WO2008117089A2

WO2008117089A2 - Treatments for viral infections

Info

Publication number: WO2008117089A2
Application number: PCT/GB2008/050213
Authority: WO
Inventors: Richard George Stead
Original assignee: Richard George Stead
Priority date: 2007-03-23
Filing date: 2008-03-25
Publication date: 2008-10-02
Also published as: GB0705557D0; WO2008117091A3; EP2124971A2; WO2008117091A2; CA2682145A1; WO2008117089A3

Abstract

A treatment for a viral infection, the treatment comprising the products formed from a peroxidase catalysed reaction between an oxidisable species and at least one oxygen donor, in aqueous solution; wherein the original peroxidase enzyme has been removed therefrom.

Description

A Treatment

The present invention relates to a treatment, a method of preparing the same and a kit for preparing the same.

Viral infections are caused by a foreign species (virus) colonizing in a host organism and utilizing the hosts resources in order to multiply. Viral infections can have severe detrimental effects in the host including inflammation of the infected area which can lead in some cases to severe trauma to the host and even death. Common viral infections that may affect humans include: chickenpox; common cold; measles; herpes simplex; and human immunodeficiency virus (HIV) .

Commonly, anti-viral infection agents are used to treat a viral infection by killing or preventing the growth of the virus without harming the host. Many different antiviral infection agents have been developed and are effective against different viruses, but can have a detrimental effect on the host.

It is an aim of objects of the present invention to provide an alternative anti-viral infection agent that does not have a detrimental effect on the host and a treatment for one or more viral infection.

According to a first aspect of the present invention there is provided a treatment for a viral infection, the treatment comprising the products formed from a peroxidase catalysed reaction between an oxidisable species and at least one oxygen donor, in aqueous solution; wherein the original peroxidase enzyme has been removed therefrom. The treatment may be orally administrable, topically administrable, administrable as a wash or douche, or administrable by inhalation. The treatment may be orally administrable as an aqueous solution.

The treatment may be administered topically or orally any of a spray, a gel, a bath, a wetted cloth.

The treatment may be administered by inhalation by spray, humidified air or fog.

For the avoidance of doubt, the term "treatment" as used herein refers to a pharmaceutical composition.

Preferably, the oxidisable species comprises any species operable to be oxidised by the oxygen donor in the presence of a peroxidase enzyme.

In one embodiment, the oxidisable species may comprise a source of thiocyanate ions. The source of thiocyanate ions may be any substance that provides free thiocyanate ions in aqueous solution, for example a metal salt thereof such as any of a sodium, potassium, lithium salt etc. Alternatively, the source of thiocyanate ions may be derived from hydrogen thiocyanate. In one embodiment, the source of thiocyanate ions may be as reaction products of a further reaction.

For the avoidance of doubt, reference herein to thiocyanate ions means [SCN]^" ions.

In an alternative embodiment, the oxidisable species may comprise a source of halide ions. The source of halide ions may be any substance that provides free halide ions in aqueous solution, for example a metal halide salt. The source of halide ions may comprise a source of one or more of fluoride ions, chloride ions, bromide ions or iodide ions. Suitable metal salts include any halide salt that readily dissociates in aqueous solution to provide the free halide ion. For example, the salt may be a group I metal salt such as lithium, sodium, potassium etc .

Preferably, the treatment further comprises an amount of the oxidisable species. For example, the treatment may comprises the products formed from a peroxidase catalysed reaction between an oxidisable species and at least one oxygen donor, in aqueous solution; and an amount of unreacted oxidisable species. Preferably, the oxidisable species is present in the treatment in an amount (by mol) in excess of the said formed products. Preferably, the ratio of the oxidisable species present in the treatment to the amount of the said formed products is at least about 1.1:1 (by mol), more preferably at least about 1.2:1 (by mol), more preferably at least about 1.3:1 (by mol), most preferably at least about 1.4:1. In a particularly preferred embodiment, the ratio of the oxidisable species present in the treatment to the amount of the said formed products is about 1.5:1 (by mol) .

In one embodiment, the oxidisable species may be [SCN]^" and the said formed products may comprise [OSCN]^". In such a scenario, [SCN]^" is present in the treatment in an amount (by mol) in excess of the [OSCN]^".

Alternatively, the oxidisable species present in the treatment may be different to the oxidisable species from which the said formed products are derived. For example, the oxidisable species from which the said products are formed may be [SCN]^", whereas the oxidisable species present in the treatment may be a source of halide ions, such as Cl^", for example.

The source of the oxidisable species may be present in the reactants (prior to the peroxidase catalysed reaction) in a concentration such that the oxidisable species in the reactants is present in the aqueous solution at a concentration of about 0.1 to 5OmM. More preferably, about 0.5 to 1OmM, more preferably between about 1 and 5mM. In a most preferred embodiment, about 2mM.

Preferably, the at least one oxygen donor comprises a source of peroxide.

The source of peroxide may be any entity that comprises the group X-O-O-Y, where 0 represents an oxygen radical. In one embodiment, one of or both of X and Y may be hydrogen. For example, the source of peroxide may be hydrogen peroxide.

Either of X and/or Y may represent a chemical species, such as an organic fragment or, in one embodiment, may represent a surface to which the -0-0- group is bound. If X and/or Y represents a surface to which the -0-0- group is bound, then a linking group between the -0-0- group and the surface may be present, such as a bivalent organic bridging group, for example. Alternatively, X and/or Y may represent an inorganic species, such as magnesium, sodium etc. The source of peroxide may be present in the reactants

(prior to the peroxidase catalysed reaction) in a concentration such that the peroxide species in the reactants in aqueous solution is present at a concentration of about 0.1 to 5OmM. More preferably, about 0.5 to 1OmM, more preferably between about 1 and 5mM.

In one embodiment, the source of peroxide may be an organic peroxide compound or salt thereof.

Preferably, the peroxidase enzyme is a lactoperoxidase enzyme .

The lactoperoxidase may be naturally occurring lactoperoxidase, for example as extracted from milk. Alternatively, the lactoperoxidase may be artificially synthesised.

Preferably, the peroxidase enzyme is provided on one or more support. Preferably, the support is an inorganic clay material. For example, the support may be bentonite.

Preferably, the peroxidase enzyme is present in the reactants (prior to the peroxidase catalysed reaction) in an amount of between about 0.001 to 10 g/1. More preferably, 0.005 to 5 g/1.

Preferably, the peroxidase enzyme is present in the treatment in an amount less than about 0.001 g/1. More preferably, the peroxidase enzyme is present in the treatment in an amount less than about 0.0005g/l. More preferably, the peroxidase enzyme is present in the treatment in an amount less than about O.OOOlg/1. More preferably, the peroxidase enzyme is present in the treatment in an amount less than about O.OOOOlg/1. Preferably, the treatment comprises substantially no peroxidase enzyme therein.

The reaction can occur over a range of pH depending on the desired use of the resulting active species. For example, the reaction can occur between pH 3 and pH 11 (measured at 25⁰C) . In a preferred embodiment, the reaction takes place between pH 4 and pH 10 (measured at 25⁰C) .

Preferably, the reaction is performed in a temperature range between 2⁰C and 6O⁰C. However, it should be noted that the stability of the oxidised products is temperature dependent and therefore a temperature may be chosen to correspond with the intended use. For example, at higher temperatures, the stability of the oxidised species is lower (and therefore degrades faster) than at lower temperatures.

Preferably, the reaction products (the oxidised species) are present in the aqueous solution in a concentration between about 1 to lOOOppm, more preferably between about 5 to 500 ppm, more preferably, between about 10 to lOOppm and most preferably between about 25 and 90ppm. In a most preferred embodiment, the reaction products (the oxidised species) are present in the aqueous solution in a concentration of between about 55 to 70ppm.

The reaction is quite rapid and the products liable to degrade, thus after contacting the reactants at room temperature, it is preferred that the resultant treatment is administered within about 3 to 4 hours. In a preferred embodiment, the peroxidase enzyme is removed from the reaction products formed from the peroxidase catalysed reaction by rendering it captive in a solid phase after the reaction has occurred. For example, the enzyme may be rendered captive in a solid phase by an aggregate, which may be a coagulant, flocculant or emulsifier, which may be in the presence of one or more thickening agent.

Reference herein to "the peroxidase enzyme" should be understood as optionally also referring to the support upon which the enzyme may be carried.

The aggregate may be synthetic or naturally occurring. The aggregate may be Aluminium based. The aggregate may comprise chitosan, which may be derived from an extract of the exoskeleton of crustaceans such as shrimps or crabs for example.

Preferably, the enzyme is rendered captive in a solid phase by one or more flocculant. Preferably, the one or more flocculant is selected from one or more of polymeric anionic or cationic flocculants, such as polysaccharides or polyacrylamides .

Preferably, the enzyme is rendered captive in a solid phase by one or more coagulants.

It is believed that the reaction products of the reaction are the oxidised species (the product of oxidisation of the oxidisable species). Therefore, for example, if the oxidisable species is a thiocyanate ion, then the main active ingredient of the treatment is believed to be hypothiocyanate : [OSCN]^".

According to a further aspect of the present invention, there is provided a method of preparing a treatment, the method comprising contacting at least one oxidisable species with at least one oxygen donor in the presence of a peroxidase enzyme and thereafter removing the peroxidase enzyme.

Preferably, the peroxidase enzyme is removed by one or more removal steps, which may comprise contacting the enzyme with one or more removal agents. The removal agent may comprise one or more aggregate, which may be a coagulant, flocculant or emulsifier.

According to a further aspect of the present invention there is provided a kit for preparing a treatment, the kit comprising a source of an oxidisable species, a source of at least one oxygen donor and a peroxidase enzyme .

The kit may further comprise at least one flocculating agent .

The kit may further comprise a pH adjusting agent such as an acid, for example. The pH adjusting agent may adjust the pH of the aqueous solution to between 3 and 11.

According to a further aspect of the present invention there is provided a use of the reaction products of a source of an oxidisable species and a source of an oxygen donor in the presence of a peroxidase enzyme, the reaction products having been separated from the peroxidase enzyme, in the treatment of a viral infection.

According to a further aspect of the present invention there is provided a pharmaceutical composition comprising an active ingredient and one or more inactive ingredient, the active ingredient consisting of the products formed from a peroxidase catalysed reaction between an oxidisable species and at least one oxygen donor.

The one or more inactive ingredient may comprise a solvent .

The active ingredient may consist of one or more of the following ions: [OSCN]^"; [OF]^"; [OCl]^"; [OBr]^"; [0I]^".

Examples of viral infections and viral infectious diseases include but are not restricted to the following: AIDS; AIDS Related Complex; HIV; Chickenpox (Varicella) ; Common cold; Cytomegalovirus Infection; Colorado tick fever; Dengue fever; Ebola haemorrhagic fever; Hand, foot and mouth disease; Hepatitis; Herpes simplex; Herpes zoster; HPV; Influenza (Flu); Lassa fever; Measles; Marburg haemorrhagic fever; Infectious mononucleosis; Mumps; Poliomyelitis; Progressive multifocal leukencephalopathy; Rabies; Rubella; SARS; Smallpox (Variola); Viral encephalitis; Viral gastroenteritis; Viral meningitis; Viral pneumonia; West Nile disease; Yellow fever.

Preferably, the infection is a respiratory system infection. Preferably, the treatment is for mammalian infections, more preferably, human infections. Preferably, the treatment is a respiratory system infection treatment. In a particularly preferred embodiment, the infection is HIV.

According to a further aspect of the present invention there is provided a use of the reaction products of a source of an oxidisable species and a source of an oxygen donor in the presence of a peroxidase enzyme for the manufacture of a medicament for the treatment of an infection .

The treatment may be administered by adding the aqueous reaction products to a volume of air or an air stream, for example by humidification of air.

Therefore, according to a further aspect of the present invention there is provided a method of treatment of an infection comprising contaminating a volume of air or an air stream with the reaction products of a source of an oxidisable species and a source of an oxygen donor in the presence of a peroxidase enzyme, the peroxidase enzyme having been removed from the said reaction products, and exposing a patient to the said volume of air or air stream.

According to a further aspect of the present invention there is provided a use of the reaction products of a source of an oxidisable species and a source of an oxygen donor in the presence of a peroxidase enzyme, the peroxidase enzyme having been removed from the said reaction products, in destroying airborne infection causing organisms. In a further embodiment of the present invention there is provided the use of the reaction products of a source of an oxidisable species and a source of an oxygen donor in the presence of a peroxidase enzyme, the peroxidase enzyme having been removed from the said reaction products, in contaminating a volume of air or an air stream.

Preferably, the said reaction products are present as an aqueous mist in the volume of air or the air stream. Preferably, the particle diameter of the mist droplets are less than 20 microns in size, more preferably less than 10 microns in size, more preferably between about 1 micron and 5 microns in size. Reference to size of vapour droplets refers to average particle diameter.

Preferably, the aqueous mist is prepared by ultrasound.

All of the features disclosed herein may be combined with any of the above aspects and in any combination.

For a better understanding of the invention, and to show how embodiments of the same may be carried into effect, reference will now be made, by way of example only, to the following experimental data.

A preparation of OSCN- solution was produced by exposing a source of thiocyanate and a source of H₂O₂ to a lactoperoxidase enzyme on a bentonite carrier, followed by adding a flocculant, aluminium polychlorosulphate, to flocculate the enzyme and carrier, before filtering the resulting aqueous solution.

Determination of OSCN^" ions in the reaction medium: The measure is performed with a spectrophotometry technique at 412nm.

OSCN^" content is determined in the reaction medium just before inserting the bacterial inoculum, in order to know precisely the concentration of active principle, which the microorganism population will be exposed to.

A treatment made in accordance with the present invention offers a quick and simple to prepare medicament that may be used to treat a wide range of infections.

Advantageously, a kit may be provided from which the treatment may be manufactured in the home or where required. Such a kit has a long life span and is easily transportable .

An example of such a kit is as follows:

Lactoperoxidase enzyme (on a bentonite support) 1. Og Coagulant (Aluminium polychlorosulphate) 0.0011

Sodium thiocyanate

0.062g

Hydrogen peroxide 0.00751 of

100%

Such a kit may be used to produce a 500ml aqueous solution of 35ppm OSCN^" as follows.

500ml of water is added to a jug followed by adding the enzyme thereto and stirring. The Sodium thiocyanate is then added during continuous stirring. Hydrogen peroxide is next added during continuous stirring before adding the coagulant and stirring for a further 5 minutes. The mixture is then left for about 30 minutes, during which time flocculation occurs and the solid settles to the bottom of the container. The aqueous solution is then poured into a separate receptacle through filter paper and is ready to be used as a treatment, for example by drinking .

Although the above preparation prepares a aqueous solution of 35ppm [OSCN]-, it will be appreciated that by changing the concentration of reagents a different concentration may be prepared, in accordance with the preferred ranges stated above.

Attention is directed to all papers and documents which are filed concurrently with or previous to this specification in connection with this application and which are open to public inspection with this specification, and the contents of all such papers and documents are incorporated herein by reference.

All of the features disclosed in this specification (including any accompanying claims, abstract and drawings), and/or all of the steps of any method or process so disclosed, may be combined in any combination, except combinations where at least some of such features and/or steps are mutually exclusive.

Each feature disclosed in this specification (including any accompanying claims, abstract and drawings) may be replaced by alternative features serving the same, equivalent or similar purpose, unless expressly stated otherwise. Thus, unless expressly stated otherwise, each feature disclosed is one example only of a generic series of equivalent or similar features.

The invention is not restricted to the details of the foregoing embodiment (s) . The invention extends to any novel one, or any novel combination, of the features disclosed in this specification (including any accompanying claims, abstract and drawings), or to any novel one, or any novel combination, of the steps of any method or process so disclosed.

Claims

1. A treatment for a viral infection, the treatment comprising the products formed from a peroxidase catalysed reaction between an oxidisable species and at least one oxygen donor, in aqueous solution; wherein the original peroxidase enzyme has been removed therefrom.

2. A treatment according to claim 1 wherein the treatment is orally administrable, topically administrable, administrable as a wash or douche, or administrable by inhalation.

3. A treatment according to either of claim 1 or claim 2, wherein the treatment is orally administrative as an aqueous solution.

4. A treatment according to any preceding claim, wherein the treatment is administered topically by any of a spray, a gel, a bath, a wetted cloth.

5. A treatment according to any preceding claim, wherein the treatment may be administered by inhalation by spray, humidified air or fog.

6. A treatment according to any preceding claim wherein the oxidisable species comprises any species operable to be oxidised by the oxygen donor in the presence of a peroxidase enzyme.

7. A treatment according to any preceding claim wherein the oxidisable species comprises a source of thiocyanate ions.

8. A treatment according to claim 7, wherein the source of thiocyanate ions comprises any substance that provides free thiocyanate ions in aqueous solution, for example a metal salt thereof such as any of a sodium, potassium, lithium salt etc.

9. A treatment according to claim 7, wherein the source of thiocyanate ions is derived from hydrogen thiocyanate .

10. A treatment according to claim 7, wherein the source of thiocyanate ions is provided as reaction products of a further reaction.

11. A treatment according to claim 1 to 6 wherein the oxidisable species comprises a source of halide ions .

12. A treatment according to claim 11 wherein the source of halide ions is any substance that provides free halide ions in aqueous solution, for example a metal halide salt.

13. A treatment according to claim 11 or 12 wherein the source of halide ions comprises a source of one or more of fluoride ions, chloride ions, bromide ions or iodide ions.

14. A treatment according to claim 11 to 13 wherein suitable metal salts include any halide salt that readily dissociates in aqueous solution to provide the free halide ion.

15. A treatment according to claim 11 to 14 wherein the salt is a group I metal salt such as lithium, sodium, potassium etc.

16. A treatment according to any preceding claim wherein the source of the oxidisable species is present in the reactants (prior to the peroxidase catalysed reaction) in a concentration such that the oxidisable species in the reactants is present in the aqueous solution at a concentration of about 0.1 to 5OmM.

17. A treatment according to any preceding claim wherein the at least one oxygen donor comprises a source of peroxide.

18. A treatment according to claim 17, wherein the source of peroxide is any entity that comprises the group X-O-O-Y, where 0 represents an oxygen radical .

19. A treatment according to claim 18, wherein one of or both of X and Y are hydrogen.

20. A treatment according to claim 18 wherein either of X and/or Y represent a chemical species, such as an organic fragment or, in one embodiment, represent a surface to which the -0-0- group is bound.

21. A treatment according to claim 18 wherein X and/or Y represent an inorganic species, such as magnesium, sodium etc.

22. A treatment according to claim 17 to 21 wherein the source of peroxide is present in the reactants (prior to the peroxidase catalysed reaction) in a concentration such that the peroxide species in the reactants in aqueous solution is present at a concentration of about 0.1 to 5OmM.

23. A treatment according to claim 17 to 22 wherein the source of peroxide is an organic peroxide compound or salt thereof.

24. A treatment according to any preceding claim, wherein the peroxidase enzyme is a lactoperoxidase enzyme .

25. A treatment according to claim 24, wherein lactoperoxidase is naturally occurring lactoperoxidase, for example as extracted from milk .

26. A treatment according to claim 24 wherein the lactoperoxidase has been synthesised artificially.

27. A treatment according to any preceding claim wherein the peroxidase enzyme is provided on one or more support .

28. A treatment according to claim 27, wherein the support is an inorganic clay material.

29. A treatment according to any preceding claim, wherein the peroxidase enzyme is present in the reactants (prior to the peroxidase catalysed reaction) in an amount of between about 0.001 to 10 g/i

30. A treatment according to any preceding claim, wherein the peroxidase catalysed reaction occurs between pH 3 and pH 11 (measured at 25⁰C) .

31. A treatment according to any preceding claim, wherein the peroxidase catalysed reaction is performed in a temperature range between 2⁰C and 6O⁰C.

32. A treatment according to any preceding claim wherein the peroxidase catalysed reaction products (the oxidised species) are present in the aqueous solution in a concentration between about 1 to lOOOppm.

33. A treatment according to any preceding claim wherein the enzyme is removed from the treatment following the peroxidase catalysed reaction by rendering the enzyme captive in a solid phase.

34. A treatment according to claim 33, wherein the enzyme is rendered captive in a solid phase by an aggregate, which may be a coagulant, flocculant or emulsifier, which may be in the presence of one or more thickening agent.

35. A method of preparing a treatment for a viral infection, the method comprising contacting at least one oxidisable species with at least one oxygen donor in the presence of a peroxidase enzyme and thereafter removing the peroxidase enzyme.

36. A kit for preparing a viral infection treatment, the kit comprising a source of an oxidisable species, a source of at least one oxygen donor and a peroxidase.

37. A kit according to claim 36 wherein the kit further comprise at least one flocculating agent.

38. A kit according to claim 36 or 37, wherein the kit further comprises a pH adjusting agent such as an acid, for example.

39. Use of the reaction products of a source of an oxidisable species and a source of an oxygen donor in the presence of a peroxidase enzyme; the peroxidase enzyme having been removed from the products, in the treatment of a viral infection.

40. A treatment according to any of claims 1 to 34 wherein the treatment is for mammalian infections, more preferably, human infections.

41. Use of the reaction products of a source of an oxidisable species and a source of an oxygen donor in the presence of a peroxidase enzyme, the peroxidase enzyme having been removed from the products, for the manufacture of a medicament for the treatment of a viral infection.

42. A method of treatment of a viral infection comprising contaminating a volume of air or an air stream with the reaction products of a source of an oxidisable species and a source of an oxygen donor in the presence of a peroxidase enzyme, the peroxidase enzyme having been removed from the products, and exposing a patient to that said volume of air or air stream.

43. Use of the reaction products of a source of an oxidisable species and a source of an oxygen donor in the presence of a peroxidase enzyme, the peroxidase enzyme having been removed from the products, in destroying airborne viral infection causing organisms.

44. Use of the reaction products of a source of an oxidisable species and a source of an oxygen donor in the presence of a peroxidase enzyme, the peroxidase enzyme having been removed from the products, in contaminating a volume of air or an air stream.

45. A treatment, substantially as herein before described and with reference to the examples.