Drug-protein conjugates—XV: A study of the disposition of D-penicillamine in the rat and its relationship to immunogenicity

doi:10.1016/0006-2952(88)90148-7

Biochemical Pharmacology

Volume 37, Issue 4, 15 February 1988, Pages 737-742

https://doi.org/10.1016/0006-2952(88)90148-7 Get rights and content

Abstract

The disposition of [¹⁴C]d-penicillamine (PA) was investigated in vitro and in vivo with male Wistar rats. Irreversible binding of [¹⁴C]PA to isolated rat plasma proteins in vitro reached a maximum of 20.6% of total radioactivity at 6 hr. Irreversibly bound [¹⁴C]PA could be dissociated with dithiothreitol, demonstrating that conjugation was via disulphide linkage. Three hours after i.v. administration of [¹⁴C]PA (27 μmol/kg) to rats 100% of plasma radioactivity was irreversibly bound, representing approximately 3.5% of the dose. Further studies on the disposition of PA-plasma protein conjugates snowed that dissociation occurred readily in vivo: the plasma half-life of the conjugate was approximately 3 hr. Free [¹⁴C]PA was the major urinary metabolite after administration of both free and conjugated drug. These studies show that the disposition of PA is similar to that reported for the structurally related sulphydryl drug captopril (CP). Free PA (340 μmol/kg and 3.4 mmol/kg) administered i.p. and i.m. daily for 4 days at one monthly intervals, and also PA-KLH conjugate (100 μg/rat) administered by single i.p. injection at monthly intervals with and without Freund's complete adjuvant, failed to induce PA-specific IgG or IgM antibody responses detectable by ELISA. In contrast, CP (270μmol/kg), administered by the same protocol as free PA, induced a CP-specific IgG antibody response after the third series of monthly injections. These data suggest that the difference in immunogenicity between PA and CP arises from a difference in the intrinsic immunogenicity of the haptens, rather than from their disposition.

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Free-radical degradation of high-molecular-weight hyaluronan induced by ascorbate plus cupric ions. Testing of bucillamine and its SA981-metabolite as antioxidants
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Citation Excerpt :
The typical daily oral dose of d-pen administered to RA patients is 0.5–2.0 g. The serum half-life of a single oral dose of the drug is 1–3 h [11]. l-Cysteine (Fig. 1e), a naturally occurring amino acid, contributes to the intracellular thiol pool and is a precursor of intracellular l-glutathione biosynthesis [6,12,13].
High-molecular-weight hyaluronan (HA) samples were exposed to free-radical chain-degradation reactions induced by ascorbate in the presence of Cu(II) ions – the so-called Weissberger's oxidative system. The concentrations of both reactants [ascorbate, Cu(II)] were comparable to those that may occur during an early stage of the acute phase of joint inflammation. The time-dependent changes of the viscosity of the HA solution in the absence of the substance tested were monitored by rotational viscometry. However, when the anti- or pro-oxidative effects of the antioxidants/drugs were investigated, their dose-dependency was also examined. Additionally, the anti-oxidative activities of these substances were screened by the well-established ABTS and DPPH decolorization assays. The actions of the disease-modifying anti-rheumatic drugs, namely bucillamine and d-penicillamine, were compared to those of l-cysteine and of SA981, the oxidized metabolite of bucillamine.
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T cell responses to D-penicillamine in drug-induced myasthenia gravis: Recognition of modified DR1:peptide complexes
1999, Journal of Neuroimmunology
The anti-rheumatoid drug d-penicillamine (d-pen) has a reactive sulfhydryl group capable of modifying self antigens, and can provoke typical autoantibody-mediated myasthenia gravis (MG), especially in DR1⁺ individuals. We have selected T cell clones from one such patient that were highly specific for d-pen but not its l-isomer or d-cysteine. Moreover, they were restricted to HLA-DR1, had a Th1 phenotype and used TCR Vα4.1, Vβ6.1. They responded well to blood mononuclear cells prepulsed with d-pen either in the absence of serum or after chloroquine treatment, but not to autologous d-pen-pulsed B cell lines. Thus, d-pen may directly couple to distinctive peptides resident in surface DR1 molecules on circulating macrophages or dendritic cells.
Chapter 19. Chemical Basis for Immune Mediated Idiosyncratic Drug Hypersensitivity
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Idiosyncratic drug reactions are unexpected and unpredictable. While they affect a relatively small number of patients, their adverse reactions can be fatal and are, therefore, of considerable concern. A large number of reports of the reactions are based on the clinical observations of symptoms rather than a definitive demonstration of drug- or metabolite- specific antibodies or immune lymphocytes in the patient. This chapter discusses those classes of compounds for which there is significant evidence of an immune-mediated etiology. Definitive structure/toxicity relationships are often hard to discern, but specific examples illustrate some underlying principles of idiosyncratic adverse drug reactions. Methodology for an adverse drug reaction to qualify as immune-mediated, hapten-reactive antibodies or cells must be demonstrated in sensitized test subjects using one or more of the assays—haptenization of proteins and cells, aliphatic thiols, beta lactams, halothanes, quinone precursors, hydantoins, anilines, aryl sulfur compounds, and aryl acetic acids.
<sup>1</sup>H-Nuclear magnetic resonance study of the oxidation/reduction chemistry of penicillamine in intact human erythrocytes
1990, BBA - Molecular Cell Research
The oxidation/reduction chemistry of penicillamine in human erythrocytes was characterized directly in intact erythrocytes by ¹H-NMR spectroscopy. Spectra were measured by the Car-Purcell-Meiboom-Gill pulse sequence to selectively eliminate interefering resonances from hemoglobin and membrane protons and from the intracellular water. Glucose-free penicillamine-containing erythrocytes were subjected to oxidative stress by titration with t-butyl hydroperoxide. The t-butyl hydroperoxide rapidly crosses the erythrocyte membrane and reacts with gluthathione (GSH) and penicillamine (PSH), with the PSH being oxidized to penicillamine-glutathione mixed disulfide (PSSG) as indicated by the appearance of characteristic resonances in the high resolution ¹H-NMR spectrum. Extracellular PSH is oxidized to penicillamine disulfide (PSSP) when t-butyl hydroperoxide is added to the cells and thereafter in amounts dependent on the oxygenation state of the sample. Following addition of glucose, both the oxidized glutathione and the PSSG are rapidly reduced at comparable rates. The results of additional experiments using erythrocyte lysate and of kinetic experiments on solutions containing PSSG and/or GSH, NADPH and glutathione reductase suggest that the predominant mechanism for reduction of PSSG is by a thiol-disulfide exchange reaction with GSH to form PSH and GSSG, which in turn undergoes enzyme-catalyzed reduction by NADPH.
Effect of inhibition of glutathione synthesis on the immunogenicity of captopril and captoprilprotein conjugates in the mouse
1990, Biochemical Pharmacology
The effect of glutathione (GSH) depletion on the immunogenicity of captopril (CP) and d-penicillamine (PA) was investigated in C57 mouse. Depletion of GSH was by dl-buthionine sulfoximine (BSO), a potent and specific inhibitor of GSH synthesis, injected on the days of immunization of the drug/drug-protein conjugates. Chronic BSO pretreatment for 5 consecutive days, before and including the 4-day immunization period, caused tissue GSH depletion (liver 60%, kidney 71%, lung 14%, spleen 14% and whole blood 36%) in C57 mouse. After chronic administration of CP (270 μmol/kg, i.p. or i.m.) or CP-HSA (200 μg, i.p. or i.m.), IgG antibody response to CP-derived antigen was detected by an enzyme-linked immunosorbent assay (ELISA). The IgG antibody recognised CP-OVA but not OVA, and was inhibited by CP in other protein conjugated form, thus confirming its specificity to CP. Depletion of GSH by BSO caused an increase in the CP-specific IgG antibody titre when CP/CP-HSA was immunized through i.p. (with or without Freund's Complete Adjuvant) but not i.m. administration. In contrast, chronic administration of PA (270 $μmol kg$ , i.p.) or PA-HSA (200μg, i.p.), with or without BSO pre-treatment, did not lead to detectable PA-specific IgG antibody. The results again illustrate the difference in the intrinsic immunogenicity between CP and PA, in that depletion of GSH increased the humoral (Blymphocyte) anti-CP responses but not anti-PA responses under the experimental conditions. These findings suggest that GSH status, apart from its effects on the disposition of CP/CP-protein conjugates, should be considered as an important determinant of both the immunological and toxicological response to CP.
Effect of inhibition of glutathione synthesis on the metabolism and protein conjugation of [<sup>14</sup>C]captopril in the rat
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^∗: Present address: Department of Pharmacology, Chinese University of Hong Kong.

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Drug-protein conjugates—XV: A study of the disposition of D-penicillamine in the rat and its relationship to immunogenicity

Abstract

Biochem. Pharmac.

Biochem. Pharmac.

Chem. Biol. Interact.

Biochem. Pharmac.

Biochem. Pharmac.

Biochem. Pharmac.

Lancet

Distamine Data Sheet