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Digitalis cardiotoxicity.

Clinical manifestations of digitalis toxicity were clearly described by Withering in 1785. One hundred years later, certain digitalis-induced arrhythmias were inscribed on the smoked drum, and shortly thereafter with the introduction of the electrocardiograph, manifestations of digitalis toxicity as recognized today were recorded in animals and human beings. With popularization of the direct-writing electrocardiograph in the late 1940s and the introduction of digitoxin in recommended doses (that in retrospect appear inappropriately high), the documented prevalence of digitalis toxicity increased rapidly. With increased understanding of the interaction of electrolytes and digitalis and perhaps, and more importantly, the widespread use of digoxin in doses derived largely from its inotropic action and, thus, inappropriately low for the management of many of the arrhythmias, the prevalence of digitalis toxicity began to decline again. In addition, the advent of serum level determinations and the widespread acceptance of the concept of "therapeutic" levels which, although frequently falling short of the desired clinical end point, served to preclude digitalis toxicity. With the decline in the incidence of digitalis toxicity consequent to these factors, some of the digitalis-related arrhythmias that were common are now rarely observed. This report focuses on arrhythmias that are highly specific for digitalis toxicity and on those that now are less commonly encountered. The discussion and classification of the arrhythmias are based on their most probable electrophysiologic mechanism.

Animals↗

Survival and prognosis: investigation of Crataegus extract WS 1442 in congestive heart failure (SPICE)--rationale, study design and study protocol.

UNLABELLED: SPICE is the first, international, randomized, placebo-controlled, double-blind study to investigate the influence of the herbal drug Crataegus Special Extract WS 1442 (hawthorn leaves with flowers) on mortality of patients suffering from congestive heart failure. BACKGROUND: In vitro and experimental animal studies have suggested the following pharmacological modes of action of standardized Crataegus extracts: (1) cAMP-independent positive inotropy; (2) peripheral and coronary vasodilation; (3) protection against ischemia-induced ventricular arrhythmias; (4) antioxidative properties; and (5) anti-inflammatory effects. STUDY DESIGN: In this randomized, placebo-controlled, double-blind, international trial (approximately 120 investigational centers in seven European countries), up to 2300 patients with congestive heart failure, New York Heart Association class II and III and markedly impaired left ventricular function, will be enrolled and treated over a period of 24 months. During this time patients receive either two film-coated tablets of 450 mg of the Special Extract WS 1442 standardized to 84.3 mg of oligomeric procyanidines or matched placebo per day in addition to standard therapy for congestive heart failure, such as diuretics, digoxin or digitoxin, beta-adrenoceptor blockers and angiotensin-converting-enzyme inhibitors. The primary outcome variable is the combined endpoint of cardiac death, non-lethal myocardial infarction, and hospitalization due to progression of heart failure. Secondary outcome variables are total mortality, exercise duration, echocardiographic parameters, quality of life as well as pharmacoeconomic parameters. The first patient was included in October 1998. The trial is expected to be completed at the end of 2002.

Adult↗

Identification and quantification of cardiac glycosides in blood and urine samples by HPLC/MS/MS.

Cardiac glycosides (CG) are of forensic importance because of their toxicity and the fact that very limited methods are available for identification of CG in biological samples. In this study, we have developed an identification and quantification method for digoxin, digitoxin, deslanoside, digoxigenin, and digitoxigenin by high-performance liquid chromatography tandem mass spectrometry (HPLC/MS/MS). CG formed abundant [M + NH4]+ ions and much less abundant [M + H]+ ions as observed with electrospray ionization (ESI) source and ammonium formate buffer. Under mild conditions for collision-induced dissociation (CID), each [M + NH4]+ ion fragmented to produce a dominant daughter ion, which was essential to the sensitive method of selected reaction monitoring (SRM) quantification of CG achieved in this study. SRM was compared with selected ion monitoring (SIM) regarding the effects of sample matrixes on the methodology. SRM produced lower detection limits with biological samples than SIM, while both methods produced equal detection limits with CG standards. On the basis of the HPLC/MS/MS results for CG, we have proposed some generalized points for conducting sensitive SRM measurements, in view of the property of analytes as well as instrumental conditions such as the type of HPLC/MS interface and CID parameters. Analytes of which the molecular ion can produce one abundant daughter ion with high yield under CID conditions may be sensitively measured by SRM. ESI is the most soft ionization source developed so far and can afford formation of the fragile molecular ions that are necessary for sensitive SRM detection. Mild CID conditions such as low collision energy and low pressure of collision gas favor production of an abundant daughter ion that is essential to sensitive SRM detection. This knowledge may provide some guidelines for conducting sensitive SRM measurements of very low concentrations of drugs or toxicants in biological samples.

Animals↗

Synthesis and characterization of digoxin-phospholipid conjugates.

The preparation of immunoreactive derivatives of digoxin for analytical applications is most often carried out by periodate cleavage of the terminal sugar ring (digitoxose) followed by reaction with an enzyme, protein, carrier, or related biological molecules. Here we report an improved and more efficient synthesis which was developed to provide digoxin-phospholipid conjugates useful for liposome immunoassay. The approach used involved the linking of the cleaved digitoxose through a carboxymethyl oxime functionality, which provides much improved yields of readily purified products. The synthetic modification should be applicable to the preparation of analogous phospholipid conjugates involving linkage through a sugar ring (digitoxin, ouabain, and related cardiac glycosides) or to those involving steroids (i.e., 3-digoxigenone) which can be modified to form oxime derivatives remote from key functionalities important for immunorecognition by specific antibody. The characterization of the digoxin-phospholipid conjugates with high-resolution NMR and fast atom bombardment mass spectrophotometry will also be discussed.

Chemical Phenomena↗

Multiresidue screen for cardiotoxins by two-dimensional thin-layer chromatography.

A two-dimensional thin-layer chromatographic method was developed for the qualitative determination of the cardiotoxins oleandrin, gitoxin, digitoxin, gitoxigenin, and grayanotoxins I, II, and III in gastrointestinal contents (stomach, rumen, colon, and cecum contents), feces, and plant material. The cardiotoxins were extracted with dichloromethane. The extract was cleaned up by charcoal and reverse phase solid-phase extraction columns. Analysis was performed by two-dimensional thin-layer chromatography on silica gel plates and visualized by aluminum chloride followed by chloramine T spray. The method detection limits were 0.05 microg/g for oleandrin, 0.1 microg/g for gitoxin, and 0.2 microg/g for the other toxicants in gastrointestinal contents and feces and were 5 times higher in plant material. Four replicate fortifications of bovine rumen contents, bovine feces, and alfalfa at these levels were all well recovered. The diagnostic utility of the method was tested by analyzing samples submitted to the veterinary toxicology laboratory.

Animals↗

Synthesis of 20-hydroxy-, 20-amino-, and 20-nitro-14-hydroxy-21-nor-5 beta,14 beta-pregnane C-3 glycosides and related derivatives: structure-activity relationships of pregnanes that bind to the digitalis receptor.

The preparation of derivatives of 14-hydroxy-21-nor-5 beta,14 beta-pregnane and 5 beta,14 beta-pregnane C-3 alpha-L-rhamnosides and tris-beta-D-digitoxosides is described. These derivatives, possessing a C-17 beta COCH2OH, CH2OH, CO2H, CO2Me, CH2NH2, or CH2NO2 group, bind to the digitalis receptor recognition site of heart muscle as measured in a radioligand binding assay. The 21-norpregnane derivatives consistently show greater binding affinity than the corresponding 20 alpha- and 20 beta-pregnane analogs. The C-20 nitro rhamnoside is comparable to digitoxin in binding affinity. The 17 beta-CH2NO2 group is the most effective replacement for the unsaturated lactone in the binding assay found so far, showing binding affinity comparable to that of the cardiac glycosides.

Binding Sites↗

Cardiac glycoside-like structure and function of 5 beta,14 beta-pregnanes.

5 beta-Reduction and 14 beta-substitution convert the planar progesterone molecule to the cardiac glycoside configuration--A and D rings of the steroid moiety are bent toward the alpha-face relative to the B and C rings. Potency of the 5 beta,14 beta-derivative in a [3H]ouabain binding assay or its ability to inhibit the sodium pump in red blood cells is enhanced by 3 beta-hydroxylation, 20 beta-hydroxylation, and 3 beta-glycosidation. Synthesis of 14,20 beta-dihydroxy-3 beta-(beta-D-glucopyranosyloxy)- 5 beta,14 beta-pregnane from digitoxin is described. The glucoside is 1/20 as potent as ouabain and elicits prominent, sustained, positive inotropy in isolated cardiac muscle.

Animals↗

Cardenolide analogues. 2. 22-Methylenecard-14-enolides.

22-Methylene-3beta-hydroxy-5beta,20(S)-card-14-enolide (11) and 22-methylene-3beta-hydroxy-5beta,20(R)-card-14-enolide (12) were synthesized from digitoxin (1). Attempts to prepare the 14beta-hydroxy-22-methylene analogues were unsuccessful. The 20(R) isomer (12) was found in Na+, K+-ATPase inhibition studies to be twice as active as 14-dehydrogitoxigenin (17). The 20(S) isomer (11) was significantly less active than 17. The hydrolysis of steroid 3beta-tert-butyldimethysilyl ethers was also found to be much more difficult than with nonsteroids.

Adenosine Triphosphatases↗

Three-dimensional quantitative structure-activity relationship analysis of ligand binding to human sequence antidigoxin monoclonal antibodies using comparative molecular field analysis.

The present study indicates that the newly generated human sequence antidigoxin monoclonal antibody (mAb), 1B3, binds digoxin with a different fine specificity binding than our previously obtained human sequence monoclonal antibodies (mAbs) (Ball, W. J.; et al. J. Immunol. 1999, 163, 2291-2298). Uniquely, 1B3 has a higher affinity for digitoxin than digoxin, the immunizing hapten, and a strong requirement for at least one sugar residue linked to the aglycone (-genin). By means of comparative molecular field analysis (CoMFA), the results of competition binding studies for 56 cardiotonic and hormonal steroids were employed to develop three-dimensional quantitative structure-activity relationship (3D-QSAR) models for ligand binding to 1B3 and to three additional human sequence mAbs, as well as the murine antidigoxin mAb 40-50 (Mudgett-Hunter, M.; et al. Mol. Immunol. 1985, 22, 447-488). All five 3D-QSAR models yielded cross-validated q(2) values greater than 0.5, which indicates that they have significant predictive ability. The CoMFA StDevCoeff contour plots, as well as the competition results, indicate that 1B3 binds ligands in a manner distinct from the other four mAbs. The CoMFA contour plots for 40-50 were also compared with the known X-ray crystallographic structure of the 40-50-ouabain complex (Jeffrey, P. D.; et al. J. Mol. Biol. 1995, 248, 344-360) in order to identify correlations between residues in the mAb binding site and specific contour plot regions. These 3D-QSAR models and their respective contour plots should be useful tools to further understand the molecular nature of antibody-antigen interactions and to aid in the redesign or enhancement of therapeutic antibodies.

Amino Acid Sequence↗

Role of P-glycoprotein as a secretory mechanism in quinidine absorption from rat small intestine.

The intestinal transport of quinidine was characterized in rat small intestine, using the Ussing-type chamber under short-circuited conditions. In the short-circuited condition, quinidine transport was predominantly secretory and the transport rate in jejunum was 3.5 times larger in the secretory direction than that in the absorptive direction. The secretion of quinidine was found to be dependent upon its concentration and to be via a carrier-mediated system in both jejunum and ileum. Although the kinetic characteristic of the carrier-mediated secretion of quinidine was very similar in jejunum and ileum, its contribution was much greater in jejunum because of a higher passive diffusion component in ileum. The secretion of quinidine, well-known as an inhibitor of P-glycoprotein, was inhibited significantly and its absorption was enhanced significantly by several substrates of P-glycoprotein including verapamil, diltiazem, and digitoxin in jejunum. These phenomena were also observed by the addition of 2,4-dinitrophenol. Furthermore, the voltage-clamp studies indicated that the inhibition of quinidine secretion occurred in the transcellular pathway. On the other hand, neither tetraethylammonium nor p-aminohippuric acid affected the transport of quinidine. Quinidine was also recognized to inhibit the secretion and to promote the absorption of substrates of P-glycoprotein, chlorpromazine, and verapamil. These results strongly suggest that quinidine is not only an inhibitor but also a substrate of P-glycoprotein and that the P-glycoprotein-mediated secretory flux acts as a barrier to quinidine absorption in the small intestine, especially jejunum.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Spironolactone and warfarin interaction.

Spironolactone has been reported to diminish the hypoprothrombinemic effect of oral anticoagulants in animals and digitoxin blood levels in man by induction of hepatic enzymes. To evaluate this in man, single oral doses of racemic warfarin, 1.5 mg/kg body weight, with and without 200 mg oral spironolactine daily, were administered to 9 normal subjects. Daily venous blood samples were analyzed for 1-stage prothrombin activity, hematocrit, and warfarin content. There was a decrease in hypoprothrombinemic effect (p less than 0.01), an increase in venous hematocrit (p less than 0.001), and no change in warfarin level (p greater than 0.5) during warfarin with spironolactone-induced diuresis over that with warfarin alone. It is concluded that the interaction of warfarin and spironolactone results primarily from the diuresis with consequent concentration of clotting factors and decreased anticoagulant effect.

Adult↗

Regulation of the resting potential of rabbit pulmonary artery myocytes by a low threshold, O2-sensing potassium current.

1. The contributions of specific K+ currents to the resting membrane potential of rabbit isolated, pulmonary artery myocytes, and their modulation by hypoxia, were investigated by use of the whole-cell, patch-clamp technique. 2. In the presence of 10 microM glibenclamide the resting potential (-50 +/- 4 mV, n = 18) was unaffected by 10 microM tetraethylammonium ions, 200 nM charybdotoxin, 200 nM iberiotoxin, 100 microM ouabain or 100 microM digitoxin. The negative potential was therefore maintained without ATP-sensitive (KATP) or large conductance Ca(2+)-sensitive (BKCa) K channels, and without the Na(+)-K+ ATPase. 3. The resting potential, the delayed rectifier current (IK(V)) and the A-like K+ current (IK(A)) were all reduced in a concentration-dependent manner by 4-aminopyridine (4-AP) and by quinine. 4. 4-AP was equally potent at reducing the resting potential and IK(V), 10 mM causing depolarization from -44 mV to -22 mV with accompanying inhibition of IK(V) by 56% and IK(A) by 79%. In marked contrast, the effects of quinine on resting potential were poorly correlated with its effects on both IK(A) and IK(V). At 10 mM, quinine reduced IK(V) and IK(A) by 47% and 38%, respectively, with no change in the resting potential. At 100 microM, both currents were almost abolished while the resting potential was reduced < 50%. Raising the concentration to 1 mM had little further effect on IK(A) or IK(V), but essentially abolished the resting potential. 5. Reduction of the resting potential by quinine was correlated with inhibition of a voltage-gated, low threshold, non-inactivating K+ current, IK(N). Thus, 100 microM quinine reduced both IK(N) and the resting potential by around 50%. 6. The resting membrane potential was the same whether measured after clamping the cell at -80 mV, or immediately after a prolonged period of depolarization at 0 mV, which inactivated IK(A) and IK(V), but not IK(N). 7. When exposed to a hypoxic solution, the O2 tension near the cell fell from 125 +/- 6 to 14 +/- 2 mmHg (n = 20), resulting in a slow depolarization of the myocyte membrane to -35 +/- 3 mV (n = 16). The depolarization occurred without a change in the amplitude of IK(V) or IK(A), but it was accompanied by 60% inhibition of IK(N) at 0 mV. 8. Our findings suggest that the resting potential of rabbit pulmonary artery myocytes depends on IK(N), and that inhibition of IK(N) may mediate the depolarization induced by hypoxia.

Animals↗

Pharmacokinetics of digoxin-specific Fab: effects of decreased renal function and age.

AIMS: To study the influence of age and renal function on digoxin-specific Fab (DS-Fab) pharmacokinetics. METHODS: Sixteen patients (35-91 years) with creatinine clearance ranging from 10.6 to 122.1 ml min(-1) who had been admitted to hospital with severe digoxin or digitoxin self-poisoning were treated with DS-Fab (80 to 800 mg). Plasma DS-Fab concentrations were determined by radioimmunoassay. RESULTS: The mean (s.d.) distribution and elimination half-lives, apparent volume of distribution and total body clearance were 1.1 +/- 0.4 h, 20.2 +/- 7.3 h, 13.1 +/- 5.8 l, and 17.6 +/- 10.8 ml min(-1), respectively. Interindividual variability of DS-Fab total body clearance was linked linearly with the decrease in creatinine clearance or with the increase in age and DS-Fab distribution volume was not dependent on creatinine clearance or age. CONCLUSIONS: The data suggest that DS-Fab should be given to elderly and renal-impaired patients at doses similar to those given to younger or normal renal function patients.

Adult↗

Effects of cardiac glycosides on 24-h ambulatory blood pressure in healthy volunteers and patients with heart failure.

Blood pressure effects of cardiac glycosides in humans have been infrequently reported. Although direct infusion of ouabain or digoxin causes vasoconstriction, indirect effects of cardiac glycosides may have the opposite effect, owing to changes in sympatho-vagal balance. This paper summarises three studies on the effects of cardiac glycosides on circadian blood pressure, utilizing automatic 24-h ambulatory blood pressure measurement (ABPM). In healthy volunteers, 10 days of oral digoxin or digitoxin caused decreases in diastolic blood pressure and heart rate during overnight sleep. No effect was detectable during daytime activities. In patients with heart failure (NYHA stage II), 7 days of oral digoxin also caused a decrease in diastolic blood pressure but only a small increase in systolic pressure during overnight sleep. Again, no effect was detectable during the day. Cardiac glycosides have significant effects on blood pressure, which only appear during overnight sleep, i.e. a phase when sympathetic background activity is lowest. Regular daytime activities may 'overwrite' these effects. Effects of cardiac glycosides on blood pressure may have therapeutic impact, depending on the stage of heart failure and concomitant diseases.

Blood Pressure↗

[Relationship between digoxin-induced cardiac arrhythmias and serum-digoxin levels (author's transl)].

The serum-digoxin level was measured by radio-immunoassay (Immutope) on 245 patients in a coronary care unit. Cardiac arrhythmias were assumed to be digoxin-induced if they disappeared after the drug had been stopped. Patients who had received digitoxin or spironolactone were excluded. The results indicated a normal digoxin range of 1.52 +/- 0.2 ng/ml and a toxic one of 2.78 +/- 0.38 ng/ml. First degree A-V block, atrial ectopic beats and ventricular ectopics with variable coupling intervals were frequently associated with serum-digoxin levels of 1.5-2.5 ng/ml, while higher grade A-V block, atrial tachycardia, bigeminy and atrial tachycardia with block were more frequent with higher serum-digoxin levels (greater than 3.0 ng/ml). Atrial arrhythmias were especially frequent with serum levels above 3.0 ng/ml. This suggests different sensitivities of atrial and ventricular myocardium. Atrial arrhythmias thus in general indicate a higher degree of toxicity at high serum levels, while ventricular ectopic beats occur both with high serum levels and also with increased digoxin sensitivity of the ventricles.

Arrhythmias, Cardiac↗

Effects of saponins on the water solubility of different model compounds.

The influence of saponins on the water solubility of compounds having poor aqueous solubility was investigated with the model compounds digitoxin, rutin and aesculin. The saponins that were tested represent the most abundant structural types. Only slight effects on aqueous solubility were obtained for all the model compounds. These effects include both enhancements and reductions in aqueous solubility depending on the saponin, the concentration of the saponin solution and the model compound. Structure-activity relationships of general significance were not observed. Hence, saponins generally should not be regarded as solubilizers.

Models, Chemical↗

Na+,K(+)-ATPase inhibiting activity of cardiac glycosides from Erysimum cheiranthoides.

We previously reported the isolation of eleven new cardiac glycosides called cheiranthosides I-XI together with two known ones (olitoriside and erysimoside) from the seeds of Erysimum cheiranthoides L. The glycosides were evaluated for their inhibitory activity against Na+,K(+)-ATPase by comparing with typical cardiac glycosides. Two of them, cheiranthoside III and VIII, showed high inhibiting activity which was equivalent to that of digitoxin. Cheiranthoside XI containing a rhamnopyranosyl digitoxopyranosyl moiety and a carboxyl group showed the lowest activity which was similar to that of the inactive aglycone, strophanthidin. Some characteristics in the structure-activity relationship are also discussed.

Animals↗

[Bradycardiac atrial fibrillation after consuming herbal tea].

HISTORY AND CLINICAL FINDINGS: One day after drinking what she thought to be a tea made from borage leaves a 72-year-old woman developed nausea, vomiting and diarrhoea, later also flickering in her eyes and palpitations. She was in a good general state with a blood pressure of 120/75 mm Hg and an irregular heart rate of 52/min. Physical examination was otherwise unremarkable. She had not been on any medication. INVESTIGATIONS: The usual laboratory tests were normal. The electrocardiogram showed atrial fibrillation with a slow ventricular rate with pauses of up to 1.5s. intermittently type I and II 2 degrees AV block, and depressed concave ST segments. The level of digoxin was 3.93 ng/ml, that of digitoxin 133.5 ng/ml. TREATMENT AND COURSE: The patient's symptoms quickly improved under symptomatic treatment. Further questioning suggested that she had probably mistaken foxglove leaves for those of borage when picking them to make a brew. CONCLUSION: If cardiac arrhythmias have occurred after intake of self-picked herbal leaves one should consider digitalis intoxication resulting from misidentification.

Acute Disease↗