METHODS AND COMPOSITIONS FOR INDUCING PROTECTIVE IMMUNITY AGAINST FILOVIRUS DISEASE AND / OR INFECTI

专利摘要:
Compositions, vaccines and methods for inducing protective immunity against filovirus disease and / or infection, particularly protective immunity against Ebolavirus infection are described. The vaccine contains a first composition comprising an immunologically effective amount of at least one human adenoviral vector comprising a first nucleic acid encoding a first antigenic protein of a filovirus, and a pharmaceutically acceptable carrier; and a second composition comprising an immunologically effective amount of a simian adenoviral vector comprising a second nucleic acid encoding a second filovirus antigenic protein, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition.
公开号:BE1023877B1
申请号:E2016/5376
申请日:2016-05-23
公开日:2017-09-01
发明作者:Hoof Johann Jules Urbain Van；Moncef Slaoui；Ripley W. Ballou
申请人:Glaxosmithkline Biologicals Sa；Janssen Vaccines & Prevention B.V.；Crucell Holland B.V.；
IPC主号:

专利说明:

TITLE OF THE INVENTION
METHODS AND COMPOSITIONS FOR INDUCING PROTECTIVE IMMUNITY AGAINST FILOVIRUS DISEASE AND / OR INFECTION
REFERENCE TO RELATED APPLICATIONS (001) This application claims the priority of U.S. Provisional Patent Application No. 62/164909 filed May 21, 2015, the disclosure of which is hereby incorporated by reference in its entirety.
REFERENCE TO THE LISTING OF SEQUENCES SUBMITTED BY WAY
ELECTRONIC (002) This application contains a sequence listing which is submitted electronically via EPS-Web in the form of an ASCII sequence listing with the file name "688097-70WO Sequence Listing", creation date 20 May 2016 and having a size of about 15 kb. The listing of sequences submitted via EFS-Web is part of the description and is hereby incorporated in its entirety by reference.
FIELD OF THE INVENTION (003) The present invention relates to compositions, vaccines and methods for inducing protective immunity against filovirus disease and / or infection, particularly protective immunity against Ebolavirus infection. .
BACKGROUND OF THE INVENTION (004) Marburg viruses (MARV) and Ebola viruses (EBOV), such as Ebolavirus Zaire, are linked to outbreaks of Marburg haemorrhagic fever (MHF) and Ebola virus diseases. (EVD) particularly lethal in humans and primates in Africa, North Africa and Europe. These viruses are filoviruses that are known to infect humans and non-human primates, with serious health consequences, including fatal outcomes. Filovirus infections have resulted in mortality rates of up to 90% in humans. EBOV infections cause EVD with a fatal outcome often occurring within 7 to 10 days after infection. EVD occurs as an acute febrile syndrome manifested by sudden fever, nausea, vomiting, diarrhea, maculopapular rash, malaise, prostration, generalized signs of increased vascular permeability, coagulation abnormalities, and a disturbance of the innate immune response. Much of the disease appears to be caused by a disruption of innate immune responses to infection and by replication of the virus in vascular endothelial cells, resulting in death of host cells and destruction of the endothelial barrier. Filovirus can be spread by aerosol of small particles or by direct contact with infected blood, infected organs and infected body fluids, of human origin or from non-human primates. It is reported that infection with a single virion is sufficient to cause EVD in humans. There is currently no approved therapeutic agent or vaccine for the treatment and prevention of EVD, although two candidate vaccines have arrived to a Phase 2/3 clinical trial underway in Liberia. Supportive treatment remains the primary medical intervention for individuals who have been infected with FIVVs and / or have EVD. (005) Individuals remain infectious for Ebola virus disease (EVD) as long as their blood and secretions contain the virus. The high infectiousness of blood and secretions exposes health personnel to a particularly high risk during outbreaks, and direct contact with the bodies of deceased victims may also play a role in the transmission of the virus. The incubation period of EVD is 2 to 21 days (7 days on average depending on the strain), followed by a severe acute viral illness, often characterized by the rapid onset of nonspecific symptoms, such as fever, extreme fatigue, pharyngitis, gastrointestinal disorders, abdominal pain, anorexia, headache, myalgia and / or arthralgia. (006) A primary antibody response (IgM) can be detected in the blood of infected persons 2 to 9 days after infection, while IgG antibodies appear approximately 17 to 25 days after infection. This IgG response coincides with the recovery phase. In EVD survivors, both humoral and cellular immunity are detected, but their relative contribution to protection is unknown (Sullivan et al., Ebola virus pathogenesis: implications for vaccines and therapies, J. Virol 2003 ; 77: 9733-7). (007) As the cause of severe human illness, filoviras continue to be a source of concern for both natural infections and possible agents of bioterrorism. The reservoir of filoviras in nature has not yet been definitively identified. Four subtypes of Ebolaviras have been described as causing EVD, namely those in episodes in Zaire, Sudan, the Bundibugyo region and Ivory Coast (Sanchez, A. et al., 1996 PNAS USA 93: 3602-3607). These Ebolavirus subtypes have similar genetic organizations, for example, negative-strand RNA viras containing seven genes in a linear array. Structural gene products include, for example, envelope glycoprotein which is in two possible forms, a secreted soluble glycoprotein (ssGP) and a transmembrane glycoprotein (GP) generated by an RNA modification that allows viral entry. (Sanchez, A. et al., 1996 PNAS USA 93: 3602-3607). (008) It has been suggested that immunization may be useful for protection against Ebola infection and / or related disease, as there appears to be less nucleotide polymorphism among Ebola subtypes. than among other RNA viras (Sanchez, A. et al., 1996 PNAS USA 93: 3602-3607). Until recently, developments in preventive vaccines against filoviras have had variable results, in part because the conditions required for protective immune responses against filovirus infections are not sufficiently understood. (009) Currently, there are several vector-based antigen-based vaccine platforms that have demonstrated varying degrees of protection in nonhuman primates (HNP) exposed to high infectious doses of filoviras. Candidate vaccines in development are based on a variety of platform techniques, including replication-competent vectors (eg, viras of vesicular stomatitis, rabies viras, parafluenza viras); incompetent viruses for replication (adenoviras, modified vaccinia viras Ankara); protein subunits including viral-like particles expressed in bacterial cells, insect cells, mammalian cells, plant cells; DNA vaccines; and / or attenuated live or killed filoviras (Friedrich et al., 2012). EBOV GP glycoprotein is an important component of a vaccine that protects against exposure to the same EBOV species. The development of medical countermeasures for these viras is a high priority. (010) Replication deficient adenoviral vectors (rAd) are potent inducers of cellular immune responses and have therefore become useful vectors for gene-based vaccines, including for lentiviruses and filoviruses, as well as for other non-viral pathogens (Shiver et al., (2002) Nature 415 (6869): 331-5; (Hill et al., Hum Vaccine 6 (1): 78-83; Sullivan et al., (2000) Nature 408 (6812): 605-9, Sullivan et al., (2003) Nature 424 (6949): 681-4, Sullivan et al., (2006) PLoS Med 3 (6): el77, Radosevic et al. , (2007), Santra et al., (2009) Vaccine 27 (42): 5837-45.) Adenovirus-based vaccines offer several advantages as human vaccines, as they can be produced at elevated levels in humans. GMP conditions and have been shown to be safe and immunogenic in humans (Asmuth et al., J Infect Dis 201 (1): 132-41; Kibuuka et al., J Infect Dis 201 (4): 600-7 Koup et al., PLoS O No. 5 (2): e9015; Catanzaro et al., (2006) J Infect Dis 194 (12): 1638-49; Harro et al., (2009) Clin Vaccine Immunol 16 (9): 1285-92). Although most of the initial work on vaccines was done using rAd5, because of its clear ability to elicit broad-spectrum antibody responses and CD8 + T cell responses, pre-existing immunity against rAd5 in humans can limiting efficacy (Catanzaro, (2006) Cheng et al., (2007) PLoS Pathog 3 (2): e25, McCoy et al., (2007) J. Virol 81 (12): 6594-604.; Buchbinder et al., (2008) Lancet 372 (9653): 1881-93). This property could restrict the utility of rAd5 in the clinical applications of many vaccines that are currently under development, including those against Ebolavirus (EBOV). (011) Replication-defective adenoviral vectors, rAd26 and rAd35, derived from serotype 26 and adenovirus serotype 35, respectively, have the ability to escape pre-existing immunity against Ad5. rAd26 can be grown to high titers in Ad5-El complementing cell lines, suitable for the large-scale, clinical-grade manufacture of these vectors (Abbink et al., 2007), and it has been shown that this vector induced humoral and cellular immune responses in primer-boost vaccination strategies (Abbink et al., 2007, Liu et al., (2009) Nature 457 (7225): 87-91). The rAd35 vectors grow to high titers on cell lines suitable for the production of clinical grade vaccines (Havenga et al., (2006) J. Gen. Virol. 87 (Pt 8): 2135-43), and were formulated for injection as well as stable inhalable powder (Jin et al., Vaccine 28 (27): 4369-75). These vectors show efficient transduction of human dendritic cells (Gruijl et al., (2006) J. Immunol 177 (4): 2208-15 Lore et al., (2007) J. Immunol 179 (3): 1721-9), and thus have the ability to induce a high level of antigen intake and presentation. (012) The prevalence of immunity against human adenoviruses has also led to the consideration of simian adenoviruses as vectors. Simian adenoviruses are not known to be the cause of pathology or disease in humans, and the prevalence of antibodies against adenoviruses originating from chimpanzees is less than 5% in humans resident in the United States (Tatsis N et al., 2007 Mol. th .: The Journal of the American Society of Gene Therapy 15: 608-17). They exhibit hexon structures analogous to those of human adenoviruses (Bruna-Romero O. et al., 2001 Nat.Academic of Sciences of the USA 98: 11491-6). (013) Recombinant chimpanzee adenovirus serotype 3 (ChAd3 or ad3) is an adenovirus of subgroup C which has properties similar to those of serotype 5 human adenovirus (Ad5). Chimpanzee adenovirus ChAd63 is another simian vector that is used to prevent problems with pre-existing immunity against AdHu5 in humans. Both Adenovirus ChAd3 and Adenovirus cAd3 have been shown to be safe and immunogenic in human studies to evaluate candidate vaccines against hepatitis C virus (HCV) (Barnes et al. Novel Adenovirus-based Vaccines Induce Broad and Sustained T Cell Responses to HCV in Human Science Translational Medicine 2012; 4: 115ral) and Malaria (O'Hara et al., Clinical Assessment of a Recombinant Simian Adenovirus ChAd63: A Potent New Vaccinia Vector The Journal of Infectious Diseases 2012; 205: 772-81), respectively. (014) There is an unmet need for improved vaccines, eliciting immune responses against filovirs, particularly protective immunity against the most lethal Ebolaviruses, such as EBOV.
BRIEF SUMMARY OF THE INVENTION (015) It has been discovered in the invention that various primer-booster combinations of replication-incompetent vectors elicit an effective immune response against filovirus infection and / or disease. More specifically, a combination for primer-booster immunization of a first composition comprising at least one human adenoviral vector and a second composition comprising a simian adenoviral vector results in improved immune protection. (016) Accordingly, a general aspect of the invention relates to a combination of vaccine comprising: (i) a first composition comprising an immunologically effective amount of at least one human adenoviral vector comprising a first nucleic acid encoding a first antigenic protein; a filovirus, and a pharmaceutically acceptable carrier; and (ii) a second composition comprising an immunologically effective amount of a simian (e.g. chimpanzee) adenoviral vector comprising a second nucleic acid encoding a second filovirus antigenic protein, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (017) In a combination of vaccine according to one embodiment, the first antigenic protein and the second antigenic protein may be the same protein or different proteins. (018) Another general aspect of the invention relates to a method of inducing an immune response against a filovirus in a subject, comprising: a. administering to a first composition comprising an immunologically effective amount of a human adenoviral vector comprising a first nucleic acid encoding a first filovirus antigenic protein; and B. administering to a second composition comprising an immunologically effective amount of a simian (e.g. chimpanzee) adenoviral vector comprising a second nucleic acid encoding a second filovirus antigenic protein. steps (a) and (b) being performed in this order or in reverse order. (019) In a method according to one embodiment of the invention, the first antigenic protein and the second antigenic protein may be the same protein or different proteins. (020) In some embodiments, the first composition is used for priming said immune response and the second composition is used for boosting said immune response. In other embodiments, the second composition is used for priming said immune response and the first composition is used for boosting said immune response. (021) In one embodiment, the boosting composition is administered 1-15 weeks after the priming composition. In a preferred embodiment, the priming vaccination is performed at week 0, followed by a booster vaccination at week 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 15, 12, 13, 14, 15. In another preferred embodiment, priming vaccination is performed at week 0, followed by booster immunization at 6, 9, 12 or 24 months. More preferably, the initiating vaccination is carried out at week 0, followed by a booster vaccination at week 2, 4, 6, 8, 10 or 12. (022) Filovirus subtypes according to the invention can be any filovims. In preferred embodiments of the invention, filovims is an Ebola virus. In a preferred embodiment, the Ebola virus belongs to the Zaire subtype. (023) The antigenic protein encoded by the adenoviral vectors included in the first and second compositions may be any protein from any filovirus (e.g. Marburg or Ebola). In one embodiment, the antigenic protein is a glycoprotein (GP) or a nucleoprotein (NP) of a filovirus. In a preferred embodiment, the first antigenic protein comprises a GP comprising the amino acid sequence of SEQIDnO1 or a protein substantially similar to GP, such as a GP comprising the amino acid sequence of SEQ K) No. 3 In another embodiment, the second antigenic protein comprises a GP comprising the amino acid sequence of SEQ ID No. 1 or a protein substantially similar to GP, such as a GP comprising the amino acid sequence of SEQ ID No. 3. In yet another embodiment, each of the first and second antigenic proteins comprises a GP comprising the amino acid sequence of SEQ ID No. 1 or a protein substantially similar to GP, such as GP comprising the amino acid sequence of SEQ ID No. 3. (024) It is contemplated that the methods, vaccines and compositions described herein can be made into a kit. For example, in one embodiment, the invention may include a kit comprising: (i) a first composition comprising an immunologically effective amount of at least one human adenoviral vector comprising a first nucleic acid encoding a first antigenic protein of a filovirus, and a pharmaceutically acceptable carrier; and (ii) a second composition comprising an immunologically effective amount of a simian (e.g. chimpanzee) adenoviral vector comprising a second nucleic acid encoding a second filovirus antigenic protein, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (025) In said kit, the first antigenic protein and the second antigenic protein may be the same protein or different proteins. (026) In a preferred embodiment, the human adenoviral vector comprised in the first composition of the vaccine combination, the method or kit is a rAd26 or rAd35 vector, more preferably a rAd26 vector. In another preferred embodiment, the simian adenoviral vector in the second composition of the vaccine combination, the method or kit is an adenoviral chimpanzee vector, such as a ChAd3 vector. (027) Accordingly, in one embodiment, the invention relates to a combination vaccine, kit, or method, wherein a first composition comprises an immunologically effective amount of at least one human adenoviral vector, and wherein a second composition comprises an immunologically effective amount of a simian adenoviral vector, such as an adenoviral chimpanzee vector. The first composition comprises an immunologically effective amount of at least one human adenoviral vector comprising a nucleic acid encoding an antigenic protein of a filovirus, and the second composition comprises an immunologically effective amount of a simian adenoviral vector comprising a nucleic acid encoding a antigenic protein of a filovirus, which may be the same as or different from the antigenic protein encoded by said at least one human adenoviral vector in the first composition.
BRIEF DESCRIPTION OF THE DRAWINGS (028) The reading of the foregoing summary, as well as the following detailed description of the invention will be better understood in conjunction with the accompanying drawings. It should be understood that the invention is not limited to the specific embodiments shown in the drawings. (029) In these drawings: (030) FIG. 1 is a graph showing the specific humoral immune response of GP-EPOV, evaluated by an enzyme linked immunosorbent assay (ELISA), expressed in ELISA units per ml, on test samples taken from G1 and G3 group subjects. at weeks 1 to 21 of the test: and (031) FIG. 2 is a graph showing the GP-EPOV specific humoral immune response, evaluated by enzyme linked immunosorbent assay (ELISA), expressed in ELISA units per ml, on test samples taken from G2 and G4 group subjects. , at weeks 1 to 21 of the test.
DETAILED DESCRIPTION OF THE INVENTION (032) Various publications, various articles, and various patents are cited and described in the background and throughout the description; each of these references is incorporated in its entirety here by reference. The review of documents, acts, materials, devices, articles or the like that has been included in the description is for the purpose of providing context for the invention. Π should be understood that such an examination is not an admission that some or all of these materials are part of the state of the art with respect to any inventions described or claimed. (033) Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. Otherwise, some terms used herein have the meanings specified in the description. All patents, all published patent applications and publications cited herein are incorporated by reference as fully referenced herein. Π must be understood that as used herein and in the appended claims, the singular forms "a", "an" and "the" or "the" include the reference to the plural, unless expressly stated otherwise by the context. (034) Unless otherwise indicated, the term "at least" preceding a series of elements shall be understood to refer to each element in the series. Those skilled in the art will recognize, or be able to determine, without having to use more than one routine experiment, many equivalents of the particular embodiments of the invention described herein. Such equivalents should be understood to be encompassed by the invention. (035) Throughout the present description and the appended claims, unless otherwise indicated by the context, the term "include", and its variants such as "includes" and "comprising" must be understood to mean the inclusion of a stated integer or step or of a group of integers or steps, but not the exclusion of any other integer or any other step or any group of integers or steps. When used herein, the term "comprising" may be replaced by the term "containing" or "including" or sometimes when used herein, by the term "having". When used herein, the term "consisting of" excludes any element, step or component not specified in the claimed element. When used herein, the term "substantially consisting of" does not exclude materials or steps that do not materially affect the basic and novel features of the claim. In each case herein, any of the terms "comprising", consisting essentially of "and consisting of" may be replaced by either of the other two terms. (036) As used herein, the connective term "and / or" between many of the stated elements should be understood to encompass both the individual options and the combined option. For example, when two elements are joined by "and / or", a first option refers to the applicability of the first element without the second. A second option relates to the applicability of the second element without the first. A third option relates to the application of the first and second elements together. Any of these options must be understood as falling within this meaning, and therefore as meeting the requirement of the term "and / or" as used herein. Concomitant applicability of more than one of the options must also be understood as meaning, and therefore as satisfying the requirement of "and / or". (037) As used herein, a "subject" means an animal turn, preferably a mammal, most preferably a human being, who will be or has been vaccinated by a method according to one embodiment of the invention. As used herein, the term "mammal" encompasses any mammal. Examples of mammals include, but are not limited to, cattle, horses, sheep, swine, cats, dogs, mice, rats, rabbits, guinea pigs, monkeys, humans, etc., more particularly preferably a To be human. (038) As used herein, the term "protective immunity" or "protective immune response" means that the vaccinated subject is capable of controlling infection with the pathogen against which the vaccination has been carried out. Usually, the subject who has developed a "protective immune response" has only mild to moderate clinical symptoms or none at all. Usually, a subject having a "protective immune response" or "protective immunity" against a particular agent will not die as a result of infection with the agent. (039) A "capsidic protein" refers to a protein present on the capsid of an adenovirus (eg Ad26 or Ad35) that is involved in the determination of the serotype and / or tropism of a particular adenovirus. The capsidic proteins typically include fibrous, penton and / or hexon proteins. As used herein, an "Ad26 capsidic protein" or "Ad35 capsid protein" may be, for example, a chimeric core protein that includes at least a portion of an ad26 capsid protein. or Ad35. In some embodiments, the capsid protein is an integral capsid protein of Ad26 or Ad35. In some embodiments, the hexon, penton, or fiber is Ad26 or Ad35. (040) The term "adjuvant" is defined as one or more substances that cause stimulation of the immune system. In this context, an adjuvant is used to increase the immune response to the adenoviral vectors of the invention. (041) The term "identical" or "identity" in percent, in the context of two or more nucleic acid or polypeptide sequences (eg, filovirus glycoproteins and polynucleotides encoding them), refers to to two or more sequences or subsequences that are identical or have a specified percentage of amino acid residues or nucleotides that are identical, when compared and aligned for maximum match, as measured using the one of the sequence comparison algorithms hereinafter or by visual examination. (042) For the comparison of sequences, normally a sequence acts as a reference sequence, to which the sequences to be tested are compared. When using a sequence comparison algorithm, the reference and test sequences are entered into a computer, sequence coordinates are defined, if necessary, and the parameters of the sequence algorithm program are defined. The sequence comparison algorithm then calculates the sequence identity in percent for the test sequence (s), relative to the reference sequence, based on the defined program parameters. (043) An optimal alignment of sequences for comparison can be performed, for example, by the local homology algorithm of Smith and Waterman, Adv. Appl. Math. 2: 482 (1981), by the homology alignment algorithm of Needleman and Wunsch, J. Mol. Biol. 48: 443 (1970), by the similarity search method of Pearson and Lipman, Proc. Nat'l. Acad. Sci. USA 85: 2444 (1988), by computerized execution of these algorithms (GAP, BESFIT, FASTA and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by examination. visual (see generally Current Protocols in Molecular Biology, FM Ausubel et al., editors, Current Protocols, a partnership between Greene Publishing Associates, Inc. and John Wiley & Sons, Inc., (Supplement 1995) (Ausubel)). (044) Examples of algorithms that are suitable for determining percent sequence identity or sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al. (1990) J. Mol. Biol. 215: 403-410 and Altschuel et al. (1977) Nucleic Acids Res. 25: 3389-3402, respectively. The program for running BLAST analyzes is publicly available through the National Center for Biotechnology Information. This algorithm firstly comprises the identification of pairs of high scoring pairs (HSP) by identification of short words of length W in the sequence in question, which correspond to or satisfy a certain threshold score T of positive value when aligned with a word of the same length in a database sequence. T is considered to be the neighboring word score threshold (Altschul et al., Supra). These successful sets of initial neighbor words act like seeds to initiate searches to find longer HSPs containing them. Successful sets of words are then extended in both directions along each sequence, as far as it is possible to increase the cumulative alignment score. (045) Cumulative scores are calculated using, for the nucleotide sequences, the parameters M (reward score for a pair of corresponding residues, always> 0) and N (penalty score for residues that do not match, always < 0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. The extension of the successful sets of words in each direction is interrupted when: the cumulative alignment score decreases by the amount X from its maximum value reached: the cumulative score reaches zero or less, due to the accumulation one or more negative-scoring residue alignments; or the end of one or the other sequence is reached. The parameters of the BLAST algorithm W, T and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as default values a word length (W) of 11, a prediction (E) of 10, M = 5, N = 4, and a comparison of two strands. For amino acid sequences, the BLASTP program uses as default values a word length (W) of 3, a prediction (E) of 10, and the score matrix BLOSUM62 (see Henikoff and Henikoff, Proc Nat '. IAcad, USA 89: 10915 (1989)]. (046) In addition to calculating percent sequence identity, the BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, for example, Karlin et al. Altschul, Proc Natl Acad Sci USA 90: 5873-5787 (1993)) A measure of the similarity provided by the BLAST algorithm is the lowest probability of the sum (P (N)), which gives an indication of the probability that a match between two nucleotide or amino acid sequences occurs by chance For example, a nucleic acid is considered to be similar to a reference sequence if the lowest probability of the sum in a comparison nucleic acid test with nuc acid The reference level is less than about 0.1, more preferably less than about 0.01, and most preferably less than about 0.001. (047) Another indication that two nucleic acid or polypeptide sequences are essentially identical is that the polypeptide encoded by the first nucleic acid immunologically reacts at the same time with the polypeptide encoded by the second nucleic acid, as described below. Thus, a polypeptide is normally substantially identical to a second polypeptide, for example, when the two peptides differ only in conservative replacements. Another indication that two nucleic acid sequences are essentially identical is that the two molecules hybridize to one another under stringent conditions, as described below. (048) The term "substantially similar" in the context of the filovirus antigenic proteins of the invention indicates that a polypeptide comprises a sequence having a sequence identity percentage of at least 90%, preferably from minus 95%, with the reference sequence. The percentage of sequence identity is determined by comparing two optimally aligned sequences in a comparison window, the portion of the polynucleotide sequence in the comparison window possibly including additions or deletions (i.e. holes) in comparison with the reference sequence (which does not include additions or deletions) for the optimal alignment of the two sequences. The percentage is calculated by determining the number of positions in which the nucleic acid base or amino acid residue appears in both sequences to obtain the number of matched positions, and then dividing the number of matched positions by the total number of positions. in the comparison window and multiplying the result by 100 to get the percentage of sequence identity. (049) It is discovered in the invention that various primer-abortive combinations of replication-incompetent vectors elicit an effective immune response against filovirus infection and / or disease. More specifically, a primer-booster combination of a first composition comprising at least one human adenoviral vector and a second composition comprising a simian adenoviral vector, such as an adenoviral chimpanzee vector, gives rise to an improved immune response.
Antigenic Filovirus Proteins (050) Ebola viruses are filoviruses associated with outbreaks of highly lethal haemorrhagic fever in humans and primates in North America, Europe and Africa (Peters, C. J. et al., In Fields). Virology, Fields, BN et al., Eds., 1161-1176, Philadelphia, Lippincott-Raven, 1996, Peters, CJ et al., 1994 Semin Virol 5: 147-154). Although several subtypes have been defined, the genetic organization of these viruses is similar, each containing seven genes in a linear arrangement. Among the viral proteins, the envelope glycoprotein is found in two different forms, a 50-70 kilodalton (kDa) secreted protein (sGP) and a 130 kDa transmembrane glycoprotein (GP), generated by RNA modification that allows viral entry (Peters, CJ, et al., Virology, Fields, BN et al., eds., 1161-1176, Philadelphia, Lippincott-Raven, 1996, Sanchez, A. et al., 1996 PNAS USA 93: 3602). -3607). Other structural gene products include nucleoprotein (NP), VP24 and VP40 template proteins, presumed VP30 and VP35 non-structural proteins, and viral polymerase (summarized in the review article by Peters, C.J. in Fields Virology, Fields, BN et al., eds., 1161-1176, Philadelphia, Lippincott-Raven, 1996). (051) The nucleic acid molecules included in the adenoviral vectors can encode structural gene products of any species of filovirus (Marburg and / or Ebola), such as Zaire subtypes (type species, also referred to as here by EBOV), Sudan, Reston, Bundibugyo and Ivory Coast, including but not limited to those described in WO 2003/028632, the contents of which are hereby incorporated in their entirety by reference. The nucleic acid molecules included in the adenoviral vectors can encode structural gene products of filovirus species involved in a particular outbreak, such as species reported to be related to the 2014/2015 outbreak or any future outbreak. . (052) The adenoviral vectors of the invention can be used to express antigenic proteins comprising an antigenic determinant of a wide variety of filovirus antigens. In a typical and preferred embodiment, the vectors of the invention comprise a nucleic acid encoding the transmembrane form of the viral glycoprotein (GP). In other embodiments, the vectors of the invention may encode the secreted form of the viral glycoprotein (ssGP), or the viral nucleoprotein (NP). (053) It will be clear to those skilled in the art that the nucleic acid molecules encoding the filovirus antigenic protein may be modified, for example, the nucleic acid molecules indicated herein may be mutated, as long as the modified expressed protein elicit an immune response against a pathogen or disease. Thus, as used herein, the term "antigenic protein" or "filovirus protein" refers to a protein that comprises at least one antigenic determinant of a filovirus protein, such as those described above. The term encompasses filovirus glycoproteins (i.e. gene products of gene 4 or filovirus GP gene) or filovirus nucleoprotein as well as recombinant proteins that include one or more determinants of filovirus nucleoproteins or glycoproteins. . The term antigenic proteins also encompasses antigenic proteins that are substantially similar to naturally occurring filovirus proteins. (054) In some embodiments, the protein may be mutated, so as to be less toxic to the cells (see for example WO / 2006/037038), the contents of which are incorporated in their entirety herein by reference, or may be expressed to an increased or decreased degree in the cells. In a preferred embodiment, nucleic acid molecules encoding GP, ssGP and NP of the Ebola Zaire virus strain are included in the first and second compositions of the vaccine combination. Preferably, the vector in the first composition of the vaccine combination encodes a GP having the amino acid sequence of SEQ ID No. 1, or a protein substantially similar to SEQ ID No. 1, such as a protein comprising the sequence of amino acids of SEQ ID No. 3, preferably the protein is capable of inducing an immune response against a GP having the amino acid sequence of SEQ ID No. 1. In another preferred embodiment, the vector in the second composition of the vaccine combination encodes a GP having the amino acid sequence of SEQ ID No. 1, or an amino acid sequence substantially similar to SEQ ID No. 1, such as a protein comprising the amino acid sequence of SEQ ID No. 3, preferably the protein is capable of inducing an immune response against a GP having the amino acid sequence of SEQ ID No. 1. In a more particularly preferred embodiment, the in the first composition of the vaccine combination encodes a GP having the amino acid sequence of SEQ ID No. 1 or SEQ ID No. 3, and the vector in the second composition of the vaccine combination codes a GP having the amino acid sequence of SEQ ID No. 1 or SEQ ID No. 3.
Human adenoviral vectors (055) An adenoviral vector according to the invention is derived from an adenovirus which belongs to the family of Adenoviridae and preferably is an adenovirus which belongs to the genus Mastadenovirus. In a preferred embodiment, the adenoviral vector of the first composition is derived from a human adenovirus (HAdV or
AdHu; in the present description is meant as human adenovirus if it relates to Ad, without indication of species, for example, the abbreviated name "Ad5" has the same meaning as HAdV5, which is the serotype of human adenovirus 5). (056) Most of the extensive studies have been done using human adenoviruses. In certain embodiments, the recombinant adenovirus is based on a human adenovirus serotype 5, 11, 26, 34, 35, 48, 49 or 50. According to a particularly preferred embodiment of the invention, an adenoviral vector is derived from a human adenovirus of one of serotypes 26 and 35. An advantage of these serotypes is low seroprevalence and / or the existence of low levels of pre-existing neutralizing antibodies in the human population. The preparation of rAd26 vectors is described, for example, in WO 2007/104792 and in Abbink et al., (2007) Virol 81 (9): 4654-63, the contents of which are incorporated herein by reference. Genomic sequences taken as examples of Ad26 are found in GenBank, accession number EF 153474 and in SEQ ID No. 1 of WO 2007/104792, which are incorporated in their entirety herein by reference. The preparation of rAd35 vectors is described, for example, in US Pat. No. 7,270,811, WO 00/70071 and Vogels et al., (2003) J. Virol 77 (15): 8263-71, all of which are incorporated herein by reference. incorporated in their entirety here by reference. Genomic sequences taken from Ad35 examples are found in GenBank, record number AC_000019 and in FIG. 6 of WO 00/70071, which are incorporated in their entirety herein by reference. (057) In a preferred embodiment, human adenoviral vectors comprise capsid proteins from two rare serotypes: Ad26 and Ad35. In one embodiment, the vector is a rAd26 or rAd35 virus. (058) The vectors that can be used in the invention thus comprise a capsid protein of Ad26 or Ad35 (for example a fibrous protein, with pentons or hexons). Those skilled in the art will recognize that in the vectors of the invention it is not necessary to use a complete AD26 or Ad35 capsid protein. Thus, chimeric capsid proteins that include at least a portion of an Ad26 or Ad35 capsid protein can be used in the vectors of the invention. The vectors of the invention may also comprise capsid proteins in which the fibrous, penton and hexon proteins are each derived from a different serotype, insofar as at least one capsidic protein is derived from Ad26 or Ad35. In preferred embodiments, the fibrous, penton and hexon proteins are each derived from Ad26 or each Ad35. (059) It will be clear to one of ordinary skill in the art that elements from multiple serotypes can be combined into a single recombinant adenoviral vector, for example a human adenovirus or a chimpanzee. It is therefore possible to produce a chimeric adenoviral vector combining desirable properties of different serotypes. Thus, in certain embodiments, a chimeric human adenoviral vector of the invention could combine the absence of pre-existing immunity of Ad26 and Ad35 serotypes with characteristics such as temperature stability, assembly, anchorage, production yield, redirected or improved infection, stability of the DNA in the target cell and the like. (060) In some embodiments, the recombinant human adenoviral vector for use in the invention is derived primarily or entirely from Ad35 or Ad26 (i.e., the vector is rAd35 or rAd26). In some embodiments, the human adenovirus is defective for replication, for example by containing a deletion in the E1 region of the genome. For human adenoviruses taken as examples of the invention, which are derived from Ad26 or Ad35, it is characteristic to replace the E4-orf6 coding sequence of the adenovirus with the E4-orf6 sequence of a subgenic adenovirus. human group C such as Ad5. This allows the propagation of such adenoviruses in well-known complementing cell lines that express the Ad5 E1 genes, such as 293 cells, PER.C6 cells and the like (see for example Havenga et al, 2006, J Gen Virol 87: 2135-43; WO 03/104467, which are incorporated in their entirety herein by reference). In some embodiments, the adenovirus is a human serotype 35 adenovirus, with a deletion in the E1 region in which the nucleic acid encoding the antigen has been cloned, and having an E4 orf6 region of Ad5. In some embodiments, the adenovirus is a human serotype 26 adenovirus, having a deletion in the E1 region in which the nucleic acid encoding the antigen has been cloned, and having an E4 orf6 region of Ad5. For adenovirus Ad35, it is characteristic to keep the 3 'end of the El B 55K open reading frame in the adenovirus, for example the 166 bp directly upstream of the open reading frame pIX or a fragment comprising them, such as that a 243 bp fragment directly upstream of the pIX initiation codon, labeled at the 5 'end by a Bsu361 restriction site, since this increases the stability of the adenovirus because the pIX gene promoter is found partly in this region (see, for example, Havenga et al, 2006, cit., WO 2004/001032). (061) A human adenoviral vector can be prepared using methods known in the art, in view of the present disclosure. The preparation of rAd26 vectors is described, for example, in WO 2007/104792 and in Abbink et al., (2007) Virol 81 (9): 4654-63. Genomic sequences taken as examples of Ad26 are found in GenBank, accession number EF 153474, and in SEQ ID NO: 1 of WO 2007/104792. The preparation of rAd35 vectors is described, for example, in US Pat. No. 7,270,811 and in Vogels et al. (2003) J. Virol 77 (15): 8263-71. A genomic sequence taken as an example of Ad35 is found in GenBank, record number AC_000019. (062) In one embodiment, the vectors usable for the invention include those described in WO 2012/082918, the disclosure of which is incorporated herein by reference.
Simian adenoviral vectors (063) The simian adenoviral vectors according to the present invention are derived from adenoviruses originating from simians, which, as used herein, include monkeys and anthropoids, but exclude humans. Examples of simians include, but are not limited to, chimpanzees, gorillas, orangutans, etc. Simian adenoviruses generally have low seroprevalence and / or low levels of pre-existing neutralizing antibodies in the human population, and many studies that have used simian adenoviral vectors, such as adenoviral chimpanzee vectors, have been reported (e.g., US 6083716, WO 2005/071093, WO 2010/086189, WO 2010085984, Farina et al, 2001, Virol 75: 11603-13, Cohen et al, 2002, J Gen Virol 83: 151-55; et al., 2006, Virology 346: 394-401, Tatsis et al., 2007, Molecular Therapy 15: 608-17, see also the review article of Bangari and Mittal, 2006, Vaccine 24: 849-62; the review article by Lasaro and Ertl, 2009, Mol Ther 17: 1333-39, all of which are incorporated herein by reference). Consequently, in other preferred embodiments, the recombinant adenoviral vector according to the invention is based on a simian adenovirus, for example a chimpanzee or bonobo adenovirus. In some embodiments, the recombinant adenovirus is based on a chimpanzee adenovirus type 1, 3, 7, 8, 21, 22, 23, 24, 25, 26, 27.1, 28.1, 29, 30, 31.1, 32 , 33, 34, 35.1, 36, 37.2, 39, 40.1, 41.1, 42.1, 43, 44, 45, 46, 48, 49, 50, 67 or SA7P. (064) In simian adenoviral vectors, such as adenoviral chimpanzee vectors, it is possible to suppress the El locus in order to make the viruses defective for replication and to allow transcomplementation on an El AdHU5 complementing cell line (Farina SF, et al., 2001 J Virol 75: 11603-13). A further interesting observation is that the lack of sequence homology between AdHu5 and simian adenoviruses at the level of the E1-adjacent sequence prevents homologous recombination and production of replication competent viruses (Tatsis N, et al., 2006 Gene Ther 13: 421-9). In some embodiments, the recombinant adenoviral vector is based on type 3 or 63 chimpanzee adenovirus. (065) In a preferred embodiment, the simian adenoviral vector of the second composition is an adenoviral chimpanzee vector (ChAdV) or AdCh). (066) In a more particularly preferred embodiment, the adenoviral chimpanzee vector of the second composition is ChAdV3. Recombinant chimpanzee adenovirus serotype 3 (ChAd3 or ad3) is an adenovirus of subgroup C which has similar properties to human adenovirus serotype 5 (Ad5). In studies in human subjects to evaluate candidate vaccines against hepatitis C virus (HCV), it has been shown that ChAd3 is safe and immunogenic (Barnes E, et al., 2012 Science translational medicine 4: 115al ). It has been reported that ChAd3 vaccines are capable of inducing an immune response comparable to a human Ad5 vector vaccine. See for example
Perazzi D, et al. 2009 Vaccine 27: 1293-300 and Quinn KM, et al. 2013 J Immunol 190: 2720-35; WO 2005/071093; WO2011 / 0130627, etc. (067) It is possible to produce a vector usable in the invention by using a nucleic acid comprising the recombinant adenoviral genome (for example, a vector of plasmid, cosmid or baculovirus type). The invention thus provides isolated nucleic acid molecules that encode the adenoviral vectors of the invention. The nucleic acid molecules of the invention may be in the form of RNA or in the form of DNA obtained by cloning or produced by synthesis. DNA can be single-stranded or double-stranded. (068) The human and simian adenoviral vectors that can be used for the invention are normally defective for replication. In these embodiments, the virus is defective for deletion or inactivation replication of regions that are critical for virus replication, such as the EL region. Regions can be essentially deleted or inactivated, for example, by insertion of the gene. interest (usually linked to a promoter). In some embodiments, the vectors of the invention may contain deletions in other regions, such as E2, E3 or E4 regions, or heterologous gene insertions linked to a promoter. For E2 and / or E4 mutated adenoviruses, E2 and / or E4 complementing cell lines are generally used to generate recombinant adenoviruses. Mutations in the E3 region of the adenovirus do not need to be complemented by the cell line because E3 is not required for replication. (069) A packaging cell line is normally used to produce a sufficient amount of adenoviral vectors of the invention. A packaging cell is a cell that includes genes that have been deleted or inactivated in a defective vector for replication, allowing the virus to replicate in the cell. Suitable cell lines include, for example, Procell-92, PER.C6, 911, 293 and El A549. (070) As indicated above, a wide variety of filovirus glycoproteins can be expressed in vectors. If necessary, the heterologous gene encoding the filovirus glycoproteins can be codon optimized to ensure adequate expression in the treated (eg, human) host. Codon optimization is a widely applied technique in the field. Normally, the heterologous gene is cloned in the E1 region and / or the E3 region of the human adenoviral genome. (071) The heterologous filovirus gene may be under the control of one (i.e., operably linked to) adenovirus-derived promoter (e.g., Major
Late Promoter) or may be under the control of a heterologous promoter. Examples of suitable heterologous promoters include, but are not limited to, the CMV promoter, for example the hCMV promoter and the RSV promoter. Preferably, the promoter is located upstream of the heterologous gene of interest within an expression cassette. (072) As indicated above, the human and simian adenoviral vectors that can be used for the invention can comprise a wide variety of filovirus glycoproteins known to those skilled in the art. (073) In a preferred embodiment of the invention, the vector (s) of rAd and ChAdV comprises / comprise one or more GPs of Ebola Zaire virus (EVOP) or GP substantially similar thereto.
Immunogenic combination (074) A vaccination strategy to achieve protective immunity in most recipients with a single vaccination would be desirable in an outbreak situation. Immunization strategies to achieve sustainable protective immunity would be desirable for populations in areas of the world where outbreaks occur sporadically. Optimally, a strategy would meet both needs, but a different strategy may be required for the rapid immunity that is needed for long-lasting immunity. (075) A general aspect of the invention is a vaccine or immunogenic combination comprising: (i) a first composition comprising an immunologically effective amount of at least one human adenoviral vector comprising a first nucleic acid encoding a first antigenic protein of a filovirus, and a pharmaceutically acceptable carrier; and (ii) a second composition comprising an immunologically effective amount of a simian adenoviral vector comprising a second nucleic acid encoding a second filovirus antigenic protein, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (076) Another aspect of the invention is a vaccine or immunogenic combination for inducing protective immunity against filovirus disease and / or infection, comprising: (i) a first composition comprising an immunologically effective amount at least one human adenoviral vector comprising a first nucleic acid encoding a first antigenic protein of a filovirus, and a pharmaceutically acceptable carrier; and (ii) a second composition comprising an immunologically effective amount of a simian adenoviral vector comprising a second nucleic acid encoding a second filovirus antigenic protein, the second nucleic acid being the same as the first nucleic acid or different from the first nucleic acid, and a pharmaceutically acceptable vehicle; one of the compositions being a priming composition and the other composition being a booster composition. (077) In a preferred embodiment of the invention, a combination of vaccine comprises: (i) a first composition comprising an immunologically effective amount of rAd26 or rAd35 vector comprising a first nucleic acid encoding a first antigenic protein having the amino acid which is at least 90% identical to SEQ ID NO: 1 and a pharmaceutically acceptable carrier; and (ii) a second composition comprising an immunologically effective amount of ChAd3 vector comprising a second nucleic acid encoding a second antigenic protein having the amino acid sequence which is at least 90% identical to SEQ ID NO: 1, and a pharmaceutically acceptable vehicle; one of the compositions being a priming composition and the other composition being a booster composition. (078) In a more particularly preferred embodiment of the invention, a combination of vaccine comprises: (i) a first composition comprising an immunologically effective amount of rAd26 vector comprising a first nucleic acid encoding an antigenic protein having the sequence of amino acids of SEQ ID No. 1 or SEQ ID No. 3, and a pharmaceutically acceptable carrier; and (ii) a second composition comprising an immunologically effective amount of ChAd3 vector comprising a second nucleic acid encoding the antigenic protein having the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (079) In another more particularly preferred embodiment of the invention, a combination of vaccine comprises: (i) a first composition comprising an immunologically effective amount of rAd26 vector comprising a first nucleic acid encoding an antigenic protein having the sequence of amino acids of SEQ ID NO: 1, and a pharmaceutically acceptable carrier; and (ii) a second composition comprising an immunologically effective amount of ChAd3 vector comprising a second nucleic acid encoding the antigenic protein having the amino acid sequence of SEQ ID NO: 3, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (080) In another preferred embodiment of the invention, a combination of vaccine comprises: (i) a first composition comprising an immunologically effective amount of rAd26 vector comprising the nucleotide sequence of SEQ ID No. 2, and a vehicle pharmaceutically acceptable; and (ii) a second composition comprising an immunologically effective amount of ChAd3 vector comprising the nucleotide sequence of SEQ ID NO: 2, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (081) Another aspect of the invention relates to a kit comprising: (i) a first composition comprising an immunologically effective amount of at least one human adenoviral vector comprising a first nucleic acid encoding a first antigenic protein of a filovirus, and a pharmaceutically acceptable vehicle; and (ii) a second composition comprising an immunologically effective amount of a simian adenoviral vector comprising a second nucleic acid encoding a second antigenic protein, the second nucleic acid being the same as the first nucleic acid or different from the first nucleic acid, and a pharmaceutically acceptable vehicle; one of the compositions being a priming composition and the other composition being a booster composition.
Immunogenic Composition (082) The immunogenic compositions are compositions which comprise an immunologically effective amount of purified or partially purified human or simian (e.g., chimpanzee) adenoviral vectors for use in the invention. The compositions may be formulated as a vaccine (also referred to as an "immunogenic composition") according to methods well known in the art. Such compositions may include adjuvants to enhance immune responses. The optimum ratios of each component in the formulation can be determined by techniques well known to those skilled in the art, in view of the present disclosure. (083) The immunogenic compositions according to the embodiments of the present invention may be prepared using methods known to those skilled in the art in view of the present description. Liquid pharmaceutical compositions generally include a liquid carrier such as water, petroleum, vegetable or animal oils, mineral oil or synthetic oil. It may also include saline solution, a solution of glucose or other saccharide, or glycols such as ethylene glycol, propylene glycol or polyethylene glycol. (084) Compositions of the invention may include other filovirus antigens, or priming or booster inoculations may include other antigens. Other antigens used in combination with the adenoviral vectors of the invention may be, for example, filovirus antigens and nucleic acids expressing them. (085) The immunogenic compositions useful in the invention may comprise adjuvants. Adjuvants suitable for co-administration according to the invention should be adjuvants which are potentially safe, well tolerated and effective in humans, including QS-21, Detox-PC, MPL-SE, MoGM-CSF, TiterMax-G, CRL-1005, GERBU, TERAMIDE, PSC97B, MWO, PG-026, GSK-I, AS01, ASO3, ASO4, AS15, GcMAF, B-alethine, MPC-026, Adjuvax, CpG ODN, betafectin, alum and MF59. (086) Other adjuvants that can be administered include lectins, growth factors, cytokines and lymphokines such as interferon alpha, interferon gamma, platelet derived growth factor (PDGF), factor stimulating granulocyte formation (gCSF), granulocyte and macrophage colony stimulating factor (gMCSF), tumor necrosis factor (TNF), epidermal growth factor (EGF), IL-I, IL-2, IL-4, IL-6, IL-8, IL-10 and IL-12 or the nucleic acids that encode them. (087) The compositions of the invention may comprise an excipient, carrier, buffer, stabilizer, pharmaceutically acceptable, or other substances well known to those skilled in the art. These substances should be non-toxic and should not interfere with the effectiveness of the active component. The first and second compositions may comprise the same pharmaceutically acceptable carrier or different pharmaceutically acceptable carriers. The precise nature of the vehicle or other substance may depend on the route of administration, for example the intramuscular, subcutaneous, oral, intravenous, cutaneous, intra-mucosal (eg intestine), intranasal or intraperitoneal routes.
Method for Inducing Protective Immunity Against Filovirus Disease and / or Infection (088) Another general aspect of the invention relates to a method of inducing an immune response against a filovirus in a subject. The method comprises: a. administering to a first composition comprising an immunologically effective amount of a human adenoviral vector comprising a first nucleic acid encoding a first antigenic protein of a filovirus; and B. administering to a second composition comprising an immunologically effective amount of a simian adenoviral (e.g., chimpanzee) vector comprising a second nucleic acid encoding a second antigenic protein of a filovirus, the second antigenic protein being the same as or different from the first antigenic protein, wherein steps (a) and (b) are performed in that order or in the reverse order. (089) In some embodiments, the first composition is used for priming said immune response and the second composition is used for boosting said immune response. In other embodiments, the second composition is used for priming said immune response and the first composition is used for boosting said immune response. (090) Any of the immunogenic compositions according to the embodiments of the invention including, but not limited to, those described herein, may be used in the methods of the invention. (091) In one embodiment of the described methods, an adenoviral vector is used to prime the immune response and another adenoviral vector is used to boost the immune response, about 1-15 weeks after the priming vaccination, by for example, the priming vaccination can be administered 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 weeks after the priming vaccination. Additional priming compositions may be administered weeks or months after the initial booster administration, for example, about 1, 2 or 3 weeks or 4 weeks, or 8 weeks, or 16 weeks, or 20 weeks, or 24 weeks. weeks, or 28 weeks, or 32 weeks, or 36 weeks, or one or two years after the initial recall. (092) In a preferred embodiment, the human adenoviral vector that is used according to the present invention includes a rAd26 or rAd35 vector and the simian adenoviral vector includes an adenoviral chimpanzee vector, such as a ChAd3 vector. In an exemplary embodiment, a rAd26 or rAd35 vector is used to prime the immune response, and a ChAd3 vector for boosting the immune response, or vice versa. (093) The antigens in the respective priming and booster compositions need not be identical, but should preferably have similar antigenic determinants in common, or be substantially similar to each other. (094) Administration of the immunogenic compositions comprising the vectors is normally performed intramuscularly or subcutaneously. However, other modes of administration, such as intravenous, cutaneous, intradermal or nasal, can also be considered. The intramuscular administration of the immunogenic compositions can be carried out using a needle to inject a suspension of the adenoviral virus. Another possibility is to use a needleless injection device to administer the composition (using, for example, Biojector) or a freeze-dried powder containing the vaccine. (095) For intravenous, cutaneous or subcutaneous injection, or injection at the site of disease, the vector will be in the form of a parenterally acceptable aqueous solution, which is pyrogen-free and has a pH, isotonicity and stability. Those skilled in the art will be able to prepare suitable solutions using, for example, isotonic vehicles such as Sodium Chloride Injection, Ringer's Injection, Lactated Ringer's Injection. Preservatives, stabilizers, buffers, antioxidants and / or other additives may be included as needed. It is also possible to use a slow release formulation. (096) Normally, the administration will have a prophylactic goal to elicit an immune response against a filovirus antigen prior to infection or the onset of symptoms. Diseases and conditions that can be treated or prevented according to the invention include those in which an immune response may play a protective or therapeutic role. In other embodiments, the adenoviral vectors may be administered for post-exposure prophylaxis. (097) Immunogenic compositions containing human or simian adenoviral vectors (eg, chimpanzee) are administered to a subject, giving rise to an anti-filovirus immune response in the subject. An amount of a composition that is sufficient to induce a detectable immune response is defined as an "immunologically effective dose". As shown below, the immunogenic compositions of the invention induce a humoral immune response as well as a cellular immune response. In a typical embodiment, the immune response is a protective immune response. (098) The actual amount administered, and the rate and timing of administration will depend on the nature and severity of the condition being treated. The treatment prescription, eg dosing decisions, etc., will be under the responsibility of general practitioners and other medical doctors, or, in a veterinary context, a veterinary surgeon, and will normally take into account condition to be treated, the condition of the individual patient, the site of administration, the mode of administration and other factors known to practitioners. Examples of the techniques and protocols mentioned above can be found in Remington's Pharmaceutical Sciences, 16th Edition, Osol, A. Editor, 1980. (099) After the production of adenoviral vectors and the optional formulation of such particles into compositions, the vectors may be administered to an individual, particularly a human or other primate. The administration may be to human subjects, or to another mammal, for example a mouse, a rat, a hamster, a guinea pig, a rabbit, a sheep, a goat, a pig, a horse, a bovine, a donkey, a monkey, a dog or a cat. Administration to a non-human mammal is not necessarily for a therapeutic purpose, but may be for use in an experimental setting, for example, in the study of mechanisms of immune responses to adenoviral vectors. (0100) In an exemplary scheme, the human adenoviral vector is administered (e.g., intramuscularly) in a volume in the range of about 100 μΐ to about 10 ml, containing concentrations of about 10 to 10 virus particles. / ml. Preferably, the human adenoviral vector is administered in a volume in the range of 0.1 to 2.0 ml. For example, the human adenoviral vector can be administered in a volume of 100 μΐ, 500 μΐ, 1 ml, 2 ml. More preferably, the human adenoviral vector is administered in a volume of 0.5 ml. Optionally, the human adenoviral vector can be administered at a concentration of about 10 pv / ml, 10 pv / ml, 10 pv / ml, 10 pv / ml, 5 x 1010 pv / ml, 1011 pv / ml or 1012 pv / ml. (0101) Normally, the human adenoviral vector is administered in an amount of from about 10 to about 10 virus particles (pv) to a human subject during administration, more usually in an amount of about 1010 to about 1012 pv . The initial vaccination is followed by a booster as indicated above. (0102) In another example scheme, the simian adenoviral (e.g., chimpanzee) vector is administered (e.g., intramuscularly) together with a pharmaceutically acceptable carrier. The simian adenoviral vector (e.g. chimpanzee) can be administered, for example, with saline, in the range of about 100 μl to about 10 ml of saline containing concentrations of about 10 to 10 virus particles / ml . For example, the simian adenoviral vector (for example chimpanzee) can be administered with 100 μΐ, 500 μΐ, 1 ml, 2 ml, 3 ml, 4 ml, 5 ml, 6 ml, 7 ml, 8 ml, 9 ml or 10 ml. ml of saline solution. Optionally the simian adenoviral vector (e.g. chimpanzee) can be administered at a concentration of about 104 pv / ml, 105 pv / ml, 106 pv / ml, 107 pv / ml, OQ 1 Γ) 11 ΙΟ 10 pv / ml ml, 10 μv / ml, 10 μv / ml, 10 μv / ml or 10 μv / ml. Typically, the simian adenoviral (e.g., chimpanzee) vector is administered to a human subject during administration in an amount of from about 10 9 to about 10 virus particles (pv), more typically, from about 10 to about 10 pv. (0103) The initial vaccination is followed by a booster, as indicated above; the composition may, if desired, be present in a kit, package or dispenser, which may contain one or more unit dosage forms containing the active component. The kit, for example, may comprise a sheet of metal or plastic, such as a thermoformed package. The kit, packaging or dispenser may be accompanied by instructions for administration. (0104) The compositions of the invention may be administered alone or in combination with other treatments, either simultaneously or successively, depending on the condition to be treated. (0105) The invention also provides the following non-limiting embodiments: (0106) 1. A vaccine combination comprising: (i) a first composition comprising an immunologically effective amount of at least one human adenoviral vector comprising a first acid nucleic acid encoding a first antigenic protein of a filovirus, and a pharmaceutically acceptable carrier; and (ii) a second composition comprising an immunologically effective amount of a simian adenoviral vector comprising a second nucleic acid encoding a second filovirus antigenic protein, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (0107) 2. The combination of vaccine according to embodiment 1, wherein the first antigenic protein comprises an amino acid sequence having a sequence identity of at least 90% with the amino acid sequence of SEQ ID No. 1, preferably is also capable of inducing an immune response to the amino acid sequence of SEQ ID No. 1. (0108) 3. The combination of vaccine according to embodiment 1 or 2, wherein the first antigenic protein comprises an amino acid sequence having a sequence identity of at least 95% with the amino acid sequence of SEQ ID No. 1, preferably is also capable of inducing an immune response to the amino acid of SEQ ID No. 1. (0109) 4. The combination of vaccine according to any one of embodiments 1 to 3, wherein the first antigenic protein comprises the amino acid sequence of SEQ ID No. 1. (0110) 5 The combination of vaccine according to any one of embodiments 1 to 3, wherein the first antigenic protein comprises the amino acid sequence of SEQ ID No. 3. (0111) 6. The combination of vaccine according to any of embodiments 1 to 5, wherein the second antigenic protein comprises an amino acid sequence having a sequence identity of at least 90% with the amino acid sequence of SEQ ID NO: 1, preferably is also capable to induce an immune response to the amino acid sequence of SEQ ID No. 1. (0112) 7. The vaccine combination of any one of embodiments 1 to 6, wherein the second antigenic protein comprises a amino acid sequence having a sequence identity of at least 95% with the amino acid sequence of SEQ ID No. 1, preferably is also capable of inducing an immune response to the amino acid sequence The combination of vaccine according to any one of embodiments 1 to 7, wherein the second antigenic protein comprises the amino acid sequence of SEQ ID No. 1. (0114) 9. The combination of vaccine according to any one of embodiments 1 to 7, wherein the second antigenic protein comprises the amino acid sequence of SEQ ID No. 3. (0115) 10. The combination of vaccine according to any one of embodiments 1 to 9, wherein the human adenoviral vector is a rAd26 vector. (0116) 11. The combination of vaccine according to any one of embodiments 1 to 10, wherein the simian adenoviral vector is an adenoviral chimpanzee vector. (0117) 12. The combination of vaccine according to embodiment 11, wherein the chimpanzee adenoviral vector is a ChAd3 vector. (0118) 13. A vaccine combination comprising: a. a first composition comprising an immunologically effective amount of a rAd26 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ K) No. 1, and a pharmaceutically acceptable carrier; and B. a second composition comprising an immunologically effective amount of a ChAd3 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (0119) 14. A vaccine combination comprising: a. a first composition comprising an immunologically effective amount of a rAd26 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID NO: 3, and a pharmaceutically acceptable carrier; and B. a second composition comprising an immunologically effective amount of a ChAd3 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (0120) 15. A vaccine combination comprising: a. a first composition comprising an immunologically effective amount of a rAd26 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID NO: 1, and a pharmaceutically acceptable carrier; and B. a second composition comprising an immunologically effective amount of a ChAd3 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID NO: 3, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (0121) 16. The combination of vaccine according to any one of embodiments 1 to 15 for use in generating a protective immune response in a subject in need of such response against at least one subtype filovirus, the first composition being used for the priming of said immune response and the second composition being used for the booster of said immune response. (0122) 17. The combination of vaccine according to any one of embodiments 1 to 15 for use in generating a protective immune response in a subject in need of such response against at least one subtype filovirus, the second composition being used for priming said immune response and the first composition being used for boosting said immune response. (0123) 18. A method of inducing an immune response against a filovirus in a subject in need of such response, the method comprising: a. administering to a first composition comprising an immunologically effective amount of a human adenoviral vector comprising a first nucleic acid encoding a first filovirus antigenic protein; and B. administering to a second composition comprising an immunologically effective amount of a simian adenoviral vector comprising a second nucleic acid encoding a second filovirus antigenic protein, wherein steps (a) and (b) are performed in that order or in reverse order. (0124) 19. The method according to embodiment 18, wherein the first antigenic protein comprises an amino acid sequence having a sequence identity of at least 90% with the amino acid sequence of SEQ ID No. 01, preferably is also capable of inducing an immune response to the amino acid sequence of SEQ ID No. 1. (0125) 20. The method according to embodiment 18 or 19, wherein the first antigenic protein comprises a sequence of amino acids having a sequence identity of at least 95% with the amino acid sequence of SEQ ID No. 1, preferably is also capable of inducing an immune response to the amino acid sequence of SEQ ID No. 1. (0126) 21. The method of any one of embodiments 18 to 20, wherein the first antigenic protein comprises the amino acid sequence of SEQ ID No. 1. (0127) 22. The method according to any one of any of Embodiments 18 to 20, wherein the first antigenic protein comprises the amino acid sequence of SEQ ID No. 3. (0128) 23. The method according to any one of embodiments 18 to 22, wherein second antigenic protein comprises an amino acid sequence having a sequence identity of at least 90% with the amino acid sequence of SEQ ID No. 1, preferably is also capable of inducing an immune response to the amino acid of SEQ ID No. 1. (0129) 24. The method according to any one of embodiments 18 to 23, wherein the second antigenic protein comprises an amino acid sequence having a sequence identity of at least 95% with the amino acid sequence of SEQ ID No. 1, preferably is also capable of inducing an immune response to the amino acid sequence of SEQ ID No. 1. (0130) 25. The process according to any of the Embodiments 18 to 24, wherein the second antigenic protein comprises the amino acid sequence of SEQ ID No. 1. (0131) 26. The method according to any one of embodiments 18 to 24, wherein the second antigenic protein comprises the amino acid sequence of SEQ ID No. 3. (0132) 27. The method of any one of embodiments 18 to 26, wherein the human adenoviral vector is a rAd26 vector. (0133) 28. The method according to any one of embodiments 18 to 27, wherein the simian adenoviral vector is a ChAd3 vector. (0134) 29. A method of inducing an immune response against a filovirus in a subject in need of such response, the method comprising: a. administering to a first composition comprising an immunologically effective amount of a rAd26 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID No. 1, and a pharmaceutically acceptable carrier; and B. administering to a second composition comprising an immunologically effective amount of a ChAd3 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID No. 1 or SEQ ID No. 3, and a pharmaceutically acceptable vehicle, steps (a) and (b) being performed in this order or in the reverse order. (0135) 30. A method of inducing an immune response against a filovirus in a subject in need of such response, the method comprising: a. administering to a first composition comprising an immunologically effective amount of a rAd26 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID No. 3, and a pharmaceutically acceptable carrier; and B. administering to a second composition comprising an immunologically effective amount of a ChAd3 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID No. 1 or SEQ ID No. 3, and a pharmaceutically acceptable vehicle, steps (a) and (b) being performed in this order or in the reverse order. (0136) 31. A method of inducing an immune response against a filovirus in a subject in need of such response, the method comprising: a. administering to a first composition comprising an immunologically effective amount of a rAd26 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID No. 1, and a pharmaceutically acceptable carrier; and B. administering to a second composition comprising an immunologically effective amount of a ChAd3 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID NO: 3, and a pharmaceutically acceptable carrier, the steps (a) and (b) being performed in this order or in reverse order. (0137) 32. The method according to any one of embodiments 18 to 31, wherein step (b) is carried out 1-15 weeks, preferably 1-12 weeks, after step (a). (0138) 33. The method according to any one of embodiments 18 to 31, wherein step (b) is carried out 1-15 weeks, preferably 1-12 weeks, before step (a). (0139) 34. A kit comprising: a. a first composition comprising an immunologically effective amount of at least one human adenoviral vector comprising a first nucleic acid encoding a first antigenic protein of a filovirus, and a pharmaceutically acceptable carrier; and B. a second composition comprising an immunologically effective amount of a simian adenoviral vector comprising a second nucleic acid encoding a second antigenic protein, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (0140) 35. The kit according to embodiment 34, wherein the first antigenic protein comprises an amino acid sequence having a sequence identity of at least 90% with the amino acid sequence of SEQ ID No. 1, Preferably, it is also capable of inducing an immune response to the amino acid sequence of SEQ ID No. 1. (0141) 36. The kit according to embodiment 34 or 35, wherein the first antigenic protein comprises a sequence of amino acids having a sequence identity of at least 95% with the amino acid sequence of SEQ ID No. 1, preferably is also capable of inducing an immune response to the amino acid sequence of SEQ ID No. 1. (0142) 37. The kit according to any one of embodiments 34 to 36, wherein the first antigenic protein comprises the amino acid sequence of SEQ ID No. 1. (0143) 38. The kit according to one qu each of embodiments 34-36, wherein the first antigenic protein comprises the amino acid sequence of SEQ ID NO: 3 (0144) 39. The kit according to any of embodiments 34-38, in which wherein the second antigenic protein comprises an amino acid sequence having a sequence identity of at least 90% with the amino acid sequence of SEQ ID No. 1, preferably is also capable of inducing an immune response to the amino acid sequence of SEQ ID No. 1. (0145) 40. The kit according to any one of embodiments 34 to 39, wherein the second antigenic protein comprises an amino acid sequence having a sequence identity at least 95% with the amino acid sequence of SEQ ID No. 1, preferably is also capable of inducing an immune response to the amino acid sequence of SEQ ID No. 1. (0146) 41. The necessary according to the any one of embodiments 34 to 40, wherein the second antigenic protein comprises the amino acid sequence of SEQ ID No. 1. (0147) 42. The kit according to any one of embodiments 34 to 40, wherein the second antigenic protein comprises the amino acid sequence of SEQ ID NO: 3 (0148) 43. The kit according to any one of embodiments 34 to 42, wherein the human adenoviral vector is a rAd26 vector . (0149) 44. The kit according to any one of embodiments 34 to 43, wherein the simian adenoviral vector is a ChAd3 vector. (0150) 45. A kit comprising: a. a first composition comprising an immunologically effective amount of a rAd26 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID NO: 1, and a pharmaceutically acceptable carrier; and B. a second composition comprising an immunologically effective amount of a ChAd3 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (0151) 46. A kit comprising: a. a first composition comprising an immunologically effective amount of a rAd26 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID NO: 3, and a pharmaceutically acceptable carrier; and B. a second composition comprising an immunologically effective amount of a ChAd3 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (0152) 47. A kit comprising: a. a first composition comprising an immunologically effective amount of a rAd26 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID No. 1, and a pharmaceutically acceptable carrier; and B. a second composition comprising an immunologically effective amount of a ChAd3 vector comprising a nucleic acid encoding an antigenic protein comprising the amino acid sequence of SEQ ID NO: 3, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition. (0153) 48. The kit according to any one of embodiments 34 to 47, for use in generating a protective immune response against at least one filovirus subtype, wherein the first composition is used for priming said immune response and the second composition is used for boosting said immune response. (0154) 49. The kit according to any one of embodiments 34 to 47, for use in generating a protective immune response against at least one filovirus subtype, wherein the second composition is used for priming said immune response and the first composition is used for boosting said immune response. (0155) 50. The kit according to any one of embodiments 34 to 49, wherein the priming composition is administered to a subject in need 1-15 weeks, preferably 1-12 weeks, prior to administering the recall composition to the subject.
EXAMPLES (0156) The following examples are given by way of illustration, but without limitation, of the claimed invention.
Phase I Trial in Humans of a ChAd3-EBO-Z and Ad26.ZEBOV Combination Primer-Booster Scheme (0157) A Phase I clinical study in humans was designed to assess the safety of the heterologous Recall combining the monovalent candidate vaccines against Ebola Zaire ChAd3-EBO-Z and Ad26.ZEBOV at intervals of 4 or 8 weeks. The experimental sample consisted of 32 healthy adult subjects aged 18 to 50 years. (0158) ChAd3-EBO-Z is a viral vectorized vaccine using a chimpanzee adenovirus as a vector encoding a Zaire strain Ebola glycoprotein. It was administered at a dose of 1 x 10 pv. Ad26.ZEBOV is a viral vectorized vaccine using a human adenovirus as vector encoding a glycoprotein of Ebola strain Zaire virus. . It was administered at a dose of 5 x 1010 pv. ChAd3-EBO-Z and Ad26.ZEBOV were each administered intramuscularly according to the experimental setup shown in Table 1.
Table 1: Experimental setup for the Phase I trial in human subjects of a primer-booster regimen combining ChAd3-EBO-Z and Ad26.ZEBOV
(0159) Group 1 (Gl) candidates received a ChAd3-EBO-Z priming vaccine at week 0 and an Ad26.ZEBOV booster vaccine at week 4. Group 2 (G2) candidates ) received an Ad26.ZEBOV priming vaccine at week 0 and a ChAd3-EBO-Z booster vaccine at week 4. Group 3 (G3) candidates received a ChAd3 priming vaccine -EBO-Z at week 0 and a booster vaccine of Ad26.ZEBOV at week 8. Group 4 (G4) candidates received an Ad26.ZEBOV priming vaccine at week 0 and a vaccine ChAd3-EBO-Z at week 8. The follow-up duration of the study was 5 months.
Vaccine materials (0160) The use of the recombinant replication-incompetent ChAd3 vector for the construction of the ChAd3-EBO-2 vaccine was based on the as easy induction properties of immune responses as rAd5, but with immunity pre-existing low or none against ChAd3 in human populations. The recombinant Ebola Zaire vaccine vectorized with adenovirus type 3 chimpanzee (ChAd3-EBO-Z) is a replication-deficient recombinant serotype 3 adenovirus vector (ChAd3), expressing a glycoprotein having the amino acid sequence of SEQ ID No. 3, which is identical to the wild-type (WT) Ebola glycoprotein (SEQ ID No. 1) derived from the Zaire strain, except for the presence of a valine at position 662 instead of an isoleucine. The ChAd3-EBO-Z vaccine has been developed using the adenovirus vaccine platform technology. The drug substance has been manufactured and labeled under Good Manufacturing Practice (GMP) conditions. ChAd3-EBO-Z was provided as a sterile aliquot liquid in 1 ml clear glass vials, at a concentration of 11 11 1 x 10 pv per ml and in a 2 ml aliquot at a concentration of 1 ml. concentration of 1 x 10 pv per ml. (0161) The human adenovirus Ad26.ZEBOV, which encodes Ebola WT GP (SEQ ID No. 1) was provided as a liquid in sterile aliquots at a concentration of 1 x 1010 pv per ml. The vaccines were stored in a freezer at -80 ° C.
Vaccination and Experimental Design (0162) The vaccines were administered intramuscularly (IM) to all groups in the deltoid muscle of either arm. On the day of vaccination, vaccines were allowed to thaw at room temperature and administered within one hour. Depending on the dose and concentration, in general, one or more vials of vaccine may be used. Likewise, in general, one vial can be used for more than one vaccine if the administration is carried out within the limits of the post-defrosting time period, ie 1 hour. (0163) On vaccination days, subjects in the study underwent clinical evaluation prior to injection and samples were taken for laboratory tests. (0164) The specific immunogenicity of Ebolavirus can be evaluated by various immunological assays. In general, the measured results of the primary immunogenicity are ELISA assays and antigen-specific neutralization assays for antibody responses and intracellular cytokine staining assays (ICC) for T-cell responses. Immunogenicity can be assessed using the immunoassays summarized in Tables 3 and 4. The set of exploratory assays may include, but are not limited to, the indicated dosages. Measurements of the exploratory results include, but are not limited to, ex vivo ELISPOT and flow cytometry performed with research samples collected at certain times during the entire study, as well as by other immunogenicity assays and the evaluation of genetic factors associated with immune responses. Vaccine-induced mRNA expression patterns, before and after vaccination, can also be established as an exploratory assessment. (0165) The specific humoral immune response of Ebola glycoproteins was evaluated by ELISA.
Table 3: Summary of immunological (serological) assays
EBOV: Ebola virus; ELISA assay by bound enzyme immunoadsorption;
GP: glycoprotein; IgG: immunoglobulin G
Table 4: Summary of immunological (cellular) assays
EBOV: Ebola virus; ELISpot: bound enzyme immunospot assay; CCI: Intracellular cytokine staining: IFN: interferon: IL: interleukin; PBMCs: mononuclear cells of peripheral blood; TNF: tumor necrosis factor.
ELISA Assay for the Ebola Glycoprotein Specific IgG Response (0166) Briefly, plates were coated with a recombinant, soluble, affinity purified trimer form of the Makona Ebola variant glycoprotein. Antibody responses were measured against the trimerized glycoprotein of Ebola strain Zaire. Samples were assayed in triplicate and a ZEBOV-GP-positive serum reference pool was used to construct a calibration curve on each plate. Arbitrary ELISA units were calculated for each sample using the OD values of the sample and the parameters of the calibration curve. A threshold of seropositivity of 166 ELISA units was determined from the average of ELISA + 3 units and 59 naive samples tested in the assay. (0167) Humoral immune responses were measured for all volunteers up to 28 days after booster vaccination (B + 28) and for 17 of 32 volunteers up to 90 days after booster vaccination ( B + 90) (G1: n = 7. G2: n = 5. G3: n = 3, G4: n = 2). The remaining B + 90 samples were analyzed after completion of all visits. (0168) FIGs. 1 and 2 are graphs showing the kinetics of the GP EBOV specific humoral immune response. evaluated by ELISA. FIG. 1 summarizes data obtained from test groups G1 and G3. FIG. 2 summarizes the data obtained from test groups G2 and G4. The geometric mean ELISA concentrations are expressed in ELISA units per ml, together with their 95% confidence interval. Group 1 (Gl) candidates received primer immunization with ChAd3 expressing GP EBOV, followed by a booster with Ad26 expressing GP EBOV, 4 weeks later. Group 2 (G2) candidates received a primer immunization with Ad26, followed by a booster with ChAd3, 4 weeks later. Group 3 (G3) candidates received primer immunization with ChAd3 followed by a booster with Ad26 8 weeks later. Group 4 (G4) candidates received primer immunization with Ad26 followed by booster with ChAd3 8 weeks later. (0169) As shown in the graphs of FIGs. 1 and 2, immune responses were induced by both adenoviral vectors from chimpanzees and adenoviral vectors of human origin, expressing Ebola Zaire GP. It is important to note that responses increased significantly after booster with the heterologous vector. At the peak of the immune response observed 2 weeks after the booster, GP EBOV-specific antibody titers increased 3.7- to 7.6-fold compared to pre-booster when booster immunization was administered 4 weeks later. immunization primer (Gl and G2) and 8.6 to 12.4 fold when administered 8 weeks after (G3 and G4). Overall, a slightly higher immune response was observed when a longer interval between primer and booster was used. It is interesting to note that at 90 days post-booster immunization (the last moment analyzed), GP EBOV-specific humoral immune responses were maintained at a rate exceeding the response observed after primer immunization. (0170) In conclusion, heterologous primer-boosting regimens using an adenoviral vaccine vector derived from a chimpanzee and an adenoviral vaccine vector of human origin are capable of inducing a strong and long-lasting immune response, regardless of the order of administration of the vaccines as priming or booster immunization.
SEQUENCE LISTING <110> Crucell Holland B.V.
Glaxosmithkline Biologicals S.A.
VAN HOOF, Johan Jules Urbain SLAOUI, Moncef BALLOU, Ripley W <120> Methods and Compositions for Inducing Protective Immunity Against
Filovirus Infection and / or Disease <130> 688097-70BE <160> 3 <170> PatentIn version 3.5 <210> 1 <211> 676
<212> PRT <213> Artificial Sequence <220> <223> Wild-type (WT) Ebola Glycoprotein (GP) <400> 1
Met Gly Val Thr Gly Island Leu Gin Leu Pro Arg Asp Arg Phe Lys Arg 1 5 10 15
Thr Ser Phe Phe Leu Trp Val Ile Ile Leu Phe Gin Arg Thr Phe Ser 20 25 30
Ile Pro Leu Gly Val Ile His Asn Ser Thr Leu Gin Val Ser Asp Val 35 40 45
Asp Lys Leu Val Cys Arg Asp Lys Leu Ser Ser Thr Asn Gin Leu Arg 50 55 60
Ser Val Gly Leu Asn Leu Glu Gly Asn Gly Ala Thr Val Asp Val Pro 65 70 75 80
Ser Ala Thr Lys Arg Trp Gly Phe Arg Ser Pro Gly Val Pro Lys Val 85 90 95
Val Asn Tire Glu Ala Gly Glu Trp Ala Glu Asn Cys Tire Asn Leu Glu 100 105 110
Ile Lys Lys Pro Asp Gly Glu Cys Leu Pro Ala Ala Pro Asp Gly 115 120 125
Isle Arg Gly Phe Pro Arg Cys Arg Tyr Val His Lys Val Ser Gly Thr 130 135 140
Gly Pro Cys Ala Gly Asp Phe Ala Phe His Lys Glu Gly Ala Phe Phe 145 150 155 igo
Leu Tyr Asp Arg Arg Leu Ala Ser Thr Val Isle Tyr Arg Gly Thr Thr Phe 165 170 175
Ala Glu Gly Val Val Ala Phe Leu Leu Leu Pro Gin Ala Lily Lys Asp 180 185 190
Phe Phe Ser Ser His Pro Leu Arg Glu Pro Asn Val Ala Thr Glu Asp 195 200 205
Pro Ser Ser Gly Tire Tyr Ser Thr Thr Isle Arg Tyr Gin Ala Thr Gly 210 215 220
Phe Gly Thr Asn Glu Thr Glu Tyr Leu Phe Asp Glu Asp Asn Leu Thr 225 230 235 240
Tyr Val Gin Leu Glu Ser Arg Phe Thr Pro Gin Phe Leu Leu Gin Leu 245 250 255
Asn Glu Thr Tyr Tyr Ser Ser Gly Lys Arg Ser Asn Thr Thr Gly Lys 260 265 270
Leu Ile Trp Lys Val Asn Pro Glu Asp Asp Thr Thr Ile Gly Glu Trp 275 280 285
Ala Phe Trp Glu Thr Lys Lily Asn Leu Thr Arg Lys Island Arg Ser Glu 290 295 300
Glu Leu Ser Phe Thr Val Ser Val Asn Gly Ala Lys Asn Ser Gly Island 305 310 315 320
Gin Ser Pro Ala Arg Ser Ser Asp Asp Pro Gly Thr Thr Thr Thr 325 330 335
Glu Asp His Lys Met Ala Ser Glu Asn Ser Ser Ala Met Val Gin 340 345 350
Val His Ser Gin Gly Arg Ala Gla Ala Val Ser His Leu Thr Leu Leu 355 360 365
Ala Thr Ser Ser Ser Pro Ser Pro Gin Ser Pro Thr Thr Lys Pro Gly Pro 370 375 380
Asp Asn Ser Thr His Asn Thr Pro Val Tyr Lys Leu Asp Asp Ser Ser Glu 385 390 395 400
Ala Thr Gin Val Glu Gin His His Arg Arg Asp Asp Asn Asp Ser Thr 405 410 415
Ala Ser Asp Thr Pro Ser Ala Thr Thr Ala Pro Ala Gly Pro Lys Ala 420 425 430
Glu Asn Thrn Asn Thr Ser Lys Ser Thr Asp Asp Phe Leu Asp Pro Ala Thr 435 440 445
Thr Thr Ser Pro Gin Asn His Ser Thr Glu Ala Gly Asn Asn Asn Thr 450 455 460
His His Gin Asp Thr Gly Glu Ser Ser Ser Ser Gly Gly Lys Leu Gly 465 470 475 480
Leu Thr Island Thr Thrn Ala Gly Val Ala Gly Leu Thr Thr Gly Gly 485 490 495
Arg Arg Thr Arg Arg Glu Ala Val Asn Ala Gin Pro Lys Cys Asn 500 505 510
Pro Asn Leu His Tyr Thr Trp Thr Asp Asp Glin Glly Ala Ala Gly Island 515 520 525
Leu Ala Trp Pro Island Tyr Phe Gly Pro Ala Ala Glu Gly Island Tire Island 530 535 540
Glu Gly Leu Met His Asn Gin Asp Gly Leu Ile Cys Gly Leu Arg Gin 545 550 555 560
Leu Ala Asn Thr Glu Thr Gin Ala Leu Gin Leu Phe Leu Arg Ala Thr 565 570 575
Thr Glu Leu Arg Thr Phe Ser Ile Leu Asn Arg Lys Ala Ile Asp Phe 580 585 590
Leu Leu Gin Arg Trp Gly Gly Thr Cys His Leu Gly Pro Island Asp Cys 595 600 605
Cys Island Glu Pro His Asp Trp Thr Lys Asn Island Thr Asp Lys Island Asp 610 615 620
Gin Ile Ile His Asp Phe Asp Val Asp Lys Thr Pro Leu Asp Gin Gly Asp 625 630 635 640
Asn Asp Asn Trp Thr Trp Gly Trp Arg Gin Trp Pro Island Ala Gly Island 645 650 655
Gly Val Thr Gly Val Ile Ile Ala Val Ile Ala Leu Phe Cys Ile Cys 660 665 670
Lys Phe Val Phe 675
<210> 2 <211> 2028 <212> DNA <213> Artificial Sequence <220> <223> DNA wild-type (WT) Ebola glycoprotein (GP) <400> 2 atgggcgtga ccggcatcct gcagctgccc agagaccggt tcaagcggac cagcttcttc 60 ctgtgggtga tcatcctgtt ccagcggacc ttcagcatcc ccctgggcgt gatccacaac 120 agcaccctgc aggtgtccga cgtggacaag ctggtgtgcc gggacaagct gagcagcacc 180 aaccagctgc ggagcgtggg cctgaacctg gaaggcaatg gcgtggccac cgacgtgccc 240 agcgccacaa agagatgggg cttcagatcc ggcgtgcccc ccaaggtggt gaactatgag 300 gccggcgagt gggccgagaa ctgctacaac ctggaaatca agaagcccga cggcagcgag 360 tgcctgcctg ccgctcctga tggcatcaga ggcttccccc ggtgcagata cgtgcacaag 420 gtgtccggca ccggcccctg tgccggcgat ttcgccttcc acaaagaggg cgcctttttc 480 ctgtacgacc ggctggccag caccgtgatc taccggggca ccaccttcgc cgaaggcgtg 540 gtggccttcc tgatcctgcc ccaggccaag aaggacttct tcagcagcca ccccctgcgc 600 gagcccgtga atgccaccga ggatcccagc agcggctact acagcaccac catcagatac 660 caggccaccg gcttcggcac caacgagaca gagtacctgt tcgaggtgga caacctgacc 720 tacgtgcagc tggaaagccg gttcaccccc cagtttctgc tgcagctgaa cgagacaatc 780 tacaccagcg gcaagcggag caacaccaca ggcaagctga tctggaaagt gaaccccgag 840 atcgacacca caatcggaga gtgggccttc tgggagacaa agaagaacct gacccggaag 900 atcagaagcg aggaactgag cttcaccgtg gtgtccaacg gcgccaagaa catcagcggc 960 cagagccctg ccagaacaag cagcgacccc ggcaccaaca ccaccaccga ggaccacaag 1020 atcatggcca gcgagaacag cagcgccatg gtgcaggtgc acagccaggg cagagaagcc 1080 gccgtgtccc acctgaccac cctggccacc atcagcacca gcccccagag cctgaccacc 1140 aagcctggcc ccgacaactc cacccacaac acccccgtgt acaagctgga catcagcgag 1200 gccacccagg tggaacagca ccacagacgg accgacaacg acagcaccgc cagcgatacc 1260 ccctccgcca caacagctgc cggacctccc aaggccgaga acaccaacac ctccaagagc 1320 accgactttc tggaccccgc caccaccaca agcccccaga accactccga gacagccggc 1380 aacaacaaca cccaccatca ggacaccggc gaggaaagcg ccagctctgg caagctgggc 1440 ctgatcacca acacaatcgc cggcgtggcc ggactgatca ccggcggcag aagaaccaga 1500 cgcgaggcca tcgtgaacgc ccagcccaag tgcaacccca acctgcacta ctggaccacc 1560 caggacgagg gcgctgccat tggcctggcc tggatccctt acttcggcc c tgccgccgag 1620 ggcatctaca tcgagggcct gatgcacaac caggacggcc tgatctgcgg cctgcggcag 1680 ctggccaacg aaaccacaca ggccctgcag ctgttcctgc gggccaccac agagctgcgg 1740 accttctcca tcctgaacag aaaggccatc gactttctgc tgcagcgctg gggcggcaca 1800 tgtcacatcc tgggccccga ctgctgcatc gagccccacg actggaccaa gaatatcacc 1860 gacaagatcg accagatcat ccacgacttc gtggacaaga ccctgcccga ccagggcgac 1920 aacgataact ggtggacagg ctggagacag tggatccctg ccggcattgg cgtgacaggc 1980 gtgatcattg ccgtgatcgc cctgttctgc atctgcaagt tcgtgttc 2028 <210> 3 <211> 676
<212> PRT <213> Artificial Sequence <220> <223> Ebola Glycoprotein (GP) <400> 3
Met Gly Val Thr Gly Island Leu Gin Leu Pro Arg Asp Arg Phe Lys Arg 1 5 10 15
Thr Ser Phe Phe Leu Trp Val Ile Ile Leu Phe Gin Arg Thr Phe Ser 20 25 30
Ile Pro Leu Gly Val Ile His Asn Ser Thr Leu Gin Val Ser Asp Val 35 40 45
Asp Lys Leu Val Cys Arg Asp Lys Leu Ser Ser Thr Asn Gin Leu Arg 50 55 60
Ser Val Gly Leu Asn Leu Glu Gly Asn Gly Ala Thr Val Asp Val Pro 65 70 75 80
Ser Ala Thr Lys Arg Trp Gly Phe Arg Ser Pro Gly Val Pro Lys Val 85 90 95
Val Asn Tire Glu Ala Gly Glu Trp Ala Glu Asn Cys Tire Asn Leu Glu 100 105 no
Ile Lys Lys Pro Asp Gly Glu Cys Leu Pro Ala Ala Pro Asp Gly 115 120 125
Isle Arg Gly Phe Pro Arg Cys Arg Tyr Val His Lys Val Ser Gly Thr 130 135 140
Gly Pro Cys Ala Gly Asp Phe Ala Phe His Lys Glu Gly Ala Phe Phe 145 150 155 160
Leu Tyr Asp Arg Arg Leu Ala Ser Thr Val Isle Tyr Arg Gly Thr Thr Phe 165 170 175
Ala Glu Gly Val Val Ala Phe Leu Leu Leu Pro Gin Ala Lily Lys Asp 180 185 190
Phe Phe Ser Ser His Pro Leu Arg Glu Pro Asn Val Ala Thr Glu Asp 195 200 205
Pro Ser Ser Gly Tire Tyr Ser Thr Thr Isle Arg Tyr Gin Ala Thr Gly 210 215 220
Phe Gly Thr Asn Glu Thr Glu Tyr Leu Phe Asp Glu Asp Asn Leu Thr 225 230 235 240
Tyr Val Gin Leu Glu Ser Arg Phe Thr Pro Gin Phe Leu Leu Gin Leu 245 250 255
Asn Glu Thr Tyr Tyr Ser Ser Gly Lys Arg Ser Asn Thr Thr Gly Lys 260 265 270
Leu Ile Trp Lys Val Asn Pro Glu Asp Asp Thr Thr Ile Gly Glu Trp 275 280 285
Ala Phe Trp Glu Thr Lys Lily Asn Leu Thr Arg Lys Island Arg Ser Glu 290 295 300
Glu Leu Ser Phe Thr Val Ser Val Asn Gly Ala Lys Asn Ser Gly Island 305 310 315 320
Gin Ser Pro Ala Arg Ser Ser Asp Asp Pro Gly Thr Thr Thr Thr 325 330 335
Glu Asp His Lys Met Ala Ser Glu Asn Ser Ser Ala Met Val Gin 340 345 350
Val His Ser Gin Gly Arg Ala Gla Ala Val Ser His Leu Thr Leu Leu 355 360 365
Ala Thr Ser Ser Ser Pro Ser Pro Gin Ser Pro Thr Thr Lys Pro Gly Pro 370 375 380
Asp Asn Ser Thr His Asn Thr Pro Val Tyr Lys Leu Asp Asp Ser Ser Glu 385 390 395 400
Ala Thr Gin Val Glu Gin His His Arg Arg Asp Asp Asn Asp Ser Thr 405 410 415
Ala Ser Asp Thr Pro Ser Ala Thr Thr Ala Pro Ala Gly Pro Lys Ala 420 425 430
Glu Asn Thrn Asn Thr Ser Lys Ser Thr Asp Asp Phe Leu Asp Pro Ala Thr 435 440 445
Thr Thr Ser Pro Gin Asn His Ser Thr Glu Ala Gly Asn Asn Asn Thr 450 455 460
His His Gin Asp Thr Gly Glu Ser Ser Ser Ser Gly Gly Lys Leu Gly 465 470 475 480
Leu Thr Island Thr Thrn Ala Gly Val Ala Gly Leu Thr Thr Gly Gly 485 490 495
Arg Arg Thr Arg Arg Glu Ala Val Asn Ala Gin Pro Lys Cys Asn 500 505 510
Pro Asn Leu His Tyr Thr Trp Thr Asp Asp Glin Glly Ala Ala Gly Island 515 520 525
Leu Ala Trp Pro Island Tyr Phe Gly Pro Ala Ala Glu Gly Island Tire Island 530 535 540
Glu Gly Leu Met His Asn Gin Asp Gly Leu Ile Cys Gly Leu Arg Gin 545 550 555 560
Leu Ala Asn Thr Glu Thr Gin Ala Leu Gin Leu Phe Leu Arg Ala Thr 565 570 575
Thr Glu Leu Arg Thr Phe Ser Ile Leu Asn Arg Lys Ala Ile Asp Phe 580 585 590
Leu Leu Gin Arg Trp Gly Gly Thr Cys His Leu Gly Pro Island Asp Cys 595 600 605
Cys Island Glu Pro His Asp Trp Thr Lys Asn Island Thr Asp Lys Island Asp 610 615 620
Gin Ile Ile His Asp Phe Asp Val Asp Lys Thr Pro Leu Asp Gin Gly Asp 625 630 635 640
Asn Asp Asn Trp Thr Trp Gly Trp Arg Gln Trp Pro Island Ala Gly Island 645 650 655
Gly Val Thr Gly Val Val Ile Ala Val Ile Ala Leu Phe Cys Ile Cys 660 665 670
Lys Phe Val Phe 675

权利要求:
Claims (21)
[1]
A combination of vaccine comprising: a. a first composition comprising an immunologically effective amount of at least one human adenoviral vector comprising a first nucleic acid encoding a first antigenic protein of a filovirus, and a pharmaceutically acceptable carrier; and B. a second composition comprising an immunologically effective amount of a simian adenoviral vector comprising a second nucleic acid encoding a second filovirus antigenic protein, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition.
[2]
The vaccine combination according to claim 1, wherein the first antigenic protein comprises the amino acid sequence of SEQ ID No. 1 or SEQ ID No. 3.
[3]
The vaccine combination of claim 1 or 2, wherein the second antigenic protein comprises the amino acid sequence of SEQ ID NO: 1 or SEQ ID NO: 3.
[4]
The vaccine combination according to any one of claims 1 to 3, wherein the human adenoviral vector is a rAd26 vector.
[5]
A vaccine combination according to any one of claims 1 to 4, wherein the simian adenoviral vector is an adenoviral chimpanzee vector.
[6]
The vaccine combination of claim 5, wherein the chimpanzee adenoviral vector is a ChAd3 vector.
[7]
A vaccine combination according to any one of claims 1 to 6 for use in generating a protective immune response against at least one filovirus subtype, wherein the first composition is used for priming said immune response and the second composition is used for the booster of said immune response.
[8]
A vaccine combination according to any one of claims 1 to 6 for use in generating a protective immune response against at least one filovirus subtype, wherein the second composition is used for priming said immune response and the first composition is used for the booster of said immune response.
[9]
A method of inducing an immune response against a filovirus in a subject, the method comprising: a. administering to a first composition comprising an immunologically effective amount of a human adenoviral vector comprising a first nucleic acid encoding a first filovirus antigenic protein; and B. administering to a second composition comprising an immunologically effective amount of a simian adenoviral vector comprising a second nucleic acid encoding a second filovirus antigenic protein, wherein steps (a) and (b) are performed in that order or in reverse order.
[10]
The method of claim 9, wherein the first antigenic protein comprises the amino acid sequence of SEQ ID No. 1 or SEQ ID No. 3.
[11]
The method of claim 9 or 10, wherein the second antigenic protein comprises the amino acid sequence of SEQ ID No. 1 or SEQ ID No. 3.
[12]
The method of any one of claims 9 to 11, wherein the human adenoviral vector is a rAd26 vector.
[13]
The method of any one of claims 9 to 12, wherein the simian adenoviral vector is a ChAd3 vector.
[14]
The method of any one of claims 9 to 13, wherein step (b) is performed 1-15 weeks after step (a).
[15]
15. Required including: a. a first composition comprising an immunologically effective amount of at least one human adenoviral vector comprising a first nucleic acid encoding a first antigenic protein of a filovirus, and a pharmaceutically acceptable carrier; and B. a second composition comprising an immunologically effective amount of a simian adenoviral vector comprising a second nucleic acid encoding a second antigenic protein, and a pharmaceutically acceptable carrier; one of the compositions being a priming composition and the other composition being a booster composition.
[16]
The kit of claim 15, wherein the first antigenic protein comprises the amino acid sequence of SEQ ID No. 1 or SEQ ID No. 3.
[17]
The kit of claim 15 or 16, wherein the second antigenic protein comprises the amino acid sequence of SEQ ID No. 1 or SEQ ID No. 3.
[18]
The kit of any one of claims 15 to 17, wherein the human adenoviral vector is a rAd26 vector.
[19]
The kit of any one of claims 15 to 18, wherein the simian adenoviral vector is a ChAd3 vector.
[20]
A kit according to any of claims 15 to 19 for use in generating a protective immune response against at least one filovirus subtype, wherein the first composition is used for priming said immune response. and the second composition is used for boosting said immune response.
[21]
21. Kit according to any one of claims 15 to 19, for use in generating a protective immune response against at least one filovirus subtype, wherein the second composition is used for priming said immune response and the first composition is used for boosting said immune response.

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同族专利:

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公开号 | 公开日

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BE1023877A1|2017-08-31|

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WO2016187613A1|2016-11-24|

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法律状态:
2017-11-30| FG| Patent granted|Effective date: 20170901 |

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2019-01-30| MM| Lapsed because of non-payment of the annual fee|Effective date: 20180531 |

优先权:

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申请号 | 申请日 | 专利标题

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US201562164909P| true| 2015-05-21|2015-05-21|

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US62164909|2015-05-21|

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