WO1995003826A1 - Stabilised immunoglobulin preparations and method for preparing same - Google Patents

Stabilised immunoglobulin preparations and method for preparing same Download PDF

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Publication number
WO1995003826A1
WO1995003826A1 PCT/FR1994/000955 FR9400955W WO9503826A1 WO 1995003826 A1 WO1995003826 A1 WO 1995003826A1 FR 9400955 W FR9400955 W FR 9400955W WO 9503826 A1 WO9503826 A1 WO 9503826A1
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concentration
preparations
polyoxyethylene
nonionic surfactant
preparations according
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PCT/FR1994/000955
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French (fr)
Inventor
Michel Gaston Joseph Grandgeorge
Paule Annic Gattel
Marie-France Marguerite Andrée MAKULA
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Pasteur Merieux Sérums Et Vaccins
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Priority to EP94923758A priority Critical patent/EP0711176A1/en
Priority to JP7505621A priority patent/JPH09500894A/en
Priority to AU73866/94A priority patent/AU7386694A/en
Publication of WO1995003826A1 publication Critical patent/WO1995003826A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to preparations' d 1 immunoglobulin (Ig) human or animal, in particular polyclonal immunoglobulins blood.
  • the present invention also relates to a method for stabilizing immunoglobulins.
  • Immunoglobulins are widely used in prophylaxis and therapy. Their mode of administration by intravenous route requires minimizing their anticomplementary activity which can result from denaturation of the Ig leading in particular to the formation of aggregates and polymers.
  • French patent application FR-A-2 301 266 proposes a process for the preparation of injectable gamma globulin intravenously.
  • the pharmaceutical form includes the dissolution of gamma- globulins in a buffered aqueous solution containing glycine and albumin.
  • this application proposes to add to the pharmaceutical preparation obtained a nonionic surfactant.
  • a nonionic surfactant As a surfactant, this application proposes Tweens or Pluronic 68. In the only embodiment described, this application recommends the use of Tween 80 at a concentration of 0.1%, that is to say say 1 g / 1. Consequently, the putting into gamma-globulin pharmaceutical form would pass through the combined use of glycine, albumin and non-ionic surfactant at a concentration of the order of l g / 1.
  • European patent application EP-A 448 075 describes a process for preparing an intravenous IG with membrane filtration, in which it is sought to prevent denaturation of the Ig during filtration using a surfactant stabilizer, which can be a nonionic surfactant such as Pluronic.
  • the recommended stabilizer level is between 0.5 and 50 g / 1.
  • the final product contains the surfactant at a very high rate.
  • patent US-A-4,439,421 recommends not using the nonionic surfactants as described in particular in patent US-A-4,093,606 corresponding to the French patent application above, for a problem. harmless to blood cells.
  • the aforementioned US patent proposes to stabilize the Ig solutions with a view to their lyophilization by combining several types of stabilizers, macromolecules, proteins and low molecular weight polyols. Polyethylene glycol is preferred in conjunction with human albumin and glucose.
  • albumin acts as an effective stabilizer for immunoglobulins. But in the current medical context, we try to avoid as much as possible the use of substances of human or animal origin, which present a risk of viral contamination.
  • the Applicant has now surprisingly discovered that it is possible to prepare stabilized immunoglobulin solutions in liquid form using a non-ionic surfactant in very low concentration, while dispensing with the usual stabilizers such as albumin.
  • the present invention therefore relates to preparations of human or animal immunoglobulins, in particular polyclonal, and in particular polyclonal IgG, which comprise, as preservation stabilizer in liquid form, a nonionic surfactant in concentration less than or equal to 0.1 g / 1 and which are essentially free of albumin.
  • essentially devoid of albumin it should be understood that no trace of albumin is detected by the Pharmeuropa reference method in electrophoresis on cellulose acetate, which corresponds to an amount of albumin- less than 1% of IgG .
  • the concentration of nonionic surfactant is between 0.02 and 0.05 g / l approximately and is preferably of the order of 0.025 g / l.
  • the preparations according to the invention comprise between 30 and 120 g / l of immunoglobulins.
  • the nonionic surfactant is preferably chosen from the group consisting of polyoxyethylene sorbitan monooleate (20), deoaethylene glycol octylphenyl ether, polyoxyethylene sorbitan monolaurate (20), mixed polyoxyethylene / polyoxypropylene and polyoxyethylene copolymer and polyethylene laurate glycol 600.
  • the preparations according to the invention can also comprise a usual lyophilization stabilizer such as sucrose, in particular in a concentration of the order of 50 to 100 g / l.
  • a usual lyophilization stabilizer such as sucrose, in particular in a concentration of the order of 50 to 100 g / l.
  • the immunoglobulin preparations according to the invention also preferably have the characteristics recommended by the standards in force, such as pH between 4.0 and 7.4, osmolality greater than 280 mosmol / kg by the addition of osmotically active solutes , such as mineral salts (such as NaCl) or sugars (such as glucose, sucrose, maltose) or sugar alcohols (such as mannitol, sorbitol) or amino acids (such as glycine).
  • osmotically active solutes such as mineral salts (such as NaCl) or sugars (such as glucose, sucrose, maltose) or sugar alcohols (such as mannitol, sorbitol) or amino acids (such as glycine).
  • the preparations obtained meet the safety criteria for use specific to Ig solutions for intravenous administration, namely bacterial and fungal sterility, apyrogenicity, reduced rate of aggregates and polymers and reduced anticomplementary activity.
  • the immunoglobulins present in the preparation can be obtained from plasma, serum or placenta by conventional methods of protein fractionation, supplemented if necessary by specific treatments aimed at reducing the level of aggregates and polymers and / or to reduce the anticomplementary activity of the preparation (for example moderate treatment with pepsin or with plasmin, dissociative treatment at acid pH, chemical modification by reduction and / or alkylation or precipitation of the aggregates and polymers by PEG).
  • specific treatments aimed at reducing the level of aggregates and polymers and / or to reduce the anticomplementary activity of the preparation (for example moderate treatment with pepsin or with plasmin, dissociative treatment at acid pH, chemical modification by reduction and / or alkylation or precipitation of the aggregates and polymers by PEG).
  • the preparations according to the invention can be ready-to-use solutions, therefore stored in liquid form, or can be solutions obtained extemporaneously by dissolving a lyophilized concentrate.
  • the present invention also relates to a process for stabilizing polyclonal human or animal immunoglobulin preparations, in which a preservation stabilizer in liquid form which is a nonionic surfactant is added to the preparation so as to obtain a concentration less than or equal to 0.1 g / 1 in the final preparation.
  • a preservation stabilizer in liquid form which is a nonionic surfactant is added to the preparation so as to obtain a concentration less than or equal to 0.1 g / 1 in the final preparation.
  • the nonionic surfactant is added in a concentration of between 0.02 and 0.05 g / l approximately and preferably of the order of 0.025 g / l.
  • the nonionic surfactant is chosen from those indicated above.
  • the surfactant is advantageously added just before the final packaging in vials, but it can also be added at an earlier stage in the process of extraction and purification of Ig.
  • Severe conditions for handling intravenous Ig vials were simulated using a shake test.
  • 50 ml type I borosilicate glass vials are aseptically filled with 20 ml of a sterile Ig solution at 50 g / l.
  • the bottles are shaken at room temperature of 20 ° C for 1 or 2 hours on an oscillating / alternating type agitator set for 80 horizontal oscillations per minute.
  • the anticomplementary activity (AcA) of the solution is measured according to the test described in "Pharmeuropa” (supra).
  • Variable concentration of Tween 80 from 0 to 100 mg / 1
  • Triton X 100 (mg / 1) 0 25 50 100 * AcA before shaking 0.37 0.36 0.36 0.31
  • Albumin exerts a protective effect on Ig from a concentration between 100 and 1000 mg / 1.
  • Preferred nonionic surfactants are:
  • Triton X 100 polyoxyethylene octylphenyl ether manufactured by Rohm and Haas.
  • Pluronic F 68 polyoxyethylene and polyoxypropylene copolymer manufactured by Ugine Kuhlmann.
  • Polyethylene glycol 600 laurate (manufactured by Gattefossé).
  • Tween 80 is preferred because of its absence of toxicity and its use in pharmaceutical or food formulation well documented (See the work "Non Ionic Surfactants” M. Schick ed. Marcel Dekker NY, 1967, 28, 923-970) .

Abstract

Substantially albumin-free human or animal and particularly polyclonal immunoglobulin preparations including a non-ionic surfactant in a concentration no higher than 0.1 g/l as a liquid preservative stabilising agent.

Description

Préparations d'immunoglobulines stabilisées et procédé pour leur préparation Stabilized immunoglobulin preparations and process for their preparation
La présente invention a trait à des préparations' d1immunoglobulines (Ig) humaines ou animales, en particulier immunoglobulines polyclonales sanguines. La présente invention a également trait à un procédé de stabilisation des immunoglobulines. Les immunoglobulines sont largement utilisées dans la prophylaxie et la thérapeutique. Leur mode d'administration par voie intraveineuse nécessite de réduire au maximum leur activité anticomplémentaire qui peut résulter d'une dénaturation des Ig conduisant notamment à la formation d'agrégats et de polymères.The present invention relates to preparations' d 1 immunoglobulin (Ig) human or animal, in particular polyclonal immunoglobulins blood. The present invention also relates to a method for stabilizing immunoglobulins. Immunoglobulins are widely used in prophylaxis and therapy. Their mode of administration by intravenous route requires minimizing their anticomplementary activity which can result from denaturation of the Ig leading in particular to the formation of aggregates and polymers.
Ainsi, la norme européenne publiée dans Pharmeuropa (3 (4) , décembre 1991, 259-268) exige que les solutions d1immunoglobulines pour administration intraveineuse aient des taux d'agrégats et de polymères inférieurs ou égaux à 3 % des protéines totales et une activité anticomplémentaire inférieure ou égale à 1 unité CH 50 par g d'Ig. La formation d'agrégats n'intervient pas seulement au cours de la préparation des solutions d'immunoglobulines, mais aussi au cours de leur conservation sous forme liquide, notamment lors de leurs manipulations. En effet, les immunoglobulines ont tendance à se dénaturer aux interfaces liquide/gaz et liquide/solide, ce qui se traduit par une augmentation de l'activité anticomplémentaire par formation d'agrégats" solubles ou insolubles.Thus, the European standard published in Pharmeuropa (3 (4), December 1991, 259-268) requires that solutions of 1 immunoglobulin for intravenous administration have rates of aggregates and polymers or less equal to 3% of the total protein and an anticomplementary activity less than or equal to 1 CH 50 unit per g of Ig. The formation of aggregates occurs not only during the preparation of immunoglobulin solutions, but also during their storage in liquid form, in particular during their handling. Indeed, immunoglobulins tend to denature at the liquid / gas and liquid / solid interfaces, which results in an increase in anticomplementary activity by the formation of " soluble or insoluble aggregates.
La demande de brevet français FR-A-2 301 266 propose un procédé de préparation de gamma-globulines injectables par voie intraveineuse. La mise sous forme pharmaceutique comprend la dissolution des gamma- globulines dans une solution aqueuse tamponnée et contenant du glycocolle et de l'albumine.French patent application FR-A-2 301 266 proposes a process for the preparation of injectable gamma globulin intravenously. The pharmaceutical form includes the dissolution of gamma- globulins in a buffered aqueous solution containing glycine and albumin.
En outre, pour empêcher ou réduire toute denaturation de surface (aux interfaces liquide/air ou liquide/solide), cette demande propose d'ajouter à la préparation pharmaceutique obtenue un agent tensio-actif non ionique. A titre d'agent tensio-actif, cette demande propose les Tweens ou le Pluronic 68. Dans le seul mode de réalisation décrit, cette demande préconise l'utilisation de Tween 80 à une concentration de 0,1 % c'est-à-dire de 1 g/1. En conséquence, la mise sous forme pharmaceutique des gamma-globulines passerait par l'utilisation combinée de glycocolle, d'albumine et de tensio-actif non-ionique à une concentration de l'ordre de l g/1.In addition, to prevent or reduce any surface denaturation (at the liquid / air or liquid / solid interfaces), this application proposes to add to the pharmaceutical preparation obtained a nonionic surfactant. As a surfactant, this application proposes Tweens or Pluronic 68. In the only embodiment described, this application recommends the use of Tween 80 at a concentration of 0.1%, that is to say say 1 g / 1. Consequently, the putting into gamma-globulin pharmaceutical form would pass through the combined use of glycine, albumin and non-ionic surfactant at a concentration of the order of l g / 1.
La demande de brevet européen EP-A 448 075 décrit un procédé de préparation d'ig intraveineuse avec filtration sur membrane, dans lequel on cherche à empêcher la denaturation des Ig lors de la filtration à l'aide d'un stabilisant tensio-actif, qui peut être un tensio-actif non ionique tel que le Pluronic. Le taux de stabilisant recommandé est compris entre 0,5 et 50 g/1. Il en résulte que le produit final contient le tensio- actif à un taux très élevé. H.L. Levine et al. (J. of Parenteral Science andEuropean patent application EP-A 448 075 describes a process for preparing an intravenous IG with membrane filtration, in which it is sought to prevent denaturation of the Ig during filtration using a surfactant stabilizer, which can be a nonionic surfactant such as Pluronic. The recommended stabilizer level is between 0.5 and 50 g / 1. As a result, the final product contains the surfactant at a very high rate. H.L. Levine et al. (J. of Parenteral Science and
Technology, volume 45 (3) mai-juin 1991, pages 160 à 165) rapportent une étude sur les effets protecteurs des tensio-actifs non ioniques contre la denaturation de surface de solutions d'anticorps monoclonaux faiblement concentrées dans un test d'agitation. Les auteurs rapportent une concentration efficace de l'ordre de 0,1 %, c'est-à-dire correspondant à 1 g/1. Aucune protection n'est par contre obtenue avec une concentration de 0,1 g/1. Or il est établi qu'à ces concentrations élevées, ces agents tensio-actifs présentent des inconvénients sérieux en cas d'injection intraveineuse.Technology, volume 45 (3) May-June 1991, pages 160 to 165) report a study on the protective effects of nonionic surfactants against surface denaturation of weakly concentrated monoclonal antibody solutions in a shaking test. The authors report an effective concentration of the order of 0.1%, that is to say corresponding to 1 g / 1. No protection is however obtained with a concentration of 0.1 g / 1. However, it is established that at these high concentrations, these surfactants have serious drawbacks in the event of intravenous injection.
Ainsi, le brevet US-A-4 439 421 recommande de ne pas utiliser les tensio-actifs non ioniques tels que décrits notamment dans le brevet US-A-4 093 606 correspondant à la demande de brevet français ci-dessus, pour un problème d'innocuité vis-à-vis des cellules sanguines. Cela confirme les observations de J.C. KRANTZ et al. (J. Pharmacol Exp. Ther. 93, pages 188 à 195, 1948) sur le pouvoir hémolytique du Tween 20 à la concentration de 1 g/1. Le brevet US précité propose de stabiliser les solutions d'Ig en vue de leur lyophilisation en combinant plusieurs types de stabilisants, macromolécules, protéines et polyols de bas poids moléculaire. Le polyéthylene glycol est préféré en liaison avec de l'albumine humaine et du glucose.Thus, patent US-A-4,439,421 recommends not using the nonionic surfactants as described in particular in patent US-A-4,093,606 corresponding to the French patent application above, for a problem. harmless to blood cells. This confirms the observations of J.C. KRANTZ et al. (J. Pharmacol Exp. Ther. 93, pages 188 to 195, 1948) on the hemolytic power of Tween 20 at the concentration of 1 g / 1. The aforementioned US patent proposes to stabilize the Ig solutions with a view to their lyophilization by combining several types of stabilizers, macromolecules, proteins and low molecular weight polyols. Polyethylene glycol is preferred in conjunction with human albumin and glucose.
Il est en effet connu que 1'albumine agit comme un stabilisant efficace pour les immunoglobulines. Mais dans le contexte médical actuel, on cherche à éviter autant que faire se peut l'utilisation de substances d'origine humaine ou animale, qui présentent un risque de contamination virale.It is indeed known that albumin acts as an effective stabilizer for immunoglobulins. But in the current medical context, we try to avoid as much as possible the use of substances of human or animal origin, which present a risk of viral contamination.
La demanderesse a maintenant découvert de façon surprenante qu'il était possible de préparer des solutions d'immunoglobulines stabilisées sous forme liquide à l'aide d'un tensio-actif non ionique en très faible concentration, tout en se passant des stabilisants habituels tels que l'albumine. La présente invention a donc pour objet des préparations d'immunoglobulines humaines ou animales, en particulier polyclonales, et notamment IgG polyclonales, qui comprennent, à titre de stabilisant de conservation sous forme liquide, un tensio-actif non ionique en concentration inférieure ou égale à 0,1 g/1 et qui sont essentiellement dépourvues d'albumine. Par essentiellement dépourvues d'albumine, il faut comprendre qu'aucune trace d'albumine n'est détectée par la méthode de référence Pharmeuropa en électrophorèse sur acétate de cellulose , ce qui correspond à une quantité d'albumine- inférieure à 1% des IgG.The Applicant has now surprisingly discovered that it is possible to prepare stabilized immunoglobulin solutions in liquid form using a non-ionic surfactant in very low concentration, while dispensing with the usual stabilizers such as albumin. The present invention therefore relates to preparations of human or animal immunoglobulins, in particular polyclonal, and in particular polyclonal IgG, which comprise, as preservation stabilizer in liquid form, a nonionic surfactant in concentration less than or equal to 0.1 g / 1 and which are essentially free of albumin. By essentially devoid of albumin, it should be understood that no trace of albumin is detected by the Pharmeuropa reference method in electrophoresis on cellulose acetate, which corresponds to an amount of albumin- less than 1% of IgG .
De préférence, la concentration en tensio-actif non ionique est comprise entre 0,02 et 0,05 g/1 environ et est de préférence de l'ordre de 0,025 g/1. De préférence, les préparations selon l'invention comprennent entre 30 et 120 g/1 d'immunoglobulines.Preferably, the concentration of nonionic surfactant is between 0.02 and 0.05 g / l approximately and is preferably of the order of 0.025 g / l. Preferably, the preparations according to the invention comprise between 30 and 120 g / l of immunoglobulins.
Le tensio-actif non-ionique est choisi de préférence dans le groupe consistant en monooleate de sorbitanne polyoxyéthylène (20) , éther octylphénylique du déoaéthylène-glycol, monolaurate de sorbitanne polyoxyéthylène (20) , copolymère mixte polyoxyéthylène/polyoxypropylène et polyoxyéthylène et Laurate de polyéthylene glycol 600.The nonionic surfactant is preferably chosen from the group consisting of polyoxyethylene sorbitan monooleate (20), deoaethylene glycol octylphenyl ether, polyoxyethylene sorbitan monolaurate (20), mixed polyoxyethylene / polyoxypropylene and polyoxyethylene copolymer and polyethylene laurate glycol 600.
Comme cela est d'usage, les préparations selon l'invention peuvent aussi comprendre un stabilisant de lyophilisation usuel tel que le saccharose, notamment en concentration de l'ordre de 50 à 100 g/1.As is customary, the preparations according to the invention can also comprise a usual lyophilization stabilizer such as sucrose, in particular in a concentration of the order of 50 to 100 g / l.
Les préparations d'immunoglobulines selon 1'invention ont également de préférence les caractéristiques préconisées par les normes en vigueur, tel que pH compris entre 4,0 et 7,4, osmolalité supérieure à 280 mosmol/kg par l'addition de solutés osmotiquement actifs, tels que des sels minéraux (tel que NaCl) ou des sucres (tel que glucose, saccharose, maltose) ou des sucres-alcools (tels que mannitol, sorbitol) ou des acides aminés (tel que glycocolle) .The immunoglobulin preparations according to the invention also preferably have the characteristics recommended by the standards in force, such as pH between 4.0 and 7.4, osmolality greater than 280 mosmol / kg by the addition of osmotically active solutes , such as mineral salts (such as NaCl) or sugars (such as glucose, sucrose, maltose) or sugar alcohols (such as mannitol, sorbitol) or amino acids (such as glycine).
Les préparations obtenues répondent aux critères de sécurité d'emploi particuliers aux solutions d'Ig pour administration intraveineuse, à savoir stérilité bactérienne et fongique, apyrogénicité, taux réduit d'agrégats et de polymères et activité anticomplémentaire réduite.The preparations obtained meet the safety criteria for use specific to Ig solutions for intravenous administration, namely bacterial and fungal sterility, apyrogenicity, reduced rate of aggregates and polymers and reduced anticomplementary activity.
Les immunoglobulines présentes dans la préparation peuvent être obtenues à partir de plasma, de sérum ou de placenta par les méthodes classiques de fractionnement des protéines, complétées le cas échéant par des traitements spécifiques visant à réduire le taux d'agrégats et de polymères et/ou à réduire 1'activité anticomplémentaire de la préparation (par exemple traitement modéré à la pepsine ou à la plasmine, traitement dissociant à pH acide, modification chimique par réduction et/ou alkylation ou précipitation des agrégats et polymères par le PEG) .The immunoglobulins present in the preparation can be obtained from plasma, serum or placenta by conventional methods of protein fractionation, supplemented if necessary by specific treatments aimed at reducing the level of aggregates and polymers and / or to reduce the anticomplementary activity of the preparation (for example moderate treatment with pepsin or with plasmin, dissociative treatment at acid pH, chemical modification by reduction and / or alkylation or precipitation of the aggregates and polymers by PEG).
Les préparations selon 1'invention peuvent être des solutions prêtes à l'emploi, donc conservées sous forme liquide, ou être des solutions obtenues extemporanément par dissolution d'un concentré lyophilisé.The preparations according to the invention can be ready-to-use solutions, therefore stored in liquid form, or can be solutions obtained extemporaneously by dissolving a lyophilized concentrate.
La présente invention a également trait à un procédé de stabilisation des préparations d'immunoglobulines humaines ou animales, polyclonales, dans lequel on ajoute à la préparation un stabilisant de conservation sous forme liquide qui est un tensio-actif non ionique de façon à obtenir une concentration inférieure ou égale à 0,1 g/1 dans la préparation finale. De préférence, le tensio-actif non ionique est ajouté en concentration comprise entre 0,02 et 0,05 g/1 environ et de préférence de l'ordre de 0,025 g/1. Le tensio-actif non ionique est choisi parmi ceux indiqués plus haut. L'agent tensio-actif est avantageusement ajouté juste avant le conditionnement final en flacons, mais il peut également être ajouté à un stade antérieur du procédé d'extraction et de purification des Ig. L'invention va être maintenant décrite plus en détail à l'aide d'essais réalisés avec des préparations d'immunoglobulines selon l'invention.The present invention also relates to a process for stabilizing polyclonal human or animal immunoglobulin preparations, in which a preservation stabilizer in liquid form which is a nonionic surfactant is added to the preparation so as to obtain a concentration less than or equal to 0.1 g / 1 in the final preparation. Preferably, the nonionic surfactant is added in a concentration of between 0.02 and 0.05 g / l approximately and preferably of the order of 0.025 g / l. The nonionic surfactant is chosen from those indicated above. The surfactant is advantageously added just before the final packaging in vials, but it can also be added at an earlier stage in the process of extraction and purification of Ig. The invention will now be described in more detail using tests carried out with immunoglobulin preparations according to the invention.
On a simulé des conditions sévères de manipulation de flacons d'Ig intraveineuse à l'aide d'un- test d'agitation. Des flacons de verre borosilicate de type I de 50 ml sont remplis aseptiquement avec 20 ml d'une solution stérile d'Ig à 50 g/1. Les flacons sont agités à température ambiante de 20°C pendant 1 ou 2 h sur un agitateur de type oscillant/alternatif réglé pour 80 oscillations horizontales par minute.Severe conditions for handling intravenous Ig vials were simulated using a shake test. 50 ml type I borosilicate glass vials are aseptically filled with 20 ml of a sterile Ig solution at 50 g / l. The bottles are shaken at room temperature of 20 ° C for 1 or 2 hours on an oscillating / alternating type agitator set for 80 horizontal oscillations per minute.
L'activité anticomplémentaire (AcA) de la solution est mesurée selon le test décrit dans "Pharmeuropa" (supra) .The anticomplementary activity (AcA) of the solution is measured according to the test described in "Pharmeuropa" (supra).
Essai 1 ; effet protecteur du Tween 80 Composition de la solution d'Ig intraveineuse : Ig = 50 g/1 Saccharose = 100 g/1 NaCl = 1 g/1 pH = 4,3Trial 1; protective effect of Tween 80 Composition of the intravenous Ig solution: Ig = 50 g / 1 Sucrose = 100 g / 1 NaCl = 1 g / 1 pH = 4.3
Concentration variable de Tween 80 : de 0 à 100 mg/1Variable concentration of Tween 80: from 0 to 100 mg / 1
Tableau 1 :Table 1:
Tween 80 (mg/1) 0 10 25 100Tween 80 (mg / 1) 0 10 25 100
* AcA avant agitation 0,34 0,35 0,35 0,36* AcA before shaking 0.34 0.35 0.35 0.36
* AcA après agitation 2h >1,19 1,19 0,75 0,38* AcA after shaking 2h> 1.19 1.19 0.75 0.38
* CH 50/mg Ig* CH 50 / mg Ig
Résultat : Une protection nette s'observe dès 25 mg/1. Elle est quasiment totale avec 100 mg/1 de tensio- actif. Essai 2 : effet protecteur de Triton X 100Result: Clear protection is observed from 25 mg / 1. It is almost complete with 100 mg / 1 of surfactant. Test 2: protective effect of Triton X 100
Idem essai 1, mais avec du Triton X 100 au lieu du Tween 80 comme tensio-actif Tableau 2 :Same as test 1, but with Triton X 100 instead of Tween 80 as surfactant Table 2:
Triton X 100 (mg/1) 0 25 50 100 * AcA avant agitation 0,37 0,36 0,36 0,31Triton X 100 (mg / 1) 0 25 50 100 * AcA before shaking 0.37 0.36 0.36 0.31
* AcA après agitation 2h 0,50 0,34 0,36 0,19* AcA after shaking 2h 0.50 0.34 0.36 0.19
* CH 50/mg Ig* CH 50 / mg Ig
Résultat : Une protection totale est obtenue avec la plus faible concentration essayée, 25 mg/1.Result: Total protection is obtained with the lowest concentration tested, 25 mg / 1.
Essai 3 : effet protecteur du Tween 80Test 3: protective effect of Tween 80
Idem essai 1, mais avec une agitation de 1 h au lieu deSame as trial 1, but with 1 hour agitation instead of
2h.2h.
Tableau 3 :Table 3:
Tween 80 (mg/1) 0 25Tween 80 (mg / 1) 0 25
* AcA avant agitation 0,41 0,33* AcA before shaking 0.41 0.33
* AcA après agitation l h 0,70 0,27* AcA after shaking l h 0.70 0.27
* CH 50/mg Ig* CH 50 / mg Ig
Résultat : Dans cet essai moins sévère, la concentration de 25 mg/1 de Tween suffit à protéger totalement l'Ig.Result: In this less severe trial, the concentration of 25 mg / 1 of Tween is sufficient to fully protect the Ig.
Essai 4 ; effet protecteur de l'albumineTrial 4; protective effect of albumin
Idem essai 1, mais utilisation d'albumine humaine au lieu de Tween 80 comme protecteur. Tableau 4 :Same as test 1, but use of human albumin instead of Tween 80 as a protector. Table 4:
Albumine (mg/1) 0 100 1000 10.000 * AcA avant agitation 0,34 0,36 0,26 0,29 * AcA après agitation 2h 1,24 0,64 0,27 0,28Albumin (mg / 1) 0 100 1000 10,000 * AcA before shaking 0.34 0.36 0.26 0.29 * AcA after shaking 2 h 1.24 0.64 0.27 0.28
* CH 50/mg Ig* CH 50 / mg Ig
Résultat : L'albumine exerce un effet protecteur sur l'Ig à partir d'une concentration comprise entre 100 et 1000 mg/1.Result: Albumin exerts a protective effect on Ig from a concentration between 100 and 1000 mg / 1.
Tensio-actifs non ioniques préférés :Preferred nonionic surfactants:
- Tween 80 (ester oléique du sorbitanne polyoxyéthylène fabriqué par Atlas) .- Tween 80 (oleic ester of polyoxyethylene sorbitan manufactured by Atlas).
- Triton X 100 (éther octyl-phénylique du polyoxyéthylène fabriqué par Rohm et Haas) .- Triton X 100 (polyoxyethylene octylphenyl ether manufactured by Rohm and Haas).
- Tween 20 (ester laurique du sorbitanne polyoxyéthylène)- Tween 20 (lauric ester of polyoxyethylene sorbitan)
- Pluronic F 68 (copolymère de polyoxyéthylène et de polyoxypropylène fabriqué par Ugine Kuhlmann) .- Pluronic F 68 (polyoxyethylene and polyoxypropylene copolymer manufactured by Ugine Kuhlmann).
Laurate de polyéthylene glycol 600 (fabriqué par Gattefossé) .Polyethylene glycol 600 laurate (manufactured by Gattefossé).
Le Tween 80 est préféré du fait de son absence de toxicité et de son emploi en formulation pharmaceutique ou alimentaire bien documenté (Voir l'ouvrage "Non Ionic Surfactants" M. Schick éd. Marcel Dekker NY, 1967, 28, 923-970) . Tween 80 is preferred because of its absence of toxicity and its use in pharmaceutical or food formulation well documented (See the work "Non Ionic Surfactants" M. Schick ed. Marcel Dekker NY, 1967, 28, 923-970) .

Claims

REVENDICATIONS
1. Préparations d1 immunoglobulines humaines ou animales, en particulier polyclonales, qui comprennent, à titre de stabilisant de conservation sous forme liquide, un1. Preparations of 1 human or animal immunoglobulins, in particular polyclonal, which comprise, as a storage stabilizer in liquid form, a
5 tensio-actif non ionique en concentration inférieure ou égale à 0,1 g/1 et gui sont essentiellement dépourvues d'albumine. 5 nonionic surfactant in a concentration less than or equal to 0.1 g / 1 and which are essentially devoid of albumin.
2. Préparations selon la . revendication 1, caractérisées en ce gue la concentration en tensio-actif2. Preparations according to the. claim 1, characterized in that the concentration of surfactant
-"-O non-ionigue est comprise entre 0,02 et 0,05 g/1 environ, de préférence 0,025 g/1 environ.- "- O non-ionic is between 0.02 and 0.05 g / 1 approximately, preferably 0.025 g / 1 approximately.
3. Préparations selon la revendication 1 ou 2, caractérisées en ce qu'elles comprennent entre 30 et 120 g/1 d' immunoglobulines.3. Preparations according to claim 1 or 2, characterized in that they comprise between 30 and 120 g / 1 of immunoglobulins.
15 4. Préparations selon l'une quelconque des revendications 1 à 3, caractérisée en ce que le tensio-actif non ionique est choisi dans le groupe consistant en monooleate de sorbitanne polyoxyéthylène (20), éther octylphénylique du décaéthylène-glycol , monolaurate de 0 sorbitanne polyoxyéthylène (20), copolymère mixte polyoxyéthylène/polyoxypropylène et polyoxyéthylène et Laurate de polyéthylene glycol 600.4. Preparations according to any one of claims 1 to 3, characterized in that the nonionic surfactant is chosen from the group consisting of polyoxyethylene (20) sorbitan monooleate, decaethylene glycol octylphenyl ether, 0 monolaurate polyoxyethylene sorbitan (20), mixed polyoxyethylene / polyoxypropylene and polyoxyethylene copolymer and Polyethylene glycol 600 laurate.
5. Préparations selon l'une guelconque des revendications 1 à 4, caractérisées en ce qu'elles 5 comprennent en outre un stabilisant de lyophilisation.5. Preparations according to any one of claims 1 to 4, characterized in that they further comprise a lyophilization stabilizer.
6. Procédé de stabilisation des préparations d' immunoglobulines polyclonales, dans lequel on ajoute à la préparation un stabilisant de conservation sous forme liquide qui est un tensio-actif non ionique de façon à 0 obtenir une concentration inférieure ou égale à 0,1 g/1 dans la préparation finale. 6. Method for stabilizing polyclonal immunoglobulin preparations, in which a preservation stabilizer in liquid form which is a nonionic surfactant is added to the preparation so as to obtain a concentration less than or equal to 0.1 g / 1 in the final preparation.
PCT/FR1994/000955 1993-07-30 1994-07-28 Stabilised immunoglobulin preparations and method for preparing same WO1995003826A1 (en)

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AU73866/94A AU7386694A (en) 1993-07-30 1994-07-28 Stabilised immunoglobulin preparations and method for preparing same

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FR2708467B1 (en) 1995-10-20
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AU7386694A (en) 1995-02-28
CA2167712A1 (en) 1995-02-09
FR2708467A1 (en) 1995-02-10

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