Table 4.

Key animal studies regarding the impact of ipragliflozin, remogliflozin, tofogliflozin and luseogliflozin on NAFLD/NASH/HCC.

Study/Reference	Animal Model Dose & Duration	Effect on Body Weight &Body Composition	Effect on Laboratory Values	Mechanism of Action	Effect on Insulin Sensitivity & Glucose Homeostasis	NAFLD Activity Score (NAS) & Fibrosis/Steatosis
Ipragliflozin
Tahara, A. & Takasu, T., 2020 [100]	KK-Ay mice on HFD 0.1, 0.3, 1 and 3 mg/kg/day Alone or with Metformin 4 (w)	⬇ BW weight (1 & 3 mg) ⬇ Liver weight (0.3, 1 & 3 mg)	⬇ TG (0.3, 1 & 3 mg) ⬇ TC (1 & 3 mg) ⬇ AST (1 & 3 mg) ⬇ ALT (0.3, 1 & 3 mg)	⬇ Inflammatory Markers [serum TNF-α, IL-6, MCP-1 and CRP (1 and 3mg); Liver TNF-α (3 mg) and IL-6, MCP-1 and CRP (1 & 3 mg)] ⬇ Serum and hepatic oxidative stress markers [TBARS and protein carbonyl (1 & 3 mg)]	Improve glucose intolerance Improved Insulin resistance Improved hyperlipidemia	⬇ Hepatic TG, TC (1 & 3 mg) ⬇ Hepatic Hyperthrophy (1 & 3 mg) ⬇ Hepatic Inflammation (1 & 3 mg) ⬇ Hepatic fibrosis & steatosis (3 mg)
Tahara, A., et al., 2019 [113]	KK-Ay mice on HFD 0.1, 0.3, 1 and 3 mg/kg/day Alone or with Pioglitazone 4 (w)	⬇ BW weight (1 & 3 mg) ⬇ Liver weight (0.3, 1 & 3 mg)	⬇ TC (0.3, 1 & 3 mg) ⬇ TG (1 & 3 mg) ⬇ AST (1 & 3 mg) ⬇ ALT (1 & 3 mg)	⬇ Genes involved in regulation of insulin sensitivity (Plasma adipocytokines, Leptin & FGF-21) ⬇ Inflammatory Markers [serum TNF-α, IL-6, MCP-1 and CRP (1 and 3 mg); Liver TNF-α (3 mg) and IL-6, MCP-1 and CRP (1 and 3 mg)] ⬇ Serum and hepatic oxidative stress markers [TBARS and protein carbonyl (1 & 3 mg)]	⬇ Plasma glucose and insulin levels (0.3, 1 and 3 mg)	⬇ Hepatic TG (0.3, 1 & 3 mg) ⬇ Hepatic TC (1 & 3 mg) ⬇ Hepatic Hyperthrophy (1 & 3 mg) ⬇ Hepatic Inflammation (3 mg) ⬇ Hepatic fibrosis (3 mg)
Komiya, Ch, et al., 2016 [112]	ob/ob and WT mice on HFD 11 mg/kg/day 4 (w)	⬇ Hyperphagia ⬇ BW weight ⬇ Liver weight	⬇ ALT ⬇ TG ⬇ Plasma glucagon	⬇ Lipogenic markers genes (SREBP1, Fasn, Acc1, Scd1) ⬇ Gluconeogenic markers (Pck1) ⬇ Inflammatory markers (F4/80, Cd11c)	Improved Insulin resistance Improved fasting glucose levels	⬇ Hepatic TG ⬇ Hepatic lipid ⬇ Hepatic steatosis
Honda, Y., et al., 2016 [114]	C57BL/6J male mice on AMLN diet 40 mg/kg/day 8 (w)	No effect on BW ⬇ Liver weight	⬇ ALT, AST ⬇ FFA	⬆ β-oxidation (PPAR-α, CPT1, MTTP) ⬇ Hepatocytes apoptosis (TUNEL) ⬇ Lipogenic markers genes (SREBP1 & Acc1)	Improved Insulin resistance	⬇ Hepatic TG & FFA ⬇ Hepatic fibrosis ⬇ Hepatocyte ballooning ⬇ Lobular inflammation ⬇ NAS
Hayashizaki-Someya, Y., et al., 2015 [144]	Male Wistar rats on CDAA diet 0.3 and 3 mg/kg/day 5 (w)	⬇ BW weight (3 mg)	No effect on ALT, AST	-	No effect on fasting blood glucose levels	⬇ Hepatic TG (3 mg) ⬇ Hepatic lipid droplet size ⬇ Hepatic fibrosis (0.3 & 3 mg) ⬇ Hepatic HP
Yoshioka, N., et al., 2021 [126]	Mc4r KO mice on HFD and injected with single dose of diethylnitrosamine 5 mg/kg/day 12 (w)	⬇ BW weight ⬇ Liver weight	⬇ ALT, AST ⬇ LDH	⬇ Lipogenic markers genes (Fasn in non-tumor) ⬇ Fibrosis markers (Emr1, Itgax in non-tumor) ⬇ Cell senescence markers (Cxcl1 in tumor lesion; p21, Cxcl1, MMp12, mmp13 in non-tumor) ⬆ β-oxidation (PPAR-α in tumor lesion; PPAR-α, CPT1, PGC1 in non-tumor) ⬇ Cell apoptosis (Bax and Pcna)	⬇ Plasma glucose & insulin levels	⬇ Hepatic TG ⬇ Lobular inflammation ⬇ Hepatocyte ballooning ⬇ NAS ⬇ Hepatic steatosis & fibrosis ⬇ Hepatic tumor number & size
(Remogliflozin)
Nakano, S., et al., 2015 [134]	C57BL/6J mice on HFD32 13.2 ± 2.2 and 33.9 ± 2.0 mg/kg/day 4 (w)	⬇ Liver weight ⬇ Liver/BW	⬇ ALT & AST	⬇ Inflammatory markers [Hepatic TNF-α (13.2 mg), hepatic MCP-1 (13.2 and 33.9 mg)] ⬇ Oxidative stress (serum and hepatic TBARS)	Improved non fasting glucose levels	⬇ Hepatic TG ⬇ Hepatic fibrosis
Tofogliflozin
Obara, K., et al., 2017 [89]	db/db mice on HFD and injected with single dose of diethylnitrosamine 1 and 10 mg/kg/day 14 (w)	⬇ Liver weight (10 mg)	⬇ ALT (10 mg) ⬇ FFA (1 & 10 mg)	⬇ Inflammatory markers (10 mg) (F4/80)	⬇ Plasma glucose levels Improved insulin insensitivity	⬇ Foci of cellular alteration (10 mg) ⬇ Hepatic pre-neoplastic lesions (10 mg) ⬇ Hepatocyte balooning (10 mg) ⬇ Hepatic steatosis (10 mg) ⬇ NAS (1 & 10 mg)
Luseogliflozin
Qiang, Sh, et al., 2015 [142]	C57BL/6 mice injected with STZ on HFDT Mixing in food at 0.1%. w/w food 8 (w)	⬇ Liver weight	⬇ ALT ⬆ TG, NEFA	⬇ Hepatic fibrosis markers (collagen1a1, collagen1a2, TGF, SMA, TIMP1) ⬇ Inflammatory markers (MCP-1, IL1, IL-12, IL-6, f4/80)	⬇ Plasma glucose levels	⬇ Hepatic TC, TG &NEFA

Abbreviations: W: week, BW: Body Weight, STZ: streptozotocin, WT: Wild Type, Gla:insulin glargine, ALT: Alanine aminotransferase, AST: Aspartate aminotransferase, TG: Triglycerides, TC: Total cholesterol, FFA: Free fatty acids, GA: glycated albumin, STAT3: Signal Transducer And Activator Of Transcription 3, ROR: RAR Related Orphan Receptor, Bax: BCL2 Associated X, IRE: Inositol-requiring enzyme 1, -Xbp1: X-box binding protein 1, ATF4: Activating transcription factor 4, CHOP: C/EBP homologous protein, ATF-6:Activating transcription factor 6, ChREBP: Carbohydrate response element binding protein, SREBP1: Sterol regulatory element-binding transcription factor 1, Scd1: Stearoyl-CoA desaturase1, ACC1: Acetyl-CoAcarboxylase 1, ACOX: Peroxisomal acyl-CoA oxidase1 Fasn: Fatty acid synthase, IL: Interleukin, TNF-α: Tumor necrosis factor, NFκΒ: Nuclear factor kappa-light-chain-enhancer of activated B cells, MCP-1:Monocyte chemoattractant protein-1, CRP:C-reactive protein, PPAR-α: Peroxisome proliferator activated receptor alpha, CPT1: Carnitine palmitoyltransferase 1, PGC1: PPARγ coactivator 1, TOS: Total Oxidant Status, TAS: Total antioxidant status, p21: cyclin-dependent kinase inhibitor, Cxcl: Chemokine (C-X-C motif) ligand, MMP: Matrix metalloproteinases, TIMP: Tissue inhibitors of matrix metalloproteinases, CYP: Cytochromes P450, MDA: Malondialdehyde.