WO2012020370A1

WO2012020370A1 - Galvinoxyl derivative for use thereof against lipoprotein-enveloped viruses, in particular herpesviruses

Info

Publication number: WO2012020370A1
Application number: PCT/IB2011/053543
Authority: WO
Inventors: Robert Vachy
Original assignee: Robert Vachy
Priority date: 2010-08-10
Filing date: 2011-08-09
Publication date: 2012-02-16
Also published as: EP2603229A1; FR2963736B1; FR2963736A1

Abstract

The subject matter of the present invention is a galvinoxyl derivative chemically known as 2,6-di-t-butyl-(3,5-di-t-butyl-4-oxo-2,5-cyclohexadien-1-ylidene)-p-tolylhydroxy and represented by formula (I), for use thereof in the treatment of diseases caused by lipoprotein-enveloped viruses (LEVs), and in particular with respect to the herpesviruses. Said galvinoxyl derivative can furthermore be associated with propolis.

Description

GALVINOXYL DERIVATIVE FOR ITS USE AGAINST LIPOPROTEIC ENVELOPE VIRUSES, IN PARTICULAR HERPES VIRUSES

The present invention relates to a new therapeutic composition comprising a derivative of galvinoxyl alone or possibly associated with propolis, to a process for the preparation of said composition, and to the use of this composition as an antiviral means, in a manner general vis-à-vis lipoprotein enveloped viruses (abbreviated LEV), and in particular vis-à-vis herpes viruses and their analogues that are part of the set of LEVs.

Propolis is a gummy substance that is recovered in hives. Specifically, it is a substance that bees collect on certain plants, including poplar bud scales and alders, collect and use to seal hives cracks, fix the rays and varnish the walls. In the field of beekeeping, propolis is known as the antibiotic or disinfectant means of the hive preventing the proliferation of bacteria and mold.

Crude propolis generally contains resins and balsamic substances (approximately 55-50% by weight), beeswax (approximately 30-35% by weight), ethereal oils (approximately 10% by weight) and pollen (approximately 5% by weight), see in particular EP-A-0 061 508 (page 2, lines 1-28), EP-A-0 109 993 (page 1, lines 24-26) and EMSCHNEIDEWIND et al, Die Pharmazie 34, 103, (1979). EP-A-0 061 508 (page 3, lines 6-7) and EP-A-0 109 993 are known in particular.

(page 2, lines 3-8, page 10, lines 7-11, and page 11, lines 1-4) above, on the one hand, and EP-A-0 310 757 (page 3, lines 31-32 ), on the other hand, the antiviral properties of propolis extracted from hives and purified vis-à-vis herpes viruses (including HSVi and HSV ₂ ) and influenza viruses (including IV _A ).

For convenience, the following abbreviations are used in the present description.

EBV = Epstein Barr Virus

HSVi = herpes simplex virus type 1

HSV ₂ = herpes simplex virus type 2 HSViR = herpes simplex type 1 resistant to acyclovir [reference antiviral with the structural formula of 2-amino-1,9-dihydro-9 - [(2-hydroxyethoxy) methyl] -6H purine-6 one]

IV _A = influenza virus type A

LEV = lipoprotein envelope virus

Galvinoxyl derivative (DEG)

Prp = propolis

Rp = weight ratio

RT = room temperature (15-20 ° C)

TV = infectious titer of the sample of

ZV = Zoster virus

According to the invention, it has surprisingly been found that a galvinoxyl derivative can be used as an antiviral medium for lipoprotein enveloped viruses (LEVs), and in particular vis-à-vis 'herpes.

Thus, the present invention more particularly relates to a galvinoxyl derivative chemically named "2,6-di-t-Butyl- (3,5-di-t-butyl-4-oxo-2,5-cyclohexadien-1-ylidene "p-tolylhydroxy" and represented by the following formula:

for its use in the treatment of diseases caused by lipoprotein enveloped viruses (LEVs).

For convenience, 2,6-di-t-Butyl- (3,5-di-t-butyl-4-oxo-2,5-cyclohexadien-1-ylidene) -p-tolylhydroxy compound as defined herein above will simply be referred to as "galvinoxyl derivative" in the present application. According to an advantageous embodiment of the invention, the galvinoxyl derivative as defined above is more particularly used in the treatment of diseases caused by the herpes HSVi, HSV ₂ and / or HSViR strains. According to another advantageous embodiment of the invention, the galvinoxyl derivative as defined above is associated with propolis.

The present invention also relates to a pharmaceutical composition characterized in that it comprises a galvinoxyl derivative as defined above.

According to another advantageous embodiment of the invention, the composition as defined above further comprises propolis.

If this is the case, said composition comprises from 100 to 650 parts by weight of the galvinoxyl derivative for one part by weight of propolis.

The composition of the invention as defined above is used in the treatment of diseases caused by lipoprotein-enveloped viruses (LEV), and more particularly in the treatment of diseases caused by herpes HSVi, HSV ₂ strains. and / or HSViR.

The composition as defined above comprises the galvinoxyl derivative in combination with one or more excipients suitable for oral, injectable or local administration.

This composition may further comprise propolis.

As mentioned above, such a composition will be particularly advantageous vis-à-vis herpes and herpes-like diseases. It has indeed been found that such a composition is particularly effective in the treatment of herpes simplex type 1 and type 2, especially diseases caused by the HSVi and HSViR strains, which are transmissible by contact between mucous membranes. on the one hand, and diseases caused by the HSV ₂ strains, which are sexually transmitted and include, in particular, venereal growths, on the other hand. According to the invention, it is recommended a new therapeutic use according to which the derivative of galvinoxyl alone or possibly associated with propolis is used for obtaining an antiviral drug intended for use in human or veterinary therapy vis-à-vis diseases caused by viruses lipoprotein envelope, including diseases caused by strains (i) HSVI, _HSV2 and HSViR and (ii) IVA, EBV and ZV.

The antiviral composition according to the invention may be prepared according to a method known per se.

The process for the preparation of the composition is characterized by mixing the galvinoxyl derivative alone or with propolis in a suitable aqueous or oily physiological medium at a temperature of between 15 ° C and 75 ° C.

When the galvinoxyl derivative is associated with propolis, the weight ratio of the galvinoxyl derivative to propolis ranges from 100/1 to 650/1.

Other advantages and features of the invention will be better understood in the following reading of comparative tests and examples of preparation. Of course, all of these elements is in no way limiting but is given by way of illustration.

EXAMPLE 1 - Comparative Tests -

A culture medium containing DeG at concentrations of 50 or 25 mg / ml and solutions of Prp at decreasing concentrations of 0.363 mg / ml, 0.272 mg / ml, 180 mg / ml, 136 mg / ml, is used. 0.090 mg / ml. Equal volumes of said culture medium and said solutions are incubated in the presence of HSViR for 1 hour at 37 ° C with shaking. After centrifugation, the infectious titres are determined in the usual manner according to the method known as the limiting dilutions and compared to control samples of HSViR virus, propolis control samples and control samples of HG1 incubated under the same conditions (it has not It has not been necessary to prepare control samples of excipients since previous tests have demonstrated the lack of effect of the excipient on viruses, especially HSViR). To precisely measure the interaction of DeG and Prp, the method described by RFSCHINAZI et al., Antimicrob, was used. Agents Chemother. , 22 (No. 3), pages 499-507 (1982) and resumed by JC POTTAGE, ibid. 30, (No. 2), pages 215-219, (1986), taking into account the following definitions:

Y _a = infectious titre in the presence of Prp / TV title,

Y _b = infectious titre in the presence of DeG / TV title,

Y _ab = infectious titre in the presence of Prp + DeG / TV title,

Y _c = Y _a XY _b .

Yes :

1) Yab <Y _c , the interaction is a synergy,

2) Y _ab > Y _c niais <to the most active compound alone, the interaction is in this case a subadditive reaction,

3) Y _ab > the least active compound alone, the interaction is an antagonism, and

4) Y _ab > the most active compound but <the least active compound, the interaction is then called interference.

To simplify the calculations, the infectious titres found during the tests of the Deg / Prp associations and the control tests were expressed by their decimal logarithm. The results obtained are recorded in Table I below.

The results in Table I show that:

at the highest concentrations used (Prp 0.363 mg / ml and DeG 50 mg / ml), it is not possible to observe an interaction since at this dose the propolis alone is virucidal,

at the Prp concentration of 0.272 mg / ml, only an interference effect with DeG is observed,

- On the other hand, for low concentrations of Prp (0.090 to 0.180 mg / ml), there is a synergy with DeG occurring at the concentration of 25 or 50 mg / ml.

In the case in point, there is synergy when the ratio Rp = DeG / Prp is between 135/1 and TABLE I: Study of Prp / DeG Association on the Infectious Title of HSViR

TV Title Title Essays Ya Yb Yab Yc Interaction

Prp DeG Prp

+ DEG

4.5 3.5 (a) 2 (f) 0.5 0.77 0.44 0.11 0.33 synergy

4.5 4.5 (a) 3 (g) 2.5 1 0.66 0.55 0.66 synergy

5.0 4.0 (a) 2 (g) 0 0.8 0.4 0 0.32 synergy

4.5 2.5 (b) 2 (f) 0 0.55 0.44 0 0.24 synergy

4.0 1.0 (c) i (f) 0 0.25 0.25 0 0.06 synergy

4.5 1.5 (c) 2 (f) 0 0.33 0.44 0 0.14 synergy

4.5 3.5 (c) 3 (g) 2 0.77 0.66 0.44 0.50 synergy

5.0 3.0 (c) 2 (g) 1 0.6 0.4 0.2 0.24 synergy

4.0 0 (d) 2 (f) 0.5 0 0.5 0.12 0 interference

5.0 1.0 (d) 2 (g) 1.0 0.2 0.4 0.25 0.08 interference

4.0 0 (e) 2 (f) 0 0 0.5 0 0 (h)

Notes

(a) Ppr at the concentration of 0.090 mg / ml

(b) Prp at the concentration of 0.136 mg / ml

(c) Prp at the concentration of 0.180 mg / ml

(d) Prp at the concentration of 0.272 mg / ml

(e) Prp at the concentration of 0.363 mg / ml

(f) DeG at the concentration of 50 mg / ml

(g) DeG at the concentration of 25 mg / ml

(h) dose too high in Prp

EXAMPLE 2 - Formulation -

A mixture of 59 g of white petrolatum, 2.99 g of sorbitan sesquioleate and 3 g of glycerol monooleate is heated to 70-75 ° C. The mixture is ceased to heat when the mixture has become homogeneous and then 5 g of DEG, 30 g of water at a temperature of 65 ° C. or less are added with stirring and the stirring is continued until cooling to RT. Homogenizes and obtains an ointment consisting of a water-in-oil emulsion, used as eye drops.

EXAMPLE 3 - Formulation -

56.98 g of white petrolatum are mixed at 70-75 ° C. with 3.5 g of sorbitan sesquioleate and 3.5 g of glycerol monooleate. When the mixture has become homogeneous, the heating is stopped and 7 g of DeG are added. Mixed again and added with stirring a mixture consisting of 0.02 g of propolis and 30 g of water at a temperature below about 70 ° C. Stirring is continued until cooling to RT, then homogenized and obtained an ointment consisting of a water-in-oil emulsion, used as eye drops. EXAMPLE 4 - Formulation -

4.66 g of polyethylene glycol 1500 stearate are mixed with 13 g of glycerol monostearate, 3 g of glycerol monooleate, 10.5 g of decyl oleate, 5.5 g of capric / caprylic triglyceride and 5 g of glycerol monostearate. glycerol isostearate. The mixture is gradually heated to 70-75 ° C and the heating is stopped as soon as the mixture has become homogeneous. 8 g of DeG are then added and the mixture is stirred slowly. 3 g of propylene glycol, 0.3 g of citral and 47 g of water are then poured into the resulting mixture. The resulting mixture is stirred while allowing it to cool to RT. After passing through a homogenizer, a water-in-oil emulsion having the consistency of a cream is obtained. EXAMPLE 5 - Obtaining the purified propolis -

The propolis used in Examples 1 and 3 above is a purified propolis obtained by extraction with EtOH at 80% [ie EtOH / H ₂ O mixture in a weight ratio of approximately (8/2)], as indicated above. after. The raw Prp is deposited by the bees on perforated plastic grates mounted on frames arranged in the hives. These grids containing the crude Prp are taken from the hives and brought to -30 ° C in a freezer so as to make the crude Prp brittle, which is then grinded with mortar. The milled material is extracted with 80% EtOH, 1 part by weight of crude Prp for 10 parts by volume of 80% EtOH, for 24 hours, with stirring, at RT. The insoluble residue is removed by filtration. The filtrate which is collected is evaporated under reduced pressure and gives a dry extract of light brown color. Yield: 65% by weight, based on the initial crude Prp.

Claims

1. Galvinoxyl derivative chemically named 2,6-di-t-Butyl- (3,5-di-t-butyl-4- 2,5-cyclohexadien-1-ylidene) -p-tolylhydroxy and represented by the following formula :

2. galvinoxyl derivative according to claim 1, for use in the treatment of diseases caused by the strains of herpes HSVI, _HSV2 and / or HSViR.

3. Galvinoxyl derivative according to claim 1 or 2, characterized in that it is associated with propolis.

4. Pharmaceutical composition characterized in that it comprises a galvinoxyl derivative as defined in claim 1.

5. Pharmaceutical composition according to claim 4, characterized in that it comprises propolis.

6. Composition according to claim 5, characterized in that it comprises from 100 to 650 parts by weight of the galvinoxyl derivative for one part by weight of propolis.

A composition according to any one of claims 4 to 6 for use in the treatment of diseases caused by lipoprotein enveloped viruses (LEVs).

8. Composition according to claim 7, for its use in the treatment of diseases caused by the herpes HSVi, HSV ₂ and / or HSViR strains.