Trimesoyltris(3,5-dibromosalicylate): specificity of reactions of a trifunctional acylating agent with hemoglobin

Kluger, Ronald; Song, Yonghong; Wodzinska, Jolanta; Head, Charlotte; Fujita, T.S.; Jones, Richard T.

doi:10.1021/ja00050a005

Cited by 42 publications

(65 citation statements)

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“…The cross-linked hemoglobin was stored at 4°C under carbon monoxide. The purity of the cross-linked hemoglobin was found to be Ͼ99% cross-linked between the ␤-chains by C 4 reverse-phase HPLC according to previously described methodology (24). The absence of monomeric ␤-chains was demonstrated by matrix-assisted laser desorption/ionization time-of-flight mass spectroscopy using an Applied Systems Voyager DE STR equipped with a 337-nm nitrogen laser (results not shown).…”

Section: Human Hemoglobin Cross-linked With Trimesoylmentioning

confidence: 96%

“…The solution was subjected to gel chromatography (Sephadex G-25 column, 5 ϫ 25 cm, equilibrated with 0.1 M MOPS buffer, pH 8.0). Hemoglobin was then deoxygenated with nitrogen at 37°C for 2 h before the deoxygenated hemoglobin was cross-linked at the two ␤-Lys82 residues by reaction with a twofold excess of trimesoyl tris(3Ј,5Ј-dibromosalicylate) (16 mol, 16 mg) for 16 h at 37°C (24). The mixture was bubbled with oxygen to stop the cross-linking reaction, and the tetrameric hemoglobin was separated from unreacted trimesoyl tris(3Ј,5Ј-dibromosalicylate) by size-exclusion chromatography using a Sephadex G-25 column (2.5 ϫ 25 cm) equilibrated with 0.1 M phosphate buffer, pH 7.4.…”

Section: Human Hemoglobin Cross-linked With Trimesoylmentioning

confidence: 99%

“…We have undertaken a thorough comparison of native human hemoglobin and a ␤-chain [trimesyl-(Lys82)␤-(Lys82)␤] cross-linked human hemoglobin (24) to understand the pharmacokinetic properties of synthetic, acellular hemoglobins in rats in vivo. Because it is recognized that the formation of a complex between hemoglobin and haptoglobin results in liver cell entry and clearance, binding of the cross-linked human hemoglobin to rat haptoglobin should also result in hepatocellular entry and degradation in rats.…”

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confidence: 99%

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Binding of acellular, native and cross-linked human hemoglobins to haptoglobin: enhanced distribution and clearance in the rat

Ship

Toprak

Lai

et al. 2005

American Journal of Physiology-Gastrointestinal and Liver Physi

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Section: Human Hemoglobin Cross-linked With Trimesoylmentioning

confidence: 96%

Section: Human Hemoglobin Cross-linked With Trimesoylmentioning

confidence: 99%

mentioning

confidence: 99%

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Binding of acellular, native and cross-linked human hemoglobins to haptoglobin: enhanced distribution and clearance in the rat

Ship

Toprak

Lai

et al. 2005

American Journal of Physiology-Gastrointestinal and Liver Physi

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“…␤-Chain [trimesoyl-(Lys82)␤-(Lys82)␤] cross-linked human hemoglobin was prepared according to Kluger et al (1992) and described previously (Ship et al, 2005). Carbon-monoxide-liganded hemoglobin (2 ml, 4 mM) was exposed to bright white light and bubbled with oxygen (BOC Gases, Mississauga, ON, Canada) for 2 h at 0°C to exchange the carbon monoxide with oxygen.…”

Section: Materials Radiolabeled [1-mentioning

confidence: 99%

Role of Haptoglobin on the Uptake of Native and β-Chain [Trimesoyl-(Lys82)β-(Lys82)β] Cross-Linked Human Hemoglobins in Isolated Perfused Rat Livers

Chow

Liu²,

Ship³

et al. 2008

Drug Metab Dispos

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ABSTRACT:The role of haptoglobin in liver cell entry of acellular native hemoglobin, and cross-linked human hemoglobin, a potentially useful oxygen-carrier alternative in transfusion medicine, was examined in the recirculating, perfused rat liver preparation. Doses of tritiated native human or ␤-chain [trimesoyl-(Lys82)␤-(Lys82)␤] crosslinked human hemoglobin were preincubated with haptoglobincontaining rat plasma or Krebs Henseleit bicarbonate buffer for 30 min and used for perfusion. Concentrations (dpm/ml) in reservoir, before and after separation of the hemoglobins and metabolites by gel filtration fast protein liquid chromatography column chromatography, were similar, showing mostly the presence of intact hemoglobin. Each hemoglobin species underwent a rapid distribution phase, followed by a protracted elimination phase. The radioactivity in bile at 3 h consisted of low molecular weight metabolites, and cumulative excretion was slightly higher when rat plasma was present: for native hemoglobin, 7.1 ؎ 1.6% versus 9.2 ؎ 2.1% dose; for cross-linked hemoglobin, 5.0 ؎ 1.7% versus 7.2 ؎ 0.8% dose. Data fit to a two-compartment model and physiologically based model revealed a significantly faster influx clearance (CL influx ) over the metabolic intrinsic clearance (CL int, met ). The ratios of CL influx /CL int, met were 125 and 535 for native hemoglobin in the absence and presence of rat haptoglobin, respectively, according to compartmental analyses; the ratios were 25 and 53, respectively, according to physiological modeling. The corresponding ratios for cross-linked hemoglobin in the absence and presence of rat haptoglobin were 55 and 81, respectively, and 24 and 70 for compartmental and physiological modeling. Although haptoglobin enhanced the hepatic internalization of the hemoglobins, the impact on the net clearance was lessened since degradation was the rate-limiting step.Hemoglobin is a tetrameric heme protein that is found within red blood cells. This protein is composed of a pair of ␣␤ dimers to form its tetrameric structure (Greer et al., 1981;Voet and Voet, 1995;Chang, 1999). In the tetrameric format, hemoglobin plays an important role in the binding to oxygen and the supply of oxygenated blood to tissues and organs [for review, see Winslow (1992)]. During intravascular hemolysis, hemoglobin is released from the red blood cells and is distributed into the plasma where free hemoglobin (HB) is rapidly bound to haptoglobin (Hp), a specific plasma binding protein of high affinity (K d Ϸ 1 ϫ 10 Ϫ15 M) toward the alpha chain on the hemoglobin (Putnam, 1976;McCormick and Atassi, 1990;Lim et al., 1998). The complex is then taken up by an unknown receptor on hepatocytes and by the CD163 hemoglobin scavenger receptor on macrophages that are present in the spleen, thymus, Kupffer cells, and bone marrow (Graversen et al., 2002;Polfliet et al., 2006).In cases of transfusion of acellular hemoglobin or severe hemolysis, high levels of hemoglobin are present and will easily saturate the haptoglobin present in plasma (...

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“…The retained functional group permits the addition of nucleophilic materials, which could be delivered as a hemoglobin conjugate in circulation (25,26).…”

Section: Reaction Patterns Of Methyl Acyl Phosphatesmentioning

confidence: 99%

1994 Syntex Award Lecture: Anionic Electrophiles, Protein Modification, and Artificial Blood

Kluger¹

1994

Can. J. Chem.

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Monomethyl esters of acyl phosphates (mixed anhydrides of carboxylic and phosphoric acids) are being developed as site-directed acylating agents for amino groups in proteins. In an illustrative application, these anionic materials are shown to bind to a positively charged region of hemoglobin where they convert amino groups to amides. Bifunctional acyl methyl phosphates cross-link hemoglobin to give a variety of products, some of which have the oxygen-binding properties anticipated for materials that can serve as an alternative to red cells in transfusions. Higher yields of desired products result from the use of a trifunctional analogue. Kinetic patterns of the reactions of a series of alkyl amines and methyl aroyl phosphates indicate that the transition state for formation of the amide involves almost complete development of positive charge on nitrogen.

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Trimesoyltris(3,5-dibromosalicylate): specificity of reactions of a trifunctional acylating agent with hemoglobin

Cited by 42 publications

References 0 publications

Binding of acellular, native and cross-linked human hemoglobins to haptoglobin: enhanced distribution and clearance in the rat

Binding of acellular, native and cross-linked human hemoglobins to haptoglobin: enhanced distribution and clearance in the rat

Role of Haptoglobin on the Uptake of Native and β-Chain [Trimesoyl-(Lys82)β-(Lys82)β] Cross-Linked Human Hemoglobins in Isolated Perfused Rat Livers

1994 Syntex Award Lecture: Anionic Electrophiles, Protein Modification, and Artificial Blood

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