Inhibition of HIV infection of H9 cells by chlorpromazine derivatives.

The binding between the HIV surface protein, gp120, and the CD4 coreceptor is known to be initiated by electrostatic interactions. Because of the ability of chlorpromazine to interact with proteins by charge transfer, we tested several derivatives for their ability to block binding of HIV to CD4+ cells. We have shown that 7,8-dioxo-chlorpromazine blocks binding of fluorescein isothiocyanate-labeled anti-Leu3a and rgp120 to peripheral human blood T4 cells and blocks syncytia formation between gp120- and CD4-expressing cells. We also found that 7,8-dioxo-chlorpromazine blocks HIV infectivity of H9 cells and acts synergistically with zidovudine.

Effect of phenothiazines, benzo[a] phenothiazines, benz[c]acridines and Pentaglobin on endotoxin.

The endotoxin neutralizing effects of several phenothiazines, benzophenothiazines and Pentaglobin (control) were investigated by spectrophotometry, tumor necrosis factor (TNF) induction and the conventional Limulus test. In animal experiments, some beneficial effects of complex forming compounds were found, however, the compounds could not completely inactivate the biological effect of endotoxin in the Limulus test. The complex formation between endotoxin and the compounds were revealed in spectrophotometry. The TNF inducing effect of compound-endotoxin complexes was markedly reduced by some phenothiazines and benzo[a]phenothiazines in leukocytes. Benz[c]acridines and Pentaglobin could not neutralize completely the TNF induction of E. coli endotoxin. The recent findings indicate that multifocal effects of phenothiazines and benzophenothiazines can be responsible for anti-endotoxin action in vivo. Hypotensive action in experimental animals was reduced by some phenothiazines in some preliminary experiments.

Antiplasmid activity: loss of bacterial resistance to antibiotics.

The antiplasmid activity of tricyclic compounds, e.g. phenothiazines, dibenzoazepines, dibenzocykloheptene derivatives and some stereoisomers, was shown on E. coli in vitro. Some ring-substituted phenothiazine and cannabis derivatives had only an antibacterial effect. Promethazine, a selected phenothiazine, cured antibiotic resistance and lactose fermentation of E.coli, tumour inducing ability of Agrobacterium tumefaciens and nodule formation of Rhizobium meliloti. Plasmids of different E.coli strains were eliminated with varying frequency. The antiplasmid activity of the compounds can be due to the increased membrane permeability. Inhibition of DNA gyrase and complex formation with the supercoiled form of plasmid DNA can lead to the cessation of plasmid replication in the bacterial cells. In addition, in vivo plasmid curing was demonstrated at a low frequency.

The influence of charge transfer complex formation on the antibacterial activity of some tricyclic drugs.

1. Phenothiazines and structurally related drugs are effective in eliminating bacterial plasmids. 2. Charge transfer complex formation between chlorpromazine and xanthine dyes with different electron acceptor activities (e.g. fluorescein, eosin, erythrosin and rose bengal) was detected by differential spectrophotometry. Charge transfer complexes were formed between the strong electron acceptor rose bengal and various tricyclic drugs. 3. On the basis of the wavelength shift, the binding energies of drugs and dyes were estimated. 4. The temperatures dependence of the reaction indicates charge transfer complex formation rather than a chemical reaction between drugs and dyes. 5. The anti-plasmid action of the phenothiazines was decreased in the presence of xanthine dyes. As a consequence of competitive inhibition between bacterial binding sites and xanthine dyes, the binding energy of drugs in the plasmid replication system could be determined in the presence of dyes. 6. Drugs with binding energies in the range of 0.23-2.31 kcal/mol can inactivate the plasmid replication system.

Comparative study of antiproliferative effects of chlorpromazine, 7,8-dioxochlorpromazine, amantadine-N-mustard, rutin-N-mustard and alpha, beta and gamma interferon on K-562 cells in vitro.

The effects of rutin-N-mustard, amantadine-N-mustard, chlorpromazine and human interferon types alpha, beta and gamma (IFN-alpha, -beta and -gamma) were studied on the DNA, RNA and protein synthesis of K-562 cells. Monocyte-mediated cytotoxicity and immune spleen cell activity were examined in the presence of the same compounds (except for IFN-beta). The natural killer (NK) cell activity was tested in the presence of the two chlorpromazine compounds and the two N-mustard derivatives. Only 7,8-dioxochlorpromazine exerted an inhibitory effect on DNA synthesis. The protein synthesis of the cells was inhibited in the presence of IFN-alpha, -beta and -gamma. 7,8-Dioxochlorpromazine exerted some inhibition on both NK and immune spleen cell activity, while monocyte-mediated cytolysis was not altered. IFN-alpha, -beta and -gamma activated the cytolytic activity of monocytes and the NK activity in control experiments. Chlorpromazine, rutin-N-mustard and amantadine-N-mustard were ineffective in both tests in vitro. Rutin-N-mustard, 7,8-dioxochlorpromazine and the interferons may be assumed to have quite different antiproliferative mechanisms of actions.

Investigation of phenanthroquinolizidine analogues. Synthesis and biological activity of substituted 9-sec-amino- and tert-aminophenanthrenes.

A series of 9-tert-aminomethylphenanthrenes substituted with various groups in various positions were prepared, and their fungicidal effects were examined. Comparative studies were made with phenanthroquinolizidines, which are strong fungicides. Opening of the quinolizidine skeleton was found to be accompanied by a substantial decrease in the effect. This is considered to be connected with the change in position (orientation) of the nitrogen relative to the phenanthrene skeleton.

The effect of rutin-N-mustard on the survival of NK/Ly ascites tumour-bearing mice.

A newly synthetized rutin derivative, rutin-N-mustard, showed a marked antiproliferative effect on NK/Ly ascites tumour cells in vivo. When administered i.p. or i.v., this compound increased the survival time of the animals, but oral administration had no such effect. The drug was not able to kill the tumour cells, since the chromium release from the rutin-N-mustard-treated tumour cells was the same as in the case of the controls. Tumour growth was completely inhibited in the animals when the cells were pretreated with 100 mg/kg rutin-N-mustard before transplantation. The results also showed that rutin-N-mustard has little effect on the xenogenization of NK/Ly tumour cells. We presume that the antiproliferative action of rutin-N-mustard is due to its alkylating activity.

Drug-receptor interaction on plasmid elimination by phenothiazines and imipramine in Escherichia coli.

Plasmid elimination in Escherichia coli by a quaternary amine of chlorpromazine was demonstrated on different incompatibility groups of plasmid. The biological effect of the drug depends partly on the host bacteria and partly on the plasmid itself. Various receptor substrates such as adenosine, dopamine, histamine and norepinephrine do not alter the plasmid elimination by promethazine and imipramine. None of the known drug-receptors studied are involved in drug binding of the bacteria. The direct membrane action of imipramine and promethazine was demonstrated in electron microscopic studies and alterations in the bacterial membrane such as discontinuities, phase separation or rarely extensive lytic alterations were observed. Magnesium ions prevent the ultrastructural membrane alterations caused by imipramine and promethazine. There is some evidence that the drugs bind to two different receptor sites simultaneously on the plasmid replication site. The first and strongest binding has to be ionic through the side chain amino group, displacing the bivalent cations. In turn, the two aromatic rings of the fixed (ionically bound) drug molecules bind weakly through pi-electrons, hydrophobically or by a charge transfer complex. This weaker binding together with the ionic one are essential for biologic action and lead to the inhibition of plasmid replication. A schematic model of the effect of tricyclic psychotropic drugs on the bacterial membrane is proposed.

The effect of ring-substituted and quaternary chlorpromazine derivatives on the survival of the NK/Ly ascites tumor bearing mice.

The effect of quaternary chlorpromazines like-methiodide, allylammonium chloride, -octylammonium iodide, benzylammonium chloride, -phenacylammonium bromide, 9-phenanthryl-methylammonium bromide, trimethylene-bis-chlorpromazine-ammonium bromide and among the ring-substituted derivatives the 7-hydroxy-, 7,8-dihydroxy-, 7,8-dioxo-, 6,9-dihydroxy-, 6,9-dioxo-chlorpromazines and the chlorpromazine-5-oxide on the prolongation of survival time of NK/Ly ascites tumor bearing mice was studied. The intraperitoneally administered quaternary chlorpromazine derivatives in 10-25 mg/kg daily dose did not increase the life span of the animals, and did not prevent the development of the ascites tumor. However some ring system substituted chlorpromazines such as the 7,8-dioxo- and 6,9-dioxo-derivatives markedly increased the survival time of the tumor bearing mice and reduced the ascite volume in 10 mg/kg body weight dose.

Antitumor activity of flavonoids on NK/Ly ascites tumor cells.

Among the various flavonoids, rutin and quercetin increased the survival time of mice inoculated with NK/Ly ascites tumor cells. The best results were obtained when the mice were given 2.0 mg rutin twice daily for 8 days. The O-silyl-substituted rutin and quercetin were less effective than rutin or quercetin themselves. Besides rutin, quercetin and morin two other flavonoids, luteolin and pelargonidin also exerted growth inhibitory effects on NK/Ly ascites tumor cells cultures in vitro.