EP0433353A1 - Procede et composition de traitement et de prevention d'infections virales - Google Patents

Procede et composition de traitement et de prevention d'infections virales

Info

Publication number
EP0433353A1
EP0433353A1 EP89910134A EP89910134A EP0433353A1 EP 0433353 A1 EP0433353 A1 EP 0433353A1 EP 89910134 A EP89910134 A EP 89910134A EP 89910134 A EP89910134 A EP 89910134A EP 0433353 A1 EP0433353 A1 EP 0433353A1
Authority
EP
European Patent Office
Prior art keywords
virus
protease
infection
viral infection
viral
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.)
Pending
Application number
EP89910134A
Other languages
German (de)
English (en)
Inventor
Peter Morgan Sutton
John Sydney Oxford
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.)
Public Health Laboratory Service Board
Hvivo Services Ltd
Original Assignee
Public Health Laboratory Service Board
Hvivo Services 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 Public Health Laboratory Service Board, Hvivo Services Ltd filed Critical Public Health Laboratory Service Board
Publication of EP0433353A1 publication Critical patent/EP0433353A1/fr
Pending legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/43—Enzymes; Proenzymes; Derivatives thereof
    • A61K38/46—Hydrolases (3)
    • A61K38/48—Hydrolases (3) acting on peptide bonds (3.4)
    • A61K38/4873—Cysteine endopeptidases (3.4.22), e.g. stem bromelain, papain, ficin, cathepsin H
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/16—Antivirals for RNA viruses for influenza or rhinoviruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants

Definitions

  • This invention relates to a method and composition for the treatment and prevention of viral infections.
  • the invention is of particular utility in the treatment and prevention of infections with enveloped viruses including, for example human immuno-deficiency viruses (HIV) and influenza viruses.
  • enveloped viruses including, for example human immuno-deficiency viruses (HIV) and influenza viruses.
  • AIDS Acquired Immune Deficiency Syndrome
  • the causative agent of AIDS is *a retrovirus previously referred to as HTLV-III or LAV, but now referred to as HIV.
  • Current therapies proposed for combatting HIV infections generally involve the use of anti-viral agents which interfere with viral replication, for example by inhibiting reverse transcriptase. Such agents are generally cytotoxic and despite the limited success which has been achieved with such anti-viral agents as zidovudine (AZT) , no effective treatment of AIDS has been found to date.
  • the present invention is based on the discovery that proteolytic. enzymes (proteases) may be used in the therapy of viral infections. This is unexpected in that enzymes play only a minor role as therapeutic drugs, for example streptokinase is used as a X- fibrinolytic agent while asparaginase forms part of most combination therapies for childhood acute lymphatic leukaemia. Certain enzymes have been administered orally in an attempt to reduce soft tissue inflammation, notably the bromelains, which are proteases obtained from pineapple plants. However the British National Formulary emphasises that their therapeutic effectiveness is doubtful. An available dosage form of bromelains has until recently been marketed by Rorer Pharmaceuticals (a subsidiary of Fisons Limited) under the proprietory brand name "Ananase Forte”.
  • Bromelains have further been used in skin care lotions, for promoting healing and have also been proposed as plasminogen activators in the treatment of thrombo-occlusive vascular disease. There has never, however, been any suggestion of the use of bromelains or other proteases as antiviral agents.
  • a method of treating a viral infection in a subject which comprises administering a protease.
  • the invention further provides the use of a protease in the production of a pharmaceutical composition for use in treatment of a viral infection in a subject.
  • virus as used herein is intended to include any entity capable of infecting a eukaryotic cell, whereby the infected cell can produce duplicates of the infectious entity.
  • virus embraces DNA- or RNA-based infectious particles where the nucleic acid is associated with other macromocular species such as, for example, proteins and lipids.
  • prions are infectious entities implicated in such diseases as scrapie in sheep, bovine spongiform encephalopathy in cattle, and Kuru and Creutzfeltd-Jakob disease in man.
  • the method of the invention may be applied to the prevention and treatment of a wide range of viral infections.
  • examples include infections with retroviruses such as HIV referred to previously, and with other enveloped viruses such as, e.g. influenza viruses.
  • enveloped viruses usually have surface glycoprotein "spikes" (peplomers) .
  • a further application of proteases in accordance with the invention comprises the production of vaccines for providing a protective effect against viral infection in humans or animals.
  • a method for the production of inactivated or attenuated virus for incorporation into a vaccine or vaccine component which comprises contacting said virus with a protease.
  • proteases in accordance with the invention include the use of proteases in the treatment of a food product which is contaminated with virus or suspected of having such contamination, which comprises contacting said food product with said protease.
  • food products include meat suspected of being contaminated with viruses or with the causative agents (prions) responsible for scrapie in sheep and bovine spongiform encephalopathy in cattle.
  • proteases may be used in carrying out the method of the invention and in general, the enzyme of choice will be determined by the nature of the viral infection being treated and the required route of administration. Thus, for example, where the nature of the viral infection makes it necessary for the protease to T be introduced into the systemic circulation, the selected protease should have a relatively low immunogenicity and/or toxicity. It is also advantageous in such circumstances to be able to use a protease which is capable of entering the systemic circulation following administration of a suitable dosage form.
  • proteases of non-animal origin are preferably used in accordance with the invention.
  • a plant protease such as, for example, a bromelain, papain or ficin.
  • Processes for producing these proteases are described in US Patent Nos 2950227 and 3002891. The latter specifically relates to the extraction of- bromelains from pineapple stems. However both stem and fruit bromelains may be used in the method of the invention.
  • proteases useful in accordance with the invention include digestive enzymes such as, for example trypsin and chymotrypsin.
  • the protease may be administered by any convenient means.
  • it may be administered orally or parenterally, or in the case of a local viral infection, it may be administered topically.
  • Parenteral modes of administration include administration by injection intravenously, intramuscularly, subcutaneously, intrathecally or into a body cavity, e.g. intraperitoneally or i trapleurally.
  • the protease is administered orally in which case, in order to avoid inactivation in the stomach, the protease is preferably administered in the form of an enteric-coated tablet or pill, or in the form of a capsule resistant to degradation in the stomach.
  • rectal administration in the form of a suppository, in the form of an aerosol as in a nasal spray or as eye drops.
  • a suitable dose is dependent on the mode of application, the toxicity of the selected protease and the proteolytic activity of the protease. For specific combinations of these parameters, effective dosage ranges may be determined by carrying out appropriate in vitvo and in vivo experiments. Generally it has been found that concentrations of proteases in the range 1-100 ⁇ g/ml, particularly 5 ⁇ 100 ⁇ g/ml can exert an anti-viral effect and dosages capable of producing such levels in the blood-stream are advantageously employed.
  • Proteases may be used in accordance with the present invention in combination therapies together with other therapeutic agents having an antiviral activity.
  • combination therapies involving the simultaneous or successive administration of a protease and zidovudine (AZT) are included within the scope of the present invention.
  • a particularly useful combination therapy which does not require the use of zidovudine or related nucleoside analogues comprises administering, simultaneously or successively, a protease in accordance with the present invention and a surfactant and/or a steroid in accordance with the procedures described in our European Patent Application No. 85302275-8 (EP-0285285) .
  • compositions produced in accordance with the invention may comprise conventional pharmaceutically acceptable diluents or carriers.
  • typical injectable solutions will comprise sterile pyrogen-free media, e.g. normal saline and optionally include buffering agents, stabilising agents and preservatives.
  • conventional enteric coatings may be used.
  • Tablets and pills may include conventional excipients such as, for example, lactose, starch and magnesium stearate while suppositories will include excipients such as waxes and ' glycerol.
  • compositions according to the invention are topical compositions for treating and/or protecting a subject against viral infection.
  • Such compositions may comprise a protease and an excipient adapted for topical application.
  • Such compositions may for example be in the form of creams, ointments, lotions, solutions or gels.
  • Topical compositions in forms suitable for vaginal use e.g. creams, ointments, gels, foams and soluble pessaries
  • These compositions may advantageously contain a spermicidally active agent.
  • a protease may be used in the production or sterilisation of pharmaceutical preparations (particularly injectable pharmaceutical preparations) and surgical devices in order to reduce the content of (or contamination with) infectious virus, pro-virus, prion or virus-infected cells therein.
  • pharmaceutical preparation as used herein is meant any substance, composition, living tissue, implantable material or prosthetic device useful in medical treatment or therapy.
  • surgical device is intended to include any surgical apparatus or instrument, or any product or device used in a surgical treatment, including, e.g. a suture or catheter.
  • Examples of pharmaceutical preparations include whole blood (for transfusion), blood plasma or other blood products (e.g. Factor VIII) .
  • the protease may be added so as to produce a concentration sufficient to inactivate any virus present. The concentration should not be so high, or the treatment sufficiently long to cause substantial deterioration of the product being treated.
  • Proteases may be used in accordance with the invention for treating tissues for use as grafts and transplants, in order to reduce the likelihood of transmission of viral infections. This is particularly important in the transplantation of e.g. corneal tissue, or in transplant techniques in which living cells are brought into contact with nerve cells. Thus in the past, individuals have inadvertently been infected with the infectious agent of Creutzfeldt-Jakob disease by transplantation of infected corneal tissue.
  • a further utility of proteases in accordance with the invention comprises applying a protease to a food product in order to eliminate virus particles therefrom.
  • a protease for example, meat products may be treated with proteases in accordance with the invention in order to eliminate contamination with virus. This procedure is particularly useful in eliminating prions from animal- derived foodstuff destined for human or animal consumption.
  • the antiviral activity of bromelain on three strains of influenza virus was assessed according to the following protocol.
  • the bromelain used in this and subsequent experiments was obtained from Siber Hegner Limited and had the following characteristics:
  • Samples of allantoic fluid infected with influenza virus were mixed with varying concentrations of bromelain and incubated at 37°C for 1 hour in phosphate buffered saline (PBS) or in broth saline at pH 7.0.
  • PBS phosphate buffered saline
  • Two strains of influenza A virus (HINI and H3N2) and one strain of influenza B virus (B/Yamagata/10/88) were used in the experiments.
  • the incubated fluids were then titrated for residual virus infection using 10 day-old embryonated hen's eggs by injecting samples into the eggs using standard techniques.
  • the antiviral activity of bromelain on HIV was assessed using the persistently HIV-infected human T-lymphocytes cell line known as "H9" (more specifically the H*9 III B cell line). Uninfected cells (an established human T-cell line, specifically the C8l66 cell line) were used as markers.
  • H9 cells at a concentration of 2 x 10 cells/ml, were incubated in wells of a 2-4 well icrotitre dish (0.5 ml/well) in RPMI
  • RPMI 1640 medium is a complex mixture of amino acids, salts, glucose and vitamins, obtainable from Gibco BRL under Catalogue Ref.
  • Bromelain stock solution was serially diluted to concentrations of 200, 20, 2.0 and 0.2 ⁇ g/ml, and 0.5 ml aliquots were added to the wells to give final concentrations of 100, 10, 1.0 and
  • virus inhibiting effect was particularly significant at a bromelain concentration of between 0.5 ⁇ g/ml and 50 ⁇ g/ .
  • the toxicity of bromelain to CEM cells, C8l66 cells and cord blood•lymphocytes was determined in the following manner for each cell line.
  • a suspension of the cells in RPMI 1640 with hepes and 1-glut + 3% FCS (plus interleuki ⁇ 2 for the cord blood lymphocytes) was centrifuged and the cell pellet resuspended in the same medium at a concentration of 10 cells/ml 0.75 ml aliquots of the suspension were placed in wells of 24-well tissue culture plate and 0.75 ml of bromelain solution added to selected wells.
  • concentrations of bromelain solution were used in the experiment:
  • the two established cell lines CEM and C8166 were somewhat more sensitive to bromelain if cultured for 4 days or longer.
  • protease acts by removing the glycoprotein spikes (peplomers) which are characteristic of enveloped viruses, leaving the resulting bald particles non-infectious.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Public Health (AREA)
  • Chemical & Material Sciences (AREA)
  • Veterinary Medicine (AREA)
  • Virology (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Animal Behavior & Ethology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Communicable Diseases (AREA)
  • Oncology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Engineering & Computer Science (AREA)
  • Pulmonology (AREA)
  • Molecular Biology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Epidemiology (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicinal Preparation (AREA)

Abstract

L'invention concerne un procédé de traitement d'une infection virale chez un sujet, consistant à administrer une enzyme de protéase, notamment une protéase de plante telle que par exemple une broméline, papaïne ou ficine. On peut citer à titre d'exemples d'infections virales pouvant être traitées des infections ayant pour origine des virus contenant de l'ADN ou de l'ARN ou des infections provoquées par des prions. L'invention concerne également l'utilisation de protéases dans la production ou la stérilisation de préparations pharmaceutiques ou de matières alimentaires afin de réduire la teneur en cellules virales, pro-virales ou infectées par un virus dans celles-ci, ainsi que l'utilisation de protéases pour la stérilisation ou la décontamination de dispositifs chirurgicaux. L'invention concerne en outre la production d'un virus inactivé ou atténué destiné à être incorporé dans un vaccin ou un composant de vaccin afin d'assurer un effet protecteur contre les infections virales chez l'homme et l'animal.
EP89910134A 1988-09-08 1989-09-07 Procede et composition de traitement et de prevention d'infections virales Pending EP0433353A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB8821049 1988-09-08
GB888821049A GB8821049D0 (en) 1988-09-08 1988-09-08 Method & composition for treatment & prevention of viral infections

Publications (1)

Publication Number Publication Date
EP0433353A1 true EP0433353A1 (fr) 1991-06-26

Family

ID=10643240

Family Applications (2)

Application Number Title Priority Date Filing Date
EP89309061A Withdrawn EP0358500A1 (fr) 1988-09-08 1989-09-07 Méthode et composition en vue du traitement et de la prévention des infections virales
EP89910134A Pending EP0433353A1 (fr) 1988-09-08 1989-09-07 Procede et composition de traitement et de prevention d'infections virales

Family Applications Before (1)

Application Number Title Priority Date Filing Date
EP89309061A Withdrawn EP0358500A1 (fr) 1988-09-08 1989-09-07 Méthode et composition en vue du traitement et de la prévention des infections virales

Country Status (6)

Country Link
EP (2) EP0358500A1 (fr)
JP (1) JPH04500518A (fr)
AU (1) AU4205789A (fr)
GB (1) GB8821049D0 (fr)
WO (1) WO1990002562A1 (fr)
ZA (1) ZA896864B (fr)

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Also Published As

Publication number Publication date
WO1990002562A1 (fr) 1990-03-22
EP0358500A1 (fr) 1990-03-14
ZA896864B (en) 1990-06-27
JPH04500518A (ja) 1992-01-30
AU4205789A (en) 1990-04-02
GB8821049D0 (en) 1988-10-05

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