IP Library › Granted Patent US 9,636,396

Granted Patent B2

US 9,636,396 · App. 13/707,885 · Granted May 2, 2017

Mutant human and simian immunodeficiency virus ENV proteins with reduced immunosuppressive properties

Inventor: Thierry Heidmann (Paris, FR)

Assignees: CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE; UNIVERSITE PARIS-SUD XI; INSITUT GUSTAVE ROUSSY; VIROXIS S.A.S.

A61K39/21 A61K39/12 C07K14/005 C07K7/06 C07K7/08 C07K14/162 C12N2740/15022 C12N2740/15033 C12N2740/15034 C12N2740/16022 C12N2740/16034 C12N2740/16111 C12N2740/16122 C12N2740/16134

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Quick Facts

Patent No.: US 9,636,396
App. No.: 13/707,885
Granted: May 2, 2017
Kind: B2

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Abstract

A pharmaceutical composition includes, as active substance a mutated lentiviral ENV protein, substantially devoid of 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.

Claims (233)

1. A pharmaceutical composition comprising as active substance:

a) an isolated mutated human or simian lentiviral ENV protein having no or a reduced immunosuppressive activity as compared to a corresponding wild type non-mutated human or simian lentiviral ENV protein,

said mutated human or simian lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of the corresponding wild type non-mutated human or simian lentiviral ENV protein,

said mutated human or simian lentiviral ENV protein comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q,

(SEQ ID NO: 416)

wherein,

X represents any amino acid,

and either

X a is A, F, G, L or R, and X b is any amino acid,

X a is any amino acid, and X b is R, or

X a is A, F, G, L, R, and X b is R,

b) at least one fragment of said isolated mutated human or simian lentiviral ENV protein having no or a reduced immunosuppressive activity as compared to the corresponding wild type non-mutated human or simian lentiviral ENV protein,

said fragment comprising at least 40 amino acids,

said fragment comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q,

(SEQ ID NO: 416)

wherein,

X represents any amino acid,

and either

X a is A, F, G, L or R, and X b is any amino acid,

X a is any amino acid, and X b is R, or

X a is A, F, G, L, R, and X b is R,

in association with a pharmaceutically acceptable carrier, said absence or reduction of immunosuppressive activity of the above mentioned mutated human or simian lentiviral ENV protein or of the above defined fragment being liable to be assessed by the fact that in an in vivo assay involving engrafted tumor cells rejection,

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 the corresponding wild type non-mutated 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).

2. The pharmaceutical composition according to claim 1 comprising as active substance:

a) an isolated mutated human or simian lentiviral ENV protein having no or a reduced immunosuppressive activity as compared to the corresponding wild type non-mutated human or simian lentiviral ENV protein,

said mutated human or simian lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of the corresponding wild type non-mutated human or simian lentiviral ENV protein,

said mutated human or simian lentiviral ENV protein comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q,

(SEQ ID NO: 416)

wherein,

X represents any amino acid,

and either

X a is A, F, G, L or R, and X b is L, I, V, M or P,

X a is Y, I, H, C or T, and X b is R, or

X a is A, F, G, L or R, and X b is R,

said fragment comprising at least 40 amino acids,

said fragment comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q,

(SEQ ID NO: 416)

wherein,

X represents any amino acid,

and either

X a is A, F, G, L or R, and X b is L, I, V, M or P,

X a is Y, I, H, C or T, and X b is R, or

X a is A, F, G, L or R, and X b is R,

in association with a pharmaceutically acceptable carrier.

3. The pharmaceutical composition according to claim 1 comprising as active substance:

said mutated human or simian lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of the corresponding wild type non-mutated human or simian lentiviral ENV protein,

said mutated human or simian lentiviral ENV protein comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q,

(SEQ ID NO: 416)

wherein

X represents any amino acid,

and either

X a is A, F, G, L or R, and X b is L,

X a is Y, and X b is R, or

X a is A, F, G, L, or R, and X b is R,

said fragment comprising at least 40 amino acids,

said fragment comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q,

(SEQ ID NO: 416)

wherein

X represents any amino acid,

and either

X a is A, F, G, L or R, and X b is L,

X a is Y, and X b is R, or

X a is A, F, G, L, or R, and X b is R,

in association with a pharmaceutically acceptable carrier.

4. The pharmaceutical composition according to claim 1 comprising as active substance:

said mutated human or simian lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of the corresponding wild type non-mutated human or simian lentiviral ENV protein,

said mutated human or simian lentiviral ENV protein comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q,

(SEQ ID NO: 416)

wherein

X represents any amino acid,

and either

X a is A, F, G, L or R, and X b is L, or

X a is Y, and X b is R,

said fragment comprising at least 40 amino acids,

said fragment comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q,

(SEQ ID NO: 416)

wherein

X represents any amino acid,

and either

X a is A, F, G, L or R, and X b is L, or

X a is Y, and X b is R,

in association with a pharmaceutically acceptable carrier.

5. The pharmaceutical composition according to claim 1 comprising as active substance:

said mutated human or simian lentiviral ENV protein resulting from mutation of the transmembrane (TM) subunit of the corresponding wild type non-mutated human or simian lentiviral ENV protein,

said mutated human or simian lentiviral ENV protein comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q (SEQ ID NO: 416),

wherein,

X represents any amino acid,

and either

X a is A, G or R, and X b is L, or

X a is Y, and X b is R,

said fragment comprising at least 40 amino acids,

said fragment comprising a mutated immunosuppressive domain (ISU) containing the following amino acid sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q,

(SEQ ID NO: 416)

wherein,

X represents any amino acid,

and either

X a is A, G or R, and X b is L, or

X a is Y, and X b is R,

in association with a pharmaceutically acceptable carrier.

6. The pharmaceutical composition according to claim 1 ,

wherein

X a is R, and X b is L, or

X a is Y, and X b is R.

7. The pharmaceutical composition according to claim 1 , wherein said isolated mutated human or simian lentiviral ENV protein or said fragment of said isolated mutated human or simian lentiviral ENV protein comprises one of the following amino acid sequences

A-I-E-K-X a -X b -X-DQ

(SEQ ID NO: 422),

A-I-E-R-X a -X b -X-DQ

(SEQ ID NO: 423),

A-V-E-K-X a -X b -X-DQ

(SEQ ID NO: 424),

A-V-E-R-X a -X b -X-DQ

(SEQ ID NO: 425).

8. The pharmaceutical composition according to claim 1 , wherein said isolated mutated human or simian lentiviral ENV protein or said fragment of said isolated mutated human or simian lentiviral ENV protein comprises one of the amino acid sequences:

SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 17, SEQ ID NO: 25, SEQ ID NO: 29, SEQ ID NO: 33, SEQ ID NO: 37, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 46, SEQ ID NO: 50, SEQ ID NO: 54, SEQ ID NO: 58, SEQ ID NO: 62, SEQ ID NO: 66, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 75, SEQ ID NO: 79, SEQ ID NO: 83, SEQ ID NO: 87, SEQ ID NO: 91, and SEQ ID NO:95.

9. The pharmaceutical composition according to claim 1 , wherein said isolated mutated human or simian lentiviral ENV protein or said fragment of said isolated mutated human or simian lentiviral ENV protein comprises one of the amino acid sequences:

SEQ ID NO: 9, SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 17, SEQ ID NO: 38, SEQ ID NO: 39, SEQ ID NO: 40, SEQ ID NO: 41, SEQ ID NO: 42, SEQ ID NO: 46, SEQ ID NO: 67, SEQ ID NO: 68, SEQ ID NO: 69, SEQ ID NO: 70, SEQ ID NO: 71 and SEQ ID NO: 75.

10. The pharmaceutical composition according to claim 1 , wherein said isolated mutated human or simian lentiviral ENV protein or said fragment of said isolated mutated human or simian lentiviral ENV protein comprises one of the amino acid sequences:

SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 104, SEQ ID NO: 108, SEQ ID NO: 112, SEQ ID NO: 116, SEQ ID NO: 120, SEQ ID NO: 124, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 133, SEQ ID NO: 137, SEQ ID NO: 141, SEQ ID NO: 145, SEQ ID NO: 149, SEQ ID NO: 153, SEQ ID NO: 154, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 162, SEQ ID NO: 166, SEQ ID NO: 170, SEQ ID NO: 174, SEQ ID NO: 178, SEQ ID NO: 182, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 191, SEQ ID NO: 195, SEQ ID NO: 199, SEQ ID NO: 203, SEQ ID NO:207 and SEQ ID NO: 211.

11. The pharmaceutical composition according to claim 1 , wherein said isolated mutated human or simian lentiviral ENV protein or said fragment of said isolated mutated human or simian lentiviral ENV protein comprises one of the amino acid sequences: SEQ ID NO: 96, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 99, SEQ ID NO: 100, SEQ ID NO: 104, SEQ ID NO: 125, SEQ ID NO: 126, SEQ ID NO: 127, SEQ ID NO: 128, SEQ ID NO: 129, SEQ ID NO: 133, SEQ ID NO: 154, SEQ ID NO: 155, SEQ ID NO: 156, SEQ ID NO: 157, SEQ ID NO: 158, SEQ ID NO: 162, SEQ ID NO: 183, SEQ ID NO: 184, SEQ ID NO: 185, SEQ ID NO: 186, SEQ ID NO: 187, SEQ ID NO: 191.

12. The pharmaceutical composition according to claim 1 , wherein said isolated mutated human or simian lentiviral ENV protein consists of one of the amino acid sequences: SEQ ID NO: 212, SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 220, SEQ ID NO: 224, SEQ ID NO: 228, SEQ ID NO: 232, SEQ ID NO: 236, SEQ ID NO: 240, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, SEQ ID NO: 244, SEQ ID NO: 245, SEQ ID NO: 249, SEQ ID NO: 253, SEQ ID NO: 257, SEQ ID NO: 261, SEQ ID NO: 265, SEQ ID NO: 269, SEQ ID NO: 420 and SEQ ID NO: 421.

13. The pharmaceutical composition according to claim 1 , wherein said isolated mutated human or simian lentiviral ENV protein consists of one of the amino acid sequences: SEQ ID NO: 212, SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 220, SEQ ID NO: 241, SEQ ID NO: 242, SEQ ID NO: 243, SEQ ID NO: 244, SEQ ID NO: 245, SEQ ID NO: 249, SEQ ID NO: 420 and SEQ ID NO: 421.

14. The pharmaceutical composition according to claim 1 , wherein said mutated protein consists of one of the amino acid sequences of the group consisting of SEQ ID NO: 271 to 283.

15. The pharmaceutical composition according to claim 1 , in association with at least one antiviral compound, preferably for a simultaneous, separated or sequential use.

16. The pharmaceutical composition according to claim 1 , for its use for stimulating an immune response in a host organism.

17. A method to obtain the active substance of a pharmaceutical composition, as defined in claim 1 , consisting of modifying the immunosuppressive property of:

a wild-type non-mutated human or simian lentiviral ENV protein,

or a fragment of said wild-type non-mutated human or simian lentiviral ENV protein, said fragment comprising at least 40 amino acids,

said wild-type non-mutated human or simian lentiviral ENV protein or fragment thereof presenting a transmembrane subunit (TM) comprising an immunosuppressive domain (ISU) containing the following amino acid sequence:

A-[I/V]-E-[K/R]-X′ a -X′ b -X-D-Q,

(SEQ ID NO: 427)

wherein

X represents any amino acid,

X′ a is C, D, E, H, I, K, M, N, P, Q, S, T, V, W or Y, and

X′ b is C, D, E, H, I, K, L, M, N, P, Q, S, T, V, W or Y,

said method comprising a step of introduction of at least one mutation of X′ a and/or X′ b r

to obtain:

an isolated mutated human or simian lentiviral ENV protein having no or a reduced immunosuppressive activity as compared to the wild type non-mutated human or simian lentiviral ENV protein,

or a fragment of said isolated mutated human or simian lentiviral ENV protein having no or a reduced immunosuppressive activity as compared to the wild type non-mutated human or simian lentiviral ENV protein,

said fragment comprising at least 40 amino acids,

said isolated mutated human or simian lentiviral ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q,

(SEQ ID NO: 416)

wherein

X represents any amino acid,

and either

X a is A, F, G, L or R, and X b is any amino acid,

X a is any amino acid, and X b is R, or

X a is A, F, G, L or R, and X b is R,

said substantial absence or reduction of immunosuppressive activity of the above mentioned mutated human or simian lentiviral ENV protein or of the above defined fragment being liable to be assessed by the fact that in an in vivo assay involving engrafted tumor cells rejection,

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 the corresponding wild type non-mutated 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).

18. A method to obtain the active substance of a pharmaceutical composition, as defined in claim 1 , consisting of modifying the immunosuppressive property of:

a wild-type non-mutated human or simian lentiviral ENV protein,

or a fragment of said wild-type non-mutated human or simian lentiviral ENV protein, said fragment comprising at least 40 amino acids,

A-[I/V]-E-[K/R]-Y-L-X-D-Q,

(SEQ ID NO: 1)

wherein

X represents any amino acid,

said method comprising a step of introduction of at least one mutation of Y in position 5 and/or L in position 6, to obtain:

an isolated mutated human or simian lentiviral ENV protein having no or a reduced immunosuppressive activity as compared to the wild type non-mutated human or simian lentiviral ENV protein,

or a fragment of said isolated mutated human or simian lentiviral ENV protein having no or a reduced immunosuppressive activity as compared to the wild type sequence, said fragment comprising at least 40 amino acids, said isolated mutated human or simian lentiviral ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q,

(SEQ ID NO: 416)

wherein

X represents any amino acid,

and either

X a is A, F, G, L or R, and X b is L, I, V, M or P,

X a is Y, I, H, C or T, and X b is R, or

X a is A, F, G, L or R, and X b is R.

19. A method to obtain the active substance of a pharmaceutical composition, as defined in claim 1 ,

wherein said isolated mutated human or simian lentiviral ENV protein having no or a reduced immunosuppressive activity as compared to the wild type non-mutated human or simian lentiviral ENV protein,

said fragment comprising at least 40 amino acids,

said isolated mutated human or simian lentiviral ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q,

(SEQ ID NO: 416)

wherein

X represents any amino acid,

and either

X a is A, F, G, L or R, and X b is L,

X a is Y, and X b is R, or

X a is A, F, G, L or R, and X b is R.

20. A method to obtain the active substance of a pharmaceutical composition, as defined in claim 1 ,

wherein said isolated mutated human or simian lentiviral ENV protein having no or a reduced immunosuppressive activity as compared to the wild type non-mutated human or simian lentiviral ENV protein,

said fragment comprising at least 40 amino acids,

said isolated mutated human or simian lentiviral ENV protein and fragment thereof comprising a mutated immunosuppressive domain (ISU) containing the following amino sequence:

A-[I/V]-E-[K/R]-X a -X b -X-D-Q,

(SEQ ID NO: 416)

wherein

X represents any amino acid,

and either

X a is A, F, G, L or R, and X b is L, or

X a is Y, and X b is R.

21. The pharmaceutical composition according to claim 1 , wherein said isolated mutated human lentiviral ENV protein or said fragment of said isolated mutated human lentiviral ENV protein has no or a reduced immunosuppressive activity as compared to a wild type non-mutated HIV-1 ENV protein consisting of the amino acid sequence SEQ ID NO: 417.

22. The pharmaceutical composition according to claim 1 , wherein said isolated mutated human lentiviral ENV protein or said fragment of said isolated mutated human lentiviral ENV protein has no or a reduced immunosuppressive activity as compared to a wild type non-mutated HIV-2 ENV protein consisting of the amino acid sequence SEQ ID NO: 418.

23. The pharmaceutical composition according to claim 1 , wherein said isolated mutated simian lentiviral ENV protein or said fragment of said isolated mutated simian lentiviral ENV protein has no or a reduced immunosuppressive activity as compared to a wild type non-mutated SIV ENV protein consisting of the amino acid sequence SEQ ID NO: 419.

24. The pharmaceutical composition according to claim 1 , wherein said isolated mutated human lentiviral ENV protein or said fragment of said isolated mutated human lentiviral ENV protein has no or a reduced immunosuppressive activity as compared to a HIV-1 ENV protein comprising a wild type non-mutated immunosuppressive domain selected from the group consisting of: SEQ ID NO: 366, SEQ ID NO: 367, SEQ ID NO: 368, SEQ ID NO: 369, SEQ ID NO: 370 and SEQ ID NO: 371.

25. The pharmaceutical composition according to claim 1 , wherein said isolated mutated human lentiviral ENV protein or said fragment of said isolated mutated human lentiviral ENV protein has no or a reduced immunosuppressive activity as compared to a HIV-2 ENV protein comprising a wild type non-mutated immunosuppressive domain selected from the group consisting of: SEQ ID NO: 376 and SEQ ID NO: 377.

26. The pharmaceutical composition according to claim 1 , wherein said isolated mutated simian lentiviral ENV protein or said fragment of said isolated mutated simian lentiviral ENV protein has no or a reduced immunosuppressive activity as compared to a SIV ENV protein comprising a wild type non-mutated immunosuppressive domain selected from the group consisting of: SEQ ID NO: 372, SEQ ID NO: 373, SEQ ID NO: 374 and SEQ ID NO: 375.

Assignments (2)

MERGER Recorded Oct 14, 2021

From: UNIVERSITE PARIS-SUD XI

To: UNIVERSITE PARIS -SACLAY

Reel/Frame 057794/0920 →

ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 4, 2013

From: HEIDMANN, THIERRY

To: CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUE; UNIVERSITE PARIS-SUD XI; INSTITUT GUSTAVE ROUSSY; VIROXIS S.A.S.

Reel/Frame 029745/0258 →

Continuity (2)

Provisional Application 61567810 · Dec 7, 2011

Related Publication 20130149323A1 · Jun 13, 2013