1. • BDI-500 positively affects upstream mediators of adipose
tissue, systemic and hepatic inflammation and therefore
Normalizes metabolic profile
Reduces markers of CV risk
Improves glycemic control
Reduces markers of liver fibrosis
• BDI-500 significantly increases HNF4-α and therefore is an
ideal candidate for combination therapy with an FXR agonist
DBI-500 Significantly Increases Adipose Tissue Expression of PPAR-gamma and Adiponectin
in DIO Mice and Significantly Decreases Markers of Fibrosis in Two Murine NASH Models
Anton Leighton, MD
Dale Biotech
ABSTRACT SUMMARYOBJECTIVE MATERIALS AND METHODS
DISCUSSION
REFERENCES
COPYRIGHT OR FOOTER
• Review data supporting further development of DBI-500 as
Immunomodulatory treatment with potential to reverse
insulin resistance in obese T2DM patients
Oral treatment of obese T2DM patients with NASH
Introduction and Aims: The aims of these studies were to explore the scope of activity of
oral DBI-500, an atypical glycolipid, in a murine model of prediabetic type 2 diabetes and
obesity, and in two models of NASH.
Methods: Part 1. Male C57BL/6 mice (14 weeks old) that received a high fat diet were
divided into 3 groups (8 - 9 per group) and were either dosed with up to 144 mg/kg, 72 mg/kg or
matching placebo via once daily oral gavage for 7 days. Fasting glucose, body weight and gene
expression analyses were performed. Part 2. NASH was induced in male C57BL/6 mice using
a standard methionine/choline deficient diet. Concomitant with initiation of diet, 4 groups of 8
mice received either placebo, 80 or 40 mg/kg DBI-500 or 40 mg/kg + metformin every other day
for 4 weeks per gavage. Part 3. NASH was induced by diet in apolipoprotein-E-deficient mice
using western diet (WD). Dose groups of 5 dose groups of 8 mice were dose with DBI-500
(100, 50 or 25mg/kg, or 25mg+metformin).
Results: Part 1. Gene expression analysis of epididymal fat samples showed significant
increases in PPAR-gamma and adiponectin expression with concomitant significant reduction in
CD8+ expression. Blood glucose and body weight were reduced. Plasma levels of PAI-1 and
CD40L were also significantly reduced. Part 2. MCD mice showed significant and dose
dependent reduction in hepatic collagen. Part 3. In ApoE deficient mice, DBI-500 significantly
reduced expression of hepatic alpha-SMA and TGF-beta1, and significantly increased PPAR-
delta. BDI-500 also significantly increased HNF4-alpha.
Conclusions: DBI-500 is a potential once daily novel therapy for type two diabetes and
NASH that also significantly reduces markers of cardiovascular risk. DBI-500's capacity to
increase expression of HNF4-alpha strongly indicates potential as combination treatment with
FXR agonists.
BACKGROUND
• T2DM: patients increased incidence and severity of NASH
• Coexistence of NASH and T2DM results in a worse
metabolic profile and increased cardiovascular risk.
• New FDA draft guideline focuses on NASH with liver fibrosis
Fibrosis stage highly predictive for severe liver disease
• Regulatory pathway for T2DM is more straightforward than
that for NASH.
• BDI-500 mediates an upstream drivers of inflammation in the
complex process that drive NASH and T2DM, and is
therefore likely to present with enhanced efficacy in clinical
trials.
• Thomas AM, et al. Hepatocyte nuclear factor 4 alpha and farnesoid X receptor co-regulates gene transcription in mouse
livers on a genome-wide scale. Pharm Res. 2013.
• Baciu C, et al. Systematic integrative analysis of gene expression identifies HNF4A as the central gene in pathogenesis of
non-alcoholic steatohepatitis. PLoS One. 2017.
• McLaughlin T, et al. Role of innate and adaptive immunity in obesity-associated metabolic disease. J Clin Invest. 2017.
• Dooley S, ten Dijke P. TGF-β in progression of liver disease. Cell Tissue Res. 2012.
• Loria P, et al. Liver and diabetes. A vicious circle. Hepatol. Res. 2013.
• Nishikawa T, et al. Resetting the transcription factor network reverses terminal chronic hepatic failure. JCI 2015.
MATERIALS AND METHODS
BJECTIVE
• DBI-500 (Di-rhamnolipid C10-C10)
• Putative molecular target identified (CD1d)
• CD8 effector T cells infiltrate fatty tissue in obesity
• Influx of additional immune cells, incl. CD4 T cells and
macrophages
• Adipose inflammation
• Systemic inflammation
• Type 2 diabetes mellitus
• NASH
• NASH associated with insulin resistance and
accumulation of immune cells in liver as in adipose tissue
0
0.5
1
1.5
2
Placebo Mid Dose (MD) MD +
Metformin
High Dose
PercentArea
Fibrosis (% Collagen)
P<.001 (PBO vs HD)
0
0.5
1
1.5
2
2.5
3
Placebo Low Dose Mid Dose* Mid Dose +
Metformin*
High Dose*
FoldExpression
HNF4α Liver Expression (P<.05*)
Gene Expression in Visceral Adipose Tissue -
120mg/kg/d vs PBO; N=17
Gene Change P-Value Interpretation
CD8+ ↓71% <.01
Crucially involved in inflamm.
cascades in obese adipose
tissue and T2DM
CD4+ ↓63% <.05
Associated with obesity and
insulin resistance
PPAR-γ ↑93% <.05
Suppressed by inflammation;
Upregulates adiponectin
Adiponectin ↑65% <.05
Reduced in obese; insulin
resistance, dyslipidemia,
atherosclerosis. Biomarker
for in vivo PPAR-γ activation.
0
0.5
1
1.5
Placebo (n=8) 25mg (n=8) 50mg (n=8) 50mg +
Metformin
(n=8)
100mg (n=8)
α-SMA (liver) - Measure of Fibrosis
(P<.05 50mg vs Placebo)
0
0.5
1
1.5
Placebo (n=8) 25mg (n=8) 50mg (n=8) 50mg +
Metformin
(n=8)
100mg (n=8)
TGF-β1 (Liver) - Driver of Fibrosis
(P<.05 50mg vs Placebo)
• Increased α-SMA is
indicative of hepatic
stellate cell activation
and liver fibrosis
• TGF-β contributes to
hepatocyte death and
lipid accumulation to
promote NASH.
• HNF4-α expression
suppressed by TGF-β
• HNF4-α reduced in
NASH
• HNF4-α central regulator
of hepatic cell function in
interaction with FXR
Part 1: DIO mice received PBO, 60mg/kg or 120mg/kg
daily per oral gavage for 7 days
Part 2: MCD NASH Model. Mice received PBO, 40mg/kg
or 80mg/kg QAD/oral gavage for 28d; 8 mice/DG.
Part 3: Apo-E deficient mice received a Western diet and
either PBO, 25mg/kg, 50mg/kg, 50mg/kg + metformin, or
100mg/kg per oral gavage QAD for 28 days
• Established model
• Metformin added to
methionine-choline
deficient diet (MCD)
