Long-Acting PASylated Leptin Ameliorates Obesity by Promoting Satiety and Preventing Hypometabolism in Leptin-Deficient Lep(ob/ob) Mice.
Body weight loss of Lep(ob/ob) mice in response to leptin is larger than expected from the reduction in energy intake alone, suggesting a thermogenic action of unknown magnitude. We exploited the superior pharmacological properties of a novel long-acting leptin prepared via PASylation to study the contribution of its anorexigenic and thermogenic effects. PASylation, the genetic fusion of leptin with a conformationally disordered polypeptide comprising 600 Pro/Ala/Ser (PAS) residues, provides a superior way to increase the hydrodynamic volume of the fusion protein, thus retarding kidney filtration and extending plasma half-life. Here a single PAS(600)-leptin injection (300 pmol/g) resulted in a maximal weight reduction of 21% 6 days after application. The negative energy balance of 300 kJ/(4 d) was driven by a decrease in energy intake, whereas energy expenditure remained stable. Mice that were food restricted to the same extent showed an energy deficit of only 220 kJ/(4 d) owing to recurring torpor bouts. Therefore, the anorexigenic effect of PAS(600)-leptin contributes 75% to weight loss, whereas the thermogenic action accounts for 25% by preventing hypometabolism. In a second experiment, just four injections of PAS(600)-leptin (100 pmol/g) administered in 5- to 6-day intervals rectified the Lep(ob/ob) phenotype. In total, 16 nmol of PAS(600)-leptin per mouse triggered a weight loss of 43% within 20 days and normalized hypothermia and glucose homeostasis as well as hepatic steatosis. The beneficial properties of PAS(600)-leptin are substantiated by a comparison with previous studies in which approximately 400 nmol (∼25-fold) unmodified leptin was mandatory to achieve similar improvements.
Lehrstuhl für Molekulare Ernährungsmedizin (F.B., A.B., N.R., S.M., M.K.), Else Kröner-Fresenius Center and ZIEL-Research Center for Nutrition and Food Science, and Munich Center for Integrated Protein Science and Lehrstuhl für Biologische Chemie (V.M., M.S., A.S.), Technische Universität München, 85350 Freising-Weihenstephan, Germany; and XL-protein GmbH (A.S.), 85354 Freising, Germany.
Florian
F
Bolze
Lehrstuhl für Molekulare Ernährungsmedizin (F.B., A.B., N.R., S.M., M.K.), Else Kröner-Fresenius Center and ZIEL-Research Center for Nutrition and Food Science, and Munich Center for Integrated Protein Science and Lehrstuhl für Biologische Chemie (V.M., M.S., A.S.), Technische Universität München, 85350 Freising-Weihenstephan, Germany; and XL-protein GmbH (A.S.), 85354 Freising, Germany.
Volker
V
Morath
Lehrstuhl für Molekulare Ernährungsmedizin (F.B., A.B., N.R., S.M., M.K.), Else Kröner-Fresenius Center and ZIEL-Research Center for Nutrition and Food Science, and Munich Center for Integrated Protein Science and Lehrstuhl für Biologische Chemie (V.M., M.S., A.S.), Technische Universität München, 85350 Freising-Weihenstephan, Germany; and XL-protein GmbH (A.S.), 85354 Freising, Germany.
Andrea
A
Bast
Lehrstuhl für Molekulare Ernährungsmedizin (F.B., A.B., N.R., S.M., M.K.), Else Kröner-Fresenius Center and ZIEL-Research Center for Nutrition and Food Science, and Munich Center for Integrated Protein Science and Lehrstuhl für Biologische Chemie (V.M., M.S., A.S.), Technische Universität München, 85350 Freising-Weihenstephan, Germany; and XL-protein GmbH (A.S.), 85354 Freising, Germany.
Nadine
N
Rink
Lehrstuhl für Molekulare Ernährungsmedizin (F.B., A.B., N.R., S.M., M.K.), Else Kröner-Fresenius Center and ZIEL-Research Center for Nutrition and Food Science, and Munich Center for Integrated Protein Science and Lehrstuhl für Biologische Chemie (V.M., M.S., A.S.), Technische Universität München, 85350 Freising-Weihenstephan, Germany; and XL-protein GmbH (A.S.), 85354 Freising, Germany.
Martin
M
Schlapschy
Lehrstuhl für Molekulare Ernährungsmedizin (F.B., A.B., N.R., S.M., M.K.), Else Kröner-Fresenius Center and ZIEL-Research Center for Nutrition and Food Science, and Munich Center for Integrated Protein Science and Lehrstuhl für Biologische Chemie (V.M., M.S., A.S.), Technische Universität München, 85350 Freising-Weihenstephan, Germany; and XL-protein GmbH (A.S.), 85354 Freising, Germany.
Sabine
S
Mocek
Lehrstuhl für Molekulare Ernährungsmedizin (F.B., A.B., N.R., S.M., M.K.), Else Kröner-Fresenius Center and ZIEL-Research Center for Nutrition and Food Science, and Munich Center for Integrated Protein Science and Lehrstuhl für Biologische Chemie (V.M., M.S., A.S.), Technische Universität München, 85350 Freising-Weihenstephan, Germany; and XL-protein GmbH (A.S.), 85354 Freising, Germany.
Arne
A
Skerra
Lehrstuhl für Molekulare Ernährungsmedizin (F.B., A.B., N.R., S.M., M.K.), Else Kröner-Fresenius Center and ZIEL-Research Center for Nutrition and Food Science, and Munich Center for Integrated Protein Science and Lehrstuhl für Biologische Chemie (V.M., M.S., A.S.), Technische Universität München, 85350 Freising-Weihenstephan, Germany; and XL-protein GmbH (A.S.), 85354 Freising, Germany.
Martin
M
Klingenspor
D020816:Amino Acid Motifs; D000818:Animals; D001067:Appetite Depressants / Q000008:administration & dosage / Q000009:adverse effects / Q000737:chemistry / Q000627:therapeutic use*; D004305:Dose-Response Relationship, Drug; D002149:Energy Intake / Q000187:drug effects; D004734:Energy Metabolism / Q000187:drug effects*; D005260:Female; D007031:Hypothalamus / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D007279:Injections, Subcutaneous; D020738:Leptin / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000235:genetics / Q000627:therapeutic use; D008099:Liver / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D008297:Male; D008817:Mice, Mutant Strains; D008970:Molecular Weight; D009043:Motor Activity / Q000187:drug effects; D009765:Obesity / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology; D010455:Peptides / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000008:administration & dosage / Q000009:adverse effects / Q000737:chemistry / Q000627:therapeutic use*; D012528:Satiety Response / Q000187:drug effects*; D013047:Specific Pathogen-Free Organisms; D022722:Thermogenesis / Q000187:drug effects; D015431:Weight Loss / Q000187:drug effects
D001067:Appetite Depressants; D020738:Leptin; D010455:Peptides; D011993:Recombinant Fusion Proteins