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Microbial inhibition by pharmaceutical antibiotics in different soils--dose-response relations determined with the iron(III) reduction test.

Soil contamination from pharmaceuticals is an emerging problem, though quantitative data on their microbial effects are lacking. Thus, nine pharmaceutical antibiotics were tested for their effects on the microbial iron(III) reduction in six different topsoils. Complete dose-response curves were obtained and best-fit by sigmoidal Logit, Weibull, Box-Cox Logit, and Box-Cox Weibull equations (r2 0.73-1.00). The derived effective doses (ED [micromol/kg soil]) for the different antibiotics increased in the order (average ED50 in parentheses) chlortetracycline (53) < sulfadimethoxine (58) < oxytetracycline (170) < sulfadiazine (190) < sulfadimidine (270) = tetracycline (270) < sulfapyridine (430), though no effect was found for sulfanilamide and fenbendazole at doses up to 5,800 and 3,300 micromol/kg, respectively. Due to a strong soil adsorption, especially of the tetracyclines, the corresponding effective concentrations in the soil solution (EC50), derived from sorption experiments, were considerably smaller and ranged from 0.004 micromol/L (chlortetracycline) to 120 micromol/L (sulfapyridine). The effects of the antibiotics were governed by soil sorptive properties, especially the concentration of soil organic matter. The microbial inhibition was influenced indirectly by the soil pH, which affects the ionization status of the amphoteric antibiotics.

Adsorption↗

Haemolytic potential of three chemotherapeutic agents and aspirin in glucose-6-phosphate dehydrogenase deficiency.

The potential haemolytic effect of three chemotherapeutic drugs and aspirin was tested in vitro by gluthathione stability tests. Blood was collected from the local population of Basra, Iraq where previous studies had found a high frequency of glucose-6-phosphate dehydrogenase (G6PD) deficiency. Primaquine, chloramphenicol and sulfanilamide caused significant concentration-dependent reductions of glutathione levels in G6PD-deficient red cells when compared to normal red cells. Acetylsalicylic acid had no effect on glutathione level. The G6PD-deficient erythrocytes behaved as previously reported, probably due to similar patterns in the distribution of its variants. Studies on each local variant are warranted and new drugs should be tested for haemolytic potential prior to their introduction in areas where the deficiency is common.

Anti-Bacterial Agents↗

Urine flow dependence of renal clearance and interrelation of renal reabsorption and physicochemical properties of drugs.

A physiologically based model is presented for the analysis of the reabsorption kinetics of drugs in the renal tubules. The two reabsorption parameters, AR(I) X Pe and sigma, expressing the permeability of a drug through lipoidal membranes and the reflection coefficient, respectively, are obtained on the basis of this model using the results of clearance experiments in rats. These two reabsorption parameters are compared among four drugs, sulfanilamide, sulfamethizole, theophylline, and ethanol. Urine flow dependence of the percentage of reabsorbed drugs is reasonably expressed by the model. Moreover, the percentage of reabsorbed drugs which are highly reflected at the membranes is affected by the change of the glomerular filtration rate. In contrast, the percentage of reabsorbed drugs, such as ethanol, having a small reflection coefficient is not sensitive to the glomerular filtration rate. A good linear relationship is obtained between AR(I) X Pe and the n-octanol/water partition coefficient. The reflection coefficient is related to the molecular weight of the compound. The reflection coefficient of ethanol is small because of its low molecular weight, whereas the reflection coefficient is almost unity when the molecular weight is higher than 170. Accordingly, this physiological model approach can be acceptable to the reabsorption kinetics of drugs in the renal tubules.

Absorption↗

[Effect of nontoxic inhibitors on the nitrosation of drugs under conditions simulating the human stomach].

The influence of sulphamic acid, sulfanilamide, p-aminosalicylic acid and ascorbic acid on the nitrosation behaviour of Analgin (active agent: noramidopyrinmethane sulfonate) was investigated in the pH range between 1.2 and 6.0 under simulated conditions of the human stomach. Using a colorimetric measuring method, the three last-mentioned compounds proved suitable as nitrosation inhibitors; preference should be given to ascorbic acid because of its better inhibitory capacity in the very weakly acidic pH range. Sulphamic acid is inadequate because it stimulates nitrosation in this pH range. The distinct inhibitory action of ascorbic acid was verified under simulated conditions of the human stomach in further 9 drugs undergoing nitrosation at different rates, which contain the active agents: aminophenazone, clomipramine, imipramine, desipramine, ampicilline, oxacilline, phenoxymethylpenicilline, ethambutole or piperazine. Despite the use of high doses of ascorbic acid, no complete inhibition of nitrosation could be achieved in the majority of drugs investigated.

Aminopyrine↗

Simple colorimetric method for determination of pyridoxine hydrochloride (vitamin B6) in pharmaceuticals.

A simple colorimetric method is described for the determination of pyridoxine hydrochloride (vitamin B6). The method is based on the measurement of an orange species formed when pyridoxine hydrochloride is treated with diazotized dapsone and sulfanilamide in a mixture of trichloroacetic acid and sulfuric acid at room temperature, followed by treatment with an aqueous solution of sodium carbonate. Compounds such as thiamine hydrochloride, cyanocobalamin, and common excipients such as starch and talc which are present in various formulations with pyridoxine hydrochloride do not interfere in the reaction. Statistical validation showed that the method was highly precise and accurate. Results agree well with those obtained by other methods reported in the literature.

Colorimetry↗

[Bullous toxidermatosis and HIV infection in hospital environment in Lome (Togo)].

A transversal study allowed to analyse the different features of severe bullous toxicodermatosis, and its relations with HIV infection. Since March 1992 to December 1995, 15 patients were hospitalized in Lome Teaching Hospital for severe bullous toxicodermatosis: 16 cases of Lyell's syndrome, 8 cases of Stevens-Johnson's syndrome and 1 case of ectodermosis. The principal drug which induced these diseases were mostly dominated by the sulfanilamide (n = 7). Nine (9) of these subjects were infected by the HIV (four of whose patients had Lyell's syndrome). Its noted five cases of death, all in the patients with HIV infection.

Adult↗

Inhibition of drug metabolism by hydroxylated metabolites: cross-inhibition and specificity.

Inhibition of drug metabolism was studied in adult male Sprague-Dawley rats. A hydroxylated metabolite of phenylbutazone (oxyphenbutazone) inhibited the elimination of phenytoin, which is metabolized by oxidative pathways. The biotransformation of a relatively polar and only slightly plasma protein-bound drug, antipyrine, was subject to product inhibition by a hydroxylated metabolite, 4-hydroxyantipyrine. Neither oxyphenbutazone nor 4-hydroxyantipyrine measurably affected the elimination kinetics or metabolic fate of a drug (sulfanilamide) that is not metabolized by oxidative pathways.

Animals↗

Facile separation of sulfonamides from their degradates by liquid--liquid extraction.

Regulation of acidity for protonation of the free N4-amine can provide for the selective liquid--liquid extraction isolation of a sulfonamide from its degradation products. This principle is applied for the stability-indicating determination of sulfacetamide in the presence of sulfanilamide, sulfaquinoxaline in feed, and sulfabromomethazine in dosage forms. In solution, sulfabromomethazine can exhibit photodecomposition to sulfamethazine. The mean relative errors of the these methods and the precision, represented by relative standard deviations, are each typically less than 2%.

Animal Feed↗

Preparation of spherical matrixes of prolonged-release drugs from liquid suspension.

A simple and less expensive method for the preparation of spherical matrixes of prolonged-release drugs was developed from liquid suspension. The drugs dispersed in distilled water were agglomerated with a small amount of organic solution or melt of matrix material. The process, called a "solution" or "melting" method, depended on the state of matrix material. In the present study, sulfamethoxazole and sulfanilamide were agglomerated with white beeswax and ethylcellulose. Furthermore, the parameters affecting the average size and release behavior of the resultant matrixes were investigated. Increasing the amount of matrix material used yielded large matrixes and prolonged-release action. The drugs rendered hydrophobic by surface treatment produced larger matrixes that released drug more slowly than untreated drugs. The sizes of the matrixes decreased and became uniform with increasing agitation speed. The sulfamethoxazole matrixes with wax proved to be enteric coated since the release rate was fast in the alkaline test solution (pH 7.5) but slow in the acidic one (pH 1.2).

Delayed-Action Preparations↗

Quantitative investigation on renal handling of drugs in rabbits, dogs, and humans.

A renal clearance method based on a computer analysis after administration of a single dose of drug was developed for measuring the renal handling of several drugs in rabbits, dogs, and humans. Secretion and reabsorption of sulfamethizole, sulfanilamide, cephalexin, and ampicillin in the nephron were analyzed quantitatively using the plasma concentration and the urinary excretion rate of the drugs. The validity of the proposed model was demonstrated. It appears that tubular secretion of sulfamethizole, cephalexin, and ampicillin depend on the active transport system which is described by Michaelis-Menten kinetics; the tubular reabsorption of these drugs is expressed by first-order kinetics. The maximum velocity of renal secretion per unit weight of these drugs was much higher in rabbits than in dogs or humans. Reabsorption showed similar values in dogs and humans. These findings suggest that an analysis of the renal handling of drugs in dogs might provide useful information when considering the appropriate therapeutic dose in humans.

Adult↗

Spectrophotometric determination of serum nitrite and nitrate by copper-cadmium alloy.

A macro and micro assay for the spectrophotometric determination of serum nitrite and nitrate was developed. Nitrite/nitrate in biological samples can be estimated in a single step by this method. The principle of the assay is the reduction of nitrate by copper-cadmium alloy, followed by color development with Griess reagent (sulfanilamide and N-naphthylethylenediamine) in acidic medium. This assay is sensitive to 1 microM nitrate and is suitable for different biological fluids, including sera with a high lipid concentration. The copper-cadmium alloy used in the present method is easy to prepare and can completely reduce nitrate to nitrite in an hour. The present method provides a simple, cost-effective assay for the estimation of stable oxidation products of nitric oxide in biological samples.

Alloys↗

Molecular cloning of EcoRII endonuclease and methylase genes.

The genes for restriction-modification system EcoRII have been cloned from plasmid N3 DNA using RSF2124 as a vector plasmid. The hybrid plasmids designated pFK321 and pFK322 contained a 5.8 megadaltons EcoRI--fragment derived from N3 DNA including the genes for restriction-modification system EcoRII and a gene for resistance to sulfanilamide.

Bacteriophage lambda↗

Inhibition of intracellular Ca2+ signalling, cytotoxicity and antitumor activity of the herbicide oryzalin and its analogues.

PURPOSE: Studies were conducted on oryzalin (3,5-dinitro-N,N-di(n-propyl)sulfanilamide), a widely used dinitroaniline sulfonamide herbicide, which was identified from plant extracts as an inhibitor of mitogen- and growth factor-mediated intracellular free Ca2+ ([Ca2+]i) signalling in mammalian cells. METHODS AND RESULTS: Oryzalin inhibited vasopressin, bradykinin and platelet-derived growth factor [Ca2+]i signalling in Swiss 3T3 fibroblasts with IC50 values of 14, 16 and 18 microM, respectively. 45Ca2+ uptake into nonmitochondrial stores of saponin-permeabilized Swiss 3T3 cells was inhibited by oryzalin with an IC50 of 34 microM. Oryzalin inhibited colony formation of HT-29 colon carcinoma cells with an IC50 of 8 microM and inhibited the growth of a number of other cancer cell lines and primary human tumors in vitro with IC50 values in the range 3 to 22 microM. A number of oryzalin analogues were studied and an association was found between the ability to inhibit [Ca2+]i signalling and inhibition of the growth of HT-29 human colon cancer cells (P = 0.001) and of CCRF-CEM human leukemia cells (P = 0.016). Oryzalin at doses up to 600 mg/kg administered orally or subcutaneously daily to mice for 3 to 10 days beginning a day after tumor inoculation inhibited the growth of murine B16 melanoma by 63% but showed no appreciable activity when administered subcutaneously or intraperitoneally to mice beginning a number of days after tumor inoculation against a variety of human tumor xenografts. The peak plasma concentration of oryzalin following repeated subcutaneous administration of oryzalin at 600 mg/kg per day to mice was 37 microM and of its major metabolite N-depropyl oryzalin was 53 microM. CONCLUSION: It is unlikely that the absence of significant antitumor activity of oryzalin is a result of the inability to achieve adequate plasma concentrations.

3T3 Cells↗

Sulfaquinoxaline inhibition of vitamin K epoxide and quinone reductase.

Sulfaquinoxaline (N1-(2-quinoxalinyl)sulfanilamide) has been shown to be a potent (Ki = 1 microM) freely reversible inhibitor of the dithiothreitol-dependent reduction of both vitamin K epoxide and vitamin K quinone by rat liver microsomes in vitro. This observation provides an explanation for the hemorrhagic syndrome occasionally seen in poultry on medicated feed and the efficacy of sulfaquinoxaline in anticoagulant based rodenticides. Sulfaquinoxaline inhibition resembled inhibition by coumarin anticoagulants (e.g., warfarin) and hydroxynaphthoquinones (e.g., lapachol). Inhibition was observed in assays using microsomes from control strain rats, but the enzyme was resistant to sulfaquinoxaline in microsomes from warfarin-resistant rats. Steady-state kinetics inhibition patterns were nearly competitive versus dithiothreitol and nearly uncompetitive versus vitamin K epoxide as is observed for warfarin and lapachol. These results suggest that this inhibitor binds to the oxidized form of vitamin K epoxide reductase in the same way as suggested for the coumarins and hydroxyquinones. Of 10 other sulfa drugs tested, none were inhibitors, and of fragments and related compounds tested, only 2-aminoquinoxaline benzenesulfonamide was active. These results provide a probably orientation in the binding site in relation to that for warfarin and lapachol.

Animals↗

Immunosuppressive properties of chloroquinoxaline sulfonamide.

Chloroquinoxaline sulfonamide (CQS), a sulfanilamide derivative with antitumor activity, was found to be toxic to lymphoid tissue during preclinical studies. The mechanism of this toxicity appears to involve profound inhibition of lymphocyte activation. Incubation of human peripheral blood mononuclear cells (PBMNCs) with CQS decreased cellular incorporation of thymidine and deoxyuridine in a dose-dependent manner. Analysis of cell cycle distribution by flow cytometry indicated that CQS blocked movement out of the G0/G1 phase. Drug-treated cells were smaller and expressed fewer receptors for interleukin-2 (IL-2) and transferrin than untreated mitogen-stimulated lymphocytes. These observations support the notion that CQS has cell cycle specificity in regulating lymphocyte proliferation. As little as 10 microM CQS markedly inhibited both human lymphocyte and murine CTLL cell replication in response to IL-2 containing growth factors. However, CQS did not block secretion of IL-2 into culture supernatant fractions by human PBMNCs. Finally, CQS inhibited in vitro production of immunoglobulins G and M by mitogen-stimulated lymphocytes, primarily by causing cytotoxicity. In all of these drug effects, CQS was approximately one to two logs more potent than the parent compound, sulfaquinoxaline (SQ). These studies indicate that CQS inhibits essential basic processes in human lymphocytes. This agent may find use as an immunosuppressive drug.

Animals↗

Are newer scientific concepts in regulatory toxicology used timely and appropriately?

Laws regulating toxicology (e.g. toxic thresholds allowed, poison classes or definition of necessary preclinical testing) might improve health and save lives. Scientific facts will always serve as a mandatory base for political decision-making, but there will also be additional influences (perception and acceptance of risks, possible benefits, economic considerations etc.). These latter factors may vary considerably from one society to another. The Delaney clause prohibited the marketing of any product which was found to be carcinogenic in animals. Due to their benefits, exceptions were made for drugs. In other countries, too, other chemicals could be an exception due to a different perception of the risk or different scientific evaluation. Clear cases of major events always trigger changes in legislation. When in 1937 a newly-marketed sulfanilamide elixir led to severe kidney damage and 70 deaths, the FDA quickly endorsed the propositions of the investigation team set up by the American Medical Association: animal testing in two species with histopathologic examination before a marketing authorization could be granted became mandatory. A similarly rapid reaction followed in Europe when it was detected that Thalidomide was responsible for malformations in the offspring of mothers who had taken the drug in early pregnancy. When the effects are more difficult to link to a chemical, there may be time delays in regulatory actions. However, a sophisticated evaluation system was introduced for better monitoring of drug and chemical hazards. Some examples will be given in order to discuss the difficulties of timely and appropriate use of scientific findings.

Animals↗

Carbonic anhydrase inhibitors: N-(p-sulfamoylphenyl)-alpha-D-glycopyranosylamines as topically acting antiglaucoma agents in hypertensive rabbits.

A series of N-(p-sulfamoylphenyl)-alpha-D-glycopyranosylamines was prepared by reaction of sulfanilamide with different monosaccharides in the presence of ammonium chloride. The new compounds were investigated for inhibition of the metallo-enzyme carbonic anhydrase (CA, EC 4.2.1.1), involved in aqueous humor secretion within the mammalian eye. Isozymes CA I and CA II were strongly inhibited by some of these compounds, which showed inhibition constants in the range of 510-1200 nM against CA I and 10-25 nM against CA II, similarly to clinically used sulfonamides, such as acetazolamide, methazolamide, dichlorophenamide, dorzolamide and brinzolamide. The presence of sugar moieties in these molecules induced an enhanced water solubility as compared to other sulfonamides. In hypertensive rabbits (a widely used animal model of glaucoma), two of the new compounds showed strong and long-lasting intraocular pressure (IOP) lowering, being more effective than dorzolamide and brinzolamide, the two clinically used, topically acting antiglaucoma sulfonamides with CA inhibitory properties.

Administration, Topical↗

Leishmania tarentolae: purification and characterization of tubulin and its suitability for antileishmanial drug screening.

Previously, tubulin has been purified from Leishmania amazonensis and used to identify novel molecules with selective antimitotic activity. However, L. amazonensis is pathogenic and requires a relatively expensive medium for large-scale cultivation. Herein, the purification and characterization of tubulin from the non-pathogenic Leishmania tarentolae is reported, together with the sequence of alpha- and beta-tubulin from this organism. This protein was purified by sonication, diethylaminoethyl-Sepharose chromatography, and one assembly disassembly cycle in 1% overall recovery based on total cellular protein. Leishmania tarentolae tubulin was indistinguishable from the corresponding L. amazonensis protein in terms of binding affinity for dinitroaniline sulfanilamides and sensitivity to assembly inhibition by these compounds. The amino acid sequences derived from the L. tarentolae alpha- and beta-tubulin genes were 99.6 and 99.4% identical to the corresponding amino acid sequences from the Leishmania major Friedlin strain. These results indicate that tubulin from L. tarentolae is suitable for use in drug screening.

Amino Acid Sequence↗