Eczema vaccinatum treated with n-methylisatin beta-thiosemicarbazone.
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A new mathematical method based on 3-, 4- or 6-point parallel-line assays was applied for working out a computer programme used to evaluate the activity of some new isatin-beta-thiosemicarbazone derivatives against vaccinia virus, studied in vivo.
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In view of earlier work which suggested that certain derivatives of isatin beta-thiosemicarbazone might be of possible clinical use in smallpox infections, several such derivatives have been tested for antiviral effect in infant mice inoculated intracerebrally with up to 10(3) LD(50) (up to 10(6.5) pock-forming units) of the Harvey strain of variola major virus.Survival of 100% was obtained in mice treated twice daily with N-methylisatin beta-thiosemicarbazone in a dose level of 10 mg/kg, isatin beta-thiosemicarbazone in a dose level of 25 mg/kg and 7-methylisation beta-thiosemicarbazone at 10 mg/kg, the treatment beginning immediately after infection. No evidence of toxicity was observed at these dose levels. At lower dose levels of these and other related compounds complete protection was not obtained, but a chemotherapeutic effect was demonstrated by a prolongation of the mean survival time. Mice which had survived variola infection after treatment with an adequate dose of chemotherapeutic agent were immune when challenged with 10(5) pock-forming units of rabbitpox virus.
In vitro experiments studied the antiviral activity of 11 different drugs against viruses of bovine infective rhinotracheitis (BIRT) and bovine viral diarrhea (BVD). The 50% inhibiting concentrations of the test agents were determined in the monolayers of MDBK and KCT cell cultures. Only did phosprenyl show a virucidal activity against BIRT virus. All the tested drugs significantly inhibited the reproduction of BIRT virus in the sensitive MDBK cell cultures. Thus, bromuridin, acyclovir, ribavirin and methisazonum inhibited the virus by > or = 100,000 times; liposomal ribavirin, gossypolum, anandinum, polyprenolum, phosprenyl, by 1000-10,000 times; eracond and argovit, by 100 times. In experiments on BVD virus, the cultured KCT cells displayed the antiviral activity of bromuridin, phosprenil, polyprenolum, methisazonum, acyclovir, gossypolum, argovit, and ribavirin (in two variants), which caused a statistically significant (100-10,000-fold) decrease in the productive activity of this virus. Eracond and anandid proved to be ineffective.
N,N'-bis(methylisatin-beta-thiosemicarbazone)-2-methylpiperazine in a 100 mumol/l concentration inhibited the reproduction of vaccinia virus in RK-13 cells by about 90%. This compound (bis-IBTMP) had no influence on virus adsorption and on early stages of virus multiplication, but affected virus reproduction from 12 to 24 hr post-infection (p.i.). The incorporation of 3H-thymidine into infected cells increased during first 10 hr p.i., decreasing gradually afterwards. In the infected cells treated with bis-IBTMP the same tendency was observed up to 10 hr p.i., but later on the incorporation level remained unchanged. The uptake of 14C-amino acids in the presence of bis-IBTMP was reduced both in vaccinia virus-infected and non-infected RK-13 cells.
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Experimental data of the evaluation of the antiviral activity of virazole in tissue culture using the fluorescent antibody technique are presented. Administration of virazole in the maintenance medium in a concentration range of 800--6.25 microgram/ml immediately after vaccinia virus adsorption exerts a marked inhibiting effect reducing virus reproduction by 3.1--1.6 1g, respectively. The chemotherapeutic index of the drug is 128. In a one-cycle experiment, virazole in the minimal effective concentration had the same inhibiting effect as metizason in the maximal tolerable concentration.
The effect of the compound N,N'-bis/methylisatin-beta-thiosemicarbazone/-2-methylpiperazine (bis-MIBTP) on immune response in BALB/c and Swiss mice have been studied in the course of vaccinia virus infection. Humoral response tested by neutralization and hemagglutination inhibiting antibodies was similar in compound-treated mice to this of untreated mice. Cell-mediated immune response, examined by spleen lymphocytes migration inhibition test, has been delayed or temporally depressed in bis-MIBTP treated mice as compared with the control group. High protective activity of the compound in vaccinia infected mice in spite of impairment of cellular immunity may indicate that antibodies have played an important role in recovery process from vaccinia infection.