EP0857069A1 - Procedes de preservation de micro-organismes - Google Patents

Procedes de preservation de micro-organismes

Info

Publication number
EP0857069A1
EP0857069A1 EP96935088A EP96935088A EP0857069A1 EP 0857069 A1 EP0857069 A1 EP 0857069A1 EP 96935088 A EP96935088 A EP 96935088A EP 96935088 A EP96935088 A EP 96935088A EP 0857069 A1 EP0857069 A1 EP 0857069A1
Authority
EP
European Patent Office
Prior art keywords
micro
amphiphile
organisms
virus particles
polyoxyethylene
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Withdrawn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
EP96935088A
Other languages
German (de)
English (en)
Inventor
Roger Randal Charles New
Charles Anthony The University of Liverpool HART
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.)
Cortecs Ltd
Original Assignee
Cortecs 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 Cortecs Ltd filed Critical Cortecs Ltd
Publication of EP0857069A1 publication Critical patent/EP0857069A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/44—Oils, fats or waxes according to two or more groups of A61K47/02-A61K47/42; Natural or modified natural oils, fats or waxes, e.g. castor oil, polyethoxylated castor oil, montan wax, lignite, shellac, rosin, beeswax or lanolin
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N7/00—Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • A61K39/125—Picornaviridae, e.g. calicivirus
    • A61K39/13—Poliovirus
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
    • A61K47/12—Carboxylic acids; Salts or anhydrides thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A61K9/107—Emulsions ; Emulsion preconcentrates; Micelles
    • A61K9/1075—Microemulsions or submicron emulsions; Preconcentrates or solids thereof; Micelles, e.g. made of phospholipids or block copolymers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
    • A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
    • 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
    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
    • A61K47/24—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing atoms other than carbon, hydrogen, oxygen, halogen, nitrogen or sulfur, e.g. cyclomethicone or phospholipids
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/32011—Picornaviridae
    • C12N2770/32051—Methods of production or purification of viral material
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
    • C12N2770/00011—Details
    • C12N2770/32011—Picornaviridae
    • C12N2770/32611—Poliovirus
    • C12N2770/32634—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the present invention relates to methods of preserving micro-organisms such that they retain their infectivity.
  • the invention relates to methods of preserving viral particles.
  • Vaccines comprising viral particles have been in use for a number of years. It is, however, essential that such vaccines can be stored, sometimes for long periods, without the viral component losing its infectivity.
  • Common storage methods include freezing or freeze-drying, the latter usually involving reconstitution using water at a later stage.
  • certain viruses display reduced viability/infectivity when subjected to these processes .
  • One virus which is not suitably stored as described above is polio virus. This virus is readily degraded at room temperature in aqueous suspension, is stable for only two weeks at 0°C and is destroyed by lyophilisation.
  • preferred methods of storage involve freezing at -70°C or refridgeration at 4°C.
  • such storage conditions are not particularly suitable for use in tropical countries or indeed countries where the required facilities and equipment are scarce.
  • compositions comprising a hydrophilic species solubilised in a hydrophobic phase, as well as methods for their preparation.
  • UK application no. 9424901.8 discloses compositions as - described in PCT/GB94/02495 which incorporate additional components which aid retention of the hydrophilic species in the hydrophobic phase.
  • UK application no.9424902.6 discloses compositions as described in PCT/GB94/02495 which incorporate moieties which aid formation of the composition.
  • UK patent application no. 9422990.3 discloses immunogenic compositions which comprise an immunogen solubilised, suspended or otherwise dispersed in a hydrophobic phase.
  • the immunogen can be a virus and the compositions are useful as vaccines .
  • micro-organisms particularly virus particles, such as polio virus particles
  • virus particles such as polio virus particles
  • polio virus particles may be converted to a form suitable for long term storage at ambient temperature, with retention of infectivity after reconstitution in aqueous medium.
  • such compositions have particular advantages for use in countries where the ususal storage methods are less appropriate, and provide an effective means by which such viruses can be transported and stored without the need for extreme freezing or prolonged refridgeration.
  • the present invention provides a method of storing micro-organisms such that they maintain infectivity, which method includes the steps of :-
  • micro-organisms are virus particles particularly polio virus particles.
  • Suitable methods for carrying out the above method are those described in PCT/GB94/02495, UK 9424901.8, UK 9424902.6 and UK 9422990.3.
  • the hydrophobic solvent could for example be a long chain fatty acid, a medium chain alcohol, a branched long chain alcohol, a monoglyceride, a diglyceride, a medium chain triglyceride, a long chain triglyceride, a halogenated (e.g. fluorinated) analogue thereof, or a polyoxyethylene-containing lipid.
  • the hydrophobic solvent is a mono-, di- or tri-glyceride, or oleic acid.
  • the method comprises : (i) co-dispersing the micro-organisms with an amphiphile in a liquid medium;
  • the liquid medium can be water, and it can be removed by, e.g. freeze drying, centrifugal vacuum drying or any other suitable method.
  • the amphiphile will be a phospholipid, for instance one with a phosphatidyl choline head group, eg phosphatidyl choline (PC) , lysophosphatidyl choline (lyso-PC) , sphingomyelin or a derivative of one of these such as hexadecyl phosphocholine or an amphiphile polymer containing phosphoryl choline.
  • a phosphatidyl choline head group eg phosphatidyl choline (PC) , lysophosphatidyl choline (lyso-PC) , sphingomyelin or a derivative of one of these such as hexadecyl phosphocholine or an amphiphile polymer containing phosphoryl choline.
  • a phosphatidyl choline head group eg phosphatidyl choline (PC) , lysophosphati
  • a bile salt a glycolipid, a polyoxyethylene containing surfactant, a lipophilic sulphate, betaine, a sarcosine containing surfactant, Solulan C24 , polyoxyethylene 40 stearate, one of the Tween series of surfactants, one of the Span series of surfactants or a pegolated castor oil derivative, e.g. Cremaphor EL35.
  • the micro ⁇ organisms eg virus particles
  • This array is then in turn coated with the hydrophobic solvent .
  • access to the micro-organisms by water is restricted, which in turn accounts for the improved storage properties when the micro-organism preparation is reconstituted from a freeze-dried state.
  • the present invention provides a method of storing micro-organisms such that they retain infectivity, which method includes the following steps:
  • the water is removed by freeze-drying.
  • the amphiphile can be a phospholipid, for instance one with a phosphatidyl choline head group, eg phosphatidyl choline (PC) , lysophosphatidyl choline (lyso-PC) , sphingomyelin or a derivative of one of these such as hexadecyl phosphocholine or an amphiphile polymer containing phosphoryl choline.
  • a phosphatidyl choline head group eg phosphatidyl choline (PC) , lysophosphatidyl choline (lyso-PC) , sphingomyelin or a derivative of one of these such as hexadecyl phosphocholine or an amphiphile polymer containing phosphoryl choline.
  • a phosphatidyl choline head group eg phosphatidyl choline (PC) , lysophosphati
  • a bile salt a glycolipid, a polyoxyethylene containing surfactant, a lipophilic sulphate, betaine, a sarcosine containing surfactant, Solulan C24, polyoxyethylene 40 stearate, one of the Tween series of surfactants, one of the Span series of surfactants or a pegolated castor oil derivative, e.g. Cremaphor EL35.
  • the amphiphile is Solulan C24, polyoxyethylene 40 stearate, one of the Tween series of surfactants, one of the Span series of surfactants or a pegolated castor oil derivative, e.g Cremaphor EL35.
  • the amphiphile is Solulan C24 or polyoxyethylene 40 stearate.
  • the method also includes the step of elevating the temperature of the mixture after removal of the water. This ensures that the structure adopted by the amphiphile/micro-organism array . is more condensed, which in turn results in more restricted access for water upon reconstitution.
  • the amphiphile will be one which remains solid after the water removal step, eg it can be chosen from a phospholipid, for instance lecithin, a glycolipid, a polyoxyethylene containing surfactant, a lipophilic sulphate, betaine, a sarcosine containing surfactant, Solulan C24 , polyoxyethylene 40 stearate, one of the Tween series of surfactants, one of the Span series of surfactants or a pegolated castor oil derivative, e.g. Cremaphor EL35.
  • a phospholipid for instance lecithin
  • a glycolipid e.g., a polyoxyethylene containing surfactant, a lipophilic sulphate, betaine, a sarcosine containing surfactant, Solulan C24 , polyoxyethylene 40 stearate, one of the Tween series of surfactants, one of the Span series of surfactants or a pegolated castor oil derivative,
  • a micro-organism composition obtainable by any of the methods described herein, particularly a micro- organism composition comprising virus particles, eg polio virus particles; and
  • composition of the invention for the storage of virus particles.
  • Preferred features of each aspect of the invention are as for each other aspect mutatis mutandis .
  • a suspension of IO 9 polio virus particles (Sabin strains, Types 1, 2, 3) per ml of culture was diluted 1000-fold in distilled water. 1ml of the diluted suspension was mixed with 1ml of adispersion of sonicated soya phospholipid
  • control vial (at a concentration of lOOmg/ml) in distilled water.
  • a control vial was prepared which contained virus only, without the addition of phospholipid.
  • a control vial of polio virus was prepared as above. To this control vial, containing virus alone, was added lml of culture medium.
  • lO ⁇ l of oil/virus preparation was transferred to a fresh vial, and lml of a 2% solution of ox bile extract (containing predominantly sodium taurocholate) was added.
  • Example 2 A virus suspension (Sabin strains, Types 1, 2, 3) containing 5xlO e particles/ml (spun to remove contaminating protein) was diluted 50-fold by addition of 200 ⁇ l of the suspension to 9.9ml of distilled water, yielding a concentration of IO 7 particles/ml. The suspension was divided into four equal aliquots of 2.5ml, and dispensed into 7ml screw-capped glass vials. One aliquot was employed in the experiment described herein, while two were used in the experiment described in example 3.
  • sonicated phospholipid dispersion (lOOmg/ml) was added to the aliquot of diluted virus particles with gentle mixing. 200 ⁇ l of this mixture was dispensed into 20 freeze-drying vials, and the remainder was transferred, In lOO ⁇ l aliquots, into other tubes as "pre ⁇ drying" controls. The controls were stored overnight at +4°C. The freeze-drying vials were placed in the centrifugal rotor of the freeze-dryer and lyophilised overnight .
  • the suspensions prepared above were used to perform 10- fold dilutions in Vero cell monolayer cultures, in order to measure the viability of the polio virus present, the results were expressed as the highest dilution at which 50% cytopathic effects were observed. Nature of sample Highest dilution at which 50% cytopathic effects were observed. Nature of sample Highest dilution at which 50% cytopathic effects were observed. Nature of sample Highest dilution at which
  • the suspensions prepared as described herein were used to perform 10-fold dilutions in Vero cell cultures, to measure the viability of the polio virus present. The results were expressed as the highest dilution at which 50% cytopathic effects were observed.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Public Health (AREA)
  • Veterinary Medicine (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Virology (AREA)
  • Epidemiology (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Microbiology (AREA)
  • Oncology (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Communicable Diseases (AREA)
  • Mycology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Dispersion Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

On décrit des procédés permettant de préserver des micro-organismes, y compris des virus, de façon à maintenir leur infectivité, et on décrit l'utilisation de tels procédés pour préparer des vaccins par exemple.
EP96935088A 1995-10-25 1996-10-25 Procedes de preservation de micro-organismes Withdrawn EP0857069A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
GBGB9521806.1A GB9521806D0 (en) 1995-10-25 1995-10-25 Preservation methods
GB9521806 1995-10-25
PCT/GB1996/002615 WO1997015331A1 (fr) 1995-10-25 1996-10-25 Procedes de preservation de micro-organismes

Publications (1)

Publication Number Publication Date
EP0857069A1 true EP0857069A1 (fr) 1998-08-12

Family

ID=10782855

Family Applications (1)

Application Number Title Priority Date Filing Date
EP96935088A Withdrawn EP0857069A1 (fr) 1995-10-25 1996-10-25 Procedes de preservation de micro-organismes

Country Status (12)

Country Link
EP (1) EP0857069A1 (fr)
JP (1) JP2000501282A (fr)
KR (1) KR19990067029A (fr)
CN (1) CN1202829A (fr)
AU (1) AU714485B2 (fr)
BR (1) BR9610932A (fr)
CA (1) CA2235495A1 (fr)
GB (1) GB9521806D0 (fr)
NO (1) NO981865L (fr)
NZ (1) NZ320446A (fr)
WO (1) WO1997015331A1 (fr)
ZA (1) ZA969015B (fr)

Families Citing this family (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7645608B2 (en) 2004-08-17 2010-01-12 Pml Microbiologicals, Inc. Microorganism specimen storage, hydrating, transfer and applicator device
ES2708989T3 (es) 2010-03-31 2019-04-12 Stabilitech Biopharma Ltd Método de conservación de adyuvantes de alumbre y vacunas potenciadas con alumbre
DK2552478T3 (en) * 2010-03-31 2017-03-27 Stabilitech Ltd EXCIPIENTS FOR STABILIZING VIRUS PARTICLES
GB2499479A (en) 2010-03-31 2013-08-21 Stabilitech Ltd Stabilisation of viral particles
KR102023207B1 (ko) 2010-12-02 2019-11-25 온콜리틱스 바이오테크 인코포레이티드 동결건조 바이러스 제형
AU2011336413B2 (en) 2010-12-02 2015-01-22 Oncolytics Biotech Inc. Liquid viral formulations
GB201117233D0 (en) 2011-10-05 2011-11-16 Stabilitech Ltd Stabilisation of polypeptides
GB201406569D0 (en) 2014-04-11 2014-05-28 Stabilitech Ltd Vaccine compositions
GB2562241B (en) 2017-05-08 2022-04-06 Stabilitech Biopharma Ltd Vaccine compositions
WO2022020249A1 (fr) 2020-07-20 2022-01-27 Stratix Labs Corporation Dispositifs et procédés d'inoculation d'une cible

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2158935A1 (fr) * 1993-10-12 1995-04-20 Chiron Viagene, Inc. Procedes de conservation de virus de recombinaison
GB9323588D0 (en) * 1993-11-16 1994-01-05 Cortecs Ltd Hydrophobic preparation
GB9424901D0 (en) * 1994-12-09 1995-02-08 Cortecs Ltd Sequestration Agents
GB9424902D0 (en) * 1994-12-09 1995-02-08 Cortecs Ltd Solubilisation Aids

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9715331A1 *

Also Published As

Publication number Publication date
NO981865L (no) 1998-06-24
ZA969015B (en) 1998-04-28
BR9610932A (pt) 1999-12-21
NO981865D0 (no) 1998-04-24
CA2235495A1 (fr) 1997-05-01
KR19990067029A (ko) 1999-08-16
GB9521806D0 (en) 1996-01-03
AU714485B2 (en) 2000-01-06
WO1997015331A1 (fr) 1997-05-01
NZ320446A (en) 1999-05-28
CN1202829A (zh) 1998-12-23
JP2000501282A (ja) 2000-02-08
AU7318396A (en) 1997-05-15

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