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
  • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
  • A61K47/32—Macromolecular compounds obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. carbomers, poly(meth)acrylates, or polyvinyl pyrrolidone
• • 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
  • A61P37/00—Drugs for immunological or allergic disorders
  • A61P37/02—Immunomodulators
  • A61P37/04—Immunostimulants
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P39/00—General protective or antinoxious agents

Definitions

• the present invention relates to the field of vaccines. More particularly, the invention relates to the stabilization of liquid vaccines.
• the invention provides a method for stabilizing a vaccine composition in the liquid state according to which high molecular weight polyvinylpyrrolidone is added to the vaccine composition.
• the subject of the invention is also a liquid vaccine composition stable over time, characterized in that it comprises at least one viral antigen, and at least high molecular weight polyvinylpyrrolidone
• the molecular weight of PNP is greater than or equal to 100,000 daltons.
• the PVP is used at a concentration of at least 0.1% by weight, and preferably less than or equal to 5%.
• the vaccine composition comprises live attenuated viruses. It may especially be a vaccine composition against polio, intended for oral administration.
• the vaccine composition further comprises salts or sugars as well as at least one surfactant. Under these conditions, the stability results obtained are particularly satisfactory.
• the vaccine compositions whose stability over time should be ensured are vaccine compositions comprising at least one antigen constituted by a virus.
• the stability of such compositions is assessed, according to criteria defined by the various regulatory authorities, by maintaining, over time, the infectious titer of the vaccine composition, which means that the viruses used as antigens must retain their capacity to infect cells.
• vaccines comprising live attenuated viruses and, in particular, vaccines against poliomyelitis.
• They can be monovalent vaccine compositions, that is to say intended for protection against a single disease (even if they can comprise several types of the same valence, as is the case with polio or Dengue fever, for example) or multivalent compositions, that is to say vaccines intended for protection against several diseases, at least one of the valences being constituted by a live attenuated virus as described above.
• the method according to the invention has shown all its interest for the stabilization of the oral polio vaccine, which is a live attenuated virus vaccine comprising the 3 types of polio virus.
• the vaccine composition is stabilized thanks to an addition of Polyvinylpyrrolidone (or PVP) with high molecular weight, in particular PVP360 whose PM is 360,000 Daltons.
• PVP Polyvinylpyrrolidone
• high molecular weight PVP makes it possible to very advantageously replace the albumin usually used in the stabilizing formulations of liquid vaccine compositions. The good quality of the results obtained is all the more surprising since it contradicts the teaching of US Pat. No. 3,915,794, according to which the PVP K90 which has a MW of 360,000 is not satisfactory for the stabilization of viral suspension .
• the vaccine composition according to the invention also comprises a certain amount of surfactant, such as Polyethylene glycol (or PEG). Tween TM 80 or Polysorbate 80 can also be used.
• the amount of surfactant used is preferably an amount of less than 0.007% w / v.
• the vaccine composition further comprises salts such as magnesium chloride MgCl 2 , in substantially molar concentration; the vaccine composition can also comprise sugars such as glucose and sucrose, the concentrations of which can vary from approximately 20% to approximately 40%; however, the presence of these sugars is not critical for the stabilizing effect.
• salts such as magnesium chloride MgCl 2
• sugars such as glucose and sucrose, the concentrations of which can vary from approximately 20% to approximately 40%; however, the presence of these sugars is not critical for the stabilizing effect.
• the vaccine composition can also include any other element usually used in vaccines, such as preservatives and / or adjuvants. It can in particular comprise buffer substances, such as the Hepes buffer in a concentration equal to approximately 20 mM.
• Viral suspensions of the 3 different types of polio are produced as follows:
• a biogenerator containing Vero cells at the 142nd passage, in Parker 199 medium, is inoculated with one of the types of polio virus.
• the viral culture is carried out at a temperature of approximately 34 ° C.
• the viral harvest is carried out which is carried out by recovery of the supernatant.
• the harvest is filtered on a membrane having a cutoff threshold of 100,000, then purified by passage through a DEAE Spherodex column previously balanced at pH 7 in 40mM phosphate buffer. Under these conditions, the impurities are retained by the column while the viruses pass freely through it.
• the viral suspension having passed through the column is then filtered on a membrane having a cutoff threshold of 10,000, then subjected to a zonal ultracentrifugation on a sucrose gradient; the fraction of interest is that present at 45-55% sucrose.
• a monovalent suspension of a given type (I, II or III) of polio virus is thus obtained.
• Vaccine compositions are prepared from the monovalent compositions prepared in Example 1 by mixing: type I viral suspension, type II viral suspension, type III viral suspension, in amounts allowing reach a titer for each of the 6.3 log10DICC50 / dose strains under a volume of 100 ml, to which one of the following mixtures is added, in order to obtain the compositions whose stability is to be tested:
• the stability of the viral compositions prepared is evaluated by assessing the decrease in the infectious titer over time.
• the determination of the infectious titer is carried out by the DICC50 technique which is carried out as follows: The titration is carried out on 96-well microplates.
• the samples are diluted from -1 to -6 or -7 (in log10) in MEM lxC medium with 2% (volume / volume) of fetal calf serum not containing antibodies against polio, 4% (volume / volume) of sodium bicarbonate 5.6% (volume / volume) without phenol red and 0.2% of a solution of antibiotic penicillin-didromycin 400xC (volume / volume). For each dilution, distribute 50 ⁇ l in 4 rows of 10 wells:
• antiserum is added 50 .mu.l of polio type2 more polio type 3 antiserum diluted das MEM medium to neutralize polio virus type 2 and 3 contained in the vaccine composition (titration type I), - in the 3 rd row, 50 ⁇ l of typical anti-polio antiserum plus anti-polio type 3 antiserum diluted in d ⁇ MEM medium are added to neutralize the polio type 1 and 3 viruses contained in the vaccine composition (type II titration),
• the 8 wells in the last column of each of the plates serve as cell controls: 100 ⁇ l of MEM medium are placed therein per well.
• the plates are covered by means of a cover; it is stirred sideways and left in contact for 1 hour at 37 ° C. After this contact time, 100 ⁇ l of HepII cell suspension at 50,000 cells / ml are added to each well. The plates are placed for 9 days in an oven at 36 ° C. To prepare the cell suspension, one must proceed as follows: dissociate the cells by the action of trypsin, count and dilute the cell suspension to obtain a concentration of 50,000 cells / ml.
• the pathogenic effects are read in each of the wells after checking the integrity of the cell controls.
• the titer is calculated using a linear regression on the different dilutions after transforming the data into a sine arc of the root of the proportions of positive wells.
• the assays are carried out, for each of the viral compositions prepared, at T0 and after 5 days at 37 ° C.

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• Health & Medical Sciences (AREA)
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• Public Health (AREA)
• General Health & Medical Sciences (AREA)
• Animal Behavior & Ethology (AREA)
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• Chemical Kinetics & Catalysis (AREA)
• Inorganic Chemistry (AREA)
• Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
• General Chemical & Material Sciences (AREA)
• Organic Chemistry (AREA)
• Immunology (AREA)
• Epidemiology (AREA)
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• Virology (AREA)
• Communicable Diseases (AREA)
• Engineering & Computer Science (AREA)
• Bioinformatics & Cheminformatics (AREA)
• Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
• Medicinal Preparation (AREA)

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• 2000
  • 2000-10-06 FR FR0012805A patent/FR2814957B1/fr not_active Expired - Fee Related
• 2001
  • 2001-10-08 KR KR1020037004927A patent/KR100832551B1/ko not_active IP Right Cessation
  • 2001-10-08 MX MXPA03002875A patent/MXPA03002875A/es active IP Right Grant
  • 2001-10-08 WO PCT/FR2001/003097 patent/WO2002028362A2/fr active IP Right Grant
  • 2001-10-08 CA CA002424863A patent/CA2424863A1/en not_active Abandoned
  • 2001-10-08 AU AU9567201A patent/AU9567201A/xx active Pending
  • 2001-10-08 JP JP2002531988A patent/JP2004526667A/ja active Pending
  • 2001-10-08 BR BR0114350-6A patent/BR0114350A/pt not_active IP Right Cessation
  • 2001-10-08 NZ NZ525117A patent/NZ525117A/en unknown
  • 2001-10-08 EP EP01976379A patent/EP1357895A2/fr not_active Withdrawn
  • 2001-10-08 CN CNB018169708A patent/CN1298387C/zh not_active Expired - Fee Related
  • 2001-10-08 AU AU2001295672A patent/AU2001295672B2/en not_active Ceased
  • 2001-10-08 US US10/381,948 patent/US6982088B2/en not_active Expired - Fee Related

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