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Studies on azopyrazole derivatives. Part 9: Synthesis and antibacterial evaluation of novel phenylhydrazones and their substituted 4-arylazopyrazole derivatives.

6-Methyl-2,4-heptandione (1) was coupled with diazonium salts of sulfanilamide (2) and 4-aminobenzoic acid (3). Resulting new hydrazones, namely 4-methyl-2-oxovaleraldehyde p-substituted phenylhydrazones 4 and 5, were refluxed with various hydrazines to synthesize the title compounds 6-15. Their structure elucidation was made on the basis of their analytical and spectroscopic data. The antibacterial activity was evaluated. Some of the compounds tested exerted moderate to potent antibacterial activity against Staphylococcus aureus, Staphylococcus epidermidis and Streptococcus pyogenes.

Anti-Bacterial Agents↗

[Preparation of monoclonal antibody against sulfadiazine and development of an ELISA kit].

AIM: To prepare monoclonal antibody (mAb) against sulfadiazine (SD) and develop an ELISA kit for rapidly detecting residual SD in different kinds of samples. METHODS: BALB/c mice were immunized with the conjugate of SD and BSA, and then anti-SD mAb was prepared by hybridoma technique. The purified ascitic mAb and HRP-labeled SD were used to establish a competitive ELISA for detection of SD in samples. RESULTS: 5 hybridoma cell lines secreting anti-SD mAbs 1A1, 1B8, 1E4, 2C1 and 3D9 were obtained. 1A1, 1B8 and 1E4 belonged to IgG1, while 2C1 and 3D9 belonged to IgG2. The I50 and theoretical minimum detectable amount of the kit was 9.3 microg/L and 0.6 microg/L, respectively. The recovery rates of the kit for SD in different kinds of samples were higher than 60%. The cross-reaction rate of the kit for other sulfanilamide drugs was lower than 3%. CONCLUSION: 5 mAbs against SD have been prepared successfully and possess high titer and specificity. The development of an ELISA kit for rapidly detecting SD can meet the needs of detection of SD in different samples.

Animals↗

Indirect UV detection in HPLC determination of UV-transparent non-electrolytes in pharmaceutical dosage forms.

After briefly reporting the basic concepts of indirect UV detection in HPLC, methods for the quantitative determination of ethanol, glycerol and isopropanol in pharmaceutical forms for human use, and propylene glycol and N,N-dimethylacetamide in pharmaceutical form for veterinary use are proposed. Nicotinamide, sulfanilamide and isoniazid are used as UV-absorbing components (probes) of mobile phase. The last two are new probes suitably selected and proposed to optimize the determination. Also, the dependence of method sensitivity from the probe concentration is shown. Good accuracies and precisions are obtained. The methods are simple and quick and the assays of the analytes do not go through interferences from the other substances present in the pharmaceutical forms.

Chromatography, High Pressure Liquid↗

[Rhinosinusogenic intracranial complications and their diagnosis at present].

Despite the use of strong antibiotics and sulfanilamides the frequency of rhino-sinusogenic intracranial complications do not have a tendency to decrease. Rather frequently this is explained by late diagnosis of sinusitis and their intracranial complications. Up-to-date methods of observation help to make both a thorough diagnosis of intracranial rhinosinusogenic complications and simplify the technique of differential diagnosis between brain abscess and meningoencephalitis.

Arachnoiditis↗

[On the plurifactorial determinism of the organophosphorous-induced teratogenesis on bird embryos; trials of protection by various compounds: oximes, hydroxamic acids and nicotinamide analogs (author's transl)].

Simple methods were applied to study the teratogenesis in Quail embryos induced by two important organophosphorous compounds: parathion and dicrotophos. Parathion led only to vertebral malformations, as other natural and synthetic cholinomimetics: nicotine, carbamylcholine, decamethonium, neostigmine... Dicrotophos induced not only vertebral malformations (specific to neuromuscular junction poisons) but also beak, legs and feather abnormalities (peripheric malformations which are also produced by insuline and sulfanilamide). Oximes and hydroxamic acids, some of these being analogs of nicotinamide, were tested as antiteratogens. The 3-(CO-NH2), or -(CO-NHOH), substituted pyridinic compounds (nicotinamide, nicotinohydroxamic acid) prevent perfectly dicrotophos-induced beak and legs malformations, in tertiary amine form, but very little in quaternary amine form (methyliodide). The 4-substituted pyridinic compound (isonicotinohydroxamic acid) and aliphatic oxo-oximes were quite ineffecient against these malformations. The vertebral malformations, as a rule, were not lessened by the compounds tested, except for isonicotinoyl-formaldoxime methyl iodide and in some degree for nicotinohydroxamic acid. From these observations, it results that teratogenesis induced by compounds as dicrotophos is rule by a plurificatorial determinism. The beak and legs malformations are prevented by analogs of nicotinamide. In the contrary, the vertebral malformations induced by parathion or dicrotophos are nicotinamide unsensitive and are only prevented by powerful cholinesterase reactivators as pralidoxime or TMB4 (MEINIEL, 1976 b) but are reduced little or not at all by less potent cholinesterase reactivators (HEATH).

Abnormalities, Drug-Induced↗

o-Phthalaldehyde based spectrophotometric determination of sulfonamides.

o-Phthalaldehyde (OPA) reacts with primary amino group in a compound in presence of a thiol (RSH) and in an alkaline medium to give a soluble product called isoindole. This is a fluorescent compound, yet it has been exploited for spectrophotometric study by measuring the absorbance at 340 nm which is the wavelength for excitation of the molecule. The spectrophotometric method has successfully been employed in the quantitative estimation of sulfonamides such as sulfadiazine, sulfanilamide and sulfamethoxazole, all containing a primary amino group. The detection ranges from 0.01 to 0.24 mg/ml of the active ingredient in different samples containing sulfonamides. The respective standard curves were linear in the given range and the relative standard deviation of the mean response factor ranged from 1.95-2.08%. 2-methyl-2-propane thiol (2MPT) has been used in the derivatization reaction for the formation of OPA-adduct and borate buffer of pH 10.0 has been used to maintain the medium alkaline.

Journal Article↗

[Integrons of bacteria isolated from aquatic environment].

Bacteria of the genus Pseudomonas, isolated from the water of the lakes Shira and Itkul (Republic of Khakassia, Russia) were shown to contain integrons of class 1 with gene cassettes, contained in the variable segment (sized 1 and 1.3 kb), were shown. Out of three detected integrons only one integron (in P. aeruginosa) included the sulfanilamide resistance gene contained in the 3'-conservative segment. The resistance of bacteria to kanamycin and ceftazidime was not seemingly linked with the presence of integrons. On the whole, the study revealed the presence of a significant proportion (27%) of integron-positive strains among aquatic bacteria with pronounced resistance to antibiotics.

Anti-Bacterial Agents↗

[Study on the TLC-SERS of sulfamethoxazole].

The spectra of sulfamethoxazole, the efficient ingredient of sulfanilamide-like medicine, were investigated by combining the TLC and FT-Raman spectroscopy using the surface enhanced Raman spectroscopy technique. The result indicatesthat the main vibrant characteristic spectral band can be obtained by TLC in a samples of just about 1 microg. The difference between the compound sulfamethoxazole and the corresponding spectra picture is analyzed. The analysis shows that the method of combining TLC and FT-Raman spectroscopy by the SERS is practical and advantageous in the highly sensitive measurement of the chemical ingredient in medicine.

Chromatography, Thin Layer↗

[Drug resistance of opportunistic enterobacteria isolated from various sources].

Resistance of 159 strains of opportunistic enterobacteria to 9 antibacterial drugs was studied. The strains were isolated from man and cattle. It was shown that the overwhelming majority of the isolates (93 per cent) were polyresistant irrespective of the genus. There was a high frequency of the strains resistant to the widely used antibiotics such as chloramphenicol (73 per cent), ampicillin (73.6 per cent) and rifampicin (95.6 per cent) and sulfanilamides (99.3 per cent). Gentamicin and nalidixic acid proved to be the most active against the cultures: 11.9 and 10 per cent of the resistant strains, respectively. The strains of enterobacteria isolated from different sources had a sensitivity to the antibiotics. Multiple antibiotic resistance to at least 5 drugs, variability of the spectra and high resistance were more characteristic of the isolates from the animals. The necessity of a rational use of antibacterial drugs in veterinary is indicated.

Animals↗

[Antimalarials and pregnancy].

Paludism can occur quite easily in pregnant women in endemic zones, above all those who are primiparous or in their 2nd or 3rd terms. The onset of paludal attacks can be serious for both the mother and the child. That is to say, besides the obviously imperative therapeutic action, a prophylaxis is also a necessity. The use of antipaludial substances at our disposal has been complicated during the last few years as a result of chloroquine-resistance extension. Besides a few nuances of kinetic nature observed in pregnant women, a good knowledge of teratogenous or embryotoxic effects is necessary. But this remains fragmentary. Among the principal antipaludial medications is quinine (Q), reported to be abortifacient but in reality it is not: it is often poorly tolerated by the mother (hypoglycemia), but is not responsible for abnormalities in children, except under large doses. Chloroquine (CQ), considered to be without harmful effects, can be used in women without large restrictions, even if toxic effects have been observed in animals. The pyrimethamine-sulfanilamide (P-S) combination contains two substances which are a potential risk. Nevertheless, experiments have never showed harmful effects in pregnant women, particularly when under cover of a joint prescription of folinic acid. Proguanil is without doubt the only molecule which can be used without restriction. Two new medications, quinoline methanol, Mefloquine (MQ) and Halofantrine (HF) are contra-indicated for lack of experimentation and because of some abnormalities observed at high doses in animals. Artemisinine and amino-8-quinoline are contra-indicated, and cyclines are strongly inadvised. From the practical point of view, the present use of antipaludial medication in pregnancy should take into account the surrounding risk, namely that of paludism and of treatments. Curatively, Q remains a serious treatment in any form. In CQ-sensitive zones CQ is usable unreservedly in simple attacks. In CQ resistance zones the use of Q seems preferable to that of Fansidar proposed by certain people. MQ and HF, although contra-indicated, have already been employed without inconvenience. By way of prevention, it is important first of all to avoid all leisure stays in endemic zones. If travel is unavoidable or for indigenous people, a chemoprophylaxis, judged according to the local risk of impaludation, is desirable: CQ in sensitive zones, PG+CQ in resistant zones, P-S, as proposed by some people, is normally contra-indicated; MQ and HF are contra-indicated. Protection against nocturnal mosquito bites is still strongly applied (Mosquito net, repellents, insecticides).

Animals↗

Transport of cyclosporin A in kidney epithelial cell line (LLC-PK1).

The characteristics of cyclosporin A transport have been studied in cultured kidney epithelial cell line LLC-PK1. The uptake of cyclosporin A by LLC-PK1 cells was time-dependent and reached a steady state at about 30 min. The initial uptake was saturable and was inhibited by cyclosporin A analogs, cyclosporin C and D and by verapamil, but not by metabolic inhibitors such as 2,4-dinitrophenol and rotenone. Cyclosporin A uptake as well as efflux was strongly dependent on temperature. The Arrhenius plot for cyclosporin A uptake was biphasic, whereas the Arrhenius plot for sulfanilamide uptake, which is transported by a simple diffusion, was linear. These results indicate that a specific mechanism is concerned in the transport of cyclosporin A in LLC-PK1, cells.

Animals↗

[Eosinophilic lesions of the lungs (case reports and review of the literature)].

Overall 75 references concerning the etiology, pathogenesis and the clinical picture of eosinophilic lesions of the lungs were analyzed. Sixteen patients with drug-induced disease were placed under the author's observation. Of these, 8 patients fell ill due to penicillin, 3 to tetracycline, 3 to sulfanilamide intake and alpha to intal inhalations. In 10 patients, pneumonia was characterized by high blood eosinophilia (from 20 to 53%, 37.5 +/- 2.7%) and in 6, it was of moderate nature (from 10 to 15%, 11.8 +/- 0.8%). According to the author's data, the immunopathological response coupled with suppression of the T lymphocyte system, inhibition of the enzymatic activity of leukocytes, dysproteinemias with hypergammaglobulinemia, reductions of the concentration of IgG underlie the disease pathogenesis. In the patients with high blood eosinophilia, including 4 with polysystemic lesions, the disease ran a grave course. The therapeutic effect could only be attained owing to the monotherapy with prednisolone. In the long-term period, 9 patients developed glucocorticoid-dependent bronchial asthma. The patients with moderate blood eosinophilia showed a milder disease course and a favourable outcome.

Adult↗

[Megaloblastic pancytopenia in a female patient with rheumatoid arthritis given methotrexate and amidopyrine simultaneously].

The paper is concerned with the development of megaloblastic pancytopenia in a female patient with grave rheumatoid arthritis. The patient had been given methotrexate (15 mg per week) for a long time, supplemented by amidopyrine (1.0 to 1.5 g/day). The author holds that the development of the above-indicated complication is provoked by both drugs interference into folate metabolism. It is concluded that caution should be taken as to the combined use of methotrexate with non-narcotic analgesic drugs, nonsteroidal anti-inflammatory agents and sulfanilamides.

Acute Disease↗

Kinetics and mechanism of degradation of some 2-sulfanilamidopyrimidine derivatives: part IV. Mechanism and kinetics of specific acid-catalyzed hydrolysis of sulfadimidine.

The following products of acid hydrolysis of sulfadimidine (SDMP) were isolated and identified: sulfanilic acid (Sac), sulfanilamide (SA), 2-amino-4, 6-dimethylprimidine (ADMP) and 2-hydroxy-4, 6-dimethylpyrimidine (HDMP). The apparent first-order rate constants (k1, k2 and k3) were determined for the reaction: (formula: see text). The catalytic rate constants kH+ = ki/[H+] and the thermodynamic parameters delta Ha, delta H not equal to, deltaS not equal to, deltaG not equal to and log A were also determined for that reaction.

Chemical Phenomena↗

Determination of inorganic nitrate in serum and urine by a kinetic cadmium-reduction method.

Nitrate in serum and urine was assayed by a modification of the cadmium-reduction method; the nitrite produced was determined by diazotization of sulfanilamide and coupling to naphthylethylene diamine. After samples were deproteinized with Somogyi reagent, the nitrate was reduced by Cu-coated Cd in glycine buffer at pH 9.7 (2.5 to 3 g of Cd granules for a 4-mL reaction mixture). The reduction followed pseudo-first-order reaction kinetics, a convenient time interval for assay being 75 to 90 min. Maximum reduction (85%) occurred at about 2 h. Detection limits in urine or serum were 2 to 250 mumol/L. This method does not require the reaction to go to completion, does not require expensive reagents or equipment, and can assay several samples simultaneously. Repeated assays of two serum pools gave CVs of 9.0% and 4.7% for nitrate concentrations of 31.4 and 80.2 mumol/L, respectively (n = 20 each). The mean concentration of nitrate was 1704.0 +/- 1294 (SD) mumol/L (n = 21) in untimed normal urine, 81.8 +/- 50.1 mumol/L in serum of 38 renal dialysis patients, and 51.2 +/- 26.4 mumol/L in serum of 38 controls.

Cadmium↗

Synthesis and antifungal evaluation of some 3-phenyl-2,5-disubstituted indoles derived from new ethyl-2-benzyl-2-[N-(aryl)hydrazono] ethanoates.

In connection with previous related works [1-6] we have now studied the reaction of ethyl alpha-benzyl-alpha-acetylethanoate (EBAE), as an aliphatic active C-H compound, with the diazonium salts of p-aminobenzoic acid (PABA), sulfanilamide (SA) and N,N-dimethylsulfanilamide (DSA), respectively. Ethyl 2-benzyl-2-[N-(aryl)hydrazono] ethanoates were the resulting substances (1-3). The synthesis of 4-6, namely 3-phenyl-2,5-disubstituted indoles, was carried out employing acidic indole ring closure reaction (Fischer indole synthesis) starting from 1-3. Because only 5 had a high antifungal potency its derivatives 7-10 were also prepared. But these derivatives were unfortunately found to be inactive. Structure elucidation of 1-10 were made by elemental analysis, IR, NMR and mass spectral data.

Antifungal Agents↗

Purification and characterization of human salivary carbonic anhydrase.

A novel carbonic anhydrase was purified from human saliva with inhibitor affinity chromatography followed by ion-exchange chromatography. The molecular weight was determined to be 42,000 on sodium dodecyl sulfate polyacrylamide gel electrophoresis, indicating that the human salivary enzyme is larger than the cytosolic isoenzymes CA I, CA II, and CA III (Mr 29,000) from human tissue sources. Each molecule of the salivary enzyme had two N-linked oligosaccharide chains which were cleaved by endo-beta-N-acetylglucosaminidase F but not by endo-beta-N-acetylglucosaminidase H, indicating that the oligosaccharides are complex type. The isoelectric point was determined to be 6.4, but significant charge heterogeneity was found in different preparations. The human salivary isozyme has lower specific activity than the rat salivary isozyme and the human red blood cell isozyme II in the CO2 hydratase reaction. The inhibitory properties of the salivary isozyme resemble those of CA II with iodide, sulfanilamide, and bromopyruvic acid, but the salivary enzyme is less sensitive to acetazolamide and methazolamide than CA II. Antiserum raised in a rabbit against the salivary enzyme cross-reacted with CA II from human erythrocytes, indicating that human salivary carbonic anhydrase and CA II must share at least one antigenic site. CA I and CA III did not crossreact with this antiserum. The amount of salivary carbonic anhydrase in the saliva of the CA II-deficient patients was greatly reduced, indicating that the CA II deficiency mutation directly or indirectly affects the expression of the salivary carbonic anhydrase isozyme. From these results we conclude that the salivary carbonic anhydrase is immunologically and genetically related to CA II, but that it is a novel and distinct isozyme which we tentatively designate CA VI.

Amino Acids↗

[Sensitivity of Yersinia pestis to antibiotics: 277 strains isolated in Madagascar between 1926 and 1989].

The antimicrobial susceptibility of 277 strains of Yersinia pestis was studied using broth microdilution panel. In recent strains, trimethoprim, cotrimoxazole and ampicillin were the most active of the antibiotics tested (MICs less than 2 mg/l). All strains were inhibited by 16 mg/l of kanamycin and sulfamethoxazole, 32 mg/l of sulphadiazine and sulfanilamide, and 64 mg/l of sulfamethoxypyridazine. Doxycycline, minocycline, chloramphenicol, demeclocycline, tetracycline, oxytetracycline and chlortetracycline were equally active but some of strains were resistant (13% for tetracycline; 32.5% for oxytetracycline; 84.5% for chlortetracycline). Analysis of MICs in relation with time (comparison of the two period: 1926-1948 and 1948-1989) evidenced a trend towards a decrease in susceptibility to cyclines prime generation. Sulphonamides and streptomycin preserve the some efficacity on Y. pestis but we have to notice the possible existence of resistant strain on high level with streptomycin.

Anti-Bacterial Agents↗