Mutated non-primate lentiviral Env proteins and their use as drugs

Patent metadata

Field	Value
Publication number	US-9974852-B2
Application number	US-201414896505-A
Country	US
Kind code	B2
Filing date	Jun 6, 2014
Priority date	Jun 7, 2013
Publication date	May 22, 2018
Grant date	May 22, 2018

Abstract

A pharmaceutical composition includes, as active substance a mutated non-primate lentiviral Env protein having decreased immunosuppressive properties, substantially no immunosuppressive properties or no immunosuppressive properties, or a variant of the mutated lentiviral Env protein, or a fragment of the above proteins, in association with a pharmaceutically acceptable carrier.

First claim

The invention claimed is:

1. A pharmaceutical composition comprising as an active substance an isolated mutated feline lentiviral ENV protein that has lost at least 50%, at least 75% or 100% of its immunosuppressive activity in comparison to immunosuppressive activity of a wild type ENV protein, or a fragment thereof, said fragment of said isolated mutated feline lentiviral ENV protein comprising at least 40 amino acids, said mutated feline lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type feline lentiviral ENV protein, said mutated ENV protein having at least 80% identity to the sequence SEQ ID NO: 5, said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:

(SEQ ID NO: 1)

A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A,

wherein,
X 1 is A or R, and X 2 , X 3 , X 4 and X 5 are any amino acid, or
X 2 is A or R, and X 1 , X 3 , X 4 and X 5 are any amino acid, or
X 3 is A or R, and X 1 , X 2 , X 4 and X 5 are any amino acid, or
X 4 is A or R, and X 1 , X 2 , X 3 and X 5 are any amino acid, or
X 5 is A or R, and X 1 , X 2 , X 3 and X 4 are any amino acid,

in association with a pharmaceutically acceptable carrier.

2. A pharmaceutical composition comprising as an active substance an isolated mutated feline lentiviral ENV protein that has lost at least 50%, at least 75% or 100% of its immunosuppressive activity in comparison to immunosuppressive activity of a wild type ENV protein, or a fragment thereof, said fragment of said isolated mutated feline lentiviral ENV protein comprising at least 40 amino acids, said mutated feline lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of a wild type feline lentiviral ENV protein, said mutated ENV protein having at least 80% identity to the sequence SEQ ID NO: 5, said mutated feline lentiviral ENV protein or fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:

(SEQ ID NO: 3)

[V/I]-[E/R]-A-[I/M/T/L]-X1-X2-X3-X4-X5-T-A-[F/L]-

A-M,

wherein,
X 1 is A or R, and X 2 , X 3 , X 4 and X 5 are any amino acid, or
X 2 is A or R, and X 1 , X 3 , X 4 and X 5 are any amino acid, or
X 3 is A or R, and X 1 , X 2 , X 4 and X 5 are any amino acid, or
X 4 is A or R, and X 1 , X 2 , X 3 and X 5 are any amino acid, or
X 5 is A or R, and X 1 , X 2 , X 3 and X 4 are any amino acid,

in association with a pharmaceutically acceptable carrier.

3. The pharmaceutical composition according to claim 1 , wherein
said loss of at least 50%, at least 75% or 100% of the immunosuppressive activity of said mutated feline lentiviral ENV protein or of said fragment thereof being liable to be assessed by the fact that in an in vivo assay involving engrafted tumor cells rejection, in animals excluding human beings,
said tumor cells being transduced either so as to express said mutated ENV protein or said fragment (mutated ENV tumor cells),
or said tumor cells being transduced so as to express said wild type ENV protein or a fragment thereof (wild type ENV tumor cells),
or said tumor cells being not transduced (normal tumor cells),
the following ratio:
immunosuppression index of said mutated ENV protein or of said fragment (i mutated env )/immunosuppression index of wild type ENV protein (i wild type env ) is less than 0.5,
i mutated env being defined by: (maximum area reached by mutated ENV tumor cells−maximum area reached by normal tumor cells)/(maximum area reached by normal tumor cells), and
i wild type env being defined by: (maximum area reached by wild type ENV tumor cells−maximum area reached by normal tumor cells)/(maximum area reached by normal tumor cells).

4. The pharmaceutical composition according to claim 1 , wherein:
X 1 is A or R, X 2 is A or R, and X 3 , X 4 and X 5 are any amino acid, or
X 1 is A or R, X 3 is A or R, and X 2 , X 4 and X 5 are any amino acid, or
X 1 is A or R, X 4 is A or R, and X 2 , X 3 and X 5 are any amino acid, or
X 1 is A or R, X 5 is A or R, and X 2 , X 3 and X 4 are any amino acid, or
X 2 is A or R, X 3 is A or R, and X 1 , X 4 and X 5 are any amino acid, or
X 2 is A or R, X 4 is A or R, and X 1 , X 3 and X 5 are any amino acid, or
X 2 is A or R, X 5 is A or R, and X 1 , X 3 and X 4 are any amino acid, or
X 3 is A or R, X 4 is A or R, and X 1 , X 2 and X 5 are any amino acid, or
X 3 is A or R, X 5 is A or R, and X 1 , X 2 and X 4 are any amino acid, or
X 4 is A or R, X 5 is A or R, and X 1 , X 2 and X 3 are any amino acid.

5. The pharmaceutical composition according to claim 1 , wherein:
X 1 is R, and X 2 , X 3 , X 4 and X 5 are any amino acid, or
X 2 is R, and X 1 , X 3 , X 4 and X 5 are any amino acid, or
X 3 is R, and X 1 , X 2 , X 4 and X 5 are any amino acid, or
X 4 is R, and X 1 , X 2 , X 3 and X 5 are any amino acid, or
X 5 is R, and X 1 , X 2 , X 3 and X 4 are any amino acid.

6. The pharmaceutical composition according to claim 1 , wherein:
X 1 is R, X 2 is R, and X 3 , X 4 and X 5 are any amino acid, or
X 1 is R, X 3 is R, and X 2 , X 4 and X 5 are any amino acid, or
X 1 is R, X 4 is R, and X 2 , X 3 and X 5 are any amino acid, or
X 1 is R, X 5 is R, and X 2 , X 3 and X 4 are any amino acid, or
X 2 is R, X 3 is R, and X 1 , X 4 and X 5 are any amino acid, or
X 2 is R, X 4 is R, and X 1 , X 3 and X 5 are any amino acid, or
X 2 is R, X 5 is R, and X 1 , X 3 and X 4 are any amino acid, or
X 3 is R, X 4 is R, and X 1 , X 2 and X 5 are any amino acid, or
X 3 is R, X 5 is R, and X 1 , X 2 and X 4 are any amino acid, or
X 4 is R, X 5 is R, and X 1 , X 2 and X 3 are any amino acid.

7. The pharmaceutical composition according to claim 1 , wherein:
X 1 is A or R, and X 2 , X 3 , X 4 and X 5 are any amino acid different from A, G or R, or
X 2 is A or R, and X 1 , X 3 , X 4 and X 5 are any amino acid different from A, G or R, or
X 3 is A or R, and X 1 , X 2 , X 4 and X 5 are any amino acid different from A, G or R, or
X 4 is A or R, and X 1 , X 2 , X 3 and X 5 are any amino acid different from A or R, or
X 5 is A or R, and X 1 , X 2 , X 3 and X 4 are any amino acid different from A, G or R.

8. The pharmaceutical composition according to claim 1 , wherein:
X 1 is A or R, X 2 is A, G or R, and X 3 , X 4 and X 5 are any amino acid different from A, G or R, or
X 1 is A or R, X 3 is A, G or R, and X 2 , X 4 and X 5 are any amino acid different from A, G or R, or
X 1 is A or R, X 4 is A, G or R, and X 2 , X 3 and X 5 are any amino acid different from A, G or R, or
X 1 is A or R, X 5 is A, G or R, and X 2 , X 3 and X 4 are any amino acid different from A, G or R, or
X 2 is A or R, X 3 is A, G or R, and X 1 , X 4 and X 5 are any amino acid different from A, G or R, or
X 2 is A or R, X 4 is A, G or R, and X 1 , X 3 and X 5 are any amino

Assignees

• Viroxis S A S
• Centre Nat Rech Scient
• Roussy Inst Gustave
• Univ Paris Sud Xi
• Viroxis Sas

Inventors

• Heidmann Thierry

Classifications

• A61K2039/53

  DNA (RNA) vaccination · CPC title

• C07K14/005

  from viruses · CPC title

• A61K39/21 Primary

  Retroviridae, e.g. equine infectious anemia virus · CPC title

• A61K2039/572

  cytotoxic response · CPC title

• C12N2740/15034

  Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein · CPC title