WO2010057242A2

WO2010057242A2 - Vaccine

Info

Publication number: WO2010057242A2
Application number: PCT/AT2009/000452
Authority: WO
Inventors: Sylvia Brunner; Pola Linzmayer-Hirt; Walter Schmidt; Bettina Wanko; Gabriele Winsauer; Christina Woess
Original assignee: Affiris Ag
Priority date: 2008-11-19
Filing date: 2009-11-19
Publication date: 2010-05-27
Also published as: AT507604A1; WO2010057242A3

Abstract

The present invention relates to a vaccine, comprising at least one peptide derived from the amino acid sequence TLGTNFGRCVDLFAPGEDII- GASSDCSTCFVSQSGTSQAAAHVAGIA (SEQ ID No. 1) and having a minimum length of 7 amino acids, wherein the peptide also comprises the amino acid sequence DIIGA (SEQ ID No. 2) and/or CFVSQSG (SEQ ID NO. 3) of SEQ ID No. 1.

Description

vaccine

The present invention relates to a vaccine for the treatment or prevention of health problems which may be caused by atherosclerosis, in particular stroke, cardiovascular diseases (which may lead to myocardial infarction), and peripheral vascular disease.

Diseases due to atherosclerosis, especially coronary heart disease, are the most common cause of death in the western world at -50%. Atherosclerosis is characterized, among other things, by the deposition of lipid particles in large arteries. In a multi-stage process that continues for many years to decades, atherosclerosis leads to narrowing of the affected vessels. Depending on the location of the vessels cardiovascular diseases (heart attack), stroke and peripheral vascular diseases may result. A major cause of atherosclerosis is lifestyle-related (low-exercise, high-fat diet, smoking) and / or genetic factors related to lipid metabolism, i. Too high total cholesterol or in particular to high levels of LDLc ("low density lipoprotein cholesterol") in combination with low HDLc values ("High Density Lipoprotein Cholesterol").

High LDLc values correlate directly with the occurrence of cardiovascular disease. The most common form of inherited high LDLc levels is autosomal dominant hypercholesterolemia (ADH). Related to this are three genes: the LDL receptor (LDLR), apolipoprotein B-100 (the ligand for the LDLR) and the protein convertingase PCSK9 ("proprotein convertase subtilisin kexin 9").

The family of secretory proprotein (PC) includes seven specific for basic amino acids subtilisin-like serine proteases ( "subtilisin-like serine proteina- ses ^w), namely PCl / 3, PC2, furin, PC4, PC5 / 6, PACE4 and PC7 , Also included are two other PC's that cleave at non-basic amino acids, namely SKI-1 / S1P and PCSK9 (also called NARC-I ("neural apoptosis-regulated convertase 1")).

With the exception of PC4, the convertases are expressed in the brain and peripheral organs. You have different radio For example, for the production of neuropeptides, growth factors, cytokines, receptors, in cell adhesion and cell migration as well as growth and differentiation of progenitor cells. Many convertases are known to play a role in diseases such as cancer or viral infections. Conditional shedding of PC5 / 6 in mice leads to malformations and bone defects in embryos, which is probably due to lack of processing of growth differentiation factor 11 ("growth differentiation factor 11").

Some members of the convertase family also affect the lipid metabolism:

- SKI-1 / S1P activates the synthesis of cholesterol and fatty acids as well as the LDL receptor (LDLR)

- Furin, PC5 and PACE4 inactivate endothelial lipase and lipoprotein lipase

PCSK9 inactivates the Low Density Lipoprotein Receptor (LDLR).

The gene for the human PCSK9 protein is located on chromosome Ip32.3. The gene is approximately 22 kilo base pairs long and encodes a 692 amino acid (AS) long glycoprotein. The expression of PCSK9 is regulated by sterol regulatory element binding proteins (SREBPs) and statins, which is also the case for other genes important in cholesterol metabolism. PCSK9 is mainly expressed in the liver but also in the small intestine and kidney and secreted. The 74 kDa pro-protein is autocatalytically processed in the endoplasmic reticulum, producing the approximately 60 kDa protein.

PCSK9 consists of

a 30 AS long signal peptide

the propeptide or inhibitor domain (AS 31-152)

- the "subtilisin-like catalytic domain" (AS 153-452) and

the C-terminal domain ("cysteine-rich unique C-terminal domain", CRD, AS 453-692).

PCSK9 plays a crucial role in cholesterol metabolism as it directly regulates the amount of LDLR present on liver cells. The LDLR is a glycoprotein located in the plasma membrane that removes cholesterol-rich LDLc particles from the plasma by endocytosis. The binding of PCSK9 leads to the uptake of LDLR into liver cells and subsequently its degradation into lysosomes as opposed to recycling to the cell surface that would occur without the binding of PCSK9.

In addition, PCSK9 seems to have an influence on the receptors for ApoE (ApoER2) and VLDL (Very Low Density Lipoprotein).

In vivo experiments in mice have shown that the amount of PCSK9 in human plasma is sufficient to reduce the amount of LDLR on the liver cells. In addition, several naturally occurring mutations in the PCSK9 gene have been identified in recent years that lead to increased ("gain-of-function mutations") or reduced ("loss-of-function mutations") activity of the protein.

Both in humans and in mice, gain of function mutations result in reduced levels of LDLR in the liver, causing high levels of plasma LDLc (hypercholesterolemia), predisposing to atherosclerotic coronary heart disease (coronary artery disease) , KHK) causes.

Over-expression of PCSK9 in mice leads to higher LDLc levels. Also, the administration of recombinantly produced PCSK9 protein dramatically reduces the number of LDL receptors in the mouse model.

"Loss of function mutations" cause higher levels of LDLR, resulting in lower LDLc levels, which proves to protect against coronary heart disease.

"Nonsense" mutations are common in some populations, eg, about 2% of African Americans have one of two mutations leading to a -30% reduction in LDLc levels, and a "missense" mutation is known in Caucasians which lowers the LDLc by ~ 15%. These lower LDLc levels lead to a significantly reduced incidence of CHD.

PCSK9 knock-out mice have more LDLR on their liver cells and reduced levels of plasma LDLc, and the inhibition of PCSK9 also leads to a significant reduction in total cholesterol and LDLc in mice fed on high-fat foods the amount of LDLR in the liver is doubled.

There are currently no known negative effects of a lack of PCSK9. _, Congenital homozygous "knock-out" of PCSK9 leads in humans to extremely low levels of LDLc with no noticeable side effects. The inhibition of PCSK9, for example, by antisense oligonucleotides, small molecule inhibitors or antibodies, is therefore an attractive target to reduce the effects of atherosclerosis, particularly cardiovascular disease, as a stand-alone therapy as well in combination with, for example, statins (which upregulate the PCSK9 protein) or with drugs that affect HDLc levels.

It is an object of the present invention to provide means that are suitable for reducing the content of PCKS9 and, as a result, of LDLc.

The present invention relates to a vaccine comprising at least one peptide with a minimum length of 7 amino acids derived from the amino acid sequence TLGTNFGRCVDLFAPGEDII- GASSDCSTCFVSQSGTSQAAAHVAGIA (SEQ ID NO: 1), wherein the peptide has the amino acid sequence DIIGA (SEQ ID NO: 2) and / or CFVSQSG (SEQ ID No. 3) of SEQ ID NO: 1.

It has been found that peptides of at least 7 amino acid residues derived from SEQ ID NO: 1 and comprising either SEQ ID NO: 2 or SEQ ID NO: 3 are capable of forming in a mammal and in a human of antibodies directed against PCSK9. The antibodies produced by administration are capable of binding to a portion of PCKS9 responsible for the binding of PCKS9 to the LDL receptor. This is to prevent the PCSK9-mediated degradation of LDLR and reduce the amount of LDLc, which in turn means that diseases that are the result of an elevated LDLc level (eg cardiovascular diseases), can be treated or treated as a preventive measure could be.

Peptides derived from SEQ ID NO: 1, but not SEQ ID NO: 2 or SEQ ID NO: 1. 3 surprisingly show the effects of the invention not or only inadequately.

The peptide according to the invention, which comprises both SEQ ID No. 2 and SEQ ID No. 3, likewise has the amino acid residues located between the two sequences and derived from SEQ ID No. 1.

The term "of the amino acid sequence TLGTNFGRCVDLFAPGEDII- GASSDCSTCFVSQSGTSQAAAHVAGIA (SEQ ID NO: 1) derived peptide "refers to peptide fragments of SEQ ID NO: 1 which are at least 7 amino acid residues in length, which peptides may also have a minimum length of 8, 9, 10, 11 or 12 amino acids Preferably, the peptides of the invention comprise at most 47, 45, 43, 40 or 35 amino acids Preferred peptides have a length of 8 to 20, more preferably 9 to 17, in particular 10 to 15, amino acids.

According to a particularly preferred embodiment of the present invention, the peptide is selected from the group consisting of IIGASSDCSTCFVSQSG (SEQ ID NO: 5), DLFAPGEDIIGASSDC (SEQ ID NO: 6), STCFVSQSGTSQAAAH (SEQ ID NO: 7), LFAPGEDIIGASSDC (SEQ ID NO: 5). 8), FAPGEDIIGASSDC (SEQ ID NO: 9), APGEDIIGASSDC (SEQ ID NO: 10), PGEDIIGAC (SEQ ID NO: 11), GEDIIGASSDC (SEQ ID NO: 12), EDIIGASSDC (SEQ ID NO: 13), DIIGASSDC (SEQ ID NO: 14), STCFVSQSGTSQAAA (SEQ ID NO: 15), STCFVSQSGTSQAA (SEQ ID NO: 16), STCFVSQSGTSQA (SEQ ID NO: 17), STCFVSQSGTSQ (SEQ ID NO: 18), STCFVSQSGTS (SEQ ID NO: 17). 19), STCFVSQSGT (SEQ ID NO: 20), STCFVSQSG (SEQ ID NO: 21), TCFVSQSGT (SEQ ID NO: 22), and CFVSQSG (SEQ ID NO: 23). In particular, the peptide STCFVSQSGTSQAAAH (SEQ ID NO: 7) and its C-terminal deletion derivatives having at least 9 amino acids (SEQ ID NO: 21) have been found to be particularly suitable.

The peptide according to the invention preferably has a cysteine residue at the C-terminus and / or at the N-terminus, a cysteine residue can e.g. also be added at the N and / or C terminus. Cysteine residues can be used, for example, for the cyclization of the peptides. Furthermore, further substances can be coupled to the SH groups of the cysteine residues, preferably to the N- or C-terminal cysteine residues. The cysteine residue may be a naturally occurring cysteine residue at this site or it may be attached at the N or C terminus of a sequence derived from the native sequence. Of course, it is also possible to couple via an internal cysteine.

According to a preferred embodiment of the present invention, the peptide according to the invention is coupled to a pharmaceutically acceptable carrier, preferably to KLH (keyhole limpet hemocyanin) (eg with NHS-PEO4 maleimides or other to those skilled in the ^art known suitable linkers)), tetanus toxoid, albumin binding protein, serum albumin, a dendrimer (MAP, Biol., Chem. 358: 581), peptide linkers (or flanking regions), and adjuvant substances described in Singh et al. , Nat. Biotech. 17 (1999): 1075-1081 (especially those in Table 1 of this document) and O 'Hagan et al. , Nature Reviews Drug Discovery 2 (9) (2003): 727-735 (in particular, the endogenous immunopotentiating compounds and delivery systems described therein) or mixtures thereof.

In addition, the vaccine composition can be formulated with an adjuvant, preferably a sparingly soluble aluminum composition, especially aluminum hydroxide. Of course, adjuvants such as MF59, aluminum phosphate, calcium phosphate, cytokines (eg IL-2, IL-12, GM-CSF), saponins (eg QS21), MDP derivatives, CpG oligos, LPS, MPL, polyphosphazenes, emulsions can also be used (eg, Freund's adjuvant, SAF), liposomes, virosomes, iscome, cochleates, PLG microparticles, poloxamer particles, virus-like particles, heat-labile enterotoxin (LT), cholera toxin (CT), mutant toxins (eg LTK63 and LTR72) , Microparticles and / or polymerized liposomes.

Other suitable adjuvants may be, for example, Purcell W et al. (Nature Reviews Drug Discovery 6 (2007): 404-414), especially Box 2.

The compound of the present invention is preferably linked to the carrier or adjuvant via a linker selected from the group consisting of NHS-poly (ethylene oxide) (PEO) (eg, NHS-PEO ₄ -maleimide). The conjugation chemistry (eg via heterobifunctional compounds, such as GMBS, and of course also others, as described, for example, in "Bioconjugate Techniques", Greg T. Hermanson) can in this context be selected from reactions known to the person skilled in the art Conjugation is preferably established via the N- or C-terminal cysteine residue (s).

According to a preferred embodiment of the present invention, the peptide is formulated for intradermal, subcutaneous or intramuscular administration to humans.

A vaccine comprising the peptide of the invention may be administered in any suitable manner, for example, id, iV, ip, im, intranasally, orally, subcutaneously, etc., and with any suitable delivery device (O'Hagan et al., Nature Reviews Drug Discovery 2 (9) (2003): 727-735). The peptide The present invention is particularly preferably formulated for intravenous, subcutaneous, intradermal or intramuscular administration (see, for example, "Handbook of Pharmaceutical Manufacturing Formulations", Sarfaraz Niazi, CRC Press Ine, 2004).

Typically, the vaccine will contain the peptide of the invention in an amount of 0.1 ng to 10 mg, preferably 10 ng to 1 mg, especially 100 ng to 100 μg, or alternatively, e.g. 100 fmol to 10 .mu.mol, preferably 10 pmol to 1 .mu.mol, in particular 100 pmol to 100 nmol. Typically, the vaccine may also contain adjuvant substances, e.g. Buffers, stabilizers, etc. included.

According to a preferred embodiment, the vaccine according to the invention is suitable for the treatment and / or prevention of cardiovascular diseases and other disorders caused by atherosclerosis, in particular cardiovascular diseases, strokes and peripheral vascular diseases (see Pschyrembel, Clinical Dictionary, 261).

Another aspect of the present patent application relates to a peptide as defined above.

The present invention relates not only to a vaccine of the type described above but also the peptides contained therein.

The peptides according to the invention are suitable for the treatment and / or prevention of diseases caused by atherosclerosis, in particular stroke, cardiovascular diseases and disorders of peripheral vessels.

Another aspect of the present invention relates to an isolated antibody directed against a peptide of the invention.

Instead of the peptides according to the invention, it is also possible to use antibodies which are capable of binding to these peptides. Such antibodies not only bind to the peptides but also to the PCSK9 protein present in the body.

The antibodies according to the present invention are preferably monoclonal antibodies. Such antibodies, which are homogeneous populations of antibodies to a particular antigen, can be obtained by any technique that allows the production of antibody molecules. These include, for example, the hybridoma technique of Kohler and Milstein (Nature 256 (1975): 495-497 and US Pat. No. 4,376,110), the human B-cell hybridoma technique (Kosbor et al., Immunology Today 4 (1983): 72; CoIe et al. , Proc. Natl. Acad. Be. USA 80 (1983): 2026-2030) and the EBV hybridoma technique (CoIe et al., Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., (1985): 77-96). Such antibodies may belong to any immunoglobulin class, including IgG, IgM, IgE, IgA, IgD, and any subclass thereof. The hybridoma producing the mAb of this invention can be cultured in vitro or in vivo. Of course, it is also possible to generate monoclonal antibodies by recombinant technologies in eukaryotic, yeast, insect and plant cells and in plants. These expression systems as well as the methods for isolating these antibodies from the cells are well known in the art. According to the invention, humanized monoclonal antibodies or functional parts thereof (Fab, single-chain antibodies, etc., which are able to recognize the peptides according to the invention) are particularly preferred.

A further aspect of the present invention relates to the use of a peptide or antibody according to the invention for the manufacture of a medicament for the treatment and / or prevention of disorders caused by atherosclerosis, in particular stroke, cardiovascular diseases and diseases of the peripheral vessels.

The present invention will be further illustrated in, but not limited to, the following figures and examples.

Fig. 1 shows the detection of PCK9-specific antibodies in mice after administration of a peptide having the amino acid sequence SEQ ID NO. 6 by ELISA.

Fig. 2 shows the detection of PCK9-specific antibodies in mice after administration of a peptide having the amino acid sequence SEQ ID NO: 27 by ELISA.

Figures 3 to 9 show the detection of PCK9-specific antibodies in mice after administration of a peptide having the amino acid sequences SEQ ID NOS: 8 to 14 by ELISA.

Fig. 10 shows the detection of PCK9-specific antibodies in mice after administration of a peptide having the amino acid sequence SEQ ID NO: 24 by ELISA.

Fig. 11 shows the detection of PCK9-specific antibodies in mice after administration of a peptide having the amino acid sequence SEQ ID NO: 7 by means of ELISA. Figures 12 to 20 show the detection of PCK9-specific antibodies in mice after administration of a peptide having the amino acid sequence SEQ ID Nos. 15 to 23 by means of EDISA.

Figure 21 shows the detection of PCK9-specific antibodies in mice after administration of a peptide having the amino acid sequence SEQ ID NO: 25 by ELISA.

Fig. 22 shows an ELISA in which purified mouse monoclonal antibodies generated after immunization with SEQ ID NO: 7 were added to the immunized peptide sequence as well as to truncations of this peptide (SEQ ID NOs: 21, 22, 23 and 25).

Samples: purified monoclonal antibody detection: α Mouse IgG

The table shows the raw ELISA data of Figure 22. The binding of monoclonal anti-PCSK9 antibodies to various peptides was tested in order to further determine the binding site of the antibodies.

No signal (background): OD less than 0.2

Weak signal: OD to 0.4

Medium signal: OD between 0.4 and 1.2

Strong signal: OD higher than 1.2

An antibody should preferably not only bind to the C-terminus, therefore all antibodies that recognize the tested peptides are in principle suitable.

Fig. 23 shows the titration of the monoclonal antibodies of Fig. 22 (immunization with p4918 = SEQ ID NO: 7). Samples: monoclonal antibody (start: 500 ng / ml) Detection: α mouse IgG In the table, the raw ELISA data are Figure 23 shown. The signal strengths of defined amounts of monoclonal anti-PCSK9 antibodies to recombinantly produced PCSK9 protein were compared in the ELISA, the signal strengths being reduced to a dilution of 15.6 ng / ml of antibody.

Weak signal: OD to 0.4

Medium signal: OD between 0.4 and 1.2 Strong signal: OD higher than 1.2

The highest possible signals are preferred (over OD 1.2 at an antibody dilution of 15.6 ng / ml).

Figures 24 and 25 show the detection of antibodies to the administered peptides (STCFVSQSGT-C, IIGASSDCSTCFVSQS, CIGIGDCSTCFVSQS, and IIGASSDCSTCFVSQS-C) in mice after peptide injection by peptide ELISA to measure antibody titers (Figure 24) ) and the detection of PCSK9-specific antibodies by PCSK9 protein ELISA in these sera (the sera were diluted 1: 100 and 1: 400, Fig. 25).

EXAMPLES:

Example 1:

1. Methods:

Production of vaccine:

The peptides were coupled to KLH via GMBS 4-maleimidobutyric acid N-hydroxysuccinimide ester. 30 μg of these conjugates (the amount refers to the peptide) were mixed with aluminum hydroxide (final Alum concentration 0.2%). The buffer used was PBS.

Immunization:

Balb / c or C57B1 / 6 mice were immunized subcutaneously (in the flank). Number of injections 2 to 4 times. The interval between immunizations was 1 week to 6 weeks. The injection volume per mouse was between 200 μl and 1 ml. Blood was drawn 2 weeks after immunization.

ELISA:

Serum or plasma of the immunized mice was tested by ELISA for antibodies against the immunized peptide and for antibodies to the PCSK9 protein.

Procedure:

The microtiter plates were coated with peptide (coupled to BSA) or protein

• The coated wells were incubated with serum or plasma at various dilutions

• the presence of peptide or PCKS9-specific antibodies in serum or plasma was detected by (preferably biotinylated) polyclonal anti-mouse IgG antibodies. The bound anti-mouse antibodies were then detected by incubation with streptavidin-HRP (horse radish peroxidase) (color reaction, e.g., ABTS as substrate).

2 results:

These results (see also Figures 1 to 23) show that the injection of peptides of the invention leads to the formation of PCKS9-specific antibodies. In contrast, the injection of the peptides with the sequences SEQ ID Nos. 24 and 25 showed no formation of PCKS9-specific antibodies.

Example 2:

Peptide ELISA (see Figure 22);

The said peptides were coupled to BSA and coated on ELISA plates. Subsequently, the monoclonal antibodies were added to the plates. The binding of antibodies to the peptides was standardized by means of biotinylated polyclonal anti-mouse IgG antibody (detection antibody) and subsequent parabolic reaction (streptavidin-HRP ("Horse Raddish peroxidase *); ABTS (2,2'-azino-bis (3 ethylbenzothiazoline-6-sulfonic acid) for the color reaction, measured at OD 405 nm).

After immunization of a mouse with SEQ ID NO: 7, various monoclonal antibodies had been generated. These antibodies were tested in the described ELISA for their formation on truncated peptide sequences derived from SEQ ID NO: 7 and on, respectively, to see if all antibodies are directed against the same amino acids of SEQ ID NO: 7, or if any of them for example, bind only at the N- or only at the C-terminus. This ELISA can be used to select for the following experiments antibodies that bind to certain amino acids or can not bind. It can thus be defined the binding site of the antibodies on the immunized peptide.

All tested antibodies bind strongly (OD higher than 1.2) to Seq ID Nos. 7, 21, 22 and 23. Differences only arise when bound to Seq ID No. 25. No binding (background; OD less than 0.2) is detectable in four of the antibodies (U1B1A4, U7F95H11, U10F11D4, U17G6C10), weak (OD to 0.4) binds one antibody (U4G5G8), medium binding (OD between 0.4 and 1.2 ) two antibodies (U9D8A2, U15E8C2).

It is apparent from this ELISA that binding of all the antibodies tested appears to be important in the presence of glutamine (AS No. 9 in SEQ ID NO: 7). The amino acids located C-terminal to this glutamine are apparently irrelevant to antibody binding. For all antibodies tested, the 7 amino acids of Seq. ID No. 23 to give a strong signal in the ELISA, i. the two amino acids at the N-terminus of Seq. ID No. 7 also does not seem to be essential in this analysis.

PCSK 9 Protein ELISA:

The monoclonal mouse anti-hu PCSK9 antibodies shown in Figure 22 were applied in different amounts (7.8 to 500 ng / ml respectively) to an ELISA plate coated with recombinantly produced PCSK9 protein. The detection of bound antibodies was carried out with biotinylated anti-mouse IgG and streptavidin-HRP. After addition of the substrate (ABTS), the OD was measured at 405 nm. The signal strength of the different antibodies was compared. The higher the signal, the stronger the antibody being tested binds to the PCSK9 protein. This ELISA can be used to select for subsequent experiments those antibodies which bind as strongly as possible to the target protein.

Analysis of the ELISA data:

As strongly binding antibodies in this ELISA those were designated, which exhibit an OD of more than 1.2 even with a dilution of 15.6 ng / ml, OD between 0.4 and 1.2 is designated as medium signal, among them as weak signal. Preferably, antibodies with the highest possible signal were used in further experiments. Example 3:

8 mice each were vaccinated with the peptides described in FIGS. 24 and 25 (coupled to KLH and mixed with alum) and the sera were analyzed after 1, 2, and 3 injections by means of peptide ELISA for the measurement of the antibody titer. The sera with comparably high anti-peptide titers were subsequently tested for PCSK9-specific antibodies by means of PCSK9 protein ELISA. investigated for the ability of the peptides to induce PCSK9-specific antibodies.

It was shown that the peptide STCFVSQSGT-C induces a stronger PCSK9-specific IRiran response than the peptides II-GASSDCSTCFVSQS, C-IIGASSDCSTCFVSQS, and IIGASSDCSTCFVSQS-C (see FIGS. 24 and 25). These results indicate that at least the C-terminal amino acid (G) of STCFVSQSGT-C is necessary to induce PCSK9-specific antibodies. This is also shown in the summary table under Example 1; the injection of SEQ ID NO: 20 (STCFVSQSGT) and SEQ ID NO: 21 (STCFVSQSG) lead to the formation of PCSK9-specific antibodies, while SEQ ID NO: 25 (STCFVSQS) does not induce PCSK9-specific antibodies.

Claims

Claims:

A vaccine comprising at least one peptide with a minimum length of 7 amino acids derived from the amino acid sequence TLGTNFGRCVDLFAPGEDIIGASSDCSTCFVSQSGTSQAAAHVAGIA (SEQ ID NO: 1), wherein the peptide has the amino acid sequence DIIGA (SEQ ID NO: 2) and / or CFVSQSG (SEQ ID NO: 3) SEQ ID NO: 1.

A vaccine according to claim 1, characterized in that the peptide is selected from the group consisting of II-GASSDCSTCFVSQSG (SEQ ID NO: 5), DLFAPGEDIIGASSDC (SEQ ID NO: 6), STCFVSQSGTSQAAAH (SEQ ID NO: 7), LFAPGEDIIGASSDC (SEQ ID NO: 8), FAPGEDIIGASSDC (SEQ ID NO: 9), APGEDIIGASSDC (SEQ ID NO: 10), PGEDIIGAC (SEQ ID NO: 11), GEDIIGASSDC (SEQ ID NO: 12), EDII-GASSDC (SEQ ID No. 13), DIIGASSDC (SEQ ID NO: 14), STCFVSQSGTSQAAA (SEQ ID NO: 15), STCFVSQSGTSQAA (SEQ ID NO: 16), STCFVSQSGTSQA (SEQ ID NO: 17), STCFVSQSGTSQ (SEQ ID NO: 18) , STCFVSQSGTS (SEQ ID NO: 19), STCFVSQSGT (SEQ ID NO: 20), STCFVSQSG (SEQ ID NO: 21), TCFVSQSGT (SEQ ID NO: 22), and CFVSQSG (SEQ ID NO: 23).

3. Vaccine according to claim 1 or 2, characterized in that the peptide has a cysteine residue at the C-terminus and / or at the N-terminus.

4. Vaccine according to one of claims 1 to 3, characterized in that the peptide is coupled to a pharmaceutically acceptable carrier, preferably to KLH (keyhole limpet hemocyanin).

5. A vaccine according to any one of claims 1 to 4, characterized in that the peptide is formulated for intradermal, subcutaneous or intramuscular administration.

6. A vaccine according to any one of claims 1 to 5, characterized in that the peptide is formulated with an adjuvant, preferably with aluminum hydroxide.

7. A vaccine according to any one of claims 1 to 6, characterized in that the peptide in an amount of 0, 1 ng to 10 mg, preferably 10 ng to 1 mg, in particular 100 ng to 100 micrograms, im Vaccine is included.

8. A vaccine according to any one of claims 1 to 7 for the treatment and / or prevention of disorders caused by atherosclerosis, in particular cardiovascular diseases, strokes or peripheral vascular diseases.

9. A peptide as defined in any one of claims 1 to 5.

10. A peptide according to claim 9 for the treatment and / or prevention of disorders caused by atherosclerosis, in particular cardiovascular diseases, strokes or peripheral vascular diseases.

11. An isolated antibody directed against a peptide according to claim 10.

12. Use of a peptide according to claim 9 or an antibody according to claim 11 for the manufacture of a medicament for the treatment and / or prevention of disorders caused by atherosclerosis, in particular cardiovascular diseases, strokes or peripheral vascular diseases.