WO2012125658A1 - Aerosolized thermostable granular particulate vaccine preparation - Google Patents

Abstract

Disclosed herein are methods for aerosolized thermostable granular particulate vaccine preparations for the administration of a immunogenic vaccine composition composed of viral particles, bacterial cells, microplasmas, prions or other disease causing agents in humans, animals or aquatic species, antigenic products of disease causing organisms such as virus or bacteria, or nucleic acid sequences. The preparation of the present invention provides a stable vaccine preparation that has a shelf-life. The thermostable vaccine is prepared as a granular particulate which can be packaged and delivered to remote areas where sources of electricity and refrigeration are cost prohibitive.

Description

AEROSOLIZED THERMOSTABLE GRANULAR PARTICULATE VACCINE

PREPARATION

10021 BACKGROUND AND F1ELP OF THE PRESENT I VENTIO ::

[004], The present invention relates io vaccines or antigenic compositions and methods of eparation thereof. More specifically, the invention, relates to a method of preparing a vaccine composit n which is thermostable and suitable for the administration of a vaccine for animals.

[006 j, Vaccines com ris g viral particles or bacteria! ceils or proteiaaeaoas antigens roduces by recombinant DNA technology are widely used to prevent disease in humans and an ma as well as in aquaeulture. Ge»e.ra \ virai particles and ac ria fbr use in vaccines are attenuated or otherwise treated with one or more agents so as to lessen or remove their pathogenicity.

Genetic manipulations m be carried oat to produce virus or bacteria of low or absen pathogenicity.

[001], Vaccines have also been developed for mierop!asma mediated diseases, as well as disease mediated through other infectiou agents, including for xam le meta.ioans and protozoans,

[008], It is well known that biological materials, including biological materials in solution, are susceptible to inaetivatio due to heat, oxidizing reagents, salts, etc. Virus particles, bacteria and other infective agents used in vaccines may be readily inactivated after a short period at ambient temperature. Inaetivation may result in loss of in eetivity, compromised inactivity of love vaccines or at low temperature storage, or loss of inmunogenicity. Mmy virus particles, ibr example human influenza vims, human hepatitis viruses and avian bronchitis virus, may only survive a temperatures around 4 ^;C for a short period of tim .

[009]. Vaccines used to immunise humans or animals against disease, generally require storage at temperature of 4 or less, such as -29^*C. Such vaccines may only be stable ibr a relatively short period of time, such as 2 to 14 days after production. The requirement ibr low temperature

I storage creates handling am! transport problems. Low temperature storage is costly. Short pe iods of vaccine activity even at low temperature limit vaccine use and raise costs ther by limiting accine distribution particularly in under developed third world, countries,

(OO IO). Globally, market fo vaccine market has bees estimated as exceeding 20 billion dollar in 2010, The majority of li ve viral or bacterial vaccines for ose is human beaJih are attenuated, such as being non-pathogetue strains, or strains of limited pathogenicity, fox example produced by rec mbin nt DNA technology or other meaus. Other commercially available vacc na include viral or bacterial proteins, or proisin/peptides, derived therefrom, such as epitope vaccines containing peptide, protein, glycoprotein, or other epitope of disease c using virus, bacteria or othe organisms, optimally associated with a carrier such as further peptide, protein or other ageiit(s), for example by oovalen bonding or other association.

I 1 i l lJ M mMM && 00i2 , Matty commercially available vaccines, osed for example in feuman and animal health, are whole particle or cell-based products in order to provide m ximum protective properties following vaccine administration. Such vaccines may be live vaccines of attenuated or absent pathogenicity. These products have strict refrigeration requirements is mentioned above, and accordingly a short shelf life.

(0013). Freexe-drying under vacuum (iyophilisation) has bees proposed to prepare vaccines. For exam le, EP-A-29 19? describes !ree¾e~dr!ed tetravalent vaccines,

|CK 1 j. Fri^>e:ze-drymg processes iraditiosaMy involve freezing a solution containing as

Immunogen, such as virus particles, bacterial cells, or protemaeeons antigens and converting ice crystals into water vapor under vacuum {sublimation}. Unfortunately, such processes can damage the native structure of proteins, disrupting viral particles of bacterial cells. Thus a.

detriment l, effect on the imnianogen may result, compromising or destroying inimnsogenieity,

[O0J SJ. Free e-drying is also a complex process with a number of variables, and ma be difficult to perform in a reproducible manner. [00 6], Another problem with freeze drying in the field of vaccine production s the processing requirements, it is not possible to concentrate high doses of vaccine material in small volume, indeed, it is important in the

process thai a large surface a ea of fluid is available to be in contact with the acuum. As only the top of a ftossen volume of material is in contact with the vacuum, vaccines are required to be iree¾e-drled in large containers providing maximum surface area for lree¾e«-dr ½g. The necessary apparatus lor such processes is therefore space consuming and inefficient. As a. consequence if eexe-dried vaccines are generally expensi ve t produce,

[0017], U.S. Pat, No, 5,616,329 describes a process where an aerosol of a microbial suspension s exposed to elevated temperatures such that only the heat stable components of the microbial suspension retai their immunogenic properties. The heat Inaetivatlosi step according to U.S. Pat. Ho 5,61 ,329 emp oys, temperatures in the range of 1 0 to 160 degrees Celsius. The iramunogenieity of the suspension components that are unstable at higher temperatures is lost according to this proposal, and thus this process is unacceptable in tmny vaccine applications,

[0019]. To overcome the limitations of the prior ar and address various problems in the field of vaccines, including cost and production difficulties, handling an storage limitations, and vaccine stability and maintenance of imronnogeniclty, the present Invention disclosed herein provides a process to create a thermostable granular _.particulate for the administration of a 11 VT vaccine. The metho o f the present In vention for the production of a vaccine comprising one or more im nunogens, such as viral particles, bacterial cells,, micro-plasmas, prions or other disease causing agents In humans, animals o aquatic s ecies, antigenic products of disease causing organisms such as virus or bacteria, or nucleic acid, sequences. The process of the Invention stabilises imr ogens from inaetivatlon and loss of imniunogenicity.

[fM 20|. in accordance with one embodiment of the present invention there is provided a process for the production of s a il is d vaccine composi tion of .i nimunogens, wherein the method of the present invention comprises granular particulate of oiannitof hydroxy propyl cellulose (HFC) and magnesium stesrate thai is aerosolked with an antigenic viral culture providing thermostable, inexpensiv and easily packaged vaccine preparation. Some preferred mode of delivery for the vaccine preparation of the p esent invention ma be pressurised metered dose inhalers (MD¾ dry powder inhalers (DP!), nebulizers, sublinguals, skin s ra s, and dental sprays,

[00*!], in accordance with anot e aspect of the a ention there is ro ided a thermostable dr powder particulate, preferably stable at am ie t t m er tures, comprisin the above described iramunogeo. coated particles, the composition having between about 0.1% w/ about 10% w/w. Als preferably the mode of delivery rosy include oral granules which can be dissolved directly in the mouth, cold or room temperature foods or liquids, or prescribed in a capsule or tablet form. 0032], Preferably, the srnrauoogen comprises virus particles, bacterial ceils, other

microorganisms, e karyotie cells, or antigenic products thereof. The irnnrunogen ma contain two or more different virus particles, bacteria celts, other microorganisms, or antigenic products thereof etc. so as to give a multivalen vace sie composition,

[0023], Preferably the virus particles include one or more of animal aad human viruses.

(00.24), Examples of avian viruses include: turkey herpesvirus (!-IVT), arek's disease virus (visceral leucosis), chicken influenza virus, Newcastle disease, avian rhino tracheitis vims, avian herpes virus, fowl pox virus, avian encephalomyelitis Infections bronchitis. Infectious Bursal disease (Girmboro), avian reovirus, fowl laryngotraeheitis an Egg Drop Syndrome vims,

(00.25), Examples of porcine viruses include: Porcine reproductive and Respiratory Syndrome, foot and mouth disease virus, porcine influenza virus, porcine parvovirus, psendorabies vims, nd porcine rotavirus and swine influenza virus,

[0026], Examples of feline viruses include: feline herpes virus, feline inmtunodefieiency virus, feline leukemia virus, feline panSettfcoperna, feline eaheivirus, feline viral rhinoiracbeitis, Mine eoronavirus and rabies,

(00.27]. Examples of canine viruses Include: canine distemper virus, canine adenovirus, parainfluenza, and canine parvovirus, canine hepatitis virus, canine herpes virus and rabies. [002 S|..Exam les of equine viruses include: equ ine encephalitis virus (Easter, Western and Venezuelan equine viral encephalomyelitis), equine infl enza, nd equine herpes virus (equine r!unopnenmooitis),

10 2 1. Examples of bovine viruses include: infections bovine rhinotmeheifis, bovine viru diarrhea virus, bovine respiratory syncytial vims, eoronavirus, and foot and mouth disease virus and paramfluerim.

[0030J. Exam les of human viruses include: hepatitis A virus, hepati tis B vi us, hepatitis C virus, e pes sim lex virus type I and type 2,Va:rieelk-X ster virus, cytome alov us, Epstein- Barr virus, human herpes virus 6 and human herpes virus 7, influenza virus, respiratory syneital virus, parainfluenz virus, adenovirus, rhinoviroses, human immunodeficiency virus, ientiviruses, human papillomavirus, measles virus, mumps virus, ol o vims, rubella virus, human rotavirus, pox virus (such as small pox virus, arbovirus transmitted disease such as Japanese Encephalitis, tick-borne encephalitis, rabies virus, yellow fever virus. West Nile virus and dengue virus.

[0031 ], Preferably, vims particles are live. As vaccines com os te according to this invention are tree flowing powders, vaccines containing different immonogens can be simply blended together free of compatibility problem which may otherwise arise with conventional liquid vaccine admix tures, Thus a preferable aspect of this invention is multivalent vaccine compositions containing tw or mom different vaccine compositions.

(^'0O32J. Preferably, the bacterial cells comprise one or more bacteria from bacterial genus, .including; Escherichia _> such Bs-cherichw cdli including enterotMig nlc, ente⁾pafhogenie, enteroisvasiee and euterouggragative E, mil: afm mHa such as Salmmeila Typhi and Salmonella eniwiikiis, Haemophil , such as Haemophilus influenzae includingH, iw0mn Serotype B, MaetmpbiUs petrosals, Ilmmophills somm and Haemo htlis pa gal nar (Infectious my ), Chlamydia , such as Chlamydia pneumonia ami Chlamydia trachomatis, ^' eisseria such as Neisseria meningitidis. Vibrio such, as Vibrio choleme G roup A. and G ou B Streptococcus such as Streptococcus pneumonia (pneumoeoeeus) and Streptococcus .mis, [jughmelh, such as Legionella pneum phila, Bacillus, such as Bacillus ihmxis,

Mycobacterium, such as Mycobacterium leprae and Mycobacterium parafuherculosis_^ Clostridium, such as Clostridium hohtntum, Clostridium femani, C sirid m prcfHitgengem and CImiridmm Difficile, Pastetmlia, such as. P tmreih ultock and Pm elfa

camofyttca, Bonicieii , $«eh as Bordetella brmchisepii mid B kt lfa p&imsis,

Acii kiCiihfS such as Actmobacil s sui< and bacteria including Bty pei&flmx tkusktpatkw$_t Leptospira,

diphtheria.

(0033). Mycoplasma such as Mycop!a mo kypmmmonkfe. Mycoplasma gaUkepiicum,

Mycoplasma synovim and Mycoplasma pneumoniae mm be used, in this invention. 3 1, Preferably the imnnmogen comprises antigenic products frem disease causing viruses, bacteria, and/or othe disease causing microorganisms. Such antigstsie products include viral sub-particles, viral particle* without their nucleic acid content, viral proteins, bacterial proteins, bacterial I i ^polysaccharides, glycoprotein, carbohydrates or two or more of the aforementioned aniigsn.se products, Antigenic products may be epitopes comprising a sequence of amino- acids, or polysaccharides, antigens produced by re om ant DNA technology or the like, derived from viral and/or bacterial proteins and or carbohydrate and/or lipid seq ences, optionally conjugated to a carrier, such as a peptide or protein,

100351- Preferably the vaccine composition is a free flowing particulate composition. 00361 Preferably the inirnunogen costing of the present invention includes additional constituents such as amino acids, proteins, chelating agents, buffers, preservatives, stabilisers antioxidants, eniulsiilers, p!asttci¾crs and lubricants, 0O37J. Preferably the imrnunogeo coating includes an adjuvant such as aiumiruim salts (alum)., niuranwl peptides and analogues or derivative, saponins (for example Quillaja saponin) or saponin containing compounds, polynucleotides or synthetic nucleic acid, derivative such as polyribonucleotides, sulfur-containing compounds suc as Divema and ploronic po!yols, amine and !ipid-contaiuing compounds, avridine, di.n^thy?-^ociade«y1~ammom«y.m bromide, polyphosphate, cytokines (suc as Interferon) or biodegradable water so ol emulsions such as emulsified paraffin. Other adjuvants or agents with, inimunostsmuiatioa. or immi mo uJa ing o antigen presenting properties, and commercial products Impran, Emunade, Emnlsigen or Aniphtgen may also be used. [0038], Preferably, the vaccine composition is a live vaccine, that is, it«munogens are capable of inducing immune res o ses by mimicking natural virai infection in \ immunised u jec - Live vaccine compositions m y be stable for p to 30 days or more at 4^*€ and at ambient temperature, for ex mple, at 2S^*€, For example, where the i mutiogem are virus particles, the virus particles m be live arid Infective at the c mpletion of the process, yet stable at room temperature storage. Live accine compositions so produce may be stable for periods up to 30 clays or more stored at an ambient temperature, f r e am le_* at 25 X,

[0039], Preferably the fluid comprising one or more i «5««oge»s contains a suspension or dispersion of inm nnogens, such as viral particles, bacteria! ceils or other mieroorpnisms, eukaryotlc cells, or antigenic products or viral particles, bacterial cells or other microorganisms.

[0040]. The fluid comprising one or more immunogens may be a culture medium or other aqueous fluid or media containing the immonogen. The fluid may include one or more additional constituents, such an amino acids, proteins, chelating agent, buffers, salts, preservatives, stabilisers, antioxidants, emulslfiers, p!asilcizer or lubricants.

[00 1 J , Detailed Description

(0042). Tins invention in its various embodiments pro ides processes for the production of stabilised vaccine compositions. The processes of the Invention are suitable for the production of vaccines containing viral, particles, bacterial cells, mycoplasmas, antigenic products, prions or other disease causing agents in humans, animals or aquatic species. The process of the invention provides, in one preferred embodiment high potency live vaccines, stable at room temperature for xtended periods, and processes for their roducti n. Such vaccine compositions have itherto been uoknoxvn.

[0043], in accordance with one aspect of this invention there is provided a process for the production of vaccines comprising one or more immunogens, such as viral particles, bacterial cells, mycoplasmas, prions or other disease causing agents m humans, animals or aquatic species, or antigenic products thereof The process of the Invention stabilises immunogens from inactlvation and loss of immunogenic ity . In this aspect the invention provides a process for the production of a stabilized vaccine composition of immunogens, particularly labile immunogens. wherein oral r mmes are aerosol z d with an antigenic viral culture a a temperature of about 25 ^* C to about 50 , preferably at about 30 C„ such that the imrnunogen coats and is dried onto the particles, The vaccine product containing particles having a moisture content between about 0.1% w/w and 10% w/w so as to give a stabilized vaeciue composition.

[0044] . The immnnogen may comprise virus particles, bacteria! cells, other microorganisms or eufcaryoue cells or antigenic products thereof. The imrnunogen may contain two ox more different virus particles, bacterial ceils, other microorganisms or antigenic products thereof so as to give multivalent vaccines. 0045], Any type of virus particle, or bacterial cell, mycoplasmas, prions or other disease causing agents in humans, animals or aquatic species, or antigenic products of vims particles, bacterial ceils, mycoplasmas, prions or other disease causing agents may be used In th¾ invention.

[0046], Preferably, the imrnunogen comprises virus particles. Preferred virus -particles Include: M^'VT arid avian viruses; porcine viruses; feline viruses; canine viruses; equine viruses; and bovine viruses; hepatitis A virus, hepatitis B virus, hepatitis C virus; fcerpes simplex virus type 1 and type 2; Varleelia-Zoster virus; cytomegalovirus; Bpstein-Barr virus; human herpesvirus 0 and human herpesvirus 7; ktluen¾a virus; .respiratory syneital. virus; parainfluenza virus;

adenovirus and rhinoviruses; human immunodeficiency virus and ieniivlrnses; huma

papillomavirus; measles virus; mumps virus; polio virus^'; rubella, virus; human rotavirus; pox virus (such as smallpox virus); arbovirus transmitted disease such as Japanese Encephalitis; tick- home encephalitis and rabies virus; yellow lever virus; West Nile virus; dengue virus.

[0047]. Preferably, the bacterial cells comprise one or more -bacteria horn bacterial, genus, including; Escherichia_* such Escherichia cott including enterotoxigenic, e ter paihogenle, euteroinvasi.ee and enteroaggragaiive £. coU ; SalmmeHa, su h as Salmonella Typhi and

Salmonella enteritidis, Haemophilus, such as Haemophilus influenzae including II influenza- Serotype B, Emmopkilis parasms, Hoemophilis so s and tiaemaphitis p&rvtgaUinantm

(Infectious Cory a), Chlamydia, such as Chlamydia p monia and Chlamydia trachomatis, Neisseria such as Neisseria meningitidis, Vibrio such as Vibrio choieme Group A and Group 8 Streptococcic such as Streptococcus pneumonia (pneurwocoocus) ami Strepiacoccus mis, Legionella_* such as Legionella pneumophila_* Bacillus_* such as Bacillus anthmxte M cobacterium_* such as Mycabactermm leprae sod Mycobac iwn paratiibercuimis_*

Clostridium, such 88 C strid m botutkmm, Clostridium tentam_< Clostridium prek gmge and Clostridium D ktte, Pastewe!k, such as PasteureJla muttocida and P ier e!

hmmoiytim. Bordeieiia, such as Bordeteiia hm hiseptica and Bonkmi pertussis,

AcUttobaeittns such us Acti kwU!m snte_* md bacteria includin Bryaip lathrix rhmiopaik e, Le tos ira, Borrelta burgdorferi, Helicobacter pylor and Coryttebacf um diphtheria,

[0048], Myct>pkxsm& such as Mycoplasma hypone moniae_* Mycoplasma gelliseptieum_* Mycoplasma synoviac and hfyeoplastna pneumoniae may be sed, in tins invention.

[0049]. Virus particles, bacterial cells, other microorganisms or still other immunogens ma fee al ve or intact, thai s not killed by hea treatment or oilier processes. The processes according to a preferred embodime t of this invention are adapted for the production of vaccine compositions containing liv inirnunogens, such as viral particles.. On administration to a subject, whether human, animal or aquatic species, the vaccine compositions containing live virus particles or other immunogens elicit a strong immune reaction.

[0050]. Alternatively, viral particles, bacterial ceils, other microorganisms, or still other iramunogens, may he killed; that is, heat treated or otherwise treated such that they are not capable of reproductioa in a hosi. Subuni.es or other antigenic products, tor example of virus particles or bacterial cell fractions, may also he used in the invention, as may antigens thereof, such as peptides, proteins, carbohydrates, lipids, llpoixdysaceharides, glycoproteins, or two or more of the aforementioned antigenic products. Antigenic products nsay be epitopes comprising a sequen e of amino acids, o polysaccharides, or the like, derived, from viral and or bacterial proteins and/or carbohydrate and/or lipid sequenees_:! and optionally conjugated to a carrier, such as a peptide or protein.

[00511. In an ahernsti ve embodiment the Inimnnogen ma he a nucleic acid se uenc , such as DMA or NA, lor example based on viral or bacterial or other microo nism nucleic acid, sequences, -which may, for example, be delivered i viral vectors such as pig or fowl adenovirus or fowl pox virus or other vi al vectors suitable tor stabilization according I» the present invention. [0052], According to one aspect of the present invention, one or more immunogens are provide in an aerosol The aerosol preferably comprises a sus ension or disp sion of mimuno ens, sudh as viral panicles bacterial ceils or other microorganisms or eukaryotie cells. The aerosol may be .cu!iare m dium or other laid and media containing the in iunogen, optionally diluted with a diluents in which immunogen is stable (that is, not. inactivated}. Diluents are well k own in the art of virology and microbiology, and include, for example,, sterile water, phosphate buffered saline (PBS), tri buffered^* saline (IBS), sterile water containing sucrose and skim niOk, and 90% sterile water containing sucrose and sk m milk (an example being 5% of both sucrose and sk m milk, and 90% sterile water). Diluents may also includ one or more of gelatin, dexten, EDTA, amino acids such as glycine and egg albumin, mineral salts such as magnesium, sulphate, calcium chloride, and calcium phosphate, and the like,

[0053 J, Chemical .moieties suitable for derivapxatlon may be selected from among water soluble polymers. The polymer selected should be water soluble so that the component to which it Is attached does not precipitate in an aqueous environment, such as a physiological environment. Preferably, for therapeutic use of the end-product preparation, the polymer will be

pharmaceutically acceptable. One skilled in the art will he able to select the desired, polymer based on such considerations as whethe the polymer/componen conjugate will he used d-serapcutically and if so, the desired dosage, circulation time, resistance to proteolysis, and other considerations. For the present component or components, these may be ascertained using the assays provided berem.

[0054] . The wate soluble polymer may be selected from fee group consisting of for example, polyethylene glycol copolymers of ethylene glycol/propylene glycol, carhoxymethylcellniose, dextran, polyvinyl alcohol polyvinyl pyrrol tdone,. poly-! , 3-diox !ane, poly _:3_>r- rioxaoe_s ethyleue/rna!eic anhydride copolymer, polyamfooaeids (either homopolymers or random copolymers), and dex rao or po!y(n~vinyi pyrroiidooe) polyethylene glycol, propropylene glycol homopol mers. pro!ypropyiene oxtde eihy!ene oxide co-polymers, po!yosyethylated p lyols and polyvinyl alcohol. Polyethylene glycol propionaSdenhyde may have advantages in m nufac uring due to its stability In water. [0055] . The polymer ma fee of any molecular weight, and may be branched or ooonmehed. For polyethylene glycol e preferred molecular weight is between about 2 kDa and about 100 kDa (the term "abouf indicating that in preparations of polyethylene glycol, some molecules will weigh more, some less, than the slated molecular weight) for ease in hand ng and

manui cturiag. Of fer sixes may be used, depend ng on the desired therapeutic profile (e.g., the duratioa of sustained release desired, the effects, if any on biological activity, the ease in handling, the degree or lack of antigenicity and other known effects of the polyethylene glycol to a therapeutic protein or analo }.

[0056], The number of _.polymer molecules so attached may vary, and one skilled its the art will he able to ascertain the effect on function. One may mono-derivative, or may provide for a dk irk terra- or some combination, of derivntization, with the same or different chemical moieties (e.g.., polymers, such as different, weights of _.polyethylene glycols). The proportion of polymer molecules to component or components molecules will vary, as will their concentrations in the reaction mixture. In general, the optimum rati (in terms of efficiency of reaction in that there is no excess un.reaeted component or components and. polymer) will he determined by factors such as the desired degree of derivatlzatlon (e.g., mono, d ¾^"k etc,}, the molecular weight of the polymer selected, whether the polymer is branched or unbrauehed, and the reaction conditions.

[005?]. The polyethylene glycol molecules (or other chemical moieties) should be attached to the component or components with consideration of effects on. fxmeiiona! or antigenic domains of the protein. ^"There are a number of attachment methods available lo those skilled in the art, e.g.. EP 040 ! 384 herein incorporated by reference (coupling PEG o G-CSP), see also Mahk ei aL, 1992, Ex . Hematol. 20:1028-1035 (reporting pegylation of Cl -CSF using fresy! chloride). For example, polyethylene glyeol may be covaieotly bound through amino acid residues via a reactive group, such as, a fiee amino or carboxy! group. Reactive groups are those to which an activated polyethylene glyeol molecule may he bound. The amino acid residues having a free amm group include lysine residues and the terminal amino acid residues- those having a free earbo.yyl group include aspartse acid residues glutamic acid residues and the C-ferminal amino aci residue, Sn!fhydrl groups may also he used as a reactive group for attaching the

polyethylene glycol molecute(s). Preferred for therapeutic purposes is attachment at an amino group, such as attachment at the -fermkos or lysine group. [0058]. in some emboctifncata, a composition s in a unit dose formuktion for oral administration to a patient, in some embodiments., a unit dose of the corticosteroid is administered from a mete ed dose device. In some embodiment, the metered dose device delivers a metered unit dose of a composition described herein to the outh or throat, of an individual in need thereof. In certain embodiments, the mete ed dose device is a metered inhaler, which is utilised to administer a metered unit dose to the mouth or throat of an indi vidual (the indi vidual swallows rather than inhales the metered uni dose). In some embodiments, a composition or unit dose described herein is administered as a nebulised comp s tion, an aerosolized composition, an emulsion,, a soiutkm, a suspension, a syrup, a siurtv, a dispersion, a colloid, & dissolving tablet, a dissolving wafer, a capsule, a gel capsule, a semi-solid, a solid form a gel, a gel matrix, a cream, a paste, or the like.

(^'00S J. Table 1 shows e am le of vaccines which may be used In the present Invention include those from the following manufacturers;

[00601. Tabl 1

[00611 Albumin (Human) None j Albumin tteaphanna harfnazeimka

(Iferaani Prodakt!onsgesnrbii

Araias; NP ^' Baxter Healthcare Corporation ra!asf Baxter Healthcare Corporation

[0062 j. Alpha! -Proteinase inhibitor

t SLASSIA' Kanvada Ltd.

(Hum n)

feilasiitr Talecris tliotherapenties, !no

Pro stin-C* alecns Bioth rapeatics, inc.

Zen¾ora ' Aventis Behrlng !, t .( ^'

[00631. Anthrax Vaccine Adsorbed Emergent Bl Defease

Bioihrax⁸

OpsraSiorss Lansing, Ine

[00641, Anti-8 (Murine :

None f Anti-B (Murine

Monoclonal) fnimneor, !ne

Monoclonal}]⁹

[00651- Antibody t Hepatitis B Genetic Systems RSsAg Bio-Rad Laboratories

ASph «ate^i!>

Grobi-I

Recombinant)

! 00109). Hepofiti* B Virus

1 (Hepatitis S Virus/1½tyrnerase

COBAS Am HSereen

Chain Reaction/Blood Cell Roche Molecular Systems, !nc j

BBV Tcei*'

Derived)

1 [00110]. Hepatitis € Virus

(Hepatitis C V.ir«sPoi mer¾s«

COBAS AmptiSeteen

Chain Reaction/Blood Cell Roche Molecular Systems, Inc j

HCV Test, version 2.0'*

Derived)

[00! i I]. He atitis C V irus

Enc ed Antigen (HCV Encoded

A«tigenEn.£yme Immune Assay ABBOTT HCV EIA 2,0^ί¾ί Abbott Laboratories

1 (B1A}Rec©ra mant)

[00112], Hepatitis C Virus

Encoded Antigen (HCV Encoded

Anrige Enx me immune! Ass y Oriho HCV Version 3,0

Ortho-Clinical. Diagnostics, ELISA Test Sysiersr^>!

(EIA), Version 3,0Reecnifein¾it Inc.

and Synthetic)

[00113). Hepatitis C Virus

Encoded Antigens (Recombinant

ABBOTT PRISM HCV⁷¹ Abbott Laboratories

el 00-3, MCr43, S5) i [00114). Bepadtis C Virus

Encoded Antigen CHIRON RIBA HCV 3.0 o>f srii Vaccines and i (lecombmantSyntbetic) RiBA) Strip immunob!oi Assa '" Diagnostics, Inc

IS

1 (HCV) RMA and Hepatitis B Virus

1 (IBVi DNA Assay

1 [00121],. Human

i immun deficie cy Virus Tvpe 1

and/of Hepatits C Virus and/or

j l! a¾its B Vims (HIV - 1 and

Hepatits C Virus and f fcpaun.s B Proeteix tiltrio Assay**^' CJeft-Pro e, i.uc

i Vims/Nucleic Acid Fooled

Testing/Synthetic)

j [001221. Boman

l Imman defi ieacy Virus Type 1

ami / or Hepatitis€ ims (HlV-t Proelek HJ V-1/ CV

Oeo-Probe, toe

arid Hepatitis C Virus / Nucleic Assay⁸'

Acid Pooled Testing / Synthetic) j [001.23 j. Human

1 Immimodefieiency Virus, Type .1 None [Human

1 mmn u d rl eieney V i rus.

i (HIV-l) Reverse Transcription Type I (HIV ) Reverse

BicUie Plasma Services, U\ j(RT) Polymerase Chain Reaction Ί rassctiption (RT

Polymerase Chain

j (PCR) assay Rea ti n (PCR) ass y^"] i [00124). Human

imnmnoddkiency Virus Types 1

and 2 (E. coll B, megater turn, ABBOTT PRISM HIV O

Ab»«ParkJL

1 ecombinant) Antigen and Plus^?:i

j Synthetic Peptide (ChLiA)

Flebogairana^ Instititto kit is, SA

Vaccine Fluam^HK? 1 GlsxoSrnMhfCline BiologkaSs

, ,ij< i ¾) Biomedical Corporation of Hui.aval ^' ,, .

i Quebec

Novarfe Vaccines and i¾vinn^':¾>

Diagnostics Limited

Vlxi m and Pteo«e

Simo.fi Pasteur, f nc

High-Dose'⁸'

[00139;. Inilaeuza im

o e j I fluenza Virus _tv i- <^■> _$

Vaccine, Π5Ν1 , r ■ _{? K}-v , a* , I Sa«o.h Pasteur, lac

Vaccine, fbh 1

1001 0) Influenza Vims

Vaccina l e, Intranasal FksMisr^!W Medlmrenae Vaccines-, ine

[001411 Ja anese

Research Foundation for Encephalitis Virus Vaccine

.IL-Vax^niI Microbial Diseases of Osaka

University

10 1 21. Japanese

Encephalitis Vaccine, i salivated.

iXI.ARG^m 1 mereell 81 oroedics 1

Adsorbed

[00I43J, Measles, Mumps

and Rubella Virus Vaccine, Live M-M- H^Sii Merck & Co, trie

1 [00144]. M easles Mumps,

Rubella and Varicella Virus

PoCHmd'" Merck & Co, inc

Vaccine Live i (0 145 J. Measles Virus

; Vaccine Live _Λ ^>ΠΈ θνΑΧ 1 Merck & Co, fee (0 1 6). Meamg cocct! eaveo"* Novarfe Vaccmes and

Diagnostics, inc.

(Groups A, C_$ Y, ami W-BS)

Oligosaccharide Dipfi&efia

CRM 197 Conjugate Vacdae

[001 7] Meningococcal

Groups (A, C, Y\ md W-135)

Polysaccha ide Diphtheria Toxoid Meaactra'¹ Sanoii Pasteur, inc.

Conjugate Vaccine

[00148], Menin ococcal

Polysaccharide Vaccine, G roups A, Mcncmune-A/C/Y/W-

Sasiofi Pastcu.', inc

C, W-BS Combined I 5

[00149], OLYMPUS

None OLYM US

P System Biood Grouping and PK System Blood

DiAGAST

Piienctypmg Reagervis Grouping and

P enotyping Reagents⁵ '*)

[001501. Pnearnoeoceaf

Vaccine, Po!yvitlcoi FriC«mo x23^SJ-⁵ Merck & Co, inc f OISl). Pneumococcal 7- valeut Co ju ate Vaccine

Prevnar^;-^>; W eth Pharmaceutie ls, inc (Diphlheria CRM 1 ? Protein)

(00152). Pneumococcal. 33- vaient Co jugate Vaccine

Prevnsr 13"^; Wveth Pharmaceirticsls, i nc (Diphtheria CRM 197 Protein)

(00153). Pohovims Vaccine i OL^m Sanofi Pasteur, SA

100.173 Vaedms Immune

Globulin loirave.«w« (Human)

Γ0Ο174Ί Varicella irus

Vaccine Live Varivax¹⁴' \ Merck & Co, Ine

1001751. von WUiebrand

1 Factor/Coagulation Factor VII I Ociaih mia

Wiiaie^¾ %

Complex (Hurnai Froduki osges, m ,h, B . [00?76j.. West Nik Virus co as TsfSeroeo West

Nile Virus Test for use \_a . ... , < „ , (WNV) R A wuh »¾* Ci sbus s lin

s stem⁵ ^

j |00177], West Nile Virus

\ (WN Nuefek Acid Pooled

Prodek West Mile Virus

i Tesiisig/Sy«fheic) Geti-Probe, Inc

(WNV) Assay ^m

||;001?8|. Yellow Fever

\ Vaccine YP- ⁵ Sasiofs Pasteur, lac

{ 00179}. Zoster Vaccine

stavax^!;> Merck & Co, iac

[00180 Animal Vaccines

[00 i 8 i J . Anaplasmosis Vaccine

[00182], Anthrax Spore Vace \

[001 S3] . Arthrohaeter Vaccine

[0 184). Autogenous Vaccine [001 SSJ . Avian Encephalomyelitis Vaccine

1(00186). Avian Eacephalemyelitis-Giickea Anemi Vkas-Fo i o Vaccine

[00 ! 87], Avian Eiicepha!omyeiitis- l Fox Vacci

[00 ί 88] . A ian E»cepImIoRiyeMiis-Fowl Poxiar ngotraeheilis Vaccine

[00189] , B iueloagwe Vacc ine

(001 0) . Bordetdla Avium Vac ine

[0 1 1 ]. BonleteiSa Brone isepiiea Vaccine

[00192) . B« ½e Farai flwesm 3-Ritspiratory Syncytial Virus Vacc ne

(00193 ) , Bovine Respiratory Syncytia! Virus Vaccine [00194]. Bovine R teoirache¾is Vaccine

(00195). Bovine Rhi»ot3¾c eitis-~Fami«llueft¾a 3 Vaccine

(^'00196],. Bovine RhinoiraeheftisA ms Dkr hea-Parainllueaza 3-Respitatory Syncytial Virus Vaccine

[00197], BHoaveimaeo IRythicinao-i^raasci eueirteisli-aV Miruuslt Occiaidi¾r hVea&e- Fei nreaiaflt¾ei«a 3-Respira ry Syncytial Virus-M&an eimis

[0 198],. Bovine .Rhteoiracfteitis-Vims Diarrhea- Res itatory Syncytial. Vims Vaccine

[0 1 9] . Bovine Roiavirus-Cof onavirus Vaccine

(00200).. Bovine Virus Diarrhea Vaccine

[00201 ]. Bovine Virus Diarrhea-Respiratory Syncytial Vims Vaccine 00202], Bronchitis Vaccine

[00203] . Brucell Abortus Vaccine [0 204J . Bursa! Diseasc~ arek¾ Disease Vaccine

|CKi205]. Bursal Disease-Newcastle Disease Vaccine

[00206], Bu sa! Disease-Newcastle Disease-ilronehiiis Vaccine

[002⁽ 7] . Bursal Disease-Newcastle Disease-Bro»cfe is-!leovims Vaccine

[00208], Bursa! Disease-Newcastle Disease-Re virus Vaccine

[00209 ) . Bursal Disease-Reovinis Vaccine

1 0210] . Canary Pox Vaee ine

[002 ! ! . Canine Acfeaoviras Type 2-l¾raio:i iteri¾i$~8ord e !a Bronchise Hca Vaccine

100212], Canine Corooavi r s Vaccine

[00 13], Canine Coroaavirus-Rarvovims Vacei«e

|00214], Canine Distemper Vaccine

[00215],. Canine Bisteniper-Adenovims Type 2 Vaccine

[00 161. Canine Disiemper-Ci ronavirus-P¾rv irus Vaccine

[00217] , Canine Diste per-Hepatit i -Pami iliteiiza Vaccine

[00 18 ] , Canine Distemper- M easles Vaec ine

[0021 ] Canine Disiemper-Parvoviros Vacc ne

[00220] . Canine influenza Vaccine

[00221 ], Canine Farai nfinenza Vaccine

[00222] .

Bmnchisepliea Vaccine

[00223], Chicken. Anemia Vims Vaccine [00224] . C ή lanw dia f sittaei Vaccine

[00225]. C ccidiosis Vaccine

[00226], Distemper Vaccine

[00227] . Duck V«¾s Enteritis Vseeme

[00228], Duck Virus Hepatitis Vaccine

[00229 j . iidwardsi !k !ciakiii Vaccine

[00230] Encephalomyelitis Vaccine

[023 ! . Bnee 8ioOT ei¾i$~fif1¾esi¾¾~ West Nik Virus Vaccine

[00232],

Vaccine

[00233], Equine Arteritis Vaccine

100234], Equine influenza Vaccine

[00235 ] Equine Rhi nopneunmtiitis Vaccine

[00.236] . Equ e Rhinopie msinitis-l ilu oza Vaccne

[00237], E uine Rotavirus Vaccine

[00233] . Ery sl elothrix Rtetopai ae Vaccine

[00239], Escherichia CoH Vaccine

[00240] . Feline Calici virus Vacci ne

[00241 ], Feline immunodeficiency Virus Vaccine

[00242]. Feline lma¾ nodeficiency-Leukems¾ Virus Vaccine

[ 0024 f , Feli n e feleeiious Peri ion hi & Vaeei [00244]. Feline Leukemia Vaccine

(00245:. Feline Leukemia-Chlan dia Psitiaei Vaeclni

[00246], Feline L ukm)ia-R]iim jach ¾s^alk-Pajik kopea« Vaccine

[00247]. Feline Leukeni i a- Rb moimeneife-Ca 1 ic i- Pas I eukojtenia-Ghlm ny dsa Ps iitaei Va cine

[00:2481. Feline Pa«l ikepe»i.a Vaccine

[00249].. FeU R .imrtraeheiiis Vaccine

[00250]. Feline Rhn iwlxeiiis-Caiici-Chlamv^a Psiitari ^'Vaccine

100251; ¾l fee Rbiivotrae eitis-C lici- l¾sle«koj3 nia Vaccine

[00:252], Feine R iati^heiib alici-Pa«leuk^ni¾-Chla»iylia PsiOaes Vaccine

[00253]. Feline R!vinoiracIieiis^¾!ci~F «l iiko> iii - :lil ffiyilia t†aei«Rafcies Vaccine

[00254], Feline Rhinotrac heitis-Ca ltd- Pan i i&©pe«ia- Rabies V aeciae

[00255 ). Feline R n»ft:aehe¾is~Calicivir«s- Vaccine

[00256], Flavobacteri m Colwmaare Vacdne

(0025?;. Fowl Larvn¾oliac e,iiis Vaccine

[0O258J. Fowl i,ar fi¾otraeheiis~M .i¾k¾ Disease Vaccine

ί0025ν;. Fowl Pox Vaccine

Fowl Pox^aryngotrac eit Vaccine

[00:26 ! . F l P x- arek's Disease Vaccine

[00262]. Fowl Pox- copl sma Gal li sept scant Vaccine [00:263] . Fox Encephalitis Vaccine

[00264], Giardia Lsaa tia Vaccine

[00265], Haemophilus Parasuis Vacdne

[00266]. Hemorrhagic Enteritis Vaccine

[0026?]., Lawsoaia intracellular is Va cine

[00268). Maaaheaiiia Bamolytka-Pasieurel!a N tocida Vaccine

[00269], arek's Disease Vaccine

[00270] . Mare!cs Disease-Newcastl D sease Vaccine

(^'002? I } . Vlarek's Diseasc-Tenosynovitls Vaccine

[00272], ink Distemper Vaccine

100273], Mink Distemper-Bmeriiis- Vaccine

[00274],. Mink Enteritis Vaccine

[00:275]. Mycoplasma ClaSUseptieura Vaccine

[00276],. Neospof Cani.tiu.rn. Vaccine

[00277] . Newcastle Disease Vaccine

[00278]. ewcas le! Disease-Fowl. Pox. Vaccine

[00:279]. KewcasOe-BfOttehms Vaccine

[00280], Newcastle-Bronchitis Vaccine

[002811. NewcasOe-Par m o irus Vaccine

[00282]. Ovine Ecthyma Vaccine [002 S3]. Pacheco's i>i sesse V&cei «e

[00284] . Parainfluenza 3 Vaccine

[00285], Parvovirus Vaceine-

|IK)286| . Pasteure a Mo^'ltocida Vaccine

[00287], Pigeon Pox Vaccine

[00288). Porcine Circovirus Vaccine

[00289], Porcine l eprotl etive & Respiratory Syndrome Vaccine

[QO290I- Porcine Rotavirus Vaccin

(^'002 1 ] , Porcine Rotavmis-Tmns issihle Gastroenteritis Vaccine

[00292], Pseu rabies Vacc ne

100293], Quail Pox Vaccine

[00294], .Rabies Vaccine

[00295 ) . RiemereOa Anatipeatifer Vaccine

[00296], Salmonella Cboleraesuis Vaccine

10029/ j. Salmonella Dublin Vaccine

[00298], Salmonella Typ imnrium Vaccine

[00299]. Sifeptoeoeeas Etpi Vaccine

I00300], Swine infi«e ¾a Vaccine

[00301 ] . Tenosynovitis Vacc^'i ne

(00302). Tmnsmissibie Gastroenteritis Vaccine [00303] Trichomonas Foetus Vaccine

[0030 . Wart Vac te

[00305], West Nile Virus Vaccine

[00306] . The vaccine manufacturers referred to above are generally mnlt oational corporations operating in marry countries of the world.

[0030? ], The afore mentioned vaccine c mpo it ons or the immunogeus containe in them, may well be used in this invention in one embodinteat. Farther, vaccines agains one or more of the f m&n or animal diseases%tectious agents referred to above fern; a further embodiment of this mveniion

[0030,8 J. in accordance with another aspect of this invention there s provided an ambient temperature stable vaccine composition- comprising ironrunogen coated particles of a

pharmaceutically acceptable water soluble material, the connrosition having moisture content between about 0. i.% w/w in about 10% w/w.. The vaccine composition ma be prodoced according to the process of ibis invention as herein described.

[00300] ,. In the process aspect o f this invention, fluid containing one or more immunogens is aerosolized onto a granular particulate of a phan aeeuticaily acceptable water soluble material at a temperature from about 25 degrees Cels us to about 50 degrees Celsius preferably about 30 degrees Celsius to about 46 degrees Celsius suc that the immuoogen coats and is dried on to the particles under the aerosolizing conditions, and thereafter dried imrnmogen contesting particles having a moisture content between 0.1 % w/w to about 10% w/w are collected, giving a stabilised composition,

[003.10) . Fluid comprising one or more immunogens is preferably a suspension or dispersion of immunogens, such as viral particles, bacterial ceils or other micro organisms, or eukaryotie ceils. The fluid may he a culture medium in which, ibr example, virus particles are propagated or stored in stock. The fluid may, for example, be a culture medium or other fluid media containing the mimunogen. For example, conventional components used in the freeze dry log of bacteria and/or virus particles may be used, as are well known in the art. Examples include a mixture of sucrose, skim miitc and sterile water, or phosphate buffered media at around pH ?, containing tor example d.isodiuro, ΕΟΤΛ, egg albumin, and glycine. The ktnnatogsa containing fluid ma include one or more of amino acids, proteins, chelating agents, uffets, preservatives, stabilizers, metal antioxidants aad lubricants.

[003 I II. Preferably the immuaogen coating includes an adjovatit such as aluminium salts (alum), mummy! peptides and analogues or derivatives, saponins (for example Qulllaja saponin) or saponin containing c m ounds (for example ( SCOM#), polynucleot e or synthetic n cieie acid derivatives such as polyribonucleotides, sultur^ontaiuing compounds such as Levaonsole, polymers and heterocyclic and aromatic compounds such as Divema and p!uron e po!yols, amine and iipid-coritaining compounds, avridine, dimetfeyld cladccyl-ammoanim bromide, polyphosphate, eytoki»es(sueh as interferon} or biodegradable water in oil emulsions such as emolslfied paraffin. Other adjuvants or agents with imm nostimulatian or immuuoraodulating or antigen presenting properties, and commercial products Impran, Eraunade, Bmulsigen, and/or Amphigeu may als be used.

[00312] . Any pharmaceutically acceptable water soluble material or mixture of materials may he milked in the invention. By "water soluble" is mean I g of materia! dissolves in I ml to 1 ml of water at 20 degrees C, ^'The pharmaoeutteally acceptable water soluble materia! may comprise one or more monosaccharides, disaeeharides, polysaccharides or carbohydrates.

Examples include dextrose, msnnitoL fructose, poJyfmctossu, poiydextrose, dextrin, glucose, invert sugar. tactiiol, lactose, isorna!i maltito?, maltose, maitodexirin, sorbitol, xyiito!, sucrose., sueralo e, mauuose, galactose, xylose, arablnose, fructose,, glucosamine, galaetosann^'ne, rhamnose, 6^»0^«meihybD--gaiach>se, 2-0-acctolbeta~D-xylose_: 2 ~acets ido-2 >diox - eta-l gaIactose-4-su!phate, -acety!glueosamine, durouate, nmrumr nate,, methyl, galactnronate, galactose, arabiaose, alpha- D~manopytanose and bio olyrnefs formed by eovaleni bonding between one or more monosaccharide or disaeeharide units. Examples of car oh drates include alginate, amy lose, cellulose, earrageeRan, pectin. For convenience, monosaccharides, disaechatides, polysaccharides and carbohydrates may be collectively referred io as "sugars".

[003 ! 31 The harmace tically acceptable water soluble material may, aUernabvely, comprise a water soluble peptide or peptides (such as casein hydros e, or gelatine, or gelatine hydf soiaie), mineral sate such as . aluminum hydroxide, sodium chloride, so i m phosphate, sodium acid hosphate, EDTA s dium, magnesium chloride,, agnesium sulphate or a water soluble polymer. Water soluble polymers generally contain al least 1 mon mer units m the polymer c a n, arid form an aqueous solution in water. E amples include water soluble gums, pectin, ear oxymethvi cellulose, methyl cellulose hydroxyeth l methyleei!ulose, hydroxyethyl cellulose, hydroxyprop i methykel!ulose and irydroxypropyi cellulose.

[00314] . Water soluble pharmaceutically acceptable exelplen s, well k o n in the pharmaceutical/veterinary Held may in one em diment be til sed in this invention as the pharmaceutically acceptable water soluble material Examples of harmaceuticall acceptable excipients are provided, fo? example, in. ivlaftindale, The Ext Phar» mpoe _y 33rd Edition, The Pharmaceutical Press, London, 2002, which is Incorporated herein, by reference. Examples of pharmaceutically acceptable water soluble exoipienis Include compressible sugar, confectioner's sugar, dextraies, potassium chloride, or, sugar spheres. Two or more exelplents may be used .

[00315] . Particles of a pharmaceutically acceptable water soluble material preferably have a particle ske from 20 microns to I mm. More preferably the si¾e of the particles is from about 50 to about 20 microns.

[00316J , The process of the invention may be carried, out in any spray drying reactor, or Onidked bed spraying apparatus, as arc well known in the art , Examples include a PLC (Programmable .Logic Controller) Driven Turbojet™ fluid Bed Coater manu&ctured by BW! Hunhn (Daimiersinisse ?, D-79SS5, Steineo, (Jenitany), a PSDT Phannaceaiical Spray Dryer from Nsro, foe (Columbia, Md, 21.045 USA), and fluid bed dryers from Glatt (Ramsay, N J , 07446, USA) or Vector-tand (Marion, Iowa 52302, USA),

[0031 ?]. The rese invention is distinct from spray drying proposals known in the art in th t the hnmunogen containing fluid is sprayed onto flusdized particles and dried thereon, in contrast, m ^" s ra drying techniques a solution or slurry is sprayed i.uta an air stream and drie under the fall of gravity. The process of this invention is particularly advantageous giving rise to vaccine compositions in a .relatively short time period. For example a kg hatch of vaccine composition can be produced In less than an hour, f eeze drying of a similar amount of material may take 1 10 3 days or more. Moreover, the compositions of this in e t ion are more stable than freexe-dried vaccines.

[00318] . The fluid comprising one or mo e Inirounogens is preferabl spra ed through a noissle or spray head which delivers the sprayed fluid into the reactor, The fluid comprising one or more imrnnnogens may he sprayed into the f]nidi¾ed particles at any position from the base of the ilui femg z to the top, for example of a floidiidng bed. Spray & m s may be embedded in a iluidlzed bed or otherwise located in a reactor so as to deliver a spray of the flu d comprising one or more imrmmogens to the uidked particles 00319]. it is desirable, but sot essential to this invention to utilise fl uidizing conditions which exceed those generally used in flui bed operations in reactor. Conventionally, equipment mantrfactorers do not recommend, exceeding 50% w/v capacity of the processing chamber of fhtidked particles or materials. While f!uidked particles may comprise from

20/50% w/v capacity of the reactor, the process in accordance with an embodiment of the present invention allows tor processing weight reactor volume to be more than 50% w/v,

[00320]. in a specific, non-1 inuring, embodiment of the inven io particles may be loaded into a reactor containing a Iluidked bed, for example a spray coating apparatus which is modified to contain floidlxcd particles, such that t dkation occurs, far example, at a rate between 200 to 500 ^;: h.

[00321 ] . Fltiidi¾atio« is preferably conducted at a temperature between about 30 ^'!C. to about 46 ^*C

[00322]. A desired quantity of fluid comprising one or more imtnunogens is sprayed onto fiuidized particles of a pharmaceutically acceptable water soluble material, for example a sugar. Coaling of the particles and drying of immuoogen coating takes place in the fluidi¾ing conditions of the bed. Velocit of f I nidisaiiom and flow rate of immuoogen fluid into the fluldklog conditions are adjustable variables which allow tor the vaccine composition to be dried to a desired moisture content. The moi sture content of the vaccine composition is between about 0,1% w/ to about 10% w/w_; giving a stabilized vaccine composition. [00323] . It wsll be appreciated that reactor conditions, and flow rales of kamunogcn, including s|?.ra>½g of fluid comprising om or more «»m«»oge»s into a reactor containing, for example, a rluidked bed, may be readi y altered. Alterations may be made, for example, to f luidised air lume, liquid spraying speed, spray liquid temperature, humidity of inlet air, and the like. Where m alteration Is made to one parameter a person of getreral skill in the art to which the invention relates will readily be able to Identify any corresponding adjustments which ma be re uired In another parameter to compensate for the first sai alteration.

[00324]., Moisture content of materials is readily measured by methods known in the art. Including infrared moistore analysis such as Fourier Transfer-Near Intfared (FT/NIR) spectroscopy for example the Thermo leoSet Antarls FT/MIR aoaly¾er from Thermo Electron Corporation, altham, Mass., USA), halogen heatin moisture analyzer (for esample m M835 or 48 moisture analyser frora Olkl uS ir , Pine Brock, Ni„ 07058, USA)

[00325]. The stabilised vaccine composition preferably has particle ize from 50 to 400 microns, more preferably SO to 200 microns.

[00326] , The process according to one aspect of this in ention allows the production of a vaccine composition having a water content between about 0. 1% w/w to about 10% w/w.

Preferably, the water content is about 01 % w/w to about 4% w/w, more preferably about 0.2% w w to about 15% w/w. Freeze drying techniques produce relatively high moisture contents as a consequence of the freeze drying methodology. The high moisture content of freeze dried vaccines ma be associated with storage difficulties and loss of activity on storage. Low moisture content vaccines, such as <).. 1% w/w to about 2% w/w water content produced according to a preferred embodiment of the present invention, are parlicrdarly stable with msirstenance of activity on storage, including storage at ambient femperatwe, such as I S~3? _:( typically 25 ^*C for 30 days.

[0032? . The sisbi!ked vaccine composition according to an embodiment of thi invention comprises im.mursogen coated, particles of pharmaceutically acceptable water soluble material with the composition having moisture content between about 0.1% w/w to about .10% w/ as mentioned above. The inimnnogen coats the particles. The i munogen containing liquid used to coat the part cles may include other components including one or more of amino acids, chelating a en s, buffers, preservati ves, stahi!kets, mineral salts, antioxidants and lubricants.

[00328] . Com nents pre ent in the im unogen fl id used to er . the composition of the invention generally form part of the coating of the particles, unless evaporated during drying,

[00329 j. Preferabl the iatft ¾«oge« coating includes an adjuvant such as aluminum salts (alum), maramyl peptides and analogues or derivatives, sapo ins (lor example QuSI!aja saponin) or saponin containing compounds (for example iSCO ^iM)_< polynucleotides or synthetic nucleic acid derivatives such as polyribonucleotides, sulfur confirming compounds such as Levaroisole, polymers and heterocyclic and aromatic compounds such as Drvetn and piuronse poiyols, amine and iipid-containing compounds, avridine, dintethyldociadeeyl-ammoRinm bromide, polyphosphate, cytokines (such as interferon) or biodegradable water in oil emulsions such as emulsified paraffin. Other adjuvants or agents with imroonosiinra!ation or immunomodulating or antigen presenting properties may inc lude some of the following commercial products; !rapran, Emunade, li uisigen, or Amphigen,

[00330], The .composition of this invention is a stabilised vaccine composition. By this is meant that the vaccine composition is not inactivated on storage for 30 days at 25 X_> remaining efficacious, for example, giving rise to protective .immunity. By way of further example, a viral vaccine composition is stabilised against ioaetlvatio or stable where a drop in BID 50 {50% embryo Infective dose) on storage at 25 X for 30 days is less than one log. Preferably, the composition Is stable at ambient temperature, for periods of up to 30 days or more, such as from I to 7 days, 4 to 1 days, 7 to 30 days, or 30 to 120 days storage at ambient temperature, tor example 15-35 X, The stabil z vaccine composition ma be stored at 4deg.rees Celsius for extended periods, in contrast to traditional vaccine compositions known in the art. For example, the vaccine compositions of tins in vention may be stored for periods up to a year or more at 4 degrees Celsius. A vaccine compositio stabilised, against inaefivafion remains active In pro viding an immune response when administered to a subject whether human, animal, bird, fish or other subject in need of vaccination for protection, from disease.

[00331 J... The process of the present in vention and the vaccine composition, resulting therefrom may provide in one embodiment a live vaccine where immanogens are capable of reproduction is an immuoked host. For exam le, where the immunogens arc virus particles, the virus particles may be alive and infective at the completion of the process of the invention, and infective and st bilise ag n t motivation in e composition aspect of this invention. Live vaccine compositions of this invention may be stabilised for periods of up to 30 days or mo e at ambient tempera ures, for example 25 ^*C.

(00332). The vaccine composition, for example containing virus particles or bacteria, may he used as a carrier (such as a. vector) for delivery of DMA or RN sequences, lor example in gene therapy, or as vaccines. Many vaccines unde development as well as I» commercial production arc live attenuated viral agents which are used as vectors or a carrier fo other viral or bacterial proteins, or other antigens;, as vaccine antigens Live attenuated viral agents including vims like particles, geneticall modified or recombinant viral vector (for example recombinant vaccinia, adenoviruses, baeolovirus) arc included in this invention. These types of vaccines, besides disease prevention, may be used for cancer prevention and therapy. The viral vector vaccine may also be used tor gene therapy and drug delivery.

[00333]. Accordingly the inrmunogen ma be a viral particle or sub-unit, or bacterial cell which acts as a carrier, for example, of a. nucleic acid sequence such as a DMA or RNA sequence, In gene therapy, drug delivery, cancer treatment or other purposes. The immunogen may thus not be immunogenic as such, but rather a carrier. Hence, the term immuoogeo as used herein includes an entity capable of provoking an. immune response, and an entity which is not necessarily capable of providing an immune response, hat rather acts a carrie or delivery system of, for ex m le, DNA or RNA o protei se uences.

[00334]. The vaccine composition according to an aspect of this invention, is preferably in a free flowing from, p der like form as referred, to above. Highly concentrated vaccines may be provided.

[003351. Different vaccine powders may be blended together to give multivalent vaccines, free of problems of compati ility (as the vaccines are powders}, and with great simplicity and cost effectiveness. Hence, this invention has substantial enefits in the production of multivalent vaccines. [00336]. The vaccine c m sit on ma he readily dissolved, in a pharmaceutically acceptable diluent such as uffered saline or othex compositions suitable for administration to aa animal sneh as by way of oral admsoistrapoo, subcutaneous administration, administration as an eye drop, aerosol or nasal spray or other mode of administration of vaccines known in the ar . Alternatively the vaccine composition may he formed into capsules for administration to a subject oral 1 y. Such capsules include, for example, gelatin capsules or other standard capsules used in the pharmaceutical and veterinary fields. Still f rther, and alternatively, the vaccine composition ma he iableted, opti nally- with standard tableting excspienis and carriers as are well known in the art m another embodiment the vaccine composition may be coated, for example with an enteric coming which may protect the product from degradation in the stomach and/or to allow for sustained or slow release of the active therefrom.

[0033?]. Free flowing vaccine powders may also he administered transdermal^ tor example by line powder administration across the skin as is known in the art, for example using pressurised gases such as hydrogen or helium to move small particles across the skin, using for example Powderieci™ stems, formerly from Fowderieet Pharmaceuticals PLC, {Oxford, United. Kingdom), now owned by Chiron (Emeryville, Calif ).

[00338] . Throughout this specification, and the claims which follow, unles the context re uires otherwise, the word

[003391. comprise", or variations sueb as "comprises" or "comprising", will be understood to imply the inclusion of a stated integer or step or group of integer or steps bill not the exclusion of any other integer or step or group of Integers or steps,

(003401 The .reference to any prio art in this specification is not, and. should not be taken as an acknowledgment or any form of suggestion thai that prior art forms part of the common general knowledge.

[0034! I . Non-limiting, illustrative aspects of the present invention will now be described with reference to the following examples.

[00342]. EXAMPLE I. [00343]. A H ttlin Turbojet spray dryer is modified so as to pro de a flutdteed bed of particles for contact with a sprayed immunogen containing fluid, in particular, the Turbojet spray dryer was m dified to include spray nozzles to provide a floidi¾ed bed, and spray nozzles which spray the i m nogea containing fluid from the o tom of the processing vessel is an upward direction .

(00344). Commercially available spray dryiog apparatus spray a solution or slurry into an airstreani and allow them material to dry as It: falls by gravity. The material may be subsequently sprayed to yield agglomerates. In cont ast, in this example, particles of a pharmaceutically acceptable water soluble material are added to the spray dryer so as to provide a iluidized bed. The fiukfeed bed Is sprayed with the imrounogen containin fluid,

[00345 J . Ί ¾e dryer was operated at a temperature between about 35 and 42 ^* C

[003461. EXAMPLE 2

[00347], Sugar particles, in the term of fttaa itol or dextrose orKmydrate, were loaded into the tluktized bed of Example 1 and fiuidized with air at a temperature of 35 degrees Celsius, or 42 degrees Celsius.. The fluid air ol me was 20 em/hr, A commercially available vaccine against avian infectious bronchitis virus, H 120 was diluted Π with either;

[00348], (a) 5% sucrose, 5% slum milk, purified sterile water to 90%; or

[003491. ) a solution containing gelatin, dextran, phosphate buffer at pil 7, disodinm BDTA, mannitof egg albumin and glycine.

[00350] . The -resulting fluid containing viral particles were then sprayed onto the flu ked sugar core material at a spray rate of 12 g/min per 2 kg batch at the fluid air volume of 200 em/hr,

[00351 ], The vaccine compo ition was recovered from the iluidized bed at moisture content between 0.1 to 8% as measured using infrared moisture analysis. As an alternative measure of moisture contest, the water activity as an ersdpoint of moisture contest can be measured. [0O3S2J. Viral inactivity was than measured in chicken embryos by reconstituting the vaccine composition with an equal vo ume of mlim and injecting the vaccine composition into the chicken embryos, and thereafter determining embryo infectious <3o$e ΒΙΙ . ₍ . Viral -po enc was demonstrated for the vaccine compos ti n The vaccine composition was stable and infective after 7 days storage at ambient room temperature (25*€,) as tested by viral i activit in chicken embryos.

[00353]. The H 120 virus requires storage at -15° C. to- -20° C, a¾d is very temperature sensitive. Thus this example shows vaccine stabilization,

[003541. EXAMPLE 3

100355], The process of Example 3 was carried oat using ft uidlsed mannitol particles. sprayed with a fluid containing the B 1 0 avian infectious bronchitis vaccine mixed with an equal volume of stabilizing media (b). The resulting fluid was sprayed on to the flaidized mannitol particles. A tree flowing particulate composition was recovered having a moisture level of 2 ,51%, The process normally took about 20 to 30 minutes to complete.

[00356], The same amo t of vaccine fluid was subject to freeze dryin over a 3 day time pe iod. The end products were then tested for vaccine potency by measuring viral infeetivity in chicken embryos according to Example 2, ^'lie same potency was found in both products. This example demonstrated that the dried vaccine composition according to the invention had equal potency alter production as the freeze dried vaccine. In these test both compositions were reconstituted with saline and. then tested in the chicken embryo model. The dried composition was recovered after about 30 minutes. This was in direct contrast to lite 3 day time period required to produce the feeze dried materia! ,

[00357]. EXAMPLE 4

[00358], The potency of the vaccine compositions as measured by los of infeetivit on storage at 25 "C or 35 X- wa tested, in this example,

[00359], Vaccine compositions were prepared according to Example 3 usin either the

BI20 vaccine,, or a vaccine against avian Infectious Bursal Disease (IBO). [00360]. The vaccine compositions were ompared with equivalently freeze dried preparations.

[00361 J . in this example, vaccine potency was measured by vaccine in iecfi ity in chicken embryos,

[00362]. After 7 days storage at 35 ^s C potency of the vaccine composition of this mvention (E10s¾) had reduced by less than 1 log, i in a highly potent vaccine on storage. In contrast, the freeze dried vaccine had dropped by 3 J 2 logs for the freeze dried H I 20 virus and 1 ,5 logs for the IBD freeze dried vaccine. Stability as also tested n storage at 25 degrees Celsius, After 30 days storage at 25 degrees Celsius, the vaccine omposition of this invention for the HI 20 vaccine dropped by less than I log on storage. For the i^'BD vaccine composition there was 0,85 log reduction in potency alter 30 days storage for the vaccine composition of the invention. In contrast using freeze dried 1HD vaccine as a comparison, potency dro ped by 1 ,62 logs,

[00363] . Results are shown in FIG. I where IBHO0- VB and ISD-VB refer to the vaccine compositions of the invention, respectively containing the avian Infectious Bronchitis H120 vaccine, and avian infectious Bursal Disease vaccine stored for 30 days at 25 X. 00364), This experiment shows increased stability, as demonstrated by vaccine potency, of the vaccines according to this invention, compared to tree¾c dried vaccine production,

100365]. EXAMPLE S

[00366], Newcastle Disease f 0367 j . Vaccine C mposi lions were prepared accordin to the process of Example 3 using NC I J-'eastle disease virus ("NO", La Sola strain),

|00368) . The vaccine composition was stored at 25 degrees Celsius for 30 days, and compared to a ireeze-dtied sample of the same vims stored under identical conditions. Alter 30 days, viral activity was assessed in chicken embryos according to Example 2, The compositions of the inventio were stable and effective after 30 days, showing a recinetion in viral activity of only 0.9 logs, in contrast, freexe-dned compositions showed a decrease in viral activity of 1 ,7 logs over the same period. These results are show in FIG, 1 , FIG. 1 plots decrease in viral acti ity from initial activity ibr ( e com ositions of the inventions ( D-VB).. and ibr ifee¾e dried Newcastle disease irus NP-FD). The first asd second shaded columns of this bar graph correspond to i is experiment. 00369], Storage at 25 degrees Celsius for 30 days represents extreme co dit on for vaccine storage. Even under these conditions the vaccine compos i (loss of the invention exhibit significant viral activity stewing a decrease in mieetivity of less than 1 log over 30 days of storage at 25 degrees Celsius.

! 00370;. EXAMPLE 6

[003711 Iniinynogetsscit of the Newcastle disease vaccines of Example S was tested in chickens. Four day old SP chickens were used tor the study. Ten chickens were used for eac grou ND-VB (Newcastle Disease vaccine according to the wentioa), ND-FD (freeze dried Newcastle Disease vaccine}, and control grou which received no vaocme. The Bl¾¾ 2 g (immunising infective dose) of the freeze dried composition administered was 0.8 top higher than that of the ND-V^'B composition. Each chicke received 4 rag of designated vaccine nasal !y. Their serum samples were collected ever 7 days for serology study. Hemagglutination inhibition (HI) antibod litres in scrum were determined using 4MA«> litres of Newcastle vims (Ea Sola) and 1% chicken erythrocytes solution according to a standard micro method.

(Reference: Allen, W, PL a«d It. E Gongh, ^!,A standard L¼tiaggloiin< io« inhibition lest for Newcastle disease. ! , A comparison of macro and micr methods,^" i¾. Rec. 95: 120- 123 (1974}.}

[00372] . Controls showed negligible antibody litres sgsfnst Newcastle vim The composition of the invention and freeze dried compositions demonstrated significant specific antibody litre against Ne castle disease virus.

Claims

What is claimed is:

1 , A process for the production of 8 vaccine composition of lab le immuoogens, wherein a fluid comprising one o more imrnunogens is spra ed into a reactor containing fksidtzed particles of a plmrnmeeufieaiiy acceptable wa ter sol uble material at a temperature of about 25 degrees Celsi us to a out 50 degrees Celsius., such that he s rmmogen coats and is dried onto the particles raider the tlukfemg conditions, aad thereafter collecting f m said reactor dried immnnogen containing particles .having a mois ure content, be een about 0, 1 % w/w to s out 10 % w w so as to give stabilized vaccine composition.

2, The process according to claim I whereia the Ininr nogen comprises vims particles, bacterial cells or other microorganisms, or antigenic products thereof

3, A process according to claim 2 wherein, the inrraunogen comprises a viral or baeteriaify derived Imraunogen selected from a protein, peptide, glycoprotein, or glyeolipid, or

polysaccharide, optionally associated with a carrier, which, on i«rmo»i¾at.iori of a sahject provokes an im-mune response to the virus or bacteria f which the immunogen was derived.

4, A process according to claim I wherein the fluid comprising one or more imrnunogens is a viral vaccine or bacterial vaccine preparation mixed with a stabilizing diluent to provide a .fluid comprising viral particles or bacterial imnumogens,

5, A method of preparing a vaccine composition, the method comprising; a) spraying a f!uidked of particles with a sprayed tramonogen containing fluid; h) dry the sprayed, particles of step a at a temperature ranging from 35 degrees Celsius to 42. degrees Ceicnrs.

6 , The method of cl im 5, wherein the sprayed particl e is a suga particles selected from the group of sugars: niauurtol. or dextrosemonohydmie.,

?. A method of preparing a vaccine composition, the method comprising; a) load sugar particles into a Hyi ked bed b) fl uldi e the sugar particles with air at a temperature of 35 degrees Celsius to 42 degrees Celsius at a fluid sir volume of 200 cm.%; c) dilute a eonuuereiaifv available vaccine Π wife 5% sucros , 5% skim milk, purified sterile water to 90%; d) spray the fluid containing viral particles onto, fee fiuidked sugar core material a? a spray rare of 12 g/mm per 2 kg batch at the Ouid air volume of 200 em/hr: and e) recover the vaccine co position How. the fiuidized bed at moisture content between 0.1 to 8% as measured using infrared moisture anal sis.

8, A method of reparmg a va c na composition, the method comprising: a) load sugar particles into a fiuidized bed; b) iluidi¾e the sugar particles with air at a temperature of 35 degrees Celsius to 42 degrees Celsius at a fluid air volume of 200 cm br; c) dilute a commercially available vaccine V I with a solution containin gelatin, dextram phosphate buffer at pH 7, dlsodiuru Bi>TA, maruutof egg albumin and glycine; d) spray the fluid containing viral particles onto the fiuidteed sugar core material at a spray rate of 12 g/ruiu per 2 kg hatch at the fluid air volume of 200 cm far; and e) recover the vaccine composition from the f uidi¾ed bed at . moistu content between 0,1 to 8% as measured using infrared moisture analysis.

9, The .method of claim 7, wherein fee moisture content of the recovered vaccine composition is measured by the water activity as a» endpoint of luoisture content.

10, The method of claim 8, wherein the moisture contest of fee recovered vaccine composition is measured by the water activity as an endpoint of moisture content..