An approach to

half-life extension
Paper by: and
Kim at al.
Presentation by:
activity preservation
Ghufran Ayman. of
SEMINAR OF
BIOTECHNOLOGY AND
an anti- diabetic peptide drug
MOLECULAR BIOLOGY. based on
genetic fusion with an albumin
binding aptide.
Introduction

Exenatide:
Is a peptide drug,
Is used for the treatment of Type 2
Diabetes Mellitus (T2DM),
Has a half life of 1.5-4 hours.
Background

• Type 2 diabetes is characterised by

impaired insulin secretion, caused by
decreased secretion of glucagon-like
peptide-1 (GLP-1).
• Exenatide is a GLP-1 mimetic
peptide, used as an adjunctive
therapy to improve glycaemic control
in T2DM patients.
• Aptides are a novel class of structure-
constrained peptides containing a
randomizable binding region and a
constant β-hairpin scaffold.

Figure 1: action of GLP-1.

The authors
propose:

• Novel long-acting fusion

peptide comprising of
Exenatide and a Human
Serum Albumin-Binding
Aptide (APTHSA).

Figure 2: Exenatide-Aptide fusion.

 • Phage display screening for HSA-binding
Studying aptides
the
aptide • Specificity assessment of APTHSA
Methods • Measurement of APTHSA affinity
• Preparation of an exenatide-APTHSA fusion
& peptide
• In vitro biological activity of exenatide-APTHSA
Results • In vivo pharmacokinetics of exenatide-APTHSA
Studying • Anti-hyperglycemic efficacy of exenatide-
the APTHSA in fasted db/db mice
effect of • Hypoglycemic efficacy of exenatide-APTHSA in
Drug- non-fasted db/db mice
Aptide
fusion • Statistical analysis
 Phage display screening.

Studying the
aptide

Figure 3: phage display screening.

The result of phage
display screening is
a large amount of
the antibody
bacteriophage that
has the human-
serum-albumin-
binding aptide.

Figure 4: Phage antibody with human

serum albumin binding aptide.
 Phage ELISA.

Studying the Aptide

aptide
 The result of
phage ELISA is
the Aptide
bacteriophage
produced is
highly specific to
Human serum
albumin.
 Recombinant DNA technology.

Fusion Gene
PCR

Preparation of
the Drug-Aptide
fusion
Prokaryotic cell

Exenatide-Aptide
recombinant
plasmid
 Preparation of
the Drug-Aptide
fusion
 In Vitro In Vivo

Pancreatic cell line. Subcutaneous administration

of exenatide and exenatide-
APT HSA that were mixed with
Treatment with HSA in mice.
exenatide-APT HSA fusion.
Obtainment of blood samples.
Studying the
effects of Drug- Measurement of insulin
Aptide fusion secretion Measurement of concentration
of exenatide or exenatide-APT HSA

Estimation of Pharmacokinetic
parameters, anti-hyperglycemic efficacies
and hypoglycemic efficacies.
The result of In vitro
studies is that
exenatide-APTHSA
retained nearly 90%
of the biological
activity of the
original exenatide
 The result of In vivo
Pharmacokinetic
analysis is that
exenatide-APTHSA
showed slower
clearance and longer
half life than
exenatide.
The result of In vivo
Anti-hyperglycemic
activity assessment
is that exenatide-
APTHSA retains
biological activity
similar to that of
exenatide.
The result of In vivo
Hypoglycemic effect
assessment is that
exenatide-APTHSA
showed increased
duration of action.
Conclusion

The authors succeeded in developing a

novel fusion peptide between
exenatide and a HSA-binding aptide.

The exenatide-APTHSA:
-preserved the biological activity of exenatide in vitro as well as in vivo
-and showed significantly improved pharmacokinetic characteristics
compared with exenatide.
• The strategy described here could be applied to other
important peptide- or protein-based biotherapeutics.
• The affinity of the present APTHSA for HSA will need to be
significantly improved to further increase blood circulation
time,
• Safety profile should be studied.

Future direction