TW201008578A - Vaccine composition - Google Patents

Abstract

The present invention relates to new, advantageous DTP-based combination vaccine formulations, and concomitantly administered combination vaccine kits. Methods of administration of these vaccines and kits are also provided.

Description

201008578 VI. DESCRIPTION OF THE INVENTION: TECHNICAL FIELD OF THE INVENTION The present invention relates to novel combination vaccine formulations. In order to reduce the number of vaccinations that can protect against several pathogens, reduce the cost of administration, and increase the acceptance and vaccination rate (providing protection against several pathogens), a combination vaccine is very necessary. The antigenic competition (or interference) phenomenon listed in the literature makes the development of complex vaccines more complicated. Antigen interference means that the immune response against an antigen obtained by administration of a complex antigen is often lower than that of a specific antigen administered by a specific antigen. [Prior Art] A combination vaccine is known to prevent Bordetella pertussis and Clostridium. Tetani), Corynebacterium diphtheriae 'H-type hepatitis B virus and Haemophilus influenzae type b can be prevented as appropriate (Reference, for example, WO 93/24148 and WO 97/00697). The present invention relates to the manufacture of the most desirable multivalent vaccines to date, and the use of such vaccines for preventing or treating infections of C. stipitis, tetanus, diphtheria, sputum hepatitis virus and meningococcus, preferably It is also possible to prevent or treat infections of Haemophilus influenzae, Streptococcus pneumoniae, Hepatitis B virus and/or poliovirus, wherein the vaccine component does not significantly interfere with the immune performance of any of the components of the vaccine. SUMMARY OF THE INVENTION Therefore, the present invention is to provide a complex immunological composition which confers on a host against infection by B. pertussis, tetanus, diphtheria, hepatitis B virus, poliovirus and meningococcal infection. The protective effect of disease 143905.doc 201008578, which includes: (a) Not the entire death of C. pertussis cells (Pw), which are two or more acellular pertussis components (Pa) [preferably the latter]' ( b) tetanus toxoid (TT or T), (c) diphtheria toxoid (DT or D), (d) hepatitis B surface antigen (HepB or HB), (e) deactivated poliovirus (IPV), And (f) a carrier protein and a conjugate of one or both of the capsular polysaccharides selected from the group consisting of Y-type meningococcus (MenY) and type C meningococcus (MenC), and (g) an opportunistic vector A co-consumer of protein and type B eosin. The aforementioned immunological composition may additionally comprise two, four, or a component selected from the group consisting of type A meningococcus polysaccharide [MenA] (preferably conjugated), w-type meningococcus polysaccharide 艟 [MenW] (preferably conjugated), Salmonella typhimurium Vi polysaccharide, meningococcus (preferably B serotype) outer membrane sac, or a variety of meningococcus (preferably 8 serotype) outer membrane ( Surface exposed) proteins, and death-detoxified hepatitis B virus (HepA_, well known as HaVrixTM, the product [SmithKHne Beecham (6) ah), these do not interfere with any problems. T-Call The second main idea of the present invention is to provide various beneficial kits, potted two or four complex immunological compositions, which can be used for Cimicella, Tetanus, Diphtheria, and Liver J virus. Main = 143905.doc 201008578 = Viral and Pneumococcus pneumoniae, and the protective effects of diseases caused by meningococcal and Swallow-like infections. A first specific example of the second subject of the present invention provides a kit comprising two complex immunological compositions which confers on the host against C. sinensis, tetanus, diphtheria, and hepatitis B virus , Pediatric pruritus and Streptococcus pneumoniae, and the protective effects of diseases caused by meningococcal meningococcal and gilt bacillus infection. The first container in the set includes: ' (4) Not the entire dead B. pertussis cell (Pw), which is two or more non-cellular pertussis components (Pa) [preferably the latter], (b) tetanus toxoid (TT or T), (c) diphtheria toxoid (DT or D), (d) hepatitis B surface antigen (HepB or hb), and (e) deactivated poliovirus (ιρν), and a second The container comprises: _ (a) one or more carrier proteins and a Streptococcus pneumoniae multi-enzyme [wherein the capsular polysaccharide is preferably selected from 1, 2, 3, 4, 5, 6A, 6B, 7F, 8. Conjugates of pneumonia serotypes of 9N, 9V, l〇A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F. In another advantageous embodiment of the aforementioned kit of the present invention, the first container may additionally comprise: (f) a carrier protein and a type selected from the group consisting of Y-type meningococcus (MenY) and type C meningococcus (MenC). a conjugate of one or both of the bactericidal polysaccharides, and (g) a carrier protein and a type B hemophilic bacterium (Hib) sac. 143905.doc 201008578 a mixture of saccharides; or a second container Further comprising: (1) a carrier protein and a conjugate selected from one or both of Y-type meningococcus (MenY) and type C meningococcus (MenC) capsular polysaccharide, and (〇-vector) a conjugate of the protein and the Haemophilus B. capsulatum polysaccharide; or the first container additionally comprises (f) a carrier protein and a gamma-type meningococcus (MenY) and a type C meningitis One of the bacterium of the bacterium of the genus Mycobacterium (MenC) is co-cultured, and the second container additionally includes (b) a carrier protein and a conjugate of the type B Haemophilus influenzae (Hib) capsular polysaccharide; Or the first container additionally includes (0 carrier protein and conjugate of Haemophilus influenzae type B (Hib) capsular polysaccharide, and the second container Further comprising (b) a carrier protein and a co-plant selected from one or both of γ-type meningococcus (MenY) and type C meningococcus (MenC) capsular polysaccharide. The second subject of the present invention A second specific example provides a kit comprising two complex immunological compositions that confer immunity to a host against B. pertussis, tetanus, diphtheria g, hepatitis B virus, prion, meningitis The protective effect of the disease caused by the infection of diphtheria and Haemophilus influenzae. The first container of the kit includes: (4) Not the entire dead B. pertussis cells (pw), that is, two or more non-cellular coughs Bacillus component (pa) [preferably the latter], (b) tetanus toxoid (7T or τ), (c) diphtheria toxoid (DT or d), (4) hepatitis B surface antigen (impulse or yeah), and 0) Deactivated poliovirus (IPV), 143905.doc 201008578 and the second container includes: (4) carrier protein and selected from Y-type meningococcus (MenY) and c-type meningococcus (MenC) sac Polysaccharide - or a combination of both, and (b) a carrier Conjugates of white and Haemophilus typhimurium vesicles (mb). A third specific example of the second subject of the present invention provides a ferrule comprising three complex immunological compositions which confers resistance to the host Protection against diseases caused by C. stipitis, tetanus, diphtheria, hepatitis B virus, poliovirus and meningococcus, haemophilus bacilli and pneumococcal infection. The first container of the kit includes : (a) Not the entire dead B. pertussis cell (Pw), is two or more acellular pertussis components (Pa) [preferably the latter], (b) tetanus toxoid (TT or T), (c Diphtheria toxoid (DT or D), φ (4) hepatitis B surface antigen (HepB or hb), (e) - or more deactivated poliovirus (ιρν), and the second container includes: (a) - Or a plurality of carrier proteins and a Streptococcus pneumoniae sac polysaccharide [wherein the capsular polysaccharide is preferably selected from the group consisting of 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, l〇A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F The conjugate of pneumoconiosis type, and the third container include: 143905.doc 201008578 Ο) carrier protein and selected from γ-type cerebral dysentery (MenY) and c-type meningococcus (MenC) One or both of the bactericidal polysaccharides, and (b) a conjugate of the carrier protein and the Haemophilus influenzae vesicle (mb). Any of the foregoing kits or kits of the present invention may additionally comprise one, one, two, four, five, six or seven components selected from the group consisting of Meningococcus meningitidis [MenA] (preferably Conjugated), meningococcal polysaccharide [MenW] (preferably conjugated), Salmonella typhimurium% polysaccharide meningococcus (preferably B serotype) outer membrane cyst, one or more meninges Adhesin (preferred B serotype) outer membrane (surface exposed) protein, HepA (as described above) and one or more S. pneumoniae proteins (preferably surface exposed widely these do not substantially interfere with any The problem of the antigen in the composition. The containers of the kit may be individually packaged, but preferably packaged together. A preferred list of two or three container vaccines is provided in the kit. The inventors have surprisingly discovered The kits provided provide optimal presentation of the various antigens to the host immune system. This kit provides the benefit of one or more (preferably 2 or 3, best of all) owners of pharmaceuticals. Good way to immunize the host: All antigens have protective effects, minimal side effects after immunization, minimal carrier inhibition, minimal adjuvant/antigen interference, or minimal antigen/antigen interference. In this approach, the least number of times (twice) is administered ( It is preferred to visit the medical staff at the same time) to achieve these objectives. Although in the preferred embodiment the first and second (and third) containers 143905.doc 201008578 towel vaccine is the same-time vaccination The μ position (as described hereinafter), but in another embodiment, the present invention contemplates mixing the first and second containers (preferably in an immediate manner) as a single vaccine prior to administration. [embodiment] vaccine antigen

Preparation of tetanus toxoid (ττ) is a well-known method in the art. For example, hydrazine is preferably purified from the culture solution of tetanus bacillus, and then prepared by chemical detoxification, but the other method is to purify the toxin. Recombinant or genetically engineered detoxified analogs (for example, as described in European Patent No. 2,9281). 'Tetanus _, also having an immunological fragment of the full length protein (e.g., fragment c-reference European Patent No. 478602). Preparation of diphtheria toxoid (DT) is also a well-known method in the art. For example, DT is preferably prepared by purifying toxin from a culture solution of diphtheria bacilli, and then prepared by chemical detoxification, but another way Recombinant or genetically engineered detoxified analogs of purified toxins (e.g., U.S. Patent No. 4,709,017, U.S. Patent No. 5,843,711, U.S. Patent No. 5,601,827, and U.S. Patent No. 5,917,017). Non-cell pertussis components (Pa) are well known in the art, and examples include sputum cough toxoid (PT), filamentous hemagglutinin (FHA), pertussis protein (pR... and agglutinogens 2 and 3. These antigens For partial purification or high purification, it is preferred to use 2 or more non-cellular pertussis components in the vaccine, and more preferably 2, 3, 4 or all 5 non-cellular pertussis components in the vaccine, preferably including PT, FHA and PRN» can be used to prepare ρτ in any way. For example, the toxin is purified from the culture solution of B. pertussis, and then chemically detoxified, but 143905.doc 201008578 Another way is to purify PT by genetic engineering detoxification. (e.g., U.S. Patent No. 5,085,862) » The preparation of cells (Pw) for the entire death of C. elegans suitable for the present invention is disclosed in WO 93/24148, which is the preparation of DT_TT-Pw-HepB and DT-TT-Pa-

A suitable formulation of the HepB vaccine. Deactivated poliovirus (IPV) preferably includes the 1, 2, and 3 types of vaccine technology standards. The best is Shaq polio vaccine. The main pneumococcal vaccine of the present invention comprises a polysaccharide antigen (preferably conjugated), wherein the polysaccharide is derived from at least four selected from the group consisting of 1, 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F Pneumococcal serotypes' preferred four serotypes include 6B, 14, 19F and 23F, more Preferably, the composition comprises at least seven serotypes, for example derived from 4, 6B, 9V 'Μ, 18C, 19F and 23F serotypes, and more preferably more than seven serotypes in the composition, such as at least 丨丨A serotype, such as a composition in a specific example, includes 1 i species derived from! Cyst polysaccharides (preferably conjugated) of 3, 4, 5, 6B, 7F, 9V, 14, 18C, 19F and 23F. In a preferred embodiment of the invention, at least 13 polysaccharide antigens (preferably conjugated) are included, and the invention may additionally comprise a polysaccharide antigen, such as 23 valence (such as serum 1, 2, 3, 4, 5, 0B) , 7F, 8, 9N, 9V, l〇A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 2〇, 22F, 23F and 33F serotypes). In the case of vaccination of the elderly (for example, prevention of pneumonia), it is advantageous to include the 8 and 12F serotypes (and preferably 143905.doc -10- 201008578 = 15 and 22) in the preferred preferred 11-valent antigen composition, To form a 13/15 vaccine, and to prevent otitis media in infants or toddlers, preferably 6A and 19A serotypes to constitute a 13-valent vaccine

Co-consumer: The bacterium capsular polysaccharide conjugate comprises any vector 3, poly 3 or protein containing at least one tau-helper cell determinant. Preferably, the carrier protein used is selected from the group consisting of tetanus toxoid, diphtheria toxoid, CRM197, recombinant diphtheria toxin (eg, U.S. Patent No. 4,7G9, G17, W〇93/2521〇, WO Can or Qing Guan Guan) Any of the above), pneumonia lysin derived from pneumonia bond = bacteria (preferably detoxification by chemical means, or detoxification of mutants), Mpc derived from meningococcus, and bloodthirsty Protein D of the genus bacillus (European Patent No. 59461()). Because of the known carrier inhibiting action, it is more advantageous if each of the present invention compositions contains more than one carrier of such polysaccharide antigens (?, antigen). The carrier protein of the & type can (respectively) carry (η·1} multi-audio, and another carrier protein

A polysaccharide carrying one polysaccharide, or (η_2), is carried on one carrier protein, and two polysaccharides are carried on another carrier protein, for example, in a vaccine containing four bacterial polysaccharide conjugates. On the carrier protein.肀, 1, 2 or all of the polysaccharides may be conjugated. However, the benefit of protein D is that it can be used as a carrier protein for the composition of the invention, since it can be used in various combinations (2, 3, 4 or more) in the composition. The polysaccharide body does not have a significant carrier inhibitory effect. The optimal Hib is a ruthenium conjugate, the pneumococcal polysaccharide is a protein d dt or CRM197 (4), and MeNA, MenC, MenY & Mem^TT or PD co-host, protein D may also be a useful carrier protein for providing Another 143905.doc 201008578 An antigen against the protection of Haemophilus influenzae. The polysaccharide can be attached to the carrier protein by any known method (for example, U.S. Patent No. 4,372,945, filed on Jan. 1, s. 75 7), preferably, is accomplished by CDAP conjugation (WO 95/08348). In CDAP, the cyanating agent 1-cyano-dimethylaminoindole tetrafluoroborate (CDAP) is preferably used to synthesize a polysaccharide-protein conjugate. The cyanation reaction is carried out under relatively mild conditions to avoid hydrolysis of the alkali-sensitive polysaccharide. This synthesis can be coupled directly to the carrier protein. Characteristics of the immunological composition of the present invention The immunological composition of the present invention is preferably formulated into a vaccine which is administered to a host by in vivo administration, and the immunity of the individual components in the composition formulated by this method is substantially not affected by the other The individual components in the composition are destroyed. The term "substantially not destroyed" means that after inoculation, the antibody titer against each component is higher than 60% of the titer obtained by a single antigen, preferably 70% or more. More than 80%, better than 90%, and the best is 95-100% higher. Interestingly, if it is possible to inoculate with the aforementioned kit combination, the antibody titer against the Hib capsular polysaccharide or a pneumonia polysaccharide is close to 100%. The potency obtained by using a single antigen individually, or higher than the individual. The titer obtained by administering a single antigen. Vaccine Formulation The immunological composition of the present invention is preferably formulated into a vaccine which is administered to a host by in vivo administration, and the antibody titer imparted is superior to the human receptor and can be accepted as a percentage of 143,905.doc •12-201008578 The seroprotective effect of each antigenic component is regulated, which is a test that evaluates the effectiveness of the entire population on vaccines. Antigens that are considered by the host to convert to anti-antigen sera are well known for antigens having the above-mentioned related antibody valencies. Some organizations have published such titers, such as WHO. Preferably, more than 80% of the host statistics show that the sample is converted, preferably more than 9%, more preferably than 93%, and the best is 96-100%. * The immunological composition of the invention is preferably adjuvanted. Suitable adjuvants include mineral salts such as alumina gum (aluminum gum) or aluminum phosphate, but may also be calcium, iron or zinc beta salts, or acetaminophen tyrosine, acetylated sugars, anionic or cationic derivatization Multi-brew, or an insoluble suspension of polyphosphazenes. The adjuvant of choice may also be a preferred reactive TH1 type elicitor to aid in cellular mediator responses in the immune response. High levels of Th1-type cytokines help induce a cellular immune response against a particular antigen. High levels of Th2-type cytokines help induce a liquid immune response against the antigen.适当 A suitable adjuvant system that promotes a significant Th 1 response, including the monostearate lipid a or its derivatives 'especially 3-des-quinone-acetylated monophosphate lipid a, and the monocrack acid-based lipid A combination' The combination of 3_de_〇_acetylated monosquaric acid lipid A (3D-MPL), which is mixed with aluminum salt, is related to the combination of monophosphate lipoa and saponin derivatives (especially QS21 and 3D-MPL). The enhancement system disclosed in w〇94/00153, or a composition with lower side-effect reactivity (where QS21w cholesterol termination activity) is disclosed in WO 96/33739, with QS21, 3D-MPL and fertility dissolved in oil-in-water Alcohol-related special potential adjuvant formulations are disclosed in WO 95/17210, which may additionally include saponin, more preferably 143905.doc 201008578 is QS21. Oil-in-water emulsions and tocopherols may also be included in the formulation (WO 95/17210). Ungerminated CpG (WO 96/02555) containing a nucleotide is also a preferred inducer of the TH1 reaction and is suitable for use in the present invention. The aluminum salt is a preferred adjuvant in the aforementioned immunological composition. Specifically, the HepB should preferably be adsorbed by the aluminum phosphate before being mixed with other components, and the pertactin should preferably be oxidized before being mixed with other components. Ming adsorption. In order to reduce the level of adjuvant (especially the aluminum salt) in the combination of the invention, the polysaccharide complex is unadjuvanted. The invention also provides a method comprising the steps of mixing a vaccine component with a pharmaceutically acceptable excipient to prepare a vaccine formulation. Particularly preferred DTPa compositions of the invention (either alone or in the first container of the foregoing kit) include: TT, DT, Pa (preferably including PT, FHA and PRN - preferably PRN adsorbed by aluminum hydroxide) ), HepB (preferably squamous acid adsorption), IPV, MenC (preferably a total of protein D, TT, DT or CRM197), and optionally MenY (preferred conjugated protein D, TT, DT or CRM197) The composition also includes Hib (preferably conjugated TT and/or no adsorbed adjuvant) as appropriate. Preferred vaccines are packaged in 2 vials, the first bottle comprising the liquid DTPa-IPV-HepB, and the second bottle comprising the freeze-dried version of MenC (as appropriate, MenY and/or Hib), preferably in an anti-clotting agent ( Freeze-dried in the case of sucrose or lactose. The ingredients in the bottle can be mixed in a container prior to administration to the host for single administration/injection. The composition can also be used in the aforementioned kits (ingredients in the first container). To include one or both of Hib (preferably conjugated TT and/or unadjuvanted adsorption) and/or MenC and MenY (preferably a total of "protein D, TT, DT or 143905.doc -14- 201008578 For the purpose of kits of CRM197 and/or unadsorbed containers, the composition is preferably stored in a cold, east dry form, preferably in the presence of an anti-caking agent such as sucrose or lactose. Freeze-dried. For the purpose of the DTPa composition of the present invention comprising DTPa and Hib and/or MenC and MenY or both (individually used or as a component of the first container of the aforementioned set of __), / or the Men component consumes ττ, preferably to maintain the balance of the ττ component in the vaccine, so that the total content of ττ in a single container does not exceed a critical threshold (such as 4 〇, 45, 50, 60, 7 〇 or 8 〇 micrograms ττ In order to reduce, reduce or prevent the ττ immune interference or the carrier inhibition of the ττ polysaccharide, a preferred threshold is about 5 μg. The present inventors have found that the ratio of the polysaccharide: hydrazine in the aforementioned conjugate is reduced to 1: 〇 51 5 by weight (preferably 1: 1: 0.6-1.2, optimally about 1:1), which is advantageous in this respect. . For example, DTPa-HB-IPV-HibaiO-MenqTT) vaccine should preferably have a DTpw τ content lower than the standard value (for example, 24 μg) (preferably about one-quarter to two of the normal amount, and about half of the best). Approximately 12 micrograms of conjugated Hib, and a conjugated enthalpy to MenC content of about 5 micrograms TT, the total TT will be about w micrograms. Particularly preferred Hib/Pneumococcal polysaccharide compositions (individually used or as a component in a second container of one of the aforementioned kits) include: Hib (preferably 扼ττ and/or unadsorbed) and a variety of For example, more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or 11) pneumococcal polysaccharide conjugates (for example, the composition of the pneumococcal sputum of the present invention) The best include polysaccharides (serotypes 1, 3, 4, 5, 6Β, 7F, 9V, 14, 18C, 19F and 23F), preferably pneumococcal polysaccharides conjugated PD, DT, CRM197 or ΤΤ It can also be adsorbed without adjuvant, especially aluminum salts. In a particularly preferred embodiment of 143905.doc -15- 201008578, there is no adjuvant aluminum salt in the composition. An antigen may additionally be included in the composition of the invention (e.g., a type C meningococcus capsular polysaccharide conjugate [preferably conjugated protein D, TT, DT or CRM197 and/or unadjuvanted adsorption]), however, In another embodiment, the composition contains only Hib and a pneumococcal multi-audio vehicle, and in another specific embodiment of the other formulation,

Hib and pneumococcal polysaccharides are not conjugated to the same carrier protein (especially when CRM197 is a carrier protein). The vaccine can be contained in a container (whose composition is not liquid or freeze-dried), or two vials, the first containing Hib (preferably freeze-dried) and the second containing pneumococcal antigen (preferably liquid) ). Preferably, the lyophilized composition of the composition contains an anti-clotting agent such as sucrose or lactose. The components in the vial can be mixed in a single container immediately before administration to the host in a single administration/injection mode. Inoculation of such a formulation may result in an anti-Hib capsular polysaccharide antibody obtained when the antigen is administered close to or often over 100 A alone. potency. In a preferred embodiment, there is no significant adverse effect (for protection efficiency s) for the pneumonia polysaccharide conjugate in the composition compared to the single administration, and the last major inoculation is administered. The main geometric mean concentration (GMC) of the anti-polysaccharide antibody was determined 1 month after the main vaccination was the initial administration - usually three months in the first year of life. The vaccine of the present invention (micrograms/ml) should preferably exceed 55% of the GMC obtained by administering the pneumococcal polysaccharide without the Hib conjugate (more preferably more than 6〇, 7〇, 8〇 or 9〇%). Finally, the main drug administration method uses the vaccine of the present invention for a month after comparison with the case where the Hib-free vehicle is not used, and if the antibody concentration is not less than 〇5 μg/ml, the percentage of the receptor does not exceed 10% (preferably lower than 9, 7, 5, 3 or 1%), showing no adverse 143905.doc 201008578 effect. Although the foregoing refers to Hib 'Pneumococcal and meningococcal 'polysaccharides, the present invention can be extended to Hib and Staphylococcus, large and small multi-enzymes, and oligobiliary ' (the size of the treated polysaccharide will be reduced' There is still a protective immune response that induces the host) 'These are well known in the art of vaccination (see, for example, European Patent No. 497525), and beneficially, MenY can be 〇, 〇2, 〇3, 〇*, 〇·5, 〇. 6. The oligosaccharide conjugate of 原始.7, 〇.8 or 〇.9 times the original polysaccharide is present. Another object of the invention is to provide an immunological composition or vaccine that can be used as a remedy for use in this patent specification. Another object of the present invention is to provide the present invention for the treatment or prevention of infection by B. pertussis, tetanus, diphtheria, hepatitis B virus, poliovirus and meningococcus (and Haemophilus influenzae). The immune composition of the disease, in addition, the immunological composition of the present invention is also prepared for treating or preventing B. pertussis, tetanus, diphtheria, hepatitis B virus, poliovirus, plague bacillus, pneumonia chain A group of diseases caused by cocci and meningococcal infections. In addition, the present invention also provides immunity against diseases caused by infection of B. pertussis, tetanus, diphtheria (four), hepatitis B virus, polio phase # and meningococcus (and optionally Haemophilus). Methods, the methods comprising administering to a host an immunoprotective dose of an immunological composition of the invention. Another main finger of the present invention provides a host against the 143905.doc 17 201008578 B. pertussis, tetanus, diphtheria, hepatitis B virus, poliovirus, and one or more of the bacterium, cold pneumonia, pneumonia An immunological method for diseases caused by Staphylococcus aureus and meningococcal infection, which is a simultaneous administration schedule. ^ Simultaneous administration schedule includes a set of different lymph node positions in the host (for example, one set of the present invention) The immunoprotective dose of the immunological composition in the first container of the group), and the step of administering the immunological composition in the second (or third) container in the set at the lymph collection different from the previous one Preferably, the different positions are on different limbs, and the preferred vaccine is administered within 24 hours, and the better one is on the same day, and the best one is when the same visitor is visited by the host. Preferably, the host system administers two (or all) vaccines in one or more (preferably 2) doses in the same manner, each interval of 2-12 weeks (preferably about one month), the third vaccination The administration is usually between 2 weeks and 7 months after the second vaccination. For example, the administration method of the aforementioned vaccine can be based on the normal administration schedule of the DTP vaccine (such as three visits, one visit interval per visit, for example, age 3) 4, 5 and 5 months; or 3, 5 and 11 materials 'or 3, 5 and 12 months), this administration schedule allows immunization of antigens in two (or all) containers in the anti-set The response is the highest. It can be administered in the same way from the next year of life to the adulthood. Although it is better to use the intramuscular route to administer the vaccine, the advantage of chasing the drug is that it can be administered through the mucosa. Mucosal adjuvant may be present. (preferably laureth 9 or a thermolabic toxin [LT] derived from Escherichia coli or a mutant sputum or fragment thereof) (for example, a vaccine is easy to administer a nasal vaccine, and particularly before the host) The method of parenteral administration is 143905.doc •18·201008578 is not good, and the vaccine administration position does not need to be collected into different lymphoids. The present invention also includes the simultaneous use of the container of the present invention for preparing a vaccine kit. Use of the immunological composition. Kits comprising two or more containers of TT Another subject of the invention is a vaccine kit for simultaneous administration (as defined above) in which the ττ component in two or more containers is balanced enough to effectively reduce , reduce or prevent sputum immune interference or carrier inhibition of ττ conjugated polysaccharides. Ττ is a fairly good carrier, however it is known that its use in vaccine compositions is limited, especially when free butyl is also present. If used in excess, all antigens conjugated to ruthenium exhibit lower antibody titers, so an important topic in the art is how to use sputum in many different fields in large combinations of vaccines (eg as free antigens and as many listeners) The carrier of the bulk antigen" does not have the above disadvantages. The present inventors have found the best method for solving this problem; it uses a kit to simultaneously administer a drug (as described above), and the amount of sputum in the vaccine in the first container is no more than the occurrence of immune interference or carrier. The critical threshold for the occurrence of inhibition, and the content of ττ in the vaccine in the second device is no more than the critical threshold for the occurrence of immune interference or vector inhibition, so that the total amount of simultaneous drug administration is higher than the critical threshold, and is reduced. Immune interference (or vector inhibition) (ie, less than one vaccination component) is preferred and does not occur at all. The critical threshold can be 40, 45, 50, 60, 70 or 80 micrograms, preferably about 5 micrograms, so the highest amount of can be used is up to the amount of the number in the set of containers multiplied by the criticality. Threshold. 143905.doc • 19- 201008578 The invention therefore provides a kit comprising two (or three) containers comprising two (or three) immunological compositions for simultaneous administration, each combination

Contains a free and/or conjugated form of TT in which the TT content in each container does not exceed a critical threshold to prevent or reduce the production of TT immune interference (or vector inhibition) effects, but the total amount of TT in all containers Exceeding this critical threshold. Preferably at least one of the containers should comprise free (unrecognized) TT, most preferably a DTPa or DTPw reconstituted vaccine component. Although the free TT content is in the normal range of about 42 micrograms, another advantage of the present invention is that the content ❹ (10-30 or 10-20 micrograms, for example, 1 〇, μ, 20, 25 or 30 micrograms) can be lowered, but still It is able to elicit the best anti-ττ antibody titer with minimal (or no) immune interference or vector inhibitory effects. Preferably at least one of the containers (but possibly 2 or 3) should comprise at least one (but possibly 2, 3, 4, 5, 6, 7 or more) conjugated polysaccharide, the polysaccharide being Any of the patent specifications described herein are preferably one or more pneumococcal polysaccharides (as described above), or MenC, MenY*Hib. A preferred kit can be any of the aforementioned sets of the invention. Preferably, one, two, three or all of the polysaccharide-TT conjugates are present in the set such that the polysaccharide is reduced: TT ratio (compared to standard conjugate) to i: 0.5'1·5 by weight (Preferred 1:0.6-1.2, the best is about 1:1), this vehicle still has immune function' but helps to reduce or prevent the ττ immune interference or carrier inhibition effect. The invention further provides a method of immunizing a host with the aforementioned kit, the method comprising administering to the host at the first position of the host an immunoprotective agent in the first container of the immunoprotective agent 143905.doc •20·201008578, in the host An immunologically administrable dose of an immunoprotective dose in a second container of the host (administering an immunoprotective dose of an immunologically conjugated composition in a third container of the host at a third location in the host), wherein The first and second (and third) locations are deflated with different lymph nodes. Simultaneous administration should be carried out as described above. The first and second (and third) positions are preferably in the different limbs of the host, in the first and second (and third) containers. Preferably, the immunological composition is on the same day, preferably the host is re-administered one or more times in the same manner in the future, each interval of 2-12 weeks' more preferably twice, at intervals of about 1 to 2 months. Kits comprising two or more containers of DT or CRMI 97 Another subject of the invention is a vaccine kit for simultaneous administration (as defined above) of DT components (including DT and any of the two or more containers) Alien-free mutants such as CRM197 are balanced enough to effectively potentiate DT (or CRM197) conjugated polysaccharide antibody titers and reduce post-administration reactivity (ie, when the components in the container are injected in a single injection) It has a low reactivity when used.) DT and CRM197 are quite good carriers. However, it is known that DT-containing vaccines will produce considerable post-dosing reactivity. The inventors have found that the use of kits for simultaneous drug administration (described above) It is advantageous to have a high content (4〇_15〇 micrograms, preferably 60-120 micrograms, more preferably 70-100 micrograms, and most preferably about 95 micrograms), wherein the second container includes dt_ or CRM197 - A total of vaccines for polysaccharides are administered simultaneously. The benefits of the present invention are that a) although the DT content in the first container is high, but not sufficient to induce DT immune interference or carrier inhibition effects, b) DT- or CRM-197 polysaccharide in the first container 143905.doc 21 201008578 The body conjugate is separate, so the side effect of the vaccine in the first container will not increase, but c) the antibody titer against the conjugate to DT or CRM197 polysaccharide will not decrease, and may increase (more Those who administer conjugates have higher potency or are lower than the DT content in the first container). The invention therefore provides a kit comprising two (or three) containers comprising two (or three) immunological compositions for simultaneous administration, wherein the first container comprises a high level of DT component (DT plus Upper CRM197; preferably free or unconjugated), and the second (and third) container comprises one or more polysaccharides to a total of DT and/or CRM197. The first container preferably comprises free (unconjugated) DT, most preferably a DTPa or DTPw reconstituted vaccine. The DT/CRM197 conjugated polysaccharide may be any of the patent specifications; preferably one or more of the following: 脯杰抹益交** secret, for.

The response is comparable, preferably to increase the immune response. A preferred set of immunizations of one or more of these conjugates may be any of the aforementioned sets of the present invention.

The best is about 1:1). 143905.doc • 22· 201008578 The invention further provides a method of immunizing a host with the aforementioned kit, the method comprising administering an immunoprotective dose of an immunological composition in a first container of the host at a first position of the host, in the host second An immunologically administrated dose of an immunologically administrated dose of the host in a second container (optionally administered to the third container of the host in an immunoprotective dose of the third container), the first of which The second and third (and third) locations are deflated with different lymph nodes. Simultaneous administration should be carried out as described above. The first and second (and third) positions are preferably in the different limbs of the host, in the first and second (and second) containers. Preferably, the immunological composition is on the same day, preferably the host is re-administered in the same manner one or more times per day for 2-12 weeks, more preferably twice, at intervals of about 1-2 months. The vaccine preparation of the present invention can be administered by a systemic or mucosal route to protect or treat a mammal susceptible to infection, including by intramuscular, intra-abdominal, transdermal or subcutaneous route; or oral cavity. / Digestive tract, respiratory (eg intranasal), visceral route of mucosal administration. The antigen content selected for each vaccine dose induces an immunoprotective response without significant side effects. Such levels depend on the particular immunogen selected and how it is presented. Generally, each dose is intended to include a polysaccharide of from 1 to 10 micrograms, preferably from 0 to 50 micrograms, preferably 〇^ (7) Micrograms '1 to 5 micrograms are the best range. The vaccine protein antigen component ranges from 1-100 micrograms, preferably 5 5 micrograms, the most predominant range is 5-25 micrograms. After the initial administration of the vaccine, the subject can receive 1 or more appropriate intervals of 143905.doc -23- 201008578 to accelerate the immune function. Vaccine preparations were prepared as described in the vaccine design ("Secondary Units and Adjuvant Methods" (Powell MF & Newman MJ) (1995), Plenum Printed in New York), Fullerton described as a liposome entrapment method, USA The examples provided by the examples are for illustrative purposes and are not intended to limit the scope of the invention. Example 1: Preparation of DT-TT-Pa-IPV-HepB (DTPal PVHepB) vaccine was prepared as disclosed in WO 93/24148, marketed under the trade name Infanrix-PeNTaTM (SmithKline Beecham Biologicals) ° Example 2: Preparation of MenC Or MenC-MenY vaccine

MenC: Conjugated type C meningococcal vesicles onto protein D or TT (using CDAP technique) to contain 5 micrograms of polysaccharide per 0.5 milliliter of human dose conjugate. The pH was adjusted to 6.1 and lyophilized under sucrose.

MenCMenY: Mixing C-type meningococcal capsular polysaccharides with conjugated protein or conjugated peptone or butyl conjugated Y-type meningococcal vesicles on a total of protein D or TT (using CDAP technology) A 0.5 ml human dose contains 5 micrograms of polysaccharide per vehicle. The pH was adjusted to 6.1 and lyophilized under sucrose. Example 3: Preparation of DT-TT-Pa-IPV-HepB-MenC-MenY (DTPalPV HepB/MenCMenY) or DT-TT-Pa-IPV-HepB-MenC (DTPal PVHepB/MenC) vaccine 143905.doc -24- 201008578 . The vaccines of Examples 1 and 2 were mixed immediately (same day) before use. Example 4: Preparation of Hib-valence pneumococcal conjugate (Hib/StrepllV) vaccine In the presence of lactose, freeze-dried a total of TT to TT bacteriophage virus B capsular polysaccharide at pH 6.1 (conjugated in each dose) 10 micrograms of polysaccharides) [HiberixTM (SmithKline Beecham Biologicals)] Immediately before use (same sputum used) dissolved in 11-valent conjugated to PD pneumococcal capsular polysaccharide (1, 3, 4, 5, 6B, 7F) In the liquid solution of the 9V, 14, 18C, 19F and 23F serotypes (1 microgram of polysaccharide per vehicle in each dose), the Lung® inflammatory vaccine has been previously adsorbed by 0.5 mg of Al3+ (eg A1P04). Example 5: Clinical Trial Example 4 Vaccine Study Three doses of the German infant (3, 4, 5 months old) Example 4 vaccine and control group vaccine were administered according to the time course. The results of the immune reaction (measured one month after the last inoculation) are as follows. Anti-pneumococcal IgG antibody: GMC (micro/ml) (determined by Elisa) PS Antibody time NA group S+r%l GMC ND group s+r%i GMC anti-1 PIII 30 100 1.23 33 100 0.99 Anti-3 PIII 30 100 2.04 33 97.0 1.20 Anti-4 PIII 30 100 0.98 33 100 1.03 Anti-5 PIII 30 100 1.33 33 100 1.34 Anti-6B PIII 30 100 0.54 33 100 0.62 Anti-7F PIII 30 100 1.60 33 100 1.33 Anti-9V PIII 30 100 1.61 33 100 1.21 Anti-14 PIII 30 100 2.27 33 100 2.32 Anti-18C PIII 30 100 1.06 33 100 1.04 Anti-19F PIII 30 100 2.05 33 100 1.92 Anti-23F PIII 30 96.7 0.75 33 100 0.76 143905.doc - 25- 201008578 Group A = 1 lPn-PD + Infanrix-HeXaTM (Infanrix-Penta plus additional Hib conjugate - DTPa-HB-IPV-Hib) Concomitant (different limbs), not a combination administration. The percentage of the receptor antibody concentration is not less than 0.5 μg/ml. PS1 3 4 5 6B 7F 7V 14 18C 19F 23F D 84.8 87.9 87.9 90.9 51.5 90.9 93.9 97.0 81.8 97.0 72.7 A 86.7 96.7 76.7 90.0 50.0 93.3 90.0 90.0 80.0 96.7 66.7 Anti-PRP antibody: GMC (mg/ml) (measured by Elisa) Group D (N=34) η 21 μg/ml [%] GMC [μg/ml] Anti-PRP PIII 33 100 10.75 100% receptor has low Anti-PRP (Hib polysaccharide) antibody at 1.0 μg/ml. Approximately 6 μg/ml of GMC of Hiberix (unadsorbed Hib-TT conjugate) was administered in a similar dosing schedule. In the case of ELISA antibodies, in addition to the lower geometric mean concentration of serotypes 1, 3 and 9V found in the Iipn-PD/Hib vaccine, the immune response of infants vaccinated with 1 ipn-PD/Hib vaccine and all sera inoculated The type of iipn-PD vaccine was found to be similar, however, according to 95°/. The overlap of the confidence intervals shows that these differences are not obvious. 1 lPn-PD/Hib vaccine induces functional (adjustment of function) antibodies against all 11 serotypes. The Hib vaccine with a pneumococcal conjugate vaccine does not significantly interfere with the pneumococcal immune response, and surprisingly is more than two registered plagues 143905.doc -26· 201008578 • Miao (Infanrix-HeXa and Hiberix) It can increase the anti-prp reaction. Study Example 3 Vaccine, or Example 3 and Example 4 Vaccine Co-administration Study 1: To evaluate the safety of infantrix_peNTa in a mixed MenC conjugate vaccine by administering Hib vaccine or simultaneously administering an 11-valent pneumococcal vaccine mixed with Hiberix Sexual and immune. Both the PD and TT vectors can evaluate the MenC conjugate, and the infants can administer three doses of vaccine. When visiting medical staff, they can also be vaccinated on different limbs. 10 Study 2: Evaluation of the safety and immunity of Infanrix-PeNTa mixed with MenC-MenY consensus vaccine by administering Hib vaccine or simultaneous administration of Hiberix 11-valent pneumococcal vaccine ^ pd and TT vectors The MenC and MenY conjugates can be evaluated, and the infants are given three doses of vaccine. When visiting medical staff, they can also be vaccinated on different limbs. 143905.doc 27-

Claims (1)

201008578 . VII. Patent application scope: κ-type complex immunological composition can confer host disease against diseases caused by Bordetella pertussis, tetanus, diphtheria, hepatitis B virus, poliovirus and encephalitis. Protective effects, including: a) non-cellular pertussis components, including pertussis toxoid and FHA, b) tetanus toxoid, c) diphtheria toxoid, • d) hepatitis B surface antigen, e) detoxification polio Virus, and f) a carrier protein and a conjugate of one or both of the bacteriocysts or oligosaccharides selected from the group consisting of Y-type meningococcus red meningococcal bacterium; 2. Claim 1 An immunological composition additionally comprising - or a plurality of carrier proteins together with a bacterium vesicle multi-breast or oligosaccharide selected from the group consisting of type B haemophilus bacillus, type A meningococcus, and meningococcal meningococcus Vehicle. 3. The immunological composition of claim 1 or 2 additionally comprising a dead, attenuated hepatitis A virus. The immunological composition according to claim 1 or 2, wherein the carrier protein used is selected from the group consisting of tetanus toxoid, diphtheria toxoid, CRM197, recombinant diphtheria toxin, 〇Mpc of meningococcus, pneumonia chain Cocci pneumoniae and Haemophilus bacillus protein D. 5. The immunological composition of claim 1 or 2, which is formulated as a vaccine for administration to a host in vivo, wherein the individual components of the composition are formulated to 143905.doc 201008578 in order to prevent immunity from being The other individual components of the composition are destroyed. 6. The immunological composition of claim 1 or 2, which is formulated as a vaccine for administration to a host in vivo, which results in an acceptable percentage of a human individual having a potency superior to each antigenic component An antibody that regulates serum protection. 7. The immunological composition of claim 2 or 2, which additionally comprises an adjuvant. 8. The immunological composition of claim 7, wherein the adjuvant is an aluminum salt. A method of preparing a complex immunological composition according to any one of claims 8 to 8, which comprises the step of mixing the individual components together. 143905.doc -2- 201008578 IV. Designated representative map: (1) The representative representative of the case is: (none) (2) The symbolic symbol of the representative figure is simple: 5. If there is a chemical formula in this case, please reveal the best display. Chemical formula of the inventive feature: (none) 143905.doc