WO2001078751A1

WO2001078751A1 - Antiviral and antibacterial pharmaceutical preparation 'armenicum' and its use for treatment of infectious diseases

Info

Publication number: WO2001078751A1
Application number: PCT/AM2000/000002
Authority: WO
Inventors: Alexander Ilyin; Emil Gabrielyan; Levon Mkhitaryan
Original assignee: 'armenicum+' Jsc
Priority date: 2000-04-17
Filing date: 2000-11-24
Publication date: 2001-10-25

Abstract

The invention relates to the area of medicine, in particular to antiviral and antibacterial drugs and also their use in treatment of infectious diseases. The aim of the invention is to create a preparation that is effective against DNA- and RNA-containing viruses of mammals, including humans, and may be used for the treatment of viral and bacterial diseases, such as AIDS, herpes simplex, encephamyocarditis, etc. The essence of the invention is that the pharmaceutical preparation, besides iodine, potassium or sodium iodide, sodium chloride, lithium chloride, synthetic water-soluble polymer, mono-, oligo-, polysaccharides and water, additionally contains halogenderivatives of natural compounds, particularly halogenderivatives of mono-, oligo- and polysaccharides. The preparation has a low toxicity and is an effective means for treatment of AIDS and associated opportunistic diseases.

Description

Antiviral and antibacterial pharmaceutical preparation "Armenicum" and its use for treatment of infectious diseases

The invention relates to the area of medicine, in particular to antiviral and antibacterial drugs and also their use in treatment of infectious diseases.

Many drugs have been offered for the last three decades for viral diseases treatment. However, clinical trials of the drugs led to erroneous conclusions owing to the absence of correct assessment of the natural development in the majority of viral infections and missing knowledge about the affect of virus on living cells. Recent research advances in a more detailed study and development of tools for monitoring of virus infections, acquisition of biochemical knowledge about virus replication as well as a better understanding of the behavior of the virus in specific host cell populations have resulted in a tendency to develop more potent drugs to be used against viral infections. The discovery of endogenic interferon and its antiviral activity has turned out to be a milestone. At present, it has been established that the action of a number of immunostimulating and antiviral drugs is conditioned on their interferonogenic activity, i.e. the ability to stimulate the endogenic interferon formation [Mashkovsky M.D. Drugs. Manual for phisicians - Kharkov, "Torsing", 1998, v.2, p.349 (in Russian)].

Natural (human leukocytic) interferon relating to proteins is still being used as a prophylactic means against influenza and other viral infections.

There are also preparations containing aminoacids [DE 3811177; A 61 K 31/95; appl. 31.03.88; publ.19.10.89], peptides [EP 0545991; WO 92/03147; A 61 K 37/02; appl. 20.08.91; publ. 05.03.92] and modified plasma proteins [WO 90/0734; A 61 K 37/04; .appl. 30.01.89; publ.12.07.90] that display anti-HIN activity. However, these are all significantly inferior to interferon in activity.

From the recent most effective antiviral synthetic drugs we should note rimantadine, used for treatment of diseases caused by influenza virus, adenovirus, picornavirus and other viruses [Kui ain R.A., et al. Antiviral activity and mechanism of action of various chemical agents - Riga, "Znaniye", 1979 (in Russian)]. A dangerous feature of the viruses is their ability to mutate Epidemic viruses (Spanish Flu, Hong Kong Flu) and also human immunodeficiency virus (HIV) that belongs to retroviruses are considered to be animal viruses but they affect humans too

In treating AIDS a liposomic, lipid and polyene (e.g. nystatin and amphotericin B) containing pharmaceutical composition is used However, liposomic compositions show selective toxicity for HIV- 1 -infected cells [WO 92/118415, A 61 K 9/127, 31/71, 1992]

Various derivatives of glycyrrhizic acid have been offered as anti-HIV active compounds a fixed combination of glycopeptide of β- glycyrrhizic acid with glycyl-L-valyn methyl ester [RU Patent 2024542, C 07 J 53/00, 63/00, A 61 K 31/705, appl 04 03 91, publ 15 12 94], a fixed combination of amide β-glycyrrhizic acid with L-histidine methyl ether [RU Patent 2024543, C 07 J 53/00, 63/00, A 61 K 31/705, appl 04.03 91, publ 15 12 94], a fixed combination of amide β-glycyrrhizic acid with 6-aminouracil [RU Patent 2024545, C 07 J 53/00, 63/00, A 61 K 31/705, appl 04.03.91, publ. 15.12.94].

Rapamycin or its analogue is also used in ADDS treatment. It delays the development of HIV infection and slows down its spread in mammals [WO 94/05300, A 61 K 31/71, 1994]

Povidone-iodine is known to have a wider range of antiviral and antibacterial action as compared to other antiseptics. It is active against many virus types including HIV in suspension experiments in vitro [Kawana R et al., Inactivation of human viruses by povidone-iodine in comparison with other antiseptics - Dermatology, 1997, 195, Supll 2, pp 29-35] Povidone- iodine-containing anti-infective agents may be used in the treatment of HIV-, chlamydia-, gonococcus-, treponema pallidum- and herpes simplex virus-induced diseases [Tsutomu S , Junko S. Anti-infective against STD relating, for example, to HIV, chlamidia, gonococcus, treponema pallidum or herpes simplex and device used to preserve/mix principal ingredient and base of such anti-infective - EP 0823996A1, A 01 N 59/12, A 61 K 9/00, appl 15 08 96, publ 18 02 98 and Shanbrom E. Antiviral, spermicidal vaginal gel and foam containing low molecular weight povidone-iodine -US 5545401, A 61 K 31/79, A 61 K 33/18, appl. 02 06 94, publ.13 08 96]

It has been determined that compositions containing nonoxinol-9 oligomer, polyvinylpyrrolidone and iodine or polyvinylpyrrolidone-iodide have pronounced synergetic anti- HIV action as compared to the action of separate ingredients [Digenis G A , Digenis A G Coated products with potent anti-HIV and antimicrobial properties WO 95/28165, A 61 K 33/18, 9/10, appl. 18.04.94, publ 26 10.95] It has also been determined that the introduction of iodine into polyurethane polymers causes effective inactivation of HIV in contact with polymers [Shikani A.H , St Clair M , Domb A Polymer-iodine inactivation of the human immunodeficiency virus - J Am Coll Surg , 1996, 183(3), pp. 195-200].

The pharmaceutical preparation "Iodomidol" [KZ Patent 6730, appl 26 01 96, publ 16 11 98] has antibacterial and antiviral action and contains iodine, potassium or sodium iodide, a synthetic water-soluble polymer (playing the role of a polymer matrix), mixture of natural polymers, such as polysaccharides, mono- and oligo saccharides and water in the following proportions of ingredients, g/1

Iodine 6-10

Potassium or sodium iodide 9- 15

Synthetic water-soluble polymer 2-4

Mixture of mono-, oligo- and polysaccharides 8-120

Water the rest

This preparation has relatively high toxicity and is aimed exclusively at the treatment of animals' viral and other bacterial diseases.

There is also a pharmaceutical preparation with benanomicine A or B as an active ingredient for inhibition of HIV contamination [EP 0356330A, A 61 K 31/71, publ 15 09 90]

A number of side effects caused by the aforementioned preparations used for the inhibition of HIV and AIDS treatment is their common disadvantage.

There are two antagonist elements in pathogenesis of HIV-infection. the active, aggressive action of HIN and the protective response reactions of the organism [ΠOKPOBCKHH B B πaτor He3 n 3τκoτpoπHaH Tepaπiw BHH-HHφeKUHH 3πHfleMHθJioπιa H HHφeKunoHHbie 6oπe3HH, 1998, Ν5, c. 53-58.]

HTN replication cycle includes the following stages

• Attachment to host cell;

• Uncoating of virus;

• Control of DΝA, RΝA and/or protein production, • Assembly of virions,

• Release of virions,

Antiviral drugs may affect various stages of the virus replication cycle

The antiretroviral drugs may be classified according to their mechanism of action

1. Nucleoside analogue of reverse transcriptase (RT) inhibitors (NARTI) - zidovudine, didanosine, zalcitabine, lamivudine, stavudine,

2 Non-nucleoside RT inhibitors (NNRTI) - delavirdin, nevirapine, loviride,

3 Protease inhibitors (PI) - ritonavir, indinavir, saquinavir, nelfinavir

Routinely the drugs are administered in combination (2, 3 and 4 drugs) to reduce the possibility of developing drug resistance

The disadvantage of the above listed drugs, derivatives of nucleic acids that have to penetrate into the cell in order to display activity, is their significant negative influence on non-infected cells This results in the appearance of agranulocytosis, thrombocytopenia and erythropenia Because of toxicity, these drugs are frequently used only topically

The most similar to the claimed preparation in chemical composition and the achieved result is the pharmaceutical preparation [EP 0823996A1, A 01 N 59/12, A 61 K 9/00, appl 15 08 96, publ. 12.02.98 and AM Patent 659 A 61 K 33/14, 33/18, 31/70, 31/74, publ 1999], that shows antiviral action and is composed of iodine, potassium or sodium iodide, lithium chloride, synthetic water-soluble polymer, a mixture of natural mono-, oligo- and polysaccharides in the following proportions of ingredients, g/1

Iodine 0 8-25

Potassium or sodium iodide 1 2-38

Lithium chloride 0 1-20

Synthetic water-soluble polymer 0 01-6

Mixture of mono-, oligo- and polysaccharides 8-400

Water the rest

This preparation (with additional 9g/l physiological solution of sodium chloride ) may be used for the treatment of humans, but like the aforementioned, shows relatively high toxicity The invention is intended to create an antiviral and antibacterial drug lacking the above- mentioned disadvantages Another aim of the invention is to use the preparation for the treatment of HIV infection, as well as opportunistic viral and bacterial infections

An antiviral and antibacterial pharmaceutical preparation that contains iodine, potassium or sodium iodide, sodium chloride, lithium chloride, synthetic water-soluble polymer, mono-, oligo- and polysaccharides, water and additional halogenderivatives of natural compounds in the following proportions of ingredients g/1, is claimed

Iodine 0 8-25

Potassium or sodium iodide 1.2-38

Sodium chloride 9

Lithium chloride 0 1-20

Synthetic water-soluble polymer 0 01-6

Mixture of mono-, oligo- and polysaccharides 8-400 Mixture of halogenderivatives of natural 0 001-0 01 compounds

Water the rest

As halogenderivatives of natural compounds, the halogenderivatives of mono-, oligo- and polysaccharides are used.

Besides, the halogenderivatives of other natural compounds (vitamins, nucleosides, aminoacids) are known, which can be involved in the composition of the claimed preparation in case of having the ability to form water-soluble complex compounds with iodine

As synthetic water-soluble polymers with gel-forming properties are used the polymers (approved by the International Pharmacopoeia and the Pharmacopoeia of USSR, edition XI) forming water-soluble complexes with iodine in declared concentrations of iodine and other ingredients As polysaccharides are used all the natural polysaccharides, including antibiotics and their derivatives relating to this type of substances, which can form water-soluble complex compounds with iodine in declared concentrations of iodine and other ingredients,

The concentrations of the ingredients are strictly substantiated All the ingredients composing the claimed antiviral and antibacterial preparation are essential and efficient for the achievement of the set goal. No ingredient may be excluded or substituted for another, otherwise the desired result will not be achieved.

The biologically active ingredient of the preparation is iodine With a concentration below 0.8g/l, the reaction of iodine hydrolysis takes place

I₂ + H₂O → HI + HOI I

HI + O, illustrating, that the pharmaceutical preparation can not be stored over a long period of time

At a iodine concentration higher than 25 g/1, due to the formation of a iodine complex associates with other ingredients of the preparation, gel occurs which make the preparation unsuitable for medical use. Furthermore, in course of time, a possibility of a natural mono-, oligo- and polysaccharides halogenation occurs and the preparation's toxicity increases due to HI formation.

The concentrations of other ingredients have been selected in a way to provide reactions of a complex formation, association and aggregation in process of making the preparation in order to achieve antiviral and antibacterial action.

Routinely synthetic water-soluble polymers (polyvinyl alcohol, poly-N-vinylpyrrolidone) upon introduction into the blood perform the function of plasma expanders, sorbents (ex toxins) or prolongators. In the claimed preparation, a synthetic water-soluble polymer performs an important gel-forming function.

It is known that carbohydrates, in particular mono-, oligo- and polysaccharides together with proteins and lipids are the components of biological membranes that separate the cells from each other and their surroundings. Mono-, oligo- and polysaccharides that constitute this preparation are not alien elements to the organism of mammals, including humans

It should also be noted that one of the crucial points in the selection of the preparation ingredients is that the role of a matrix is performed by iodine complex compounds with low and high molecular ligands, which in their turn form associates Their aqueous solution has some gel properties

Iodine being a component of the preparation both in the molecular form and in the form of salts (potassium or sodium iodide), exists in several active forms formed as a result of complex formation reactions: I₂+ KI <→ KI₃ →- K + I₃ (1) nI₂ + L <→ L*nI₂ (2)

L-nI₂ <-» [L-I]⁺ + I ^.D (3)

where L is ligand (mono-, oligo- polysacchari.de and synthetic water-soluble polymer), n = 1 or 2

The association of complex compounds (according to reactions 1-4) leads to gel formation in the preparation.

Iodine and its various forms in the preparation may interact with almost all types of substances, which constitute the organism of mammals, including humans, and the structure of the cell and its membrane proteins, carbohydrates, lipids, amino acids, glico- and phospholipids, hormones, enzymes, vitamins, etc [Moroz J D , Nelson P Entropic elasticity of twist - storing polymers - Macromolecules, 1998, pp.1-37] Along with, the complex formation and halogenation reactions are taking place. However, the sequence of biochemical processes in the living organism is such that after the reaction of one of the active iodine forms a new active form of iodine emerges in other complexes, such as iodine-enzyme, iodine-hormone, iodine-biological membrane, iodine-protein, etc.

Each newly formed complex is biologically active and "native" for organism of mammals, including humans

Due to the mechanism of "recognition" and to specific chemical reaction, iodine (being an ingredient of the pharmaceutical preparation) as well as its complexes formed in the organism, halogenate (iodinate) the DNA- and RNA-containing viruses located both inside and outside the cell [Fujihashi T , Hara H , Sakata T., Mori K , Higuchi H , Tanaka A , Kaji H , Kaji A , Anti- human immunodeficiency virus (HIV) activities of halogenated gomisin J derivatives, new nonnucleoside inhibitors of HIV type 1 reverse transcriptase - Antimicrob Agents Chemother, 1995, v 39, No 9, pp 2000-2007]

Halogenation primarily proceeds through replacement of the hydrogen atom in the > N-H and -NH₂ groups of the purine and pyrimidine bases contained in the molecules of DNA- or RNA-containing viruses

\ \

N - H + I₂ ^»L → N - I + L'HI (5)

/ / H H

/ /

N + I «L → - N + L«HI (6)

\ \

H I

As a result of halogenation, new agents emerge, which do not possess the properties of a virus The antiviral action of the pharmaceutical preparation is based on this

The introduction of mono-, oligo- and polysaccharides halogenderivatives in the preparation is crucial Although only one of the polysaccharides' halogenderivative (antibiotic nucleocidine) has been isolated from the natural sources, such compounds are promising [Barnett J.E G - Adv Carbohydrate Chem , 1967, v 22, p 177, Hanessian S - Adv Chem , Ser , 1968, v 74, p 159, Szarek W A - Adv Carbohydrate Chem , 1973, v 28, p 225] as the insertion of halogen into a molecule often leads to the appearance of new biological properties

It has been determined that dextranes related to the polysaccharides, e g dextransulphate, have a wide range of antiviral action [Witvrouw M , DeClercq E - Gen Pharmacol 1997, Nol.29, pp. 497-511] The mechanism of antiviral action of dextransulphate is the inhibition of retrovirus' (HIV) interaction with corresponding cell receptors [Watson K , Gooderham Ν. J., Davies D S., Edwards R.J - Biochem Pharmacol , 1999, Vol 57, pp 775-783]

At the same time, it has been asserted that some polysaccharides on the cell's surface play an important role in intracellular "recognition" Polysaccharides possess the potential ability of great structural polymorphism Thus, a group of carbohydrates on the cell's surface may have numerous variants, since a) monosaccharides may bind with each other through any of their hydroxyl groups b) binding on C-l may be both of α- and β-configuration, and intensive branching out of the chain is possible [Streier L Biochemistry - Translation from English - Moscow "Mir", 1984, v 1, p.215 (in Russian)]

Halogenderivatives administered m smaller quantities than dextransulphate because of their stronger ability to react will analogously interact with receptors on the surface of HIV-infected cells, T-lymphocytes [Jagodzinski P P , Wierzbicki A , Wustner J , Kanelo Y , Kozbor D - Viral Immunol , 1999, Vol 12, pp 23-33]

Thus, an additional possibility is created to influence an infected cell with complex compounds of the preparation formed in the result of its synthesis (reactions 1-4) Halogenids of alkaline metals introduced into the composition of the preparation play a non- typical role. They improve conductivity in cell membranes, enhance lateral diffusion in the of cell membranes due to the small size of metal cation and its more firm co-ordination on donor atoms of mono-, oligo- and polysaccharides.

The claimed preparation is a dark-violet aqueous solution of molecular and ion iodine complexes with associates of synthetic water-soluble gel-forming polymers and natural mono-, oligo- and polysaccharides. The melting point of the pharmaceutical preparation is -0.9 to -1.7°C, density 1.0374-1.1257 g/cm³ and pH 5-7.

The method of obtaining the claimed pharmaceutical preparation is the following: 1.2g of potassium or sodium iodide is dissolved with 100ml of water, add 0.8g of crystalline iodine and the solution is stirred until complete iodine dissolution. Separately, O.Olg of polyvinyl alcohol is dissolved with 100ml of water. Separately 8.0g of mono-, oligo- and polysaccharides mixture is dissolved in 200 ml of water. The obtained solutions are mixed in the following sequence: first the solution of mono-, oligo- and polysaccharides mixture and then the solution of iodine and potassium iodide are added to the solution of polyvinyl alcohol. After adding 9.0 g of sodium chloride and O. lg of lithium chloride, bring the solution to 11 with distilled water. Finally, the halogenderivatives of natural mono-, oligo- and polysaccharides are added.

The reactions of halogenation aimed to obtain halogenderivatives of natural mono-, oligo- and polysaccharides are performed in conformity with classic halogenation reactions of these compounds [Edwards R.G., Hough L., Richardson AC, Tarelli E. -Carbohydrate Res., 1974, v.35, p.lll;. Bhatt R.S, Hough L., Richardson A.C. - Carbohydrate Res., 1976, v.49, p.103; Foster A.B., J.H. Westwood - Pure Appl. Chem., 1973, v. 35, p.147], Identification of the obtained compounds is performed by the methods of elementary analysis and lR-spectroscopy (IR-spectrometer "1600 Series FTIR", Perkin-Elmer). In IR spectra of mono-, oligo-, and polysaccharides halogenderivatives an intensive absorption band C-X (X is I or Br) appears at 510 cm^*1 area.

For other concentrations of the ingredients of the claimed preparation the solutions are obtained according to above-described methods and in the same sequence

The compositions of the preparation and some of their physic-chemical characteristics are presented in Tables 1 and 2. The study of antiviral action of the preparation was carried out on cell cultures, mice and chick embryos.

In the experiments, influenza virus type A, strain A/Aichi 2/68 H₃N_2. human herpes simplex II virus, mice encephalomyocarditis virus and poliovaccine were used.

To support viability of viruses and to study the antiviral activity of the preparation, the passaged cell lines of human larynx adenocarcinoma (Hep-2), human rhabdomyosarcoma (RD), as well as primary cultures of chick embryos' fibroblasts were used. The cells were cultivated in the Eagle medium with glutamine and bovine serum (Hep-2, RD) and in 199 medium with serum of chick embryos' fibroblasts.

The known antiviral preparations rimantadine, acyclovir and γ-interferon were used as positive control in the experiments with influenza and herpes viruses.

The preparation has been tested in various conditions of the experiment: administration before and after contamination, and simultaneous administration with the virus.

The virus titration in the mice was performed by Rid and Mench method, in the chick embryos - according to hemaglutination reaction, and in cell cultures - according to the extent of cytopathic effect inhibition.

The antibacterial activity of the preparation was studied in compliance with conventional methods: by qualitative-suspension method, using 2 billion of microbial bodies in 1ml suspension and 250 million of microbial bodies in 1ml suspension.

Standard cultures: E. coli (strain 1257), enteropathogenic E. coli Crimea "0 151", St. aureus (strain 906), St. aureus 209 p., B. cereus (strain 96), B. cereus var. mycoides (strain 12), Mycobacterium B 5 and chloraminresistant cultures S. typhimurium, isolated from the patients(strain 5644), (strain 27), S.typhi (strain 33), Y. enterocolitica (strain 38), Ps. aeruginosa (strain 39), B. cereus thuringiensis (strain 40), B. cereus thuringiensis var. israelensis, V. cholerae El-Tor "Ogava"- 21 culture (strains 124-144).

The results were statistically evaluated in compliance with the European Pharmacopoeia and biometrical methods [Urbach B. Biometrical methods, M., 1964; GF XI USSR, p.199-251].

The results of laboratory experiments of the claimed preparation are presented in Tables 3-18.

Tables 3-6 present the data of the preparation's effect on influenza and herpes simplex viruses (HSV). As it follows from the presented data, the antiviral action of the preparation is comparable with the action of known preparations such as rimantadine and acyclovir. The study of the preparation's action on encephalomyocarditis virus (EMV) was carried out in vitro and in vivo. The data in Tables 7-9 show that the introduction of preparation both before and after contamination significantly suppresses the development of the virus at a contamination dose of 100 TCA₅o in vitro and results in prophylactic action in vivo.

Tables 10-12 present the results of the preparation's action on poliovirus. According to these data, the preparation completely neutralizes poliovirus reducing 10⁶ TCA₅o/ml titre up to zero that provides evidence of the preparation's virucidal action.

Tables 13-18 show the data regarding the study of bactericidal action of the preparation. The given data demonstrate a wide spectrum of bactericidal activity of the preparation, including activity against antibioticoresistant and chloramineresistant microorganisms, and may be employed as an efficient means against many nasocomial and wound infections.

The antiviral and antibacterial preparation may be used for the treatment of infectious deseases of mammals, including humans, particularly HIN-infection, as well as a number of opportunistic deseases.

The clinical trials were conducted in the volunteers in accordance with GCP requirements [Good Clinical Practice - John Wiley & Sons, 1998].

Pharmaceutically effective amount of the preparation is administrated intravenously. Before injection it is diluted with physiological solution of sodium chloride^" in 1 :4 to 1:30 v/v that makes its use convenient depending on the particular features of patients. The preferred speed of injection is 4-9 ml/min. Intramuscular, intraperitoneal, subcutaneous and other applications of the preparation are also possible.

Tables 19-31 summarize the results of clinical trials of claimed preparation. Table 19 presents data about 152 participants of clinical trials, segregated according to injected doses.

In Table 20 the alteration of ranged subjective and objective status is presented. The data are presented as Z-value (p), Wilcoxon matched-paired test [Wilcoxon matched-paired test, Armitage P., Berry G.: Statistical Methods in Medical Research (2^nd edition). Blackwell Scientific Publications, Boston, 1987, pp. 410-411, Conover W.: Practical Νonparametric Statistics (2^nd edition), New York; John Wiley & Sons, 1980]. The data in Table 20 provides evidence of statistically reliable improvement of subjective and objective status as compared to the status before the administration of the preparation. An obvious positive effect is observed at the first measurement of the status on the 12^th week and does not alter within a year. The data in Table 21 illustrate the statistically reliable weight gain in the 12^th week in the groups with 0.2 and 0.3 ml kg dosing. The following weight changes in the 24^th, 36^th and 48^th weeks are insignificant.

The data in Table 22 show that maximal decline of RNA viral load in plasma of patients in the group with 0.2 ml/kg takes place on the 8^th and 12^th week (by 1 08 and 1 02 log respectively)

The data of Table 23 show an alteration of RNA viral load in plasma of the examined patients with maximal initial values of viral load (>30.000 copies/ml) At that, statistically reliable decline of viral load is documented at 0 2 ml kg dose in the 8^th and 12^th weeks (by 1 078 and 1 019 log respectively).

The data in Table 24 show that the patients with a dose of 0 2ml/kg have statistically reliable increase of CD4+ on the 4^th, 8^th and 12^th weeks and the patients with a dose of 0 3 ml/kg - during the whole period of observation.

The data of Table 25 show, that the patients with a dose of 0 3 ml/kg have statistically reliable increase of CD4+ and CD8+ ratio on the 4^th and 12^th weeks (by 0 3 and 0 4 respectively).

As it is seen in Tables 26-28, the alteration of the parameters is similar in all cases of the preparation dilution with physiological solution in 1 4 - 1 30 v/v

Clinical trials prove that within the period of treatment with the preparation the biochemical and clinical parameters of plasma are approaching normal values

Quality of life measurements were carried out with the Medical Outcome study HIV Health Survey (MOS HIV Health Survey) validated tool specially designed for the measurement of the soft clinical endpoints of HIV-infected patients [Scott-Lennox J A , Wu A W , Boyer J G , Ware J E J. Realibility and validity of French, German, Italian, Dutch and UK English translation of Medical Outcomes Study FflN Health Survey - Med Care, 37, 908-25] The tool determining quality of life consists of 11 dimensions. The analysis of test results presented in Table 29 allows to conclude that patients treated with preparation have an obvious improvement of the quality of life in all dimensions

Tables 30 and 31 show that the use of the claimed pharmaceutical preparation results not only in the decline of RΝA HIV viral load and decrease of RΝA and DΝA HIV establishment but also in decrease of opportunistic viruses (Herpes Simplex type 1 and II and Cytomegalovirus) Meanwhile no additional preparations were used for the treatment of opportunistic infections

Clinical trials of the preparation and various methods of treatment in HIV-infected and AIDS patients allow to include the claimed preparation in the list of effective antiretroviral drugs An additional positive property of the preparation is its ability to affect opportumstic infections and essentially improve the patients' quality of life

The obtained data provide evidence that introduction of halogenderivatives of mono-, o go- and polysaccharides into the composition of the claimed preparation increases its efficiency as an antiviral and antibacterial pharmaceutical preparation

Toxicological studies were carried out in compliance with the current normative documents [Requirments to preclinical study of general toxic effect of new agents Methodic recommendations, Pharmacological Cimmittee of MH USSR, Moscow, 1985 The rules of preclinical asssessment of medical products' safety, Good Laboratory Practice Guiding regulatory document, Moscow, 1992]

Toxicological investigations of the antiviral preparation, conducted with laboratory ammals by the different routes of administration provide evidence of its relatively low toxicity and weak cumulative ability The preparation in therapeutic doses and in doses several times exceeding the middle day dose adjusted for humans, was tπaled several times with the animals The results have demonstrated that the preparation does not essentially influence the general state of health, behavior or temperature of the animals

In Tables 32-34 the acute toxicity parameters of the preparation are presented The presented data show the preparation's low toxicity at all routes of administration

A possible toxic activity of the preparation on cell cultures was determined in 1 10 - 1 80 dilutions (composition according to example 1 and 2) and 1 30 - 1 240 (composition according to example 3) The incubation of the monolayer of cell culture with the preparation was performed during 10, 30 and 60 minutes The extent of non-specific, cytotoxic action (CTA) was assessed according to the conventional 4-cross system The data obtained are presented in Tables 35-40 The Tables show that the preparation demonstrates lack of toxic effect in all dilutions the cells preserve normal morphology and viability At the same dilutions the preparation is not toxic for chick embryos and mice either

Thus, the results of the experiments on the antiviral action of the preparation obtained in vitro and in vivo, lack of toxicity, both prophylactic and therapeutic efficiency, the wide range of antimicrobial action (including antibioticoresistant and chloramineresistant microorganisms) prove its efficiency as an antiviral and antibacterial medicinal product Table 1

Composition of pharmaceutical preparation

Table 2

Phisic-chemical characteristics of pharmaceutical preparation

Table 3

Influence of the prophylactic administration of the preparation on outcome of the experimental influenza virus in mice

(composition according to example 2)

Table 4

Therapeutic effect of the preparation on influenza virus in mice

(composition according to example 2)

Table 5

The action of the preparation on HSV reproduction in chick embryos

(composition according to example 2)

Table 6

Statistical analysis of the antiviral effect of the preparation against HSV

(composition according to example 2)

Table 7

Influence of the various doses of the preparation on the reproduction of encephalomyocarditis virus (EM V) in the cells of Hep-2 cultures

(composition according to example 2)

Note: Numerator - number of test tubes with CPE; denumerator - number of contaminated test-tubes

Table 8

Influence of the preliminary and the subsequent administration of the preparation on the reproduction of EMV

(composition according to example 2)

Note: numerator - number of test tubes in CPE, denumerator - number of infected test tubes Table 9

Protective effect of the preparation in experimental encephalomyocarditis

(composition according to example 2)

Table 10

Antiviral action of the preparation on poliovirus

(composition according to example 1, dilution 1 2 v/v)

Numerator- number of test-tubes with CPE, denumerator - number of contaminated test tubes

Table 11

Antiviral action of the preparation on poliovirus

(composition according to example 2)

Note numerator- number of test tubes with CPE, denumerator - number of contaminated test tubes

Table 12

Antiviral action of the preparation on poliovirus

(composition according to example 3, dilution 1 120 v/v)

Note * - dilution of the preparation in 1 120 ratio, numerator - number of test-tubes with CPE, denumerator - number of the contaminated test tubes Table 13

Antibacterial action of the preparation

(composition according to example 1)

Note: >30 - preparation does not cause bactericidal activity within 30 minutes

Table 14

Antibacterial action of the preparation

(composition according to example 2)

Table 15

Bactericidal action of the preparation

(composition according to example 3)

Table 16

Influence of the preparation on microorganisms

(composition according to example 1)

Note "-" - presence of bactencidal action, "+" - lack of bacteπαdal action,

* - expeπments on Vibr choierae El-Tor were conducted on 20 strains, isolated from the patients

(the results were identical) All the expeπments were conducted following 10-πunute exposure Table 17

Influence of the preparation on microorganisms

(composition according to example 2)

Note: "-" - presence of bactencidal action, "+" - lack of bactencidal action,

*- the expenments on Vibr choierae El-Tor were conducted on 20 strains, isolated from patients

(the results were identical) All the expenments were conducted followmg 10-nunute exposure

Table 18

Influence of the preparation on microorganisms

(composition accordmg to example 3)

Note: "-" - presence of bactencidal action, "+" - lack of bactencidal action, * - the expenments on Vibr choierae El-Tor were conducted on 20 strains, isolated from the patients (the results were identical) All the expeπments were conducted followmg 10-nunute exposure Table 19

Table 20

Alteration of the ranged subjective and objective status

^* - statistically reliable alteration of subjective and objective status

** - the value of the following week is subtracted from the value of 0 week rangs sum Table 21

Alteration in mean body weight of volunteer participants in clinical trials during 48 weeks

^* - statistically reliable alteration of subjective and objective status

** - the mean weight of the participants in the following week of the clinical tπal is subtracted from the mean weight of 0 week participants

Table 22

Mean values alteration of viral load at different dosage

the value of the following week is subtracted from the 0 week value Table 23

Alteration of viral load mean values (common logarithm) (initial values of viral load >30 000 copies/ml)

* - statistically reliable alteration of viral load

** - the value of the following week is subtracted from the 0 week value

Table 24

Alteration of CD4+ in 24 weeks according to dosing groups

Table 25

Alteration of CD4+/ CD8+ ratio according to dosing groups

*- statistically reliable alteration of CD4+ and CD8+ ratio

" - the value of CD4+ and CD8+ ratio on the 0 week is subtracted from the CD4- and

CD8+ ratio value of every following week

Table 26

Results of treatment of HIV-infected and ADDS patients at use of preparation preliminary diluted with physiological solution in 1:4 volume ratio (by example of 3 patients)

^■ number of volunteers

Table 27 Results of treatment of HIV-infected and AIDS patients at use of preparation preliminary diluted with physiological solution in 1:10 volume ratio (by example of 3 patients)

• number of volunteers Table 28 Results of treatment of HIV-infected and AIDS patients at use of preparation preliminary diluted with physiological solution in 1:30 volume ratio (by example of 3 patients)

number of volunteers

Table 29 Quality of life in patients with measurements of viral load (111 participants)

I. Overall Health

* - the value of the average score at following measurements (3,6,9,12,15 months) is subtracted from the value of the average score before administration of the preparation (0 month)

H. Physical function

HI. Role function

IV. Social function

VU. Mental health

Table 30

DNA- and RNA HIV dynamics at administration of the preparation

* - the data are presented as: correlation of the DNA detecting πsks compaπng to 0 week (95%- confidence interval) ** - statistically reliable ratio of the risks

Table 31

Dynamics of Herpes Simplex type I and H and Cytomegalovirus pathogens establishment according to weeks (%, number of observations - 151)

Table 32

Parameters of the acute toxicity of preparation

(composition according to example 1)

* non toxic

Table 33

Parameters of the preparation acute toxicity

(composition according to example 2)

Table 34

Parameters of the preparation acute toxicity

(composition according to example 3)

Table 35

Determination of the preparation toxicity on RD cell culture at different periods of incubation

(composition according to example 1)

Note: numerator -lack of cytotoxic action (CTA); denumerator - number of test-objects

Table 36

(composition according to example 2)

Note: numerator -lack of CTA; denumerator - number of test-objects.

Table 37

(composition according to example 3)

Table 38

Determination of the preparation toxicity on Hep-2 cell culture

(composition according to example 1)

Note: numerator -lack of CTA, dnumerator - number of test-objects

Table 39

Determination of the preparation toxicity on Hep-2 cell culture

(composition according to example 2)

Note: numerator -lack of CTA; dnumerator - number of test-objects

Table 40

Determination of the preparation toxicity on Hep-2 cell culture

(composition according to example 3)

Note: numerator -lack of CTA, dnumerator - number of test-objects

Claims

1 Antiviral and antibacterial pharmaceutical preparation containing iodine, potassium or sodium iodide, sodium chloride, lithium chloride, synthetic water-soluble polymer, mono-, oligo- and polysaccharides and water, characterized in that it additionally contains halogenderivatives of natural compounds in the following proportion of ingredients,

Iodine 08-25

Potassium or sodium iodide 12-38

Sodium chloride 9

Lithium chloride 01-20

Synthetic water-soluble polymer 001-6

Mixture of mono-, oligo- and polysaccharides 8-400 Mixture of halogenderivatives of natural 0001-001 compounds

Water the rest

2. The pharmaceutical preparation according to Claim 1, characterized in that halogenderivatives of mono-, oligo- and polysaccharides are used as halogenderivatives of natural compounds

3. The use of pharmaceutical preparation accordmg to Claims 1 or 2 in treatment of mammals, including humans, infected with viral and bacterial infections

4 The use of pharmaceutical preparation according to Claim 3, characterized in that pharmaceutically effective amount of the preparation is administered intravenously

5 The use of pharmaceutical preparation according to Claim 4, characterized in t h a t the preparation is preliminarily diluted with physiological solution of sodium chloride from 1:4 to 1.30 volume ratio.

6 The use of pharmaceutical preparation according to Claims 4-5, characterized i n t h a t the preparation is administered at a speed of 4-9 ml/min

7. The use of pharmaceutical preparation according to Claim 3 characterized in that pharmaceutically effective amount of the preparation is administered intramuscularly, intraperitoneally, subcutaneously or orally

8 The use of pharmaceutical preparation according to any of Claims 3-7 character ized in that the preparation is applied for HIV-infection treatment

9 The use of pharmaceutical preparation according to any of Claims 3-7 character ized in that the preparation is used for treatment of AIDS and associated opportunistic diseases.