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Long-acting GLP-1 analogue #2

Category
ID
Amount per vial
GLP-1
NNC0113-0217
1000 nmol

NNC0113-0217 (also known as semaglutide) is a GLP-1 analogue with 94% sequence homology to human GLP1 that binds and activates the GLP-1 receptor. Compared to native GLP-1, NNC0113-0217 have been designed to be long-acting in vivo. The main mechanism for the extended half-life is albumin binding. Furthermore, NNC0113-0217 has a delayed uptake from the subcutis. Also, NNC0113-0217 is fully stable against dipeptidyl-peptidase 4 (DPP-4).

GLP-1 action is mediated via a specific interaction with GLP-1 receptors, leading to an increase in cyclic adenosine monophosphate (cAMP). GLP-1 stimulates insulin secretion in a glucose-dependent manner. Simultaneously, GLP-1 lowers inappropriately high glucagon secretion, also in a glucose­dependent manner. Thus, when blood glucose is high, insulin secretion is stimulated and glucagon secretion is inhibited. Conversely, during hypoglycaemia GLP-1 diminishes insulin secretion and does not impair glucagon secretion. GLP-1 is a physiological regulator of appetite and food intake and the GLP-1 receptor is widely expressed in the brain.

Selected calculated properties for NNC0113-0217 and GLP-1(7-37)-OH are listed in the table.


PropertyNNC0113-0217GLP-1(7-37)-OH
MW (Da)4113.63355.7
pI (calculated)4.45.5
Sequence substitutions (compared to reference)8Aib, 26K(C18diacid-gGlu-2xOEG),34R-
Extinction coefficient (calculated, 280 nm)69906990

2D sketch of NNC0113-0217. NNC0113-0217 has a C18 diacid chain which is attached via a glutamic acid linker plus a hydrophilic spacer (2xOEG) to lysine at position 26. In addition, lysine is replaced with arginine at position 34. The fatty acid side chain enables reversible binding to serum albumin in the blood stream, which increases the half-life of the molecule. Furthermore, alanine at position 8 is substituted with 2-aminoisobutyric acid (Aib) to increase stability against DPP-IV cleavage.
2D sketch of human GLP-1 (7-37)-OH.

All in vitro data are based on in vitro assays using cloned human GLP-1 receptors expressed in baby hamster kidney cells. Since albumin binding is a key mechanism for the design of NNC0113-0217, the apparent affinity and potency will be very dependent on whether the assays contain albumin or not. See the Lau et al. 2015 reference listed in the reference section for further details on the experimental setup of the in vitro assays that have been used to generate the data in the table.


hGLP-1R binding (IC50, nM ± SEM)
Compound0% HSA2% HSA
GLP-1 (7-37)-OH0.19 ± 0.030.10 ± 0.02
NNC0113-02170.38 ± 0.06357 ± 98

hGLP-1R potency (EC50, nM ± SEM)
GLP-1 (7-37)-OH16.2 ± 0.9
NNC0113-02176.2 ± 0.6
hGLP-1R: human GLP-1 receptor; HSA: human serum albumin

The terminal half-life of NNC0113-0217 is estimated to be 8 h in the mouse, 11 hr in the rat, 28 h in the rabbit and 51 h in the monkey.

NNC0113-0217 has been tested in a range of diabetic and obese animal models. In normal male rats, the in vivo potency was estimated by dosing NNC0113-0217 subcutaneously (s.c.) followed by an i.v. glucose infusion 3 hrs later. NNC0113-0217 stimulated plasma insulin secretion and lowered blood glucose at a dose of 123 ug/kg.

In male diabetic db/db mice, upon single or repeated 4 week s.c. dosing, NNC0113-0217 lowers blood glucose dose-dependently and has a long duration of action. The ED50 for lowering of blood glucose (6 hours post dosing) is estimated to be 1.2 ug/kg for NNC0113-0217. The maximal effect on blood glucose lowering was obtained at 4-8 ug/kg for NNC0113-0217 in the 4 week study. The effect on body weight was maximal at a dose of 21 ug/kg.

The effects of NNC0113-0217 on body weight regulation have been evaluated in high fat diet obese (DIO) mice. Dosing of NNC0113-0217 for 18 days (0.15 mg/kg, s.c., daily) significantly lowered body weight.

When administering NNC0113-0217 to animals it is necessary to do dose titrations. A good dose for pharmacological experiments could be 120 ug/kg once daily titrated up from 12 ug/kg to 40 ug/kg to 120 ug/kg.

Human GLP-1(7-37)-OH (NNC0113-0007) is available as a reference compound to NNC0113-0217. Please indicate in the 'Purpose' field when you request NNC0113-0217 if you would like to have NNC0113-0007 included in your shipment.

Peptides and proteins have a tendency to adhere to glass and plastic surfaces. This may at low concentration impact the actual amount in solution. To minimize this unspecific adherence, adding detergents or inert proteins like e.g., ovalbumin or other serum albumins to the solution can minimize this phenomenon. In case albumins are added to peptide/protein solutions, ensure that the albumins are free of any proteases, but be aware that it will affect the apparent potency and affinity in in vitro assays in case a fatty acid is attached to the compound. For in vitro studies, NNC0113-0217 can be dissolved in 80/20% DMSO/MilliQ water (e.g. at a concentration of 300 uM). NNC0113-0217 can be dosed in vivo in a formulation vehicle containing 50mM sodium phosphate, 70mM sodium chloride, (0.007% polysorbate 20 if concentrations are so low that adsorption to vials may affect the concentration), pH 8.0. Formulations should be used fresh, but can be stored for up to one week refrigerated.

Lau J et al.
Discovery of the Once-Weekly Glucagon-Like Peptide-1 (GLP-1) Analogue Semaglutide
J Med Chem, 2015, 58, 7370-7380
Knudsen LB, Lau, J
The Discovery and Development of Liraglutide and Semaglutide
Front Endocrinol. 2019; 10:155