WO2012135616A1

WO2012135616A1 - Vaccination against advanced glycation end-products

Info

Publication number: WO2012135616A1
Application number: PCT/US2012/031446
Authority: WO
Inventors: Lewis Gruber
Original assignee: Siwa Corporation
Priority date: 2011-03-31
Filing date: 2012-03-30
Publication date: 2012-10-04

Abstract

A vaccine composition, for immunizing a human patient, comprises an AGE antigen, an adjuvant, a preservative, and optionally an excipient. A method of immunizing a human patient against AGE-modified proteins comprises administering the vaccine composition, in an amount effective to cause immunization against AGE- modified proteins. Immunizing a human patient against AGE-modified proteins may be used to treat atherosclerosis.

Description

VACCINATION AGAINST ADVANCED GLYCATION END-PRODUCTS

CROSS REFERENCE TO RELATED APPLICATION

[01] This application claims the benefit of provisional application no. 61/470,167 entitled "VACCINATION AGAINST ADVANCED GLYCATION END-PRODUCTS" filed 31 March 2011 , attorney docket no. SIW01-004-PRO, the entire contents of which are hereby incorporated by reference, except where inconsistent with the present application.

BACKGROUND

[02] Advanced glycation end-products (AGEs; also referred to AGE-modified

proteins) arise from a non-enzymatic reaction of sugars with protein side-chains in aging cells[1]. Hyperglycemia, caused by diabetes mellitus (DM), and oxidative stress promote this post-translational modification of membrane proteins[2]. AGEs have been associated with several pathological conditions including diabetic complications, inflammation, retinopathy, nephropathy, atherosclerosis, stroke, endothelial cell dysfunction, and neurodegenerative disorders[3].

[03] AGE-modified erythrocytes have been implicated in the pathogenesis of atherosclerosis, while the absence of AGE-modified erythrocytes has been correlated with reduced atherosclerosis[4]. Localization of AGEs in atherosclerotic lesions of the aorta in non-diabetic patients has been reported in intima cells, endothelial cells, smooth muscle cells, macrophages and foam cells[5]. The damage caused by AGE-modified cells may also lead to nephropathy, retinopathy, neuropathy, heart disease, stroke, and peripheral vascular disease[6].

[04] Vaccines have been widely used, since their introduction by Edward Jenner in the 1770s, to confer immunity against a wide range of diseases, starting from viruses and more recently to other afflictions, like cancer[7]. Vaccine preparations contain a selected immunogenic agent, such as an antigen (an agent that is the target of an immune response), capable of stimulating immunity to the antigen. Typical antigens utilized in vaccines include, for example, viruses, either killed or attenuated, purified viral components, and DNA[8]. Antigens used in the production of cancer vaccines include, for example, tumor-associated carbohydrate antigens (TACAs), DNA, dendritic cells, whole cells, and viral vectors[9]. Different techniques are employed to produce the desired amount and type of antigen being sought. For example, pathogenic viruses are grown either in eggs or cells[8]. Recombinant DNA technology is often utilized to generate attenuated vaccines[8].

[05] Immunity is a long-term immune response, either cellular or humoral[7]. A cellular immune response is activated when an antigen is presented preferably with a co-stimulator to a T-cell, which causes it to differentiate and produces

cytokines[10]. The cells involved in the generation of the cellular immune response are two classes of T-helper (Th) cells, Th1 and Th2[8]. Th1 cells stimulate B cells to produce predominantly antibodies of the lgG2A isotype, which activates the complement cascade and binds the Fc receptors of macrophages, while Th2 cells stimulate B cells to produce lgG1 isotype antibodies in mice, lgG4 isotype antibodies in humans, and IgE isotype antibodies[8]. Professional antigen-presenting cells include dendritic cells, macrophages, and B cells[8].

[06] A humoral immune response is triggered when a B cell selectively binds to an antigen and begins to proliferate, leading to the production of a clonal population of cells that produce antibodies that specifically recognize that antigen and which may differentiate into antibody-secreting cells, referred to as plasma-cells or memory-B cells[11]. Antibodies or immunoglobulins, are molecules produced by B-cells that bind the specific antigen[8]. The antigen-antibody complex also triggers several responses, either cell-mediated, for example by natural killers (NK) or macrophages, or serum-mediated, for example by activating the complement system, a complex of several serum proteins that act sequentially in a cascade that result in the lysis of the target cell[8].

[07] Immunological adjuvants (also referred to simply as "adjuvants") are the

component(s) of a vaccine which augment the immune response to the

immunogenic agent. Adjuvants function by attracting macrophages to the immunogenic agent and then presenting the agent to the regional lymph nodes to initiate an effective antigenic response. Adjuvants may also act as carriers themselves for the immunogenic agent. Adjuvants may induce an inflammatory response, which may play an important role in initiating the immune response[8]. Adjuvants include: mineral compounds such as aluminum salts, oil emulsions, bacterial products, liposomes, immunostimulating complexes, and squalene.

[08] Other components of vaccines include pharmaceutically acceptable

excipients, preservatives, diluents, and pH adjusters. A variety of these components of vaccines, as well as adjuvants, are described in

www.cdc.gov/vaccines/pubs/pinkbook/downloads/appendices/b/excipient-table- 1.pdf, www.cdc.gov/vaccines/pubs/pinkbook/downloads/appendices/b/excipient- table-2.pdf, and Vogel FR, er a/[15].

SUMMARY

[09] In a first aspect, the present invention is a vaccine composition, for

immunizing a human patient, comprising (a) an AGE antigen, (b) an adjuvant, (c) a preservative, and (d) optionally, an excipient.

[10] In a second aspect, the present invention is a method of treating

atherosclerosis, comprising immunizing a human patient in need thereof against AGE-modified proteins.

[1 1] In a third aspect, the present invention is a method of immunizing a human patient against AGE-modified proteins, comprising administering a vaccine composition, in an amount effective to cause immunization against AGE-modified proteins.

DETAILED DESCRIPTION

[12] The present invention makes use of the discovery that enhanced clearance of cells containing AGE-modified proteins (AGE-modified cells), such as erythrocytes, is beneficial in reducing cardiovascular disease, especially when present as a complication of diabetes, or the pre-diabetic condition referred to as "Syndrome-X". Elevated blood glucose concentrations lead to modifications of protein side chains in cells, including circulating erythrocytes and other cell types. Non-enzymatic glycation of membrane proteins results in the formation of AGE-modified cells, which cause reduced cell deformability that is associated with the formation of atherosclerotic lesions. Vaccination against AGE-modified proteins produces the desired result of controlling the production of AGE-modified proteins in a subject in need thereof. The continuous and virtually ubiquitous surveillance exercised by the immune system in the body in response to a vaccination, allows maintaining low levels of AGE-modified cells in the body. In particular, vaccination against AGE-modified proteins, particularly AGE-modified proteins of erythrocytes, may be used to treat

atherosclerosis.

AGE-modified proteins are also a marker of partially- and non-functional cells. Vaccination against AGE-modified proteins can also help remove or kill these partially- and non-functional cells, to enhance regeneration by allowing functional cells to replace them. See, for example, WO 2009/1 341 1 to Gruber (26 Nov.

2009). A vaccine against AGE-modified proteins can be used in a method for promoting tissue or organ regeneration in a subject by killing partially-functional or non-functional cells. The vaccine can also have cosmetic effects, for example reducing aging or the appearance of aging, of the skin and/or hair. These methods are particularly applicable to diabetic persons, where high blood glucose levels enhance the presence of AGE-modified proteins.

Vaccines against AGE-modified proteins contain an AGE antigen, an adjuvant, a preservative and optional excipients. AGE antigens are AGE-modified proteins, and mixtures thereof, which can elicit an immune response in humans, preferably when administered in the presence of one or more adjuvants. Examples include AGE-antithrombin III, AGE-calmodulin, AGE-insulin, AGE-ceruloplasmin, AGE-collagen, AGE-cathepsin B, AGE-albumin, AGE-crystallin, AGE-plasminogen activator, AGE-endothelial plasma membrane protein, AGE-aldehyde reductase, AGE-transferrin, AGE-fibrin, AGE-copper/zinc SOD, AGE-apo B, AGE-fibronectin, AGE-pancreatic ribose, AGE-apo A-l and II, AGE-hemoglobin, AGE-Na⁺/K⁺-ATPase, AGE-plasminogen, AGE-myelin, AGE-lysozyme, AGE-immunoglobulin, AGE-red cell Glu transport protein, ΑΘΕ-β-Ν-acetyl hexominase, AGE-apo E, AGE-red cell membrane protein, AGE-aldose reductase, AGE-ferritin, AGE-red cell spectrin, AGE- alcohol dehydrogenase, AGE-haptoglobin, AGE-tubulin, AGE-thyroid hormone, AGE-fibrinogen, AGE- ₂-microglobulin, AGE-sorbitol dehydrogenase, AGE-c - antitrypsin, AGE-carbonate dehydratase, AGE-RNAse, AGE-low density lipoprotein, AGE-hexokinase and AGE-apo C-l. These proteins may be produced by isolation of the protein from natural sources or recombinantly, followed by AGE modification, for example by incubation in a 0.1-3 M solution of glucose, glucose-6-phosphate, fructose or ribose, for 10-100 days, where the solution is buffered with a phosphate buffer at a pH of 6-8. Fragments and digestates of the proteins may also be used, followed by AGE modification, as long as the fragments and digestates contain reducing amino acids, so that a Schiff base is formed and the Amadori reaction can take place to form the corresponding ketoamine. AGE-modified cells, such as AGE- modified erythrocytes, whole, lysed, or partially digested, may also be used. Further details of some of these AGE-modified proteins and their preparation are described in U.S. Patent No. 5,702,704 to Bucala (issued December 30, 1997) (hereby incorporated by reference).

Preferably, the AGE-modified proteins which are targets of the vaccine are AGE-modified proteins of cell, and exclude non-cellular modified proteins, for example exclude AGE-modified albumin and/or AGE-modified collagen. Preferably, the AGE antigen is a condensation product advanced glycation end-product comprising a lysine component, an argine component and a reducing sugar component, as described in U.S. Patent No. 6,380,165 to Al-Abed et al. (issued April 30, 2002) (hereby incorporated by reference). Examples include an advanced glycation end-product comprising a condensation product of a lysyl component, an arginyl component and a reducing sugar component, wherein the arginyl component and the reducing sugar component form a monocyclic substituted imidazole; for example a compound according to formula I:

wherein a lysyl component is indicated by the box labeled "K"; an arginyl component is indicated by the boxy labeled "R"; and a reducing sugar component is not boxed; and wherein Ri and R₄ are independently H or an amide bond to an amino acid residue or a peptide chain; R₂ and R₃ are, independently, OH or an amide bond to an amino acid residue or a peptide chain; R₅ is H, CH₂OH or CHOHCH₂OH; and wherein if more than one of R-i , R₂, R₃ or R₄ is an amide bond, then the lysyl "K" component and the arginyl "R" component may be amino acid residues of the same or a different peptide chain.

Other preferred AGE antigens are AGE-RNAse, AGE-hemoglobin such as AGE-human hemoglobin, AGE-albumin such as AGE-bovine serum albumin and AGE-human serum albumin, AGE-low density lipoprotein (AGE-LDL) and AGE- collagen IV, such as those described in U.S. Patent No. 5,702,704 to Bucala (issued December 30, 1997).

Preferably, the vaccine used to vaccinate a human contains from 1 microgram to 100 milligrams of at least one AGE antigen, including 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 400, 800 or 1000 micrograms, or 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80 or 90 milligrams. The amount used for a single injection corresponds to a unit dosage.

Adjuvants included: mineral compounds such as aluminum salts, oil emulsions, bacterial products, liposomes, immunostimulating complexes, and squalene. Aluminum compounds are the most widely used adjuvants in human and veterinary vaccines. These aluminum compounds include aluminum salts such as aluminum phosphate (AIP0₄) and aluminum hydroxide (AI(OH)₃) compounds, typically in the form of gels, and are generically referred to in the field of vaccine immunological adjuvants as "alum". Aluminum hydroxide is a poorly crystalline aluminum oxyhydroxide having the structure of the mineral boehmite. Aluminum phosphate is an amorphous aluminum hydroxyphosphate. Negatively charged species (for example, negatively charged antigens) can absorb onto aluminum hydroxide gels at neutral pH, whereas positively charged species (for example, positively charged antigens) can absorb onto aluminum phosphate gels at neutral pH. It is believed that these aluminum compounds provide a depot of antigen at the site of administration (for example, injection), thereby providing a gradual and continuous release of antigen to stimulate antibody production. Aluminum compounds tend to more effectively stimulate a cellular response mediated by Th2, rather than Th1 cells[12].

[19] Emulsion adjuvants include water-in-oil emulsions (for example, Freund's adjuvants, such as killed mycobacteria in oil emulsion) and oil-in-water emulsions (for example, F-59). Emulsion adjuvants include an immunogenic component, for example squalene (MF-59) or mannide oleate (Incomplete Freund's Adjuvants), which can induce an elevated humoral response, increased T cell proliferation, cytotoxic lymphocytes, and cell-mediated immunity.

[20] Liposomal or vesicular adjuvants (including paucilamellar lipid vesicles) have lipophilic bilayer domains and an aqueous milieu which can be used to encapsulate and transport a variety of materials, for example an antigen. Paucilamellar vesicles (for example, those described in U.S. Pat. No. 6,387,373) can be prepared by mixing, under high pressure or shear conditions, a lipid phase comprising a non- phospholipid material (for example, an amphiphile surfactant; see U.S. Pat. Nos. 4,217,344; 4,917,951 ; and 4,911 ,928), optionally a sterol, and any water-immiscible oily material to be encapsulated in the vesicles (for example, an oil such as squalene oil and an oil-soluble or oil-suspended antigen); and an aqueous phase such as water, saline, buffer or any other aqueous solution used to hydrate the lipids.

Liposomal or vesicular adjuvants are believed to promote contact of the antigen with immune cells, for example by fusion of the vesicle to the immune cell membrane, and preferentially stimulate the Th1 sub-population of T-helper cells.

[21] Other types of adjuvants include Mycobacterium bovis bacillus Calmette-

Guerin (BCG), quill-saponin, unmethylated CpG dinucleotides (CpG motifs)[13]. Additional adjuvants are described in U.S. Patent Application Publication, Pub. No. US 2010/0226932 (Sept. 9, 2010), and Jiang, ZH er a/[14]. Preferable adjuvants include Freund's complete adjuvant and Freund's incomplete adjuvant.

[22] One or more preservatives, such as antioxidants, antibacterial and

antimicrobial agents, as well as combinations thereof, are also included in the vaccine. Examples include benzethonium chloride, ethylenediamine-tetraacetic acid sodium (EDTA), thimerosal, phenol, 2-phenoxyethanol, formaldehyde and formalin; antibacterial agents such as amphotericin B, chlortetracycline, gentamicin, neomycin, polymyxin B and streptomycin; antimicrobial surfactants such as polyoxyethylene-9, 10-nonyl phenol (Triton N-101 , octoxynol-9), sodium

deoxycholate and polyoxyethylated octyl phenol (Triton X-I00).

[23] Other components of vaccines include pharmaceutically acceptable

excipients, such as stabilizers, thickening agents, toxin detoxifiers, diluents, pH adjusters, tonicity adjustors, surfactants, antifoaming agents, protein stabilizers, dyes and solvents. Examples of such excipients include hydrochloric acid, phosphate buffers, sodium acetate, sodium bicarbonate, sodium borate, sodium citrate, sodium hydroxide, potassium chloride, potassium chloride, sodium chloride,

polydimethylsilozone, brilliant green, phenol red (phenolsulfon-phthalein), glycine, glycerin, sorbitol, histidine, monosodium glutamate, potassium glutamate, sucrose, urea, lactose, gelatin, sorbitol, polysorbate 20, polysorbate 80 and glutaraldehyde. A variety of these components of vaccines, as well as adjuvants, are described in www.cdc.gov/vaccines/pubs/pinkbook/downloads/appendices/b/excipient-table- 1.pdf, www.cdc.gov/vaccines/pubs/pinkbook/downloads/appendices/b/excipient- table-2.pdf, and Vogel FR, et a/[15]. Preferably, the vaccine is provided in unit dosage form, or multidosage form, such as 2-100, or 2-10 doses in a packed, sterilized, ready-to-inject form. The unit dosages may be provided in a vial with a septum, or in a syringe with or without a needle. The vaccine may be administered intravenously, subdermally, or intraperitoneally. For vaccination, preferably the vaccine may be administered one or more times, such as 1 to 10 times, including 2, 3, 4, 5, 6, 7, 8 or 9 times, over a period of time ranging from 1 week to 1 year, 2-10 weeks or 2-10 months.

Furthermore, booster vaccinations may be desirable, over the course of 1 year to 20 years, including 2, 5, 10 and 15 years.

EXAMPLES

Example 1 (Prophetic): AGE-RNAse containing vaccine in a human subject.

AGE-RNAse is prepared by incubating RNAse in a phosphate buffer solution containing 0.1 -3M glucose, glucose-6-phosphate, fructose, or ribose for 10-100 days. The AGE-RNAse solution is dialyzed and the protein content is measured. Virella et a/.[25]. Aluminum hydroxide or aluminum phosphate, as an adjuvant, is added to 100 microgram of the AGE-RNAse. Formaldehyde or formalin is added as a preservative to the preparation. Moreover, ascorbic acid is added as an antioxidant. The vaccine also includes other excipients: phosphate buffer to adjust the pH, and glycine as a protein stabilizer. The composition is injected into a human subject subcutaneously and the titer of AGE-RNAse antibody is determined by ELISA one month after the inoculation. Ameli et al. [26].

Example 2 (Prophetic): Injection regimen for an AGE-RNAse containing vaccine in a human subject.

The same vaccine as in Example 1 is injected into a human subject. The titer of AGE-RNAse is determined by ELISA after two weeks. Additional injections are performed after three weeks and six weeks, respectively. Further titer determination is performed two weeks after each injection. Example 3 (Prophetic): An AGE-hemoglobin containing vaccine in a human subject.

AGE-hemoglobin is prepared by incubating human hemoglobin in a phosphate buffer solution containing 0.1-3M glucose, glucose-6-phosphate, fructose, or ribose for 10-100 days. The AGE-hemoglobin solution is dialyzed and the protein content is measured. All vaccine components are the same as in Example 1 , except AGE-hemoglobin is substituted for AGE-RNAse. Administration is carried out as in Example 1 , or as in Example 2.

Example 4 (Prophetic): AGE-human serum albumin containing vaccine in a human subject.

AGE-human serum albumin is prepared by incubating human serum albumin in a phosphate buffer solution containing 0.1-3M glucose, glucose-6-phosphate, fructose, or ribose for 10-100 days. The AGE-human serum albumin solution is dialyzed and the protein content is measured. All vaccine components are the same as in Example 1 , except AGE-human serum albumin is substituted for AGE-RNAse. Administration is carried out as in Example 1 , or as in Example 2.

[34] REFERENCES

[35] 1. Ando K, et al., "Membrane Proteins of Human Erythrocytes Are Modified by

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[36] 2. Lindsey JB, et al,, "Receptor For Advanced Glycation End-Products

(RAGE) and soluble RAGE (sRAGE): Cardiovascular Implications," Diabetes Vascular Disease Research, Vol. 6(1), 7-14, (2009).

[37] 3. Bierhaus A, "AGEs and their interaction with AGE-receptors in vascular disease and diabetes mellitus. I. The AGE concept," Cardiovasc Res, Vol. 37(3), 586-600 (1998).

[38] 4. Jandeleit-Dahm K, et al., "The AGE/RAGE Axis in Diabetes-Accelerated

Atherosclerosis," Clinical and Experimental Pharmacology and Physiology, Vol. 35, 329-334 (2008).

[39] 5. Sakata N. et al., "Immunohistochemical Localization of Different Epitopes of

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[40] 6. Karachalias N. et al., "Accumulation of Fructosyl-Lysine and Advanced

Glycation End Products in the Kidney, Retina and Peripheral Nerve of

Streptozotocin-lnduced Diabetic Rats." Biochemical Society Transactions, 31 , 1423- 25 (2003).

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804-12 (2007).

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Differentiation," Trends Immunol, Vol. 30(6), 277-85 (2009).

[46] 12. Coler R et al. , "Development and Characterization of Synthetic

Glucopyranosyl Lipid Adjuvant System as a Vaccine Adjuvant," PLoS ONE, Vol. 6(1): e16333 (201 1 ).

[47] 13. Cheadle EJ, "Bugs as Drugs for Cancer," Immunology, Vol. 107, 10-19

(2002).

[48] 14. Jiang, ZH et al. "Synthetic vaccines: the role of adjuvants in immune targeting," Curr Med Chem 10(15): 1423-39 (2003).

[49] 15. Vogel FR, et al. "A compendium of vaccine adjuvants and excipients,"

Pharm Biotechnol 6: 141-228 (1995).

[50] 16. U.S. Patent No. 5,702,704 to Bucala (issued December 30, 1997).

[51] 17. U.S. Patent No. 6,380, 165 to Al-Abed et al. (issued April 30, 2002).

[52] 18. U.S. Patent No. 6,387,373 to Wright et al. (issued May 14, 2002).

[53] 19. U.S. Patent No. 4,217,344 to Vanlerberghe et al. (issued August 12,

1980).

[54] 20. U.S. Patent No. 4,917,951 to Wallach (issued April 17, 1990).

[55] 21. U.S. Patent No. 4,911 ,928 to Wallach (issued March 27, 1990).

[56] 22. U.S. Application Publication, Pub. No. US 2010/226932 to Smith et al.

(September 9, 2010). [57] 23. www.cdc.gov/vaccines/pubs/pinkbook/downloads/appendices/b/ excipient-table-1.pdf (The Pink Book, Epidemiology and Prevention of Vaccine Preventable Diseases, 1 1^th Ed., May 2009).

[58] 24. www.cdc.gov/vaccines/pubs/pinkbook/downloads/appendices/b/

excipient-table-2.pdf (The Pink Book, Epidemiology and Prevention of Vaccine Preventable Diseases, 1 1^th Ed., May 2009).

[59] 25. Virella G et al. , "Autoimmune Response to Advanced Glycosylation End-

Products of Human LDL," Journal of Lipid Res, Vol. 44, 487-493 (2003).

[60] 26. Ameli S et al. , "Effect of Immunization With Homologous LDL and

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Claims

What is claimed is:

1. A vaccine composition, for immunizing a human patient, comprising:

(a) an AGE antigen,

(b) an adjuvant,

(c) a preservative, and

(d) optionally, an excipient.

2. The vaccine composition of claim 1 , wherein the AGE antigen is an AGE-modified protein selected from the group consisting of AGE-antithrombin III, AGE-calmodulin, AGE-insulin, AGE-ceruloplasmin, AGE-collagen, AGE-cathepsin B, AGE-albumin, AGE-crystallin, AGE-plasminogen activator, AGE-endothelial plasma membrane protein, AGE-aldehyde reductase, AGE-transferrin, AGE-fibrin, AGE- copper/zinc SOD, AGE-apo B, AGE-fibronectin, AGE-pancreatic ribose, AGE-apo A- I and II, AGE-hemoglobin, AGE-Na7K⁺-ATPase, AGE-plasminogen, AGE-myelin, AGE-lysozyme, AGE-immunoglobulin, AGE-red cell Glu transport protein, AGE-β-Ν- acetyl hexominase, AGE-apo E, AGE-red cell membrane protein, AGE-aldose reductase, AGE-ferritin, AGE-red cell spectrin, AGE-alcohol dehydrogenase, AGE- haptoglobin, AGE-tubulin, AGE-thyroid hormone, AGE-fibrinogen, AGE- ₂- microglobulin, AGE-sorbitol dehydrogenase, AGE-arantitrypsin, AGE-carbonate dehydratase, AGE-RNAse, AGE-low density lipoprotein, AGE-hexokinase, AGE-apo C-l, and mixtures thereof.

3. The vaccine composition of any of the preceding claims, wherein the AGE antigen is an AGE-modified protein selected from the group consisting of AGE- RNAse, AGE-human hemoglobin, AGE-human serum albumin, AGE-low density lipoprotein, AGE-collagen IV, and mixtures thereof.

4. The vaccine composition of any of the preceding claims, wherein the AGE antigen is a condensation product advanced glycation end-product comprising a lysine component, an argine component and a reducing sugar component.

5. The vaccine composition of any of the preceding claims, wherein the AGE antigen is a compound of formula I:

wherein the lysyl component is indicated by the box labeled "K"; an arginyl component is indicated by the boxy labeled "R"; and a reducing sugar component is not boxed; and wherein R1 and R4 are independently H or an amide bond to an amino acid residue or a peptide chain; R2 and R3 are, independently, OH or an amide bond to an amino acid residue or a peptide chain; R5 is H, CH20H or CHOHCH20H; and wherein if more than one of R1 , R2, R3 or R4 is an amide bond, then the lysyl "K" component and the arginyl "R" component may be amino acid residues of the same or a different peptide chain.

6. The vaccine composition of any of the preceding claims, wherein the adjuvant comprises at least one member selected from the group consisting of aluminum phosphate, aluminum hydroxide, a water-in-oil emulsions, an oil-in-water emulsions, squalene, mannide oleate, a liposom, a vesicul, a paucilamellar vesicle, Mycobacterium bovis bacillus Calmette-Guerin, quill-saponin, unmethylated CpG dinucleotides, Freund's complete adjuvant and Freund's incomplete adjuvant.

7. The vaccine composition of any of the preceding claims, wherein the adjuvant comprises at least one member selected from the group consisting of aluminum phosphate, aluminum hydroxide, Freund's complete adjuvant and

Freund's incomplete adjuvant.

8. The vaccine composition of any of the preceding claims, wherein the preservative comprises at least one member selected from the group consisting of an antioxidant, an antibacterial agent and an antimicrobial agent.

9. The vaccine composition of any of the preceding claims, wherein the preservative comprises at least one member selected from the group consisting of benzethonium chloride, ethylenediamine-tetraacetic acid sodium, thimerosal, phenol, 2-phenoxyethanol, formaldehyde, formalin, amphotericin B, chlortetracycline, gentamicin, neomycin, polymyxin B, streptomycin, polyoxyethylene-9, 10-nonyl phenol, sodium deoxycholate and polyoxyethylated octyl phenol.

10. The vaccine composition of any of the preceding claims, wherein the composition is packed, sterilized, and in ready-to-inject form, and the composition is in unit dosage form containing the AGE antigen in an amount of 1 microgram to 100 milligrams.

11. The vaccine composition of any of the preceding claims, wherein the composition is in unit dosage form containing the AGE antigen in an amount of 10- 1000 micrograms.

12. The vaccine composition of any of the preceding claims, wherein the composition is packed, sterilized, and in ready-to-inject form,

the composition is in multidosage form containing 2-10 doses, and

each dosage contains the AGE antigen in an amount of 1 microgram to 100 milligrams.

13. The vaccine composition of any of the preceding claims, wherein each dosage contains the AGE antigen in an amount of 10-1000 micrograms.

1 . A method of treating atherosclerosis, comprising immunizing a human patient in need thereof against AGE-modified proteins.

15. The method of any of the preceding claims, wherein the immunizing comprises administering a vaccine comprising an AGE antigen.

16. The method of any of the preceding claims, wherein the vaccine comprises

(a) the AGE antigen,

(b) an adjuvant,

(c) a preservative, and

(d) optionally, an excipient.

17. A method of immunizing a human patient against AGE-modified proteins, comprising administering the vaccine composition of any of the preceding claims, in an amount effective to cause immunization against AGE-modified proteins.

18. A method of immunizing a human patient against AGE-modified proteins, comprising administering the vaccine composition of any of the preceding claims, in an amount effective to cause immunization against AGE-modified proteins.

19. The method of any of the preceding claims, wherein the vaccine composition is administered 2 to 10 times.

20. The method of any of the preceding claims, wherein the vaccine composition is administered 2 to 10 times over a time period of 1 week to 1 year.

21. The vaccine composition of any of the preceding claims, wherein the AGE antigen excludes AGE-modified albumin and AGE-modified collagen.