WO2021003421A1 - Suppression du virus de tumeur mammaire - Google Patents

Suppression du virus de tumeur mammaire Download PDF

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Publication number
WO2021003421A1
WO2021003421A1 PCT/US2020/040742 US2020040742W WO2021003421A1 WO 2021003421 A1 WO2021003421 A1 WO 2021003421A1 US 2020040742 W US2020040742 W US 2020040742W WO 2021003421 A1 WO2021003421 A1 WO 2021003421A1
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WO
WIPO (PCT)
Prior art keywords
mtv
dosage
protein
vaccine
administering
Prior art date
Application number
PCT/US2020/040742
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English (en)
Inventor
Sophie Bartsich
Original Assignee
Virago Vax Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from US16/460,295 external-priority patent/US12029783B2/en
Application filed by Virago Vax Inc. filed Critical Virago Vax Inc.
Priority to EP20834735.1A priority Critical patent/EP3993873A4/fr
Priority to CA3145293A priority patent/CA3145293A1/fr
Publication of WO2021003421A1 publication Critical patent/WO2021003421A1/fr

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Classifications

    • 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
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • 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
    • A61K2039/80—Vaccine for a specifically defined cancer
    • A61K2039/812—Breast
    • 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
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/12011—Betaretrovirus, e.g. mouse mammary tumour virus
    • C12N2740/12022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
    • 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
    • C12N2740/00—Reverse transcribing RNA viruses
    • C12N2740/00011—Details
    • C12N2740/10011—Retroviridae
    • C12N2740/12011—Betaretrovirus, e.g. mouse mammary tumour virus
    • C12N2740/12034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein

Definitions

  • MTV MTV belongs to a group globally recognized as oncogenic. As such, it acts through insertional mutagenesis, ultimately producing a tumor in its target tissue.
  • a hormonally-activated and regulated retrovirus the genetic code of MTV is fully deciphered for variants found in some mammalian species.
  • the genetic code of the deciphered variants share a common genetic architecture with many other retroviruses with major expression groups comprising: long terminal repeats (LTR); group antigen polyproteins (GAG); reverse transcriptases and polymerases (POL); and, envelope proteins (ENV).
  • LTR long terminal repeats
  • GAG group antigen polyproteins
  • POL reverse transcriptases and polymerases
  • ENV envelope proteins
  • Figure 1 provides an illustration of this generalized architectural structure for MTV.
  • the proteins that compose the GAG group generally form the viral core structure, RNA genome binding proteins, and are the major proteins comprising the nucleoprotein core.
  • the GAG group may also encode viral matrix, capsid, and nucleoproteins.
  • the POL reverse transcriptase is the essential enzyme that carries out the reverse transcription process that takes the viral RNA genome to a double-stranded DNA pre integrated form.
  • the ENV group encodes for host cell transmembrane and surface-receptor subunits, and regulatory subunits found on the endoplasmic reticulum of the host cell in addition to the proteins that form the viral envelope.
  • HMTV shares homology with MMTV in both form and action and the two share similar genetic sequences.
  • “homolog” and its derivatives are, unless specified with a percentage, used as commonly understood by practitioners in the field of biochemistry in that proteins sharing similar behavioral properties, structural motifs, or performing similar functions with conserved domains across a family are said to be“homologous” to each other.
  • RNA segments known to originate from MMTV (Axel, 1962).
  • the HMTV virus inserts similarly into the genome at random sites, ultimately inciting comparable mutations by activating various oncogenes (Callahan, 2012).
  • Representatives of the envelope protein can be localized to the host cell membrane, but are more prominent in cellular cytoplasm (Tomana, 1981).
  • the retrovirus replication cycle is characterized by conversion of the single-strand
  • An embodiment of the invention is a method of reducing breast cancer recurrence via prevention of MTV transcription in breast tissue via administration of a protease inhibitor.
  • Other compositions contemplated as part of various embodiments of the instant invention provide a means for stabilizing or reducing the amount of virus present in a human or other animal.
  • a patient with a breast cancer diagnosis, or a patient with breast cancer risk factors is treated with an antiretroviral medication. In still other embodiments this treatment is combined with the administration of MTV transition proteins or transition protein peptides.
  • a cDNA encoding an MTV transition protein is provided. The protein may be myristoylated and, likewise, the cDNA may encode for this modification. Also provided are cDNA molecules encoding MTV transition protein peptide chains of greater or lesser length with or without a myristol. The transition protein or transition protein peptide chains may then be incorporated as part of an MTV vaccine or treatment regimen administered to a patient for control of MTV particle count or viral activity.
  • antigens may be combined in a single injection that can prevent different diseases or that protect against multiple strains of infectious agents causing the same disease (e.g. combination vaccine DPT combining diphtheria, pertussis, and tetanus antigens).
  • combination vaccines can be useful to overcome the logistic constraints of multiple injections and accommodate for a patient’s fear of needles and pain.
  • the term“polyvalent vaccine” refers to multiple antigen types that may derive from a single or multiple disease vectors whereas a“combination vaccine” refers to a vaccine with multiple antigens targeting at least two or more diseases.
  • a“polyvalent vaccine” may have multiple antigens but only provide immunity to a single disease whereas a “combination vaccine” may contain a single type of antigen from two or more disease vectors and give rise to an immunity for those two or more diseases.
  • the term “patient” reflects a human or animal subject to whom a medical treatment is provided either for modification of a disease state or as a prophylactic preventative of disease.
  • LAV live attenuated vaccines
  • pathogens virus or bacteria
  • Inactivated vaccines are made from microorganisms (viruses, bacteria, other) that have been killed through physical or chemical processes. These killed organisms cannot cause disease.
  • Subunit vaccines like inactivated whole-cell vaccines do not contain live components of the pathogen. They differ from inactivated whole-cell vaccines, by containing only the antigenic parts of the pathogen. Protein based subunit vaccines present an antigen to the immune system without viral particles, using a specific, isolated protein of the pathogen. Some bacteria when infecting humans are often protected by a polysaccharide (sugar) capsule that helps the organism evade the human defense systems especially in infants and young children. Polysaccharide vaccines create a response against the molecules in the pathogen’s capsule. Conjugate subunit vaccines also create a response against the molecules in the pathogen’ s capsule. In comparison to plain polysaccharide vaccines, they benefit from a technology that binds the polysaccharide to a carrier protein that can induce a long-term protective response.
  • Toxoid vaccines are based on the toxin produced by certain bacteria (e.g. tetanus or diphtheria).
  • the toxin invades the bloodstream and is largely responsible for the symptoms of the disease.
  • a protein-based toxin is rendered harmless (toxoid) and used as the antigen in the vaccine to elicit immunity.
  • the toxoid is often adsorbed to aluminum or calcium salts, which serve as adjuvants.
  • an“syn” such as synMTV or synENV, synPOL, etc. to differentiate from naturally occurring versions.
  • an synENV protein may be derived from a genetic sequence identical to or modified from (e.g., introns removed, different starter code and reading frame, different promoters etc.) a naturally occurring sequence but produced in an alternative system of protein expression (e.g., in yeast or e. coli , etc.).
  • the amino acid sequences, and underlying genetics, for each protein may vary across viral sub-species found in different mammalian species.
  • synProteins or synProtein subunits may be derived in whole or in part from single or multiple viral sub-types (e.g., human sequence synProtein combined with a homologous canine synProtein to form a polyvalent vaccine protective against both HMTV and canine MTV virus particles).
  • an engineered vaccine is created by synthesizing and purifying an EP3-pl4-plO peptide chain.
  • the peptides, in haptan formulation, are coupled to keyhole limpet hemocyanin (KLH) and extensively dialyzed. Moles of conjugated peptide/mole of KLH are determined by amino acid analysis and range between 700 and 1,000. Positive and negative controls consist of gp52, purified from C3H-MMTV and a scrambled peptide sequence coupled to KLH respectively.
  • the MTV envelope (ENV) class of proteins is known to have conserved domain elements across variants of the virus detected across multiple species.
  • the ENV proteins may comprise those that are signaling peptides, those that are specific to the outer membrane of the virus particle, those specific to the endoplasmic reticulum, and those that are trans-membrane anchors.
  • the polymerase (POL) protein class may comprise both a polymerase and/or a reverse transcriptase and an associated signaling factor.
  • the group antigen (GAG) proteins form the viral core structure, RNA genome binding proteins, and are the major proteins comprising the nucleoprotein core particle. The POL and GAG group proteins are likewise thought to have conserved cross-species elements.
  • MTV proteins are synthesized with at least two major precursor polyproteins, for MMTV this includes: gPr75env containing gp52 and gp36; and, Pr75gag containing p27, pp20, pl4, and plO.
  • SEQ. ID: 01 illustrates an example sequence for a Pr75gag precursor polyprotein, the cleavage products of which are schematically illustrated in Figure 2. Shown in Figure 2, for example, pl4 is a signal peptide subunit of an ENV protein. pl4 can also function in both oncogenic and an anti-oncogenic capacity depending on its phosphorylation status.
  • RNA viruses tend to be unstable but highly virulent
  • DNA viruses tend to be stable but not as aggressive.
  • DNA and RNA viruses can work together, each building upon the strengths of the other to increase pathogenicity. It is believed that many viruses like HPV and HSV can make other viruses more virulent, and that certain combinations are particularly effective at inducing oncogenesis in patients.
  • a monovalent vaccine targeting a rabbit may use an MTV protein or protein particles purified from rabbit-specific MTV.
  • a human targeting vaccine may use ENV protein purified from HMTV strains found in humans.
  • a synthetically derived synProteins or hybrids may be created based upon a target species MTV genetic sequence.
  • MTV derived POL or GAG proteins or protein subunits may be used.
  • Protease inhibitors are an example of one class of retroviral medications. As described above, MTV reproduction relies upon production and then cleavage of precursor proteins. In general, a protease inhibitor prevents cleavage of the viral precursor protein by inhibiting the action of the viral protease responsible for the cleavage. Thus, viral loads in a host are decreased as the virus fails to replicate. Accordingly, an embodiment of the invention results in the decrease of MTV loads in a patient accomplished through the administration of protease inhibitors.
  • ritonavir may be provided to a patient as a maintenance dosage in the range of 50-440 mg per day, or atazanvir at 300-400 mg per day.
  • ritonavir may be used with a range of 50-400 mg per day combined with another protease inhibitor at 100-400 mg per day.
  • Protease inhibitors may also be used in combination with chemotherapy drugs to first treat a cancer (e.g., breast cancer or other viral-mediated cancer) and then used singly to prevent cancer recurrence.
  • dosage ranges explicitly contemplate every integer or part dosage amount (e.g., 1, 1.1, 1.2, 1.3, etc.) within the range including the initial and end amounts within the range. It is recognized that while ranges may list start and end points manufacturing tolerances and the like may result in products above or below range within a specified tolerance.
  • a method of treating MTV exposure in a patient can comprise: determining the integer or part dosage amount (e.g., 1, 1.1, 1.2, 1.3, etc.) within
  • the method described above may further be modified wherein the administration of the protease inhibitor is performed at a first treatment dosage and a second maintenance dosage.
  • an MTV vaccine may be administered during the treatment dosage administration.
  • the MTV vaccine may be administered when switching to a maintenance dosage.
  • the maintenance dosage is in the range of 50-200 mg of medication per day.
  • the ritonavir is combined with at least one of: atazanavir (Reyataz), darunavir (Prezista), fosamprenavir (Lexiva), indinavir (Crixivan), lopinavir/ritonavir (Kaletra), nelfmavir (Viracept), ritonavir (Norvir), saquinavir (Invirase), tipranavir (Aptivus), atazanavir/cobicistat (Evotaz), and, darunavir/cobicistat (Prezcobix).
  • Normal dosage amounts may vary from approximately 1 to 100,000 micrograms, up to a total dose of about 10 grams, depending upon the route of administration. Desirable dosages include 250 pg, 500 pg, 1 mg, 50 mg, 100 mg, 150 mg, 200 mg, 250 mg, 300 mg, 350 mg, 400 mg, 450 mg, 500 mg, 550 mg, 600 mg, 650 mg, 700 mg, 750 mg, 800 mg, 850 mg, 900 mg, 1 g, 1.1 g, 1.2 g, 1.3 g, 1.4 g, 1.5 g, 1.6 g, 1.7 g, 1.8 g, 1.9 g, 2 g, 3 g, 4 g, 5 g, 6 g, 7 g, 8 g, 9 g, and 10 g ⁇
  • Imidazole competes with the his-tag for binding to the metal-charged resin and thus is used for elution of the protein from an IMAC column.
  • a low concentration of imidazole is added to both binding and wash buffers to interfere with the weak binding of other proteins and to elute any proteins that weakly bind.
  • His-tagged protein is then eluted with a higher concentration of imidazole. Proteins with different numbers of poly histidine tags elute differently from nickel -affinity resin.
  • HN-tag alternating histidine and asparagine
  • HAT-tag which is a peptide tag
  • myristoylated transition protein peptide chain is 90% or more homologous with the peptide encoded by SEQ ID No: 2. In still other embodiments the myristoylated transition protein peptide chain is 10 or more contiguous amino acids as found in SEQ ID No: 2 or an acceptably homologous sequence.
  • sequence explicitly contemplates DNA, cDNA, RNA and resulting peptide chains encoded thereby in both sense and antisense directions. To know one is to know the others via the standard rules of complementarity and codon encoding as exemplified in standardized DNA, RNA, and amino acid codon tables.

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Virology (AREA)
  • Veterinary Medicine (AREA)
  • Animal Behavior & Ethology (AREA)
  • Chemical & Material Sciences (AREA)
  • Public Health (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Oncology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Communicable Diseases (AREA)
  • Molecular Biology (AREA)
  • Immunology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Epidemiology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Abstract

L'invention concerne des méthodes de traitement d'un patient exposé à un virus de tumeur mammaire (VTM). L'invention concerne également un ADNc purifié codant pour une chaîne peptidique de protéine de transition VTM. L'invention concerne également un vaccin contenant une protéine de transition VTM. Une autre version de l'invention concerne un vaccin comprenant des polypeptides VTM couplés à une protéine porteuse. Le VTM peut être traité par la fourniture d'un vaccin VTM avec une protéine de transition VTM ; et l'administration dudit vaccin. Dans d'autres cas, le vaccin VTM est administré avec un médicament antirétroviral.
PCT/US2020/040742 2019-07-02 2020-07-02 Suppression du virus de tumeur mammaire WO2021003421A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
EP20834735.1A EP3993873A4 (fr) 2019-07-02 2020-07-02 Suppression du virus de tumeur mammaire
CA3145293A CA3145293A1 (fr) 2019-07-02 2020-07-02 Suppression du virus de tumeur mammaire

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US16/460,295 US12029783B2 (en) 2019-07-02 Mammary tumor virus vaccine
US16/460,295 2019-07-02
US201962934858P 2019-11-13 2019-11-13
US62/934,858 2019-11-13

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WO2021003421A1 true WO2021003421A1 (fr) 2021-01-07

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12016844B2 (en) 2019-07-02 2024-06-25 Virago Vax Inc. Mammary tumor virus suppression

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WO1999051268A1 (fr) * 1998-04-08 1999-10-14 International Bioimmune Systems, Inc. Compositions immunomodulatoires d'antigenes mmtv et leurs techniques d'application
US20070281911A1 (en) * 2004-01-21 2007-12-06 Gilead Sciences Inc. Use of Adefovir or Tenofovir for Inhibiting Mmtv-Like Viruses Involved in Breast Cancer and Primary Biliary Cirrhosis
US20120135950A1 (en) * 2009-05-21 2012-05-31 The Regents Of The University Of Michigan Antiviral treatment of lymphoma and cancer
US9383367B1 (en) * 2010-12-07 2016-07-05 Chunli Liu Methods of detecting conjugation site-specific and hidden epitope/antigen
WO2017147078A1 (fr) * 2016-02-22 2017-08-31 Brigham Young University Méthode et compositions pour le traitement de l'anémie par inhibition de furine
US20190343938A1 (en) * 2019-07-02 2019-11-14 Virago Vax Inc. Mammary tumor virus vaccine

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WO1999051268A1 (fr) * 1998-04-08 1999-10-14 International Bioimmune Systems, Inc. Compositions immunomodulatoires d'antigenes mmtv et leurs techniques d'application
US20070281911A1 (en) * 2004-01-21 2007-12-06 Gilead Sciences Inc. Use of Adefovir or Tenofovir for Inhibiting Mmtv-Like Viruses Involved in Breast Cancer and Primary Biliary Cirrhosis
US20120135950A1 (en) * 2009-05-21 2012-05-31 The Regents Of The University Of Michigan Antiviral treatment of lymphoma and cancer
US9383367B1 (en) * 2010-12-07 2016-07-05 Chunli Liu Methods of detecting conjugation site-specific and hidden epitope/antigen
WO2017147078A1 (fr) * 2016-02-22 2017-08-31 Brigham Young University Méthode et compositions pour le traitement de l'anémie par inhibition de furine
US20190343938A1 (en) * 2019-07-02 2019-11-14 Virago Vax Inc. Mammary tumor virus vaccine

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12016844B2 (en) 2019-07-02 2024-06-25 Virago Vax Inc. Mammary tumor virus suppression

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Publication number Publication date
CA3145293A1 (fr) 2021-01-07
EP3993873A1 (fr) 2022-05-11
EP3993873A4 (fr) 2023-09-20

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