EP1000157A1 - Compositions renfermant des capsomeres de papillomavirus humain homogene, leurs procedes de production et leurs procedes d'utilisation en tant qu'agents diagnostiques, prophylactiques ou therapeutiques - Google Patents

Compositions renfermant des capsomeres de papillomavirus humain homogene, leurs procedes de production et leurs procedes d'utilisation en tant qu'agents diagnostiques, prophylactiques ou therapeutiques

Info

Publication number
EP1000157A1
EP1000157A1 EP98933101A EP98933101A EP1000157A1 EP 1000157 A1 EP1000157 A1 EP 1000157A1 EP 98933101 A EP98933101 A EP 98933101A EP 98933101 A EP98933101 A EP 98933101A EP 1000157 A1 EP1000157 A1 EP 1000157A1
Authority
EP
European Patent Office
Prior art keywords
hpv
capsomeres
stable
protein
vlps
Prior art date
Legal status (The legal status 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 status listed.)
Withdrawn
Application number
EP98933101A
Other languages
German (de)
English (en)
Inventor
Joann A. Suzich
Michael P. Mccarthy
Robert C. Rose
Robert L. Garcea
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
MedImmune LLC
University of Technology Corp
University of Rochester
Original Assignee
MedImmune LLC
University of Technology Corp
University of Rochester
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
Application filed by MedImmune LLC, University of Technology Corp, University of Rochester filed Critical MedImmune LLC
Publication of EP1000157A1 publication Critical patent/EP1000157A1/fr
Withdrawn legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
    • 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
    • A61P31/20—Antivirals for DNA 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
    • 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
    • C12N2710/20011—Papillomaviridae
    • C12N2710/20022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • the present invention relates to stable human
  • HPV papillomavirus
  • the present invention further relates to modi ⁇
  • Infection is typi- cally characterized by the induction of benign epithelial
  • Each species of vertebrate is infected by a
  • canine and rabbit papillomaviruses cannot induce
  • HPV types 6 and 8 are prevalent sexually transmitted disease.
  • lesions are benign, lesions arising from certain papillo ⁇
  • mavirus types e.g., HPV-16 and HPV-18, can undergo malig ⁇
  • HPV-16 is the most common, being found in about
  • L2 the minor capsid protein
  • HPVs and COPV Similar to HPVs associ ⁇
  • LI major capsid protein
  • virus-like particles using a
  • papillomavirus major capsid proteins See, e.g., Hines et
  • VLPs virus-like particles
  • VLPs have been reported to be morphologically
  • HPV major capsid protein containing compositions that
  • HPV capsomeres which present conformational epitopes
  • cation comprises a carboxy deletion and/or at least one
  • modified HPV LI nucleic acid sequences which comprise a carboxy-terminal deletion and/or substitution
  • the invention generally relates to stable
  • HPV capsomeres which present at least one virus-neutraliz- ing conformational epitope expressed by an LI protein
  • HPVs In particular, they react with conformational
  • compositions As the subject capsomeres, unlike non- modified LI proteins, do not give rise to virus-like
  • compositions according to the invention should be sub ⁇
  • PV avirus
  • VLPs are morphologically and antigenically simi ⁇
  • VLPs may be produced in vivo
  • suitable host cells e.g., mammalian and insect host
  • capsomeres comprise the "monomer" units which constitute
  • Disulfide bond formation may be
  • VLPs are then "capped off", e.g., by reaction with alkyl ⁇
  • ating agents e.g., iodoacetamide or N-ethylmaleimide .
  • the subject stable capsomeres will express at least one
  • mavirus which is comprised of capsomeres. More specifi ⁇
  • cally it is constituted of seventy-two capsomeres in a
  • T 7 icosahedron structure.
  • the subject stable papillomavirus capsomeres which is also expressed by an LI protein of a corresponding native
  • epitopes is essential to the efficacy (both as prophylac-
  • HPV LI protein immunogens tic and diagnostic agents
  • LI protein e.g., major capsid protein expressed on the
  • VLPs when expressed by suitable host cells e.g., mammali-
  • the modification will prevent VLP
  • VLPs mined by known methods, e.g., by visual detection of VLPs
  • HPV LI DNA may further comprise
  • assembly will comprise carboxy-terminal deletions and/or removal of at least one cysteine residue that inhibits or
  • this is effected by deletion and/or
  • cysteine residue e.g., to sterically hinder the cysteine
  • cysteine residues involved in VLP assembly can be
  • cysteine residue (s) are comprised
  • the protein was mixed with sample preparation
  • HPV-11 preparations were treated at 4°C as
  • VLPs incubated with 10 mM DTT, 5mM EDTA for 16 hours.
  • VLPs in PBS were incubated with 5% ⁇ ME (a), or 200 mM NaHC0 3 , pH 9.6(b) for 16 hours at 4°C
  • VLPs treated as described
  • VLPs disassembled in the presence of 200 mM
  • ucts were separated on 2% agarose gels. Gels were stained
  • actin band is 0.6 kb.
  • Lane A contains molecular size
  • Lanes B and C contain PCR products obtained with RNA isolated from cells infected with HPV-11 pre-incubated
  • lane F and G are from cells incubated without
  • Lanes H-L contain
  • capsid antiserum (10 ⁇ 3 -10 ⁇ 7 ) . This antiserum neutralizes
  • Lanes M-Q show PCR
  • the present invention generally relates
  • the present invention was based, in part, on obser-
  • residues in the C-terminal end of the LI protein may be any residues in the C-terminal end of the LI protein.
  • capsomeric bonds which stabilize the capsid, by extending
  • the present invention was further based on experi-
  • portion of the LI protein inhibits (or prevents) VLP
  • the first method comprises ex ⁇
  • the second method comprises expression of a non-
  • the second method will comprise the following
  • VLP reassembly e.g., by
  • LI DNA or a fragment thereof as a hybridization probe LI DNA or a fragment thereof as a hybridization probe
  • the HPV LI DNA said in the subject inven ⁇ is cloned and expressed.
  • the HPV LI DNA said in the subject inven ⁇ is cloned and expressed.
  • cancer or condyloma acuminata e.g., HPV-16, HPV-18, HPV-
  • HPV-33 HPV-35, HPV-39, HPV-45, HPV-51, HPV-52, and
  • HPV-56 are involved in cancer, and HPV-6, HPV-11, HPV-30,
  • HPV-42, HPV-43, HPV-44, HPV-54, HPV-55,and HPV-70, are examples of HPV-42, HPV-43, HPV-44, HPV-54, HPV-55,and HPV-70, are examples of HPV-42, HPV-43, HPV-44, HPV-54, HPV-55,and HPV-70, are examples of HPV-42, HPV-43, HPV-44, HPV-54, HPV-55,and HPV-70, are examples
  • HPV LI DNA may be produced from any desired HPV LI DNA.
  • the selected HPV LI DNA will be modified such that the
  • oligomers such as VLPs and which present at least one
  • boxy-terminal deletions can be introduced by cleaving the
  • LI deletions can be constructed by PCR mutagenesis
  • the LI gene can be made by DNA synthesis.
  • the selected HPV LI DNA will be mutagenized by
  • substitution modification i.e., substituting a selected
  • cysteine residues targeted for modification are those at
  • cysteine residue targeted for modification is found at
  • disulfide bond formation may be pre-
  • HPV-L1 sequence may impart synergistic effects. Moreover, as noted in this embodiment of the invention, the deletion
  • the selected host and expression vector will be any suitable host and expression vector.
  • an intracellular expression vector is
  • the host cells will need to be lysed and the HPV
  • expression vector contains sequences that facilitate
  • HPV capsomeres can be recovered directly from
  • system comprises the baculovirus/insect cell system used
  • HPV LI proteins can be produced in other cells.
  • promoters e.g., promoters, polyadenylation sequences, enhancers,
  • selectable markers are also well known. The selection of
  • HPV LI DNAs the host cells should only express HPV LI
  • This aspect of the invention will preferably be
  • VLPs VLPs, and the conversion of said VLPs into stable capso ⁇
  • the object of the present invention is to
  • the expressed HPV LI proteins can be any suitable amino acid sequence.
  • the expressed HPV LI proteins can be any suitable amino acid sequence.
  • reducing agent e.g., on the order of 1% to 5% by
  • ing agents such as iodoacetamide after reduction.
  • PV types known in the art Further, particular types of PVs are associated with particular infections such as flat
  • HPV type determines, in part, the site of infection
  • Virus particles can also be isolated for a particular
  • papillomavirus can be isolated, the amino acid sequence
  • HPV capsomere Since the HPV capsomere must express at least one
  • the expression system will comprise a vector
  • baculovirus vectors are preferably
  • a baculovirus system offers the advantage that
  • baculovirus is an insect virus
  • glycosylation and phosphorylation which may be
  • Baculovirus vector systems are known in the art.
  • infected cells express HPV LI proteins exhibiting appro ⁇
  • an LI gene or modified LI gene is operably linked into an
  • the signals may be derived from signals from sources.
  • tion include the cloning of the LI gene into an expression
  • Example 6 an in vitro assay suitable for determining
  • capsomeres according to the invention may be any type of serotyping, capsomeres according to the invention.
  • phosphate dehydrogenase triose phosphate isomerase
  • peroxidase alkaline phosphatase
  • asparaginase glucose
  • radioisotopic labels examples include 3 H,
  • nescent labels include a luminal label, an isoluminal
  • luciferin label a luciferin label, an luciferase label, an aequorin label
  • Well-known carriers include glass, polystyrene,
  • polypropylene polyethylene, dextran, nylon, amylases,
  • the nature of the carrier can be either
  • the vaccines of the invention will contain an
  • vaccines may be effected by any pharmaceutically accept ⁇
  • able means e.g., parenterally, locally or systemically,
  • the vaccine may be any suitable human papillomavirus.
  • the vaccine may be any suitable human papillomavirus.
  • immunologically acceptable carrier is preferably used
  • saline such as saline or phosphate-buffered saline.
  • the vaccines will be administered in therapeutically.
  • the vaccines will be administered in dosages ranging from
  • multiple dosages can be administered.
  • the method of the present invention makes possible
  • PV type As more than one PV type may be associated with PV
  • the vaccines may comprise stable HPV capso ⁇
  • HPV 16 and 18 are associated with cervical carcinomas
  • a vaccine for cervical neoplasia may comprise
  • HPVs 3a and 3a are associated with PV infections.
  • HPVs 3a and 3a are associated with PV infections.
  • HPVs 3a and 3a are associated with PV infections.
  • EV modysplasia verruciformis
  • HPVs 1, 2, 4, and 7 have been reported to be associated with cutaneous warts and HPVs 6b, 11a, 13, and
  • subject vaccine formulations may comprise a mixture of
  • HPV capsomeres of the invention can be any HPV capsomeres of the invention.
  • kits will comprise the
  • the anti-HPV mouse monoclonal antibody AU1 was pur ⁇
  • Hll.Fl and H11.A3 were purchased from Pennsylvania State
  • Mg 2+ was from Gibco/BRL, ECL reagents were purchased
  • HPV-11 LI proteins were heterologously expressed in
  • Trichoplusia ni High Five ® cells infected with recom ⁇
  • binant baculovirus encoding the complete LI open reading
  • the clarified lysate was
  • VLPs were diluted > 2 -fold in PBS-0.5M
  • sucrose (w/w in PBS -0.5M NaCl) .
  • the gradients were
  • VLPs were then dialyzed into selected buffers (either PBS,
  • pellet (typically none was visible) was resuspended in 100 ⁇ l of IX Laemmli sample preparation buffer.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Virology (AREA)
  • General Health & Medical Sciences (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Public Health (AREA)
  • General Chemical & Material Sciences (AREA)
  • Oncology (AREA)
  • Communicable Diseases (AREA)
  • Biotechnology (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Veterinary Medicine (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Peptides Or Proteins (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

La présente invention concerne des capsomères du HPV stables qui expriment au moins un épitope de conformation neutralisant le virus d'une protéine L1 native du HPV et qui sont sensiblement incapables de s'assembler en particules de type viral. Ces capsomères, en raison de leur taille plus petite et de leurs propriétés immunogènes, conviennent bien à une utilisation dans des vaccins anti-HPV et comme agents diagnostiques. De plus, en raison de leur taille plus petite (par rapport aux particules de type viral), ces capsomères stables peuvent être facilement purifiés et devraient permettre d'obtenir des vaccins anti-HPV à homogénéité accrue.
EP98933101A 1997-07-03 1998-07-02 Compositions renfermant des capsomeres de papillomavirus humain homogene, leurs procedes de production et leurs procedes d'utilisation en tant qu'agents diagnostiques, prophylactiques ou therapeutiques Withdrawn EP1000157A1 (fr)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
US88805097A 1997-07-03 1997-07-03
US888050 1997-07-03
PCT/US1998/013799 WO1999001557A1 (fr) 1997-07-03 1998-07-02 Compositions renfermant des capsomeres de papillomavirus humain homogene, leurs procedes de production et leurs procedes d'utilisation en tant qu'agents diagnostiques, prophylactiques ou therapeutiques

Publications (1)

Publication Number Publication Date
EP1000157A1 true EP1000157A1 (fr) 2000-05-17

Family

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Family Applications (1)

Application Number Title Priority Date Filing Date
EP98933101A Withdrawn EP1000157A1 (fr) 1997-07-03 1998-07-02 Compositions renfermant des capsomeres de papillomavirus humain homogene, leurs procedes de production et leurs procedes d'utilisation en tant qu'agents diagnostiques, prophylactiques ou therapeutiques

Country Status (5)

Country Link
EP (1) EP1000157A1 (fr)
JP (1) JP2002510976A (fr)
AU (1) AU755679B2 (fr)
CA (1) CA2295316C (fr)
WO (1) WO1999001557A1 (fr)

Families Citing this family (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5437951A (en) 1992-09-03 1995-08-01 The United States Of America As Represented By The Department Of Health And Human Services Self-assembling recombinant papillomavirus capsid proteins
US6228368B1 (en) * 1997-10-06 2001-05-08 Loyola University Of Chicago Papilloma virus capsomere formulations and method of use
US20020039584A1 (en) 1998-02-20 2002-04-04 Medigene Ag Papilloma virus capsomere vaccine formulations and methods of use
US6926897B1 (en) 1998-03-24 2005-08-09 Medigene Aktiengesellschaft Medicament for the avoidance or treatment of papillomavirus-specific tumour
AU3730600A (en) 1999-03-18 2000-10-04 Xiaojiang Chen Compositions preparations and uses of human papillomavirus l1 protein
US6245568B1 (en) 1999-03-26 2001-06-12 Merck & Co., Inc. Human papilloma virus vaccine with disassembled and reassembled virus-like particles
WO2003068933A2 (fr) * 2002-02-14 2003-08-21 Novavax, Inc. Optimisation de sequences geniques de particules pseudo-virales pour l'expression dans des cellules d'insectes
JP4549858B2 (ja) 2002-10-17 2010-09-22 バイオリーダーズ コーポレイション ヒト・パピローマウイルスに対するワクチン用ベクターおよび同ベクターによって形質転換された微生物
WO2006028214A1 (fr) 2004-09-10 2006-03-16 Asahi Glass Company, Limited Vaccin pour administration orale

Family Cites Families (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE122007000085I1 (de) * 1991-07-19 2008-03-27 Univ Queensland St Lucia Polynukleotidabschnitt des HPV16-Genoms
DE122007000098I1 (de) * 1992-06-25 2008-03-27 Papillomavirus vakzine
DK0809700T3 (da) * 1994-10-07 2006-09-18 Univ Loyola Chicago Papillomaviruslignende partikler, fusionsproteiner samt fremgangsmåde til fremstilling heraf

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO9901557A1 *

Also Published As

Publication number Publication date
CA2295316C (fr) 2007-06-26
WO1999001557A1 (fr) 1999-01-14
AU8284298A (en) 1999-01-25
CA2295316A1 (fr) 1999-01-14
JP2002510976A (ja) 2002-04-09
AU755679B2 (en) 2002-12-19

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