Methods and compositions for inducing an immune response to EGFRvIII

• US 9,200,057 B2
• Filed: 11/17/2011
• Issued: 12/01/2015
• Est. Priority Date: 11/17/2010
• Status: Active Grant

• Current Assignee: Aduro Biotech, Inc. (Novartis Ag), Providence Health & Services - Oregon (Providence St. Joseph Health)
• Original Assignee: Aduro Biotech, Inc. (Novartis Ag), Providence Health & Services - Oregon (Providence St. Joseph Health)
• Inventors: Lauer, Peter M., Bahjat, Keith
• Primary Examiner(s): NAVARRO, ALBERT MARK
• Application Number:
  US13/988,076
• Publication Number:
  US 20140037662A1
• Time in Patent Office: 1,475 Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/522 avirulent or attenuated
• CPC Class Codes: A61K 2039/523 expressing foreign proteins
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 39/001104 Epidermal growth factor receptors [EGFR]
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: C07K 14/71 for growth factors; for growth regulators

Abstract

The present invention relates to methods of inducing a T-cell response against a EGFRvIII in a subject. These method comprise administering to a subject a composition which expresses at least one immunogenic polypeptide, the amino acid sequence of which comprise a plurality of EGFRvIII polypeptide sequences, the sequence of which each comprise EEKKGNYV (SEQ ID NO: 3), and/or administering the polypeptide itself.

Claims

1. A composition comprising:

a bacterium or virus which comprises a nucleic acid encoding at least one immunogenic polypeptide, the amino acid sequence of which comprise a plurality of EGFRvIII polypeptide sequences, the sequence of which each comprise EEKKGNYV (SEQ ID NO;

     3), and wherein said plurality of EGFRvIII polypeptide sequences comprise at least three copies of PASRALEEKKGNYVVTDHGSC (SEQ ID NO;

     5).

2. The composition of claim 1 wherein said plurality of EGFRvIII polypeptide sequences comprise at least five copies of PASRALEEKKGNYVVTDHGSC (SEQ ID NO:

5).

3. The composition of claim 1, wherein each EGFRvIII polypeptide sequence is flanked by a sequence which is configured to be cleaved by the proteasome.

4. A pharmaceutical composition comprising:

the composition of claim 1; and

a pharmaceutically acceptable excipient.

5. The composition of claim 1 wherein composition comprises an attenuated Listeria monocytogenes bacterium comprising said nucleic acid.

6. The composition of claim 5, wherein the composition comprises a Listeria monocytogenes bacterium comprising said nucleic acid integrated into the genome of said bacterium.

7. The composition of claim 5, wherein the bacterium is an actA deletion mutant or an actA insertion mutant, an inlB deletion mutant or an inlB insertion mutant or a Δ

actA/Δ

inlB mutant comprising both an actA deletion or an actA insertion and an inlB deletion or an inlB insertion.

8. The composition of claim 5, wherein said nucleic acid has been integrated into a virulence gene of said bacterium, and the integration of the polynucleotide disrupts expression of the virulence gene or disrupts a coding sequence of the virulence gene.

9. The composition of claim 8, wherein the virulence gene is actA or inlB.

10. The composition of claim 9, wherein the bacterium is Lm Δ

actA/Δ

inlB.

11. The composition of claim 7, wherein the bacterium further comprises a genetic mutation that attenuates the ability of the bacterium to repair nucleic acid.

12. The composition of claim 11, wherein the genetic mutation is in one or more genes selected from phrB, uvrA, uvrB, uvrC, uvrD and recA.

13. The composition of claim 9, wherein the bacterium is a Listeria monocytogenes prfA mutant, the genome of which encodes a prfA protein which is constitutively active.

14. The composition of claim 10, wherein the bacterium is a killed but metabolically active Listeria monocytogenes.

15. The composition of claim 5, wherein the bacterium is a Listeria monocytogenes prfA mutant, the genome of which encodes a prfA protein which is constitutively active.

16. The composition of claim 5, wherein the nucleic acid is codon optimized for expression by Listeria monocytogenes.

17. The composition of claim 5, wherein said composition further comprises a pharmaceutically acceptable excipient.

18. The composition of any one of claims 5, wherein said nucleic acid encodes said immunogenic polypeptide as a fusion protein comprising a secretory signal sequence.

19. The composition of claim 18, wherein the secretory signal sequence is a Listeria monocytogenes ActA signal sequence.

20. The composition of claim 19, wherein said nucleic acid molecule encodes said immunogenic polypeptide as a fusion protein comprising an in frame ActA-N100 sequence selected from the group consisting of selected from the group consisting of SEQ ID NO:

28, SEQ ID NO;

29, SEQ ID NO;

30, and SEQ ID NO;

31, or an amino acid sequence having at least 90% sequence identity to said ActA-N100 sequence.

21. The composition of claim 5, wherein said nucleic acid encodes said immunogenic polypeptide as a fusion protein, wherein:

each of the plurality of EGFRvIII polypeptide sequences has the sequence PASRALEEKKGNYVVTDHGSC (SEQ ID NO;

     5);

each of the plurality of EGFRvIII polypeptide sequences is flanked by a sequence which is configured to be cleaved by the proteasome;

said fusion protein further comprises an ActA-N100 sequence selected from the group consisting of SEQ ID NO;

28, SEQ ID NO;

29, SEQ ID NO;

30, and SEQ ID NO;

31 or an amino acid sequence having at least 90% sequence identity to said ActA-N100 sequence; and

said fusion protein is expressed from a nucleic acid sequence operably linked to a Listeria monocytogenes ActA promoter.