Albumin-binding domain extends half-life of glucagon-like peptide-1

Abstract Background Long-acting glucagon-like peptide-1 receptor agonists (GLP-1 RAs) are widely used for treatment of type 2 diabetes (T2DM) in the United States, the European Union, and Japan. In our previous work, we designed and characterized a novel GLP-1 RA, SKL-18287. This RA consists of only natural L-amino acids, and is believed to exist in an oligomer form in systemic circulation. This unique feature may allow high biological stability and a long-lasting glucose lowering effect in T2DM treatment. In the present study, we investigated the pharmacokinetic properties of SKL-18287 in rats, monkeys, and mini-pigs. Tissue distributions of radioactivity were also studied in rats after subcutaneous administration of [3H]-SKL-18287. Methods Plasma concentrations of SKL-18287 were measured by LC-MS/MS after intravenous and subcutaneous administration of SKL-18287 in rats, monkeys, and mini-pigs. Pharmacokinetic parameters were then calculated and compared among these animal species. Tissue concentrations of radioactivity were determined by liquid scintillation counting following sample combustion, after subcutaneous administration of [3H]-SKL-18287 to rats. Results SKL-18287 showed an extended half-life of over 5 h, with good subcutaneous bioavailability, in all animal species. Prediction of the pharmacokinetic profiles of SKL-18287 in humans using an animal scale-up approach revealed an SKL-18287 half-life of 14.8 h. The radioactivity concentration in the pancreas, a target tissue of GLP-1RA, was relatively higher than in other tissues, until 12 h after [3H]-SKL-18287 administration. Conclusion SKL-18287 might be sufficient to maintain an effective concentration for a once-daily treatment for T2DM, and is a unique GLP-1 RA with a pancreas-selective feature.

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A GLP-1 receptor agonist conjugated to an albumin-binding domain for extended half-life

Biopolymers ◽

10.1002/bip.22474 ◽

2014 ◽

Vol 102 (3) ◽

pp. 252-259 ◽

Cited By ~ 15

Author(s):

Joel Lindgren ◽

Essam Refai ◽

Sergei V. Zaitsev ◽

Lars Abrahmsén ◽

Per-Olof Berggren ◽

...

Keyword(s):

Half Life ◽

Receptor Agonist ◽

Binding Domain ◽

Albumin Binding

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Glucagon-like peptide 1 undergoes differential tissue-specific metabolism in the anesthetized pig

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.1996.271.3.e458 ◽

1996 ◽

Vol 271 (3) ◽

pp. E458-E464 ◽

Cited By ~ 55

Author(s):

C. F. Deacon ◽

L. Pridal ◽

L. Klarskov ◽

M. Olesen ◽

J. J. Holst

Keyword(s):

Immunosorbent Assay ◽

Half Life ◽

Biologically Active ◽

Enzyme Linked Immunosorbent Assay ◽

Metabolic Clearance ◽

Tissue Specific ◽

Glucagon Like Peptide ◽

Glucagon Like Peptide 1 ◽

Plasma Half Life ◽

Differential Tissue

Glucagon-like peptide 1 (GLP-1) metabolism was studied in halothane-anesthetized pigs (n = 7) using processing-independent (PI) and COOH-terminal (C) radioimmunoassays (RIA) and an enzyme-linked immunosorbent assay (ELISA) specific for biologically active GLP-1. Renal extraction of endogenous GLP-1 was detected by PI-RIA (33.1 +/- 13.3%) and C-RIA (16.0 +/- 6.3%) and by all assays during GLP-1 infusion (ELISA, 69.4 +/- 6.3%; PI-RIA, 32.6 +/- 7.3%; C-RIA, 43.7 +/- 3.4%), indicating substantial fragmentation. Hepatic and pulmonary degradation were undetectable under basal conditions, but exogenous GLP-1 elimination by the liver (43.6 +/- 8.9%) and lungs (10.1 +/- 3.2%) was measured by ELISA, suggesting primarily NH2-terminal degradation. Endogenous GLP-1 extraction by the hindleg was only detected by C-RIA (16.0 +/- 6.3%). During GLP-1 infusion, greater hindleg extraction was measured by ELISA (38.5 +/- 6.8%) and C-RIA (33.0 +/- 6.4%) than by PI-RIA (11.4 +/- 3.2%), indicating limited degradation at each terminus or more substantial COOH-terminal degradation. A shorter (P < 0.01) plasma half-life was revealed by ELISA (1.5 +/- 0.4 min) than by PI-RIA (4.5 +/- 0.6 min) or C-RIA (4.1 +/- 0.5 min). Metabolic clearance rates measured by PI-RIA (20.0 +/- 3.8 ml.min-1.kg-1) and C-RIA (15.5 +/- 1.6 ml.min-1.kg-1) were shorter (P < 0.01) than that measured by ELISA (106.8 +/- 14.7 ml.min-1.kg-1). Tissue-specific differential metabolism of GLP-1 occurs, and NH2-terminal degradation, rendering GLP-1 inactive, is particularly important in its clearance.

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Fusion of an albumin-binding domain extends the half-life of immunotoxins

International Journal of Pharmaceutics ◽

10.1016/j.ijpharm.2016.07.046 ◽

2016 ◽

Vol 511 (1) ◽

pp. 538-549 ◽

Cited By ~ 20

Author(s):

Rui Guo ◽

Wenjun Guo ◽

Li Cao ◽

Hui Liu ◽

Jieyu Liu ◽

...

Keyword(s):

Half Life ◽

Binding Domain ◽

Albumin Binding

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Faculty Opinions recommendation of Extending half-life by indirect targeting of the neonatal Fc receptor (FcRn) using a minimal albumin binding domain.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.718913136.793527659 ◽

2017 ◽

Author(s):

Chava Kimchi-Sarfaty

Keyword(s):

Half Life ◽

Fc Receptor ◽

Binding Domain ◽

Albumin Binding ◽

Neonatal Fc Receptor

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Peptide Half-Life Extension: Divalent, Small-Molecule Albumin Interactions Direct the Systemic Properties of Glucagon-Like Peptide 1 (GLP-1) Analogues

Journal of Medicinal Chemistry ◽

10.1021/acs.jmedchem.7b00787 ◽

2017 ◽

Vol 60 (17) ◽

pp. 7434-7446 ◽

Cited By ~ 17

Author(s):

Esben M. Bech ◽

Manuel C. Martos-Maldonado ◽

Pernille Wismann ◽

Kasper K. Sørensen ◽

Søren Blok van Witteloostuijn ◽

...

Keyword(s):

Small Molecule ◽

Half Life ◽

Life Extension ◽

Glucagon Like Peptide ◽

Glucagon Like Peptide 1

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Extension of human GCSF serum half-life by the fusion of albumin binding domain

Scientific Reports ◽

10.1038/s41598-021-04560-6 ◽

2022 ◽

Vol 12 (1) ◽

Author(s):

Fatemeh Yadavar Nikravesh ◽

Samira Shirkhani ◽

Elham Bayat ◽

Yeganeh Talebkhan ◽

Esmat Mirabzadeh ◽

...

Keyword(s):

Half Life ◽

Animal Studies ◽

Cytoplasmic Inclusion ◽

Binding Domain ◽

Size Exclusion ◽

Albumin Binding ◽

Exclusion Chromatography ◽

Aggregation Formation

AbstractGranulocyte colony stimulating factor (GCSF) can decrease mortality of patients undergo chemotherapy through increasing neutrophil counts. Many strategies have been developed to improve its blood circulating time. Albumin binding domain (ABD) was genetically fused to N-terminal end of GCSF encoding sequence and expressed as cytoplasmic inclusion bodies within Escherichia coli. Biological activity of ABD-GCSF protein was assessed by proliferation assay on NFS-60 cells. Physicochemical properties were analyzed through size exclusion chromatography, circular dichroism, intrinsic fluorescence spectroscopy and dynamic light scattering. Pharmacodynamics and pharmacokinetic properties were also investigated in a neutropenic rat model. CD and IFS spectra revealed that ABD fusion to GCSF did not significantly affect the secondary and tertiary structures of the molecule. DLS and SEC results indicated the absence of aggregation formation. EC50 value of the ABD-GCSF in proliferation of NFS-60 cells was 75.76 pg/ml after 72 h in comparison with control GCSF molecules (Filgrastim: 73.1 pg/ml and PEG-Filgrastim: 44.6 pg/ml). Animal studies of ABD-GCSF represented improved serum half-life (9.3 ± 0.7 h) and consequently reduced renal clearance (16.1 ± 1.4 ml/h.kg) in comparison with Filgrastim (1.7 ± 0.1 h). Enhanced neutrophils count following administration of ABD-GCSF was comparable with Filgrastim and weaker than PEG-Filgrastim treated rats. In vitro and in vivo results suggested the ABD fusion as a potential approach for improving GCSF properties.

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HER2-Specific Pseudomonas Exotoxin A PE25 Based Fusions: Influence of Targeting Domain on Target Binding, Toxicity, and In Vivo Biodistribution

Pharmaceutics ◽

10.3390/pharmaceutics12040391 ◽

2020 ◽

Vol 12 (4) ◽

pp. 391

Author(s):

Haozhong Ding ◽

Mohamed Altai ◽

Wen Yin ◽

Sarah Lindbo ◽

Hao Liu ◽

...

Keyword(s):

Half Life ◽

Binding Domain ◽

Protein Domain ◽

Specific Cell ◽

Albumin Binding ◽

Exotoxin A ◽

Pseudomonas Exotoxin ◽

Pseudomonas Exotoxin A ◽

Number Of Patients ◽

Plasma Half Life

The human epidermal growth factor receptor 2 (HER2) is a clinically validated target for cancer therapy, and targeted therapies are often used in regimens for patients with a high HER2 expression level. Despite the success of current drugs, a number of patients succumb to their disease, which motivates development of novel drugs with other modes of action. We have previously shown that an albumin binding domain-derived affinity protein with specific affinity for HER2, ADAPT6, can be used to deliver the highly cytotoxic protein domain PE25, a derivative of Pseudomonas exotoxin A, to HER2 overexpressing malignant cells, leading to potent and specific cell killing. In this study we expanded the investigation for an optimal targeting domain and constructed two fusion toxins where a HER2-binding affibody molecule, ZHER2:2891, or the dual-HER2-binding hybrid ZHER2:2891-ADAPT6 were used for cancer cell targeting. We found that both targeting domains conferred strong binding to HER2; both to the purified extracellular domain and to the HER2 overexpressing cell line SKOV3. This resulted in fusion toxins with high cytotoxic potency toward cell lines with high expression levels of HER2, with EC50 values between 10 and 100 pM. For extension of the plasma half-life, an albumin binding domain was also included. Intravenous injection of the fusion toxins into mice showed a profound influence of the targeting domain on biodistribution. Compared to previous results, with ADAPT6 as targeting domain, ZHER2:2891 gave rise to further extension of the plasma half-life and also shifted the clearance route of the fusion toxin from the liver to the kidneys. Collectively, the results show that the targeting domain has a major impact on uptake of PE25-based fusion toxins in different organs. The results also show that PE25-based fusion toxins with high affinity to HER2 do not necessarily increase the cytotoxicity beyond a certain point in affinity. In conclusion, ZHER2:2891 has the most favorable characteristics as targeting domain for PE25.

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