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Related Concept Videos

Phase II Reactions: Miscellaneous Conjugation Reactions01:19

Phase II Reactions: Miscellaneous Conjugation Reactions

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Phase II biotransformations are detoxification mechanisms that conjugate xenobiotics with endogenous substances, neutralizing their toxicity.
A key example involves the conjugation of cyanide ions, which impair cellular respiration and alter hemoglobin into non-oxygen-carrying cyanmethemoglobin. To neutralize this threat, a sulfur atom from thiosulphate is transferred to the cyanide ion, catalyzed by the enzyme rhodanese, resulting in an inactive compound called thiocyanate. The production of...
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Drug Metabolism: Phase II Reactions01:14

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Phase II reactions are essential for the detoxification and elimination of drugs from the body. These reactions involve the conjugation of parent drugs or their phase I metabolites with endogenous molecules, resulting in more hydrophilic drug conjugates. The primary conjugation reactions in this phase are sulfation and glucuronidation. Both sulfation and glucuronidation typically produce biologically inactive metabolites. However, in some cases involving prodrugs, active metabolites may be...
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Phase II Reactions: Sulfation and Conjugation with α-Amino Acids01:19

Phase II Reactions: Sulfation and Conjugation with α-Amino Acids

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Sulfation and α-amino acid conjugation are two critical biotransformation reactions in drug metabolism. Sulfation, a phase II biotransformation reaction, involves adding a polar sulfate group to a drug, enhancing its water solubility and promoting excretion. This process can either co-occur with or occur independently of glucuronidation. Nonmicrosomal sulfotransferase enzymes catalyze the process. The reaction involves 3'-phosphoadenosine-5'-phosphosulfate or PAPS coenzyme...
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Phase II Reactions: Glutathione Conjugation and Mercapturic Acid Formation01:22

Phase II Reactions: Glutathione Conjugation and Mercapturic Acid Formation

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Glutathione, a tripeptide made up of glutamate, cysteine, and glycine, is a critical player in the detoxification of drugs and xenobiotics via a process known as glutathione conjugation or mercapturic acid formation. This phase II biotransformation reaction involves the covalent binding of glutathione to a drug or its metabolite, enhancing the compound's water solubility and enabling its excretion.
Several distinctive characteristics distinguish glutathione conjugation from other phase II...
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Phase II Conjugation Reactions: Overview01:14

Phase II Conjugation Reactions: Overview

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Conjugation, a key component of phase II biotransformation reactions, is a vital process in drug detoxification. It involves transferring endogenous substances like glucuronic acid, sulfate, and glycine to drugs or their metabolites formed in phase I reactions. These conjugation reactions, often catalyzed by specific enzymes, transform potentially harmful metabolites into inactive, water-soluble forms easily excreted in urine or bile. By enhancing polarity and eliminating pharmacological...
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Phase II Reactions: Glucuronidation01:24

Phase II Reactions: Glucuronidation

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Glucuronidation, a pivotal phase II biotransformation process, involves the coupling of glucuronic acid to a drug or xenobiotic. Given its widespread occurrence and critical role in drug metabolism, it's considered the most crucial phase II reaction. It enhances the water solubility of substances, aiding their expulsion from the body. The driving force behind these reactions is a group of enzymes known as UDP-glucuronosyltransferases (UGTs). UGTs facilitate the transfer of a glucuronic acid...
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Synthesis of Protein Bioconjugates via Cysteine-maleimide Chemistry
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Bridged Cysteine Conjugations.

Matthew Bird1, Joao Nunes2, Mark Frigerio2

  • 1Abzena Ltd, Cambridge, UK. matthew.bird@abzena.com.

Methods in Molecular Biology (Clifton, N.J.)
|October 24, 2019
PubMed
Summary
This summary is machine-generated.

This study introduces a novel cysteine rebridging method for creating homogeneous antibody-drug conjugates (ADCs). This technique achieves high drug-to-antibody ratios (DAR) on native antibodies without genetic or enzymatic modification.

Keywords:
Cysteine rebridgingHomogeneous drug loadingInterchain disulfideMichael additionThioBridge® conjugation

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Area of Science:

  • Bioconjugation Chemistry
  • Antibody Engineering
  • Drug Development

Background:

  • Homogeneous antibody-drug conjugates (ADCs) require site-selective payload attachment.
  • Current methods often involve antibody engineering or enzymatic modification.
  • Limited methods exist for homogeneous ADCs using endogenous amino acid conjugation.

Purpose of the Study:

  • To develop a robust method for producing ADCs with homogeneous drug loading.
  • To achieve site-selective conjugation at native interchain disulfides.
  • To enable homogeneous ADC production without antibody pre-modification.

Main Methods:

  • Utilized a cysteine rebridging approach for antibody conjugation.
  • Employed a cascade of addition-elimination reactions under mild aqueous conditions.
  • Applied the method to wild-type antibodies without recombinant or enzymatic engineering.

Main Results:

  • Achieved highly homogeneous drug-to-antibody ratios (DAR) at native interchain disulfides.
  • Demonstrated high conversion rates (70-95%) to conserved DAR ADCs.
  • Reported overall process yields greater than 70%.

Conclusions:

  • The ThioBridge® method provides a robust platform for homogeneous ADC preparation.
  • This approach simplifies ADC manufacturing by using wild-type antibodies.
  • The method offers efficient and site-selective conjugation for improved ADC homogeneity.