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Gold compounds in rheumatic diseases--1.

Gold complexes diminish synovitis and improve therapeutic outcome in rheumatoid arthritis, psoriatic arthritis, some forms of juvenile chronic arthritis, and palindromic rheumatism. The decision to treat a patient with a gold compound should not be undertaken lightly. It should be understood by the patient that the commitment to therapy is potentially long-term, that major benefit is not assured, and that there is, approximately, a 50% probability of an adverse reaction which may terminate treatment. Nevertheless, most adverse effects are mild and transient and the probability of good control, even remission, in an otherwise threatening disease, is sufficiently high to justify a therapeutic trial in the majority of patients. Those patients whose rheumatoid arthritis appears to respond to gold may be encouraged to continue long-term therapy with maintenance doses, in view of the evidence of sustained efficacy in most patients and of the declining risk of adverse reactions after the first 12 months.

Adult↗

Effects of chrisotherapeutic gold compounds on prostaglandin E2 production.

The mechanism of action of anti-rheumatic gold compounds on 12-O-tetradecanoylphorbol 13-acetate (TPA)-induced prostaglandin E(2) (PGE(2)) production in rat peritoneal macrophages were examined. Auranofin (AF) at 3-10 muM inhibited TPA-induced PGE(2) production in a concentration-dependent manner. In the pharmacological experiments, prostaglandin G/H synthase (PGHS)-2-dependent PGE(2) production was inhibited by 10 muM of AF. The enzyme activities of both PGHS-1 and PGHS-2 were not affected by the 10 muM AF. Other gold compounds, aurothioglucose (ATG) and aurothiomalate (ATM) did not inhibit PGE2 production at 10 muM. AF decreased the PGHS-2 protein content, but had no effect on the PGHS-1 protein content. AF at 3-10 muM decreased the PGHS-2 messenger RNA (mRNA) level by RT-PCR determination. Then, the effect of AF on nuclear factor kappa B (NF-kappaB), one of the transcription factors known to regulate transcription of a group of proinflammatory proteins, was determined. AF at 1-10 muM inhibited nuclear translocation of NF-kappaB in a concentration-dependent manner. ATG and ATM at 10 muM did not inhibit NF-kappaB nuclear translocation, but with 20 h preincubation, ATG and ATM inhibited PGE(2) production and NF-kappaB nuclear translocation. AF, ATG, and ATM did not affect the binding of NF-kappaB to its specific DNA. These observations may suggest that the effects of gold compounds on the inhibition of NF-kappaB nuclear translocation plays one of the major role in its anti-inflammatory effects in rat peritoneal macrophages.

Animals↗

Anti-rheumatic gold compounds as sublethal modulators of monocytic LPS-induced cytokine secretion.

The objective of this study was to quantify the ability of sublethal concentrations of several gold compounds to differentially modulate the monocytic secretion of key cytokines that are important in the etiology of rheumatic diseases. Human THP1 monocytic cells were exposed to the anti-rheumatic drugs auranofin (AF), gold sodium thiomalate (GSTM) or HAuCl4 (Au(III)) for 24-72 h. Succinate dehydrogenase (SDH) activity of the monocytes was used to determine sublethal concentrations. Monocytes were then exposed to sublethal concentrations of gold compounds for 72 h, and the activator lipopolysaccharide (LPS) was added (or not) to cultures for the last 6h. The secretion of IL6, IL8, IL10, and TNFalpha were measured in cell supernatants using ELISA. Cytokine secretion was compared among concentrations and gold compounds. SDH experiments established a sublethal concentration range of 0-75 microM for GSTM and Au(III) and 0-0.5 microM for AF. In cytokine experiments, none of the compounds alone activated secretion of any of the cytokines, but all differentially (50-440%, p<0.05) increased LPS-induced secretion of IL6 and IL8 over TNFalpha and IL10. In conclusion, sublethal concentrations of AF, GSTM, and Au(III) all may differentially modulate activation of monocytic cells, and this differential modulation may be important in the mechanisms of action of these compounds.

Antirheumatic Agents↗

Ploidity and cell cycle progression during treatment with gold chloride, auranofin and sodium aurothiomalate. Studies on a human epithelial cell line and its sub-strains made resistant to the antiproliferative effects of these gold compounds.

The possible influence of gold(III) chloride and the two gold(I)-containing anti-arthritic drugs, auranofin and sodium aurothiomalate, on cellular ploidity and cell cycle progression was investigated on cultured human epithelial cells. Four different cell lines were used: the parent line (HE) and three sub-strains which previously had acquired resistance to the antiproliferative effects of either 350 mumol gold chloride/l culture medium (HEAu350), 2 mumol auranofin/l (HEAF) or 300 mumol sodium aurothiomalate/l (HEMyo). DNA-histograms were obtained by flow cytometry examinations during a 9-days' exposure to either of these gold-containing compounds and concentrations. The HE, HEAF and HEMyo cells had similar ploidities, close to tetraploid. The HEAu350 cells had altered ploidity to distinct tetraploid. The distribution of the resistant cells with the cell cycle phases was not different from that of untreated HE cells. The HE cells, when treated with auranofin or sodium aurothiomalate, accumulated in the G2-phase of the cell cycle. In addition, a new cedecoploid peak appeared. No such changes were observed on gold chloride exposure or in HE controls grown without drug supplement. The effects of auranofin and sodium aurothiomalate on cell cycle progression of the HE cells possibly indicate a tendency to polyploidity, and furthermore that inhibition of cellular mitosis is one mechanism of the antiproliferative effect common to the two drugs.

Auranofin↗

Gold compounds in rheumatoid arthritis. Report of a symposium.

A vast accumulation of clinical experience and controlled studies justifies the use of gold compounds in selected cases of rheumatoid arthritis. The striking therapeutical efficacy observed in some patients has stimulated active research to clarify the pharmacokinetics and mechanisms of action of gold compounds. Based on this knowledge, the possibility of selecting gold-responsive patients may be enhanced, and the frequency of adverse reactions decreased. Liberal indications and inexperience in administration and control have given gold therapy mixed reputation. A symposium in Umeå, Sweden, summarized present knowledge and current concepts among Scandinavian rheumatologists on the rational use of gold compounds.

Arthritis, Rheumatoid↗

[Mechanism of action of gold compounds].

The mechanisms of action of gold salts are still poorly understood. While Auranofin (AF) has a direct anti-inflammatory action in animal models, Myochrysin (MC) has not. Both gold compounds inhibit various enzymes. Among these, lysosomal enzymes are of particular interest. Gold is known to accumulate in the lysosomes, and gold salts modulate certain macrophage functions in the immune system. The role of organic gold salt ligands containing, like D-penicillamine, a thiol group is still unclear. Positive data characterizing responders versus nonresponders are not yet available. As in other clinically heterogeneous diseases, medical action has mainly been based on clinical empiricism which, although resting on false premises, has helped to find a comparatively efficacious treatment. In fact, the proposals for further studies to shed light on the mode of action presented here may equally be ill conceived. However, these studies would at least expand our understanding of rheumatoid arthritis and thus be beneficial in the long run.

Animals↗

[Intracellular gold content of circulating blood cells using various gold compounds].

Evidence on the action mechanisms of gold salts in the treatment of rheumatoid arthritis is still inconclusive. The intracellular localization of the place of action is likely. Therefore not only the serum gold levels but also the intracellular concentration of gold are of special interest. We measured the gold concentration in the serum and in the blood cells after in vitro application of aurothiomalate (Tauredon), gold keratinate (Auro-Detoxin) and triethylphosphine-gold (Ridaura) and in blood samples of patients undergoing these gold salts treatments. Cell-bound concentrations were found to vary extensively as a function of the gold compound used. While no or very little gold was present intracellularly after administration of the 2 parenteral drugs, up to 40% of the circulating gold was found to bind to the cells after administration of the triethylphosphine compound for gastro-intestinal absorption. The red cell concentration was more or less the same as that in the extracellular compartment. Gold apparently accumulated in the white cells, because the cell-bound concentration relative to unit volume was up to 20 times higher than the plasma level. The method used did not offer any information on the actual binding site of gold in white cells, i.e. cytoplasm versus nucleus versus cell membrane.

Anti-Inflammatory Agents↗

Comparative pharmacokinetics of parenteral and oral gold compounds.

The pharmacokinetics of intramuscular and oral gold compounds differ widely. Aurothioglucose and gold sodium thiomalate absorption is complete; 25% of auranofin (AF) is absorbed. Blood gold concentrations with conventional parenteral treatment generally peak between 600-800 microgram/dl the day of injection and decline gradually to about 300 micrograms/dl 7 days later. Levels range between 30 and 100 micrograms/dl, using 2-9 mg/d AF, and show little variation. A smaller percentage of gold is found in the cellular fraction of blood with I.M. than with oral gold. The blood half-life is approximately 6 days with gold sodium thiomalate, and 21 days with AF. Forty percent of the administered dose of injectable gold is excreted; depending upon dosage, 75-100% of oral gold is recovered in excreta, which is a combination of unabsorbed and excreted gold. Nearly 70% of parenteral gold is excreted in the urine and 30% in feces, while only 5% of AF is in urine and 95% in feces. The amount of gold retained following intravenous 195Au-labelled gold sodium thiomalate is 43% at 60 days and 25% at 250 days, but only 15% with oral radiolabeled AF 10 days after ingestion. Synovial fluid gold levels are much higher with parenteral than with oral gold but the blood-to-synovial fluid ratio is similar. Skin gold concentrations rise steadily with injectable but not oral treatment, but hair and nail accumulation is insignificant. Corneal, lens, and skin chrysiasis may develop with parenteral therapy, but has not been recognized with AF.

Administration, Oral↗

Effect of gold compounds on phorbol myristic acetate (PMA) activated superoxide (O2-) production by mouse peritoneal macrophages.

A number of gold compounds, nonsteroidal antiinflammatory drugs (NSAID) and protease inhibitors have been examined for their ability to inhibit the respiratory burst of peritoneal macrophages. A potent (IC50 = 5 microM or less) dose dependent inhibitory activity was seen with a number of triethylphosphine gold I ( TEPG ) compounds. Nonphosphine gold compounds were less active. The ligands of TEPG were inactive. Several NSAID stimulated the respiratory burst, whereas 2 protease inhibitors tested were inhibitory. The structure activity relationship of TEPG is also presented.

Animals↗

Helicobacter pylori seroprevalence in patients with rheumatoid arthritis: effect of nonsteroidal anti-inflammatory drugs and gold compounds.

PURPOSE: To investigate the relationship between Helicobacter pylori infection and nonsteroidal anti-inflammatory drug (NSAID) intolerance and the effect of gold use on the seroprevalence of H. pylori. PATIENTS AND METHODS: We examined the frequency of discontinuation of NSAIDs in 132 unselected patients with rheumatoid arthritis attending an outpatient subspecialty clinic, and the effect of gold compound use on the seroprevalence (by IgG enzyme-linked immunosorbent assay) of H. pylori infection in this population. Logistic and multivariate regression analysis was performed adjusting for age, gender, ethnic origin, history of ulcer, and duration of rheumatoid arthritis. RESULTS: Fifty-four patients had a positive serology for H. pylori (41%). Twenty-seven of the seropositive patients (50%), versus 45 of the seronegative patients (57.7%), had to discontinue NSAIDs (aspirin and/or nonaspirin) at least once since their diagnosis of rheumatoid arthritis because of gastrointestinal side effects (odds ratio [OR], 0.93; 95% confidence interval [CI], 0.63 to 1.38). Forty-one of the seropositive patients (76%) had received gold compounds as compared with 62 of the seronegative patients (79.5%) (OR: 0.96; 95% CI: 0.61 to 1.50). CONCLUSION: We did not find any relationship between H. pylori seropositivity and NSAID intolerance in patients with rheumatoid arthritis. In addition, our results do not demonstrate a reduction in H. pylori seroprevalence in rheumatoid arthritis patients treated with gold compounds.

Aged↗

Effect of gold compounds on NADPH oxidase system of human neutrophils.

The inhibitory effects of gold compounds on the NADPH oxidase system of human polymorphonuclear leukocytes (PMNs) has been investigated. Auranofin (0.5-4.0 micrograms Au/ml) suppressed the rate of superoxide anion generation as well as the total yield in cells stimulated with phorbol myristate acetate and f-Met-Leu-Phe. This implies that drug action may be occurring at the level of protein kinase C or steps subsequent to this in the signal transduction sequence. Sodium aurothiomalate (1-100 micrograms Au/ml) lacked such activity. Neither gold compound altered the ability of the granule-rich fraction of PMNs to produce oxy radicals whether this fraction was obtained from drug-treated cells or was treated after its isolation. Therefore, in order for auranofin to exhibit its inhibitory effects on the NADPH oxidase system, an intact cell membrane is necessary.

Auranofin↗

Inhibition of neovascularization in vivo by gold compounds.

As mononuclear cell infiltration and growth of pannus critically depend on synovial neovascularization in rheumatoid arthritis (RA), inhibition of the synovial blood vessels would have the potential to reduce rheumatoid inflammation. In this investigation, we studied the effect of gold sodium thiomalate (GST) and auranofin (AUR) on neovascularization in vivo by using a micropocket technique. Both GST and AUR suppressed rabbit corneal neovascularization in a dose-dependent fashion. Significant inhibition was observed by 3 mg/kg GST and 1 mg/kg AUR injected intravenously every other day. These injections maintained serum gold concentrations at the level of 2-5 micrograms/ml and less than 2 micrograms/ml in GST- and AUR-injected rabbits, respectively. These are concentrations attained in the serum or synovium of rheumatoid patients treated by gold compounds. Similar inhibition was observed by both intramuscular administration of GST and oral administration of AUR. In contrast, no inhibition was observed when non-steroidal anti-inflammatory drugs (NSAIDs; 20 mg/kg acetylsalicylic acid, 10 mg/kg ibuprofen and 10 mg/kg indomethacin) were injected intravenously on a daily basis. These results suggested that gold compounds have an antiangiogenic effect in vivo. The inhibition of neovascularization by gold compounds suggested that they may suppress rheumatoid synovitis by reducing the number of small blood vessels required for mononuclear cell infiltration and synovial tissue proliferation.

Animals↗

The comparative evaluation of the physical and chemical properties of gold compounds.

A wide range of modern physical and chemical techniques can be used to evaluate the properties of gold compounds. These allow the identification of the design features which control their pharmacological activity: the oxidation state (formal charge) on the gold atom, number and types of atoms bonded to gold (coordinated ligands), the stereochemical arrangement of ligands around gold (coordination geometry), and specific targeting groups in the ligands. The polymeric, hydrophilic, gold thiolate compounds are contrasted with the monomeric, lipophilic triethylphosphine gold compound auranofin (AF). The dynamic ligand exchange reactions of the 2 classes of drugs differ markedly; however other reactions are possible, and may lead to some common intermediates in vivo. This conclusion is based on preliminary results using nuclear magnetic resonance spectroscopy, a technique which allows us to monitor reactions in intact cells.

Aurothioglucose↗

Effect of gold compounds on the activity of adenylyl cyclase in human lymphocyte membranes.

OBJECTIVE: To determine the effects of aurothioglucose, aurothiomalate, and auranofin on basal and forskolin-activated adenylyl cyclase activity in human total lymphocyte membranes, and in membranes of T and B lymphocyte subsets. METHODS: Membranes were isolated from human total lymphocytes and T and B cell subsets. The effects of gold compounds on basal and forskolin-stimulated activity of adenylyl cyclase were measured by radioassay. RESULTS: The gold compounds inhibited adenylyl cyclase activity. This inhibitory effect required the presence of both the sulfhydryl ligands and aurous cation. CONCLUSION: Regulation of lymphocyte adenylyl cyclase by gold compounds represents a potential mode of action of these drugs in rheumatic diseases.

Adenylyl Cyclases↗

Evaluation of the in vivo antitumor activity and in vitro cytotoxic properties of auranofin, a coordinated gold compound, in murine tumor models.

The coordinated gold compound, 2,3,4,6-tetra-O-acetyl-1-thio-beta-D-glucopyranosato-S-triethyl phosphine gold (auranofin; Ridaura), was evaluated for antitumor activity in a variety of mouse tumor models. Of the 15 tumor models evaluated, auranofin was found to be active only against i.p. P388 leukemia. A number of dose schedules was used to measure activity against P388 with optimal activity observed at 12 mg/kg given daily, i.p., on Days 1 to 5. Auranofin was active against i.p. P388 leukemia only when administered i.p.; the drug was completely inactive when administered i.v., s.c., or p.o. on Days 1 to 5. Evaluation of the effects of auranofin in vitro demonstrated that survival curves for B16 melanoma cells as measured by the clongenic and dye exclusion assays were exponential and monophasic; cell cycle distribution was not altered, and auranofin displayed no preferential cytotoxicity to logarithmic or plateau growth phase cell populations; auranofin inhibited DNA, RNA, and protein synthesis at cytotoxic concentrations but showed no selective effect; the cytotoxic activity and cellular association of gold from auranofin were dose, time, and temperature dependent; and binding of auranofin gold to serum proteins markedly decreased cellular uptake of gold and cytotoxicity of auranofin in vitro.

Animals↗

Inhibition of human endothelial cell proliferation by gold compounds.

Neovascularization has a role in the propagation of rheumatoid synovitis because the spread of mononuclear cell infiltration and the growth of pannus are dependent on the growth of new blood vessels. Growth of such vessels requires local endothelial cell (EC) proliferation. Inhibition of synovial EC proliferation, therefore, would have the potential to diminish rheumatoid inflammation. We have, therefore, studied the effects of gold sodium thiomalate (GST), auranofin, and gold chloride on the proliferation of human umbilical vein EC. GST suppressed both basal and EC growth factor-induced tritiated thymidine incorporation into EC in a dose-dependent fashion. Inhibition was observed with concentrations as low as 1 microgram/ml GST, 5 micrograms/ml gold chloride, and 0.1 microgram/ml auranofin, levels attainable in blood and synovium of patients. These results suggest that gold compounds have an antiangiogenic effect. The low concentrations inhibiting EC proliferation suggest that gold compounds may suppress rheumatoid synovitis by reducing the number of small blood vessels available for mononuclear cell infiltration and synovial tissue proliferation.

Arthritis, Rheumatoid↗

Low-dose gold compounds inhibit fibroblast proliferation and do not affect interleukin-1 secretion by macrophages.

We examined the effect of low concentrations of gold compounds on the proliferation of human fibroblasts. Gold sodium thiomalate (GST) inhibited both basal and interleukin-1-induced tritiated thymidine incorporation into fibroblasts in a dose- and time-dependent manner. Significant inhibition was observed at the level of 5 micrograms/ml GST, and greater than 50% inhibition was attained at 10 micrograms/ml. These concentrations are attainable in the serum of treated patients. Similar inhibition was observed when less than 1 micrograms/ml auranofin, which is also within a serum-attainable range, was added. Low concentrations of GST (0-10 micrograms/ml) did not affect interleukin-1 secretion from lipopolysaccharide-stimulated human mononuclear phagocytes (M phi) when assessed by both human fibroblast and C3H/HeJ mouse thymocyte proliferation assays. When M phi precultured for 48 hours with GST (0-10 micrograms/ml) were added to the fibroblast culture in the presence or absence of lipopolysaccharide, there was no significant inhibition of M phi-induced DNA synthesis of fibroblasts. In contrast, when fibroblasts were precultured with GST (0-10 micrograms/ml) for 48 hours and freshly separated M phi were added, significant inhibition was observed in M phi-induced fibroblast proliferation at 5 micrograms/ml. These results suggest that low concentrations of GST directly cause a reduction of fibroblast proliferation, but do not affect the capability of M phi for induction of fibroblast proliferation. Therefore, gold compounds may play a role in the inhibition of the growth of rheumatoid pannus by direct inhibition of fibroblast proliferation.

Auranofin↗

Anti-oxidant effects of gold compounds.

The anti-oxidant efficacy, in vitro, of the gold compounds auranofin (AF) and gold sodium thiomalate (GST) was examined by studying their effects on the generation of reactive oxygen species (ROS) using zymosan-stimulated polymorphonuclear leukocytes (PMNs) and a cell-free, xanthine-xanthine oxidase system. The oxygen species investigated were the superoxide radical anion (O2-), hydrogen peroxide (H2O2) and the hydroxyl radical (OH.). AF had an inhibitory effect on ROS production by PMNs. In particular, OH. generation was significantly suppressed in a dose-dependent fashion. AF did not inhibit ROS production in the cell-free system. GST produced only a small degree of inhibition at higher concentrations. These findings suggest that AF may play an important role in the inhibition of respiratory bursts and the generation of inflammatory reaction products. Since the products of the respiratory burst, especially potent oxidants such as OH. and H2O2, are thought to be important inflammatory mediators, it is postulated that the blockade of toxic ROS generation by AF affects rheumatoid as well as dermatological inflammation and tissue damage.

Antioxidants↗