The FASN inhibitor TVB-2640 is efficacious in a new 3D ...
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The FASN inhibitor TVB-2640 is efficacious in a new 3D human liver microtissue model of NASH TVB-2640 Fibrosis TVB - 2640 Steatosis FASN TVB -2640 TVB-2640 Steatosis Inflammation • Once-a-day oral small molecule • Potent with FASN EC 50 approx. 50 nM • Excellent and predictable human PK profile • Inhibits hepatic DNL up to 90% in human 13 C-acetate tracer study 1 • First-in-class FASN inhibitor Introduction • De novo lipogenesis (DNL) is upregulated in NASH and drives steatosis and lipotoxicity • Fatty acid synthase (FASN) is the last committed step in the DNL pathway and produces palmitate • Palmitate is not only a building block for lipids, but a signaling molecule that activates the inflammasome and TLR pathways, impacting inflammation and fibrosis in NASH Results in liver microtissue (LMT) model TVB-2640 inhibits three hallmarks of NASH; steatosis, inflammation and fibrosis Conclusions TVB-2640 reduced liver fat in NASH patients in Ph2 FASCINATE-1 trial • 61% patients treated with 50 mg TVB-2640 had ⩾ 30% reduction in liver fat by MRI-PDFF, vs 11% in placebo • Also decreased inflammation and fibrosis markers • Oral presentation EASL ILC Abstract AS074, Loomba et al., 2020 FASN in NASH • Primary human cell NASH model • Contains 4 liver cell types • Mimics human NASH by culture with glucose, fructose, fatty acids, insulin, LPS. • Developed and run by InSphero • TVB-2640 reduced markers of steatosis (cellular TGs), inflammation (IP-10) and fibrosis (Sirius Red) in the human primary cell LMT model. • Consistent with FASN inhibitor results in mouse NASH DIO and CCl4 models, previously published. • Decreased cellular TGs by TVB-2640 in the LMT model recapitulates reduced liver fat by TVB-2640 in the recent Ph2 FASCINATE-1 study in NASH patients. • Other NASH agents tested (Elafibranor, Obeticholic Acid, Resmetirom or Tropifexor) did not decrease cellular TGs in the LMT full NASH model as single agents, although decreases liver fat or steatosis have been shown in the clinic. These agents do not directly inhibit DNL, while ACC inhibitor GS-0976 had similar effect to TVB-2640 (not shown), consistent with direct DNL inhibition. The LMT full NASH model may not recapitulate the required mechanisms of altered lipid metabolism for other classes, and/or higher concentrations may be required. • Addition of TVB-2640 to the other NASH agents tested provided suppression of cellular TG in the LMT model in full NASH conditions. • Lipidomic profiling of supernatants by One Way Liver and assessment of fibrosis readouts such as ProC3 are ongoing. • The InSphero LMT model provides a useful tractable and non- invasive human model in NASH. LMTs were treated for 10 days with TVB-2640 in full NASH conditions (high sugars, FFA, insulin, LPS). Supernatants were tested for IP-10 levels on the Luminex platform (day 5). LMTs were lysed and assayed for TG levels using Glycerol/TG Glo kit (Promega) (day 10), corrected for basal TG levels. Results are mean +/-SEM of at least 4 LMT replicates per condition. *p<0.05 unpaired T test with Welch’s correction. For histology, a minimum of 16 LMTs (day 10) were pooled, processed and stained as indicated, 20x objective. TG; triglycerides, IP-10; interferon-inducible protein-10. LMTs were treated for 10 days with TVB-2460 and/or other compounds in full NASH conditions (high sugars, FFA, insulin, LPS). LMTs were lysed and assayed for TG levels by ELISA. The first purple bar is NASH conditions with vehicle only. Results are mean+/-SEM of at least 6 LMT replicates per condition, corrected for basal TG levels. Steatosis Cellular TGs Inflammation IP-10 Fibrosis Sirius Red Laboratory work conducted at InSphero. Thanks to the InSphero team for excellent collaboration (Sue Grepper, Manuela Bieri, Eva Thoma). 1 Syed-Abdul et al. 2020, Hepatology 72: 103. NASH condition (High sugars, insulin, FFA, LPS) In vitro culture for 10 days • Dose levels were selected based on published plasma levels in NASH clinical studies and/or in vitro cellular potency data • No cytotoxicity was observed with the combinations based on LDH assays on day 3, 5, 7, 10 • Elafibranor, Obeticholic Acid, Resmetirom or Tropifexor did not show meaningful inhibition of cellular TGs in NASH conditions. Combination with TVB-2640 provided suppression of cellular TG similar to that observed with TVB-2640 alone. • ALK5 inhibitor SB525334 and ASK1 inhibitor selonsertib were tested as single agents (not shown) and did not meaningfully suppress cellular triglyceride levels, but inhibited some inflammation and/or fibrosis parameters in this model • Similar combination effect with TVB-2640 was observed in a second model (InSphero’s DNL model) which lacks stellate cells and lacks exogenous FFA. Combination Study Cellular TGs Inhibitor Class Elafibranor (GFT505) PPAR a/d agonist Obeticholic acid / OCA (INT747) Bile acid analog FXR agonist Resmetirom (MGL-3196) THR-b agonist Tropifexor (LNJ452) Non bile acid FXR agonist TVB-2640 Acknowledgements Assay for triglycerides, cytokines, fibrosis markers, gene expression, histology. Marie O’Farrell, Douglas Buckley, George Kemble Elafibranor 1 uM Elafibranor 10 uM Control OCA 0.1 uM OCA 1 uM Tropifexor 0.2 nM Tropifexor 20 nM Resmetirom 50 nM Resmetirom 3 uM TVB-2640 0 300 600 900 1200 Triglycerides (pmol/MT) - + + - + + - + + - + + - + + - + + - + + - + + - + + below basal levels Kupffer cell Hepatic stellate cell Hepatocyte Hepatic endothelial cell Fibrosis Steatosis Inflammation 0 1000 2000 3000 IP-10 (pg/ml) TVB-2640 30 nM TVB-2640 300 nM TVB-2640 3 uM Vehicle -60% -58% -65% TVB-2640 30 nM TVB-2640 300 nM TVB-2640 3 uM Vehicle -57% -57% -64% 0 400 800 1200 1600 2000 Triglycerides (pmol/MT) * Sagimet Biosciences Inc, 155 Bovet Rd, San Mateo, CA 94402. Contact: [email protected]
Transcript of The FASN inhibitor TVB-2640 is efficacious in a new 3D ...
The FASN inhibitor TVB-2640 is efficacious in a new 3D human liver microtissue model of NASH
TVB-2640
Fibrosis
TVB-2640
SteatosisFASN
TVB-2
640
TVB-2640
Steatosis Inflammation
• Once-a-day oral small molecule• Potent with FASN EC50 approx. 50 nM• Excellent and predictable human PK profile• Inhibits hepatic DNL up to 90% in human 13C-acetate
tracer study1• First-in-class FASN inhibitor
Introduction
• De novo lipogenesis (DNL) is upregulated in NASH and drives steatosis and lipotoxicity
• Fatty acid synthase (FASN) is the last committed step in the DNL pathway and produces palmitate
• Palmitate is not only a building block for lipids, but a signaling molecule that activates the inflammasome and TLR pathways, impacting inflammation and fibrosis in NASH
Results in liver microtissue (LMT) model TVB-2640 inhibits three hallmarks of NASH; steatosis, inflammation and fibrosis Conclusions
TVB-2640 reduced liver fat in NASH patients in Ph2 FASCINATE-1 trial
• 61% patients treated with 50 mg TVB-2640 had ⩾ 30% reduction in liver fat by MRI-PDFF, vs 11% in placebo
• Also decreased inflammation and fibrosis markers • Oral presentation EASL ILC Abstract AS074, Loomba et
al., 2020
FASN in NASH
• Primary human cell NASH model• Contains 4 liver cell types• Mimics human NASH by culture with glucose,
fructose, fatty acids, insulin, LPS.• Developed and run by InSphero
• TVB-2640 reduced markers of steatosis (cellular TGs), inflammation (IP-10) and fibrosis (Sirius Red) in the human primary cell LMT model.
• Consistent with FASN inhibitor results in mouse NASH DIO and CCl4 models, previously published.
• Decreased cellular TGs by TVB-2640 in the LMT model recapitulates reduced liver fat by TVB-2640 in the recent Ph2 FASCINATE-1 study in NASH patients.
• Other NASH agents tested (Elafibranor, Obeticholic Acid, Resmetirom or Tropifexor) did not decrease cellular TGs in the LMT full NASH model as single agents, although decreases liver fat or steatosis have been shown in the clinic. These agents do not directly inhibit DNL, while ACC inhibitor GS-0976 had similar effect to TVB-2640 (not shown), consistent with direct DNL inhibition. The LMT full NASH model may not recapitulate the required mechanisms of altered lipid metabolism for other classes, and/or higher concentrations may be required.
• Addition of TVB-2640 to the other NASH agents tested provided suppression of cellular TG in the LMT model in full NASH conditions.
• Lipidomic profiling of supernatants by One Way Liver and assessment of fibrosis readouts such as ProC3 are ongoing.
• The InSphero LMT model provides a useful tractable and non-invasive human model in NASH.
LMTs were treated for 10 days with TVB-2640 in full NASH conditions (high sugars, FFA, insulin, LPS). Supernatants were tested for IP-10 levels on the Luminex platform (day 5). LMTs were lysed and assayed for TG levels using Glycerol/TG Glo kit (Promega) (day 10), corrected for basal TG levels. Results are mean +/-SEM of at least 4 LMT replicates per condition. *p<0.05 unpaired T test with Welch’s correction. For histology, a minimum of 16 LMTs (day 10) were pooled, processed and stained as indicated, 20x objective. TG; triglycerides, IP-10; interferon-inducible protein-10.
LMTs were treated for 10 days with TVB-2460 and/or other compounds in full NASH conditions (high sugars, FFA, insulin, LPS). LMTs were lysed and assayed for TG levels by ELISA. The first purple bar is NASH conditions with vehicle only. Results are mean+/-SEM of at least 6 LMT replicates per condition, corrected for basal TG levels.
SteatosisCellular TGs
InflammationIP-10
FibrosisSirius Red
Laboratory work conducted at InSphero. Thanks to the InSphero team for excellent collaboration (Sue Grepper, Manuela Bieri, Eva Thoma).
1Syed-Abdul et al. 2020, Hepatology 72: 103.
NASH condition (High sugars, insulin, FFA, LPS)
In vitro culture for 10 days
• Dose levels were selected based on published plasma levels in NASH clinical studies and/or in vitro cellular potency data
• No cytotoxicity was observed with the combinations based on LDH assays on day 3, 5, 7, 10
• Elafibranor, Obeticholic Acid, Resmetirom or Tropifexor did not show meaningful inhibition of cellular TGs in NASH conditions. Combination with TVB-2640 provided suppression of cellular TG similar to that observed with TVB-2640 alone.
• ALK5 inhibitor SB525334 and ASK1 inhibitor selonsertib were tested as single agents (not shown) and did not meaningfully suppress cellular triglyceride levels, but inhibited some inflammation and/or fibrosis parameters in this model
• Similar combination effect with TVB-2640 was observed in a second model (InSphero’s DNL model) which lacks stellate cells and lacks exogenous FFA.
Combination StudyCellular TGs
Inhibitor Class
Elafibranor (GFT505) PPAR a/d agonist
Obeticholic acid / OCA (INT747) Bile acid analog FXR agonist
Resmetirom (MGL-3196) THR-b agonist
Tropifexor (LNJ452) Non bile acid FXR agonist
TVB-2640
Acknowledgements
Assay for triglycerides, cytokines, fibrosis markers, gene expression, histology.
Marie O’Farrell, Douglas Buckley, George Kemble
Elafibranor1 uM
Elafibranor10 uM
Control OCA0.1 uM
OCA1 uM
Tropifexor0.2 nM
Tropifexor20 nM
Resmetirom50 nM
Resmetirom3 uM
TVB-2640
0
300
600
900
1200
Trig
lyce
rides
(pm
ol/M
T)
- + + - + + - + + - + + - + + - + + - + + - + + - + +
below basal levels
Kupffer cellHepatic stellate cell
Hepatocyte
Hepatic endothelial cell
Fibrosis
Steato
sis
Inflammation
0
1000
2000
3000
IP-1
0 (p
g/m
l)
TVB-264030 nM
TVB-2640300 nM
TVB-26403 uM
Vehicle
-60% -58%-65%
TVB-264030 nM
TVB-2640300 nM
TVB-26403 uM
Vehicle
-57% -57% -64%
0
400
800
1200
1600
2000Tr
igly
cerid
es (p
mol
/MT)
*
Sagimet Biosciences Inc, 155 Bovet Rd, San Mateo, CA 94402.
Contact: [email protected]
