A-β immunogenic peptide carrier conjugates and methods of producing same

• US 9,125,847 B2
• Filed: 02/14/2012
• Issued: 09/08/2015
• Est. Priority Date: 12/17/2003
• Status: Expired due to Fees

• Current Assignee: Janssen Sciences Ireland UC (Johnson & Johnson)
• Original Assignee: Janssen Sciences Ireland UC (Johnson & Johnson), Wyeth LLC (Pfizer Inc.)
• Inventors: Arumugham, Rasappa G., Prasad, A. Krishna, Hagen, Michael
• Primary Examiner(s): GUDIBANDE, SATYANARAYAN R
• Application Number:
  US13/396,543
• Publication Number:
  US 20120207706A1
• Time in Patent Office: 1,302 Days
• Field of Search: None
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/6031 Proteins
• CPC Class Codes: A61K 2039/6081 Albumin; Keyhole limpet haemocyanin [KLH]
• CPC Class Codes: A61K 2039/62 characterised by the link between antigen and carrier
• CPC Class Codes: A61K 39/0007 Nervous system antigens; Prions
• CPC Class Codes: A61K 47/643 Albumins, e.g. HSA, BSA, ovalbumin or a Keyhole Limpet Hemocyanin [KHL]
• CPC Class Codes: A61K 47/646 the entire peptide or protein drug conjugate elicits an immune response, e.g. conjugate vaccines
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 31/12 Antivirals
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/04 Immunostimulants
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: Y02A 50/30 Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Abstract

The present invention is directed to methods of producing conjugates of Aβ peptide immunogens with protein/polypeptide carrier molecules, which are useful as immunogens, wherein peptide immunogens are conjugated to protein carriers via activated functional groups on amino acid residues of the carrier or of the optionally attached linker molecule, and wherein any unconjugated reactive functional groups on amino acid residues are inactivated via capping, thus retaining the immunological functionality of the carrier molecule, but reducing the propensity for undesirable reactions that could render the conjugate less safe or effective. Furthermore, the invention also relates to such immunogenic products and immunogenic compositions containing such immunogenic products made by such methods.

Claims

1. An immunogenic conjugate having the formula:

2. The immunogenic conjugate of claim 1, wherein the peptide immunogen further comprises a terminal cysteine residue and the peptide immunogen is covalently attached to the derivatized functional group of the amino acid residue of the carrier protein via the terminal cysteine residue.

3. The immunogenic conjugate of claim 2, wherein the terminal cysteine residue is located at the carboxy-terminus of the Aβ

fragment.

4. The immunogenic conjugate of claim 2, wherein the terminal cysteine residue is located at the amino-terminus of the Aβ

fragment.

5. The immunogenic conjugate of claim 1, wherein the capping molecule is selected from the group consisting of an amino group, a hydroxyl group, a cysteamino group, an N-acetylcysteamino group, and an ethanolamino group.

6. The immunogenic conjugate of claim 5, wherein the capping molecule is an N-acetylcysteamino group.

7. The immunogenic conjugate of claim 1, wherein n is an integer greater than or equal to 5, but less than or equal to 25.

8. The immunogenic conjugate of claim 7, wherein n is an integer greater than or equal to 12, but less than or equal to 20.

9. The immunogenic conjugate of claim 8, wherein n is an integer greater than or equal to 12, but less than or equal to 15.

19. An immunogenic composition, comprising an immunogenic conjugate of claim 1, together with one or more pharmaceutically acceptable excipients, diluents, and/or adjuvants.

20. The immunogenic composition of claim 19, wherein one or more adjuvants are selected from the group consisting of GM-CSF, 529 SE, IL-12, aluminum phosphate, aluminum hydroxide, Mycobacterium tuberculosis, Bordetella pertussis, bacterial lipopolysaccharides, aminoalkyl glucosamine phosphate compounds, MPL™

(3-O-deacylated monophosphoryl lipid A), a polypeptide, Quil A, STIMULON™

QS-21, a pertussis toxin (PT), an E. coli heat-labile toxin (LT), IL-1α

, IL-1β

, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-13, IL-14, IL-15, IL-16, IL-17, IL-18, interferon-α

, interferon-β

, interferon-γ

, G-CSF, TNF-α and

TNF-β

.

10. An immunogenic conjugate having the formula:

11. The immunogenic conjugate of claim 10, wherein n is an integer greater than or equal to 5, but less than or equal to 25.

12. The immunogenic conjugate of claim 11, wherein n is an integer greater than or equal to 12, but less than or equal to 20.

13. The immunogenic conjugate of claim 12, wherein n is an integer greater than or equal to 12, but less than or equal to 15.

14. The immunogenic conjugate of claim 13, wherein n is 12, 14 or 15.

15. The immunogenic conjugate of claim 10, wherein the peptide immunogen is KLVFFAED-C (SEQ ID NO:

45).

16. The immunogenic conjugate of claim 10, wherein the immunogenic conjugate has the formula:

17. The immunogenic conjugate of claim 16, wherein n plus p equals from 20 to 22.

18. The immunogenic conjugate of claim 16, wherein n is 12, 14 or 15.