WO2025090476A1 - Vaccins oraux canins et méthodes d'administration - Google Patents

Vaccins oraux canins et méthodes d'administration Download PDF

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Publication number
WO2025090476A1
WO2025090476A1 PCT/US2024/052376 US2024052376W WO2025090476A1 WO 2025090476 A1 WO2025090476 A1 WO 2025090476A1 US 2024052376 W US2024052376 W US 2024052376W WO 2025090476 A1 WO2025090476 A1 WO 2025090476A1
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oral vaccine
canine
recombinant
vaccine
modified live
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Keith Allen Ameiss
Yugang Luo
Mauro Pires MORAES
Sharon Marie Wappel
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Zoetis Services LLC
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Zoetis Services LLC
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/12—Viral antigens
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
    • C12N15/86—Viral vectors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/525—Virus
    • A61K2039/5254—Virus avirulent or attenuated
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/525—Virus
    • A61K2039/5256—Virus expressing foreign proteins
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
    • A61K2039/53—DNA (RNA) vaccination
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/545—Medicinal preparations containing antigens or antibodies characterised by the dose, timing or administration schedule
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
    • A61K2039/552—Veterinary vaccine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/70—Multivalent vaccine
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
    • C12N2710/00011—Details
    • C12N2710/10011—Adenoviridae
    • C12N2710/10311—Mastadenovirus, e.g. human or simian adenoviruses
    • C12N2710/10341—Use of virus, viral particle or viral elements as a vector
    • C12N2710/10343—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector

Definitions

  • the VANGUARD® line of vaccines is used for the vaccination of healthy dogs 6 weeks of age or older as an aid in preventing canine distemper (CD) caused by canine distemper virus (CDV), infectious canine hepatitis (ICH) caused by canine adenovirus type 1 (CAV-1 ), respiratory disease caused by canine adenovirus type 2 (CAV-2), canine parainfluenza (CPI) caused by canine parainfluenza (CPI) virus, and canine parvoviral enteritis caused by canine parvovirus (CPV) (Mouzin DE, et al., 2004, JAVMA, 224: 55-60).
  • CD is a high morbidity, high mortality viral disease occurring in unvaccinated dog populations worldwide. Approximately 50% of non-vaccinated, non-immune dogs infected with CDV develop clinical signs and approximately 90% of those dogs die (Swango LJ.1983. Norden News 58:4-10).
  • the respiratory disease caused by CAV-2 in severe cases, may include pneumonia and bronchopneumonia.
  • CAV- 2 vaccine has been shown to cross-protect against ICH caused by CAV-1 (Bass EP, et al., 1980, JAVMA, 177:234-242).
  • the upper respiratory disease caused by CPI virus may be mild or subclinical, with signs becoming more severe if concurrent infection with other respiratory pathogens exists.
  • the enteric disease caused by CPV is characterized by sudden onset of vomiting and diarrhea, often hemorrhagic, and may be accompanied by leukopenia (Appel MJ, et al., 1979, Vet Rec, 105:156-159). [0004] Generally, puppies are vaccinated at about 6 weeks of age with so-called core vaccines, comprising antigens against CDV, CAV-2, and CPV.
  • the disclosure provides a method of protecting an MDA (Maternally Derived Antibodies) -negative canine against canine distemper virus, the method comprising administering a recombinant oral vaccine comprising a modified live canine adenovirus type 2 (CAV-2) vector carrying a Canine Distemper Virus (CDV) antigen to said canine, wherein the recombinant oral vaccine is administered orally in a first dose and orally in a second dose.
  • MDA Magnetically Derived Antibodies
  • CAV-2 modified live canine adenovirus type 2
  • CDV Canine Distemper Virus
  • the disclosure provides a method of annually revaccinating a dog, the method comprising orally administering to said dog a vaccine comprising a recombinant oral vaccine comprising a modified live canine adenovirus type 2 (CAV-2) vector carrying a Canine Distemper Virus (CDV) antigen, wherein said recombinant oral vaccine is administered about a year after a previous annual revaccination or about a year after administering to said dog a first dose of an initial vaccine, wherein further said first dose of the initial vaccine is administered to said dog parenterally, and wherein said initial vaccine comprises a CDV antigen and a CAV-2 antigen.
  • CAV-2 Canine Distemper Virus
  • the CDV antigen in said initial vaccine, is a) a modified live CDV and the CAV-2 antigen is a modified live CAV-2; or b) encoded by a nucleic acid sequence within a modified live canarypox virus genome.
  • the dog in said previous annual revaccination the dog is administered a vaccine wherein the CDV antigen is a) a modified live CDV and the CAV-2 antigen is a modified live CAV-2; or b) encoded by a nucleic acid sequence within a modified live canarypox virus genome.
  • the CDV antigen in the recombinant oral vaccine, is H protein, preferably comprising SEQ ID NO: 1 or a sequence that is 90% or 95% identical to SEQ ID NO: 1.
  • SEQ ID NO: 1 in the recombinant oral vaccine SEQ ID NO: 1 is encoded by a nucleic acid sequence comprising SEQ ID NO: 2.
  • the recombinant oral vaccine further comprises F protein of CDV, preferably comprising SEQ ID NO: 3 or a sequence that is 90% or 95% identical to SEQ ID NO: 3.
  • SEQ ID NO: 3 is encoded by a nucleic acid sequence comprising SEQ ID NO: 3.
  • the nucleic acid sequence encoding the CDV antigen according to any of the embodiments of the recombinant oral vaccine as described above is inserted into E3 region of the modified live CAV-2 vector genome.
  • said recombinant oral vaccine contains at least 10 3 TCID 50 to about 10 8 TCID 50 of said modified live CAV-2 vector carrying the CDV antigen per dose, more preferably about 10 3.5 TCID 50 to about 10 4.5 TCID 50 of said modified live CAV-2 vector carrying the CDV antigen per dose.
  • the modified live canine adenovirus type 2 vector carrying a CDV antigen protects against Canine Distemper Virus, Canine Adenovirus Type 1 infection, and Canine Adenovirus Type 2 infection when administered orally.
  • the canine vaccinated according to the methods of the first aspect of the invention has not been previously vaccinated against Canine Distemper virus and, more preferably, against Canine Adenovirus Type 2.
  • the recombinant oral vaccine further comprises a modified live parvovirus, wherein said recombinant oral vaccine further protects against Canine Parvovirus infection when administered orally.
  • said vaccine comprises about 10 5.0 TCID 50 to about 10 10 TCID 50 of the modified live Canine Parvovirus, more preferably, 10 7.5 TCID 50 to about 10 9 TCID 50 of the modified live Canine Parvovirus.
  • the recombinant oral vaccine according to any embodiments of the method according to the first or second aspect of the invention further comprises modified live Canine Parainfluenza virus, wherein said recombinant oral vaccine further protects against Canine Parainfluenza virus infection when administered orally.
  • said canine has not been previously vaccinated against Canine Parainfluenza virus.
  • the recombinant oral vaccine comprises 10 4.5 TCID 50 to about 10 8 TCID 50 of the modified live Canine Parainfluenza virus, preferably 10 6.5 TCID 50 to about 10 7.5 TCID 50 of the modified live Canine Parainfluenza virus.
  • said canine has not been previously vaccinated against Canine Parainfluenza virus.
  • said canine is about 8 to about 16 weeks old at the time of the first administration of the recombinant oral vaccine.
  • the canine is at least 13 months old at the time of administering the recombinant oral vaccine.
  • the second dose is administered 7-35 days after the first dose, preferably about 21 days after the first dose.
  • the methods disclosed herein further comprise the step of orally administering a third dose of the recombinant oral vaccine described herein, wherein said third dose is administered about 21 days after the second dose.
  • the recombinant oral vaccine is provided, for use according to the embodiments of the first and the second aspect, respectively, as described above.
  • the following six groups each contain amino acids that are typical conservative substitutions for one another: [1] Alanine (A), Serine (S), Threonine (T); [2] Aspartic acid (D), Glutamic acid (E); [3] Asparagine (N), Glutamine (Q); [4] Arginine (R), Lysine (K), Histidine (H); [5] Isoleucine (I), Leucine (L), Methionine (M), Valine (V); and [6] Phenylalanine (F), Tyrosine (Y), Tryptophan (W), (see, e.g., US Patent Publication 20100291549).
  • references to animals that “have not been vaccinated” or “have not been actively vaccinated” or the like do not encompass maternal immunity or passive antibody transfer.
  • the term “orally” or “oral” or the like refers to administration into the mouth of a subject, where the vaccine contacts the subject’s oral mucosa.
  • the terms “protect”, “protection”, “protective immune response” and the like generally refer to the ability of the vaccines disclosed herein to reduce or eliminate the duration or the severity of at least one clinical sign of the pathogen against which the subjects are vaccinated. The clinical signs vary depending on the pathogen. A more detailed description of the protective immune response to specific viruses is provided below.
  • the term “subject” refers to animals to which the vaccines disclosed herein are administered.
  • the term refers non-human animals, such as dogs, cats, horses, pigs, cattle, swine. In certain embodiments, the term also refers humans.
  • the term "therapeutically effective amount,” in the context of this disclosure, refers to an amount of an antigen or vaccine that would induce a protective immune response in a subject receiving the antigen or vaccine.
  • the term “vaccine” refers to a composition containing an antigen against specific pathogen, wherein the antigen can elicit protective immune response against said pathogen.
  • the response preferably is generated by the adaptive immune system and may include an antibody response generated by B cells, cell-mediated immunity mediated by T-cells, or both antibody- and cell-mediated immunity.
  • the instant disclosure provides a recombinant oral vaccine comprising a modified live canine adenovirus type 2 (CAV-2) vector carrying an antigen from a Canine Distemper Virus.
  • CAV-2 canine adenovirus type 2
  • the antigen from the Canine Distemper virus is its H-protein (Hemagglutinin) or F-protein (Fusion), as these proteins are present on the surface of the virus and are known to elicit protective immune response.
  • the CDV antigen in the recombinant oral vaccine is the H protein.
  • Respective H proteins from different CDV strains are available in publicly accessible databases of genetic information.
  • the H protein comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 1, and can be in different embodiments, at least 91% identical, or at least 92% identical, or at least 93% identical, or at least 94% identical, or at least 95% identical, or at least 96% identical, or at least 97% identical, or at least 98% identical, or at least 99% identical, or 100% identical to SEQ ID NO: 1.
  • H protein is at least 90% identical but less than 100% identical to SEQ ID NO: 1
  • at least half of the differing amino acids are conservative substitutions.
  • Respective F proteins from different CDV strains are also available in publicly accessible databases of genetic information.
  • the F protein comprises an amino acid sequence that is at least 90% identical to SEQ ID NO: 3, and can be in different embodiments, at least 91% identical, or at least 92% identical, or at least 93% identical, or at least 94% identical, or at least 95% identical, or at least 96% identical, or at least 97% identical, or at least 98% identical, or at least 99% identical, or 100% identical to SEQ ID NO: 3.
  • the H protein is at least 90% identical but less than 100% identical to SEQ ID NO: 3
  • at least half of the differing amino acids or at least 60%, or at least 70%, or at least 80%, or at least 90%, or at least 95%, or 100%, are conservative substitutions.
  • One way to create the modified live canine adenovirus type 2 (CAV-2) vector carrying an antigen from the Canine Distemper Virus is to recombinantly insert a nucleic acid sequence encoding the antigen from the Canine Distemper Virus according to any of the embodiments described above into the genome of the modified live CAV-2 vector.
  • the nucleic acid sequence encoding the antigen from the Canine Distemper Virus comprises SEQ ID NO: 2 or, in view of the redundancies in the genetic code, an equivalent of SEQ ID NO: 2.
  • the nucleic acid sequence encoding the antigen from the Canine Distemper Virus comprises SEQ ID NO: 4 or, in view of the redundancies in the genetic code, an equivalent of SEQ ID NO: 4.
  • the methods of recombinantly inserting foreign nucleic acids into viral vectors are well known in the art.
  • the nucleic acid sequence encoding the antigen from the Canine Distemper Virus is inserted into the E3 region of the genome of the modified live CAV-2 vector by any of the methods well known in the art.
  • the nucleic acid sequence encoding the antigen from the Canine Distemper Virus be under operative control of a promoter.
  • Suitable promoters should be able to direct expression of the CDV antigen in the cells of the intended host, such as for example, canines.
  • Suitable promoters include, without limitations, SV-40 promoter, CMV promoter, RSV promoter, and other promoters known in the art.
  • the modified live CAV-2 vector carrying the CDV antigen according to any embodiments described herein is not enterically coated.
  • Enteric coating generally protects viruses from the harsh environment of the stomach, but applicants have previously discovered that enteric coating of a modified live CDV inhibited the immune response to oral vaccination with the modified live CDV.
  • the recombinant oral vaccine may comprise modified live canine parvovirus and/or modified live canine parainfluenza virus.
  • the suitable modified live CPV and CPI viruses are known in the art.
  • the respective CPV and CPI viruses present in VANGUARD® DAPPi vaccine may be administered orally even though VANGUARD® DAPPi vaccine is currently approved for parenteral administration.
  • VANGUARD® DAPPi vaccine In certain embodiments of the recombinant oral vaccine, neither of the modified live CPV and CPI is enterically coated.
  • the methods of manufacturing the viruses according to any embodiments described herein are known in the art. Without limitations, the viruses may be propagated on NLDK (Norden Lab Dog Kidney) cells as per standard procedures.
  • canine viruses can be grown in roller bottles with or without Gentamicin in Dulbecco’s Minimum Essential Media (DMEM) with 0-5% FBS.
  • DMEM Minimum Essential Media
  • NLDK cells are planted at an initial density of 1.2 x 10 4 to 3.5 x 10 4 cells / cm 2 and after 4-7 days of growth will be 80-100% confluent at the time of infection with virus.
  • the cells are infected with virus at a multiplicity of infection (MOI) between 0.001 and 1.0 tissue culture infective dose 50 (TCID 50 ), incubated at 34.0 – 38.0 °C, rotated at 0.2 -0.5 rpm, harvested after 2-5 days, and stored at less than or equal to -40°C.
  • MOI multiplicity of infection
  • TCID 50 tissue culture infective dose 50
  • the recombinant oral vaccines described herein elicit protective immune responses against the respective pathogens if administered orally.
  • the recombinant oral vaccines according to any embodiments described herein and containing the modified live CAV-2 vector carrying the CDV antigen elicit protective immune responses against CAV-1 infection, against CAV-2 infection, and against CDV infection.
  • the recombinant oral vaccine further comprises modified live CPV, then the recombinant oral vaccine also elicits a protective immune response against canine parvovirus.
  • the recombinant oral vaccine further comprises modified live CPI, then the recombinant oral vaccine also elicits a protective immune response against canine parainfluenza virus.
  • “Protective immune response” against CDV refers to reduction in duration or intensity of one or more clinical signs of CDV infection including, without limitations, CDV-caused mortality. Most preferably, the protective immune response against CDV refers to at least 95% (19 of 20) survival rate among the vaccinated dogs without showing any clinical signs of said disease in a challenge model wherein 80% (4 of 5) of the non-vaccinated dogs die of the disease, and 100% of the non-vaccinates show clinical signs after challenge.
  • “Protective immune response” against CAV-1 refers to reduction in duration or intensity of one or more clinical signs of CAV-1 infection including, without limitations, CDV-caused mortality.
  • the protective immune response against CAV-1 refers to at least 95% hepatitis in a challenge model wherein 80% of the non-vaccinated dogs do show clinical signs.
  • “Protective immune response” against CAV-2 refers to reduction in duration or intensity of one or more clinical signs of CAV-2 infection including, without limitations, CDV-caused mortality.
  • the protective immune response against CAV-2 refers to a significant difference in clinical signs between vaccinated and non-vaccinated dogs, in a challenge model wherein at least 60% of the non-vaccinated dogs show clinical signs of the disease.
  • “Protective immune response” against CPV refers to reduction in duration or intensity of one or more clinical signs of CPV infection including, without limitations, CDV-caused mortality.
  • the protective immune response against CPV refers to at least 95% (19 of 20) survival rate of the vaccinated dog during the observation period without showing more than one of the of following criteria of infection: temperature of ⁇ 103.4, lymphopenia of ⁇ 50 percent of pre-challenge normal, mucus or blood in the feces, and hemagglutination levels ⁇ 1:64 in a 1:% dilution of feces, in a challenge model wherein more than 80% of non-vaccinated dogs exhibit at least three of the above signs.
  • the protective immune response against CPI refers to a significant reduction in virus isolation (shedding) in the vaccinates as compared to non-vaccinated animals.
  • the recombinant oral vaccines may elicit the protective immune response after two oral administrations according to the first aspect of the invention or maintain (or elicit) the protective immune response after an annual revaccination according to the second aspect of the invention.
  • the parenteral administration is not needed to elicit the protective immune response against the target pathogen, as described above.
  • the second oral dose of the recombinant oral vaccine follows the first oral dose by 7 to about 35 days, or by 14 to 35 days or by 7 to 28 days, or by 14 to 28 days or by about 21 days.
  • the second oral dose of the recombinant oral vaccine may be followed by a third oral dose, wherein the third oral dose by 7 to about 35 days, or by 14 to 35 days or by 7 to 28 days, or by 14 to 28 days or by about 21 days.
  • the animals that receive the recombinant oral vaccine (whether in two or three oral doses) according to the first aspect of the invention have not been actively vaccinated against CDV prior to receiving the first oral dose of CDV via the recombinant oral vaccine described herein.
  • the methods further entail annual revaccinations of the recombinant oral vaccines accomplished via an oral administration of any of the vaccines disclosed herein, about one year after the first administration or a preceding annual oral administration of the vaccine, as would be understood by one of ordinary skill in the art.
  • the recombinant oral vaccines described herein may be used for revaccinations, e.g., annual revaccinations that occur about one year from administering first and second dose of a different vaccine and/or about one year from administering a previous annual revaccination, either with the vaccine described herein or the different vaccine.
  • the term “different vaccine” is interchangeable with the term “initial vaccine” and refers to vaccines containing the same antigen(s) as the vaccine according to the invention (in this case, at least a CDV antigen and a CAV-2 antigen) but not in the form of a recombinant CAV-2 vector comprising the CDV antigen, e.g., CDV H gene.
  • VANGUARD® line e.g., VANGUARD® DAP containing modified live CAV-2 virus, modified live CDV virus, and modified live CPV virus
  • vaccines of RECOMBITEC ® line e.g., RECOMBITEC ® C3 or RECOMBITEC ® C4
  • NOBIVAC® line e.g., NOBIVAC® CANINE 1- DAPPv
  • the puppy does not need to be MDA-negative and can be vaccinated with the initial vaccine as early as 4 weeks of age, as instructed by the manufacturer of the respective initial vaccines.
  • the dog undergoes the annual revaccination comprising orally administering to said dog the recombinant oral vaccine according to any embodiment described above.
  • modified live viruses in the recombinant oral vaccines may be present in the following amounts per dose (whether the first dose, the second dose, the optional third dose according to the first aspect, or the annual revaccination dose according to the first or the second aspect): modified live CAV-2 carrying the CDV antigen according to any embodiment of the invention: from 10 2.5 TCID 50 to about 10 8 TCID 50 , more preferably, from 10 3 TCID 50 to about 10 8 TCID 50 , more preferably, from 10 3 TCID 50 to about 10 6 TCID 50 , from 10 3 TCID 50 to about 10 5.5 TCID 50 , from 10 3 TCID 50 to about 10 5 TCID 50 , from 10 3 TCID 50 to about 10 4.5 TCID 50 , from about 10 3.5 TCID 50 to about 10 5.5 TCID 50 , from 10 3.5 TCID 50 to about 10 5 TCID 50 , from 10 3.5 TCID 50 to about 10 4.5 TCID 50 , from 10
  • the recombinant oral vaccines described herein may contain other components, including without limitations pharmaceutically acceptable excipients, including carriers, solvents, and diluents, isotonic agents, buffering agents, stabilizers, preservatives, immunomodulatory agents (e.g., interleukins, interferons, and other cytokines), vaso-constrictive agents, antibacterial agents, antifungal agents, and the like.
  • pharmaceutically acceptable excipients including carriers, solvents, and diluents, isotonic agents, buffering agents, stabilizers, preservatives, immunomodulatory agents (e.g., interleukins, interferons, and other cytokines), vaso-constrictive agents, antibacterial agents, antifungal agents, and the like.
  • Typical carriers, solvents, and diluents include water, saline, dextrose, ethanol, glycerol, and the like.
  • a pharmaceutically acceptable carrier includes any and all solvents, dispersion media, coatings, adjuvants, stabilizing agents, diluents, preservatives, antibacterial and antifungal agents, isotonic agents, adsorption delaying agents, and the like.
  • the carrier(s) must be "acceptable” in the sense of being compatible with the components of the invention and not deleterious to the subject to be immunized.
  • the carriers will be sterile and pyrogen- free, and selected based on the mode of administration to be used. It is well known by those skilled in the art that the preferred formulations for the pharmaceutically accepted carrier which comprise the vaccines are those pharmaceutical carriers approved in the applicable regulations promulgated by the United States (US) Department of Agriculture, or equivalent government agency in a non-US country. Therefore, the pharmaceutically accepted carrier for commercial- production of the vaccines is a carrier that is already approved or will be approved by the appropriate government agency in the US or foreign country.
  • the recombinant oral vaccine compositions optionally may include vaccine-compatible pharmaceutically acceptable (i.e., sterile and non-toxic) liquid, semisolid, or solid diluents that serve as pharmaceutical vehicles, excipients, or media.
  • Diluents can include water, saline, dextrose, ethanol, glycerol, and the like.
  • Isotonic agents can include sodium chloride, dextrose, mannitol, sorbitol, and lactose, among others.
  • Stabilizers include albumin, among others.
  • the recombinant oral vaccines according to any of the embodiments of the first or the second aspect may be non-adjuvanted.
  • the recombinant oral vaccines may further comprise an adjuvant.
  • Suitable adjuvants include, without limitations, salts of N-(2- Deoxy-2-L-leucylamino-b-D-glucopyranosyl)-N-octadecyldodecanoylamide, e.g., acetate salt, alone or in combination with other adjuvanting compounds such lecithin and DDA (dimethyl dioctadodecyl ammonium bromide), or PAM3CSK4.
  • the recombinant oral vaccines disclosed herein may further contain mucoadhesives.
  • Suitable mucoadhesives include, without limitations natural or synthetic hydrophilic substances which have organic functional groups (carboxyl, hydroxyl, and amino groups) or hydrogen bonds.
  • Some known mucoadhesive polymers are carbomers, cellulose derivatives, alginates, lectins, and tiolated polymers.
  • Specific mucoadhesives include CARBOPOPL® (a lightly cross-linked polyacrylic acid (PAA)), chitosan.
  • the recombinant oral vaccines may be administered in the doses having volume of between 0.25 and about 5 ml, e.g., between bout 0.5 ml and about 3 ml, or about 0.5 and 2 ml, or about 1 ml.
  • the vaccines are given to dogs that are MDA-negative and are about eight weeks of age or older (e.g., about 10 weeks, about 12 weeks, about 14 weeks, etc).
  • the first dose would be given to a dog at about four weeks of age (or about six weeks, about eight weeks, about 10 weeks, about 12 weeks, about 14 weeks, etc.), followed by subsequent doses according to the timetable given herein.
  • the recombinant oral vaccine is administered in the annual revaccination dose, then at the time of the annual revaccination the dog can be as young as about 14 months old (if the puppy had to wait until until it is MDA negative, and only then was orally vaccinated according to the methods of the first aspect of the invention).
  • CAV-2 vector carrying CDV H gene The recombinant Canine Adenovirus type 2 expressing the hemagglutinin (H) protein of canine distemper virus (rCAV-2::CDV-H) was generation by an initial transfection of the virulent Manhattan strain of CAV-2 with a plasmid containing a mCherry expression cassette and flanking sequences of gene E3 of the CAV-2 genome (rCAV-mC) cultured in Norden Lab dog kidney (NLDK) cells.
  • H hemagglutinin
  • NLDK Norden Lab dog kidney
  • the expression plasmid pCAV-SV40-H that was used to replace mCherry and generate CAV-SV40-H was created by cloning the SV40 promoter from pSV40 into NotI-AscI sites of pCAV- CMV-H by digestion and ligation.
  • transfer plasmid pCAV-CMV-mC was linearized by PacI restriction enzyme digestion.
  • the linearized plasmid was transfected into NLDK cells with Lipofectamine 3000 (Life Technologies) and infected with parental CAV-2 virus in 6-well plate.
  • the transfected/infected cells were harvested and subject to 4 plaque purifications for red fluorescent plaques by agarose overlay in 6-well plates.
  • One purified fluorescent plaque was further tested by PCR to confirm there was no wild-type CAV virus contamination. This plaque was scaled up in a T-75 flask.
  • CAV-mC The cells and supernatant of the T-75 flask culture was harvested, aliquoted and stored at -80o C freezer. This recombinant virus was designated as CAV-mC.
  • CAV-mC the SV40 promoter from pSV40 was cloned into the NotI-AscI sites of pCAV-CMV-H by digestion and ligation. Transfer plasmid pCAV-SV40-H was linearized by PacI restriction enzyme digestion. The linearized plasmid was transfected into NLDK cells with Lipofectamine 3000 and infected with CAV-mC virus in a 6-well plate.
  • the transfected/infected cells were harvested and subject to 4 plaque purifications for non-fluorescent plaques by agarose overlay in 6-well plates. Seven purified non- fluorescent plaques were further PCR confirmed to contain SV40-H insert. [0064] Three purified plaques were further passed/expanded, the whole cultures were harvested and stored at -80 C freezer. These 3 plaques were designated as CAV-SV40-H clone 1, clone 2, and clone 3. The passage 3 of the 3 clones were further subjected to sequencing confirmation of the inserts. PCR amplified insert region (SV40-H) of each clone was confirmed to have the correct sequences.
  • SV40-H PCR amplified insert region
  • CAV-SV40-H clone was selected as a candidate to advance forward to pre-master seed virus bank. This clone was further expanded through two passages (p4 and p5) by infection of NLDK cells cultured in DMEM medium containing 2 mM L-glutamine, and 10 ⁇ g/mL gentamicin to scale up for pre-MSV. Passage 5 material was then inoculated onto NLDK cells cultured in DMEM medium containing L glutamine and 0,05% gentamicin.
  • the pre-MSV was designated “CAV-SV40-H #1 p6 Lot 221202-011.” P6 indicates 6 passages for scaling up (after clonal purification).
  • the pre-MSV was frozen by storage at -80°C. [0066]
  • the CAV-SV40-H pre-MSV candidate Lot 221202-011 was tested for purity, potency, and identity. Purity was tested using the Modified Sterility/Purity test method and Mycoplasma using the Sigma Aldrich Lookout Mycoplasma qPCR Detection kit, ref: MP0400A-1KT. Potency and identity were tested by IFA method on CAV-SV40H infected NLDK cells.
  • potency and identity was determined by IFA method on CAV-SV40-H infected NLDK cells.
  • CAV-SV40-H pre-MSV Lot 221202-011 was titrated on NLDK to determine potency.
  • Wells containing CPE were tested for CAV-2 using goat polyclonal antibody specific for CAV-2 as the primary antibody and detected by a bovine anti-goat–IgG Alex Fluor 488 probe (green fluorescence).
  • the CDV-H identity was determined using canine polyclonal antibody as the primary antibody and detected by a rabbit anti-dog labeled with Cy3 probe (red fluorescence).
  • Example 2 Generation of CAV-2 virus expressing CDV F-protein.
  • the materials and methods in this example are similar to those disclosed in Example 1.
  • the nucleic acid sequence of CDV F protein (SEQ ID NO: 4).
  • pCAV-CMV2-F that contains CDV F gene under a truncated CMV promoter was synthesized by GenScript.
  • Transfer plasmid pCAV- CMV2-F was linearized by PacI restriction enzyme digestion.
  • the linearized plasmid was transfected into NLDK cells with Lipofectamine 3000 (Life Technologies) and infected with CAV- mC virus in 6-well plate.
  • the transfected/infected cells were harvested and subject to 4 plaque purifications for non-fluorescent plaques by agarose overlay in 6-well plates.
  • Six non-fluorescent plaques from the 4 th plaque purification were picked.
  • PCR of N and C-terminus of CMV2-F insert showed all were positive.
  • Three of the plaques were passed in T-25 flask for 3 times and stored at -80 o C freezer.
  • the 3 recombinant viruses were designated as CAV-CMV2-F clone 1, clone 2, clone 3.
  • Example 3 DNAs of the passage 3 viral cultures were extracted, and PCR was performed to confirm the stability.
  • Example 3 Canine Adenovirus Type 2 carrying CDV antigens confers protective response to CDV challenge.
  • the objective of the study was to assess the efficacy of two modified live vaccines when administered orally in dogs against a Canine Distemper Virus challenge administered intravenously (IV) at a dilution of 1:50 approximately three weeks post-third vaccination.
  • the vaccines utilized a recombinant canine adenovirus-2 (rCAV-2) expressing the canine distemper virus (CDV) H or F protein.
  • Anti-parasitic and/or antibiotic treatments were administered at the supplier site according to their procedures. Animals received additional pretreatments as determined by ARS Veterinarians as needed upon arrival for up to three (3) days.
  • Serology samples (approximately 6 mL) were collected in SST tubes from all animals on Days 0, 21, and 42 (prior to each vaccination) Day 63 (prior to challenge), and 84.
  • Nasal swabs were collected using polyester swabs and sterile collection tubes containing approximately 3.0 mL of Viral Transport Media (VTM supplemented with antibiotics) prior to Day 0.
  • Fecal swabs were collected (one swab per 3.0 mL VTM tube) prior to Day 0.
  • Clinical Observations were recorded on Day -1, twice on Day 0 (prior to vaccination and 3-6 hours post-vaccination), once daily on Days 1-7 and 20, twice on Day 21 (prior to vaccination and 3-6 hours post-vaccination), once daily on Days 22-28 and 41, twice daily on Day 42 (prior to vaccination and 3-6 hours post- vaccination), and once daily on Days 43-49 during the vaccination phase.
  • Clinical Observations were recorded on Day 62, twice on Days 63-68, three times on Days 69-75, twice on Days 76-83 and one on Day 84 during the challenge phase.
  • Tympanic temperatures were recorded once on Day -1, twice on Day 0 (prior to vaccination and 3-6 hours post-vaccination), once daily on Days 1-7 and 20, twice on Day 21 (prior to vaccination and 3-6 hours post-vaccination), once daily on Days 22-28 and 41, twice daily on Day 42 (prior to vaccination and 3-6 hours post-vaccination), and once daily on Days 43-49 during the vaccination phase.
  • Tympanic temperatures were recorded once on Day 62, twice on Day 63 (prior to challenge and approximately 3-6 hours post-challenge), and at least once daily Days 64 to 84 during the challenge phase. Post-challenge, the veterinarians considered the humane endpoints of the study.
  • the T02 group receiving the CAV-2::CDV-H construct orally, never showed clinical signs of distemper post-challenge.
  • Table 4 Frequency Distributions of Clinical Signs of Disease Post-Challenge by Treatment Group Present Post Challenge? No Yes Total No. of No. of No. of Clinical Sign Treatment Animals % Animals % Animals T01 9 90.00 1 10.00 10 Agitation T02 10 100.00 0 0.00 10 T03 10 100.00 0 0.00 10 T01 2 20.00 8 80.00 10 Anorexia T02 10 100.00 0 0.00 10 T03 10 100.00 0 0.00 10 T01 10 100.00 0 0.00 10 Cough T02 10 100.00 0 0.00 10 T03 10 100.00 0 0.00 10 Present Post Challenge? No Yes Total No.
  • Geometric Means Summary of CDV Titers by Treatment Group Day of Number of Geometric Treatment Study Animals Mean Minimum Maximum -1 10 1.0 1.0 1.0 1.0 20 10 1.0 1.0 1.0 41 10 1.0 1.0 1.0 1.0 T01 62 10 1.0 1.0 1.0 1.0 71 1 1.0 1.0 1.0 72 7 1.0 1.0 1.0 84 2 304.4 256.0 362.0 T02 -1 10 1.0 1.0 1.0 1.0 Day of Number of Geometric Treatment Study Animals Mean Minimum Maximum 20 10 4.2 1.0 23.0 41 10 29.9 11.0 64.0 62 10 59.7 23.0 181.0 84 10 1782.8 512.0 2896.0 -1 10 1.0 1.0 1.0 1.0 20 10 1.9 1.0 11.0 T03 41 10 38.1 16.0 91.0 62 10 84.3 11.0 256.0 84 10 1722.1 1024.0 2896.0 Table 6.
  • Geometric Means Summary of CAV-1 Titers by Treatment Group Day of Number of Geometric Treatment Study Animals Mean Minimum Maximum -1 10 1.0 1.0 1.0 1.0 20 10 1.0 1.0 1.0 41 10 1.0 1.0 1.0 1.0 T01 62 10 5.5 5.5 5.5 71 1 5.5 5.5 5.5 72 6 5.5 5.5 5.5 84 2 5.5 5.5 5.5 -1 10 1.0 1.0 1.0 1.0 20 10 208.2 64.0 512.0 T02 41 10 831.7 362.0 2048.0 62 10 776.0 256.0 2048.0 84 10 891.4 362.0 2048.0 -1 10 1.0 1.0 1.0 1.0 1.0 20 10 111.7 23.0 256.0 T03 41 10 461.4 181.0 1024.0 62 10 675.5 362.0 1448.0 84 10 512.0 181.0 1024.0 Table 7.
  • Geometric Means Summary of CAV-2 Titers by Treatment Group Day of Number of Geometric Treatment Study Animals Mean Minimum Maximum -1 10 1.0 1.0 1.0 1.0 20 10 1.0 1.0 1.0 41 10 1.0 1.0 1.0 1.0 T01 62 10 1.0 1.0 1.0 1.0 71 1 1.0 1.0 1.0 72 7 1.0 1.0 1.0 84 2 1.0 1.0 1.0 1.0 -1 10 1.0 1.0 1.0 1.0 20 10 54.1 1.0 512.0 T02 41 10 446.7 23.0 1448.0 62 10 187.2 11.0 1448.0 84 10 1217.7 512.0 2896.0 -1 10 1.0 1.0 1.0 1.0 1.0 20 10 87.8 3.0 362.0 T03 41 10 461.4 64.0 1024.0 62 10 362.0 32.0 1448.0 84 10 724.0 362.0 1448.0 [0087] All dogs (100%) were negative to CDV, CAV-1, and CAV-2 on Day -1.

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Abstract

La présente divulgation concerne un procédé de protection d'un canidé contre le virus de la maladie de Carré, le procédé comprenant l'administration d'un vaccin oral recombinant comprenant un vecteur de type 2 d'adénovirus canin (CAV-2) vivant modifié portant un antigène du virus de la maladie de Carré (virus CDV) audit canidé, le vaccin étant administré par voie orale.
PCT/US2024/052376 2023-10-24 2024-10-22 Vaccins oraux canins et méthodes d'administration Pending WO2025090476A1 (fr)

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Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998000166A1 (fr) * 1996-07-03 1998-01-08 Merial, Inc. ADENOVIRUS CANIN DE RECOMBINAISON (ACR) CONTENANT DE l'ADN EXOGENE
US20100028379A1 (en) 2006-12-27 2010-02-04 Pfizer, Inc Methods of vaccine administration
US20100291549A1 (en) 2006-04-27 2010-11-18 Pikamab, Inc. Methods and compositions for antibody therapy
WO2018057441A1 (fr) * 2016-09-20 2018-03-29 Boehringer Ingelheim Vetmedica Gmbh Vecteurs d'adénovirus canin

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998000166A1 (fr) * 1996-07-03 1998-01-08 Merial, Inc. ADENOVIRUS CANIN DE RECOMBINAISON (ACR) CONTENANT DE l'ADN EXOGENE
US20100291549A1 (en) 2006-04-27 2010-11-18 Pikamab, Inc. Methods and compositions for antibody therapy
US20100028379A1 (en) 2006-12-27 2010-02-04 Pfizer, Inc Methods of vaccine administration
WO2018057441A1 (fr) * 2016-09-20 2018-03-29 Boehringer Ingelheim Vetmedica Gmbh Vecteurs d'adénovirus canin

Non-Patent Citations (8)

* Cited by examiner, † Cited by third party
Title
APPEL MJ ET AL., VET REC, vol. 105, 1979, pages 156 - 159
APPEL, MJ, ADV VET MED., vol. 41, 1999, pages 309 - 324
BASS EP ET AL., JAVMA, vol. 177, 1980, pages 234 - 242
CLIQUET FLORENCE ET AL: "Oral vaccination of dogs: a well-studied and undervalued tool for achieving human and dog rabies elimination", VETERINARY RESEARCH, vol. 49, no. 1, 13 July 2018 (2018-07-13), London, UK, XP093240488, ISSN: 1297-9716, Retrieved from the Internet <URL:http://link.springer.com/content/pdf/10.1186/s13567-018-0554-6.pdf> DOI: 10.1186/s13567-018-0554-6 *
FISCHER L ET AL: "Vaccination of puppies born to immune dams with a canine adenovirus-based vaccine protects against a canine distemper virus challenge", VACCINE, ELSEVIER, AMSTERDAM, NL, vol. 20, no. 29-30, 4 October 2002 (2002-10-04), pages 3485 - 3497, XP004381813, ISSN: 0264-410X, DOI: 10.1016/S0264-410X(02)00344-4 *
MOUZIN DE, IAVMA, vol. 224, 2004, pages 55 - 60
SWANGO LJ., NORDEN NEWS, vol. 58, 1983, pages 4 - 10
ZHANG ET AL: "Oral vaccination of dogs (Canis familiaris) with baits containing the recombinant rabies-canine adenovirus type-2 vaccine confers long-lasting immunity against rabies", VACCINE, ELSEVIER, AMSTERDAM, NL, vol. 26, no. 3, 3 December 2007 (2007-12-03), pages 345 - 350, XP022419013, ISSN: 0264-410X, DOI: 10.1016/J.VACCINE.2007.11.029 *

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