 4 mg/kg
In vivo pharmacology
BI 99179 showed acute efficacy in rats
- Increased hypothalamic [Malonyl-CoA] (10 or 100 mg/kg; 2h /24h post dose (p.d.))
![hypothalamic [Malonyl-CoA] 2h/24h p.d.](/sites/default/files/figure-5-hypothalamic.PNG)
Hypothalamic [Malonyl-CoA] 2h/24h p.d.; cbr,unbound
- Decreased cumulative food intake 100 mg/kg; 24h p.d.
Pharmacologic POC in Han Wistar rats; Food intake with refeeding after 24h fast
BI 99179 showed adverse effects beginning at day 4 (30 mg/kg), day 9 (3 mg/kg)
- Reddened and swollen mouth (not seen with 50 mg/kg BI 99990)
- Reddened and swollen eye lids (not seen with 50 mg/kg BI 99990)
- Salivation (less pronounced with BI 99990)
- hair loss (less pronounced with BI 99990)
BI 99990 showed additional adverse effects at 50 mg/kg, therefore the compound is not suitable for further studies in rats.
Negative control
BI 99990 which serves as a negative control
The optical antipode BI 99990 can be used as negative control (IC50 FAS = >3000 nM). We tested BI 99990 also in vivo to assess if the observed adverse effects for BI 99719 are rather compound or target specific with the result that the adverse effects have not been observed or have been less pronounced with BI 99990. (For more details see in vivo pharmacology section.) BI 99990 showed additional adverse effects at 50 mg/kg, therefore the compound is not suitable for further studies in rats and should be only used in vitro.
Selectivity
No closely related mammalian proteins as potential off-targets.
Selectivity on non-related targets: External screen covering 30 targets: <20% inhibition @ 10 µM for all targets. (Please see Supplementary data for detailed information)
High selectivity confirmed in >100 in-house screens @ BI (including >10 enzymes, >10 GPCRs, >10 kinases, and 5 ion channels).
|
BI 99179 |
Selectivity data available |
|
Cerep® |
No |
|
Panlabs® |
Yes |
|
Invitrogen® |
No |
|
DiscoverX® |
No |
|
Dundee |
No |
reference molecules
See references 3 and 11
Summary
Our FAS in vivo tool compound BI 99179 is characterised by high potency, good selectivity and significant peripheral and central exposure upon oral administration in rats.
Supplementary data
References
Discovery of BI 99179, a potent and selective inhibitor of type I fatty acid synthase with central exposure
Kley J. T., Mack J., Hamilton B., Scheuerer S., Redemann N.
Bioorg. Med. Chem. Lett. 2011, 21, 5924-5927.
Fatty acid synthase as a potential therapeutic target in cancer
Flavin R., Peluso S., Nguyen P. L., Loda M.
Future Oncol. 2010, 6, 551-562.
IV Fatty acid synthase inhibitors: new directions for oncology
Kridel S. J., Lowther W. T., Pemble C. W.
Expert Opin. Invest. Drugs 2007, 16, 1817-1829.
Reduced Food Intake and Body Weight in Mice Treated with Fatty Acid Synthase Inhibitors
Loftus T. M., Jaworsky D. E., Frehywot G. L., Townsend C. A., Ronnett G. V., Lane M. D., Kuhajda F. P.
Inhibitors Science 2000, 288, 2379.
U.S. Patent 53631
D’Arcangelis, A. D., et al.
2005
The Crystal Structure of a Mammalian Fatty Acid Synthase
Maier T., Leibundgut M., Ban N.
Science 2008, 321, 1315-1322.
A human fatty acid synthase inhibitor binds β-ketoacyl reductase in the keto-substrate site
Hardwicke M. A., Rendina A. R., Williams S. P., Moore M .L., Wang L., Krueger J. A., Plant R. N., Totoritis R. D., Zhang G., Briand J., Burkhart W. A., Brown K. K., Parrish C. A.
Nature Chemical Biology 2014, 10, 774-779.
Involvement of fatty acid synthase in dengue virus infection
Tongluan N., Nio Y., Akahori Y., Kim S., Watashi K., Wakita T., Hijikata M.
Virology Journal 2017, 14, 28
Direct Inhibition of Cellular Fatty Acid Synthase Impairs Replication of Respiratory Syncytial Virus and Other Respiratory Viruses
Ohol Y. M., Wang Z., Kemble G., Duke G.
PLoS One 2015, 10
Fatty Acid Synthase
Jones S. F., Infante J. R., Molecular Pathways
Clin. Cancer Res. 2015, 21, 5434-5438
Papers with our molecules. Published by you.
Mechanistic MALDI-TOF Cell-Based Assay for the Discovery of Potent and Specific Fatty Acid Synthase Inhibitors
Weigt D., Parrish CA., Krueger JA., Oleykowski CA., Rendina AR., Hopf C.
Cell Chamical Biology 2019, pii: S2451-9456(19)30205-3. open access
