Tags

Type your tag names separated by a space and hit enter

Interactions of an Anti-Sickling Drug with Hemoglobin in Red Blood Cells from a Patient with Sickle Cell Anemia.
Bioconjug Chem. 2019 03 20; 30(3):568-571.BC

Abstract

The pathophysiology associated with sickle cell disease (SCD) includes hemolytic anemia, vaso-occlusive events, and ultimately end organ damage set off by the polymerization of deoxygenated hemoglobin S (HbS) into long fibers and sickling of red blood cells (RBCs). One approach toward mitigating HbS polymerization is to pharmacologically stabilize the oxygenated (R) conformation of HbS and thereby reduce sickling frequency and SCD pathology. GBT440 is an α-subunit-specific modifying agent that has recently been reported to increase HbS oxygen binding affinity and consequently delay in vitro polymerization. In addition, animal model studies have demonstrated the potential for GBT440 to be a suitable therapeutic for daily oral dosing in humans. Here, we report an optimized method for detecting GBT440 intermediates in human patient hemolysate using a combination of HPLC and mass spectrometry analysis. First, oxygen dissociation curves (ODCs) analyzed from patient blood showed that oxygen affinity increased in a dose dependent manner. Second, HPLC and integrated mass spectrometric analysis collectively confirmed that GBT440 labeling was specific to the α N-terminus thereby ruling out other potential ligand binding sites. Finally, the results from this optimized analytical approach allowed us to detect a stable α-specific GBT440 adduct in the patient's hemolysate in a dose dependent manner. The results and methods presented in this report could therefore potentially help therapeutic monitoring of GBT440 induced oxygen affinity and reveal critical insight into the biophysical properties of GBT440 Hb complexes.

Links

PMC Free PDF

Authors+Show Affiliations

Strader MB
Laboratory of Biochemistry and Vascular Biology, Center for Biologics Evaluation and Research , Food and Drug Administration (FDA) , Silver Spring , Maryland 20993 , United States.
Liang H
Laboratory of Biochemistry and Vascular Biology, Center for Biologics Evaluation and Research , Food and Drug Administration (FDA) , Silver Spring , Maryland 20993 , United States.
Meng F
Laboratory of Biochemistry and Vascular Biology, Center for Biologics Evaluation and Research , Food and Drug Administration (FDA) , Silver Spring , Maryland 20993 , United States.
Harper J
Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Disease , National Institutes of Health , Bethesda , Maryland 20892-0520 , United States.
Ostrowski DA
Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Disease , National Institutes of Health , Bethesda , Maryland 20892-0520 , United States.
Henry ER
Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Disease , National Institutes of Health , Bethesda , Maryland 20892-0520 , United States.
Shet AS
Sickle Cell Branch, National Heart Lung and Blood Institute , National Institutes of Health , Bethesda , Maryland 20892 , United States.
Eaton WA
Laboratory of Chemical Physics, National Institute of Diabetes and Digestive and Kidney Disease , National Institutes of Health , Bethesda , Maryland 20892-0520 , United States.
Thein SL
Sickle Cell Branch, National Heart Lung and Blood Institute , National Institutes of Health , Bethesda , Maryland 20892 , United States.
Alayash AI
Laboratory of Biochemistry and Vascular Biology, Center for Biologics Evaluation and Research , Food and Drug Administration (FDA) , Silver Spring , Maryland 20993 , United States.

MeSH

Anemia, Sickle CellAntisickling AgentsBenzaldehydesErythrocytesHemoglobin, SickleHumansMolecular Docking SimulationOxygenPyrazinesPyrazoles

Pub Type(s)

Case Reports
Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, P.H.S.

Language

eng

PubMed ID

30794381

Citation

Strader, Michael Brad, et al. "Interactions of an Anti-Sickling Drug With Hemoglobin in Red Blood Cells From a Patient With Sickle Cell Anemia." Bioconjugate Chemistry, vol. 30, no. 3, 2019, pp. 568-571.
Strader MB, Liang H, Meng F, et al. Interactions of an Anti-Sickling Drug with Hemoglobin in Red Blood Cells from a Patient with Sickle Cell Anemia. Bioconjug Chem. 2019;30(3):568-571.
Strader, M. B., Liang, H., Meng, F., Harper, J., Ostrowski, D. A., Henry, E. R., Shet, A. S., Eaton, W. A., Thein, S. L., & Alayash, A. I. (2019). Interactions of an Anti-Sickling Drug with Hemoglobin in Red Blood Cells from a Patient with Sickle Cell Anemia. Bioconjugate Chemistry, 30(3), 568-571. https://doi.org/10.1021/acs.bioconjchem.9b00130
Strader MB, et al. Interactions of an Anti-Sickling Drug With Hemoglobin in Red Blood Cells From a Patient With Sickle Cell Anemia. Bioconjug Chem. 2019 03 20;30(3):568-571. PubMed PMID: 30794381.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Interactions of an Anti-Sickling Drug with Hemoglobin in Red Blood Cells from a Patient with Sickle Cell Anemia. AU - Strader,Michael Brad, AU - Liang,Hongying, AU - Meng,Fantao, AU - Harper,Julia, AU - Ostrowski,David A, AU - Henry,Eric R, AU - Shet,Arun S, AU - Eaton,William A, AU - Thein,Swee L, AU - Alayash,Abdu I, Y1 - 2019/02/28/ PY - 2019/2/23/pubmed PY - 2019/11/30/medline PY - 2019/2/23/entrez SP - 568 EP - 571 JF - Bioconjugate chemistry JO - Bioconjug Chem VL - 30 IS - 3 N2 - The pathophysiology associated with sickle cell disease (SCD) includes hemolytic anemia, vaso-occlusive events, and ultimately end organ damage set off by the polymerization of deoxygenated hemoglobin S (HbS) into long fibers and sickling of red blood cells (RBCs). One approach toward mitigating HbS polymerization is to pharmacologically stabilize the oxygenated (R) conformation of HbS and thereby reduce sickling frequency and SCD pathology. GBT440 is an α-subunit-specific modifying agent that has recently been reported to increase HbS oxygen binding affinity and consequently delay in vitro polymerization. In addition, animal model studies have demonstrated the potential for GBT440 to be a suitable therapeutic for daily oral dosing in humans. Here, we report an optimized method for detecting GBT440 intermediates in human patient hemolysate using a combination of HPLC and mass spectrometry analysis. First, oxygen dissociation curves (ODCs) analyzed from patient blood showed that oxygen affinity increased in a dose dependent manner. Second, HPLC and integrated mass spectrometric analysis collectively confirmed that GBT440 labeling was specific to the α N-terminus thereby ruling out other potential ligand binding sites. Finally, the results from this optimized analytical approach allowed us to detect a stable α-specific GBT440 adduct in the patient's hemolysate in a dose dependent manner. The results and methods presented in this report could therefore potentially help therapeutic monitoring of GBT440 induced oxygen affinity and reveal critical insight into the biophysical properties of GBT440 Hb complexes. SN - 1520-4812 UR - https://www.unboundmedicine.com/medline/citation/30794381/Interactions_of_an_Anti_Sickling_Drug_with_Hemoglobin_in_Red_Blood_Cells_from_a_Patient_with_Sickle_Cell_Anemia_ DB - PRIME DP - Unbound Medicine ER -