VACCINE

专利摘要:
The present invention relates to a pro-resolution mediator for its use in reducing the reactogenicity induced by the administration of a vaccine or an immunogenic composition comprising at least one antigen, and vaccines or immunogenic compositions comprising such a mediator pro-resolution.
公开号:BE1024094B1
申请号:E2016/5920
申请日:2016-12-13
公开日:2017-11-16
发明作者:Arnauld Michel Didierlaurent；Caroline Christiane Herve
申请人:Glaxosmithkline Biologicals Sa；
IPC主号:

专利说明:

VACCINE
Field of the invention
The present invention relates to the field of vaccines, particularly vaccines having reduced reactogenicity.
Context of the invention
New compositions or vaccines with improved immunogenicity are needed to address unmet medical needs. Although vaccines have beneficial protective immune responses, vaccines may also produce transient side effects, such as injection site pain, swelling and bruising, a phenomenon commonly known as reactogenicity. Although reactogenicity is transient and not considered a major safety concern, it may be a barrier to vaccine uptake within a population, and there is therefore an obvious public health benefit in reducing reactogenicity. .
Pro-resolution mediators are well known in the art and a number of candidates are in clinical trials for the treatment of, inter alia, ocular and neurodegenerative diseases. Pro-resolution mediators have been reviewed in scientific journals (see Buckley et al, "Proresolving Lipid Mediators and Mechanisms in the Resolution of Acute Inflammation", 2014, Immunity 40: 315-327, Serhan "Pro-resolving lipid mediators are leads for resolution physiology ", 2014, Nature 510: 92-101).
An object of the present invention is to improve vaccines and immunogenic compositions by reducing their reactogenicity by means of pro-resolution mediators. Summary of the invention
The present invention provides a proresolution mediator defined herein for use in reducing the reactogenicity induced by administration of a vaccine or immunogenic composition defined herein. The invention further provides a method of reducing the reactogenicity induced by administration of a vaccine or immunogenic composition defined herein, including the state of administration of a pro-resolution mediator defined in the present document. The invention also provides the use of a pro-resolution mediator defined herein for the preparation of a medicament for reducing the reactogenicity induced by the administration of a vaccine or immunogenic composition defined herein. document. The invention also provides vaccines and immunogenic compositions defined herein comprising at least one antigen defined herein and a pro-resolution mediator defined herein. The invention further provides kits comprising a pro-resolution mediator defined herein, an antigen defined herein and / or an adjuvant defined herein.
Brief description of the figures
Figure 1 - Effect of resolvin El (RvE1) on the profile of local immune cells induced by the injection of adjuvants. Figure 1 is a graph showing the number of different types of immune cells per muscle collected 24 hours after mice received different injection patterns of adjuvant and / or RVEl, as indicated on the y-axis. The names of the different types of immune cells analyzed are given next to the graph, with the corresponding color code. "Early" means total dendritic cells.
Figure 2 - Effect of 7-marinesin-1 (MaR1) on the profile of local immune cells induced by the injection of adjuvants. Figure 2 is a graph showing the number of different types of immune cells per muscle collected 4 hours or 24 hours after mice received different injection schedules of adjuvant and / or MaR1, as indicated on the axis. x's. The names of the different types of immune cells analyzed are given next to the graph, with the corresponding color code. "Early" means total dendritic cells.
Figure 3 - Effect of resolvin El (RvE1) on the profile of local cytokines induced by the injection of adjuvants. Figure 3 is a graph showing the concentration of different types of cytokines / chemokines in muscles collected after mice received different injection schedules of adjuvant and / or RvE1, as indicated on the y-axis. The names of the different types of cytokines / chemokines analyzed are given next to the graph, with the corresponding color code. Frame A provides the results obtained from the muscles collected 4 hours after the adjuvant injection. Frame B provides the results obtained from the muscles collected 24 hours after the adjuvant injection.
Figure 4 - Effect of 7-marinesin-1 (MaR1) on the profile of local cytokines induced by the injection of adjuvants. FIG. 4 is a graph showing the concentration of different types of cytokines / chemokines in a muscle collected 4 hours or 24 hours after mice received different injection schedules of adjuvant and / or MaR1, as indicated in FIG. x axis. The names of the different types of cytokines / chemokines analyzed are given next to the graph, with the corresponding color code.
Figure 5 - no effect of 7-marinesin-1 (MaR1) on specific T-cell responses induced by vaccine injection. Figure 5 is a graph showing the percentage of CD4 + or CD8 + T cells expressing at least two cytokines in mice that have received different vaccination patterns, as indicated on the x-axis. Frame A provides the results obtained when the extracted immune cells were stimulated with the HBS antigen. Frame B provides the results obtained when the extracted immune cells were stimulated with the OVA antigen.
Figure 6 - no effect of 7-marinesin-1 (MaR1) on the specific antibody-type responses induced by the injection of a vaccine. Figure 6 is a graph showing the concentration of IgG antibodies in mouse sera that have received different vaccination patterns, as indicated on the x-axis. Frame A provides the results obtained using anti-OVA antibodies. Frame B provides the results obtained using anti-HBS antibodies. detailed description
It is known that vaccines can sometimes be associated with reactogenicity. Reactogenicity refers to a subset of side effects that are associated with the inflammatory response due to vaccination. Side effects can be divided into local effects (eg, pain, swelling, erythema, and induration) and systemic effects (eg, fever, nausea / vomiting, diarrhea, headache, fatigue, and myalgia). Improving vaccines by reducing their reactogenicity can improve ease of access to vaccines by specific populations, for example by reducing adolescent pain and fever in infants. As a result, reduced reactogenicity can improve vaccine uptake, thereby covering a larger portion of the population and thus reducing morbidity / mortality. In addition, excessive inflammation may also possibly negatively affect the quality of the immune response induced by a vaccine or immunogenic composition.
An object of the present invention is therefore to reduce the reactogenicity of vaccines. Accordingly, the present invention provides a proresolution mediator defined herein for use in reducing the reactogenicity induced by administration of a vaccine or immunogenic composition defined herein.
The reactogenicity can be evaluated directly in specific in vivo models by measuring body temperature, heart rate and / or psychomotility with an implant, or indirectly by following biomarkers in the blood of an animal that indicate the occurrence of an inflammatory response that may be associated with reactogenicity (eg, CRP, PGE2). Alternative in vitro models can also be used: they are mainly based on the activation of human cells by the test formulation, which can lead to the release of molecules with pyrogenic properties (see Schindler S. et al. of pyrogen tests based on cryopreserved human primary blood cells Immunol Methods, 2006, October 20, 316 (1-2): 42-51). Resolution of the inflammatory response
Infection or injury to tissue, or vaccination, usually results in an acute inflammatory response, the triggering of which is responsible for the reactogenicity that may be associated with vaccines or immunogenic compositions. Said acute inflammatory response is typically divided into two distinct, successive phases, namely initiation and resolution. Therefore, the extent and duration of the acute inflammatory response can be regulated at two levels. On the one hand, by targeting compounds that inhibit the inflammatory response (antagonists) that will have a specific impact on the initiation phase, to limit the duration and the importance of the response. On the other hand, by targeting compounds that will specifically and actively promote the resolution of the inflammatory response (agonists or pro-resolution mediators). Pro-Resolution Mediators
For the purpose of the present invention, the term "pro-resolution mediators" is intended to mean compounds that promote the resolution of the inflammatory response, in contrast to compounds that inhibit the inflammatory response. At tissue and cell level, inflammation resolution can be broadly defined as the clearance rate of polymorphonuclear cells (PMNs) to the point where they are absent from the site of primary tissue injury and return to the site. homeostasis. Key steps in this process include: 1) clearance of "danger" stimuli; 2) the catabolism of local survival signals and the silence of intracellular proinflammatory signaling pathways; 3) standardization of chemokine gradients and apoptosis of PMNs; and 4) efferocytosis (removal of debris by macrophages, especially apoptotic neutrophils) by macrophages derived from tissues and monocytes.
Pro-resolution mediators are characterized by their ability to foster / improve any of the above steps. Some of them also have a direct effect on the reduction of pain, by acting on the receptors of the terminal nerves, and can also accelerate healing and tissue repair. The mechanisms involved in the resolution of acute inflammation are described and discussed, for example, in Buckley et al. (Proresolving Lipid Mediators and Mechanisms in the Resolution of Acute Inflammation, 2014, Immunity 40: 315-327) and Serhan ("Pro-resolving lipid mediators are leads for resolution physiology," 2014, Nature 510: 92-101).
Thus, the activity of a pro-resolution mediator can be evaluated, for example, by measuring under inflammatory conditions, for example after vaccination, (i) the infiltration profile of the local immune cells (this is ie the determination of the proportion of each type of immune cells, such as for example macrophages, neutrophils, eosinophils, NK cells, T lymphocytes and / or B cells in the infiltrate) and determining if the compound to be evaluated is capable of modulating the profile, and / or (ii) the local neutrophil apoptosis status and determining whether the compound to be evaluated is capable of increasing the number of apoptotic neutrophils (e.g. specific marker recognizing neutrophils and co-staining with annexin V or propidium iodide to discriminate apoptotic cells), and / or (iii) the profile of inflammatory cytokines and chemokines. ocals and / or the presence of resolution macrophages (for example, one can distinguish between resolution macrophages and inflammatory macrophages through specific markers) and determining whether the compound to be evaluated is able to modulate the profile. Those skilled in the art are familiar with the tests and techniques to be used to evaluate any of the above. For example, after injection of a vaccine, in the absence or presence of a pro-resolution mediator, pro-resolution activity can be monitored and evaluated at different time points after injection at the injection site, for example, by collecting muscles, (i) extracting the immune cells, staining them with specific markers and determining the content of each cell type, for example, by flow cytometry, and / or (ii) by measuring cytokine levels in clarified homogenates obtained after homogenization of the collected muscles. Such a measurement can be carried out by any conventional protein detection technique, such as, for example, an Elisa assay or ball-based immunoassays which simultaneously allow the treatment and measurement of multiple proteins within a single reaction. , commonly called multiplex immunoassays.
At the injection site of a vaccine, for example, not only proinflammatory mediators are produced, but also local mediators that are anti-inflammatory and local pro-resolution mediators are produced that mediate recovery from inflammation and pain. Therefore, anti-inflammation and pro-resolution are distinct mechanisms to fight inflammation. The present invention relates to the use of pro-resolution mediators rather than anti-inflammatory molecules, such as, for example, COX2 (cyclooxygenase-2) inhibitors. The actions of the pro-resolution mediators are in stark contrast to those of currently used anti-inflammatory therapeutics (eg, COX and LOX inhibitors), which may be resolution inhibitors. For example, while anti-inflammatory mediators block the recruitment and entry of neutrophils into the lesion site, pro-resolution mediators promote clearance of neutrophils recruited and present at the lesion site by efferocytosis. The interruption of the acute resolution process leads to uncontrolled inflammation that is implicated in the pathogenesis of many chronic diseases.
Therefore, pro-resolution mediators have been proposed for treating pain, for example postoperative pain, arthritis pain, dental pain, lower back pain and inflammatory bowel disease (see WO 11/034887). In addition, pro-resolution mediators have been proposed for use in the treatment of asthma / inflammation of the airways (WO 05/089 744) and for use in the treatment / prevention of neovascularization and hemangiogenesis (WO 09/254 873).
It has been shown that COX2 inhibitors may actually negatively affect the production of pro-resolution mediators and may reduce the immune response against the antigen. Surprisingly, however, the present inventors have demonstrated that through the use of proresolution mediators, the profile of local immune cells and the profile of local cytokines (which underlie acute inflammation and reactogenicity) can be modulated ; advantageously, the immune response directed against a vaccine / immunogenic composition, when such pro-resolution mediators are used, is not adversely affected.
A particular family of proresolution mediators are lipid-derived molecules that are derived from polyunsaturated fatty acids (PUFAs). Such lipid-derived pro-resolution mediators are known to those of skill in the art and have been reviewed in scientific journals (see Buckley et al., "Proresolving Lipid Mediators and Mechanisms in the Resolution of Acute Inflammation", 2014, Immunity 40: 315-327; Serhan "Pro-resolving lipid mediators are leads for resolution physiology", 2014, Nature 510: 92-101). Therefore, in one embodiment, the pro-resolution mediators for use in the present invention are derived from PUFA. The present invention contemplates in particular those derived from PUFA ω-3 eicosapentaenoic acid (EPA), for example E-type resolvins, or PUFA ω-3 docosahexaenoic acid (DHA), for example D-type resolvins, protectins and marines, or arachidonic acid ω-6 (AA), for example lipoxins. Therefore, in particular embodiments, the pro-resolution mediators for use in the present invention are derived from PUFA ω-3 eicosapentaenoic acid (EPA), PUFA ω-3 docosahexaenoic acid (DHA), or PUFA ω-6 arachidonic acid (AA). In particular, proresolution mediators are selected from the group consisting of: resolvin (e.g., RvE1, RvE2, RvD1), protectins (e.g., protectin D1 (PDI), also known as neuroprotectin DI (NPD1) ) when acting on the nervous system), lipoxins (e.g., lipoxin A4 (LXA4)) and marines (e.g., MaR1) or any combination of two or more thereof (eg example, RvE1, RvE2 and / or RvD1 in combination with MaR1). Resolvines can be divided into 2 types: type D which derives from DHA (for example, RvDl, RvD2, RvD3 and RvD4); type E derived from EPA (e.g., RvE1, RvE2 and RvE3). In particular embodiments, the pro-resolution mediator used in the present invention is selected from the group consisting of: RvE1, RvE2, RvE3, RvD1, RvD2, RvD3, RvD4, MaR1, PDI / NPD1, 17-HDHA and LXA4 , or a functional analogue thereof, or any combination of two or more thereof. The chemical structures of D resolvins (RvD1, RvD2, RvD3 and RvD4), E-type resolvins (RvE1, RvE2 and RvE3), protectins (PD1 / NPD1) and marines (MaR1) are disclosed, for example, in Buckley et al. (Proresolving Lipid Mediators and Mechanisms in the Resolution of Acute Inflammation, 2014, Immunity 40: 315-327) which is hereby incorporated by reference. The chemical structure of LXA4 is disclosed in Serhan et al. ("Resolving inflammation: dual anti-inflammatory and pro-resolution lipid mediators", 2008, Nat Rev. Immunol 8: 349-361) which is incorporated herein by reference.
In a particular embodiment, the pro-resolution mediator is RvE1 or a functional analog thereof. In a further particular embodiment, the pro-resolution mediator is MaR1 or a functional analog thereof.
The pro-resolution mediators used in the present invention may be of natural or synthetic origin. Synthetic mimetics may be easier to produce and may be advantageous in being chemically stable. Chemically stable mimetics of many pro-resolution mediators, including RvE1 and RvD1, are known in the art (see Serhan & Petasis Resolvins and Protectins in Inflammation-Resolution, 2011, Chem Rev. 111 (10)). 5922-5943, WO 05/089 744, WO 09/154 873 and WO 11/034 887).
By "functional analogue" is meant, within the meaning of the present invention for a given proresolution mediator, a mediator whose chemical structure is modified but which retains its ability to reduce the reactogenicity and / or to modulate the inflammation.
Vaccine antigens
The pro-resolution mediators of the invention described herein are used to reduce the reactogenicity of a vaccine or immunogenic composition. The vaccines and immunogenic compositions of the invention comprise at least one antigen. By "antigen" is meant any molecule capable of eliciting an immune response in a human or an animal. For example, an antigen may be a whole organism, a protein / polypeptide, a polysaccharide, a peptide, a nucleic acid, a protein-polysaccharide conjugate, or a hapten capable of eliciting an immune response in a human or an animal, each of these types of antigen, or any combination of two or more thereof, being specifically contemplated as a possible antigen in specific embodiments of vaccines or immunogenic compositions for use in the invention . For the purposes of the present invention, the terms "protein" and "polypeptide" are synonymous and interchangeable. The immune response can be directed against a pathogen, such as viruses, bacteria, parasites or fungi. Therefore, in certain embodiments, the antigen of vaccines or immunogenic compositions for use in the invention is derived from an organism selected from the group consisting of: viruses, bacteria, parasites and fungi, or any combination of two or more of these. Alternatively, the antigen may be a tumor associated antigen, and the vaccines or immunogenic compositions of the invention may be useful for the immunotherapeutic treatment of cancers. For the purposes of the present invention, "an organism-derived antigen" includes, in particular, the organism as a whole (whole organisms, such as a whole virus or whole bacterium), or one or more molecules derived from only from the body. The antigen may be the whole organism of natural origin, and the molecule (s), for example one or more polypeptides, from the organism may be isolated and purified from such a whole organism of natural origin. Alternatively, the antigen may be artificially produced, for example using recombinant technology or using chemical synthesis. These recombinant antigens may be in a wild-type form, i.e., their nucleotide sequence or amino acid sequence is identical to the sequence of corresponding antigens derived from the whole organism of natural origin. Alternatively, said recombinant antigens may advantageously comprise one or more mutations, i.e., their nucleotide sequence or amino acid sequence includes one or more mutations with respect to the sequence of the corresponding wild-type antigens. . Whole organisms can be live attenuated or killed / inactivated. Inactivation methods using physical and / or chemical means are known to those skilled in the art.
Virus Antigen in vaccines or immunogenic compositions for use in the invention may be derived from a virus. Therefore, in particular embodiments, the antigen is derived from a virus. In particular, the antigen may be a whole virus. The entire virus can be live attenuated or killed / inactivated. Alternatively, the antigen may be a polypeptide derived from a virus.
Suitable viruses come from the Orthomyxoviridae families, such as the influenza viruses, Paramyxoviridae, for example the respiratory syncytial virus (RSV), the mumps virus or the measles virus, Togaviridae, for example the influenza virus. rubella, Papovaviridae, for example human papillomavirus (HPV), Herpesviridae, such as herpes simplex virus (HSV), human cytomegalovirus (HCMV), Epstein-Barr virus (EBV), or varicella zoster virus (VZV), Picornaviridae, such as enteroviruses, rhinoviruses, polioviruses, Flaviviridae, such as dengue viruses or hepatitis C virus (HCV), Hepadnaviridae, for example, the hepatitis B virus (HBV), Retroviridae, for example the human immunodeficiency virus (HIV), Reoviridae, for example rotaviruses, Rhabdoviridae, for example rabies viruses, or Filoviridae, as for example v Ebola irus. In one embodiment, the antigen of the vaccines or immunogenic compositions of the invention is derived from a virus selected from the group consisting of influenza virus, RSV, HPV, measles virus, the virus. rubella, mumps virus, HCMV, VZV, dengue virus, poliovirus, HIV, HBV, Ebola and rotavirus, or any combination of two or more of them.
In a particular embodiment, the antigen is derived from HCMV. Suitably, the HCMV antigen is the glycoprotein gB, which may be devoid of the transmembrane domain (as disclosed in EP 0 802 979 B1), optionally in combination with one or more proteins pp65, IE1, pUL131, gL, gH, pUL128 and pUL130 of HCMV. Suitably, the HCMV antigen is a combination of gB, gL, gH, pUL131, pUL128 and pUL130. Alternatively, the HCMV antigen is a combination of gL, gH, pUL131, pUL128 and pUL130.
In a further particular embodiment, the antigen is derived from VZV. Suitably, the VZV antigen is the glycoprotein gE, from which the transmembrane domain can be deleted, as disclosed in EP 0 405 867 B1.
In a further particular embodiment, the antigen is derived from RSV. Suitably, the RSV antigen is a polypeptide selected from the group consisting of the fusion protein (F), the binding protein (G), the template protein (M2) and the nucleoprotein (N). Particularly suitable RSV polypeptide antigens for inclusion in vaccines or immunogenic compositions according to the invention are F-conformation polypeptides. F conformation polypeptides have already been described in both the preFusion conformation (PreF) and the postfusion conformation (PostF). Examples of F-conformation-constrained protein antigens in the pre-fusion conformation have been described in the art and are disclosed in detail, for example, in WO 09/079,796, WO 10/149 745, WO 11 / 008 974 and WO 12/158 613, each incorporated herein by reference. Likewise, conformationally constrained F-type protein antigens in the post-fusion conformation are also known in the art and can be used in the vaccines or immunogenic compositions of the invention. Examples of post-fusion constrained F-protein polypeptides are given, for example, in WO 11/008974, and in Swanson et al. (PNAS, 2011, Vol 108: 9619-9624), each incorporated herein by reference. In particular embodiments, the vaccines or immunogenic compositions for use in the present invention comprise an RSV-derived polypeptide antigen selected from the group consisting of: F protein, preF protein, N protein, and M2 protein .
In a further particular embodiment, the antigen is derived from HBV. Suitably, the antigen is the hepatitis B surface antigen (HBS).
Bacteria The antigen in vaccines or immunogenic compositions of the invention may be derived from a bacterium. Therefore, in particular embodiments, the antigen is derived from a bacterium. In additional particular embodiments, the antigen is a bacterium selected from the group consisting of: B. pertussis, S. pneumoniae and N. Meningitidis, or any combination of two or more thereof. It can be an entire bacterium and it can be killed / inactivated or live attenuated.
Particular antigens of the whole bacteria type that can be used in the present invention are Bordetella pertussis. In one embodiment, the B. pertussis antigen is the whole bacterium (Pw antigen), optionally in combination with tetanus toxoid (T) and / or diphtheria toxoid (D). In particular embodiments, the vaccines or immunogenic compositions of the invention include Pw, tetanus toxoid, and diphtheria toxoid (DTPw). The Pw antigen can be inactivated by several known methods, including mercury-free processes. Such methods may include heat inactivation, formaldehyde, glutaraldehyde, acetone-I or acetone-II (see, for example, Gupta et al., 1987, J. Biol Stand 15:87 Gupta et al., 1986, Vaccine, 4: 185). Methods for preparing the inactivated Pw antigen suitable for use in the vaccines or immunogenic compositions of the invention are disclosed in WO 93/24148, which is hereby incorporated by reference. In a particular embodiment of a vaccine or immunogenic composition comprising a Pw antigen for use in the invention, the Pw component of the composition induces a reduced reactogenicity. The reactogenicity of anti-Pw vaccines is mainly caused by a lipo-oligosaccharide ("LOS"), which is the endotoxin from the bacterial outer membrane. The lipid A portion of LOS is primarily responsible for reactogenicity. In order to produce a vaccine containing a less reactive Pw antigen (compared to "traditional" anti-Pw vaccines, e.g. produced by the inactivation procedures described above), the endotoxin may be genetically or chemically detoxified and / or extracted from the outer membrane. In particular embodiments, the B. pertussis antigen of the vaccine or immunogenic composition for use in the invention comprises a "low reactogenicity" Pw antigen in which the LOS has been genetically or chemically detoxified and / or or extract. For example, the Pw antigen may be subjected to treatment with a mixture containing an organic solvent, such as butanol, and water as described in WO 06/002502 and in Dias et al. (Human Vaccines & Immunotherapeutics, 2012, 9 (2): 339-348), which are hereby incorporated by reference. In alternative embodiments, "low reactogenicity" is achieved by obtaining the Pw antigen from a genetically engineered B. pertussis strain to produce a less toxic LOS. WO 06/065139 (hereby incorporated by reference) discloses the genetic 3-O-deacylation and detoxification of B. pertussis LOS to yield strains comprising at least partially 3-O-deacylated LOS. . The B. pertussis antigen of the vaccine or immunogenic composition of the invention may therefore be a Pw antigen derived from a B. pertussis strain which has been modified to express a lipid A modifying enzyme, such as a dice. -O-acylase. In particular, such a strain can express PagL 3-O-deacylase, as described in WO 06/065139, as well as in Geurtsen et al. (Infection and Immunity, 2006, 74 (10): 5574-5585) and in Geurtsen et al. (Microbes and Infection, 2007, 9: 1096-1103), all of which are hereby incorporated by reference. Alternatively or additionally, the strain from which the Pw antigen is derived can naturally, or because of a modification, be devoid of the ability to modify its phosphate groups of lipid A with a glucosamine, has a diglucosamine backbone of Lipid A substituted at the C-3 'position with C10-OH or C12-OH and / or expresses molecular LOS species that lack terminal heptose. Such a strain, 18-323, is disclosed in Marr et al. (The Journal of Infectious Diseases, 2010, 202 (12): 1897-1906) (incorporated herein by reference). Other particular bacterial antigens that can be used in vaccines or immunogenic compositions according to the present invention are derived from Streptococcus pneumoniae. At least one streptococcal protein and / or at least one streptococcal capsular saccharide, optionally conjugated to a carrier protein, are suitably included as antigens in the vaccines or immunogenic compositions of the invention. Suitable protein and saccharide antigens derived from Streptococcus pneumoniae are described in WO 14/060385 (hereby incorporated by reference). In some embodiments, the at least one Streptococcus pneumoniae protein is selected from the group consisting of the family of polyhistidine triads (PhtX), the family of choline binding proteins (CbpX), the truncated CbpX, the LytX family (autolytic enzyme), truncated LytX, truncated CbpX-LytX chimeric proteins, PcpA (choline-binding pneumococcal A protein), PspA (pneumococcal A surface protein), PsaA (pneumococcal A protein) surface adhesion), Spl28 (Streptococcus pneumoniae 128), SplO1 (Streptococcus pneumoniae 101), Spl30 (Streptococcus pneumoniae 130), SP125 (Streptococcus pneumoniae 125) and SP133 (Streptococcus pneumoniae 133). In additional embodiments, the vaccines or immunogenic compositions for use according to the invention comprise 1 or more than 1 (eg, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16 , 17, 18, 19, 20, 21, 22 or 23) capsular saccharide
Streptococcus pneumoniae, optionally conjugated to a carrier protein. In particular embodiments, the capsular saccharide (s) of Streptococcus pneumoniae, optionally conjugated to a carrier protein, included in the vaccines or immunogenic compositions of the invention, comprise saccharides derived from serotypes selected from the following serotypes: , 2, 3, 4, 5, 6A, 6B, 7F, 8, 9N, 9V, 1 OA, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F. For example, a 7-valent vaccine or immunogenic composition may include serotypes 4, 6B, 9V, 14, 18C, 19F and 23F saccharides. A 10-valent vaccine or immunogenic composition may comprise saccharides derived from the same 7 serotypes and further include serotype 1, 5 and 7F saccharides. A 12-valent vaccine or immunogenic composition may comprise saccharides derived from the same serotypes and further include saccharides of serotypes 6A and 19A. A 13-valent vaccine or immunogenic composition may comprise the same 12 serotypes and further comprise serotype 3. A vaccine or a 15-valent immunogenic composition may comprise saccharides derived from the same 13 serotypes and further include serotype-derived saccharides. 22F and 33F. Other saccharide antigens, for example a 23-valent (such as serotypes 1, 2, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F) are also contemplated as antigens in vaccines or immunogenic compositions for use in the invention. The term "saccharide" may indicate a polysaccharide or oligosaccharide and includes both. Polysaccharides can be isolated from bacteria and can be sized to a certain extent by known methods (see, for example, EP 0 497 524 B1 and EP 0 497 525, incorporated herein by reference). reference) and optionally by microfluidization. The polysaccharides may be sized to reduce the viscosity in the polysaccharide samples and / or to improve the filterability of the conjugates. The term "conjugate" refers to a capsular saccharide covalently linked to a carrier protein. The carrier protein can be any peptide or protein. Suitable carrier proteins are described in WO 14/060385 (hereby incorporated by reference). The carrier protein may be tetanus toxoid (TT), tetanus toxoid fragment C, non-toxic mutants of tetanus toxoid, diphtheria toxoid (DT), CRM197, other non-toxic mutants of diphtheria toxoid, such as CRM176, CRM228, CRM 45; CRM 9, CRM 45, CRM102, CRM103 and CRM107 (where CRM stands for cross-reactive material), pneumococcal pneumolysin, OMPC (outer membrane protein C), heat shock proteins, pertussis proteins, cytokines, lymphokines , growth factors or hormones, artificial proteins comprising multiple human CD4 + T cell epitopes from various pathogen-derived antigens, such as N19 protein, surface pneumococcal PspA protein, iron absorption proteins, toxin A or B of C. difficile, an H. influenzae protein, pneumococcal PhtA (polyhistidine triad protein A), pneumococcal PhtD (polyhistidine triad protein D), pneumococcal PhtB (triad protein B), polyhistidine), or PhtE (polyhistidine triad E protein). In one embodiment, the at least one capsular saccharide conjugate of Streptococcus pneumoniae is conjugated to a carrier protein selected from the group consisting of tetanus toxoid (TT), TT fragment C, diphtheria toxoid, CRM197 (cross-reactive material 197), detoxified pneumolysin, protein D (from H. influenzae), PhtD, PhtDE and N19. The saccharide may be bound to the carrier protein by any known method. Other particular bacterial antigens that can be used in the present invention are derived from Neisseria meningitidis. In some embodiments, the antigen of vaccines or immunogenic compositions for use in the invention is a capsular saccharide of N. meningitidis derived from a serogroup selected from the group consisting of: serogroup A (MenA), serogroup C (MenC), serogroup Y (MenY), and serogroup W-135 (MenW), or any combination of two or more thereof, optionally conjugated to a carrier protein. Indeed, these saccharides can be suitably conjugated to any of the carrier proteins described above in connection with streptococcal saccharides. In some embodiments, the vaccine or immunogenic compositions of the invention comprise a capsular saccharide of N. meningitidis serogroup A (MenA), a capsular saccharide of N. meningitidis serogroup C (MenC), a capsular saccharide of N serogroup Y meningitidis (MenY), and a N. meningitidis serotype W-135 (MenW) capsular saccharide, optionally conjugated to the carrier CRM197 protein or the carrier TT protein. Other particular bacterial antigens derived from Neisseria meningitidis that can be used in the present invention are derived from N. meningitidis serogroup B ("MenB"). Antigens suitable for eliciting anti-MenB responses include polypeptides, lipo-oligosaccharides and / or membrane vesicles. The vaccines or immunogenic compositions of the invention may comprise one or more serogroup B meningococcal polypeptide antigens. In some embodiments, the antigen is a serogroup B N. meningitidis polypeptide selected from the group consisting of: NadA (also known as "961" protein), NHBA protein (also known as "287" protein), fHBP protein (also known as "741" protein), GNA1030 protein ( also known as "953" protein), and GNA2091 protein (also known as "936" protein), or any combination of two or more thereof, optionally in combination with an OMV derived from N. meningitidis serogroup B. These antigens will usually be present as purified polypeptides, for example recombinant polypeptides. Suitable forms of these antigens are disclosed in WO 04/032958, incorporated herein by reference. The five antigens may be present in the composition as five separate proteins, or conveniently at least two of the antigens are expressed as a single polypeptide chain (a "hybrid protein"), for example so that the five antigens form less than five polypeptides, as described in WO 04/032958. In certain embodiments, the vaccines or immunogenic compositions of the invention comprise at least the NadA protein, the NHBA protein, the fHBP protein, the GNA1030 protein and GNA2091 protein. In particular embodiments, the vaccines or immunogenic compositions of the invention comprise SEQ ID NO: 2, SEQ ID NO: 7 and SEQ ID NO: 8, as disclosed in WO 04/032958, incorporated in the present invention. present for reference. In particular additional embodiments, such vaccines or immunogenic compositions of the invention further comprise an OMV derived from N. meningitidis serogroup B as described below. Other particular bacterial antigens are the outer membrane vesicles (OMVs). These include any proteo-liposomal vesicle obtained by the dislocation of an outer membrane or the formation of vesicles from an outer membrane that comprise protein components of the outer membrane. Gram-negative bacteria, such as Neisseria, secrete OMVs during active growth. The main immunogenic components of OMVs are outer membrane proteins (OMP) and membrane bound lipopolysaccharides (LPS). OMVs can be prepared from any Gram-negative bacterium, including pathogenic neisserial bacteria, such as Neisseria gonorrhoea and Neisseria meningitidis. The OMV approach is particularly useful for serogroup B Neisseria meningitidis because its polysaccharide capsule is poorly immunogenic. Therefore, in certain embodiments, the vaccines or immunogenic compositions of the invention comprise an OMV derived from a strain of N. meningitidis serogroup B, optionally in combination with any of the serogroup B meningococcal polypeptide antigens. described above. OMVs are artificially prepared from bacteria, and can be prepared using detergent treatment (e.g., with deoxycholate), or by non-detergent means as described in WO 12/020. 326, for example, which is incorporated herein by reference.
Parasites The antigen in vaccines or immunogenic compositions of the invention may be derived from parasites. Suitably, the antigen can be derived from malaria causing parasites. Therefore, in some embodiments, the antigen in vaccines or immunogenic compositions for use in the invention is derived from malaria causing parasites, such as, for example, Plasmodium falciparum or Plasmodium vivax. Suitably, the antigen derived from Plasmodium falciparum is RTS, S. As disclosed in WO 93/10152 (hereby incorporated by reference), RTS, S is a hybrid protein consisting of the C-terminal portion of the Plasmodium falciparum circumsporozoite (CS) protein bound by the amino acid intermediate from the preS2 portion of the hepatitis B surface antigen to the hepatitis B virus surface antigen (S).
Tumor-Associated Antigens The antigen in the vaccines or immunogenic compositions of the invention may be a tumor-associated antigen. Suitably, the antigen may be a tumor rejection antigen, such as those for prostate, breast, colorectal, lung, pancreatic, renal and melanoma cancers. Non-limiting examples of antigens include MAGE 1, 3 and MAGE 4 or other MAGE antigens, as disclosed in WO 99/40188.
RNA nucleic acid immunization with self-replication
Nucleic acid immunization can be accomplished by administering self-replicating RNA (or self-amplifying RNA) encapsulated in and / or absorbed on a small particle. The RNA encodes a polypeptide antigen of interest, and the particle can deliver that RNA by mimicking the distribution function of a naturally occurring virus. After the in vivo administration of the particles, the RNA is released from the particles and is translated inside a cell to provide the antigen in situ. Any of the polypeptide antigens described above, suitable for inclusion in vaccines or immunogenic compositions according to the invention, may be expressed as a self-replicating RNA molecule encoding said antigen, as disclosed in WO 12/006376 which is incorporated herein by reference. Therefore, in particular embodiments where the antigens in the vaccines or immunogenic compositions for use in the invention are polypeptides, such polypeptides are encoded by self-replicating RNA. In such a situation, said self-replicating RNA is appropriately coupled to a delivery system, particularly lipid based delivery systems, such as a cationic nanoemulsion (CNE), or a liposome. Suitably, when the lipid-based system is a CNE, the self-replicating RNA is absorbed on the outer surface of the CNE, whereas when said lipid-based system is a liposome, the RNA is self-replicating. -amplification is encapsulated in the liposome.
By "self-replicating RNA molecule" (or "self-amplifying RNA"), it is meant that, when administered to a vertebrate cell, even without any protein, the molecule leads to the production of multiple RNA son by transcription from itself, as explained in WO 12/006 376, which ultimately results in the expression of the encoded antigen that becomes a major cell polypeptide.
One suitable system for obtaining self-replication in this manner is to use an alphavirus-based replicon, as described in detail in WO 12/006376. Suitably, said replicon encodes for (i) RNA-dependent RNA polymerase which can transcribe an RNA from the self-replicating RNA molecule and (ii) an antigen of interest. The polymerase may be an alphavirus replicase, comprising for example one or more alphavirus nsP1, nsP2, nsP3 and nsP4 proteins. Appropriate characteristics of self-replicating RNA molecules and methods for their preparation are also described in WO12 / 006,376.
In some embodiments, the vaccines or immunogenic compositions for use in the present invention comprise a self-replicating liposome and RNA encoding any of the polypeptide antigens described herein in the liposome encapsulated. In additional embodiments, the vaccines or immunogenic compositions for use in the present invention comprise a CNE and a self-replicating RNA encoding any of the polypeptide antigens described herein absorbed on the outer surface. of the CNE. In particular embodiments, the self-replicating RNA molecule encodes polypeptide antigens derived from the group consisting of: HCMV, RSV and HIV.
Examples of CNEs that can be used in the present invention, as well as methods for their preparation, are described in WO 12/006 380 which is incorporated herein by reference.
Various amphiphilic lipids can form bilayers in an aqueous environment to encapsulate an aqueous core containing RNA in the form of a liposome. These lipids may have an anionic, cationic or zwitterionic hydrophilic headgroup. Some phospholipids are anionic, while others are zwitterionic and others are cationic. Suitable classes of phospholipids include, but are not limited to, phosphatidyl ethanolamines, phosphatidylcholines, phosphatidylserines, and phosphatidylglycerols. Useful cationic lipids include, but are not limited to, dioleoyl trimethylammonium propane (DOTAP), 1,2-distearyloxy-N, N-dimethyl-3-aminopropane (DSDMA), 1,2-dioleyloxy-N, N-dimethyl-3-aminopropane (DODMA), 1,2-dilinoleyloxy-N, N-dimethyl-3-aminopropane (DLinDMA), 1,2-dilinolenyloxy-N, N-dimethyl-3-aminopropane ( DLenDMA). Other useful cationic lipids are disclosed in WO 15/095340, for example the lipids claimed in any one of claims 1 to 8 of WO 15/095340 herein incorporated by reference. Zwitterionic lipids include, but are not limited to, zwitterionic lipids of the acyl type and zwitterionic lipids of the ether type. Examples of useful zwitterionic lipids are DPPC, DOPC and dodecylphosphocholine. The liposomal particles of the invention may be formed from a single lipid or from a lipid mixture. A mixture may comprise (i) a mixture of anionic lipids, (ii) a mixture of cationic lipids, (iii) a mixture of zwitterionic lipids, (iv) a mixture of anionic lipids and cationic lipids, (v) a mixture of anionic lipids and zwitterionic lipids, (vi) a mixture of zwitterionic lipids and cationic lipids or (vii) a mixture of anionic lipids, cationic lipids and zwitterionic lipids. When a lipid mixture is used, it is not necessary that all the lipid components of the mixture be amphiphilic, for example one or more amphiphilic lipids may be mixed with cholesterol. The hydrophilic portion of a lipid may be PEGylated (i.e., modified by the covalent attachment of a polyethylene glycol). This modification can increase stability and prevent non-specific adsorption of liposomes. Liposomal particles are usually divided into three groups: multilamellar vesicles (MLVs); small unilamellar vesicles (SUVs); and large unilamellar vesicles (LUVs). MLVs have multiple bilayers in each vesicle, forming several separate aqueous compartments. SUVs and LUVs have a single bilayer encapsulating an aqueous nucleus; SUVs generally have a U 50 nm diameter and LUVs have a diameter> 50 nm. The liposomal particles useful in this aspect of the invention are ideally LUVs having a diameter in the range of 50 to 220 nm. Suitable liposome preparation techniques are well known in the art. A useful method is described in Jeffs et al. (Pharmaceutical Research, 2005, 22 (3): 362-372) and involves mixing (i) an ethanolic solution of the lipids, (ii) an aqueous solution of the nucleic acid and (iii) a buffer, followed by mixing, equilibration, dilution and purification.
Viral vectors
Alternatively, nucleic acid immunization can be performed using a vector capable of replicating or unable to replicate, such as a viral vector. Many suitable viral vectors for introducing nucleic acids encoding antigens of interest into a subject are known in the art, and include both DNA and RNA viruses. Suitable examples are: adenoviral vectors (capable of replicating or unable to replicate), poxvirus-based vectors, especially vaccinia-based vectors such as modified vaccinia virus (MVA) , NYVAC, avipox-based vectors, canarypox virus (ALVAC) and avian pox virus (FPV), alphavirus-based vectors (such as Sindbis virus, Semliki Forest, Ross River Virus, and Venezuelan Equine Encephalitis Virus) and their chimeras and replicons, herpesvirus-based vectors (eg, cytomegalovirus-derived vectors) ), arenavirus-based vectors, such as lymphocytic choriomeningitis virus (LCMV), a retrovirus, a lentivirus, pseudo-viral particles, and many others. In one embodiment, the polypeptide antigen in vaccines or immunogenic compositions for use in the present invention is encoded by an adenoviral vector. In particular embodiments, the polypeptide antigen encoded by an adenoviral vector is derived from HIV, malaria, Ebola, or RSV. The production and use of adenoviral vectors are well known to those skilled in the art. Suitable examples of description of the design, production and use of adenoviral vectors can be found, for example, in WO 05/071 093 and WO 10/086 189, which are incorporated herein by reference. reference. The adenoviral vectors that can be used in the present invention can come from various mammalian hosts. The adenoviral vectors can come from a human adenovirus. Examples of such human adenoviruses are Ad1, Ad2, Ad4, Ad5, Ad6, Ad11, Ad24, Ad34, Ad35, in particular Ad5, Ad11 and Ad35.
Alternatively, the adenoviral vectors may be derived from a non-human primate adenovirus, for example a chimpanzee adenovirus, such as one selected from the ChAd3, ChAd63, ChAd83, ChAd15, Pan5, Pan6, Pan7 and Pan9 serotypes. More specifically, the virus may be a non-human adenovirus, such as a simian adenovirus and, in particular, a chimpanzee adenovirus, such as ChAdl55, Pan 5, 6, 7 or 9. Examples of such strains are given in WO 03 / 000 283, which is hereby incorporated by reference, and is available from the American Type Culture Collection, 10801 University Boulevard, Manassas, Virginia 20110-2209, and other sources. Desirable strains of chimpanzee adenoviruses include Pan 5 [ATCC VR-591], Pan 6 [ATCC VR-592] and Pan 7 [ATCC VR-593].
The adenoviral vectors that can be used in the present invention can come from an adenovirus incapable of replicating, for example, comprising a functional deletion El. The adenoviral vectors that can be used in the present invention include PanAd3 (WO 10/086189) and ChAd155 (GB 1,510,357.5). In some embodiments, vaccine antigen or immunogenic compositions for use in the invention are recombinantly expressed in the adenoviral vector ChAd155. In particular embodiments, the ChAd15 adenoviral vector encodes at least one respiratory syncytial virus (RSV) derived antigen, particularly any of the RSV polypeptide antigens described above. The adenoviral vectors can be produced by any suitable cell line in which the virus is able to replicate. Without limitation, such a cell line may be selected from HeLa cells [ATCC accession number CCL 2], A549 [ATCC accession number CCL 185], HEK 293, KB [CCL 17], Detroit [by for example, Detroit 510, CCL 72] and WI-38 [CCL 75], among others.
admixtures
The vaccines and immunogenic compositions of the invention may also comprise an adjuvant in addition to the antigen. Adjuvants are used in vaccines to enhance and modulate the immune response directed against the antigen. However, the adjuvants may induce increased reactogenicity, and in these particular embodiments, the vaccines and immunogenic compositions of the invention comprise an adjuvant. The adjuvants described below herein may be combined with any of the antigens described hereinabove. The adjuvant may be any adjuvant known to those skilled in the art, but adjuvants include (but are not limited to) oil-in-water emulsions (e.g., MF59 or AS03), liposomes, saponins, TLR2 agonists, TLR3 agonists, TLR4 agonists, TLR5 agonists, TLR6 agonists, TLR7 agonists, TLR8 agonists, TLR9 agonists, aluminum salts, nanoparticles , microparticles, ISCOMs, calcium fluoride and composites of organic compounds or combinations thereof.
Oil-in-water emulsions
In one embodiment of the present invention, there is provided a vaccine or immunogenic composition for use in the invention comprising an oil-in-water emulsion. The oil-in-water emulsions of the present invention comprise a metabolizable oil and an emulsifying agent. For any oil-in-water composition to be suitable for human administration, the oily phase of the emulsion system must include a metabolizable oil. The meaning of the term metabolizable oil is well known in the art. Metabolizable can be defined as "being able to be metabolized" (Dorland's Illustrated Medical Dictionary, W. B. Sanders Company, 25th Edition, 1974). A particularly suitable metabolizable oil is squalene. Squalene (2,6,10,15,19,23-hexamethyl-2,6,10,14,18,22-tetracosahexaene) is an unsaturated oil that is found in large quantities in the liver oil. shark, and in smaller amounts in olive oil, wheat germ oil, rice bran oil and yeast, and is a particularly preferred oil for use in an oil-in-oil emulsion. water of the invention. Squalene is a metabolizable oil because it is an intermediate in the biosynthesis of cholesterol (Merck Index, 10th Edition, entry No. 8619). In certain embodiments, wherein the vaccines or immunogenic compositions of the invention comprise an oil-in-water emulsion, the metabolizable oil is present in the vaccine or immunogenic composition in an amount of from 0.5% to 10% (by volume / volume) of the total volume of the composition. The oil-in-water emulsion further comprises an emulsifying agent. Suitably, the emulsifier may be polyoxyethylenesorbinate monooleate (POLYSORBATE 80). In addition, said emulsifying agent is suitably present in the vaccine or immunogenic composition in an amount of 0.125 to 4% (v / v) of the total volume of the composition. The oil-in-water emulsion may optionally include a tocol. Tocols are well known in the art and are described in EP 0 382 271 B1. Suitably, the tocol may be alpha-tocopherol or a derivative thereof, such as alpha-tocopherol succinate (also known as vitamin E succinate). Said tocol is suitably present in the adjuvant composition in an amount of 0.25% to 10% (v / v) of the total volume of the immunogenic composition. The oil-in-water emulsion may also optionally comprise sorbitan trioleate (SPAN 85).
The process for producing the oil-in-water emulsions is well known to those skilled in the art. Generally, the process comprises mixing the oily phase (optionally comprising a tocol) with a surfactant, such as a solution of PBS / TWEEN8 0 ™, followed by homogenization using a homogenizer, it will be It will be apparent to those skilled in the art that a process comprising passing the mixture through the needle of a syringe twice is suitable for homogenizing small volumes of liquid. Similarly, the process of emulsification in a microfluidizer (M110S Microfluidics machine, maximum of 50 passes, for a period of 2 minutes at a maximum inlet pressure of 6 bar (output pressure of about 850 bar) can be adapted by a person skilled in the art to produce smaller or larger volumes of emulsion, adaptation can be accomplished by routine experimentation including measurement of the resulting emulsion until a preparation having droplets of oil of required diameter.
In an oil-in-water emulsion, the oil and the emulsifier must be in an aqueous carrier. The aqueous carrier may be, for example, phosphate buffered saline or citrate solution.
In particular, the oil-in-water emulsion systems used in the present invention have a small submicron oil droplet size. Suitably, the droplet sizes will be in the range of from 120 to 750 nm, more preferably diameters of from 120 to 600 nm. Even more particularly, the oil-in-water emulsion contains oil droplets of which at least 70% in terms of intensity have a diameter of less than 500 nm, more particularly at least 80% in terms of intensity have a diameter less than 300 nm, more particularly at least 90% in terms of intensity have a diameter in the range of 120 to 200 nm.
The size of the oil droplets, i.e. the diameter, according to the present invention is given in terms of intensity. There are several ways to determine the diameter of the oil droplet size by intensity. Intensity is measured using a sizing instrument, suitably by dynamic light scattering, such as Malvern Zetasizer 4000 or, preferably, Malvern Zetasizer 3000HS. A first possibility is to determine the mean diameter z (ZAD) by dynamic light scattering (photon correlation spectroscopy-PCS); this method gives in addition the polydispersity index (PDI), and both the ZAD and the PDI are computed with the cumulants algorithm. It is not necessary to know the refractive index of the particles to obtain these values. A second way is to calculate the diameter of the oil droplet by determining the size distribution of all the particles using another algorithm, the Contin, or NNLS, or the automatic "Malvern" algorithm (the algorithm by defect provided by the sizing instrument). Most of the time, since the refractive index of the particles of a complex composition is unknown, only the distribution of the intensities is taken into account, and if necessary the intensity means coming from this distribution.
ISCOMs
In some embodiments of the present invention, vaccines or immunogenic compositions of the invention comprising ISCOMs are provided. ISCOMs are well known in the art (see Kersten & Crommelin, 1995, Biochimica and Biophysica Acta 1241: 117-138). ISCOMs include saponin, cholesterol and phospholipids and form an open-cage structure generally of 40 nm size. ISCOMs result from the interaction between saponins, cholesterol and other phospholipids. A typical reaction mixture for the preparation of an ISCOM is 5 mg / ml saponin and 1 mg / ml for each of cholesterol and phospholipid. Phospholipids suitable for use in ISCOMs include, but are not limited to, phosphocholine (didecanoyl-L-phosphatidylcholine [DDPC], dilauroylphosphatidyl-choline [DLPC], dimyristoylphosphatidylcholine [DMPC], dipalmitoyl phosphatidylcholine [DPPC], distearoyl phosphatidylcholine [DSPC], dioleoyl phosphatidylcholine [DOPC], 1-palmitoyl, 2-oleoylphosphatidylcholine [POPC], Dielaidoyl phosphatidylcholine [DEPC]), a phosphoglycerol (1,2-dimyristoyl-sn-glycero-3-phosphoglycerol [DMPG], 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol [DPPG], 1,2-distearoyl-sn-glycero-3-phosphoglycerol [DSPG], 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol [ POPG]), a phosphatidic acid (1,2-dimyristoyl-sn-glycero-3-phosphatidic acid [DMPA], dipalmitoyl phosphatidic acid [DPPA], distearoylphosphatidic acid [DSPA]), a phosphoethanolamine (1,2-dimyristoyl) -sn-glycero-3-phosphoethanolamine [DMPE], 1,2-dipalmitoyl-sn-glycero-3-phosphoethane thanolamine [DPPE], 1,2-distearoyl-sn-glycero-3-phosphoethanolamine DSPE 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine [DOPE]), a phoshoserine, a polyethylene glycol [PEG] a phospholipid (mPEG phospholipid, polyglycerin phospholipid, functionalized phospholipid, activated-end phosholipid). In particular embodiments, ISCOMs include 1-palmitoyl-2-oleoyl-glycero-3-phosphoethanolamine. In additional particular embodiments, a highly purified phosphatidylcholine is used and may be selected from the group consisting of: phosphatidylcholine (egg), hydrogenated phosphatidylcholine (egg) phosphatidylcholine (soybean), hydrogenated phosphatidylcholine (of soy). In additional particular embodiments, the ISCOMs include phosphatidylethanolamine [POPE] or a derivative thereof. A number of saponins are suitable for use in ISCOMs. The adjuvant and haemolytic activity of individual saponins has been extensively studied in the art. For example, Quil A (derived from the bark of the Quillaja Saponaria Molina tree of South America), and its fractions, are described in US 5,057,540 and in "Saponins as vaccine adjuvants", Kensil, CR, Crit Rev Cher Drug Carrier Syst, 1996, 12 (1-2): 1-55; and EP 0 362 279 B1. ISCOMs comprising Quil A fractions have been used in the preparation of vaccines (EP 0 109 942 B1). These structures have been reported to have adjuvant activity (EP 0 109 942 B1, WO 96/11711). Quil A moieties, Quil A derivatives and / or combinations thereof are suitable saponin preparations for use in ISCOMs. The hemolytic saponins QS21 and QS17 (HPLC purified Quil A fractions) have been described as potent adjuvants, and their production method is described in US 5,057,540 and EP 0 362 279 B1. It is also described in US Pat. these references use QS7 (a non-hemolytic fraction of Quil A) that acts as a potent adjuvant for systemic vaccines. The use of QS21 is further described in Kensil et al. (1991. J. Immunology vol 146, 431-437). Combinations of QS21 and polysorbate or cyclodextrin are also known (WO 99/10008). Particulate adjuvant systems comprising QuilA moieties, such as QS21 and QS7, are described in WO 96/33739 and WO 96/11711, and these are incorporated herein. Other particular QuilA moieties designated QH-A, QH-B, QH-C, and a mixture of QH-A and QH-C designated QH-703 are disclosed in WO 96/011711 in the form of ISCOM and are incorporated herein.
microparticles
In some embodiments of the present invention, there is provided a vaccine or immunogenic composition of the invention comprising microparticles. Microparticles, compositions comprising microparticles and methods for producing microparticles are well known in the art (see Singh et al., [2007 Expert Rev. Vaccines 6 (5): 797-808] and WO 98/033. 487). The term "microparticle" as used herein refers to a particle of about 10 nm to about 10,000 μm in diameter or length, derived from polymeric materials having various molecular weights and, in the case of copolymers such as PLG, various lactide / glycolide ratios. In particular, the microparticles will have a diameter allowing parenteral administration without the needles and capillaries becoming clogged. Microparticles are also known as microspheres. The size of a microparticle is readily determined by techniques well known in the art, such as photon correlation spectroscopy, laser diffractometry and / or scanning electron microscopy. The microparticles usable here will be formed from materials that are sterilizable, non-toxic and biodegradable. Such materials include, but are not limited to, poly (α-hydroxy acid), poly (hydroxybutyric acid), polycaprolactone, polyorthoester, polyanhydride.
liposomes
In some embodiments of the present invention, there is provided a vaccine or immunogenic composition of the invention comprising liposomes. The term "liposomes" refers, in general, to single or multilamellar lipid structures (in particular, 2, 3, 4, 5, 6, 7, 8, 9 or 10 lamellae depending on the number of membranes formed lipid) containing an aqueous inner portion. Liposomes and liposome formulations are well known in the art. Lipids, which are capable of forming liposomes, include all substances having the properties of fats or fat-like properties. The lipids that may constitute lipids in liposomes may be chosen from the group consisting of glycerides, glycerophospholipids, glycerophosphinolipids, glycerophospholipids, sulpholipids, sphingolipids, phospholipids, isoprenolides, steroids, stearins, sterols, archaolipids, synthetic cationic lipids and carbohydrate-containing lipids. The size of the liposomes may vary from 30 nm to several μm depending on the composition of the phospholipids and the method used for the preparation. In particular embodiments of the invention, the size of the liposomes will be in the range of 50 nm to 500 nm, and in other embodiments, from 50 nm to 200 nm. Dynamic scattering of laser light is a method used to measure the size of liposomes which is well known to those skilled in the art. The liposomes suitably contain a neutral lipid, for example a phosphatidylcholine, which is suitably non-crystalline at room temperature, for example egg yolk phosphatidylcholine, dioleoyl phosphatidylcholine (DOPC) or dilauryl phosphatidylcholine. In a particular embodiment, the liposomes of the present invention contain DOPC. The liposomes may also contain a charged lipid which increases the stability of the liposome-saponin structure for liposomes composed of saturated lipids. In this case, the amount of lipid loaded is suitably from 1 to 20% (w / w), preferably from 5 to 10%. The sterol ratio on phospholipid is from 1 to 50% (mole / mole), suitably from 20 to 25% (mole / mole).
saponins
In certain embodiments of the invention, the vaccine or immunogenic composition of the invention comprises a saponin. A saponin particularly suitable for use in the present invention is Quil A and its derivatives. Quil A is a saponin preparation isolated from the Quillaja Saponaria Molina tree of South America, and its adjuvant activity has been described for the first time by Dalsgaard et al. in 1974 ("Saponin adjuvants", Archiv für die gesamte Viruseschung, Vol 44, Springer Verlag, Berlin, p 243-254). HPLC-purified fragments of Quil A that retain adjuvant activity without Quil A-associated toxicity (EP 0 362 278), for example QS7 and QS21 (also known as QA7 and QA21), were isolated by HPLC. QS-21 is a natural saponin derived from the bark of Quillaja saponaria Molina, which induces CD8 + cytotoxic T lymphocytes (CTL), Th1 lymphocytes and a predominant IgG2a antibody response, and is a particular saponin in the context of the present invention. The saponin adjuvant in the immunogenic compositions of the invention is, in particular, an immunologically active fraction of Quil A, such as QS-7 or QS-21, suitably QS-21. In particular embodiments, the vaccines or immunogenic compositions of the invention contain the immunologically active saponin fraction in a substantially pure form. In particular, the vaccines or immunogenic compositions of the invention contain QS-21 in substantially pure form, i.e., QS21 is at least 75% pure, 80%, 85%, 90% pure. for example at least 95% pure, or at least 98% pure.
In a particular embodiment, QS21 is with an exogenous sterol, such as cholesterol for example. Suitable sterols include β-sitosterol, stigmasterol, ergosterol, ergocalciferol and cholesterol. In a further particular embodiment, the adjuvant composition comprises cholesterol as sterol. These sterols are well known in the art, for example cholesterol is described in the Merck Index, 11th Edition, page 341, as a naturally occurring sterol found in animal fats.
In one embodiment, the liposomes of the invention which comprise a saponin conveniently contain a neutral lipid, for example a phosphatidylcholine, which is suitably non-crystalline at room temperature, for example egg yolk phosphatidylcholine, dioleoyl phosphatidylcholine (DOPC) or dilauryl phosphatidylcholine. The liposomes may also contain a charged lipid which enhances the stability of the liposome-QS21 structure for liposomes composed of saturated lipids. In this case, the amount of lipid loaded is suitably 1 to 20% (w / w), especially 5 to 10% (w / w). The sterol ratio on phospholipid is from 1 to 50% (mole / mole), suitably from 20 to 25% (mole / mole).
When the active saponin fraction is QS21, the QS21 / sterol ratio is generally in the range of 1/100 to 1/1 (w / w), suitably 1/10 to 1/1 (w / w). weight), and preferably from 1/5 to 1/1 (w / w). Suitably, an excess of sterol is present, the ratio QS21 / sterol being at least 1/2 (w / w). In one embodiment, the QS21 / sterol ratio is 1/5 (w / w). Suitably, the sterol is cholesterol. Other useful saponins come from the plants Aesculus hippocastanum or Gyophilla struthium. Other saponins that have been described in the literature include escin, which has been described in the Merck index (12th Edition: entry 3737) as a mixture of saponins found in the seeds of horse chestnut, Lat: Aesculus hippocastanum. Its isolation is described by chromatography and purification (Fiedler, Arzneimittel-Forsch., 4, 213 (1953)), and by means of ion exchange resins (Erbring et al., US 3,238,190). Escin fractions were purified and found to be biologically active (Yoshikawa et al., 1996, Chem Pharm Bull (Tokyo), 44 (8): 1454-1464). Sapoalbine from Gypsophila struthium (R. Vochten et al., 1968, J. Pharm.Lab.42: p 213-226) has also been described in connection with the production of ISCOM, for example.
A saponin, such as QS21, can be used in amounts of between 1 and 100 pm per human dose of the adjuvant composition. QS21 can be used at a level of about 50 μg, for example between 40 and 60 μg, suitably between 45 and 55 μg or between 49 and 51 μg, or 50 μg. In a further embodiment, the human dose of the adjuvant composition comprises QS21 at a level of about 25 μg, for example between 20 and 30 μg, suitably between 21 and 29 μg or between 22 and 28 μg. or between 28 and 27 pg or between 24 and 26 pg, or 25 pg.
TLR4 agonist
In some embodiments, the vaccine or immunogenic composition of the invention comprises a TLR4 agonist. By "TLR agonist" is meant a component that is capable of inducing a signaling response via a TLR signaling pathway, either in the form of a direct ligand, or indirectly via generation of an endogenous or exogenous ligand (Sabroe et al., 2003, JIp 1630-5). A TLR4 agonist is capable of inducing a signaling response via a TLR4 signaling pathway. An appropriate example of a TLR4 agonist is a lipopolysaccharide, suitably a nontoxic derivative of lipid A, particularly monophosphorylated lipid A, or more particularly 3-deacylated monophosphoryl lipid A (3D-MPL).
3D-MPL is sold as MPL by GlaxoSmithKline Biologicals and is called MPL or 3D-MPL throughout the document. See, for example, US 4,436,727; U.S. 4,877,611; US 4,866,034 and US 4,912,094. 3D-MPL primarily promotes CD4 + T cell responses with an IFN-γ (Th1) phenotype. 3D-MPL can be produced according to the methods described in GB 2 220 211 A. Chemically, it is a mixture of monophosphorylated lipid A 3-deacylated with 4, 5 or 6 acylated chains. In the composition of the present invention, small particles of 3D-MPL can be used to prepare the aqueous adjuvant composition. 3D-MPL in the form of small particles has a particle size such that it can be sterilized by filtration through a 0.22 μm filter. Such preparations are described in WO 94/21292. Preferably, powdery 3D-MPL is used to prepare the aqueous builder compositions of the present invention. Other TLR4 agonists that can be used are alkyl glucosaminide phosphates (AGPs), such as those disclosed in WO 98/50399 or US Pat. No. 6,303,347 (AGP preparation methods are also disclosed), suitably RC527 or RC529 or pharmaceutically acceptable salts of AGP as disclosed in US 6,764,840. Other suitable TLR-4 agonists are described in WO 03/011 223 and WO 03/099 195, such as compound I, compound II and compound III described on pages 4 and 5 of WO 03/011 223 or on pages 3 and 4 of WO 03/099 195, and in particular the compounds disclosed in WO 03/011 223, such as ER803022, ER803058, ER803732, ER804053, ER804057m ER804058, ER804059, ER804442, ER804680 and ER804764. For example, a suitable TLR-4 agonist is ER804057.
A TLR4 agonist, such as a lipopolysaccharide, such as 3D-MPL, can be used in an amount between 1 and 100 μg per human dose of the adjuvant composition. 3D-MPL can be used at a level of about 50 μg, for example between 40 and 60 μg, suitably between 45 and 55 μg or between 49 and 51 μg or 50 μg. In another embodiment, the human dose of the adjuvant composition comprises 3D-MPL at a level of about 25 μg, for example between 20 and 30 μg, suitably between 21 and 29 μg or between 22 and 28 μg. or between 28 and 27 pg or between 24 and 26 pg, or 25 pg.
Synthetic derivatives of lipid A are known and appear to be TLR 4 agonists, such as, but not limited to: OM174, (2-deoxy-6-o- [2-deoxy-2 - [(R) Dodecanoyloxytetra-decanoylamino] -4-o-phosphono-D-glucopyranosyl] -2- [(R) -3-hydroxytetradecanoylamino] -α-D-glucopyranosyldihydrogenphosphate) (WO 95/14 026) OM 294 DP, ( 3S, 9R) -3 - [(R) -Dodecanoyloxytetradecanoylamino] -4-oxo-5-aza-9 (R) - [(R) -3-hydroxytetradecanoylamino] decan-1,10-diol, 1.10 bis (dihydrogen phosphate) (WO 99/64301 and WO 00/0462) OM 197 MP-Ac DP, (3S, 9R) -3 - [(R) -dodecanoyloxytetradecanoylamino] -4-oxo 5-aza-9 - [(R) -3-hydroxytetrafanoylamino] decan-1,10-diol, 1-dihydrogenphosphate 10- (6-aminohexanoate) (WO 01/46127). Other TLR-4 ligands capable of inducing a signaling response via TLR-4 (Sabroe et al., JI 2003 p 1630-5) are, for example, a lipopolysaccharide derived from a Gram negative and its derivatives, or fragments thereof, particularly a nontoxic derivative of LPS (such as 3D-MPL). Other suitable TLR agonists are: heat shock protein (HSP) 10, 60, 65, 70, 75 or 90; surfactant protein A, hyaluronan oligosaccharides, heparan sulfate fragments, fibronectin fragments, fibrinogen peptides and b-defensin-2, muramyl dipeptide (MDP) or respiratory syncytial virus protein F (RSV). In one embodiment, the TLR agonist is HSP 60, 70 or 90.
TLR agonists
Rather than a TLR4 agonist, other natural or synthetic TLR agonists may be used in the vaccines or immunogenic compositions of the invention. These include, but are not limited to, TLR2, TLR3, TLR5, TLR6, TLR7, TLR8 and TLR9 agonists.
In one embodiment of the present invention, a TLR agonist is used which is capable of inducing a signaling response via TLR-1 (Sabroe et al., JI 2003 p 1630-5). Suitably, the TLR agonist capable of inducing a signaling response via TLR-1 is selected from: triacylated lipopeptides (LP); phenol soluble modulin; an LP of Mycobacterium tuberculosis; S- (2,3-bis (palmitoyloxy) - (2-RS) -propyl) -N-palmitoyl- (R) -Cys- (S) -Ser- (S) -Lys (4) propionic trihydrochloride OH (Pam3Cys) which mimics the acylated N-terminus of a bacterial lipoprotein and Borrelia burgdorfei's LP OspA.
In a further embodiment, a TLR agonist is used which is capable of inducing a signaling response via TLR-2 (Sabroe et al., JI 2003 p 1630-5). Suitably, the TLR agonist capable of inducing a signaling response via TLR-2 is one or more of a lipoprotein, a peptidoglycan, a bacterial lipopeptide derived from M. tuberculosis, B. burgdorferi, T. pallidum, peptidoglycans from species such as Staphylococcus aureus, lipoteichoic acids, mannuronic acids, Neisseria porins, bacterial fimbriae, Yersinia virulence factors, CMV virions, measles haemagglutinin and yeast zymosan.
In a further embodiment, a TLR agonist is used which is capable of inducing a signaling response via TLR-3 (Sabroe et al., JI 2003 p 1630-5). Suitably, the TLR agonist capable of inducing a signaling response via TLR-3 is double-stranded RNA (dsRNA) or polyinosinic-polycytidylic acid (poly IC), a nucleic acid profile. Molecules associated with a viral infection.
In an alternative embodiment, a TLR agonist is used which is capable of inducing a signaling response via TLR-5 (Sabroe et al., JI 2003 p 1630-5). Suitably, the TLR agonist capable of inducing a signaling response via TLR-5 is a bacterial flagellin. The said TLR-5 agonist may be a flagellin or may be a flagellin fragment that retains the agonist activity of TLR-5. Flagellin may comprise a polypeptide selected from the group consisting of H. pylorl; S. typhimurium, V. cholera, S. marcesens, S. flexneri, T. pallidum, L. pneumophilia, B. hurgdorferei; C. difficile, R. meliloti, A. tumefaciens; R. lupine; B. clarridgeiae, P. mirabilis, B. subtilus, L. moncytogenes, P. aeruginoa and E. coli.
In a particular embodiment, the flagellin is selected from the group consisting of S. typhimurium flagellin B (Genbank accession number AF045151), a fragment of S. typhimurium flagellin B, E. coli (access number
Genbank AB028476); a fragment of FliC of E. coli; S. typhimurium FliC flagellin (ATCC14028) and S. typhimurium FliC flagellin fragment.
In a further particular embodiment, said TLR-5 agonist is a truncated flagellin, as described in WO 09/156405, i.e. a flagellin from which the hypervariable domain has been deleted. In one aspect of this embodiment, said TLR-5 agonist is selected from the group consisting of: FliCu174-4oo / FÜCM61-405 and FliCuI 38-405 ·
In a further particular embodiment, said TLR-5 agonist is a flagellin, as described in WO 09/128 950.
In a further embodiment, a TLR agonist is used which is capable of inducing a signaling response via TLR-6 (Sabroe et al., JI 2003 p 1630-5). Suitably, the TLR agonist capable of inducing a signaling response via TLR-6 is a mycobacterial lipoprotein, a diacylated LP and the phenol-soluble modulin. Other TLR6 agonists are described in WO 03/043 572.
In a further embodiment, a TLR agonist is used which is capable of inducing a signaling response via TLR-7 (Sabroe et al., JI 2003 p 1630-5). Suitably, the TLR agonist capable of inducing a signaling response via TLR-7 is a single-stranded RNA (ssRNA), loxoribine, a guanosine analogue at positions N7 and C8, or a imidazoquinoline compound or a derivative thereof. In a particular embodiment, the TLR agonist is imiquimod. Other TLR7 agonists are described in WO 02/085905.
In a further embodiment, a TLR agonist is used which is capable of inducing a signaling response via TLR-8 (Sabroe et al., JI 2003 p 1630-5). Suitably, the TLR agonist capable of inducing a signaling response via TLR-8 is a single-stranded RNA (ssRNA), an imidazoquinoline molecule having antiviral activity, for example the requisimod (R848 ); the requisimod is also able to be recognized by TLR-7. Other TLR-8 agonists that can be used include those described in WO 04/071459.
In a further embodiment, a TLR agonist is used which is capable of inducing a signaling response, such as a TLR agonist comprising a CpG motif. The term "immunostimulatory oligonucleotide" is used herein to refer to an oligonucleotide that is capable of activating a component of the immune system. In one embodiment, the immunostimulatory oligonucleotide comprises one or more unmethylated cytosine-guanosine units (CpG). In a further embodiment, the immunostimulatory oligonucleotide comprises one or more unmethylated thymidine-guanosine units (TG) or may be T-rich. By T-rich is meant that the nucleotide composition of the oligonucleotide comprises more 50, 60, 70 or 80% thymidine. In one embodiment, the oligonucleotide is not an immunostimulatory oligonucleotide and does not include an unmethylated CpG motif. In a further embodiment, the immunostimulatory oligonucleotide is not T-rich and / or does not include an unmethylated TG moiety. The oligonucleotide may be modified to improve stability in vitro and / or in vivo. For example, in one embodiment, the oligonucleotides are modified to include a phosphorothioate backbone, i.e., inter-nucleotide linkages. Other suitable modifications, such as diphosphorothioate, phosphoroamidate and methylphosphonate modifications, as well as oligonucleotide-alternative internucleotide linkages, are well known to those skilled in the art and are encompassed by the invention.
In another embodiment, the vaccines or immunogenic compositions of the invention further comprise an immunostimulant selected from the group consisting of: a TLR-1 agonist, a TLR-2 agonist, a TLR-3 agonist, a TLR-4 agonist, a TLR-5 agonist, a TLR-6 agonist, a TLR-7 agonist, a TLR-8 agonist, a TLR-9 agonist, or a combination thereof.
Calcium composites
In some embodiments, the vaccine or immunogenic composition of the invention comprises a calcium fluoride composite, the composite comprising Ca, F and Z. "Z" represents an organic molecule. The term "composite" refers to a substance that exists in solid form when dry, and that is insoluble or poorly soluble in pure water. In some aspects, Z includes a functional group that forms an anion when ionized. Such functional groups include, but are not limited to, one or more functional groups selected from the group consisting of: hydroxyl, hydroxylate, hydroxo, oxo, N-hydroxylate, hydroaxamate, N-oxide, bicarbonate, carbonate, carboxylate groups fatty acid, thiolate, organic phosphate, dihydrogen phosphate, monohydrogenphosphate, phosphoric acid monoesters, phosphoric acid diesters, phospholipid esters, phosphorothioate, sulphates, hydrogen sulphates, enolate, ascorbate, phosphoascorbate, phenolate and imine olates .
In some aspects, the calcium fluoride composites described herein include Z, where Z is an anionic organic molecule having affinity for calcium and forming a water insoluble composite with calcium and fluoride.
In additional aspects, the calcium fluoride composites described herein include Z, where Z can be categorized as comprising a member of a chemical class selected from the group consisting of: hydroxyl, hydroxylates, a hydroxo, an oxo, an N-hydroxylate, a hydroaxamate, an N-oxide, bicarbonates, carbonates, carboxylates and a dicarboxylate, salts of carboxylic acids, salts of QS21, an extract of Quillaja bark saponaria, an active immunological saponin extract, salts of a saturated or unsaturated fatty acid, oleic acid salts, amino acid salts, thiolates, a thiolactate, a salt of thiol compound, salts cysteine, N-acetyl-cysteine salts, L-2-oxo-4-thiazolidinecarboxylate, phosphates, dihydrogenphosphates, monohydrogenphosphate, phosphoric acid salts, monoesters of phosphoric acids and their salts , the acid diesters p and the salts thereof, esters of 3'-O-deacylated 4'-monophosphoryl lipid A, 3D-MLA esters, MPL, phospholipid esters, DOPC, dioleolyphosphatidic derivatives, phosphates from CpG units, phosphorothioates of the CpG family, sulphates, hydrogen sulphates, sulfuric acid salts, enolates, ascorbates, phosphoascorbate, phenolate, α-tocopherol, imine olates, cytosine, methyl cytosine , uracyl, thymine, barbituric acid, hypoxanthine, inosine, guanine, guanosine, 8-oxo-adenine, xanthine, uric acid, pteroic acid, acid pteroylglutamic, folic acid, riboflavin and lumiflavin. In additional aspects, the calcium fluoride composites described herein include Z, where Z is selected from the group consisting of: N-acetyl cysteine, a thiolactate; an adipate; a carbonate; folic acid; glutathione; and uric acid. In some aspects, the calcium fluoride composites described herein include Z, wherein Z is selected from the group consisting of: N-acetyl cysteine; an adipate; a carbonate; and folic acid. In additional aspects, the calcium fluoride composites described herein include Z, where Z is N-acetyl cysteine, and the composite comprises between 51% Ca, 48% F, and at most 1% N-acetyl cysteine. acetyl cysteine (w / w) and 37% Ca, 26% F, and 37%
N-acetyl cysteine (in weight / weight). In additional aspects, the calcium fluoride composites described herein include Z, where Z is a thiolactate, and the composite comprises between 51% Ca, 48% F, and at most 1% thiolactate (by weight). weight) and 42% Ca, 30% F, and 28%
thiolactate (in weight / weight). In additional aspects, the calcium fluoride composites described herein include Z, where Z is adipate, and the composite comprises between 51% Ca, 48% F, and at most 1% adipate (by weight). / weight) and 38% Ca, 27% F, and 35% adipate (w / w). In additional aspects, the calcium fluoride composites described herein include Z, where Z is a carbonate, and the composite comprises between 51% Ca, 48% F, and at most 1% carbonate (by weight). weight) and 48% Ca, 34% F, and 18% carbonate (w / w). In additional aspects, the calcium fluoride composites described herein include Z, where Z is folic acid, and the composite comprises between 51% Ca, 48% F, and at most 1% folic acid. (w / w) and 22% Ca, 16% F, and 62% folic acid (w / w). In additional aspects, the calcium fluoride composites described herein include Z, where Z is glutathione, and the composite comprises between 51% Ca, 48% F, and at most 1% glutathione (w / w). weight) and 28% Ca, 20% F, and 52% glutathione (w / w). In additional aspects, the calcium fluoride composites described herein include Z, where Z is uric acid, and the composite comprises between 51% Ca, 48% F, and at most 1% acid. uric (weight / weight) and 36% Ca, 26% F, and 38% uric acid (w / w).
Aluminum salts
In one embodiment, the vaccine or immunogenic composition of the invention comprises an aluminum salt. Suitable aluminum salt adjuvants are well known to those skilled in the art and include, but are not limited to, aluminum phosphate, aluminum hydroxide, or a combination thereof. Suitable aluminum salt adjuvants include, but are not limited to, Rehydragel ™ HS, Alhydrogel ™ 85, Rehydragel ™ PM, Rehydragel ™ AB, Rehydragel ™ HPA, Rehydragel ™ LV, and the like. Alhydrogel ™ or a combination thereof. In particular, the methods of the invention are used to determine the endotoxin content of Adjuphos, Rehydragel ™ HS (3% aluminum hydroxide in water [General Chemical]) or Alhydrogel ™ 85 (Brenntag BioSector [Denmark]).
In particular, the aluminum salts can have a protein adsorption capacity of between 2.5 and 3.5, 2.6 and 3.4, 2.7 and 3.3 or 2.9 and 3.2, 2 , 5 and 3.7, 2.6 and 3.6, 2.7 and 3.5, or 2.8 and 3.4 protein (BSA) / ml of aluminum salt. In a particular embodiment of the invention, the aluminum salt has a protein adsorption capacity of between 2.9 and 3.2 mg of BSA / mg of aluminum salt. The adsorption capacity of the aluminum salt proteins can be measured by any means known to those skilled in the art. The adsorption capacity of the aluminum salt proteins can be measured using the method described in Example 1 of WO 12/136 823 (which uses BSA) or variations thereof.
The aluminum salts described herein (i.e., having the adsorptive capacity of the proteins described herein) may have a crystal size between 2.8 and 5.7 nm, such as as measured by X-ray diffraction, for example 2.9 to 5.6 nm, 2.8 to 3.5 nm, 2.9 to 3.4 nm or 3.4 to 5.6 nm or 3.3 at 5.7 nm, as measured by X-ray diffraction. X-ray diffraction is well known to those skilled in the art. In a particular embodiment of the invention, the size of the crystals is measured using the method described in Example 1 of WO 12/136 823 or variations thereof.
Modes of administration
The pro-resolution mediator can be administered at the same time, before or after administration of the vaccine or immunogenic composition. Therefore, the invention provides pro-resolution mediators of the invention for use in reducing the reactogenicity induced by administration of a vaccine or immunogenic composition, wherein the pro-resolution mediator (s) are administered. 5, 10, 20, 30, 45 minutes or more, or 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 hours or more before administration of the vaccine or immunogenic composition, particularly between 30 minutes and 3 hours, especially about 1 hour before administration of the vaccine or immunogenic composition. The invention further provides, in certain embodiments, pro-resolution mediators of the invention for use in reducing the reactogenicity induced by the administration of a vaccine or immunogenic composition, the pro-resolution mediators being administered concomitantly with said vaccine or said immunogenic composition. The pro-resolution mediators can be administered at the same time by the same route of administration or by a different route. By "concomitantly" is meant more or less than 5 minutes maximum of the time of administration of the vaccine or immunogenic composition, for example up to 1, 2, 3 or 4 minutes before or after administration of the vaccine or immunogenic composition. In embodiments in which the pro-resolution mediator (s) of the invention are administered concomitantly with the vaccine or immunogenic composition by the same route of administration, said proresolution mediator (s) may be formulated appropriately with the antigen component and / or the adjuvant component of the vaccine or immunogenic composition. Therefore, in particular embodiments, the pro-resolution mediator (s) of the invention are formulated with the antigenic component of the vaccines or immunogenic compositions of the invention. In additional particular embodiments, the pro-resolution mediator (s) of the invention are formulated with the adjuvant component of the vaccines or immunogenic compositions of the invention. The invention further provides, in certain embodiments, pro-resolution mediators of the invention for use in reducing the reactogenicity induced by the administration of a vaccine or immunogenic composition, the pro-resolution mediators being administered 5, 10, 20, 30, 45 minutes or more, or 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 hours or more after administration of the vaccine or the immunogenic composition, particularly between 30 minutes and 3 hours, particularly at about 1 hour.
The pro-resolution mediator described herein may be administered by any route of administration. The route of administration may be identical to or different from that of the vaccine / immunogenic composition. Therefore, the invention provides pro-resolution mediators of the invention for use in reducing the reactogenicity induced by administration of a vaccine or immunogenic composition, the proresolution mediator being administered by the same route. as the vaccine or the immunogenic composition. The invention also provides proresolution mediators of the invention for use in reducing the reactogenicity induced by the administration of a vaccine or immunogenic composition, the pro-resolution mediator described herein being administered by a different route from that of the vaccine or the immunogenic composition.
A pro-resolution mediator described herein can be administered orally, sublingually, intramuscularly, intradermally (e.g., a transdermal patch with micro-projections) or transdermally (e.g., ointment or cream). The invention further provides proresolution mediators of the invention for use in reducing the reactogenicity induced by the administration of a vaccine or immunogenic composition, the pro-resolution mediator described herein may be administered at the same site on the patient (e.g., the upper arm), but by different routes of administration (particularly, wherein the vaccine or immunogenic composition is delivered intramuscularly or intradermally, and wherein the proresolution mediator is delivered transdermally (e.g., an ointment or cream) .The cream or ointment comprising the pro-resolution mediator may be administered prior to, concurrently with, or after administration of the vaccine / regimen. immunogenic composition by intradermal or intramuscular administration.
Pharmaceutically acceptable compositions
The pro-resolution mediator (s), vaccine and immunogenic compositions of the invention are pharmaceutically acceptable. They may comprise components in addition to the proresolution mediator (s), the antigen (s) and / or adjuvant, for example they generally comprise one or more pharmaceutical carriers and / or excipients.
The compositions may include preservatives, such as thiomersal or 2-phenoxyethanol. In particular embodiments, the vaccine or immunogenic compositions of the invention are substantially free (i.e., less than 5 μg / ml) of mercury-based material, e.g., free of thiomersal. In particular, the compositions are free of mercury and any preservative.
The compositions of the invention may be isotonic and may therefore comprise a physiological salt, such as a sodium salt. Sodium chloride (NaCl) is preferred, which may be present between 1 and 20 mg / ml. Other salts that may be present include potassium chloride, potassium dihydrogenphosphate, dehydrated disodium phosphate, magnesium chloride, calcium chloride.
The compositions generally have an osmolality of between 200 mOsm / kg and 400 mOsm / kg, preferably between 240 and 360 mOsm / kg, and in particular between 290 and 310 mOsm / kg.
The compositions of the invention may comprise one or more buffers. Conventional buffers include: a phosphate buffer; a Tris buffer; a borate buffer; a succinate buffer; a histidine buffer; or a citrate buffer. The buffers will generally be included in the range of 5 to 20 mM.
The pH of the compositions of the invention may be between 5.0 and 8.1, and more generally between 6.0 and 8.0, for example between 6.5 and 7.5, or between 7.0 and 7.0. , 8.
Kit of the invention
The pro-resolution mediator (s), antigen (s) and / or adjuvant (s) may be prepared extemporaneously at the time of administration. Therefore, the invention provides kits comprising the pro-resolution mediator (s), the antigen (s) and / or adjuvant ready for mixing. The kits allow the pro-resolution mediator (s), antigen (s) and / or adjuvant to be maintained separately until the moment of use. This is particularly important if the proresolution mediator is to be administered at a different time or by a different route of administration, for example.
Therefore, the present invention provides kits comprising i) an antigen as described herein; and ii) a pro-resolution mediator as described in this document. The present invention also provides kits comprising i) an adjuvant as described herein; and ii) a pro-resolution mediator as described in this document. The present invention further provides kits comprising i) an antigen as described herein; ii) an adjuvant as described herein; and iii) a pro-resolution mediator as described in this document.
The components are physically separated from each other within a kit, and this separation can be achieved in a variety of ways. For example, the two components may be in two separate containers, for example vials. The contents of the two vials can then be mixed, for example by removing the contents of one vial and adding it to the other vial, or by separately removing the contents of the two vials and mixing them in a third container (for example). example, a bottle).
In one particular embodiment, one of the kit components is in one syringe and the other is in a container, such as a vial. The syringe can be used (for example, with a needle) to insert its contents into the second container for mixing purposes, and the mixture can then be drawn into the syringe. The mixed contents of the syringe can then be administered to a patient, traditionally via a new sterile needle. Packaging a component in a syringe eliminates the need to use a separate syringe for administration to the patient. In another preferred arrangement, the two kit components are held together but separately in the same syringe, for example a dual chamber syringe. When the syringe is actuated (for example, when administering to a patient), then the contents of both chambers are mixed. This arrangement avoids the need for a separate mixing step at the time of use.
The components of the kit may be in aqueous form. In some embodiments, a component, such as the pro-resolution antigen (s) or mediator (s), is in a dry form (for example, in freeze-dried form), the other component being in aqueous form. The two components can be mixed to reactivate the dry component and provide an aqueous composition that can be administered to a patient. A lyophilized component will generally be located in a vial rather than a syringe.
The dry components may include stabilizers, such as lactose, sucrose or mannitol, and mixtures thereof, for example, lactose / sucrose mixtures, sucrose / mannitol mixtures, and the like. One possible arrangement uses an aqueous adjuvant component in a pre-filled syringe and a lyophilized antigen component in a vial.
Examples
Example 1 Effect of resolvin El (RvE1) on the profile of local immune cells induced by the injection of adjuvants
Mice (n = 6 / g) were injected intramuscularly with PBS, adjuvant AS03 (oil-in-water emulsion) or AS01B adjuvant (MPL + QS21 in liposomes). 1 hour and 6 hours later, synthetic RvE1 (0.2 μg / dose) or PBS was also administered intramuscularly. Muscles were collected 24 hours after adjuvant / PBS administration and local immune cells were extracted. The cells were stained with the following antibodies: Ly6c, SiglecF, CD90.2, Ly6G, CD11b, CD11c, HLA DR, CD45 and CD19, and analyzed by flow cytometry. The recruitment of the cells is expressed in number of cells per muscle. The plotted area represents the average obtained for a specific subpopulation. A two-way ANOVA analysis was performed to compare the different groups. *: p <0.05. The results are shown in Figure 1. The chemical structure of the synthetic RvE1 used in the present experiment is identical to the chemical structure of RvE1 disclosed in Buckley et al. (Proresolving Lipid Mediators and Mechanisms in the Resolution of Acute Inflammation, 2014, Immunity 40: 315-327). Results - Conclusions The injection of each adjuvant, AS03 and AS01B, resulted in the recruitment of immune cells at the injection site, compared with PBS alone. Injection of RvE1 after the AS03 adjuvant injection resulted in significantly reduced recruitment of all types of immune cells tested. The same conclusion is reached with the injection of RvE1 after injection of ASOlB, indicating that the RvE1 used in this experiment was biologically active, being able to modulate the immune cell profile at the level of injection site. It is also interesting to note that the injection of each adjuvant alone gave a different profile of immune cells at the injection site. In addition, the RvE1 itself, with PBS alone, appeared to have only a minimal effect on the immune cell profile at the injection site, compared to PBS alone.
Example 2 Effect of 7-Marinesin-1 (MaR1) on the Profile of Local Immune Cells Induced by the Injection of Adjuvants
Mice (n = 6 / g) received by intramuscular injection a synthetic 7-maresin-1 (MaR1) (5 ng / dose) or PBS as a control. 1 hour later, the mice received AS01B adjuvant or PBS intramuscularly. The muscles were collected 4 hours and 24 hours after the administration of 7-marinesin-1 / PBS and the local immune cells were extracted. The cells were stained with the following antibodies: Ly6c, SiglecF, CD90.2, Ly6G, CD11b, CD11c, HLA DR, CD45 and CD19, and analyzed by flow cytometry. The recruitment of the cells is expressed in number of cells per muscle. The plotted area represents the average obtained for a specific subpopulation. A two-way ANOVA analysis was performed to compare the different groups. *: p <0.05. The results are shown in Figure 2. The chemical structure of the synthetic MaR1 used in the present experiment is identical to the chemical structure of the MaRl disclosed in Buckley et al. (Proresolving Lipid Mediators and Mechanisms in the Resolution of Acute Inflammation, 2014, Immunity 40: 315-327). Results - Conclusions
When MaR1 is injected before the AS01B adjuvant injection, the most significant modulation effect observed on the immune cell profile at the injection site was obtained 24 hours after the injection, which confirms that in this experiment, MaRl was biologically active. The main changes observed were an increase in B-cell and T-cell recruitment at the site, which is a feature of a late resolution phase. This suggests that observation of a modulation effect of MaR1 may depend on the time after injection, and that 4 hours after injection may be too early to detect an effect.
Example 3 Effect of resolvin El (RvE1) on the profile of cytokines induced by the injection of adjuvants
Mice (n = 6 / g) were injected intramuscularly with PBS, AS03 adjuvant or AS01B adjuvant. 1 hour and 6 hours later, the same synthesis RvE1 as that used in Example 1 (0.2 μg / dose), or PBS was also administered intramuscularly. The muscles were collected 4 hours (A) and 24 hours (B) after administration of the adjuvant or PBS and frozen at -70 ° C. The muscles were thawed and homogenized, and the levels of cytokines (TNFα, IL-6, IL1b and IFNγ) and chemokines (G-CSF, MCP-1, KC and ΜΙΡ-la) in the clarified homogenates were measured by CBA (Becton Dickinson). The cytokine / chemokine concentrations are expressed in μg / ml. The plotted area represents the average obtained for a specific cytokine / chemokine. A two-way ANOVA analysis was performed to compare the different groups. *: p <0.05 (see Figure 3). Results - Conclusions The injection of each of the adjuvants, AS03 and AS01B, resulted in an increase in the concentration of cytokines / chemokines at the injection site, relative to PBS alone, to different degrees. With regard to adjuvant AS01B, no significant modulation effect was observed when the RvE1 was injected after adjuvant injection at the two time points tested, that is, 4 hours and 24 hours. hours after the adjuvant injection (see Box A and Box B, respectively, in Figure 3). With respect to the adjuvant AS03, a significant modulation effect was observed when the RvE1 was injected after the adjuvant injection 4 hours after the adjuvant injection, whereas no significant effect occurred. was observed 24 hours after the injection of the adjuvant. These results indicate that the RvE1 used in this experiment was biologically active, being able to modulate the concentration of cytokines / chemokines at the injection site. These results also suggest that the ability of RvE1 to modulate the profile of local cytokines / chemokines may differ depending on the type of adjuvant that is injected and / or that the modulation effect may be observed at different time points after the injection. adjuvant.
Example 4 Effect of 7-Marinesin-1 (MaR1) on the Profile of Local Cytokines Induced by the Injection of Adjuvants
Mice (n = 6 / g) received by intramuscular injection the same synthetic MaR1 as used in Example 2 (5 ng / dose), or PBS as a control. 1 hour later, the mice received AS01B adjuvant or PBS intramuscularly. Muscles were collected 4 hours and 24 hours after administration of MaRl / PBS and frozen at -70 ° C. The muscles were thawed and homogenized, and the levels of cytokines (TNFα, IL-6, IL1b and IFNγ) and chemokines (G-CSF, MCP-1, KC and ΜΙΡ-la) in the clarified homogenates were measured by CBA (Becton Dickinson). The cytokine / chemokine concentrations are expressed in μg / ml. The plotted area represents the average obtained for a specific cytokine / chemokine. A two-way ANOVA analysis was performed to compare the different groups. *: p <0.05 (see Figure 4). Results - Conclusions The injection of each AS01B adjuvant resulted in an increase in cytokine / chemokine concentration at the injection site, relative to PBS alone. A significant modulation transient effect was observed when MaR1 was injected prior to adjuvant injection 4 hours after MaR1 injection. These results indicate that the MaR1 used in this experiment was biologically active, being able to modulate the profile of local cytokines / chemokines at the injection site.
Example 5 - No Effect of 7-Marinesin-1 (MaR1) on Specific T-cell Responses Induced by Vaccine Injection
Mice (n = 30 / g) received by intramuscular injection the same synthetic MaR1 as used in Example 2 (5 ng / dose), or PBS as a control. One hour later, the mice were vaccinated with OVA (ovalbumin) and HBS (hepatitis B surface antigen) antigens resuspended in AS01B adjuvant or in PBS. This vaccination schedule was repeated 15 days later.
The spleens were removed 7 days after the second immunization, and the immune cells were extracted and stimulated overnight with the OVA (A) and HBS (B) peptides. After surface staining with the anti-CD4 and anti-CD8 antibodies, the cells were labeled intracellularly with an anti-IL-2, an anti-IFN-gamma and an anti-TNF-α and analyzed by flow cytometry. . The results are expressed as a percentage of CD4 + or CD8 + T lymphocytes expressing at least 2 cytokines among those tested. Each point represents an individual value, the bar represents the + / - ET mean (see Figure 5). Results - Conclusions The injection of both the OVA and HBS antigens in combination with the AS01B adjuvant induced a significant CD4 + T lymphocyte response, as well as a significant CD8 + T lymphocyte response, compared with the injection. antigens alone. When MaR1 was injected prior to the injection of the antigens in combination with the AS01B adjuvant, nor the CD4 + T lymphocyte response, neither the CD8 + T lymphocyte response was observed to be statistically inferior to the responses induced by antigens and adjuvant alone (see Boxes A and B in Figure 5). These results indicate that the injection of MaR1 did not have a negative impact on the T cell immune response induced by the injection of an adjuvanted vaccine.
Example 6 - No Effect of 7-Marinesin-1 (MaR1) on Specific Antibody Responses Induced by Vaccine Injection
Mice (n = 30 / g) received by intramuscular injection the same synthetic MaR1 as used in Example 2 (5 ng / dose), or PBS as a control. 1 hour later, the mice were vaccinated with OVA and HBS antigens resuspended in AS01B adjuvant or in PBS. This vaccination schedule was repeated 15 days later. The sera were taken 7 days after the second immunization. Anti-OVA (A) or anti-HBs (B) antibodies were detected by ELISA. The antibody titers are expressed in ng / ml. Each point represents an individual value; the bar represents the + / - ET mean (see Figure 6). Results - Conclusions The injection of both OVA and HBS antigens in combination with AS01B adjuvant induced a significant anti-OVA antibody response (see Box A in Figure 6) and a significant anti-OVA response. -HBS (see frame B of Figure 6), compared to the injection of the antigens alone. When MaR1 was injected before the injection of the antigens in combination with AS01B adjuvant, or the anti-OVA antibody response (see frame A of FIG. 6), neither the anti-HBS antibody response was observed to be statistically inferior to the responses induced by antigens and adjuvant alone. These results indicate that the injection of MaR1 did not have a negative impact on the antibody-type immune response induced by the injection of an adjuvanted vaccine.
General Conclusions • Intramuscular administration of a small concentration of pro-resolution mediators, such as RvE1 and MaR1, may modulate the immune cell recruitment profile, as well as the local cytokine profile (Figures 1, 2, 3 and 4). . • The timing of administration and the nature of the pro-resolution mediator administered may modulate the local inflammatory pattern induced by the adjuvants differently (Figures 1, 2, 3 and 4). The primary local cytokines modulated by the administration of pro-resolution mediators are IL-6 and MCP-1 (Figure 3A, Figure 4). • The main local immune cells modulated by the administration of proresolution mediators are lymphoid cells and monocytes (Figures 1 and 2). • The pro-resolution mediators can modulate the local inflammatory profile induced by two different adjuvants (AS01B and AS03), said modulation and its extent may differ depending on the adjuvant and if we take into account the recruitment profile of local immune cells or the profile of local cytokines. • Local administration of proresolution mediators has no impact on specific T-cell and antibody-to-vaccine responses (Figures 5 and 6).
Of course, the invention is not limited to the embodiments described above and shown, from which we can provide other modes and other embodiments, without departing from the scope of the invention. .

权利要求:
Claims (50)
[1]
A pro-resolution mediator for use in reducing the reactogenicity induced by administration of a vaccine or immunogenic composition comprising at least one antigen.
[2]
The pro-resolution mediator of claim 1, wherein said pro-resolution mediator is a lipid.
[3]
The pro-resolution mediator of claim 1 or claim 2, wherein said pro-resolution mediator is of natural origin.
[4]
The pro-resolution mediator of claim 1 or claim 2, wherein said pro-resolution mediator is synthetic.
[5]
A pro-resolution mediator according to any preceding claim, wherein the proresolution mediator is derived from polyunsaturated fatty acids (PUFAs).
[6]
A pro-resolution mediator according to any preceding claim, wherein the proresolution mediator is derived from PUFA ω-3 eicosapentaenoic acid (EPA), PUFA ω-3 docosahexaenoic acid (DHA) or PUFA ω-6 arachidonic acid (AA ).
[7]
A pro-resolution mediator according to any preceding claim, wherein the proresolution mediator is selected from the group consisting of: a resolvin (E-series or D-series), a marine, a lipoxin, a protectin, or any combination two or more of them.
[8]
A pro-resolution mediator according to any preceding claim, wherein the proresolution mediator is selected from the group consisting of: RvE1, RvE2, RvE3, RvD1, RvD2, RvD3, RvD4, MaR1, PD1 / NPD1, 17-HDHA and LXA4 , or a functional analogue thereof, or any combination of two or more thereof.
[9]
A pro-resolution mediator according to any preceding claim, wherein the proresolution mediator is MaR1, or a functional analogue thereof.
[10]
The pro-resolution mediator according to any one of claims 1 to 8, wherein the pro-resolution mediator is RvE1, or a functional analogue thereof.
[11]
A pro-resolution mediator according to any preceding claim, wherein the proresolution mediator is administered 5, 10, 20, 30, 45 minutes or more, or 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 hours or more before administration of the vaccine or immunogenic composition, particularly between 30 minutes and 3 hours, especially about 1 hour.
[12]
A pro-resolution mediator as claimed in any one of claims 1 to 10, wherein the pro-resolution mediator is administered concomitantly with said vaccine or said immunogenic composition.
[13]
The pro-resolution mediator according to any one of claims 1 to 10, wherein the pro-resolution mediator is administered 5, 10, 20, 30, 45 minutes or more, or 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 hours or more after administration of the vaccine or immunogenic composition, particularly between 30 minutes and 3 hours, especially about 1 hour.
[14]
A pro-resolution mediator according to any preceding claim, wherein the proresolution mediator is administered by the same route as the vaccine or immunogenic composition.
[15]
The pro-resolution mediator of any one of claims 1 to 13, wherein the pro-resolution mediator is administered by a route different from that of the vaccine or immunogenic composition.
[16]
A pro-resolution mediator according to any preceding claim, wherein the proresolution mediator is administered orally, sublingually, intramuscularly, intradermally (e.g., a transdermal patch with micro-projections) or transdermally (e.g., an ointment or cream).
[17]
The pro-resolution mediator of claim 15 or 16, wherein the proresolution mediator is administered at the same site but by different routes of administration.
[18]
The pro-resolution mediator of claim 17, wherein the vaccine or immunogenic composition is delivered intramuscularly or intradermally, and wherein the pro-resolution mediator is delivered transdermally.
[19]
The pro-resolution mediator of any preceding claim, wherein the vaccine or immunogenic composition comprises an adjuvant.
[20]
The pro-resolution mediator of claim 19, wherein the adjuvant is selected from the group consisting of: an oil-in-water emulsion (e.g., MF59 or AS03), liposomes, a saponin, a TLR2 agonist , a TLR3 agonist, a TLR4 agonist, a TLR5 agonist, a TLR6 agonist, a TLR7 agonist, a TLR8 agonist, a TLR9 agonist, aluminum salts, microparticles, ISCOMs, composites calcium fluoride or any combination of two or more thereof.
[21]
The pro-resolution mediator of claim 19 or 20, wherein the adjuvant comprises a saponin and / or a TLR4 agonist.
[22]
The pro-resolution mediator of claim 20 or 21, wherein the saponin is a Quil A derivative.
[23]
The pro-resolution mediator of claim 22, wherein the Quil A derivative is QS21.
[24]
The pro-resolution mediator of claim 20 or 21, wherein the TLR4 agonist is a detoxified lipopolysaccharide.
[25]
The pro-resolution mediator of claim 24, wherein the detoxified lipopolysaccharide is 3D-MPL.
[26]
The pro-resolution mediator of any one of claims 20 to 25, wherein the saponin and / or TLR4 agonist is in a liposomal formulation.
[27]
The pro-resolution mediator of any one of claims 21 to 26, wherein the adjuvant further comprises a TLR9 agonist, particularly a CpG oligonucleotide.
[28]
The pro-resolution mediator of claim 19 or 20, wherein the adjuvant is an oil-in-water emulsion.
[29]
The pro-resolution mediator of claim 28, wherein the oil-in-water emulsion comprises squalene and a polyoxyethylene sorbitan monooleate (POLYSORBATE 80).
[30]
The pro-resolution mediator of claim 29, wherein the oil-in-water emulsion further comprises a sorbitan trioleate (SPAN 85) or alpha-tocopherol.
[31]
31. The pro-resolution mediator of claim 19 or 20, wherein the adjuvant is an aluminum salt, particularly aluminum hydroxide or aluminum phosphate.
[32]
The pro-resolution mediator of claim 31, wherein the adjuvant further comprises a TLR4 agonist.
[33]
33. The pro-resolution mediator according to claim 32, wherein the TLR4 agonist is adsorbed on the aluminum salt, in particular more than 80, 85, 90, 95% of the TLR4 agonist is adsorbed on the salt. aluminum.
[34]
The pro-resolution mediator of claim 32 or 33, wherein the TLR4 agonist is a detoxified lipopolysaccharide.
[35]
The pro-resolution mediator of claim 34, wherein the detoxified lipopolysaccharide is 3D-MPL.
[36]
A pro-resolution mediator according to any preceding claim, wherein the antigen is selected from the group consisting of: a whole organism, a polypeptide, a polysaccharide, a peptide, a nucleic acid and a protein-polysaccharide conjugate, or n any combination of two or more of these.
[37]
A pro-resolution mediator according to any preceding claim, wherein the antigen is derived from an organism selected from the group consisting of: viruses, bacteria, parasites and fungi, or any combination of two or more than two of these.
[38]
38. A pro-resolution mediator according to any preceding claim, wherein the antigen is derived from a virus.
[39]
39. The pro-resolution mediator according to claim 38, wherein the virus is selected from the group consisting of: influenza virus, RSV, HPV, measles virus, rubella virus, influenza virus. mumps, HCMV, VZV, dengue virus, poliovirus, HIV, HBV, Ebola and rotavirus, or any combination of two or more of these.
[40]
40. A pro-resolution mediator according to any one of claims 1 to 37, wherein the antigen is derived from a bacterium.
[41]
41. The pro-resolution mediator according to claim 40, wherein the bacterium is selected from the group consisting of: B. pertussis, S. Pneumoniae and N. Meningitidis, or any combination of two or more of these this.
[42]
42. A pro-resolution mediator according to claim 41, wherein the antigen is the whole bacterial B. pertussis antigen (Pw antigen), optionally in combination with tetanus toxoid (T) and / or diphtheria toxoid ( D).
[43]
43. The pro-resolution mediator according to claim 41, wherein the antigen is a capsular saccharide of S. pneumoniae derived from a serotype selected from the group consisting of: serotypes 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, or any combination of two or more of those optionally, conjugated to a carrier protein.
[44]
44. The pro-resolution mediator according to claim 41, wherein the antigen is a capsular saccharide of N. Meningitidis derived from a serogroup selected from the group consisting of: serogroup A (MenA), serogroup C (MenC), serogroup Y (MenY), and serogroup W-135 (MenW), or any combination of two or more thereof, optionally conjugated to a carrier protein.
[45]
45. The pro-resolution mediator according to claim 41, wherein the antigen is derived from N. Meningitidis serogroup B and is a polypeptide selected from the group consisting of: NadA protein, NHBA protein, fHBP protein, GNA1030 protein , and the GNA2091 protein, or any combination of two or more thereof, optionally in combination with an outer membrane vesicle (OMV) derived from N. Meningitidis serogroup B.
[46]
46. An immunogenic vaccine or composition comprising an antigen and a pro-resolution mediator as defined in any one of claims 1 to 10, the antigen is as defined in any one of claims 36 to 45 and an adjuvant such as defined in any one of claims 20 to 35.
[47]
47. Kit comprising i) an antigen as defined in any one of claims 36 to 45; and ii) a pro-resolution mediator.
[48]
48. Kit comprising i) an adjuvant as defined in any one of claims 20 to 35; and ii) a pro-resolution mediator as defined in any one of claims 1 to 10.
[49]
49. Kit comprising i) an antigen as defined in any one of claims 36 to 45; (ii) an adjuvant as defined in any one of claims 20 to 35; and iii) a proresolution mediator as defined in any one of claims 1 to 10.
[50]
50. Use of a pro-resolution mediator according to any one of claims 1 to 10 in the preparation of a medicament for reducing the reactogenicity induced by the administration of a vaccine or immunogenic composition as defined in claim 46.

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同族专利:

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公开号 | 公开日

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GB201522132D0|2016-01-27|

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EP3389708A1|2018-10-24|

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BE1024094A1|2017-11-14|

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US11045542B2|2021-06-29|

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WO2017102703A1|2017-06-22|

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US20180360956A1|2018-12-20|

引用文献:

---

公开号 | 申请日 | 公开日 | 申请人 | 专利标题

---

WO1996033739A1|1995-04-25|1996-10-31|Smithkline Beecham Biologicals S.A.|Vaccines containing a saponin and a sterol|

---

WO2017210604A1|2016-06-03|2017-12-07|Thetis Pharmaceuticals Llc|Compositions and methods relating to salts of specialized pro-resolving mediators of inflammation|

---

WO2020023872A1|2018-07-27|2020-01-30|Children's Medical Center Corporation|Use of toll-like receptor 2agonist for modulating human immune response|

---

KR20200046458A|2018-10-24|2020-05-07|아모레퍼시픽|Composition for skin barrier|

法律状态:
2018-02-08| FG| Patent granted|Effective date: 20171116 |

优先权:

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申请号 | 申请日 | 专利标题

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GBGB1522132.8A|GB201522132D0|2015-12-15|2015-12-15|Vaccine|

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GB1522132.8|2015-12-15|

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