WO2005068634A1 - Vaccins de synthese du sous-type c indien contre le vih-1 a utiliser chez l'homme - Google Patents

Vaccins de synthese du sous-type c indien contre le vih-1 a utiliser chez l'homme Download PDF

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Publication number
WO2005068634A1
WO2005068634A1 PCT/IN2004/000284 IN2004000284W WO2005068634A1 WO 2005068634 A1 WO2005068634 A1 WO 2005068634A1 IN 2004000284 W IN2004000284 W IN 2004000284W WO 2005068634 A1 WO2005068634 A1 WO 2005068634A1
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WO
WIPO (PCT)
Prior art keywords
hiv
gene
genes
subtype
vaccine constructs
Prior art date
Application number
PCT/IN2004/000284
Other languages
English (en)
Inventor
Pradeep Seth
Original Assignee
Pradeep Seth
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 Pradeep Seth filed Critical Pradeep Seth
Publication of WO2005068634A1 publication Critical patent/WO2005068634A1/fr

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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
    • 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
    • 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
    • C12N2710/24011—Poxviridae
    • C12N2710/24111—Orthopoxvirus, e.g. vaccinia virus, variola
    • C12N2710/24141—Use of virus, viral particle or viral elements as a vector
    • C12N2710/24143—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vector
    • 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
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16111—Human Immunodeficiency Virus, HIV concerning HIV env
    • C12N2740/16122—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
    • C12N2740/16011—Human Immunodeficiency Virus, HIV
    • C12N2740/16311—Human Immunodeficiency Virus, HIV concerning HIV regulatory proteins
    • C12N2740/16322—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

Definitions

  • This invention relates to development of HIV-1 vaccine candidates and to the process of preparation there of.
  • this invention relates to Indian HIV-1 subtype C constructs comprising HIV-1 subtype C envelope (gp120) and capsld (gag-protease), nef and tat genes cloned in plasmid ONA vector, and viral vector.
  • AIDS is one of the most dreaded infections afflicting the human race today. Therefore, development of a safe and effective vaccine clearly is paramount to halting the AIDS epidemic. All living forms, particularly human beings, have an immune system, which protects the body against the invasion of various microorganisms including viruses. However, HIV invades the cells of immune system and destroys it. Consequently virus replicates in the body unchecked. An effective vaccine against HIV can halt this march of virus in human beings, which has been a goal for the scientific community. An effective vaccine should induce a long-lasting and robust immune response, which would pre-empt invasion of HIV in human body.
  • HIV vaccines such as use of attenuated live virus constructs or killed viruses pose significant safety issues with HIV, although these have succeeded for other virus infections like poliovirus or measles virus.
  • Alternate approach that has been employed for making HIV vaccines is Recombinant DNA technology. This approach has allowed scientists to use portions of HIV genes to induce immune response. These genes are mounted on a carrier DNA molecule called a vector and then injected into a recipient in whom 5 it induces an immune response against HIV; thus providing a high degree of safety. Similarly, these genes may also be mounted on a virus vector, which induce HIV-1 specific immune response in an animal when injected.
  • Ae object of thi ⁇ invention is to propose a vaccine against Indian subtype C of i o Human immunodeficiency Virus type 1 (HIV-1) for human beings.
  • Another object of this invention is to propose a vaccine for effectual combating HIV-1 subtype C virus infection.
  • HIV vaccine constructs based on recombinant DNA technology to combat HIV-1 subtype C infections, comprising human codon optimized structural genes (envelope and gag-protease) and non structural genes (nef and tat) of Indian strains of HIV-1 subtype C mounted on plasmid DNA vector and viral vector.
  • the structural genes, gp120 and gag-protease, of Indian HIV-1 subtype C are a part of nucleic acid of HIV-1 subtype C viruses.
  • the non-structural genes are required for the replication of the virus.
  • the sequence of these genes is modified to human codon frequency without changing the amino acid sequence.
  • the codon optimized structural genes are synthesized and then mounted on a plasmid DNA vector or a virus vector, which carry these genes to the cell inside the human body. These genes are then processed by the cell to produce proteins without affecting the cellular functions.
  • PCR Polymerase chain reaction
  • a reaction mix constituting of buffer, forward and reverse primers, dNTPs (dATP, dGTP, dCTP, dTTP), MgCI 2 and Taq DNA
  • the five amplified genes were sequenced by direct sequencing reaction using primer walking strategy. DNA of these amplified genes were purified by ethanol precipitation and sequencing reaction was put up for each DNA 5 preparation using the Big Dye terminator Kit and cycle sequencing was carried out in PCR System 2400. The sequence data obtained were then sorted, aligned and analysed. Thereafter, it was subjected to codon optimization as per human codon frequency manually.
  • the codon optimized genes were constructed by using the PCR amplification method. The constructed genes were then sequenced in an automated sequencer. The sequence data of the synthesized genes were then 0 compared with the expected sequence and both sequences had a 100% homology. The codon optimized genes were named as: env-gp 120- 29692CO, env-gp120-49426CO, gag-protease49587CO, IND-tatCO, IND-nefCO. 5. Cloning of Codon Optimized genes in plasmid vector
  • NK-29692CO, NK-49426CO and NK-49587CO, NK-IND- tatCO and NK-JND-nefCO, were then tested for in vitro expression in 293T cells
  • Prime boost strategy was employed to immunize Macaca radiata (bonnet monkeys). HIV-1 plasmid DNA vaccine constructs were used for priming the animals with single dose of recombinant plasmids constructs, NK-29692CO, NK-

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biophysics (AREA)
  • Biochemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Virology (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
  • Peptides Or Proteins (AREA)

Abstract

Cette invention se rapporte à des vaccins de synthèse contre le virus de l'immunodéficience humaine (VIH) pour combattre l'infection au VIH-1 du sous-type C indien chez l'homme, qui comprennent des gènes structurels et non structurels de la souche indienne du sous-type C du VIH-1 montés sur un vecteur de plasmide d'ADN.
PCT/IN2004/000284 2004-01-16 2004-09-10 Vaccins de synthese du sous-type c indien contre le vih-1 a utiliser chez l'homme WO2005068634A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
IN63/DEL/04 2004-01-16
IN63DE2004 2004-01-16

Publications (1)

Publication Number Publication Date
WO2005068634A1 true WO2005068634A1 (fr) 2005-07-28

Family

ID=34779367

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IN2004/000284 WO2005068634A1 (fr) 2004-01-16 2004-09-10 Vaccins de synthese du sous-type c indien contre le vih-1 a utiliser chez l'homme

Country Status (3)

Country Link
CN (1) CN1906301A (fr)
WO (1) WO2005068634A1 (fr)
ZA (1) ZA200605498B (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2116605A3 (fr) * 2004-06-17 2010-01-06 Wyeth Plasmide doté de trois unités transcriptionnelles complètes et compositions immunogènes pour induire une réponse immunitaire au VIH

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
DATABASE EMBASE [online] CHUGH P. ET AL: "Cloning of gag gene of HIV subtype (Indian strain) into a mammalian expression vector and in vitro expression studies.", Database accession no. (EMB-2003346217) *
DATABASE MEDLINE [online] GUPTA S. ET AL: "Gag-derived proteins of HIV isolates from Indian patients: cloning, expression, and purification of p24 of B and C subtypes.", Database accession no. (NLM10910720) *
INDIAN JOURNAL OF MEDICAL MICROBIOLOGY., vol. 21, no. 2, 2003, pages 77 - 81 *
PROTEIN EXPRESSION AND PURIFICATION., vol. 19, no. 3, pages 321 - 328 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2116605A3 (fr) * 2004-06-17 2010-01-06 Wyeth Plasmide doté de trois unités transcriptionnelles complètes et compositions immunogènes pour induire une réponse immunitaire au VIH
US8623382B2 (en) 2004-06-17 2014-01-07 Wyeth Llc Immunogenic compositions for inducing an immune response to HIV

Also Published As

Publication number Publication date
CN1906301A (zh) 2007-01-31
ZA200605498B (en) 2007-09-26

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