Aβ immunogenic peptide carrier conjugates and methods of producing same

• US 9,095,536 B2
• Filed: 08/20/2007
• Issued: 08/04/2015
• Est. Priority Date: 12/17/2003
• Status: Expired due to Fees

• Current Assignee: Janssen Sciences Ireland UC (Johnson & Johnson), Wyeth LLC (Pfizer Inc.)
• 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:
  US11/841,919
• Publication Number:
  US 20080145373A1
• Time in Patent Office: 2,906 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. A method of producing an immunogenic conjugate, the method comprising the steps of:

(a) adding a carboxy-terminal cysteine residue to an Aβ

fragment comprising residues 1-7 of Aβ

(SEQ ID NO;

21);

(b) derivatizing one or more lysine residues of a CRM₁₉₇ carrier protein to generate an activated functional group on the carrier protein;

(c) reacting the peptide immunogen comprising residues 1-7 of Aβ

(SEQ ID NO;

21) and the carboxy-terminal cysteine residue with the carrier protein of step (b) under conditions to form a conjugate; and

(d) further reacting the conjugate of step (c) with N-acetylcysteamine to inactivate any remaining activated functional group on the carrier protein to generate the immunogenic conjugate,

wherein the immunogenic conjugate has the formula;

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

3. The method of claim 2, wherein n is an integer greater than or equal to 12, but less than or equal to 15.

4. The method of claim 3, wherein n is 12.

5. The method of claim 3, wherein n is 14.

6. The method of claim 3, wherein the peptide immunogen is DAEFRHD-C (SEQ ID NO:

2).

7. The method of claim 1, wherein the immunogenic conjugate has the formula:

8. The method of claim 7, wherein n is 12, 14 or 15.

9. The method of claim 8, wherein n plus p equals from 20 to 22.

10. The method of claim 7, wherein n plus p equals from 20 to 22.