US 2003/0158131 A1Published application

DNA vectors containing mutated HIV proviruses

AssigneeChildren's Medical Center Corporation

InventorAnna Aldovini

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Abstract

The present invention pertains to mutated, non-infectious HIV viral particles, vectors for production of such particles and vaccines employing such vectors. The non-infectious particles are obtained by introducing a number of inactivating mutations into a native viral genome. These mutations are designed so as to minimize the probability of genetic reversion to an infectious virus, while retaining the basic protein content and immunogenic properties of a wild-type virion. The altered viral genome expresses proteins that can assemble into non-infectious particles which contain immunogenic components of the virus, but which are unable to infect cells. The preferred mutations are introduced in at least one amino acid position of the NC protein in combination with at least one other mutation in an amino acid position of the RT protein or the In protein. In one embodiment, the mutations to the native HIV genome may also be made in at least one amino acid position of the NC protein, at least one position in the RT protein, and at least one position in the In protein. In another embodiment, the mutations to the native HIV genome may be introduced in clusters, where two or more mutations are made in the NC protein, the RT protein, the In protein, or any combinations thereof.

Claims41

Dependent claims

A nucleic acid construct which encodes non-infectious HIV particles, the construct comprising:

a) at least one mutation in the gag gene of the HIV genome that encodes the amino acid sequence of the nucleocapsid (NC) protein, wherein the mutation is made in at least one position selected from the group consisting of the 5′ flanking region (amino acids 1-14 of SEQ ID NO:
8), the 5′ Cys-His box (amino acids 15-28 of SEQ ID NO:
8), the peptide linker region (amino acids 29-35 of SEQ ID NO:
8), the 3′ Cys-His box (amino acids 36-49 of SEQ ID NO:
8) and the 3′ flanking region (amino acids 50-55 of SEQ ID NO:
8), such that the mutation prevents viral RNA packaging;
and
b) at least one further mutation in the pol gene selected from the group consisting of
(i) at least one mutation in the region of the pol gene that encodes the amino acid sequence in the finger domain (amino acid 1 to 84 of SEQ ID NO:
4) of the reverse transcriptase (RT) protein, such that a dysfunctional reverse transcriptase protein is produced to prevent viral RNA conversion to DNA and
(ii) at least one mutation in the region of the pol gene that encodes the amino acid sequence of the catalytic (amino acids 51-212 of SEQ ID NO:
6) or multimerization domain (amino acids 1-50 of SEQ ID NO:
6) of the Integrase (In) protein, such that dysfunctional Integrase protein is produced to prevent a transcribed viral DNA molecule from integrating with a host cell.

10 more elements

Dependencies are inferred from the claim text.

Facts

Application: 10/000,511Filed 23 Oct 2001
Priority: 23 Oct 2000
Published: 21 Aug 2003
Term ends: Not estimatedThe source records don’t support an estimate.

Classification16 codes

C12N 15/86
For animal cells: viral vectors
A61K 39/12
Medicinal preparations containing antigens or antibodies: viral antigens
A61K 39/21
Viral antigens: retroviridae, e.g. equine infectious anemia virus
C07K 14/005
Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof: from viruses