MXPA00010295A - Stabilised virus preparation - Google Patents

Abstract

Stabilised dried pharmaceutical compositions dispersible in aqueous liquid for injection comprise (i) virus e.g. for use as a vaccine or vector, preferably a herpesvirus, e.g. attenuated or genetically disabled infectious herpes simplex virus or varicella zoster virus, (ii) polysaccharide, e.g. dextran, and/or a source of mixed aminoacids of vegetable or bacterial origin, (iii) a buffer, and (iv) a mono- or oligo-saccharide or derivative thereof.

Description

STABILIZED VIRAL PREPARATION FIELD OF THE INVENTION This invention relates to virus preparation, for example, for vaccine or other pharmaceutical or research use, with its stabilization and with processes for producing such preparations, as well as with its use, for example, as vaccines or as viral vectors.

BACKGROUND OF THE INVENTION AND PREVIOUS TECHNIQUE It is known to freeze and / or lyophilize viable virus preparations for laboratory or vaccine use in order to preserve their activity. Many methods are known for producing live viral preparations, for example herpesvirus preparations, for vaccine and other purposes. US 5,024,836 (91.06.18) (Merck &Co. Ine: WJ McAleer et al) discloses a stable, freeze-dried live vaccine of herpesvirus comprising from about 0.5% to about 8% moisture, and claims a virus vaccine. Live attenuated varicella lyophilized injected by gas which comprises 2% to 8% moisture.

Ref: 124193 Documents US 5075110 (91.12.24) and EP 0353108 (Institut Merieux: AJ Francon et al) describe the stabilization of lyophilized attenuated vaccines with lyophilization excipients of amide or thioamide, for example urea. EP 0048194 (Merck &Co Ine: MR Hilleman et al) describes lyophilization processes in which the lyophilization time and expenses are reduced by freezing a liquid vaccine before freeze-drying by rotating the bottle over its side in a liquid bath maintained at a temperature below the eutectic point of the vaccine. The documents US 4,338,335 (82.07.06) and EP 0028563 (Merck &Co. Ine: WJ McAleer et al) describe a stabilizer for liquid vaccines, and a stabilized live liquid viral vaccine containing live virus, partially hydrolyzed gelatin, a monosaccharide or disaccharide, a cell culture medium, L-acid glutamic, L-arginine and buffer to maintain the pH from about 6.0 to about 6.5. EP 0008255 (Merck &Co. Ine: WJ) McAleer et al) describes a herpesvirus vaccine and its preparation, especially a vaccine for the disease of Marek. The virus is lyophilized in the presence of a buffered stabilizer controlled for pH, so that the vaccine can be reconstituted with distilled water. EP 0295043 (88.12.14) (Kitasato Institute: S Makino et al) describes a stabilized live attenuated vaccine comprising at least one live attenuated flat virus which is selected from measles, mumps, or rubella virus and, as an agent stabilizers, lactose, sucrose, D-sorbitol, sodium glutamate and gelatin hydrolyzate. The document EP 0252059 (Smithkline Biologicals S.A .: E D'Hondt) describes stabilizers for attenuated vaccines, which contain lactose, sorbitol, dextran, casein hydrolyzate, L-glutamate, EDTA and buffer at a pH of 6.7-7.2. WO 96/29096 (Hisamitsu Pharmaceuticals Co. , Inc .: H uma et al) describes the production of gene transfer preparations by freeze-drying a mixture of a recombinant viral vector with at least one additive selected from arginine, glutamic acid (or sodium salt thereof) ), serine, glucose, inositol, lactose, mannitol, sorbitol, trehalose and xylose. US 4,985,244 (91.01.15) (Kitasato Institute: S Makino et al) describes a stabilized live attenuated vaccine with improved thermal stability, which comprises a measles, mumps, or rubella virus vaccine, live attenuated plan that grows in a means-199 for cell culture, or a live attenuated vaccine stabilized with lactose, sucrose, D-sorbitol, sodium glutamate and gelatin hydrolyzate. US 4,622,222 (86.11.11) (Phylaxia) Oltoanyagtermelo Vallalat: E Horvth et al) describes a lyophilized vaccine against duck hepatitis using attenuated virus, and its production using infected duck embryos, which include lyophilisate of the sterile viral material with colidone, gelatin, glucose and sucrose. US 4,500,512 (85.02.19) (Institut Pasteur: M Barrae) describes stabilized vaccines containing live virus, especially yellow fever virus, stabilizers comprising phosphate buffer, calcium and magnesium ions, lactose, sorbitol and amino acids that are they select from histidine, alanine, valine, threonine, arginine, methionine, hydroxyproline, lysine, isoleucine, phenylalanine, serine, preferably histidine and alanine. The stabilized vaccine is lyophilized. US 3,985,615 (Osaka Res Foundation: T Kubo et al) describes the production of a live attenuated varicella virus for use as a culture vaccine comprising passage in cobay primary embryonic tissue cells. US 5,024,836 (Merck: WJ McAleer et al) relates to the production of lyophilized vaccine preparations based thereon. US 5,792,643 (97.03.28) and WO 95/10601 (Viagene: SM Hermann et al) describe the preservation of infectious recombinant virus using a saccharide, a high molecular weight structural additive, a buffer and water, and cooling the mixture below the eutectic or glass transition point, and remove water by sublimation to a water content of less than 10%. WO 93/18790 (LK Csatary) describes lyophilized viral vaccines (for example MDV vaccine) with modified starch such as hydroxyethylstarch with a molecular weight of 100,000-300,000 added as a protective colloid prior to lyophilization. The document JP 06234659 (94.08.23) (ZH Handai Biseibutsubyo Kenkyukai) describes a live stabilized vaccine containing live attenuated or recombinant varicella virus and a free stabilizer Ca2 + ion and Ma2 + ion, preferably with gelatin or gelatin hydrolyzate, or a chelating agent such as EDTA. EP 0290197 (88.11.09) (Merck &Co Ine: RZ Maigetter wet al) describes a freeze-dried live herpesvirus vaccine, injected by gas, stable, comprising 0.5-8% moisture allowing storage under standard conditions refrigeration, that is, 5 ° C, instead of lyophilization conditions (-20 ° C). The injection of gas during the primary cycle of the lyophilization process and the drying of higher moisture levels reduces the lyophilization time, typically to 7-11 h for primary and secondary combined cycles. Document DD-209738 (Cent Cerc Bioprep: IV Patrascu) illustrates the production of a herpesvirus vaccine against Marek's disease by: (a) culturing embryo cells on dextran microspheres; (b) inoculate the culture up to 80% confluence with turkey herpesvirus strain FC-126; (c) collecting the infected cells in SPGA medium (sucrose, phosphate, glutamate, fraction V of bovine albumin) when the cytopathic effect is 80%; (d) ultrasonic pulsing and centrifugation to recover a first vaccine crop; (e) resuspending the pellet in SPGS medium and repeating step (d) to obtain a second vaccine crop (to increase vaccine yield); (f) freeze the combined vaccines at -100 ° C before determining the virus titer; and (g) dilute with SPGA medium and lyophilize. The document JP 06234659-A (ZH Handai Biseibutsubyo Kenkyukai) describes, in one example, the production of herpesviral vaccine on human diploid fibroblast MRC-5 cells cultured in MEM medium at 37 ° C; which comprises the inoculation of varicella virus Oka strain of virus seeded at an MOI of 0.03 to MRC-5 cells and culture at 37 ° C for 2 days. The virus is then suspended in a solution containing NaCl, KCl, Na2HP04, KH2P04, sucrose, L-glutamate, gelatin, gelatin hydrolyzate and EDA-3Na. EP 0 573 107, US 5,360,736 and US 5,607,852 (Merck: PA Friedman et al) describe processes for the production of attenuated varicella-soster virus vaccine. WO 98/28000 (Merck &Co., Inc., Rahway, NJ, US: DB Volkin et al) describes vaccine formulations (eg, measles, mumps, rubella, VZV or herpes s.implex) comprising a 6-carbon polycarboxylic acid, disaccharide and buffer. US 3,915,794 (Recherche et Industrie Therapeutique, Belgium: Z Nathan and J Petermans) discloses stable preparations of viruses comprising herpes virus group B (eg turkey herpes virus) and a buffered aqueous solution of pH 6.5-7.5 comprising polyvinylpyrrolidone, sugar , glutamate and chelating agent. US 4,147,772 (79.04.03) (Merck &Co. Inc. - WJ McAleer et al) describes a lyophilized vaccine with a pH between about 6.0 and 6.5 and comprising live virus, partially hydrolyzed gelatin (molecular weight of about 3,000), a 6-carbon polyhydric alcohol, a cell culture medium and acid buffer. US 5,665,362 and WO 92/05263 (Cantab Pharmaceuticals Research Ltd: SC Inglis et al) and US 5,837,261 and WO 94/21807 (Cantab Pharmaceuticals Research: Inglis et al) and the documents mentioned here illustrate previous knowledge related to infectious herpesvirus genetically inactivated such as herpes simplex virus, for example for vaccine purposes and to provide recombinant cells and culture methods for their production. Other descriptions of genetically inactivated herpesvirus and cells to produce it are also included in the prior art, for example certain references indicated in the following. It still remains desirable to provide additional forms of stabilized virus preparations, for example for use as a vaccine.

THE PRESENT INVENTION According to the present invention, there is provided a stabilized dry pharmaceutical composition comprising a virus, which is dispersible in an aqueous liquid for injection and which comprises: virus, for example, as an active vaccine component, for example a herpes virus; vegetable peptone; shock absorber; and a saccharide or a sugar alcohol, or another monosaccharide or oligosaccharide or derivative thereof, for example lactose or sorbitol. The composition may optionally contain dextran or another polysaccharide with a molecular weight greater than 5,000, which, if desired, may replace the plant peptone. Additional optional ingredients may include additional amino acids, for example diacidic amino acids such as sodium L-glutamate or L-aspartate, or a mixture of amino acids. Among the additional ingredients that may be suitable are those referred to in the prior art documents mentioned above, most preferably those of vegetable or mineral origin. In certain examples, the stabilized dry pharmaceutical compositions may comprise: (i) infectious virus as an active component, for example, for use as a vaccine or as a viral vector, for example, for gene therapy, preferably a herpesvirus, for example a infectious herpesvirus genetically inactivated or attenuated such as HSV or varicella zoster virus, (ii) a polysaccharide with molecular weight greater than about 5,000, preferably about 11,000 to about 40,000, and less than 70,000, eg dextran and / or a source mixed amino acids of vegetable or bacterial origin, for example vegetable peptone, for example peptone made by enzymatic hydrolysis of soybean protein, (iii) buffer, for example tris-HCl, bicarbonate, phosphate and / or citrate, and (iv) saccharide or sugar alcohol, for example lactose, sucrose or sorbitol. Certain examples may contain one but not both polysaccharide components mentioned above and the source of mixed amino acids. In addition to the components mentioned above, the composition may contain additional ingredients which include additional amino acids, for example diacidic amino acids such as L-glutamate or sodium L-aspartate, or a mixture of amino acids. Examples of additional ingredients that may be suitable components of compositions of the invention are those referred to in the prior art documents mentioned above, most preferably ingredients of vegetable or mineral origin. Compositions of the invention include free examples of protein (in addition to any protein that is part of the active vaccine component), in particular free of gelatin or other animal protein or its hydrolyzate, or other material of animal origin. When the compositions include a source of mixed amino acids, such as plant peptone, they may be free of materials with molecular weight greater than about 2,000, for example, free of materials with a molecular weight greater than about 1500 (in addition to any material forming part of the active vaccine component). It has been found that lyophilized compositions as described herein can have good titre retention at the end of useful storage periods at moderate temperatures, for example above 0 ° C, for example at about 8 ° C, after the lyophilization of the composition. Under certain test conditions, without limitation, dry compositions are retained for up to 16 weeks or up to 52 weeks, at 8 ° C with an infectious virus titer within 0.5 of a logarithm of the titer found immediately after lyophilization, for example in titers in the range of 10 s to 106 pfu / ml in relation to the volume of liquid before lyophilization. A further aspect of the invention relates to the use of vegetable peptone or other mixed amino acids of vegetable or bacterial origin, free of animal protein or animal protein hydrolyzate, or other material of animal origin, in compositions for stabilizing viruses, and in the preparation of dry stabilized virus compositions, for vaccine and other uses as mentioned herein. The suitable and currently preferred vegetable peptone for preparing compositions according to the invention can consist, for example, of essentially a preparation made from edible solvent extracted soybean meal, and cleaned by hydrolytic digestion with protease, to provide a product with an average molecular weight in the range of about 300-400 and substantially free of constituents of greater molecular weight above about 2,000 molecular weight. Soluble carbohydrate of plant origin may also be present in such a peptone preparation. Alternatively, mixed amino acids of plant or bacterial origin can be used in place of peptone, as described above. The compositions according to the invention can generally be made in accordance with the pharmaceutical practice known per se so that they can reach acceptable standards, for example sterility. The total content of the components in the dry preparation can be such that, when reconstituted with sterile injection liquid, for example water for injection or saline for injection, the composition can be used to provide an injection which is an acceptable approximation to isotonic concentration. An accurately isotonic concentration provides about 330 mOsm, and in accordance with existing practice, it may be an acceptable approximation to obtain this, for example within the range of about 100-600 mOsm, generally within about 250-450 mOsm. "Isotonic" in the present usually refers to such an approach. The dose of virus in a lyophilized preparation according to an example of the invention can be chosen so as to provide, in the liquid for injection constituted, a dose of, for example, about 103 to about 108 pfu of virus. A commonly chosen example of a volume of a dose of for injection is approximately 0.5 ml. The lyophilized preparation can be prepared from a liquid composition which is either at the same concentration in its main components as the liquid to be reconstituted, or in a higher or lower concentration. The moisture content of the lyophilized product may vary from 0.5-15% and may be less than about 10%, for example less than about 5%, for example less than about 2% or less. Also provided with the invention is a process for producing a stabilized dry pharmaceutical preparation of a herpesvirus vaccine, which is dispersible in an aqueous liquid for injection, and which comprises lyophilizing a sterile aqueous composition containing: (i) virus as a active vaccine component, preferably a herpes virus, for example a genetically inactivated or attenuated herpes simplex virus, or varicella zoster virus, (ii) plant peptone as mentioned above, (iii) buffer, for example tris-HCl, phosphate and / or citrate, and (iv) saccharide or sugar alcohol, for example lactose or sorbitol. Optionally, the composition may contain (v) dextran or another polysaccharide, for example, with a molecular weight greater than about 5,000, which, if desired, may be substituted for the plant peptone. The lyophilization of the product can be carried out over any suitable period, in accordance with conventional lyophilization practice, for example at a temperature lower than the vitreous transition temperature of the frozen liquid to be lyophilized, and the product can be form of a solid dry cake within a glass jar, preferably under sterile conditions. The lyophilization process may comprise process steps known per se for obtaining a two stage drying in which a first stage of sublimation of the water content is carried out at a temperature of, for example, about -40 ° C or lower , and then the temperature of the composition is increased to a higher temperature, for example, from 0 to +10 ° C, when the drying to advanced enough so that the cake formed by the partially dry composition retains its shape at a higher temperature , and an additional amount of water is removed during and after such an increase in temperature, even at reduced pressure. In practice it has been found that a reduction in water content below the range of about 2 to about 9% by weight is conveniently and satisfactorily attainable for product stability. The product can be rehydrated conveniently with a sterile aqueous liquid, for example water for injection. Also provided according to the invention is a process for producing a liquid preparation of a viral vaccine for injection, which comprises dispersing or dissolving a sterile lyophilized preparation as specified in the foregoing, for example a stabilized dry pharmaceutical preparation of a virus of recombinant herpes simplex, in aqueous liquid for injection, so as to produce a liquid composition of approximately isotonic concentration. Examples of the present invention are dry stabilized preparations of active herpesvirus, dried from liquid aqueous preparations containing stabilizing agents as follows (w / v): disaccharide 2-12%, for example sucrose, lactose and / or trehalose, preferably at least 2 disaccharides, each in at least 2%; optionally monosaccharide or monosaccharide of sugar alcohol, for example sorbitol at 1.5-4%, optionally dextran at 1-5%; optionally glutamate or sodium aspartate at 0.05-0.7%, and vegetable peptone at 1-4%. The compositions may also comprise other materials such as other colloids, which when present are preferably polysaccharides or polysaccharide derivatives such as hydroxyethyl starch. The virus formulations can generally comprise a live virus, preferably attenuated or genetically inactivated. The virus preferably is an infectious virus, for example a herpes virus, and can be a genetically inactivated virus, for example, by one of the classes described or referred to in WO 92/05263 (Immunology Ltd: Inglis et al. ); LH Nguyen, D Knipe et al., J Virol 66 (12) (December 1992) 7067-7072; WO 94/01573 (Akzo; Peeters et al); WO 94/03595 (Akzo; Visser et al); WO 94/21807 (Cantab Pharmaceuticals Research Ltd: Inglis et al); WO 95/18852 (Harvard College and Dana-Farber Cancer Institute, -D Knipe, et al); WO 96/04395 (Lynxvale Ltd; P Speck); and WO 96/26267 (Cantab Pharmaceuticals Research Ltd: MEG Boursnell et al). The invention is particularly applicable, for example, to herpesviruses and poxviruses, among others. Particularly useful applications are for the stabilization of HSV, for example HSV-2, for example in the form of inactivated HSV-2, as described in WO 94/21807 (Cantab Pharmaceuticals: Inglis et al) and WO 96/26267 ( Cantab Pharmaceuticals Research Ltd: MEG Boursnell et al), for example in modalities in which the virus transports exogenous genetic material that codes for an immunomodulator or a heterologous antigen. Other herpesviruses such as, for example, VZV, BHV and PRV can also be formulated as described herein. Examples of compositions of the invention may comprise, for example, immunogens and vaccines as well as viral vector preparations for in vivo and ex vivo use. The compositions may comprise immunogens different from the viruses described above, for example immunomodulators such as interleukins, for example IL-12; and stabilizers known per se and excipients such as those that may be desired for purposes of a hand-delivered application. A composition such as that described herein can be passed through a sterilizing filter prior to the drying step and can become sterile (in addition to possessing any desired and proposed biological activity such as that of the virus itself). Examples of the compositions provided herein may be made free of constituent materials of bovine origin, and in some cases of another origin or of any animal origin. The compositions provided herein may have useful stability, for example, with respect to the proportion of infectious virus which survives the lyophilization process and / or with respect to the storage stability over extended periods of the lyophilization product, for example during storage at temperatures in the range of about 4-10 ° C, for example about 8 ° C. The invention is illustrated by the following examples which are provided without intending to limit the scope of the invention.

Example 1 A liquid preparation of herpes simplex virus type 2 genetically disabled HSV-2 (for which see specification WO 94/21807, but the invention is also applicable to other viruses), can be lyophilized according to an example of the present invention when dispersing the virus in an aqueous liquid of the following composition (w / v in aq): lactose 5%, sucrose 5%, sorbitol 1.8%, sodium glutamate 0.1%, vegetable peptone 2%, buffer pH 5.5-pH 8, preferably at about pH 7 (about 50 to 100 mM tris-HCl, about 10 mM sodium citrate with additional sodium chloride, for example up to about 138 mM or 10 mM sodium potassium phosphate), and lyophilize the product in a manner known per se in standard glass jars. - Í9 - In variants of this example, the lactose can be substituted by sucrose or trehalose, or it can be omitted. At the end of the storage period of approximately 16 weeks at 8 ° C, certain samples prepared according to this example and in which the phosphate buffer is present, have a titer in pfu / ml (of the original liquid volume before the lyophilization) within 0.5 of a logarithmic unit of the titer found immediately after lyophilization and have a titer of the order of 10 5 to 10 6 pfu / ml relative to the liquid volume before lyophilization. In a variant of this example, tris buffer can be used in place of phosphate buffer.

Example 2 A lyophilized preparation of HSV-2 genetically disabled from the previous example as in the previous example, except that the liquid before lyophilization, also comprises the virus: dextran: 2. 5% (molecular weight of about 11,000 to 40,000 or greater, preferably about 11,000), sodium glutamate 0.5%, sucrose 2.5% and buffer pH 5.5-pH 8, preferably about pH 7, as described for example 1.

In one version of this example, the buffer comprises tissue culture medium M-199 and at the end of the storage period of approximately 52 weeks at 8 ° C, some samples made according to this example have a titer, in pfu / ml (from the volume of the original liquid before lyophilization) within 0.5 logarithmic units of titer immediately after lyophilization, and had a titer in the order of 105 to 106 pfu / ml relative to the volume of the liquid before lyophilization. In the variants of this example, sucrose can be replaced by trehalose. At the end of the storage period of approximately 52 weeks at 8 ° C, certain samples prepared according to this example and in which trehalose is present, have a titre, in pfu / ml (of the original liquid volume before lyophilization) within 0.5 logarithmic units of the title found immediately after lyophilization.

Example 3 A lyophilized preparation of HSV-2 genetically disabled as in example 1 can be made, as in example 1, using components as in example 2, except that the dextran component is dextran with a molecular weight of about 40,000. At the end of the storage period of approximately 54 weeks at 8 ° C, certain samples made according to this example have a titer, in pfu / ml (of the original liquid before lyophilization) within 0.5 logarithmic units of the title found immediately after of lyophilization.

Example 4 A lyophilized preparation can be made HSV-2 genetically disabled from example 1, as in example 1, except that the liquid before lyophilization, also comprises the virus: dextran: 2.5% (molecular weight of approximately 11,000), sodium glutamate 0.5%, sucrose 2.5% and buffer tris 0.1M, preferably pH 7.5. In variants of this example, sodium glutamate can be substituted. This example is protein free, in addition to any other protein that is part of the active vaccine component. At the end of the storage period of approximately 25 weeks at 8 ° C, certain samples performed in accordance with this example present a titer, in pfu / ml (of the original liquid volume prior to lyophilization) within 0.5 logarithmic units of the title found immediately after lyophilization, and have a titre in the order of 10s to 106 pfu / ml in relation to the volume of liquid before lyophilization.

Example 5 A lyophilized preparation of genetically-disabled HSV-2 of Example 1 is carried out, as in Example 1, except that the liquid before lyophilization, further comprises the virus: dextran: 2.5% (molecular weight of about 40,000), glutamate sodium 0.5%, sucrose 5% and buffer tris 0.05M, preferably approximately pH 7.5. This example is free of protein, in addition to any other protein that is part of the active vaccine component. At the end of the storage period of approximately 40 weeks at 8 ° C, certain samples made according to this example present a titer, in pfu / ml (from the original volume of liquid before lyophilization) within 0.5 logarithmic units of the title found immediately after lyophilization, and have a titre in the order of 105 to 106 pfu / ml in relation to the volume of liquid before lyophilization.

Example 6 A lyophilized preparation of genetically-inactivated HSV-2 of Example 1 is carried out, as in Example 1, except that the liquid before lyophilization, comprises, in addition to the virus: dextran: 2.5% (molecular weight of approximately 40,000), sodium glutamate 0.5%, sucrose 2.5% and buffer tris 0.1M, preferably about pH 7.5. In variants of this example, sodium glutamate can be omitted. This example is free of protein, in addition to any other protein that is part of the active vaccine component. At the end of the storage period of approximately 40 weeks at 8 ° C, certain samples made according to this example present a titer, in pfu / ml (from the original liquid volume before lyophilization) within 0.5 logarithmic units of the title found immediately after lyophilization.

Example 7 A freeze-dried preparation of genetically-inactivated HSV-2 of Example 1 is carried out, as in Example 1, except that the liquid before lyophilization comprises, in addition to the virus: lactose, 5%, sucrose 2.5%, D sorbitol 1.8%, 0.1% sodium glutamate, 2% vegetable peptone and 0.05M tris buffer, preferably at a pH of about 7.5. In a variant of this example, sodium glutamate can be omitted. This example is free of protein, in addition to any other protein that is part of the active vaccine component. At the end of the storage period of approximately 25 weeks at 8 ° C, certain samples performed in accordance with this example present a titer, in pfu / ml (of the original liquid volume prior to lyophilization) within 0.5 logarithmic units of the title found immediately after lyophilization.

Example 8 A lyophilized preparation of genetically-inactivated HSV-2 of Example 1 is carried out, as in Example 1, except that the liquid before lyophilization comprises, in addition to the virus: lactose 5%, sucrose 5%, D-sorbitol 1.8 %, 2% vegetable peptone and 0.1M tris buffer, preferably at a pH of about 7.5. This example is free of protein, in addition to any other protein that is part of the active vaccine component. At the end of the storage period of approximately 25 weeks at 8 ° C, certain samples made according to this example, in which sodium glutamate is omitted and the gelatin hydrolyzate is replaced by vegetable peptone, present a titer, in pfu / ml (of the original volume of liquid before lyophilization) within 0.5 logarithmic units of the title found immediately after lyophilization.

Examples of the compositions according to the invention can be lyophilized in a useful manner by the use of standard methods well known in the art for lyophilizing biological material. For example, the following lyophilization process can be used in relation to the compositions of the invention, for example according to the examples given above, and as described below and without attempting to limit the scope of the invention. A virus composition for lyophilization as described herein, is first frozen at -60 ° C for 2 hours, and then dried under reduced pressure at 100 Mtorr using a drying procedure as follows: the temperature of the composition is increased progressively at -42 ° C during the course of one hour, and this temperature is maintained for an additional 60 hours. The temperature is then increased to + 5 ° C over the course of 5 hours, and maintained at that temperature for approximately 7 additional hours. Finally, the temperature is increased to +10 ° C during the course of 1 hour, and maintained at that temperature for approximately 7 additional hours. An alternative and sometimes preferred drying process is as follows: the composition is first frozen by lowering its temperature to -40 ° C during the course of 1 hour, and maintained at this temperature for an additional 2 hours. The composition can then be subjected to a step designed to allow the enlargement of ice crystals: the temperature is increased to -15 ° C over the course of about 45 minutes and maintained at this temperature for an additional 2 hours. The temperature can then be lowered to -40 ° C over the course of 25 minutes and the composition then dried under reduced pressure of 50 Mtorr using a drying procedure as follows: the temperature is maintained at -40 ° C for 2 hours, then it is increased to -15 ° C during the course of approximately 45 minutes and kept at this temperature for an additional 1 hour. The temperature is then lowered to -35 ° C over the course of 1 hour, and maintained at this temperature for approximately 32 additional hours. The temperature can then be increased to + 5 ° C over the course of about 3 hours, and kept at this temperature for an additional 8 hours. In a variant of this example, after freezing the composition, the temperature can be increased to 0 ° C instead of -15 ° C. The invention described herein is susceptible to modifications and variations that will be apparent to readers usually skilled in the art. In particular, and without limitation, processes and compositions of the prior art for vaccine preparations, for example, as mentioned in the documents referred to above, may be applied within the scope of this invention, and this description extends to modifications and variations of the compositions and other aspects of the present invention, including combinations and subcombinations of the features mentioned or described herein and in the mentioned publications and appended claims, which are incorporated herein by reference in its entirety for all purposes.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention is that which is clear from the present description of the invention.

Claims (17)

CLAIMS Having described the invention as above, the content of the following claims is claimed as property:

1. A dry, stabilized pharmaceutical composition characterized in that it is dispersible in an aqueous liquid for injection, and which comprises: (i) infectious, genetically inactivated herpesvirus, (ii) at least one of (a) a polysaccharide or polysaccharide derivative which it has a molecular weight greater than about 5000, and (b) a source of mixed amino acids of vegetable or bacterial origin, (iii) a buffer, and (iv) a monosaccharide, oligosaccharide or derivative thereof.

2. The composition according to claim 1, characterized in that it additionally contains at least one amino acid or amino acid salt, for example L-glutamate or L-aspartate.

3. The composition according to claim 1 or 2, characterized in that it contains a polysaccharide or polysaccharide derivative having a molecular weight greater than about 5000.

- The composition according to any of the preceding claims, characterized in that it is free of animal protein.

5. The composition according to any of the preceding claims, characterized in that it is free of protein, except protein that is part of the herpesvirus.

6. The composition according to any of the preceding claims, characterized in that it contains a disaccharide, for example, lactose, sucrose or trehalose.

7. The composition according to any of the preceding claims, characterized in that it comprises: (i) genetically inactivated infectious herpesvirus, (ii) dextran, (iii) buffer, (iv) sodium glutamate, and (v) sucrose.

8. The composition according to any of the preceding claims, characterized in that it contains a monosaccharide or monosaccharide derivative, for example sorbitol.

9. The composition according to any of the preceding claims, characterized in that it contains plant peptone.

ID. The composition according to any of the preceding claims, characterized in that it contains dextran with a molecular weight in the range of about 5000 to about 70,000.

11. The composition according to any of the preceding claims, characterized in that the buffer is a tris or phosphate buffer.

12. An injectable liquid pharmaceutical preparation, characterized in that it has been prepared by dispersing a stabilized dry composition, according to any of the preceding claims, in sterile liquid for injection, for example water or saline.

13. A pharmaceutical composition, according to any of the preceding claims, characterized in that it is used as a vaccine.

14. A method for preparing a pharmaceutical composition, characterized in that it comprises lyophilizing an aqueous liquid comprising: (i) genetically inactivated infectious herpesvirus, (ii) a polysaccharide or polysaccharide derivative having a molecular weight greater than about 5000 and / or a source of mixed amino acids of vegetable or bacterial origin, (iii) a buffer, and (iv) a monosaccharide, oligosaccharide or a derivative thereof.

15. The method according to claim 14, characterized in that the aqueous liquid comprises a disaccharide at a concentration from 2 to 12%, w / v.

16. The method according to claim 14, characterized in that the aqueous liquid comprises plant peptone at a concentration in the range from about 1 to about 4% w / v.

17. The method according to claim 14, characterized in that the aqueous liquid comprises dextran in a concentration from 1 to 5%, w / v.