WO2012091589A1

WO2012091589A1 - The use of viburnum opulus or its tissues or products thereof for the prevention or treatment of herpes virus infections

Info

Publication number: WO2012091589A1
Application number: PCT/PL2011/050057
Authority: WO
Inventors: Andrzej Lipkowski; Bogumiła LITWIŃSKA
Original assignee: Instytut Medycyny Doswiadczalnej I Klinicznej Pan; Narodowy Instytut Zdrowia Publicznego Państwowy Zaklad Higieny
Priority date: 2010-12-30
Filing date: 2011-12-29
Publication date: 2012-07-05
Also published as: PL221522B1; PL393515A1

Abstract

We claim the use of preparations of Viburnum opulus fruit in the prevention and therapy of HSV infections. The preparations exhibit a multifactor activity, both by depressing virus replication in infected cells, as well as via a protective effect on cells, thereby lowering their sensitivity to HSV infection. For this reason they may be used both in the prophylaxis of virus infections as well as in the therapeutic treatment of extant HSV infections. The properties of the preparations enable them to be used as the active ingredients of ointments or creams for external use as well as an active ingredient of pills for oral administration.

Description

THE USE OF VIBURNUM OPULUS OR ITS TISSUES OR PRODUCTS THEREOF FOR THE PREVENTION OR TREATMENT OF HERPES VIRUS INFECTIONS

A significant role in the etiopathogenesis of viral infections is played by viruses of the family Herpesviridae. In human, they cause a number of disease syndromes, from common dermal- mucous aberrations through to life -threatening infections of the central nervous system. These viruses cause infections which are among the most frequent infections in humans on all of the continents. The lack of effective immuno-prophylaxis against HSV means that the only currently available treatment for herpes virus infections is chemotherapy. The preparation used on a broad scale against HSV-1 infections is acyclovir, which was discovered in the 1970s. This is a guanosine analogue which induces the termination of DNA synthesis. The presence of a viral enzyme, thymidine kinase, is required for the activation of acyclovir, wherefore it is only slightly toxic to healthy cells. A very significant question in antiviral therapy is the formation of strains resistant to drugs used. It is supposed that along with the popularization of antiviral preparations this problem may become more significant, as was the case with antibiotic therapy. And although strains of HSV-1 resistant to acyclovir are most often less pathogenic, they pose a serious threat to patients with depressed immunity, particularly those with developed AIDS in whom long-term antiviral therapy may lead to the selection of resistant strains. According to literature data, the occurrence of HSV-1 strains resistant to acyclovir is about 5% in persons with depressed immunity, and as much as 14% among bone marrow transplant recipients (Nugier F, Colin JN, Aymard M, et al, Occurrence and characterization of acyclovir-resistant5 herpes simplex virus isolates: report on a 2-year sensitivity screening survey. J Med Virol 36 (1992) 1-12). The increasing number of persons with disrupted immunity and, particularly those with AIDS and transplant recipients behoves the intensive development of chemotherapies for the treatment of infections caused by HSV.

Throughout the world, many plant preparations have been used in unconventional medicine as agents against viral infections, recently, a number of substances isolated from plant extracts have been described, exhibit antiviral propeties against HSV-1 (Cheng H-Y, Yang C- M, Lin T-C, et al. (2006). Ent-Epiafzelechin-(4a~8)-epiafzelechin extracted from Cassia javanica inhibits herpes simplex virus typ e 2 replication. J Med Microbiol 55, 201-206, Kuo YC, Lin LC, Tsai WJ, et al, Samarangenin B from Limonium sinense suppresses herpes simplex virus type 1 replication in Vera cells by regulation of viral macromolecular synthesis. Antimicrob Agents Chemother 46 (2002) 2854-2864). An example of such a substance is n-docosanol, which was aproved for use against cold sores (Pope, LE, Marcelletli JF, Katz LR, et al., Anti-herpes simplex virus activity of n-docosanol correlates wit h intracellular metabolic conversion of the drug. J Lipid Res 37 (1996) 2167- 2216). These discoveries indicate that substances isolated from plants may be a future alternative to currently used chemotherapy.

The goal of the present invention is to design a novel drug for the prevention or treatment of herpes virus infections as well as to indicate a component which may be used in the manufacture of such a drug.

The goal of the present invention is to use Viburnum opulus or its tissues or products thereof in the production of a drug for the prevention or treatment of herpes virus infections. Preferably, in the production of the drug, use is made of Viburnum opulus fruit or products thereof, particularly fruit juice or its extract, preferably lyophilisate. Preferably, fruit extract at a concentration of from 0.5% to 5.0% by mass of the preparation is used in the production of a drug in the form of a preparation for external use, particularly a cream, ointment or lotion, whereas in the production of a drug in the form of a preparation for oral administration fruit extract is used at a concentration of from 0.5% to 50% by mass of the preparation.

As a result of experiments on Viburnum opulus it unexpectedly turns out that the fruit extract exhibits antiviral activity against the herpes virus (HSV-1). This research demonstrates the very selective activity of this extract against the mechanism of infection of HSV. At the same time, it was shown that the preparation has a multi-component activity, and both depresses the replication of the virus in infected cells, as well as exhibiting protective activity for cells, thereby decreasing their sensitivity to HSV infection. The properties of the preparation make it possible to use the active components as an ointment or cream for external use, as well as the active ingredient of tablets for oral administration. To better illustrate the subject invention based on the antiviral activity of preparations of Viburnum fruit, we attach examples which demonstrate the inhibitory activity of said preparations on HSV replication in cell cultures, under different variants of cell infection. The scope of the present invention should not, however, be limited to the following examples. Example 1

Juice was pressed from Viburnum opulus fruit, which was subsequently filtered through a tissue filter. The filtrate was lyophilised. The resulting dry preparation was used for subsequent investigations. A constant culture derived from monkey kidneys, HSV-1 infected CV-1 cells, was maintained, incubated for 1.5 h at 37°C; Subsequently, maintenance medium was added which contained 0.5% or 1 % of the dry Viburnum opulus fruit extract. In parallel, cultures were maintained under identical conditions without an addition of the plant extract. After 24 h and after 48 h the virus was collected and its infective titre was ascertained. The resulting data shown in the table show that a strong antiviral activity was obtained from the preparation of Viburnum opulus (a decrease by over 100 000 infective viruses in 1 ml)

Table 1. Virus titre (average values, two replicants).

Example 2

The preparation was obtained according to Example 1.

A culture HSV-1 infected CV-1 cells was maintained, incubated for 1.5 h at 37°C; Subsequently, maintenance medium was added which contained 0.5%> or 1 % of the Viburnum opulus fruit extract preparation. After 24 h and after 48 h the viruses were collected and their infective titre was ascertained. (Like in example 1 a decrease by over 100 000 infective viruses in 1 ml was obtained).

Table 2. Virus titre (average values, two replicants).

Example 3.

The preparation was obtained according to Example 1.

A mature CV-1 cell culture was depleted of its growth medium and the evaluated preparation solution was added at a concentraion of 0.5% or 1 % in maintenance medium. The culture was incubated for 2 hours, the evaluated preparation solution was removed and the culture was rinsed with maintenance medium, MEM-A. Next the culture was HSV-1 infected, incubated for 1.5 h at 37°C and after incubation, MEM-A was added. The viruses were collected after 24 h and after 48 h and their infective titre was determined.

Table 3. Virus titre for individual variants (average values, two replicants).

Infective titre value

Culture without the preparation

after 24 h 10-^{4 2} TCIDso

after 48 h 10-^{4 1} TCIDso

Culture with 0.5% of the preparation

after 24 h 1 0-^{3 25} TCIDso

after 48 h 10-^{2 9} TCIDso

Culture with 1.0% of the preparation

after 24 h 10-^{3 4} TCIDso

after 48 h 10-^{1 0} TCIDso

Claims

1. The use of Viburnum opulus or its tissues or products thereof in the production of a drug for the prevention or treatment herpes virus infections.

2. The use according to Claim 1, characterised in that the production of a drug makes use of the fruit of Viburnum opulus or products thereof, particularly the juice of its fruit or an extract thereof, preferably a lyophilisate.

3. The use according to Claim 1 or Claim 2, characterised in that in the production of a drug in the form of a preparation for external use, particularly a cream, ointment or lotion, makes use of a fruit extract at a concentration of from 0.5% to 5.0% by mass of the preparation.

4. The use according to Claim 1 or Claim 2, characterised in that in the production of a drug in the form of a preparation for oral administration makes use of fruit extract at a concentration of from 0.5% to 50% by mass of the preparation.