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MGL - 3196, a β - Selective Thyroid Hormone Receptor (THR) Agonist , Demonstrates Metabolic , Anti - inflammatory and Anti - fibrotic Benefits in a Long - term High Fat Diet (HFD) Mouse NASH Model Rebecca Taub , John Franc, Martha Kelly; Madrigal Pharmaceuticals, Villanova, PA INTRODUCTION RESULTS The hepatic THR-β receptor mediates the beneficial effects of thyroid hormone on LDL-cholesterol and triglycerides, fatty liver and insulin sensitivity. Studies show that hepatic hypothyroidism is present in human NASH. MGL-3196 is a liver- directed, oral, selective THR-β agonist in Phase 2 for treatment of NASH. MGL-3196 is highly protein bound (>99%) with high liver uptake and low extrahepatic permeability. In C57Bl6 mice treated with 60% HFD for 38 weeks, MGL-3196 reduced cholesterol and ALT without the adverse effects of T3, which showed THRα mediated bone loss in the 3 week treatment. • These results confirm that prolonged treatment of mice with HFD generates a liver gene array profile consistent with activation of NASH and fibrosis pathways • MGL-3196 potentially normalizes hepatic function in HFD animals, including restoration of normal hepatic metabolic regulation without impacting tissues outside the liver • At human equivalent exposures MGL-3196 appears to reverse and prevent progression of lipid, inflammatory and fibrotic markers of NASH Mice on a HFD for 17 or 36 weeks were treated by daily oral gavage for 24 h or 24 days with 0.3-3 mg/kg/ day of MGL-3196. (Figure 2, 7 (panel R)) Mice on a HFD or normal chow (lean) or HFD plus oral admixture of 0.1, 0.3, 1.0 or 3 mg/kg/ day MGL- 3196 or 3 mg/kg/day rosiglitazone were treated for 25 weeks (Table 1, Figures 3-6, 7 Panel L). Mice were sacrificed and liver samples were processed and total RNA was extracted. Gene array studies were conducted using Illumina chips. At sacrifice blood samples were collected for clinical chemistry and lipid levels. MGL-3196 is specifically taken up into the liver by hepatic transporters (Figure 1). Metabolic effects of MGL-3196 treatment for 24 days (Figure 2; 7R) or 25 weeks (Table 1, Figures 3-7) included elimination of steatosis, normalization of liver size with no effect on overall body weight, improved insulin sensitivity, reduction in ALT (46%), free fatty acids (30%), and cholesterol (67%). Gene expression arrays confirmed regulation of known THR-β target genes such as Deiodinase-1, malic enzyme and FGF-21 (Figures 3). Metabolic genes associated with lipotoxicity such as acetyl CoA-carboxylase (ACC1) and 11-β Hydroxysteroid Dehydrogenase (HSD) were also normalized. In 25 week studies (Fig. 7, panel L) inflammatory (chemokine ligand 2 (CCL2), other chemokines) and major fibrosis associated genes (tissue inhibitor metalloproteinase (TIMP1), smooth muscle actin (SMA), connective tissue growth factor (CTGF) and collagen genes), were significantly increased in HFD as compared with lean controls and were normalized to the level of the lean control mice by treatment with human-relevant doses (0.3-1 mg/kg) MGL-3196 but not rosiglitazone which had a modest effect (Figure 7). In 24 day treatment of HFD mice (Fig. 7, panel R), a similar reduction of NASH inflammatory and fibrosis transcripts was observed. • J. Med Chem. 2014;57(10):3912-3923 • Thyroid hormone mediated autophagy and mitochondrial turnover in NAFLD, Sinha & Yen, Cell Biosci (2016) 6:46 CONCLUSIONS MATERIAL & METHODS DISCLOSURES Rebecca Taub, M.D., Chief Medical Officer, Executive Vice President R&D. Madrigal Pharmaceuticals. [email protected] Contact information Rebecca Taub - Management Position: Madrigal Pharmaceuticals John Franc – Madrigal Pharmaceuticals Martha Kelly - Madrigal Pharmaceuticals REFERENCES QR CODE Please retrieve your Poster QR Code during the Submission Process. The Inclusion of the QR code on your printed version will allow viewers to access your ePoster on the LiverLearning® To demonstrate the liver-directed pharmacologic effects of MGL-3196 in a mouse NASH model. AIM Table 1 MGL-3196 Liver Specific, Durable Fat Reduction, Metabolic Effects CFB, change from baseline; EE, energy expenditure; HFD, high fat diet; ITT, insulin tolerance test, ND: not done, value for 30 min shown, mice became hypoglycemic at 120 min (seizures); Rosi, rosiglitazone. Food intake and energy expenditure were percent difference relative to HFD control. No statistics were performed for food intake, because food intake was measured by group, not individual animal. #, p≤.001; ς, p≤.01; ¶, p≤.05 Assessment Chow Control HFD Control MGL-3196 Rosi 3mg/kg 0.1 mg/kg 0.3 mg/kg 1 mg/kg 3 mg/kg Insulin (ug/L) .27±.09 ς 3.24±3.21 2.46±2.58 ς 1.25±.60 1.09±1.5 .39±.26 ¶ .39±.22 ¶ Glucose (g/dL) 250±47.3 269±39 270±25.7 299±32.2 266±37.9 212±28.7 ς 270±27.3 Hgb A1C (%) 2.8±.23 # 3.2±.18 3.1±.22 3.1±.17 3.0±.32 2.8±.27 ς 3.2±.24 ITT (glucose CFB) @120 ‘ -14.3±9.9 -8.1±10.8 -17.7±18.0 -32.8±22.3 ς -42.4±12.1 # -73.2±10.9 # ND -31.0±19.8 ς ALT (U/L) 53±12 85±57 55±37 46±11 ¶ 54±50 58±22 49±19 Cholesterol (mg/dL) 87±11.8 # 219±26.4 170±45.5 ς 117±19.8 # 96±20.2 # 72±23.9 # 147±13.5 # Triglycerides (mg/dL) 132±34.8 127±49.2 118±45.4 91±31.8 101±50.2 83±24.5 ¶ 104±36.9 FFA (mmol/L) 2.15±.60 1.91±.25 1.80±.34 1.33±.36 # 1.76±.52 1.62±.41 1.36±.23 # Body Weight (g) 30.1±2.5 # 49.7±2.1 46.8±9.8 45.6±5.5 ς 41.0±4.9 # 37.0±6.1 # 51.0±7.8 Food Intake (% Control) 37.5 2.2 19.6 27.7 30.6 -1.4 EE (% control) -10.64 ς 6.58 5.67 ¶ 16.9 # 26.9 # 4.24 Liver Weight (%BW) 4.7±.7 4.4±.7 3.8±.7 3.1±.2# 3.3±.16# 3.3±.4 # 3.4±.3 ς Fat Mass (g) 5.2±.5 20.8±3.9 20.1±3.4 20.0±5.1 20.1±3.8 13.0±1.8 ς 28.7±3.7 # Lean Mass (g) 26.3±1.1 27.4±2.0 26.2±2 26.3±1.7 25.7±1.0 24.6±2.2 ¶ 25.0±.8 ¶ Figure 6. Liver-specific Fat Reduction 0 0.2 0.4 0.6 0.8 1 1.2 1.4 1.6 Body Weight (g) Liver Weight (% Body Weight) Liver weight absolute Fat Mass (g) Lean Mass (g) Energy expenditure Chow HFD MGL-3196.1mg/kg MGL-3196 .3mg/kg MGL-3196 1mg/kg MGL-3196 3mg/kg Rosi 3mg/kg • 0.3 mg/kg exposure equivalent to human 80 mg dose • Liver weight normalized at 0.3 mg/kg with no effect on body weight-animals remain obese Figure 5. Control MGL-3196 400X 100X Liver Histology in 36 Week HFD Mice Treated with MGL- 3196 • 24 day treatment with MGL-3196 normalized liver histology at all doses (.3-10 mg/kg) • Consistent with THR-β effects to increase fat oxidation • No detectable steatosis, fibrosis or inflammation in MGL-3196 treated livers Figure 1. Radiographic Tissue Distribution- Rats Rat 14 C ADME study (Ref 1) • MGL-3196 uptake in liver • Low to undetectable level in heart, bone, brain • Primarily fecal elimination via biliary excretion Figure 4. Normalization of Hepatic Gene Expression • 25 week treatment with HFD changes the hepatic expression of a large number of gene transcripts, including both increased (red) and decreased (blue) expression • Treatment with MGL-3196 normalizes hepatic gene expression as well as hepatic architecture and size HFD 0.1 0.3 1 3 Rosi MGL-3196 (mg/kg) Figure 2. Improvement in Liver Health 0 50 100 150 200 250 300 IU/L MGL-3196--ALT Control MGL-3196 .3mg/kg MGL-3196 1mg/kg MGL-3196 3mg/kg MGL-3196 10mg/kg *** *** *** *** 0 50 100 150 200 250 300 IU/L T3 Treatment--ALT Control T3 10ug/kg T3 30ug/kg T3 100ug/kg *** *** ** 0 1 2 3 4 (g) T3 Treatment--Liver Size 0 20 40 60 80 100 120 0 60 120 % Time 0 Glucose Time, (min) Insulin Tolerance Test (0.5 U/kg insulin) Control MGL-3196 .3mg/kg MGL-3196 1mg/kg MGL-3196 3 mg/kg Rosiglitazone: 10 mg/kg 0 10 20 30 40 50 60 70 Triglycerides mg/g * ** ** Liver Triglycerides 0 1 2 3 4 (g) MGL-3196 --Liver Size ** ** ** ** p<.05*, p<.01**, P<.001*** • 24 d treatment MGL-3196 and T3 reduce ALT • MGL-3196 also reduces liver size and liver triglycerides Figure 7. MGL-3196 Reduces NASH & Fibrosis and Increases Mitophagy Pathway Genes “HFD”, lane 1 mean HFD gene expression normalized to mean Lean; Lanes (2-7) mean gene expression normalized to mean of HFD; “Rosi” (rosiglitazone, 3 mg/kg, 25 weeks); lanes 8-10, treatment 24 d MGL-3196 or Rosi (10 mg/kg), normalized to HFD Control; Red, higher expression; blue decreased expression Inflammatory HFD Lean 0.1 0.3 1 3 Rosi 0.3 1 3 Rosi MCP-1/CCL2 MIP-2α/CXCL2 MIP-2ß/CXLCL3 A20/TNFaip3 CRP Annexin 2 SAA1 Fibrosis TGFbeta Osteopontin Hyaluron syn Collagen 1 Galectin-3 TIMP1 Collagen 4a2 SMA Collagen 4a1 CTGF Keratin 18 Collagen 3 Galectin-1 Mitophagy-Ulk-1 25 week treatment MGL-3196 (mg/kg) MGL-3196 24 day treatment • Normalization of NASH-related transcripts to level of lean control • Stimulation of mitophagy via ULK1 induction by THR-β leads to mitochondrial biogenesis which protects against NASH-related ROS, lipotoxicity Figure 3. DIO1, deiodinase 1;TBG, thyroid binding globulin (SerpinA); mGPDH, mitochondrial glycerol-3-phosphate dehydrogenase 0 1 2 3 4 5 6 7 8 Dio1 TBG Malic Enzyme mGPDH ACC1 11BHSD1 Lean Control DIO Control MGL-3196 .1mg/kg MGL-3196 .3mg/kg MGL-3196 1mg/kg MGL-3196 3mg/kg Rosi 3mg/kg 0 2 4 6 8 10 12 14 16 18 20 FGF21 MGL-3196 Regulates Metabolic and THR-β Responsive Genes
Transcript of QR CODE MGL-3196, a β-Selective Thyroid Hormone ......MGL-3196, a β-Selective Thyroid Hormone...
MGL-3196,aβ-Selective Thyroid HormoneReceptor (THR)Agonist,DemonstratesMetabolic,Anti-inflammatory andAnti-fibrotic Benefits inaLong-term HighFatDiet(HFD)MouseNASHModelRebeccaTaub,JohnFranc,MarthaKelly;MadrigalPharmaceuticals,Villanova,PA
INTRODUCTION RESULTSThe hepatic THR-β receptormediates the beneficial effects ofthyroid hormone on LDL-cholesteroland triglycerides, fatty liver andinsulin sensitivity. Studies show thathepatic hypothyroidism is present inhuman NASH. MGL-3196 is a liver-directed, oral, selective THR-βagonist in Phase 2 for treatment ofNASH. MGL-3196 is highly proteinbound (>99%) with high liver uptakeand low extrahepatic permeability.In C57Bl6 mice treated with 60%HFD for 38 weeks, MGL-3196reduced cholesterol and ALT withoutthe adverse effects of T3, whichshowed THRα mediated bone loss inthe 3 week treatment.
• These results confirm thatprolonged treatment of mice withHFD generates a liver gene arrayprofile consistent with activationof NASH and fibrosis pathways
• MGL-3196 potentially normalizeshepatic function in HFD animals,including restoration of normalhepatic metabolic regulationwithout impacting tissues outsidethe liver
• At human equivalent exposuresMGL-3196 appears to reverse andprevent progression of lipid,inflammatory and fibrotic markersof NASH
Mice on a HFD for 17 or 36 weekswere treated by daily oral gavage for24 h or 24 days with 0.3-3 mg/kg/day of MGL-3196. (Figure 2, 7 (panelR)) Mice on a HFD or normal chow(lean) or HFD plus oral admixture of0.1, 0.3, 1.0 or 3 mg/kg/ day MGL-3196 or 3 mg/kg/day rosiglitazonewere treated for 25 weeks (Table 1,Figures 3-6, 7 Panel L). Mice weresacrificed and liver samples wereprocessed and total RNA wasextracted. Gene array studies wereconducted using Illumina chips. Atsacrifice blood samples werecollected for clinical chemistry andlipid levels.
MGL-3196 is specifically taken up into the liver by hepatic transporters (Figure 1). Metabolic effects of MGL-3196 treatment for 24 days (Figure 2; 7R) or 25 weeks (Table 1, Figures 3-7) included elimination of steatosis,normalization of liver size with no effect on overall body weight, improved insulin sensitivity, reduction in ALT (46%), free fatty acids (30%), and cholesterol (67%). Gene expression arrays confirmed regulation of known THR-βtarget genes such as Deiodinase-1, malic enzyme and FGF-21 (Figures 3). Metabolic genes associated with lipotoxicity such as acetyl CoA-carboxylase (ACC1) and 11-β Hydroxysteroid Dehydrogenase (HSD) were alsonormalized. In 25 week studies (Fig. 7, panel L) inflammatory (chemokine ligand 2 (CCL2), other chemokines) and major fibrosis associated genes (tissue inhibitor metalloproteinase (TIMP1), smooth muscle actin (SMA),connective tissue growth factor (CTGF) and collagen genes), were significantly increased in HFD as compared with lean controls and were normalized to the level of the lean control mice by treatment with human-relevantdoses (0.3-1 mg/kg) MGL-3196 but not rosiglitazone which had a modest effect (Figure 7). In 24 day treatment of HFD mice (Fig. 7, panel R), a similar reduction of NASH inflammatory and fibrosis transcripts was observed.
• J. Med Chem. 2014;57(10):3912-3923• Thyroid hormone mediated
autophagy and mitochondrialturnover in NAFLD, Sinha & Yen, CellBiosci (2016) 6:46
CONCLUSIONS
MATERIAL&METHODS
DISCLOSURES
Rebecca Taub, M.D., Chief Medical Officer, Executive Vice PresidentR&D. Madrigal Pharmaceuticals. [email protected]
RebeccaTaub- ManagementPosition:MadrigalPharmaceuticals
JohnFranc– MadrigalPharmaceuticals
MarthaKelly- MadrigalPharmaceuticals
REFERENCES
QRCODEPleaseretrieveyourPosterQRCodeduringtheSubmission
Process.
TheInclusionoftheQRcodeonyourprintedversionwillallowviewerstoaccessyourePoster
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To demonstrate the liver-directedpharmacologic effects of MGL-3196in a mouse NASH model.
AIM
Table 1 MGL-3196 Liver Specific, Durable Fat Reduction, Metabolic Effects
CFB, change from baseline; EE, energy expenditure; HFD, high fat diet; ITT, insulin tolerance test, ND: not done, value for 30 min shown, mice became hypoglycemic at 120 min (seizures); Rosi, rosiglitazone. Food intake and energy expenditure were percent difference relative to HFD control. No statistics were performed for food intake, because food intake was measured by group, not individual animal. #, p≤.001; ς, p≤.01; ¶, p≤.05
Assessment Chow Control
HFD Control
MGL-3196 Rosi3mg/kg0.1 mg/kg 0.3 mg/kg 1 mg/kg 3 mg/kg
Insulin (ug/L) .27±.09ς 3.24±3.21 2.46±2.58ς 1.25±.60 1.09±1.5 .39±.26¶ .39±.22¶
Glucose (g/dL) 250±47.3 269±39 270±25.7 299±32.2 266±37.9 212±28.7ς 270±27.3
Hgb A1C (%) 2.8±.23# 3.2±.18 3.1±.22 3.1±.17 3.0±.32 2.8±.27ς 3.2±.24
ITT (glucose CFB) @120 ‘ -14.3±9.9 -8.1±10.8 -17.7±18.0 -32.8±22.3ς -42.4±12.1# -73.2±10.9#
ND -31.0±19.8ς
ALT (U/L) 53±12 85±57 55±37 46±11¶ 54±50 58±22 49±19
Cholesterol (mg/dL) 87±11.8# 219±26.4 170±45.5ς 117±19.8# 96±20.2# 72±23.9# 147±13.5#
Triglycerides (mg/dL) 132±34.8 127±49.2 118±45.4 91±31.8 101±50.2 83±24.5¶ 104±36.9
FFA (mmol/L) 2.15±.60 1.91±.25 1.80±.34 1.33±.36# 1.76±.52 1.62±.41 1.36±.23#
Body Weight (g) 30.1±2.5# 49.7±2.1 46.8±9.8 45.6±5.5ς 41.0±4.9# 37.0±6.1# 51.0±7.8
Food Intake (% Control) 37.5 2.2 19.6 27.7 30.6 -1.4
EE (% control) -10.64ς 6.58 5.67¶ 16.9# 26.9# 4.24
Liver Weight (%BW) 4.7±.7 4.4±.7 3.8±.7 3.1±.2# 3.3±.16# 3.3±.4# 3.4±.3ς
Fat Mass (g) 5.2±.5 20.8±3.9 20.1±3.4 20.0±5.1 20.1±3.8 13.0±1.8ς 28.7±3.7#
Lean Mass (g) 26.3±1.1 27.4±2.0 26.2±2 26.3±1.7 25.7±1.0 24.6±2.2¶ 25.0±.8¶
Figure 6. Liver-specific Fat Reduction
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• 0.3mg/kgexposureequivalenttohuman80mgdose• Liverweightnormalizedat0.3mg/kgwithnoeffecton
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Liver Histology in 36 WeekHFD Mice Treated with MGL-3196
• 24daytreatmentwithMGL-3196normalizedliverhistologyatalldoses(.3-10mg/kg)
• ConsistentwithTHR-βeffectstoincreasefatoxidation• Nodetectablesteatosis,fibrosisorinflammationinMGL-3196treatedlivers
Figure 1. RadiographicTissueDistribution-Rats
Rat14CADMEstudy(Ref1) • MGL-3196uptakeinliver
• Lowtoundetectablelevelinheart,bone,brain
• Primarilyfecaleliminationviabiliaryexcretion
Figure 4. Normalization ofHepatic Gene Expression• 25weektreatmentwithHFDchangesthe
hepaticexpressionofalargenumberofgenetranscripts,includingbothincreased(red)anddecreased(blue)expression
• TreatmentwithMGL-3196normalizeshepaticgeneexpressionaswellashepaticarchitectureandsize
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Figure 2. Improvement in Liver Health
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• 24dtreatmentMGL-3196andT3reduceALT• MGL-3196alsoreducesliversizeandlivertriglycerides
Figure 7. MGL-3196 Reduces NASH & Fibrosisand Increases Mitophagy Pathway Genes
“HFD”, lane 1 mean HFD gene expression normalized to mean Lean; Lanes (2-7) mean gene expression normalized to mean of HFD; “Rosi” (rosiglitazone, 3 mg/kg, 25 weeks); lanes 8-10, treatment 24 d MGL-3196 or Rosi (10 mg/kg), normalized to HFD Control; Red, higher expression; blue decreased expression
Inflammatory HFDLean 0.1 0.3 1 3 Rosi 0.3 1 3 RosiMCP-1/CCL2MIP-2α/CXCL2MIP-2ß/CXLCL3A20/TNFaip3CRPAnnexin 2SAA1FibrosisTGFbetaOsteopontinHyaluron synCollagen 1Galectin-3TIMP1Collagen 4a2SMACollagen 4a1CTGFKeratin 18Collagen 3Galectin-1Mitophagy-Ulk-1
25weektreatmentMGL-3196(mg/kg) MGL-3196
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• NormalizationofNASH-relatedtranscriptstolevelofleancontrol
• StimulationofmitophagyviaULK1inductionbyTHR-βleadstomitochondrialbiogenesiswhichprotectsagainstNASH-relatedROS,lipotoxicity
Figure 3.
DIO1,deiodinase1;TBG,thyroidbindingglobulin(SerpinA);mGPDH,mitochondrialglycerol-3-phosphatedehydrogenase
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