Small molecule HSL inhibitor

Category

HSL inhibitor

ID

NNC0076-0079

Amount per vial

20 mg

Hormone sensitive lipase (HSL) is an intracellular lipase found in most tissues. The lipase, that exists in a long and a short form, is capable of hydrolysing a variety of esters. The long form of HSL is expressed in steroidogenic tissues like testis, where it converts cholesteryl esters to free cholesterol for steroid hormone production. The short form is expressed in adipose tissue where it in concert with two other lipase, adipose triglyceride lipase (ATGL) and monoacylglycerol lipase (MGL), is involved in the hydrolysis triacylglycerols to free fatty acids and (FFAs) and glycerol. The main substrate for HSL in adipose tissue is diacylglycerol. The activity of HSL is regulated by insulin – it is high when circulating insulin levels are low and vice versa.

NNC0076-0079 is classified as a so-called substrate inhibitor. The HSL inhibitor NNC0076-0079 blocks the activity of HSL by interfering with its active site in a non-covalently fashion – the activation of HSL is regained ad the enzyme hydrolyses the ester bond of the inhibitor.

Category

HSL inhibitor

ID

NNC0076-0079

Amount per vial

20 mg

PropertyNNC0076-0079
MW [Da]256,69
Sum formulaC11 H13 Cl N2 O3

Figure 1

Chemical structure of NNC0076-0079. Morpholin-4-yl-carbamic acid 4-chloro-phenyl ester.
NNC0076-0079 selectively inhibits hormone sensitive lipase in vitro. The enzyme was preincubated with NNC0076-0079 (up to 50uM) for 30-70 min. at room temperature prior to addition of an emulsified fluorescent labeled triglyceride analogue substrate. Substrate and enzyme were allowed to incubate for 2 hours (37 degrees celcius). Hydrolysis was determined by fluorimetry.
EnzymeInhibition
(IC50, uM)
Hormone sensitive lipase (h-HSL)0,11
Lipoprotein lipase (LPL)>50
Hepatic lipase (HL)>50
Bile-salt stimulated lipase (BSSL)>50
Pancreatic lipase (PL)>50
NNC0076-0079 inhibits lipolysis in vivo. Study protocol for the data shown in the table: overnight fasted ob/ob mice, Sprague Dawley rats, high fat fed hamsters, and isoproterenol stimulated hamsters were treated orally with either vehicle or NNC0076-0079. The antilipolytic effects of NNC0076-0079 were evaluated in plasma samples collected 15 min – 5 hours after treatment using the biochemical markers glycerol and FFA. Compared to vehicle dosing, oral dosing of NNC0076-0079 in doses ≥ 30 mg/kg lowers circulating glycerol levels 30-75% relative to pre-dosing levels in the overnight fasted rat without access to food during the experiment. In this model, the glycerol lowering effect of NNC0076-0079 lasts for ≥ 3 hours. A similar effect pattern is seen in overnight-fasted mice.
SpeciesGlycerolFFA
ob/ob mouse 73%52%
Sprague Dawley rat59%36%
High fat fed hamster63%14%
Isoproterenol stimulated hamster51%41%
no reference compound is available
For in vitro experiments on cell supernatants and tissue cultures Stock solution: NNC0076-0079 is dissolved in DMSO and stored at 4 °C prior to use Test solution: 0.4% DMSO For in vivo experiments NNC0076-0079 can be formulated in the form of a suspension in 0,4 % Tween 80 (4 g/l) (BAKER 7394 batch 1931801868); 0,2 % carboxymethylcellulose sodium salt (2 g/l) (SIGMA C4888-500G batch SLCB7664). Make suspension vehicle as follows: 0.4 g Tween 80 and 0.2 g carboxymethylcellulose is dissolved in 100 ml MQ-H2O. An oral dose of 25 mg/kg and above gives rise to solid inhibition of HSL (drop in plasma glycerol) in the fasted rat.

de Jong JC et al.

Carbazates as potent inhibitors of hormone-sensitive lipase

Bioorg. Med. Chem. Lett., 14, 2004, 1741–1744

DOI PubMed

Alsted TJ et al.

Adipose triglyceride lipase in human skeletal muscle is upregulated by exercise training

Am J Physiol Endocrinol Metab 296: E445–E453, 2009

DOI PubMed

Wulf EM et al.

Antilipolytic effects of a novel hormone-sensitive lipase inhibitor (NNC 0076-0000-0079) in vitro and in vivo

American Diabetes Association 62nd Scientific Session, 2002, Poster 1379-P

DOI PubMed