Clinical efficacy of acute respiratory viral infections prevention in patients with chronic heart failure.

AIM: The aim of the study was to evaluate the ARVI prevention effectiveness in patients with chronic heart failure (CHF) using interferon inducer amixin. MATERIALS AND METHODS: Conducted a comprehensive survey, dynamic monitoring and treatment of 60 patients aged from 49 to 70 years (mean age 60.25±4.57 years, 17 men and 43 women) with CHF with preserved ejection fraction of left ventricle (LVEF) (≥50%), II-III functional class (FC) according to the classification of new York Heart Association (NYHA), which developed as a result of coronary heart disease (CHD), hypertensive disease (HD). Of these, 30 patients (group 1) on the background of standard therapy for CHF received for the prevention of ARVI tiloron (Amixin) at a dose of 125 mg once a week for 6 weeks, two courses for 1 year. Group 2 patients received only standard therapy for CHF. RESULTS: A decrease in the frequency of ARVI in patients with CHF treated with Amixin was found, which was accompanied by a decrease in the severity of subclinical inflammation by reducing the production of proinflammatory (IL-1ß) and increasing the production of anti-inflammatory (IL-10) cytokines, reducing neurohumoral activation (reducing levels of aldosterone and Nt-proBNP), increasing the level of α- and γ-interferon. The positive dynamics of biomarkers of systemic inflammation and neurohormonal activation explains the improvement of the clinical course in patients with CHF (increase of tolerance to physical loads, reducing the number of visits to General practitioner and hospital admissions in the hospital during 12 months of observation). CONCLUSION: A promising approach to the prevention of SARS in patients with CHF is course therapy with Amixin (2 times a year before the seasonal rising in the incidence of respiratory viral infections and influenza), which allows to achieve both decreasing in the frequency of SARS per year, and improvement the clinical course of CHF.

Pulmonary administration of a dry powder formulation of the antifibrotic drug tilorone reduces silica-induced lung fibrosis in mice.

The aim of this work was to study the antifibrotic effect of pulmonary administration of tilorone to lung fibrosis. L-leucine coated tilorone particles were prepared and their aerosolization properties were analyzed using two dry powder inhalers (Easyhaler and Twister). In addition, the biological activity and cell monolayer permeation was tested. The antifibrotic effect of tilorone delivered by oropharyngeal aspiration was studied in vivo using a silica-induced model of pulmonary fibrosis in mice in a preventive setting. When delivered from the Easyhaler in an inhalation simulator, the emitted dose and fine particle fraction were independent from the pressure applied and showed dose repeatability. However, with Twister the aerosolization was pressure-dependent indicating poor compatibility between the device and the formulation. The formulation showed more consistent permeation through a differentiated Calu-3 cell monolayer compared to pristine tilorone. Tilorone decreased the histological fibrosis score in vivo in systemic and local administration, but only systemic administration decreased the mRNA expression of type I collagen. The difference was hypothesized to result from 40-fold higher drug concentration in tissue samples in the systemic administration group. These results show that tilorone can be formulated as inhalable dry powder and has potential as an oral and inhalable antifibrotic drug.

Development of sustained-release matrix tablets of BKP-01-041 (tilorone derivative) containing Hypromellose.

The objective of this research was to develop and evaluate sustained-release matrix tablets of BKP-01-041 (tilorone derivative) based on Hypromellose (hydroxypropyl methylcellulose, HPMC) as the matrix forming polymer. The sustained-release tablets were prepared by the wet granulation method. The influence of HPMC viscosity and ratios on drug release was investigated in vitro. Dissolution of the tablets developed with 26% HPMC K4 M/K100 M (1:2) (w/w) content showed a better drug release profile than the other batches tested in 12 h. Drug release from the optimal formulation was analyzed using release kinetics equations. The release kinetics parameters were determined and the value of the exponent (n) representing the apparent drug release mechanism determined from the Peppas equation was about 0.726. These results suggest that the drug release mechanism was non-Fickian (0.45 < n < 0.89), and drug release was dependent on both drug diffusion and polymer erosion.

[The role of immunomodulating therapy in the treatment and prevention of exacerbations of chronic cystitis].

The article presents the results of the examination and treatment of 60 women with chronic cystitis aged 20 to 80 years. The mean disease duration was 4,9 years (ranged from 6 months to 40 years). Immunomodulator Tilorone was included in the complex therapy of 30 patients of the main group. Besides the standard examination, immunological studies, including the definition of netrophil phagocytic rates in the patients were performed. It was revealed that the inclusion of immunomodulator in the treatment regimen of exacerbations of chronic cystitis contributes to an increase of index of activation of neutrophil phagocytic reserve, significant reduction in the frequency of exacerbations of the disease after the basic and preventive courses of treatment.

Performance of tiloronoxim and tilorone determination in human blood by HPLC-MS/MS: method validation, uncertainty assessment and its application to a pharmacokinetic study.

A highly sensitive and selective HPLC-MS/MS method is presented for the quantitative determination of tiloronoxim and its metabolite tilorone in human blood. An aliquot of 200 microl human blood was extracted with a mixture of chloroform/ethyl ether (1/2, v/v), using metoprolol as the internal standard (the IS). Separation was achieved on an Xterra MS C18 column (50 mm x 2.1 mm, 5 microm) with a gradient mobile phase of methanol/water containing 15 mM ammonium bicarbonate (pH 10.5). Detection was performed using positive MRM mode on a TurboIonSpray source. The mass transitions monitored were m/z 426.3-->100.0, m/z 411.3-->100.0 and m/z 268.3-->116.1 for tiloronoxim, tilorone and the IS, respectively. The method was fully validated using total error theory, which is based on beta-expectation tolerance intervals and include trueness and intermediate precision. The method was found to be accurate over a concentration range of 1-100 ng/ml for both compounds. The measurement uncertainty based on beta-expectation tolerance intervals was assessed at each concentration level of the validation standards. This method was successively applied to a pharmacokinetic study of tiloronoxim in healthy volunteers.

[Open population comparative randomized clinical trial of lavomax efficacy and safety in combined treatment of non-gonococcal urethritis].

A total of 40 patients with non-gonococcal urethritis (NGU) were divided into two groups. Twenty patients of group 1 received standard antibacterial treatment while 20 patients of group 2 received the same treatment plus an immunotropic drug based on tiloron (lavomax) in a course dose 1.25 g. Patients of group 2 had no recurrences while in group 1 recurrences were seen in 25% patients. Addition of lavomax resulted in clinical and etiological cure and activation of local (secretory IgA) and systemic (interferons alpha and gamma in blood serum) factors in anti-infection defense in NGU patients.

Elevation of efficacy of cancer vaccine combined with interferon and inducer of endogeneous interferon synthesis amixin.

AIM: To study in vivo efficacy of combined administration of cancer vaccine (CV), interferon (IFN) and inducer of endogenous IFN - amixin. MATERIALS AND METHODS: Sarcoma-37 cells were transplanted to female Balb/c mice. For the treatment, CV prepared from sarcoma-37 cells with the use of cytotoxic lectines from B. subtilis B-7025, murine IFN and amixin or their combinations were used. IFN production, content of circulating immune complexes and level of specific IgG antibodies in blood serum were determined by standard immunologic methods. RESULTS: Using solid form of sarcoma-37 it has been shown that introduction of IFN and amixin significantly elevated efficacy of vaccine therapy, in particular index of tumor growth inhibition reach 89.2% and 81.7%. Upon combined use of CV and IFN or CV and amixin (25 mg/kg) respectively. Significant prolongation of average life span of the animals treated with CV and IFN or CV and amixin (25 mg/kg) has been registered (up to 92.7 -/+ 10.4 and 95.0 -/+ 6.2 days respectively, vs 46.8 -/+ 1.5 days for control animals). CONCLUSION: Obtained results have shown expediency of the development of schemes for combined introduction of CV with exogenous IFN, and with inducer of endogenous IFN (amixin) for elevation of efficacy of vaccine therapy.

alpha7 nicotinic acetylcholine receptor agonist properties of tilorone and related tricyclic analogues.

BACKGROUND AND PURPOSE: The alpha7 nicotinic acetylcholine receptor (nAChR) has attracted considerable interest as a target for cognitive enhancement in schizophrenia and Alzheimer's Disease. However, most recently described alpha7 agonists are derived from the quinuclidine structural class. Alternatively, the present study identifies tilorone as a novel alpha7-selective agonist and characterizes analogues developed from this lead. EXPERIMENTAL APPROACH: Activity and selectivity were determined from rat brain alpha7 and alpha4beta2 nAChR binding, recombinant nAChR activation, and native alpha7 nAChR mediated stimulation of ERK1/2 phosphorylation in PC12 cells. KEY RESULTS: Tilorone bound alpha7 nAChR (IC(50) 110 nM) with high selectivity relative to alpha4beta2 (IC(50) 70 000 nM), activated human alpha7 nAChR with an EC(50) value of 2.5 microM and maximal response of 67% relative to acetylcholine, and showed little agonist effect at human alpha3beta4 or alpha4beta2 nAChRs. However, the rat alpha7 nAChR maximal response was only 34%. Lead optimization led to 2-(5-methyl-hexahydro-pyrrolo[3,4-c]pyrrol-2-yl)-xanthen-9-one (A-844606) with improved binding (alpha7 IC(50) 11 nM, alpha4beta2 IC(50)>30 000 nM) and activity at both human and rat alpha7 nAChR (EC(50)s 1.4 and 2.2 microM and apparent efficacies 61 and 63%, respectively). These compounds also activated native alpha7 nAChR, stimulating ERK1/2 phosphorylation in PC12 cells. CONCLUSIONS AND IMPLICATIONS: Tilorone, known as an interferon inducer, is a selective alpha7 nAChR agonist, suggesting utility of the fluorenone pharmacophore for the development of alpha7 nAChR selective agonists. Whether alpha7 stimulation mediates interferon induction, or whether interferon induction may influence the potential anti-inflammatory properties of alpha7 nAChR agonists remains to be elucidated.

[Allergenic and immunotropic properties of yeast RNA-tilorone hydrochloride molecular complex].

Immunotropic properties of the interferon-inducing molecular complex (MC) yeast RNA--tilorone hydrochloride have been under study. MC was experimentally studied in vivo to establish its influence on the amount of antibody-forming cells and the level of antibody formation. The influence of MC on the oxygen-generating activity of spleen macrophages was established in the HCT test. MC in a dose of 1.25 mg/kg was shown to considerably activate immunocompetent cells, thus producing pronounced influence on humoral immunity. In addition, the study showed the dose dependence of the influence of MC on individual elements of the immune system as well as differences in the dynamics of immunomodulation caused by the use of high and low doses of MC. The data thus obtained made it possible to regard MC as a promising immunomodulator.

[Individual changes of gene expression in the interferon system in human blood cells due to amixin and cycloferon].

The action of amixin and cycloferon on the expression of genes in the systems of interferon (IF) and cell apoptosis (CA) was studied by semi-quantitative RT-PCR in human blood microsamples before and after the administration of the drugs. Individual changes were determined in the transcription activity of genes of IF (alpha, beta, gamma), enzymes 2',5' oligoadenylatesynthetase (OAS), RNSase L, dsRNA-dependent proteinkinase (dsPK) and of CA effectors (FasAg, bcl-2, gamma-actin) registered dynamically in 24 h and 48 h. The activity parameters of IF genes were compared with the results of biological titration of IF activity in blood samples in vivo and in vitro. A pronounced ability of cycloferon to stimulate selectively the activity of genes of human IF, type I (beta IF--by 100 times and alpha IF--by 10 times), without affecting essentially the activity of other genes in blood cells, was detected. Amixin was found to inhibit the titration of genes with high activity levels. (alpha-, beta-IF, RNAases L, bcl-2 and gamma-actin). The antiviral and IF-induced properties of the drug are explained to a great extent by the apoptotic effect (activation of genes Fas, gamma-IF, OAS and affected transcription of gene bcl-2). A positive correlation was observed between the processes of activation of IF-genes transcription and the production of the total circulating IF. Antagonistic relations between type I and II IFs in human blood cells were shown.

[Antiviral effectiveness of the combined use of amixine and virasole in experimental hemorrhagic fever with renal syndrome in sucking albino mice].

The antiviral effectiveness of the combined and single use of superlow-dose amixine and virasole on the course of experimental hemorrhagic fever with renal syndrome was studied in sucking albino mice parenterally infected with their virus Hantaan. The co-administration of virasole and amixine was shown to protect 52% of the infected animals from death, which is superior to the effect of their monotherapy. The combined use of the drugs substantially prolongs the survival of albino mice after their infection and the level of brain viral reproduction suppression ( delta = 3.21 g) in the experimental group as compared to the controls and to the mice given only one of the drugs.

Pharmacokinetics of amixin after repeated peroral administration to mice.

Pharmacokinetics of amixin was studied after repeated administration (5 days) to animals. Perorally administered amixin is characterized by high bioavailability and is present in the circulation in high concentrations for a long time. The main pharmacokinetic parameters were estimated by the method of linear regression because of slow elimination of amixin from organs and tissues. Our results indicate that repeated treatment with amixin holds much promise for the prevention and therapy of chronic diseases (particularly hepatitides).

[The antirelapse effectiveness of amyxin used in combination with antiherpetic vaccine in patients with herpetic keratitis]. / Protivoretsidivnaia éffektivnost' amiksina v kombinatsii s protivogerpeticheskoi vaktsinoi u bol'nykh s gerpeticheskimi keratitami.

The efficiency of the combined use of amixin and anti-herpetic vaccination (AHV) to prevent the relapses of herpetic keratitis (HK) is investigated. One hundred and four patients with HK were followed up. The efficiency was found to be higher in the group of patients who received both amixin and AHV: in 29 (87.9%) patients with surface HK and in 10 (90.9%) patients with deep HK. No effect was registered in 4 (12.1%) persons with surface HK and in 1 (9.1%) person with deep HK. Amixin, when combined with AHV, cuts the number of relapses, therefore, it extends the remission period. The suggested scheme of amixin administration is as follow: 125 mg, once per week, for 10 weeks, 10 pills per one treatment course.

Correlations between in vitro effects of preparations of interferon and its inducers on blood cells in patients with multiple sclerosis.

We studied in vitro production of interferon-alpha and interferon-gamma by peripheral blood leukocytes from 15 patients with multiple sclerosis. The priming effects of interferon preparations weakly correlated with interferon-alpha production by leukocytes from patients with multiple sclerosis, but negatively correlated with interferon-gamma production. The effects of interferon inducers in most cases positively correlated with its spontaneous production. We found a weak positive correlation between the priming effect of natural interferon-alpha and the effect of recombinant interferons. There were positive or strong positive correlations between the effects of recombinant interferons on leukocytes from patients with multiple sclerosis. The relationship between the effects of medicinal interferon inducers and interferon preparation varied from negative to strong positive correlations. These data suggest that correlation analysis can be used for dynamic control and elaboration of methods for combined immunotherapy of multiple sclerosis with various interferon preparations or interferon and its inducers.

[Iodoantipyrine--an agent for treating and preventing tick-borne encephalitis]. / Iodantipirin--sredstvo dlia lecheniia i profilaktiki kleshchevogo éntsefalita.

The antiviral activity of iodantipyrine was studied in outbred [correction of inbred] albino mice (weight 10-12 g) infected with the Absettarov strain of the tick-borne encephalitis virus. Iodantipyrine was administered per os or parenterally and the animals were observed for 21 days. A reliable therapeutic effect was produced in 60% of mice infected with 10 DL50 of tick-borne encephalitis virus which were given the drug per os in a dose of 50 mg/kg. Preventive administration of the drug was effective in 47% of the animals.

Drug-induced glycosaminoglycan storage: dose-dependent changes in the pattern of accumulated glycosaminoglycans in cultured bovine and human fibroblasts.

The present study determines the amounts and patterns of glycosaminoglycans stored in cultured corneal fibroblasts after treatment with tilorone and three related compounds. The compounds have immunomodulatory properties and have been shown to impair the lysosomal degradation of glycosaminoglycans as a side effect. This side effect has been described as drug-induced mucopolysaccharidosis because the induced lysosomal storage of glycosaminoglycans leads to cellular lesions resembling those in patients with inherited mucopolysaccharidosis. In the present study, the dose-dependency of glycosaminoglycan storage was analyzed after treatment (96 hr) of bovine corneal fibroblasts. The investigated drug concentrations ranged from low concentrations inducing cytological lesions typical of drug-induced mucopolysaccharidosis to high concentrations at the borderline of cytotoxicity. The intracellular amounts of dermatan sulfate, heparan suflate, and chondroitin sulfate were quantified by densitometric scanning of Alcian Blue-stained bands after electrophoresis. All investigated compounds induced a predominant dermatan sulfate storage (3-4-fold accumulation) at low drug concentrations. With rising drug concentrations, a shift of the pattern of stored glycosaminoglycans was observed, characterized by the additional accumulation of heparan sulfate (up to 5-fold of control levels). In cultured human fibroblasts, tilorone also caused a marked dermatan sulfate storage, reaching maximum values at 5 microM and marked heparan sulfate storage at 20 microM. The present data provide evidence: (a) that selective dermatan sulfate accumulation is a characteristic feature of drug-induced glycosaminoglycan storage in cultured bovine and human fibroblasts, if these cells are treated with low concentrations (< or = 5 microM), that are likely to reflect the situation in vivo; and (b) that additional heparan sulfate storage is induced in vitro only by treatment with high concentrations that induce nonspecific cellular lesions.

Lysosomal storage of sulphated glycosaminoglycans induced by dicationic amphiphilic drug molecules: significance of the central planar ring system.

The immunomodulatory drug tilorone (2,7-bis[2-(diethylamino)ethoxy]fluoren-9-one) and several congeners are known to disturb the lysosomal degradation of sulphated glycosaminoglycans and thereby induce lysosomal storage of glycosaminoglycans in cultured cells and intact organisms. The molecules of tilorone and congeners consist of a planar aromatic ring system symmetrically substituted with two aliphatic side chains each carrying a protonizable nitrogen. In a previous study it was proposed that non-degradable glycosaminoglycan-drug complexes are formed by electrostatic interactions and that additionally intermolecular interactions between the drug molecules due to electronic coupling of their central planar ring system are important for formation and stabilization of the glycosaminoglycan-drug complexes and thus for the drug side effect in question. The significance of the central planar ring system was tested in the present study by comparing tilorone and the compound bis(beta-diethylamino-ethylether)hexestrol (DH) with respect to their potencies to cause lysosomal glycosaminoglycan storage in cultured bovine corneal fibroblasts. DH has the same side chains as tilorone, but its central apolar moiety lacks planarity. At a concentration (1.75 muM) which did not cause enhanced secretion of the lysosomal enzyme beta-hexosaminidase (E.C. 3.2.1.52), DH was significantly less potent than tilorone in causing storage of [35S]glycosaminoglycans. This is taken as support of the hypothesis that the planar tricyclic ring system is essential for the high potency of tilorone and its congeners to exert this adverse action.

Immunomodulation and drug acetylation: influence of the immunomodulator tilorone on hepatic, renal and blood N-acetyltransferase activity and on hepatic cytosolic acetyl coenzyme A content.

The biochemical alteration responsible for immunomodulator enhancement of drug acetylation in vivo was probed ex vivo and in vitro in the rat. Rat liver or kidney cytosol, obtained by differential centrifugation, or whole blood served as the source of N-acetyltransferase (NAT). Addition of tilorone (0.5-8.0 mM) to incubation mixtures containing procainamide (PA, 0.6 mM) and acetyl coenzyme A (AcCoA, 0.42 mM) resulted in the inhibition of N-acetylprocainamide formation, while lower concentrations of tilorone had no effect. Pretreatment of rats with tilorone (50 mg/kg) administered orally 48 hr prior to sacrifice did not alter hepatic apparent Km and Vmax for NAT toward PA compared to control animals. Utilization of an AcCoA regenerating system in the incubation mixtures also resulted in no significant differences in the apparent Michaelis-Menten parameters obtained. Acetylation activity in kidney and whole blood also was not altered by immunomodulator pretreatment. Hepatic cytosolic AcCoA content was reduced significantly 48 hr after tilorone pretreatment (5.10 +/- 2.1 vs 11.97 +/- 2.2 nmol/mg protein) (P less than 0.05). These data indicate that an increase in NAT content or activity is not the biochemical alteration responsible for immunomodulator enhancement of drug acetylation, and that the required cofactor, cytosolic AcCoA, is decreased by immunomodulator pretreatment.

[The comparative characteristics of the interferon-inducing capacity of 2 preparations related to aromatic hydrocarbons]. / Sravnitel'naia kharakteristika interferonindutsiruiushchei sposobnosti dvukh prparatov, otnosiashchikhsia k aromaticheskim uglevodorodam.

Aromatic hydrocarbons are rightly considered to belong to most active synthetic interferon inducers among low molecular compounds. A comparative evaluation of L-1 (acridanon) and amixin (fluorenon) showed L-1 to have more marked interferon-inducing properties. Both compounds differed not only in the dynamics and levels of interferon synthesis in different organs which suggests the possibility of their employment in different diseases, but also in the efficacy of the modes of application. L-1 induced IF synthesis most actively after subcutaneous inoculation, amixin after oral administration.

[Interferon-inducing activity of amixin and its effect on the interferon status]. / Interferonindutsiruiushchaia aktivnost' amiksina i ego vliianie na interferonovyi status.

The national preparation amixine induces interferon (IF) production in most of the people who have background values of serum IF. When the initial values of serum IF are high, after administration of amixine they decline to the background ones. A certain group of subjects (approximately 25%) is not sensitive to amixine. The IF content in the blood serum of such subjects does not change before or after administration of amixine. Its use should be limited to 4-5 administrations weekly, because its longer use leads to the exhaustion of the IF system in people.