GB2355192A - Anti-viral treatment - Google Patents

Anti-viral treatment Download PDF

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Publication number
GB2355192A
GB2355192A GB9924389A GB9924389A GB2355192A GB 2355192 A GB2355192 A GB 2355192A GB 9924389 A GB9924389 A GB 9924389A GB 9924389 A GB9924389 A GB 9924389A GB 2355192 A GB2355192 A GB 2355192A
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GB
United Kingdom
Prior art keywords
lithium
virus
effect
treatment
anti
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
GB9924389A
Other versions
GB9924389D0 (en
Inventor
Ian Pardo
Christopher Hartley
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Henderson Morley Research and Development Ltd
Original Assignee
Henderson Morley Research and Development Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Henderson Morley Research and Development Ltd filed Critical Henderson Morley Research and Development Ltd
Priority to GB9924389A priority Critical patent/GB2355192A/en
Publication of GB9924389D0 publication Critical patent/GB9924389D0/en
Priority claimed from GB0000683A external-priority patent/GB0000683D0/en
Publication of GB2355192A publication Critical patent/GB2355192A/en
Withdrawn legal-status Critical Current

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Classifications

    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/70Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • A61K31/704Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K33/00Medicinal preparations containing inorganic active ingredients

Abstract

A composition for use against viral infections comprising a cardiac glycoside and a lithium salt. The cardiac glycosides include digoxin, digitoxin, medigoxin, lanatoside C, proscillaridin, k strophanthin, peruvoside and ouabain. The composition is particularly effective against DNA viruses such as Herpes Simplex Virus (HSV) and Cytomegalovirus (CMV). In a preferred embodiment, viral infections of the eye are treated by the application of contact lenses impregnated with the compostion.

Description

2355192 ANTI-VIRAL TREATMENT The invention relates to anti-viral

treatments and in particular to prophylactic and therapeutic treatment of Herpes infections.

The invention is based on the discovery that the use of a cardiac glycoside and lithium salts has a particularly enhanced effect on viral infections.

According to the invention in one aspect there is provided a therapeutic composition useful in the treatment of viral infections comprising a cardiac glycoside and lithium.

In another aspect the invention provides a method of treating a viral infection comprising the application of a cardiac glycoside and lithium.

The cardiac glycosides may be any one or more of digoxin, digitoxin, medigoxin, lanatoside C, proscillaridin, k strophanthin, peruvoside and ouabain. Plants of the digitalis species (e.g. digitalis purpura, digitalis lanata) contain cardiac glycosides such as digoxin and digitoxin which are known collectively as digitalis. Other plants contain cardiac glycosides which are chemically related to the digitalis glycosides and these are often also referred to as digitalis.

At the molecular level digitalis exerts it main effect by the inhibition of the sodium transport enzyme sodium potassium adenosine triphosphate (Na/K ATPase); this is directly responsible for the electrophysiological effects of heart muscle and also 2 its anti-viral activity. This activity also has an effect on the efficiency of myocardial contractility due to secondary changes 'in intracellular calcium. At very low intracellular concentrations of digitalis, the opposite effects can be seen with a reduced efficiency of cardiac contractions as the digitalis stimulates the Na/K ATPase.

The lithium is preferably provided by a water soluble or water miscible salt or ester: preferred are lithium chloride, lithium carbonate and lithium sulphate.

Lithium is known to affect the behaviour and replication of bacteria and viruses. Hadley et al (1931) reported that incorporation of lithium chloride (12mM) into beef infusion broth encouraged the generation of filterable G forms of the Shiga bacillus and Morishita and Tokada (1976) reported that while lithium had a sodium-sparing effect for the growth of Vibrio parahaemolyticus, higher concentrations, in excess of 300mM, inhibited growth of the organism. In 1979, Leighton found inhibition of growth of a number of Gram negative bacteria by comparatively low concentrations of lithium chloride (12mM), and recently we found that higher concentrations (80mM) inhibit growth of Gram positive as well as Gram negative bacteria (Buchan et al. 1989).

Lithium specifically inhibits replication of the deoxyribonucleic acid viruses, HSV, pseudorabies and vaccinia but not replication of the ribonucleic acid viruses, influenza and encephalomyocarditis (Skinner et al 1980). The effects of lithium on macromolecular synthesis in virus infected cells has not been extensively studied. Ziaie and Kefalides (1989) indicated that HSV induced suppression of host cell polypeptide synthesis was less marked when cells were cultured in the presence of 3 lithium (20 -30mM) and that there was synthesis of virus specific polypeptides under those conditions.

Lithium influx will inevitably alter the intracellular concentration of other cations and it is believed that inhibition of HSV replication by lithium is a direct consequence of that alteration. Potassium, for example, has been implicated in the control of macro-molecular synthesis; in BHK cells, decreased intracellular potassium inhibited protein synthesis, DNA synthesis and cell replication (McDonald et al 1972).

The substitution of potassium by lithium and the inhibition of the NaATPase pump both have an effect that is independent of each other, however our invention discloses that the effect of simultaneous administration of lithium and cardiac glycosides is much greater than each administered separately.

DNA viruses have a central core of DNA. The DNA carries the molecular code to produce the virus proteins, however the function of many viral proteins is unknown. There are however several that are well characterised including important biological catalysts (enzymes) which act as ideal targets for chemotherapy. These include DNA polymerase which is needed for replication, a protease enzyme which cleaves proteins, and an important enzyme called thymidine kinase.

When a vir-us takes over a host cells biosynthesis machinery the replication of the DNA code is fundamental to virus reproduction.

4 Acyclovir (Zovirax Wellcome) was the first ever genuinely selective anti- viral medication and belongs to the class of drugs known as DNA polymerase inhibitors or nucleoside analogues This has certainly made a massive impact into the treatment of life- threatening herpes infections (e.g. herpes encephalitis), and a significant impact for many in the treatment of the primary attack and recurrences of herpetic disease.

The drugs exert their action in two stages. Firstly the drug will become altered (phosphorylated) by the enzyme thymidine kinase (from the virus). Secondly the triphosphate form of the drug then acts as an 'inhibitory substance for viral DNA polymerase (the enzyme which enable chains of DNA to be formed within the host cell). This prevents new infectious virus particles from being made. Because thymidine kinase is not found in normal or uninfected cells, the action of acyclovir is restricted to virus infected cells only. This gives the drug a high degree of selectivity and as a consequence means that its side effect profile is quite good.

According to this invention, treatment of infection caused by DNA viruses by cardiac glycosides and lithium may be of benefit against several classes of virus.

The compositions of the invention may be adapted for external or internal administration using known carriers. Topical and systemic applications are likely to be most useful. Other ingredients may be present, provided that they do not Dist compromise the anti-viral activity.

Our investigations have shown the synergistic effect of the cardiac glycoside and lithium is present in the proportions of 25 mcg/ml Onabain and 20 mM lithium (as a water soluble salt). Our evaluations suggest that the proportions of the two ingredients may vary.

Although our investigations suggest that the synergistic effect takes place with the simultaneous application of the cardiac glycoside and the lithium, it is within the scope of the invention to separate the applications by a short time period.

A preferred embodiment of this invention is the use of high local concentrations of lithium and ouabain for the treatment of virus infections of the eye.

Recurrent herpes simplex infection of the cornea in man is the largest cause of blindness due to virus infections on a global basis.

The application of contact lenses impregnated with lithium and cardiac glycosides would be a safe and efficient method for creating high intracellular concentrations and thus prevent and treat the disease.

A depot application of lithium and cardiac glycosides applied intraoccularly would be a suitable method for the treatment of Cytornegalovirus retinitis, a major cause of blindness in patients suffering with AIDS.

In order that the invention may be well understood it will now be described by way of illustration only with reference to the following examples.

Example 1 In vitro assay - 6 Bioassays with herpes simplex virus in vitro were undertaken to follow the antiviral activity of the simultaneous administration of digoxin and lithium.

Culture and assay methods follow those described in Lennette and Schmidt (1979) for herpes simplex virus and Vero cells with minor modifications.

Herpes simplex strains used - Type 1 herpes simplex strain HFEM is a derivative of the Rockerfeller strain HF (Wildy 1955) and Type 2 herpes simplex strain 3345 a penile isolate (Skinner et al 1977) were used as prototype strains. These prototypes were stored at -80deg until needed.

Cell cultures African Green Monkey kidney cells (Vero) were obtained from the National Institute of Biological Standards and Control UK and were used as the only cell line for all experiments in this Example.

Culture Media Cells and viruses were maintained on Glasgows modified medium supplemented with 10% foetal bovine serum.

This example used 20 mM lithium (as a soluble salt solution) and 25 mcg of Ouabain as a stock solution.

The effects of this stock solution on Vero cells in culture was examined, as were the effects of Vero cells which had been infected with type 1 and type 2 virus at varying levels of infectivity. The effect of the stock solution on extracellular virus was also examined.

7 Results The stock solution was found to be non toxic to cell monolayers and as much as 10% of the glycoside could be recovered from the medium after 72 hours incubation.

The effects of the stock solution on different Multiplicities of Infection (i.e. concentration of virus) may be summarised as below:

Inhibition of hsv2 cylovathic effect by ouabain solution alone Multiplicity of infection (Dose of Effect of ouabain solution alone virus) High Medium + Low ++ + signifies minor inhibition of virus cytopathic effect, - signifies no inhibition of virus cytopathic effect.

Inhibition of hsv2 cytopathic effect by lithium solution alone Multiplicity of infection (Dose of Effect of lithium solution alone virus) High Medium + Low ++ 8 + signifies minor inhibition of virus cytopathic effect, - signifies no inhibition of virus cytopathic effect.

Inhibition of hsv2 cytopathic effect ... signifies inhibition of virus cytopathic effect i.e. the action of the virus on destroying the vero cells was prevented.

This example demonstrates that virus activity was prevented by applying a small volume of stock solution to Vero cells infected with virus and that the effects of this stock solution was far greater that the effects of lithium or ouabain applied by themselves. Further experiments were completed to examine effects on extracellular virus. There was no direct virucidal activity on extracellular virus.

Other evaluations have shown that at lower concentration of stock solution this anti-viral activity disappears in a concentration dependent manner.

Multiplicity of infection (Dose of Effect of lithium/ glycoside solution virus) High....

Medium....

Low....

9 ExaMRIe 2 Example I was repeated to determine the effect on Herpes Simplex typel and other herpes viruses and almost identical results were obtained.

Claims (4)

1. Use of a cardiac glycoside and a lithium salt in the treatment of viral infections.
2. A therapeutic composition useful for the treatment of virus infections in subjects comprising an effective antiviral amount of the composition 'in claim 1 and a suitable carrier.
3. A therapeutic composition in accordance with claim 2 adapted for topical application.
4. A therapeutic composition in accordance with claim 2 adapted for systemic application.
GB9924389A 1999-10-15 1999-10-15 Anti-viral treatment Withdrawn GB2355192A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
GB9924389A GB2355192A (en) 1999-10-15 1999-10-15 Anti-viral treatment

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
GB9924389A GB2355192A (en) 1999-10-15 1999-10-15 Anti-viral treatment
GBGB9930811.6A GB9930811D0 (en) 1999-10-15 1999-12-30 Anti-viral treatment
GBGB9930813.2A GB9930813D0 (en) 1999-10-15 1999-12-30 Anti-viral treatment
GB0000683A GB0000683D0 (en) 1999-10-15 2000-01-12 Antiviral treatment

Publications (2)

Publication Number Publication Date
GB9924389D0 GB9924389D0 (en) 1999-12-15
GB2355192A true GB2355192A (en) 2001-04-18

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GB9924389A Withdrawn GB2355192A (en) 1999-10-15 1999-10-15 Anti-viral treatment
GBGB9930811.6A Ceased GB9930811D0 (en) 1999-10-15 1999-12-30 Anti-viral treatment
GBGB9930813.2A Ceased GB9930813D0 (en) 1999-10-15 1999-12-30 Anti-viral treatment

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GBGB9930811.6A Ceased GB9930811D0 (en) 1999-10-15 1999-12-30 Anti-viral treatment
GBGB9930813.2A Ceased GB9930813D0 (en) 1999-10-15 1999-12-30 Anti-viral treatment

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007026124A2 (en) * 2005-09-02 2007-03-08 Henderson Morley Plc Transdermal delivery means of a diuretic agent and/or cardiac glycoside agent
WO2007026125A2 (en) * 2005-09-02 2007-03-08 Henderson Morley Plc Anti-viral topical gel formulations containing a diuretic such as furosemide and/or a cardiac glycoside such as digoxin
WO2015089494A1 (en) * 2013-12-13 2015-06-18 Ankenman Ralph Compositions and methods for treating dysregulated systems

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4886670A (en) * 1983-07-27 1989-12-12 Efamol Limited Anti-viral compositions
EP0391410A1 (en) * 1989-04-06 1990-10-10 Daiichi Pharmaceutical Co., Ltd. Antiviral composition
EP0442744A2 (en) * 1990-02-14 1991-08-21 TSUMURA & CO. Treatment of viral disease by glycosides

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4886670A (en) * 1983-07-27 1989-12-12 Efamol Limited Anti-viral compositions
EP0391410A1 (en) * 1989-04-06 1990-10-10 Daiichi Pharmaceutical Co., Ltd. Antiviral composition
EP0442744A2 (en) * 1990-02-14 1991-08-21 TSUMURA & CO. Treatment of viral disease by glycosides

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2007026124A2 (en) * 2005-09-02 2007-03-08 Henderson Morley Plc Transdermal delivery means of a diuretic agent and/or cardiac glycoside agent
WO2007026125A2 (en) * 2005-09-02 2007-03-08 Henderson Morley Plc Anti-viral topical gel formulations containing a diuretic such as furosemide and/or a cardiac glycoside such as digoxin
WO2007026125A3 (en) * 2005-09-02 2007-09-27 Christopher Hartley Anti-viral topical gel formulations containing a diuretic such as furosemide and/or a cardiac glycoside such as digoxin
WO2007026124A3 (en) * 2005-09-02 2008-09-25 Christopher Hartley Transdermal delivery means of a diuretic agent and/or cardiac glycoside agent
WO2015089494A1 (en) * 2013-12-13 2015-06-18 Ankenman Ralph Compositions and methods for treating dysregulated systems

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Publication number Publication date
GB9930811D0 (en) 2000-02-16
GB9924389D0 (en) 1999-12-15
GB9930813D0 (en) 2000-02-16

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