TW201350509A - Vaccine - Google Patents

Abstract

The present invention relates to a vaccine comprising at least one peptide consisting at least of the amino acid sequence (X3)mKDX2QLGX1 (SEQ ID NO: 99), wherein X1 is an amino acid residue selected from the group consisting of alanine, asparagines, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, serine, threonine, tyrosine and valine, X2 is an amino acid residue selected from the group consisting of alanine, arginine, histidine, isoleucine, leucine, lysine, methionine, threonine, tyrosine and valine. X3 is (X4)nANISX5 (SEQ ID NO: 100) or an N-terminal truncated fragment thereof consisting of 1 to 4 amino residues, X4 is VVASQLR (SEQ ID NO: 101) or an N-terminal truncated fragment thereof consisting of 1 to 6 amino acid residues, X5 is an amino acid residue selected from the group consisting of alanine, asparagines, glutamine, glutamic acid, histidine, arginine, isoleucine, lysine, methionine, serine and threonine, m is 0 or 1, and n is 0 or 1, wherein said at least one peptide is coupled or fused to a carrier comprising at least one T-cell epitope.

Description

vaccine

The present invention relates to a medicament for use in the fields of medicine, immunity and molecular biology, which can prevent and/or treat chronic immune diseases caused by complement C5a.

The complement system is the central component of the endogenous immune system, protecting organisms from viruses, bacteria, and other foreign or abnormal cells. However, abnormal or excessive activation of the complement system may result in harm to the organism itself. Destructive capacity, unregulated activation of the complement system involves Alzheimer's disease, Parkinson's disease, Huntington's disease, age-related macular degeneration ( Age-related macula degeneration), rheumatoid arthritis, systemic lupus erythmatosus, antiphospholipid syndrome, asthma, vasculitis, atherosclerosis (atherosclerosis), multiple sclerosis, psoriasis, and chronic urticaria such as inflammatory dermatitis, Guillain-Barre syndrome, and hemolytic uremic syndrome (hemolytic uremic syndrome) and other chronic inflammatory diseases.

Unregulated activation of the complement system occurs in cancer, pregnancy, and antiphospholipid syndrome, such as pregnancy complication, sepsis, and acute lung injury. Acute pathological conditions such as acute respiratory distress syndrome (ARDS) and ischemia-reperfusion injury. Supplementary system A large number of activations occur more on artificial surfaces, causing hemodialysis-associated thrombosis.

The toxicity observed in the above state can be attributed to the overproduction of anaphylatoxin C5a to promote the immune response and to make the immune response permanent. The main function of C5a is chemotaxis and activated granulocytes. (granulocyte), mast cells, and macrophage, which release soluble immune factors. Therefore, in recent years, inhibition or regulation of the activity of the complement system has been considered to be a therapeutic strategy.

Among the complement proteins present in plasma, most of which are inactive precursors, which are cleaved and activated by a three-dimensional mechanism through the following three different mechanisms, wherein the mechanism comprises : typical pathways, lectin-inducing pathways and alternative pathways, the final result of which is the amplification of the reaction and the anaerobic toxins C3a, C5a and membrane attack complexes that kill cells. MAC) causes cells to create pores and cause cell hydrolysis.

The complement system consists of a C5 line 190kDa protein containing a dipeptide chain (α is 115kDa and β is 75kDa). Activation of either complement system pathway can generate C5 convertase, which in turn cleaves C5, making the new carboxy terminus of the C5a fragment The epitope (neoepitode) is exposed to produce C5b and a potent anaphylatoxin C5a.

Human C5a is a glycoprotein of 74 amino acids with a molecular weight of 12 to 14.5 kDa. The molecule consists of a tetra-alpha helix and stabilizes its structure with a three endogenous disulfide bond. Among them, aspartame, which is located at the 64th amino acid and has an N-linked carbohydrate moiety, is not essential for its biological activity, but is likely to regulate C5a activity in vivo. When the C5 convertase cleaves the C5a fragment, the arginine residue at the most carboxy-terminal end of the 74th amino acid of the C5a protein is rapidly removed by the serum and cell surface carboxypeptidase, and is less The active C5a desARG molecule, the C5a ARG and C5a desARG forms of the C5a protein, can bind to the seven-transmembrane domain receptor C5aR (CD88), and are not easily C5L2 (gpr77). It is recognized that C5L2 (gpr77) is commonly expressed in most cells, and is particularly expressed on the cell surface of immune cells such as macrophages, neutrophils, mast cells and T cells, and the ligand binding position of C5aR ( The ligand-binding site is complex and comprises at least two physically separable binding domains, one of which binds to a C5a disulfide-linked core (ie, amino acid 15 to 46), and the other C5a carboxy terminus (ie , amino acid 67~74) combined. The binding affinity of C5aR to its ligand C5a is excellent, and its dissociation constant (K _D ) is about 1 nM.

It is an object of the present invention to provide a means and method for treating a complement component C5a resulting in a disease or condition.

In order to achieve the foregoing object, the technical means and the effects achievable by the technical method of the present invention include: the present invention relates to a vaccine comprising at least one peptide, the peptide comprising 7-19 An amino acid residue, preferably comprising 7 to 14 amino acid residues, and the amino acid sequence of the peptide is (X ₃ ) _m KDX ₂ QLGX ₁ (SEQ ID NO: 99), wherein The X ₁ -monoamine acid residue is selected from the group consisting of alanine, aspartic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methyl sulfide a group consisting of aminic acid, serine, threonine, tyrosine, and valine, the X ₂ -amino acid residue selected from the group consisting of alanine, arginine, histidine, and iso-white the group consisting of alanine, leucine, lysine, methionine, threonine, tyrosine and valine, the amino acid sequence X ₃ is based (X _{4) n} ANISX ₅ ( SEQ ID NO: 100) or an amino terminal deletion fragment of the amino acid sequence comprising 1 to 4, preferably 1, 2, 3 or 4 amino acid residues, the X ₄ line being an amino acid sequence VVASQLR (shown in SEQ ID NO: 101) or contains 1 to 6, 5 or 6 for the good of the amino acid sequence of amino acid residues of the amino terminal deletion fragment, the amino acid residues X ₅ train a group selected from alanine, asparagine amine a group consisting of acid, glutamine, glutamine, histidine, arginine, isoleucine, lysine, methionine, serine, and threonine, wherein m is 0 or 1, and n is 0 or 1, wherein the at least one peptide is coupled or fused to a carrier, and the vector comprises at least one T cell epitope.

The present invention relates to an activating immune response against C5a of an organism itself, wherein the C5a of the organism itself is activated in a plurality of chronic immune diseases, and the immunological standard of the present invention forms an accessible state by cleavage of the C5 molecule. A new epitope at the carboxy terminus of C5a. Therefore, the C5b fragment that plays an important role in the defense process of the organism will not be affected by its generation and function.

In particular, the present invention relates to a vaccine which is a pro-hC5a carboxy-terminal epitope peptide (also known as VARIOTOPE) coupled or fused to a vector comprising at least one T cell epitope. The basis, wherein the peptide variant comprises a substitution of at least one amino acid residue in the procarboxy terminus of hC5a.

Interested peptide variants are not identical in antigenic epitopes, so the advantage of this peptide variant vaccine is to avoid the autotolerance of autoantigens. In addition, the use of peptide variants can be slowed down. Unintentional side effects caused by autoantigens.

The peptide variant of the hC5a carboxy-terminal epitope can be identified and selected by alanine scanning, which involves systematically replacing certain proteins with alanine residues or Individual amino acids in the peptide region to determine the functional role at certain positions, alanine is alternatively replaced by a residue, since alanine does not have a side chain of beta carbon atoms and does not alter the main chain structure Type, it will not seriously affect the electrostatic force or three-dimensional structure.

According to the above technique, it is possible to discriminate whether or not the amino acid residue of the hC5a carboxy terminal epitope is important for the humoral immune response caused by hC5a. In the next step, the replacement will not attenuate the humoral immune response induced by hC5a. Equivalent positions can be systematically replaced with amino acid residues having different characteristics to identify peptide variants that are capable of eliciting higher or at least equivalent anti-hC5a inhibitory activity compared to the original antigenic epitope. Next, the di- or tri-amino acid was simultaneously replaced at the carboxy-terminal epitope of hC5a to test its immunological and functional activity. A peptide variant having an antibody that induces a higher or at least equivalent anti-hC5a inhibitory activity is the subject of the invention as compared to the original carboxyl terminal epitope.

Surprisingly, only the peptide sequence as shown in SEQ ID NO: 99 is included, wherein at least positions 2, 3, 5, 6 and 7 are lysine, aspartate, and glutamine, respectively. Amine, leucine and glycine residues are sufficient to activate a good immune response (as shown in Figures 1A and 1B); more surprisingly, the amino acid in the last position of the peptide is At the time of substitution (such as X _{1 of} SEQ ID NO: 99, wherein the X ₁ arginine residue of the carboxy terminal region of the wild species C5a, refer to SEQ ID NOs: 1-4), that is, a body fluid sufficient to elicit anti-C5a protein The sexual immune response is significantly higher than the immune response elicited by the C5a fragment of wild species comprising SEQ ID NOs: 1-4 (see Figures 1A-1D). Importantly, as shown in Figures 2A to 2D, the more pronounced the humoral immune response, the more pronounced its inhibitory activity against C5a. When the penultimate position 5 of the wild-type C5a carboxy-terminal region is replaced with methylamine sulfate (such as SEQ ID NOs: 10 or 18) or when the wild-type C5a carboxy-terminal epitope is reciprocated to the eighth position of the histamine ( Similar results can be observed when the peptide of SEQ ID NOs: 7 or 17) is used for immunization (see Figures 1 and 2). From the above amino acids (for example, arginine, methionine and histidine in the first, fifth and eighth positions of the wild type C5a terminal epitope, please refer to SEQ ID NOs: 1-4 The single amino acid substitution can effectively elicit a C5a-specific antibody immune response, and the specific antibody immunoreaction is higher and more effective than the wild-type C5a fragment comprising SEQ ID NOs: 1-4 (please refer to the third As shown in Fig. 6, the peptide here has better C5a inhibitory ability than the wild type C5a fragment.

The WO 90/09162 patent unrelated to the present invention discloses several peptides homologous to the wild-type hC5a fragment, which are used as antagonists of C5a activity. Only, these Since the peptide is a soluble peptide and is not coupled to a carrier protein, it is not possible to induce antibody production against C5a activity, and is not suitable for use in the present invention. In the case of the 426th example of the WO 90/09162 patent, the peptide comprises a phenylalanine residue substituted at the first position, and in the present invention, the substitution system causes a significant decrease in the C5a inhibitory activity ( As shown in SEQ ID NO: 54 and Figure 4).

The vaccine of the present invention comprises at least one peptide comprising 7, preferably 8, preferably 9, preferably 10, preferably 11, preferably 12, preferably 13, preferably 14. Preferably, it is 15, preferably 16, preferably 17, preferably 18, preferably 19 amino acid residue.

According to the present invention, X ₃ is (X ₄ ) _n ANISX ₅ (as shown in SEQ ID NO: 100) or an amino terminal deletion fragment of the amino acid sequence, and therefore, the amino terminal deletion fragment may comprise ANISX ₅ ( As shown in SEQ ID NO: 102), NISX ₅ (as shown in SEQ ID NO: 103), ISX ₅ , SX ₅ or X ₅ ; that is, when m is 1, the vaccine system of the present invention may comprise an amine group. The acid sequence is ANISX ₅ KDX ₂ QLGX ₁ (as shown in SEQ ID NO: 104), NISX ₅ KDX ₂ QLGX ₁ (as shown in SEQ ID NO: 105), ISX ₅ KDX ₂ QLGX ₁ (eg SEQ ID NO: 106) Shown), at least one peptide of SX ₅ KDX ₂ QLGX ₁ (shown as SEQ ID NO: 107) or X ₅ KDX ₂ QLGX ₁ (shown as SEQ ID NO: 108).

According to the invention, X ₄ is a VVASQLR (as set forth in SEQ ID NO: 101) or an amino terminal deletion fragment of the amino acid sequence, and the VVASQLR fragment may comprise at least one amino acid selected from the group consisting of the following amino acid sequences Sequence: VASQLR (as shown in SEQ ID NO: 109), ASQLR (as shown in SEQ ID NO: 110), SQLR (as shown in SEQ ID NO: 111), QLR, LR, or R, m and n are 1, and the vaccine of the present invention may comprise a VAMSQLRANISX ₅ KDX ₂ QLGX ₁ (as shown in SEQ ID NO: 112), VASQLRANISX ₅ KDX ₂ QLGX ₁ (as SEQ ID NO: 113). Show), ASQLRANISX ₅ KDX ₂ QLGX ₁ (as shown in SEQ ID NO: 114), SQLRANISX ₅ KDX ₂ QLGX ₁ (as shown in SEQ ID NO: 115), QLRANISX ₅ KDX ₂ QLGX ₁ (eg SEQ ID NO: 116) Shown), LRANISX ₅ KDX ₂ QLGX ₁ (shown as SEQ ID NO: 117) or RANISX ₅ KDX ₂ QLGX ₁ (shown as SEQ ID NO: 118).

The vaccine of the present invention may comprise at least one peptide as set forth in SEQ ID NO: 99, in particular that the vaccine comprises at least one peptide having an amino acid sequence as set forth in SEQ ID NO: 99; However, the vaccine of the present invention may also comprise at least two, at least three, at least four, or even at least five peptides, and the at least two, at least three, at least four, or even at least five peptides have SEQ ID NO: 99 The amino acid sequence shown. The vaccine may of course also be combined with at least one peptide of the invention and other peptides or active ingredients, and the other peptides or active ingredients can be used to treat certain conditions of the invention.

The peptide/vector binding is important because when the peptide of the present invention not coupled to the carrier is injected into the body, an appropriate amount of the antibody cannot be induced, and further, the carrier can promote a prolonged antibody reaction; Therefore, the anti-hC5a active immune response of the present invention is superior to the single antibody antibody treatment for treating C5a-related diseases, thereby eliminating the need for repeated injections of large amounts of antibodies, frequent patient access to hospitals, and high production costs of antibodies such as C5a. The shortcomings of antibodies to therapy.

The at least one peptide of the present invention is capable of synthesizing a separate peptide by chemical synthesis, or a part of a peptide or a multi-peptide, which is generally in the technical field of the present invention. It is well known to the skilled person; the at least one peptide can also be a microorganism such as a bacterium, a yeast or a fungus, or a eukaryotic cell such as a mammalian cell or an insect cell, or an adenovirus or a pox virus. (poxvirus), herpesvirus, Semliki forest virus, baculovirus, bacteriophage, sindbis virus or sendai virus The recombinant viral vector produces the compound/peptide and is isolated or further purified. Bacteria suitable for the production of the compound/peptide include E. coli , B. subtilis or other bacteria which can express the peptide; suitable for expressing the compound/peptide The yeast type includes Saccharomyces cerevisiae , Schizosaccharomyces pombe , Candida spp. , Pichia pastoris or other yeasts which can express the peptides, the above method It is well known to those of ordinary skill in the art to which the present invention pertains, and methods for isolating and purifying recombinant peptides are also well known to those of ordinary skill in the art to which the present invention includes gel chromatography. (gel filtration), affinity chromatography, ion exchange chromatography, and the like.

To aid in the isolation of the compound/peptide, a fusion multi-peptide can be prepared, wherein the compound/peptide is fused (covalently linked) to a heterologous multi-peptide, which is heterologous It can be isolated by affinity chromatography. Typical heterologous peptides are histidine tags (such as His6, 6 histidine residues), GST tag (Glutathione-S-transferase), etc. The system not only helps the purification of the compound/peptide, but also prevents the compound/peptide from being degraded during the purification process. When the fusion multi-peptide is pre-removed after purification, the fusion multi-peptide may further comprise a cleavage site comprising an amino acid sequence at the compound/peptide and the junction thereof. And the amino acid sequence may be cleaved by an enzyme (eg, a protease) specific for the amino acid sequence.

X ₁ may be a non-polar, aliphatic amino acid residue such as alanine, glycine, valine, leucine, methionine or isoleucine, or may be as serine a polar, uncharged amino acid residue such as sulphate, aspartame or glutamine, or may be a positively charged amino acid residue such as lysine or histidine, or may be Like polar, aromatic amino acid residues such as tyrosine. X ₂ may be an amino acid residue selected from the group consisting of alanine, methionine, valine, leucine, isoleucine, lysine, arginine, histidine, threonine and A group consisting of tyrosine. X ₅ may be a non-polar, aliphatic amino acid residue such as alanine, methionine or isoleucine, and may be, for example, a serine, a sulphate, an aspartate or a glutamine. A non-polar, aliphatic amino acid residue such as an amine, or may be a charged amino acid residue such as lysine, arginine, histidine, glutamic acid or the like.

In a preferred embodiment of the invention, the X ₁ -monoamine acid residue is selected from the group consisting of threonine, glutamine, tyrosine, methionine, alanine, glycine, and valine The group that makes up.

In a preferred embodiment of the invention, when m is 1, X _{1 is} preferably an amino acid residue selected from the group consisting of alanine, aspartame, glutamine, histidine, lysine, and A. a group consisting of thiaminic acid, serine acid, and threonine, and an X ₅ -monoamine acid residue selected from the group consisting of alanine, histidine, methionine, and threonine And/or an X ₂ -monoamine acid residue selected from the group consisting of methionine, alanine, lysine, and valine.

In a preferred embodiment of the invention, when m is 1, X _{5 is} preferably an amino acid residue selected from the group consisting of alanine, methionine and threonine.

In a preferred embodiment of the invention, the X ₂ -monoamine acid residue is selected from the group consisting of methionine, alanine, lysine, and valine.

In a preferred embodiment of the invention, the at least one peptide is preferably selected from the group consisting of ISHKDMQLGA (as shown in SEQ ID NO: 14), ANISHKDMQLGA (shown as SEQ ID NO: 21), and KDMQLGA (such as SEQ ID NO: 22)), VVASQLRANISHKDMQLGA (as shown in SEQ ID NO: 23), ANISHKDMQLGT (as shown in SEQ ID NO: 24), ANISHKDMQLGQ (as shown in SEQ ID NO: 25), and ANISHKDMQLGY (as in SEQ ID NO: 26) Shown, ANISHKDMQLGM (as shown in SEQ ID NO: 27), ANISHKDMQLGG (shown as SEQ ID NO: 28), ANISHKDMQLGV (shown as SEQ ID NO: 29), ANISHKDMQLGK (shown as SEQ ID NO: 30) , AISHIKDMQLGS (shown as SEQ ID NO: 31), ANISHK DMQLGH (shown as SEQ ID NO: 32), ANISHK DMQLGN (shown as SEQ ID NO: 33), ANISHK DMQLGL (shown as SEQ ID NO: 34), ANISTKDMQLGA (shown as SEQ ID NO 70), ANISTKDMQLGQ (shown as SEQ ID NO: 71), ANISTKDMQLGS (shown as SEQ ID NO: 72), ANISTKDMQLGM (shown as SEQ ID NO: 73), ANISMKDMQLGN (such as SEQ ID NO: 74), ANISTKDKQLGM (shown as SEQ ID NO: 75), ANISTKDMQLGH (shown as SEQ ID NO: 76), ANISAKDMQLGA (shown as SEQ ID NO: 77), ANISMKDMQLGA (eg SEQ ID NO: SEQ ID NO: :78), ANISTKDKQLGA (as shown in SEQ ID NO: 79), ANISTKDAQLGA (as shown in SEQ ID NO: 80), ANISMKDMQLGS (as shown in SEQ ID NO: 81), ANISTKDVQLGA (such as SEQ ID NO: 82) Shown), ANISTKDMQLGN (shown as SEQ ID NO: 83), ANISTKDMQLGK (shown as SEQ ID NO: 84), ANISMKDMQLGM (shown as SEQ ID NO: 85), ANISTKDMQLGT (shown as SEQ ID NO: 86) , a group consisting of ANISHKDKQLGK (shown as SEQ ID NO: 87), ANISMKDMQLGH (shown as SEQ ID NO: 88), and ANISAKDAQLGA (shown as SEQ ID NO: 89).

In a preferred embodiment of the invention, the at least one peptide comprising an amino acid sequence as set forth in SEQ ID NO: 99 is preferably contained directly at or at an amino terminus and/or carboxyl terminus. The spacer sequence binds at least half of the cysteine residues.

The cysteine residue can serve as a reaction group such that the peptide can be combined with other molecules or a carrier; for example, the reaction group can be used to bind the peptide to a carrier protein. The cysteine residue may be directly bound to the peptide or combined with the peptide via a spacer sequence, preferably comprising at least one, preferably at least two, more preferably at least three, particularly It is a small, non-polar amino acid residue such as glycine at least four, the maximum of which is ten, preferably a minimum of five.

In a preferred embodiment of the invention, the vector is selected from the group consisting of keyhole limpet haemocyanin (KLH), CRM197, tetanus toxoid (tetanus) Toxoid, TT), diphtheria toxin (DT), protein D or other proteins or peptides containing T cell epitopes.

In the present invention, the peptide is coupled or fused to a pharmaceutically acceptable carrier, which is keyhole limpet hemocyanin (KLH), tetanus toxoid, aluminum binding protein (albumin-binding). Protein), bovine serum albumin, (dendrimer), peptide? (peptide linker, or flanking region), or as Singh et al. (see Singh et al. , Nat. Biotech. 17, (1999): 1075~1081, Table 1) and O'Hagan et al (please refer to O'Hagan and Valiante, Nature Reviews, Drug Discovery 2 (9); (2003): 727-735, 〝endogenous immuno-potentiating compounds and delivery systems, adjuvants, or mixtures thereof. The chemical process in which the peptide is coupled or fused to the carrier (for example, by GMBS and other "Bioconjugate Techniques", a heterobifunctional compound published by Greg T. Hermanson) can be selected as the process of the chemical reaction. The field of the invention is known to those of ordinary skill in the art.

At least one peptide of the present invention may also be fused to a protein carrier by a method known to those of ordinary skill in the art, such protein comprising a peptide and an unrelated immune genetic protein, preferably, The immune genetic protein system can induce a recall response. For example, the protein system can contain tetanus, tuberculosis, hepatitis protein and protein D, and is a Gram-negative bacteria type B. influenzae. Surface protein of Haemophilus influenza B) (refer to WO 91/18926); preferably, a protein containing about one-third of the protein D (for example, the amino-terminal 100-110 amino acid) can be used. A derivative, and the derivative can be lipidate. The vector which can be used as the fusion protein may be, for example, a LYTA protein, or a fragment of LYTA (preferably the carboxy terminus of LYTA), LYTA is derived from Streptococcus pneumoniae, and the synthesis is called guanamine LYTA ( Amidase LYTA) N-acetyl-L-alanine amidase (N-acetyl-L-alanine amidase, (decoded from LytA gene, please refer to Gene 43; (1986): 265-292), LYTA is an autolysin that specifically degrades certain bonds of peptidoglycan bone price. In a preferred embodiment of the invention, a fusion protein comprises a repeat portion of LYTA, the repeat portion being at the carboxy terminus and starting at residue 178, preferably selected from residues 188-305.

In a preferred embodiment of the invention, the peptide is complexed with an adjuvant, preferably with aluminum.

The vaccine of the present invention may be compounded with an adjuvant which is preferably a low solubility aluminum composition, particularly aluminum hydroxide. Of course, the adjuvant may also be MF59 or aluminum phosphate. (aluminum phosphate), calcium phosphate, cytokines (cytokine (eg, IL-2, IL-12, GM-CSF), saponin (eg, QS21), MDP derivatives, oligo CpG (CpG) Oligo), LPS, MPL, polyphosphazene, emulsion (eg, Freund's adjuvant, SAF), liposome, virosome, immunostimulating complex (iscom), Spiral liposome (cochleate), PLG microparticles, poloxamer particles, virus-like particles, heat-labile enterotoxin (LT), cholera toxin (cholera toxin) , CT), mutant toxins (eg, LTK63 and LTR72), microparticles and/or polymeric lipid vesicles.

Suitable adjuvants are commercially available adjuvants such as, for example, AS01B, AS02A, AS15, AS-2 and its derivatives (GlaxoSmithKline, Philadelphia, PA), CWS, TDM, Leif, aluminum hydroxide gel Aluminium salt such as (alum) or aluminum phosphate, calcium salt, iron or zinc, insoluble suspension of acetaminophen tyrosine, acylated sugar, polysaccharide derivative with anion or cation, polyphosphazene, Biodegradable microsphere, monophosphoryl lipid A and quil A, in addition, cytokines such as GM-CSF or interleukin-2, interleukin-7 or interleukin-12 It can also be used as the adjuvant.

In the vaccine, the adjuvant composition preferably induces an excellent Th1 type immune response, and a high amount of Th1 type cytokines (eg, IFN-y, TNFct, IL-2, and IL-12) can assist in stimulating A cell mediated immune response is administered to the administered antigen; relatively, a high amount of Th2 type cytokines (eg, IL-4, IL-5, IL-6, and IL-10) will assist Induction of a humoral immune response.

After the vaccine is administered, the patient will develop an immune response comprising a Th1 type and a Th2 type. In the preferred embodiment, the system exhibits a more pronounced Th1 type immune response, which increases the Th1 type cytokine content significantly higher than the Th2 type cell. Hormone content, these cytokine levels can be analyzed by standard analytical methods. Recalling the plurality of the cytokine family, see Janeway et al., Immunobiology, 5 th Edition, 2001 a.

A preferred adjuvant may elicit a significant Th1 type reaction, and the adjuvant may comprise, for example, a monophosphorus lipid A composition, preferably a 3-O-deacylated monophosphoryl lipid A (3-O-deacylated monophosphoryl lipid) A, 3D-MPL) can also be used in combination with an aluminum salt (for example, as Ribi et al., Immunology and Immunopharmacology of Bacterial Endotoxins, Plenum Publ. Corp., NY, (1986): 407-419, or GB 2122204B, GB 2220211 and US 4,912,094, etc.), the preferred use form of 3D-MPL is an emulsion, wherein the particle size is less than 0.2 mm (diameter), and the preparation method thereof is disclosed in WO 94/21292 patent; Liquid formulations of monophosphoryl lipid A and a surfactant have been disclosed in the WO 98/43670 patent. Preferred adjuvants in the examples include AS01B (MPL and QS21 in a liposome formulation), 3D-MPL and QS21 in a liposome formulation, AS02A (MPL and QS21 and an oil-in-water emulsion), 3D-MPL and QS21 and an oil-in-water emulsion, and AS 15 (available from GlaxoSmithKline) (MPL adjuvant is available from GlaxoSmithKline, Seattle, WA) (see U.S. Patent Nos. 4,436,727, 4,877,611, 4,866,034, and 4,912,094).

CpG-containing oligonucleotides (whose CpG dinucleotides are not methylated) can also induce significant Th1 immune responses, CpG-based DNA cytosine-guanine dinucleotide Abbreviation for the structure (cytosine-guanosine dinucleotide motif), which is well known and mentioned in WO 96/02555, WO 99/33488, US 6,008,200 and US 5,856,462, etc., to promote the immunization of DNA sequences. As described in Sato et al., Science 273; (1996): 352, when CpG is made into a vaccine, it is usually dissolved in a buffer solution together with a free antigen (see WO 96/02555, McCluskie and Davis, supra) or covalently bound to an antigen (as shown in WO 98/16247), or with, for example, aluminum hydroxide (Hepatitis surface antigen, please refer to Davis et al., supra; Brazolot-Millan et al., PNAS USA, 95 (26), (1998): 15553-8), etc., after a carrier is compounded and administered. Furthermore, CpG is preferably administered in a systemic or mucosal route, as is known to those of ordinary skill in the art to which the present invention pertains (e.g., WO 96/02555, EP 0 468 520, Davis et al., J. Immunol, 160(2), (1998): 870~876 and McCluskie and Davis, J. Immunol., 161(9), (1998): 4463-6).

Another preferred adjuvant is a saponin or saponin analog or derivative, preferably QS21 (Aquila Biopharmaceuticals Inc.), which may be used alone or in combination with other adjuvants. For example, an enhancement system comprises a monophosphoryl lipid A and a saponin derivative, such as the QS21 and 3D-MPL compositions described in the WO 94/00153 patent, or as described in the WO 96/33739 patent. One of the less reactive compositions of QS21 bound by cholesterol. The remaining preferred formulations comprise an oil-in-water emulsion and tocopherol, such as one of the preferred adjuvant formulations of WO 95/17210 comprising QS21, 3D-MPL and tocopherol in an oil-in-water emulsion. Further, the saponin adjuvant used in the present invention comprises QS7 (as described in WO 96/33739 and WO 96/11711 patents) and QS17 (as described in US Pat. No. 5,057,540 and EP 0 362 279 B1).

Another preferred adjuvant comprises Montanide ISA 720 (Seppic, France), SAF (Chiron, California, United States), immunostimulating complex (CSL), MF-59 (Chiron), SBAS series adjuvants (eg SBAS-2) , AS2', AS2, SBAS-4 or SBAS6, It is commercially available from GlaxoSmithKline), Detox (Corixa), RC-529 (Corixa, Hamilton, MT) and other amino-alkyl glucosaminide 4-phosphates (AGP). Further examples include synthesis. MPL and an adjuvant based on Shiga toxin B subunit (refer to WO 2005/112991 patent).

The vaccine of the present invention can be administered by subcutaneous injection, intramuscular injection, intradermal injection, intravenous injection or the like (please refer to "Handbook of Pharmaceutical Manufacturing Formulations", Sarfaraz Niazi, CRC Press Inc, 2004), and The medicament may comprise a carrier, an adjuvant and/or an excipient depending on the route of administration.

The vaccine of the present invention comprises 0.1 ng to 10 mg of the compound, preferably 10 ng to 1 mg, particularly 100 ng to 100 μg, or alternatively, 100 fmol to 10 μmol, preferably 10 pmol to 1 μmol, particularly 100 pmol to 100 nmol. The compound or peptide of the present invention is administered to a mammal at a dose of 100 ng to 1 mg each time, preferably 1 μg to 500 μg, more preferably 10 μg to 100 μg, particularly 20 to 40, or 30 μg. Typically, the vaccine may contain auxiliary substances such as buffer solutions, stabilizers and the like. The vaccine of the present invention can be administered 3 to 6 times every 2 to 2 months, and the vaccine is administered at intervals of about every 6 months due to the presence of anti-C5a antibodies.

AMD is a medical condition that usually affects the elderly. Because of retinopathy, the elderly have visual loss (ie, spots) in the center of the field of vision. AMD contains "dry form" and "wet type ( Wet form)", of which dry type accounts for about 90% of AMD patients. The early diagnosis of wet and dry AMD is an amorphous lipoproote inaceous deposit that accumulates around the epidermal cells of the retinal pigment. This pathological component is called drusen, a recent study. It is pointed out that the formation of a dry AMD index, the regional inflammatory response and the activation of the complement system, is consistent with other studies showing that the complement system component C5 is clustered in the concealed junction.

In addition, in addition to the release of VEGF, C5a also plays an important role in choroidal new vascularization induced by wet AMD. Most importantly, antagonizing C5a neutral antibodies can stop the disease in animal models. Development.

In summary, the complement system regulates disease in dry and wet AMD as a strong support, so C5a seems to be the best target for the treatment of dry and wet AMD.

The inflammatory response regulated by the complement system, mainly caused by C5a, is thought to play an important role in the acceleration and progression of Alzheimer's disease. In the brain of Alzheimer's disease, fibrillar Aβ plaque It leads to prolonged activation of the complement system, and many of the evidence for this disease comes from the glial cells (glia) recruited and activated by C5a to promote inflammatory responses. Similar phenomena apply to Parkinson's disease and Huntington's dance. disease. In addition, preliminary results indicate that the activated fragment (C5a) of the complement component C5 plays a specific role in motor neuron disease, which causes progressive death of motor neurons, which ultimately leads to degenerative diseases of sputum and death. The role of sexual pathogenicity.

The blockade of C5aR can significantly slow down the respiratory tract inflammation and airway hyper-responsiveness of allergic asthma. However, the role of complement component C5 in asthma is still controversial. In allergic asthma, C5 was thought to promote or prevent respiratory hyperreactivity in asthma, suggesting that C5a plays a dual role in allergic asthma; a hypothesis is that C5aR signaling in allergen sensitization prevents lung allergy ( Pulmonary allergy) occurs, but increases in the inflamed pulmonary environment of the effector phase, ie, blocking C5aR may have therapeutic advantages in established asthma.

C5a also plays a role in atherosclerosis. C3a and C5a are expressed in human coronary plaques. In addition, C5a is more recently predicted to predict cardiovascular events in patients with advanced atherosclerosis and C5a in serum. Rise After the balloon angioplasty with the superficial femoral artery, there is a correlation between the development of reocalosis.

Vasculitis is the inflammatory process of blood vessels. Its histopathological features are the inflammatory reaction of the blood vessel wall and fibrinoid necrosis. The clinical spectrum of this vasculitis can be puupura to severe. Proliferative glomerulonephritis, and the complement system is considered to be involved in these processes. For example, C5a plays an important role in anti-neutrophil cytoplasmic autoantibody (ANCA)-associated vasculitis, an anti-neutrophil cytoplasmic autoantibody-associated vasculitis is a relatively Uncommon, but potentially life-threatening, systemic autoimmune disease; complement system involvement in ANCA-induced necrotizing crescentic glomerulonephritis, C5a and the hobby The granulocyte C5aR may constitute an activation loop to activate ANCA-regulated neutrophil activation, which may provide a new therapeutic target for the treatment of necrotic crescentic glomerulonephritis caused by ANCA.

Activation of the complement system involves pathological changes in the inflammatory response of autoimmune dermatitis such as bullous pemphigoid (BP), psoriasis vulgaris, and chronic urticaria. In pemphigus, activation of the complement system caused by epithelial pemphigus antibodies may be responsible for the development of typical inflammatory response changes (also known as eosinophilic spongiosis); in dryness, high levels can be observed. C5a, showing that activation of the complement system is also involved in the disease, and the stem cell-T cell regulates the disease. However, neutrophils and mast cells may also be involved in the pathogenesis of the disease, and both T cells and neutrophils will be C5a is attracted, so C5a may be an important therapeutic target for the treatment of dryness.

Activation of the complement system also contributes to autoimmune diseases such as rheumatoid arthritis. The anaphylatoxin C5a is the main product of the activation of the complement system causing tissue damage in rheumatoid arthritis, although the breakdown of the membrane attack complex and the C3b fragment Opsonization It is also very important.

The role of the complement system in the pathogenesis of lupus erythematosus (SLE) remains controversial. On the one hand, the complement system component regulates the tissue damage initiated by autoantibodies. On the other hand, the complement system has some genetic defects. Protection features, while some complement systems are associated with increased risk of SLE, such as patients with SLE often have hypocomplementemia. In addition, the C5a/C5aP message transmission pathway plays an important role in the pathogenesis of the central nervous system lupus by regulating the integrity of the blood-brain barrier, and the C5a/C5aR resistance. Broken is considered a promising SLE treatment policy.

Tissue reperfusion (R), but not ischemia (Ischemia, I), activates the complement system and causes inflammatory damage, although activation of the complement system is involved in ischemia-reperfusion injury (I/R injury) Clearly, many studies have pointed out that the complement system is linked to the pathogenesis of ischemia-reperfusion injury, and speculates that inhibition of the complement system is a viable treatment strategy. For example, in a murine myocardial ischemia-reperfusion injury mode, a systemic C5 inhibition response 30 minutes prior to reperfusion can effectively prevent myocardial ischemia-reperfusion injury in mice.

Activation of the complement system has been demonstrated in a variety of acute lung injuries. Among the acute lung injury caused by infusion of acid, the C5a concentration of broncho-alveolar lavage fluids (BALF) increases, and the concentration of C5a also increases in human transplanted lungs. C5a attracts neutrophils into the lungs and directly activates neutrophils, macrophages and endothelial cells. The protective effect of anti-C5a is related to the dramatic decrease of TNF-α in BALF, and it is the same as that of ICAM-15, which is a significant decrease in pulmonary vascular cell adhesion molecule in BALF. It is speculated that the inflammatory response is regulated based on the inflammatory response network. C5a is necessary in the expression of regulatory factors and in the expression of adhesion factors.

Acute lung injury and acute respiratory distress syndrome (ARDS) are characterized by lung There are fibrin-rich inflammatory secretions in the intra-alveolar spaces, and alveolar cells that move neutrophils into the lungs extensively. In the neutrophils taken from healthy volunteers, drug-blocking of TNF-α and C5a signaling can significantly reduce the procoagulant activity of BALF induced by other healthy cells, accompanied by tissue factor (tissue factor, TF) loss of performance. These results indicate that the signaling of C5a and TNF-[alpha] contributes to the expression of neutrophil tissue factor accumulated in the alveoli of the lungs affected by ARDS.

In the course of sepsis, the inflammatory response system containing both cell and body fluid defense mechanisms is highly activated. It is known that in human sepsis, the activation of complement, especially the increase in C5a expression, is associated with a significantly reduced survival rate associated with multiple organ failure compared to less severe sepsis patients and survivors. Even blocking C5a or C5aR significantly increased the survival rate of sepsis-bearing rodents. Therefore, C5a seems to play a key role in the development of sepsis, and interference with C5a/C5aR may be a promising clinical direction to provide a prophylactic treatment to patients at high risk of developing sepsis.

In cardiopulmonary bypass and hemodialysis, when human blood comes into contact with the artificial surface of a heart-lung machine or kidney dialyzer, C5a is activated by an alternative complement pathway. C5a causes increased microvascular permeability and edema, bronchoconstriction, pulmonary vasoconstriction, activation of white blood cells and platelets, and infiltration into tissues (especially the lungs). Administration of monoclonal antibodies against C5a reduces coronary endothelial dysfunction caused by cardioplegia or cardioplegia.

Tumor-driven complement activation can provide benefits for tumor growth. The production of complement C5a in the tumor microenvironment can promote tumor growth by inhibiting the action of anti-CD8+ T cells. Tumor growth experiments using mice have shown that the lack or blockade of C5aR is associated with tumor growth retardation. Therefore, complement inhibition is considered to be effective and promising in cancer therapy. The direction.

Significant increases in complement activation are associated with different pathological pregnancy outcomes, such as preeclampsia, recurrent spontaneous abortion, intrauterine growth retardation, and antiphospholipid syndrome (APS). The concentration of C5a in the plasma of women with pregnancy toxemia is increased compared to normal women. In APS, activation of antiphospholipid antibodies and complement (via C3a, C5a, and MAC) may be combined with initiation of a local inflammatory response that ultimately leads to placental thrombosis, hypoxia, and neutrophil infiltration. In anti-phospholipid-induced fetal damage, tissue factor (TF) is associated between C5a and neutrophil activation.

In summary, the peptide reacts with an anti-C5a immune response, resulting in an effective treatment for C5a-regulated (chronic inflammation) diseases, including neurodegenerative diseases such as Alzheimer's disease (see, for example, Fonseca, MI et al. 2009), J Immunol "Treatment with a C5aR Antagonist Decreases Pathology and Enhances Behavioral Performance in Murine Models of Alzheimer's Disease." and Klos, A. et al. (2009), Mol Immunol "The role of the anaphylatoxins in health and disease. "), Parkinson's disease (see, eg, McGeer, PL et al. (2004), Parkinsonism Relat Disord "Inflammation and neurodegeneration in Parkinson's disease."), Huntington's disease (see, eg, Singhrao, SK et al. (1999) ), Exp Neurol "Increased complement biosynthesis by microglia and enhance activation on neurons in Huntington's disease."), and age-related macular degeneration (see, eg, Nozaki, M. et al. (2006), Proc Natl Acad Sci "Drusen complement components C3a and C5a promote choroidal neovascularization."), rheumatoid arthritis (see, for example, Okroj, M. et al. (2007), Ann Med "Rheumatoid Arthritis and the complement system."), lupus erythematosus (see, for example, Chen, M. et al. (2009), J Autoimmun "The complement system in systemic autoimmune disease."; Jacob, A. et al. (2010), J Neuroimmunol "Inhibition of C5a receptor alleviates experimental CNS lupus." and Jacob, A, et al. (2010), FASEB J "C5a alters blood-brain barrier integrity in experimental lupus."), asthma (see eg Kohl, J. et al. (2006), J Clin Invest "A regulatory role for the C5a anaphylatoxin in type 2 immunity in asthma."), vasculitis, antiphospholipid antibody syndrome (APS), atherosclerosis, inflammatory Dermatitis such as psoriasis and chronic urticaria, Guillain-Barre syndrome, hemolytic uremic syndrome, and multiple sclerosis. Loss of controlled hC5a release may cause pathological conditions such as ischemia, reperfusion injury, sepsis, acute lung injury, complex conditions of hemodialysis, tumors, complex conditions of pregnancy such as pregnancy toxemia and APS, by activating the immune system Neutralizing C5a may provide an effective therapy for these pathological complexities.

Figure 1 is an alanine scan of carboxy-terminal fragments of different lengths of hC5a (shown in SEQ ID NOs: 1-4) (shown in SEQ ID NOs: 5-23) for the purpose of defining replacement It does not lose immunity and triggers the amino acid position of the antibody against C5a. Wherein, Figure 1A is the position of the original epitope of hC5a (epitope) 65-74 (as shown in SEQ ID NO: 1) and the immunity of the peptide variant (described as titer); Figure 1B hC5a original antigen epitope amino acid position 63-74 (as shown in SEQ ID NO: 2) and VARIOTOPE immunity; 1C map hC5a original antigen epitope position 68-74 (such as SEQ ID NO: 3) and the immunity of VARIOTOPE; Figure 1D is the position of the original epitope of hC5a, 55-74 (as shown in SEQ ID NO: 4) and the immunity of VARIOTOPE.

Figure 2 is an inoculation peptide variant SEQ ID NOs: 5 to 23 mice immune serum inhibition activity The sequence of the original, epitope (as shown in SEQ ID NOs: 1-4) is described as 100%. Among them, the 2A map is the position of the original epitope of hC5a epitopes 65-74 (as shown in SEQ ID NO: 1) and the inhibitory effect of the immune serum induced by the peptide variant; the 2B map is the original epitope of hC5a 63 Position ~74 (as shown in SEQ ID NO: 2) and the immune serum inhibitory effect induced by the peptide variant; Figure 2C shows the position of the original epitope of hC5a at positions 68-74 (as shown in SEQ ID NO: 3) And the immune serum inhibitory effect by the peptide variant; the 2D map is the position of the original epitope of hC5a 55-74 (as shown in SEQ ID NO: 4) and the immune serum inhibitory effect induced by the peptide variant.

Figure 3 is an evaluation of the inhibitory activity of the antibody elicited by a peptide variant of 12 amino acid lengths at the carboxy terminus of hC5a (as shown in SEQ ID NO: 2), and the R of the amino acid position 74 of hC5a is replaced. Amino acid residues of different characteristics (as shown in SEQ ID NOs: 21, 24-38).

Figure 4 is an evaluation of the antibody inhibitory activity elicited by a peptide variant of 12 amino acid lengths at the carboxy terminus of hC5a (as shown in SEQ ID NO: 2), and the H of the amino acid position 67 of hC5a is replaced with Amino acid residues of different characteristics (as shown in SEQ ID NOs: 17, 39-55).

Figure 5 is an evaluation of the antibody inhibitory activity elicited by a peptide variant of 12 amino acid lengths at the carboxy terminus of hC5a (as shown in SEQ ID NO: 2), and the M of amino acid position 70 on hC5a is replaced with Different characteristic amino acid residues (as shown in SEQ ID NOs: 18, 56-69).

Figure 6 is an evaluation of the antibody inhibitory activity elicited by a peptide peptide of 12 amino acid lengths at the carboxy terminus of hC5a (as shown in SEQ ID NO: 2), and R of the amino acid position 74 of hC5a and one or The two amino acid positions 67 or 70 are replaced with amino acid residues of different characteristics (as shown in SEQ ID NOs: 70-98).

The above and other objects, features and advantages of the present invention will become more <RTIgt; The term "administration" as used in this context refers to the provision of treatment, for example, the provision of any type of drug or surgical means to a receptor. The treatment is for reversing, slowing, inhibiting development, preventing or reducing the likelihood of disease, disorder, or condition; or for reversing, slowing, inhibiting, or preventing the development, prevention, or reduction of one or more symptoms or diseases, disorders, or conditions. display. The term "prevention" as used in this case is classified as causing a disease, disorder, condition, symptom or manifestation, and does not occur in at least some periods of at least some individuals. "Administration" can include the administration of a medicament to a recipient after one or more symptoms or development of complement regulation, for example, to reverse, slow, reduce severity, and/or inhibit or prevent progression, and/or reversal , slowing, reducing severity, and/or inhibiting the display of one or more symptoms or circumstances. One of the compositions of the present invention can be administered to a receptor that has exhibited a disorder of complement regulation, or a receptor that has a higher development risk (e.g., a disease) than a member of a universal distribution. The composition of the present invention can be administered prophylactically, and typically, prior to the development of any symptoms or environment, the receptor will be at risk in environmental development.

In addition, the recent invention relates to a peptide comprising 7 to 19 amino acid residues, the peptide comprising an amino acid sequence of (X ₃ ) _m KDX ₂ QLGX ₁ (as shown in SEQ ID NO 99), Wherein the X ₁ -monoamine acid residue is selected from the group consisting of alanine, aspartame, glutamine, glycine, histidine, isoleucine, leucine, lysine, methyl sulfide a group consisting of aminic acid, serine, threonine, tyrosine, and valine, the X ₂ -amino acid residue selected from the group consisting of alanine, arginine, histidine, and iso-white the group consisting of alanine, leucine, lysine, methionine, threonine, tyrosine and valine, the amino acid sequence X ₃ is based (X _{4) n} ANISX ₅ ( the SEQ ID NO: 100 as shown), or 1 to 4 comprising the amino acid sequence of amino acid residues of the amino terminal deletion fragment, the amino acid sequence of X ₄ is based VVASQLR (such as SEQ ID NO: 101 Suo An amine-terminal deletion fragment of the amino acid sequence comprising 1 to 6 amino acid residues, the X ₅ -monoamine acid residue selected from the group consisting of alanine, aspartic acid, and glutamine , glutamic acid, histidine, arginine, isoleucine, lysine, methyl sulfide a group consisting of acid, serine, and threonine, wherein m is 0 or 1, and n is 0 or 1, wherein the at least one peptide is coupled or fused to a carrier, and the carrier comprises at least A T cell epitope.

In a preferred embodiment of the invention, the X ₁ -monoamine acid residue is selected from the group consisting of threonine, glutamine, tyrosine, methionine, alanine, glycine, and lysine. The group formed.

When m is 1, X _{1 is} preferably an amino acid residue selected from the group consisting of alanine, aspartame, glutamine, histidine, lysine, methionine, serine and sulphate. a group consisting of amino acids and having an X ₅ -monoamine acid residue selected from the group consisting of alanine, histidine, methionine and threonine, preferably alanine or threonine The acid or methionine, and/or the X ₂ -monoamine acid residue, is selected from the group consisting of methionine, alanine, lysine, and valine.

In a preferred embodiment of the present invention, m is 1, and the X ₅ -monoamine acid residue is selected from the group consisting of alanine, histidine, methionine and threonine, preferably It is alanine, threonine or methionine.

In another preferred embodiment of the invention, X ₂ is an amino acid residue selected from the group consisting of methionine, alanine, lysine, and valine.

In a preferred embodiment of the invention, the peptide is selected from the group consisting of ISHKDMQLGA (shown as SEQ ID NO: 14), ANISHKDMQLGA (shown as SEQ ID NO: 21), and KDMQLGA (shown as SEQ ID NO: 22) ), VVASQLRANISHKDMQLGA (shown as SEQ ID NO: 23), ANISHK DMQLGT (shown as SEQ ID NO: 24), ANISHK DMQLGQ (shown as SEQ ID NO: 25), ANISHK DMQLGY (shown as SEQ ID NO: 26), ANISHKDMQLGM (shown as SEQ ID NO: 27), ANISHK DMQLGG (shown as SEQ ID NO: 28), ANISHK DMQLGV (shown as SEQ ID NO: 29), ANISHKDMQLGK (shown as SEQ ID NO: 30), ANISHKDMQLGS ( As shown in SEQ ID NO: 31, ANISHKDMQLGH (as shown in SEQ ID NO: 32), ANISHK DMQLGN (as shown in SEQ ID NO: 33), AISHIKDMQLGL (as shown in SEQ ID NO: 34), ANISTKDMQLGA (such as SEQ ID NO: 70), ANISTKDMQLGQ (as shown in SEQ ID NO: 71), ANISDKDMQLGS (as shown in SEQ ID NO: 72), ANISTKDMQLGM (such as SEQ) ID NO: 73), ANISMKDMQLGN (shown as SEQ ID NO: 74), ANISTKDKQLGM (as shown in SEQ ID NO: 75), ANISTKDMQLGH (as shown in SEQ ID NO: 76), ANISAKDMQLGA (such as SEQ ID NO: : 77), ANISMKDMQLGA (as shown in SEQ ID NO: 78), ANISTKDKQLGA (as shown in SEQ ID NO: 79), ANISTKDAQLGA (as shown in SEQ ID NO: 80), ANISMKDMQLGS (such as SEQ ID NO: 81) Shown), NISTKDVQLGA (shown as SEQ ID NO: 82), ANISTKDMQLGN (shown as SEQ ID NO: 83), ANISTKDMQLGK (shown as SEQ ID NO: 84), ANISMKDMQLGM (shown as SEQ ID NO: 85) ), ANISTKDMQLGT (shown as SEQ ID NO: 86), ANISHKDKQLGK (shown as SEQ ID NO: 87), ANISMKDMQLGH (shown as SEQ ID NO: 88), and ANISAKDAQLGA (shown as SEQ ID NO: 89) The group that makes up.

In another aspect, the invention relates to a vaccine comprising at least one peptide comprising 7 to 19 amino acid residues, and the amino acid sequence of the peptide is derived from (X ₃ ) _m KDX ₂ QLGX ₁ (shown as SEQ ID NO: 99), wherein the X ₁ is an amino acid residue selected from the group consisting of alanine, aspartame, glutamine, glycine, histidine a group consisting of isoleucine, leucine, lysine, methionine, serine, threonine, tyrosine and valine, preferably arginine, X ₂ An amino acid residue selected from the group consisting of alanine, arginine, histidine, isoleucine, leucine, lysine, methionine, threonine, tyrosine, and valine The group formed is preferably alanine, valine, threonine, tyrosine or leucine, more preferably valine, and the X ₃ is an amino acid sequence (X ₄ ) _n ANISX ₅ (as shown in SEQ ID NO: 100) or an amino acid-terminal deletion fragment of the amino acid sequence comprising 1 to 4 amino acid residues, the X ₄ line being the amino acid sequence VVASQLR (SED ID NO: 101) The amino terminal deletion of the amino acid sequence or the amino acid residue containing 1 to 6 amino acid residues a fragment, the X ₅ -monoamine acid residue selected from the group consisting of alanine, aspartic acid, glutamine, glutamine, histidine, arginine, isoleucine, lysine, a a group consisting of thiaminic acid, serine acid, and threonine, most preferably histidine, wherein m is 0 or 1, and n is 0 or 1, wherein the at least one peptide is a carrier Coupling or fusing, and the vector comprises at least one T cell epitope.

However, another aspect of the invention relates to a vaccine comprising at least one peptide comprising from 7 to 19 amino acid residues, and the amino acid sequence of the peptide is derived from (X ₃ ) _m KDX ₂ QLGX ₁ (shown as SEQ ID NO: 99), wherein the X ₁ is an amino acid residue selected from the group consisting of alanine, aspartic acid, glutamine, glycine, histidine a group consisting of isoleucine, leucine, lysine, methionine, serine, threonine, tyrosine, and valine, preferably arginine, the X ₂ An amino acid residue selected from the group consisting of alanine, arginine, histidine, isoleucine, leucine, lysine, methionine, threonine, tyrosine, and valine the group consisting of, preferably methionine, which is based the amino acid sequence X ₃ (X _{4) n} ANISX ₅ (such as SEQ ID NO: 100 as shown), or comprising amino acid residues from 1 to 4 Based on the amino terminal deletion fragment of the amino acid sequence, the X ₄ line is an amino acid sequence VVASQLR (as shown in SEQ ID NO: 101) or the amino acid sequence comprising 1 to 6 amino acid residues. Amino-terminal deletion fragment, the X ₅ -monoamine acid residue, selected from the group consisting of alanine, day a group consisting of inoguanamine, glutamine, glutamine, arginine, isoleucine, lysine, methionine, serine and threonine, preferably sulphamine Acid, glutamine, glutamic acid, serine, lysine or aspartame, more preferably sulphate or glutamine, wherein m is 0 or 1, and n is 0 or 1 Wherein the at least one peptide is coupled or fused to a carrier and the vector comprises at least one T cell epitope.

In a preferred embodiment of the invention, X ₃ is an amino acid sequence (X ₄ ) _n ANISX ₅ (as shown in SEQ ID NO: 100) or an amine-terminal deletion fragment, and X ₁ is arginine. Thus the amino terminal deletion fragment may comprise ANISX ₅ (as shown in SEQ ID NO: 102), NISX ₅ (as shown in SEQ ID NO: 103), ISX ₅ , SX ₅ or X ₅ ; that is, when m is 1. The vaccine of the present invention may comprise an amino acid sequence of ANISX ₅ KDX ₂ QLGR (as shown in SEQ ID NO 119), NISX ₅ KDX ₂ QLGR (as represented by SEQ ID NO 120), ISX ₅ KDX ₂ QLGR. (As shown in SEQ ID NO 121), at least one peptide of SX ₅ KDX ₂ QLGR (shown as SEQ ID NO 122) or X ₅ KDX ₂ QLGR (as set forth in SEQ ID NO 123).

In a preferred embodiment of the invention, X ₄ is VVASQLR (as set forth in SEQ ID NO: 101) or an amino terminal deletion fragment thereof, and the VVASQLR fragment may be selected from one of the following amino acid sequences: VASQLR ( As shown in SEQ ID NO: 109), ASQLR (as shown in SEQ ID NO: 110), SQLR (as shown in SEQ ID NO: 111), QLR, LR, or R; Yes, when m and n are 1 At least one peptide of the vaccine of the present invention may have one of the following amino acid sequences: VVASQLRANISX ₅ KDX ₂ QLGR (as shown in SEQ ID NO 124), VASQLRANISX ₅ KDX ₂ QLGR (as shown in SEQ ID NO 125) ASQLRANISX ₅ KDX ₂ QLGR (as shown in SEQ ID NO 126), SQLRANISX ₅ KDX ₂ QLGR (as shown in SEQ ID NO 127), QLRANISX ₅ KDX ₂ QLGR (as in SEQ ID NO 128), LRANISX ₅ KDX ₂ QLGR ( As shown in SEQ ID NO 129) or RANISX ₅ KDX ₂ QLGR (as set forth in SEQ ID NO 130).

In a preferred embodiment of the present invention, if X ₂ is an amino acid as defined above and non-methionine, X ₅ in histidines 119 to 130 is histidine.

In a more preferred embodiment of the present invention, if X ₅ is an amino acid as defined above and a non-histidine acid, the X ₂ -methionine in SEQ ID NOs 119-130.

In a preferred embodiment of the invention, the peptide is selected from the group consisting of ANISHK DVQLGR (as set forth in SEQ ID NO 56), ANISHKDTQLGR (shown as SEQ ID NO 57), ANISHKDYQLGR (shown as SEQ ID NO 58), ANISHKDLQLGR (as shown in SEQ ID NO 59), ANISHKDAQLGR (as shown in SEQ ID NO 18), ANISKDMQLGR (as shown in SEQ ID NO 39), ANISQKDMQLGR (as shown in SEQ ID NO 40), ANISEKDMQLGR (such as SEQ ID NO 41) The group shown, ANISSKDMQLGR (shown as SEQ ID NO 42), ANNISKDMQLGR (shown as SEQ ID NO 43), and ANISNKDMQLGR (shown as SEQ ID NO: 44) are preferably selected from the group consisting of A group consisting of ANISHKDVQLGR (shown as SEQ ID NO 56), ANISTKDMQLGR (shown as SEQ ID NO 39) and ANISQKDMQLGR (shown as SEQ ID NO 40).

In another aspect, the invention relates to a peptide selected from the group consisting of ANISHKDVQLGR (as shown in SEQ ID NO 56), ANISHKDTQLGR (shown as SEQ ID NO 57), ANISHKDYQLGR (shown as SEQ ID NO 58), ANISHKDLQLGR (eg, SEQ ID NO 59), ANISHKDAQLGR (shown as SEQ ID NO 18), ANISTKDMQLGR (shown as SEQ ID NO 39), ANISQKDMQLGR (shown as SEQ ID NO 40), ANISEKDMQLGR (shown as SEQ ID NO 41) a group consisting of ANISSKDMQLGR (shown as SEQ ID NO 42), ANESIKDMQLGR (shown as SEQ ID NO 43), and ANISNKDMQLGR (shown as SEQ ID NO: 44), preferably selected from AISHIKDVQLGR (eg SEQ ID) NO 56 shows a group consisting of ANISTKDMQLGR (shown as SEQ ID NO 39) and ANISQKDMQLGR (shown as SEQ ID NO 40).

The object of the present invention is to develop a neutralizing activated immune response against one of the excess human C5a to prevent excessive human C5a from causing pathogenic activity in chronic inflammatory diseases or acute pathological conditions.

To this end, a VARIOTOPE was designed to immunize, thereby eliciting antibodies against new epitopes at the carboxy terminus of human C5a. The new epitope at the carboxy terminus of C5a is cleaved by the C5 protein by the C5 convertase, resulting in a small allergic fragment, C5a and C5b, a member of the membrane attack complex. VARIOTOPE is an immunopeptide that can trigger the humoral immunity of a target protein by calling the antigenic epitope of the target protein. Therefore, the advantage of the VARIOTOPE vaccine is to avoid autotolerance of the autoantigen. In addition, the use of VARIOTOPE can alleviate the unintended side effects caused by the use of the body antigen.

All peptides were chemically linked to the keyhole limpet hemocyanin (KLH) protein carrier via the amino terminus of cysteine and co-administered to mice with aluminum as an adjuvant. Obtained All immune sera from mice immunized with VARIOTOPE or immunized with the original carboxyl sequence of hC5a were analyzed for the antibody titer and functional antibody activity caused by hC5a.

Materials and Methods

Mouse immune response

The BALB/c variety mouse strain was used as an animal model system for the hC5a-VARIOTOPE immune response experiment. BALB/c mothers, 6 to 8 weeks old, were injected with VARIOTOPE (200 μl, subcutaneously injected with a pH=7.4 phosphate solution) in combination with KLH and boosted once every two weeks for 4 times. The aluminum-containing hydrated gel is used as an adjuvant. A total of 5 to 6 mice were used for the VARIOTOPE vaccine for the immune response.

Immunoassay

The immune sera of the vaccine-producing mice were analyzed for their antibody responses by the enzyme-immunized assay (Enzyme Linked Immunosorbent Assay, ELISA) corresponding to the peptides (data not shown) and hC5a protein. The antibody titer is presented as half of the maximum binding amount obtained by serum dilution (for example: ODmax/2) and the average force price of all mice in each group.

C5a inhibition analysis

The inhibitory activity against the C5a antibody elicited by the peptide or VARIOTOPE is carried out by glucuronidase enzyme release assay in human U397 cells. The U397 cell line differentiates with cyclic adenosine 3':5'-monophosphate and stimulates the release of β-glucuronidase by human recombinant C5a protein, which can be blocked by the addition of hC5a-specific antibody or peptide-induced anti-hC5a immune serum. Broken.

In detail, U397 cells were differentiated with cAMP for five days in RPMI containing 10% FBS, and on the fifth day, cells were pretreated with cytochalasin B for ten minutes at 37 °C. Each group of 1.8×10 ⁵ pretreated cells was stimulated with 10 nM hC5a or 8% heat-inactivated serum (56 ° C for one hour) of 10 nM hC5a, wherein the serum was derived from different peptides or VARIOTOPE (see Table 1). And 2 SEQ ID NOs: 1 to 98) immunized mouse serum, and dissolved in a final volume of 120 μl of HAG-CM buffer (containing 20 mM HEPES pH=7.4, 125 mM NaCl, 5 mM KCl, 0.5 mM glucose, 1 mM) CaCl ₂ , 1 mM MgCl ₂ and 0.25% BSA). After stimulation for 10 minutes at 37 ° C, the cells were centrifuged and the supernatant was injected into a 96-well plate with 0.01 M P-nitrophenyl-β-D-glucuronide (dissolved in 0.1 M sodium acetate pH=4.0) at 1: Dilute the ratio of 1 to a total volume of 150 μl. The 96-well plate was incubated at 37 ° C for one hour in the dark, and then 0.4 M glycine buffer (pH = 10.0) was added to stop the reaction. --glucuronidase converts P-nitrophenyl-β-D-glucuronide to yellow and is measured at 405 nm.

Table 1 is the epitope of the carboxy terminus of human C5a and is used as a template for the production of VARIOTOPE.

Table 2 is a VARIOTOPE list of human C5a carboxy-terminal fragments (e.g., SEQ ID NOs: 1-4) in which individual or multiple amino acid groups are replaced with different amino acid residues (in bold).

Table 3 is the abbreviation of all amino acids and their side chain properties.

First Example: Alanine scanning of the hC5a carboxy-terminal epitope (as shown in SEQ ID NOS: 1-4 and Table 1) to define the ability to immunize and induce anti-hC5a antibodies after replacement of the amino acid Maintain even increased amino acid sites.

The individual amino acid groups on the carboxy-terminal epitope of hC5a were replaced with alanine residues to compare the epitope epitope immunity before and after replacement of alanine. All VARIOTOPE can elicit antibodies that bind to the injected peptide, however, the binding cost to the hC5a protein is different. hC5a amino acid 66 (S66A), amino acid 68 (K68A), amino acid 71 (Q71A), amino acid 72 (L72A), amine No. 73, substituted with alanine The base acid (G73A) significantly abolished the production of antibodies recognizing hC5a (as shown in Figures 1A and 1B, SEQ ID NOs: 6, 8, 11, 12, 13, 19 and 20). The original sequence (as shown in SEQ ID NOs: 1-3) elicits relatively high antibody titers, whereas VARIOTOPE (shown as SEQ ID NOs: 6, 8, 11, 12, 13, 19 and 20) elicits antibody potency The price is reduced to less than 13.000 ODmax/2, indicating that the amino acids S, K, Q, L and G (amino acid positions 66, 68, 71, 72 and 73 on hC5a) are important for eliciting hC5a-specific antibodies. (As shown in Figures 1A and 1B, Tables 1-2).

In contrast, the arginine acid at position 74 of the amino acid is replaced by alanine. Significantly increased anti-hC5a antibodies. This is not only shown in the 10 and 12 amino acid lengths of the hC5a carboxy-terminal fragment (as shown in Figures 1A and 1B, SEQ ID NOs: 14, 21), but also in all different lengths of the hC5a carboxy-terminal VARIOTOPE and R74A ( As shown in Figures 1C and 1D, SEQ ID NOs: 22-23). The force range of VARIOTOPE R74A ranged from 56.000 to 88.000 and increased by a factor of 5.5 compared to the original sequence (as shown in Figure 1D, SEQ ID NOs: 4 and 23). VARIOTOPE N64A, I65A, H67A, D69A and M70A (shown as SEQ ID NOs: 5, 7, 9, 10, 15-18) exhibit anti-hC5a forces similar to the original epitope SEQ ID NOs: 1-2 Price (as shown in Figures 1A and 1B, Tables 1 and 2). In summary, substitution of a single amino acid on a different length of the hC5a carboxy-terminal epitope resulted in similar results, indicating that the specific amino acid residue is only slightly affected by the peptide length for hC5a immunity. In particular, the replacement of arginine at position 74 of the amino acid position on the carboxy terminal fragment of hC5a with alanine resulted in a significant increase in the potency of the anti-hC5a after replacement compared to the original epitope. The peptide fragment used to make the vaccine response contains at least the last 7 amino acids of the carboxy terminus of hC5a to ensure immunity, and may not exceed 19 amino acids, which is the defined length of the new epitope of the carboxy-terminal end of hC5a.

The inhibitory activity of VARIOTOPE in which each amino acid was replaced with alanine was evaluated by glucuronidase enzyme release assay. Briefly, differentiated human U937 cells are stimulated by hC5a to release beta-glucuronidase, which is blocked by anti-hC5a immune sera.

The immune sera elicited by VARIOTOPE R74A showed the best inhibitory capacity, and its inhibitory activity was increased by a factor of two relative to the original sequence (as shown in SEQ ID NOs: 1-4) (eg, Figure 2A-D, SEQ ID NOs: 14). , 21, 22 and 23). Furthermore, the immune sera elicited by SEQ ID NOs: 5, 7, 10, 17 and 18 have an increased or decreased inhibitory activity compared to the original epitope (as shown in Figures 2A and 2B). Thus, H67A (as shown in SEQ ID NOs: 7 and 17), M70A (as shown in SEQ ID NOs: 10 and 18) and 10 amino acid lengths of I65A (eg SEQ ID NO: 5; instead of 12) Amino acid length I65A, SEQ ID NO: 16) appears to be beneficial for the induction of functional anti-hC5a antibodies; however, R74A has advantages in VARIOTOPE of different lengths (as shown in Figures 2A to D).

There is a good correlation between the protein valence of anti-hC5a and the functional activity measured by glucuronidase release assay. In the subsequent examples, only the immune serum (antibody) inhibitory activity elicited by the original antigenic epitope and VARIOTOPE were revealed, and in principle, the efficiency was more predictive than the antibody titer alone.

SECOND EMBODIMENT: The hC5a important amino acid defined by alanine scanning is the arginine acid at the amino acid position 74, which causes more than twice the inhibitory activity of hC5a (as shown in Figure 2). . Therefore, in the next experiment, the amino acid No. 74 was systematically substituted into a variety of amino acids with smaller or opposite characteristic residues relative to arginine residues (as shown in Table 3). ). In this experiment, a carboxy-terminal epitope with 12 amino acid lengths was chosen as the template because of the fragmentation compared to 10 or 20 amino acid lengths (as shown in Figures 1A and D). A fragment of amino acid length (as shown in Figure 1B) elicits a higher anti-hC5a antibody titer and the 12 amino acid length fragments compared to the seven amino acid length hC5a carboxyl end fragments Preferred immunological activity is also exhibited.

Sixteen R74X VARIOTOPEs with 12 amino acid-length hC5a carboxy-terminal epitopes were tested for their immunogenicity and their ability to elicit functionally active antibodies. The immune sera obtained from R74T and R74Q VARIOTOPE showed the best inhibitory effect on glucuronidase release assay (as shown in Figure 3, SEQ ID NOs: 24 and 25), and the suboptimal effect was R74Y (as in Figure 3, SEQ ID NO: 26) and replacement of R with non-polar aliphatic amino acid residues M, A, G and V (as shown in Figure 3, SEQ ID NOs: 27, 21, 28 and 29). When R in VARIOTOPE is replaced by a negatively charged amino acid (R74D) or an aromatic, non-polar amino acid (W and F) and an amino acid that affects the structure, such as P, it does not tend to induce hC5a inhibition. Sex antibodies (as shown in Figure 4, SEQ ID NOs: 35-38).

Third embodiment: histidine acid at position 67 of hC5a, which is C Another amino acid that tends to be replaced in the amino acid scan. Thus, the amino acid No. 67 was also systematically substituted into a plurality of amino acids having smaller or opposite characteristic residues relative to histamine residues (as shown in Table 3). Eighteen H67X VARIOTOPEs with 12 amino acid-length hC5a carboxy-terminal epitopes were tested for their immunogenicity and their ability to elicit functionally active antibodies (as shown in SEQ ID NOs: 17, 39-55).

Immune sera obtained from H67T and H67Q VARIOTOPE showed the best inhibitory effect on glucuronidase release assay (as shown in Figure 4, SEQ ID NOs: 39 and 40), and its inhibitory effect was greater than the original sequence (eg SEQ ID) NO: 2) increases by 20% (as shown in Figure 4). The inhibitory effect of immune sera obtained from SEQ ID NOs: 41-47 VARIOTOPE showed a slight increase or the same as the original sequence (as shown in SEQ ID NO: 2). SEQ ID NOs: 51-55 VARIOTOPE-induced immune sera caused a significant decrease in hC5a inhibition, which was reduced by more than 20% compared to the original sequence (as shown in SEQ ID NO: 2) (as shown in Figure 4).

The fourth embodiment: the methionine at position 70 of the amino acid on hC5a is the next systemic replacement to obtain a higher carboxy terminal epitope than the original 12 amino acids of hC5a. VARIOTOPE against hC5a inhibition. Fifteen VARIOTOPEs were analyzed for glucuronidase release to test their functional activity. The VARIOTOPE-primed immune sera of SEQ ID NOs: 18 and 56-62 showed similar or similar inhibitory effects as the original antigenic epitope (as shown in SEQ ID NO: 2) (as shown in Figure 5). However, VARIOTOPE of SEQ ID NOs: 63-60 shows a progressively decreasing inhibitory activity and does not tend to elicit a functionally active antibody against hC5a.

Fifth Example: The position of the amino acid No. 74 on hC5a has an important effect on the immunological ability of the carboxy-terminal fragment of hC5a, and also affects the functional activity of the antibody elicited by it (as shown in Fig. 3). However, such an effect is substantiated after the substitution of the 67th or the 70th of the hC5a carboxy terminal epitope or the simultaneous replacement of the two amino acid positions. Including R74X In the experiment of VARIOTOPE with another amino acid substitution position 67 or 70, or the simultaneous replacement of two amino acid positions, the immunity was tested separately. H67T, H67M and H67A are replaced by non-polar small amino acid residues (A, M), polar uncharged amino acid residues (Q, S, N) and positively charged with the amino acid position R of the 74th. After the amino acid residue (H), a higher anti-hC5a antibody titer and a nearly 1.5-fold increase in active antibody than the original antigen epitope (as shown in SEQ ID NO: 2) are obtained (eg, Figure 6, SEQ) ID NOs: 70~80). A favorable substitution of the position of the amino acid No. 74, with the positional replacement of the amino acid No. 67 as H67T, H67M and H67A, or with the positional replacement of the amino acid No. 7 as M70K, M70A and M70V, may cause a ratio The original sequence (as shown in SEQ ID NO: 2) has a higher inhibitory activity. Compared to the original sequence, VARIOTOPE SEQ ID NOs: 90-98 elicited immune serum to reduce inhibitory activity (as shown in Figure 6).

While the invention has been described in connection with the preferred embodiments described above, it is not intended to limit the scope of the invention. The technical scope of the invention is protected, and therefore the scope of the invention is defined by the scope of the appended claims.

<110> Affiris AG

<120> Composition (Composition)

<130> R 63571

<150> EP 12166314.0

<151> 2012-05-01

<160> 196

<170> PatentIn version 3.5

<210> 1

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 1

<210> 2

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 2

<210> 3

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 3

<210> 4

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 4

<210> 5

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 5

<210> 6

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 6

<210> 7

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 7

<210> 8

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 8

<210> 9

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 9

<210> 10

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 10

<210> 11

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 11

<210> 12

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 12

<210> 13

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 13

<210> 14

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 14

<210> 15

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 15

<210> 16

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 16

<210> 17

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 17

<210> 18

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 18

<210> 19

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 19

<210> 20

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 20

<210> 21

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 21

<210> 22

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 22

<210> 23

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 23

<210> 24

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 24

<210> 25

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 25

<210> 26

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 26

<210> 27

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 27

<210> 28

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 28

<210> 29

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 29

<210> 30

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 30

<210> 31

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 31

<210> 32

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 32

<210> 33

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 33

<210> 34

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 34

<210> 35

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 35

<210> 36

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 36

<210> 37

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 37

<210> 38

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 38

<210> 39

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 39

<210> 40

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 40

<210> 41

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 41

<210> 42

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 42

<210> 43

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 43

<210> 44

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 44

<210> 45

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 45

<210> 46

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 46

<210> 47

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 47

<210> 48

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 48

<210> 49

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 49

<210> 50

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 50

<210> 51

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 51

<210> 52

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 52

<210> 53

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 53

<210> 54

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 54

<210> 55

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 55

<210> 56

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Artificial peptide sequence

<400> 56

<210> 57

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> Common sequence

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa = (any amino acid residue) n, wherein n = 0 or an integer greater than 0

<220>

<221> MISC_FEATURE

<222> (2)..(2)

<223> Xaa = monoamino acid residue is selected from the group consisting of aspartic acid (D) and glutamic acid (glutamic acid, E) group

<220>

<221> MISC_FEATURE

<222> (3)..(3)

<223> Xaa = any amino acid residue

<220>

<221> MISC_FEATURE

<222> (6)..(6)

<223> Xaa = any amino acid residue

<220>

<221> MISC_FEATURE

<222> (7)..(7)

<223> Xaa=monoamino acid residue is selected from the group consisting of proline (P) and alanine (A)

<220>

<221> MISC_FEATURE

<222> (8)..(8)

<223> Xaa = monoamino acid residue is selected from the group consisting of aspartic acid (D) and glutamic acid (glutamic acid, E) group

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa = (any amino acid residue) n, wherein n = 0 or an integer greater than 0

<400> 57

<210> 58

<211> 140

<212> PRT

<213> Human

<400> 58

<210> 59

<211> 134

<212> PRT

<213> Human

<400> 59

<210> 60

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> Human A-type nuclear protein fragment

<400> 60

<210> 61

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 61

<210> 62

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 62

<210> 63

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 63

<210> 64

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 64

<210> 65

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 65

<210> 66

<211> 6

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 66

<210> 67

<211> 10

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 67

<210> 68

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 68

<210> 69

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 69

<210> 70

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 70

<210> 71

<211> 6

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 71

<210> 72

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 72

<210> 73

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 73

<210> 74

<211> 11

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 74

<210> 75

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 75

<210> 76

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 76

<210> 77

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 77

<210> 78

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 78

<210> 79

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 79

<210> 80

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 80

<210> 81

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 81

<210> 82

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 82

<210> 83

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 83

<210> 84

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 84

<210> 85

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 85

<210> 86

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 86

<210> 87

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 87

<210> 88

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 88

<210> 89

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 89

<210> 90

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 90

<210> 91

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 91

<210> 92

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 92

<210> 93

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 93

<210> 94

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 94

<210> 95

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 95

<210> 96

<211> 13

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 96

<210> 97

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 97

<210> 98

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 98

<210> 99

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 99

<210> 100

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 100

<210> 101

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 101

<210> 102

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 102

<210> 103

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 103

<210> 104

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 104

<210> 105

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 105

<210> 106

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 106

<210> 107

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 107

<210> 108

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 108

<210> 109

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 109

<210> 110

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 110

<210> 111

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 111

<210> 112

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 112

<210> 113

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 113

<210> 114

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 114

<210> 115

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 115

<210> 116

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 116

<210> 117

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 117

<210> 118

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 118

<210> 119

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 119

<210> 120

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 120

<210> 121

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 121

<210> 122

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 122

<210> 123

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 123

<210> 124

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 124

<210> 125

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 125

<210> 126

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 126

<210> 127

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 127

<210> 128

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 128

<210> 129

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 129

<210> 130

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 130

<210> 131

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 131

<210> 132

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 132

<210> 133

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 133

<210> 134

<211> 8

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<400> 134

<210> 135

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (2)..(2)

<223> Xaa is an amino acid residue selected from the group consisting of lysine (K) and arginine (R). a group consisting of alanine (A) and histidine (H)

<220>

<221> MISC_FEATURE

<222> (3)..(3)

<223> Xaa is an amino acid residue selected from asparagine (N), glutamic acid (glutamine, Q), serine (S), glycine (Glycine, G) and Group of alanine (A)

<220>

<221> MISC_FEATURE

<222> (4)..(4)

<223> Xaa is an amino acid residue selected from Glutamic acid (E), aspartic acid (D) and alanine Group of (alanine, A)

<220>

<221> MISC_FEATURE

<222> (5)..(5)

<223> Xaa is an amino acid residue selected from the group consisting of glutamic acid (E) and aspartic acid Group of (aspartic acid, D)

<220>

<221> MISC_FEATURE

<222> (7)..(7)

<223> Xaa is an amino acid residue selected from Group of alanine (A) and tyrosine (Y)

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 135

<210> 136

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 136

<210> 137

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 137

<210> 138

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 138

<210> 139

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 139

<210> 140

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 140

<210> 141

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 141

<210> 142

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 142

<210> 143

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 143

<210> 144

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 144

<210> 145

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 145

<210> 146

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 146

<210> 147

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 147

<210> 148

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 148

<210> 149

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 149

<210> 150

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 150

<210> 151

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 151

<210> 152

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 152

<210> 153

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 153

<210> 154

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 154

<210> 155

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cystine, and n is 0 or an integer of more than 0, preferably 1 Or 0

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine,and m is 0 or an integer of more than 0,preferably 1 Or 0

<400> 155

<210> 156

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 156

<210> 157

<211> 14

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> misc_feature

<222> (14)..(14)

<223> Xaa can be any naturally occurring amino acid

<400> 157

<210> 158

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 158

<210> 159

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 159

<210> 160

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 160

<210> 161

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 161

<210> 162

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 162

<210> 163

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 163

<210> 164

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 164

<210> 165

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 165

<210> 166

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)m, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 166

<210> 167

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 167

<210> 168

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 168

<210> 169

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 169

<210> 170

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 170

<210> 171

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 171

<210> 172

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 172

<210> 173

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 173

<210> 174

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 174

<210> 175

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 175

<210> 176

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 176

<210> 177

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 177

<210> 178

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 178

<210> 179

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 179

<210> 180

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 180

<210> 181

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 181

<210> 182

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 182

<210> 183

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 183

<210> 184

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 184

<210> 185

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 185

<210> 186

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 186

<210> 187

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 187

<210> 188

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 188

<210> 189

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 189

<210> 190

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 190

<210> 191

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400>191

<210> 192

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 192

<210> 193

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 193

<210> 194

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 194

<210> 195

<211> 9

<212> PRT

<213> Artificial sequence

<220>

<223> pseudo antigenic epitope

<220>

<221> MISC_FEATURE

<222> (1)..(1)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<220>

<221> MISC_FEATURE

<222> (9)..(9)

<223> Xaa=(Xaa)n, wherein Xaa is any amino acid residue, preferably Cysteine, and n is 0 or an integer greater than 0, preferably 1 or 0.

<400> 195

<210> 196

<211> 7

<212> PRT

<213> Artificial sequence

<220>

<223> Human A-type nuclear protein fragment

<400> 196

Claims (14)

A vaccine comprising at least one peptide comprising 7 to 19 amino acid residues, and the amino acid sequence of the peptide (X ₃ ) _m KDX ₂ QLGX ₁ (eg SEQ ID NO: 99) Shown) wherein the X ₁ -amino acid residue is selected from the group consisting of alanine, aspartame, glutamine, glycine, histidine, isoleucine, leucine, lysine a group consisting of aminic acid, methionine, serine, threonine, tyrosine, and valine, the X ₂ -amino acid residue selected from the group consisting of alanine, arginine, and a group the group consisting of leucine, isoleucine, leucine, lysine, methionine, threonine, tyrosine and valine, the amino acid sequence X ₃ is based (X _{4 n} ANISX ₅ (as shown in SEQ ID NO: 100) or an amino terminal deletion fragment of the amino acid sequence comprising 1 to 4 amino acid residues, the X ₄ line being the amino acid sequence VVASQLR (eg SEQ) ID NO: 101) or an amino terminal deletion fragment of the amino acid sequence comprising 1 to 6 amino acid residues, the X ₅ -monoamine acid residue selected from the group consisting of alanine and aspartic acid , glutamine, glutamic acid, histidine, arginine, isoleucine, lysine a group consisting of methionine, serine, and threonine, wherein m is 0 or 1, and n is 0 or 1, wherein the at least one peptide is coupled or fused to a carrier, and The vector comprises at least one T cell epitope. The vaccine according to claim 1, wherein the X ₁ -amino acid residue is selected from the group consisting of sulphate, glutamine, tyrosine, methionine, alanine, and glycine. a group consisting of acid and proline. The vaccine according to claim 1, wherein the X ₁ -amino acid residue is selected from the group consisting of alanine, aspartame, glutamine, histidine, lysine, and methyl sulfide. a group consisting of aminic acid, serine and threonine, m is 1, the X ₅ -monoamine acid residue, selected from the group consisting of alanine, histidine, methionine and threonine a group, preferably alanine, threonine or methionine, and/or X ₂ is an amino acid residue selected from the group consisting of methionine, alanine, lysine and proline. The group that makes up. The vaccine according to claim 1 or 2, wherein m is 1, and X ₅ is an amino acid residue selected from the group consisting of methionine, alanine, lysine and valine. Group. The vaccine of claim 1, wherein the X ₂ -monoamine acid residue is selected from the group consisting of methionine, alanine, lysine, and valine. group. The vaccine according to any one of claims 1 to 5, wherein the at least one peptide is selected from the group consisting of ISHKDMQLGA (as shown in SEQ ID NO: 14) and ANISHKDMQLGA (as shown in SEQ ID NO: 21). ), KDMQLGA (shown as SEQ ID NO: 22), VVASQLRANISHKDMQLGA (shown as SEQ ID NO: 23), ANISHK DMQLGT (shown as SEQ ID NO: 24), ANISHK DMQLGQ (shown as SEQ ID NO: 25), ANISHKDMQLGY (shown as SEQ ID NO: 26), ANISHK DMQLGM (shown as SEQ ID NO: 27), ANISHKDMQLGG (shown as SEQ ID NO: 28), ANISHK DMQLGV (shown as SEQ ID NO: 29), ANISHKDMQLGK ( As shown in SEQ ID NO: 30, ANISHKDMQLGS (as shown in SEQ ID NO: 31), ANISHKDMQLGH (as shown in SEQ ID NO: 32), ANISHKDMQLGN (as shown in SEQ ID NO: 33), ANISHKDMQLGL (such as SEQ ID NO: 34), ANISTKDMQLGA (as shown in SEQ ID NO: 70), ANISTKDMQLGQ (as shown in SEQ ID NO: 71), ANISDKDMQLGS (as shown in SEQ ID NO: 72), ANISTKDMQLGM (such as SEQ ID NO: :73), ANISMKDMQLGN (as shown in SEQ ID NO: 74), ANISDKDKQLGM (as shown in SEQ ID NO: 75), ANISDKDMQLGH (as shown in SEQ ID NO: 76), ANISAKDMQLGA (as shown in SEQ ID NO: 77), ANISMKDMQLGA (as shown in SEQ ID NO: 78), ANISTKDKQLGA (as shown in SEQ ID NO: 79), ANISTKDAQLGA (as shown in SEQ ID NO: 80), ANISMKDMQLGS (eg SEQ ID NO: 81), NISTKDVQLGA (shown as SEQ ID NO: 82), ANISTKDMQLGN (shown as SEQ ID NO: 83), ANISTKDMQLGK (shown as SEQ ID NO: 84), ANISMKDMQLGM (eg SEQ ID) NO: 85), ANISTKDMQLGT (shown as SEQ ID NO: 86), ANISHKDKQLGK (shown as SEQ ID NO: 87), ANISMKDMQLGH (shown as SEQ ID NO: 88), and ANISAKDAQLGA (eg SEQ ID NO: The group consisting of 89). The vaccine according to any one of claims 1 to 6, wherein the at least one peptide comprises at least one half of a cysteine residue at the amino terminus, and the at least one cysteine residue is directly or The amino terminus of the peptide is bound in a spacer sequence. The vaccine according to any one of claims 1 to 7, wherein the vector comprising at least one T cell epitope is a protein carrier. The vaccine according to claim 8, wherein the protein carrier is selected from the group consisting of keyhole limpet hemocyanin (KLH), CRM197, tetanus toxoid (TT), protein D or diphtheria toxin (DT). The vaccine according to any one of claims 1 to 9, wherein the compound is complexed with an adjuvant, preferably with aluminum. The vaccine according to any one of claims 1 to 10, which is for treating and/or preventing a disease associated with a complement system. The vaccine according to claim 11, wherein the complement system-related disease is an inflammatory disease, preferably a chronic inflammatory disease. The vaccine according to claim 12, wherein the inflammatory disease is selected from the group consisting of Annual macular degeneration (AMD), neurodegenerative diseases, especially Alzheimer's disease, Parkinson's disease or Huntington's disease, asthma, atherosclerosis, vasculitis, dermatitis, especially sputum And a group consisting of urticaria, hemolytic uremic syndrome, rheumatoid arthritis, Guillain-Barré syndrome, multiple sclerosis, antiphospholipid antibody syndrome, hemolytic uremic syndrome, and lupus erythematosus (SLE). The vaccine according to claim 11, wherein the complement system related diseases are selected from the group consisting of ischemia-reperfusion injury, acute lung injury, acute respiratory distress syndrome, sepsis, cancer, pregnancy complications, such as pregnancy hypertension , a group consisting of spontaneous abortion, growth retardation, and hemodialysis-related thrombosis.