CN103547283A

CN103547283A - HLA-binding peptides derived from prostate-associated antigenic molecules and methods of use thereof

Info

Publication number: CN103547283A
Application number: CN201180059274.2A
Authority: CN
Inventors: 托尼·怀申克; 彼得·莱维卓尔斯奇; 汉斯—宙戈·冉曼思; 斯蒂芬·斯蒂万诺维奇; 赛瑟利·戈泰芬奇斯
Original assignee: Immatics Biotechnologies GmbH
Current assignee: Immatics Biotechnologies GmbH
Priority date: 2010-12-14
Filing date: 2011-11-14
Publication date: 2014-01-29
Also published as: MX2013006758A; KR20130126671A; JP6032853B2; WO2012079878A2; AU2011344652A1; NZ609916A; CA2821582A1; JP2014502961A; WO2012079878A3; SG191154A1; AU2011344652B2

Abstract

Methods and compositions for immunotherapeutic treatment of prostate cancer are disclosed. More specifically methods of treating patients with prostate cancer comprising administering compositions comprising HLA-binding peptides derived from prostate-associated antigenic molecules, either with or without immunological adjuvants, are disclosed.

Description

Human leukocyte antigen's binding peptide and the using method thereof in prostate related antigen molecule source

Invention field

The invention discloses the method and composition of carcinoma of prostate immunization therapy.More particularly, disclose the Therapeutic Method of patients with prostate cancer, comprised and combine the medication of using or using separately containing the compositions of prostate related antigen molecule source human leukocyte antigen (HLA) binding peptide with immunological adjuvant.

Background

Carcinoma of prostate is one of common cancer of male, and the number of the infected of Europe report in 2006 is 346,000 people, and death toll is about 87,000 people.In the U.S., carcinoma of prostate is the modal cancer diagnosis of male, is also the second largest reason of cancer associated death.Because the sensitivity of prostate specific antigen (Prostate specific antigen, PSA) monitoring analysis improves, carcinoma of prostate can detect with the clinical focal stage in early days, can use radical treatment to treat as operation and radioactive method.Yet these patients had the probability of 10 – 60% to there will be that PSA is asymptomatic to be increased in 10 years, be called " biochemical recurrence ".Biochemical recurrence typically refers to hiding localized cancer recurrence or the morbidity of metastasis not yet.

In this case, Therapeutic Method has external exposure and androgen-deprivation (removing also referred to as hormone).But two kinds of methods are not all proved to be effectively, particularly to extend patient's long-term surviving time aspect.And, these two kinds of Therapeutic Method are restricted because of multiple side effect, side effect comprises cardiovascular problems, osteoporosis, body weight increase, neuro-cognitive decline, forms urethral stricture, hyposexuality and the increase of sexual impotence risk, and risk and pathologisch Bruch risk that osteoporosis skeleton calcium salt reduces significantly increase.In addition, someone worries that androgen-deprivation can allow androgen independence tumor be cloned in early origin, finally causes long-term tumour progression further to be accelerated.And there is dispute the best opportunity that starts androgen-deprivation treatment, be particularly characterized as the biochemistry recurrence of low PSA value or long mean doubling time (DT).In view of these risks, androgen-deprivation treatment and external exposure are to all ambigendi locus of the therapeutic value of early stage biochemical patients with recurrent.

Tumor associated antigen (" TAA ") has been confirmed as potential Immunotherapeutic agent for cancer.Determine various dissimilar tumors and tissue-specific a large amount of TAA, comprised the TAA that those are relevant to prostate.TAA specific T-cells in cancer patient's prostate tumor tissue, tumor-draining lymphode and around blood circulation identify and immunization therapy after specific T-cells reaction increase and all show, use TAA to control immune system and may contribute to treat carcinoma of prostate.

The various systems of prostate specific TAA body endoantigen having been offered in clinical trial are checked, and comprising: (1) carries out vaccine therapy with autologous or allogeneic tumor cell; (2) autologous tumor cell of construction expression granulocyte-macrophage colony stimutaing factor; (3) with HLA I and HLA II binding peptide, dendritic cell is carried out to external impulse; (4) dendritic cell of tumor-mRNA transfection; (5) express the vaccinia virus recombinant of TAA.But these researchs are most to be carried out in androgen resistance type patients with prostate cancer.The information of the androgen responsive type patient vaccine therapy recurring about the front chemistry of androgen-deprivation treatment seldom.

Therefore, patients with prostate cancer, particularly early stage biochemical patients with recurrent need to have new Therapeutic Method.

Summary

The disclosure relates to the compoistion and method of use for immunization therapy.Particularly, the disclosure relates to cancer, and especially carcinoma of prostate, more specifically says the immunization therapy of the androgen responsive type carcinoma of prostate of not accepting androgen-deprivation treatment, early stage chemical patients with recurrent.The disclosure also relates to the compositions of HLA I class or II class HLA binding peptide, described HLA binding peptide derives from prostate related antigen molecule, as prostate specific antigen, prostate stem cell antigen, prostate specific membrane antigen, survivin, prostate specific albumen and transient receptor potential-p8(" TRP-p8 ").

On the one hand, the disclosure relates to the compositions that comprises at least one HLA binding peptide, and wherein said HLA binding peptide comprises the epi-position from prostate related antigen molecule.

On the other hand, the disclosure relates to the compositions that comprises at least one HLA binding peptide, and wherein said HLA binding peptide comprises to be freely selected from the epi-position with the prostate related antigen molecule in next group antigen: prostate specific antigen, prostate stem cell antigen, prostate specific membrane antigen, survivin, prostate specific albumen and transient receptor potential-p8.

The present invention particularly preferably aspect relates to the compositions that comprises at least two kinds of HLA binding peptides, wherein: (a) at least two kinds of HLA binding peptides, have at least one peptide to contain sequence numbering: 23 epi-position or sequence numbering: 23 fusion rotein, 80 aminoacid that this fusion rotein contains the relevant invariant chain N end of HLA-DR antigen.(b) at least two kinds of peptides, there is at least one peptide containing the epi-position of selecting from following one group, comprising: sequence numbering: 1 to 11,13 to 22,24 to 42.

Preferable case is that compositions of the present invention comprises at least two kinds by b) peptide that forms of group aminoacid sequence.

Preferred composition is compositions of the present invention, wherein said compositions comprises at least two kinds of peptides, be preferably at least 4 kinds of peptides, 10 kinds of peptides more preferably, by sequence numbering: 23 aminoacid sequence forms, a kind of peptide is to select from following one group of sequence numbering: sequence numbering: 1,2,5,7,9 to 11,13 and 14, and at least one peptide is to select from following one group of sequence numbering: sequence numbering: 15 to 22,24 to 42.

Equally, preferred composition is compositions of the present invention, and wherein, extra peptide is to select according to experimenter HLA group to be treated.

Preferred composition is compositions of the present invention, wherein said compositions comprises at least 4 kinds by sequence numbering: the peptide that 23 aminoacid sequence forms, a kind of peptide is to select from following one group of sequence numbering: sequence numbering: 1,2,5,7,9 to 11,13 and 14, and at least one peptide is to select from following one group of sequence numbering: sequence numbering: 15 to 22 and sequence numbering: 24 to 42.

Meanwhile, preferred composition is compositions of the present invention, and wherein, at least one peptide is II class peptide.

So, relate on the other hand compositions of the present invention, further comprise the mixture of immunological adjuvant or two or three immunological adjuvant, such as GMCSF and imiquimod.

Preferred composition is the true compositions of the present invention, and wherein, this immunological adjuvant contains Toll sample receptor stimulating agent, as Toll sample receptor-7 agonist.

Meanwhile, preferred composition is compositions of the present invention, contains at least one antigen presenting cell, and dendritic cell for example, just like the autologous dendritic cell of peptide impulse or peptide load.

Relate to so, on the other hand the application of the present composition in prostate cancer therapy.

Preferred composition is compositions of the present invention, wherein, described carcinoma of prostate be androgen responsive type and patient do not accept androgen-deprivation treatment.

Further preferred compositions is compositions of the present invention, and wherein, described carcinoma of prostate is androgen insensitivity type.

So, relate on the other hand the method for the treatment of carcinoma of prostate, comprise the present composition that gives patient's effective dose.

Method for optimizing is method of the present invention, wherein, described carcinoma of prostate be androgen responsive type and patient do not accept androgen-deprivation treatment.

Further preferred method is method of the present invention, and wherein, described carcinoma of prostate is androgen insensitivity type.

On the other hand, the epi-position that compositions of the present disclosure comprises following sequence numbering: sequence numbering: 24,7,1,2,3,4,5,6,8,9,10,11,13,14 or sequence numbering: 15 to 23 or sequence numbering: 25 to 40.

On the other hand, compositions at least comprises two kinds of HLA binding peptides that contain prostate related antigen molecule source epi-position, and wherein at least two kinds of HLA binding peptides, having at least one is HLA I class peptide; In at least two kinds of HLA binding peptides, having at least one is HLA II class peptide.

The compositions that comprises at least two kinds of HLA binding peptides that relates in one aspect to again of the present disclosure, wherein in HLA binding peptide, has a kind of HLA of being I class peptide at least, it contains the epi-position of selecting from following a group: sequence numbering: 1,2,3,4,5,6,7,8,9,10 and 11, in HLA binding peptide, have a kind of HLA of being II class peptide at least, it contains the epi-position of selecting from following a group: sequence numbering: 13 and 14.

The compositions that comprises at least two kinds of HLA binding peptides that relates in one aspect to again of the present disclosure, wherein has at least a kind of peptide to contain the epi-position of selecting from following one group in (a) at least two kinds of HLA binding peptides: sequence numbering: 24, sequence numbering: 15 to 23 and sequence numbering: 25 to 37; (b) at least two kinds of HLA binding peptides, have at least a kind of peptide to contain the epi-position of selecting from following one group: sequence numbering: 7,1 to 6,8 to 11,13,14, and sequence numbering: 38 to 42.

The compositions that comprises at least two kinds of HLA binding peptides that also relates on the one hand more of the present disclosure, wherein in HLA binding peptide, have a kind of HLA of being I class peptide at least, it mainly contains the epi-position of selecting from following a group: sequence numbering: 1,2,3,4,5,6,7,8,9,10 and 11; In HLA binding peptide, have a kind of HLA of being II class peptide at least, it mainly contains the epi-position of selecting from following a group: sequence numbering: 13 and 14.

The compositions that comprises at least two kinds of HLA binding peptides that also relates on the one hand more of the present disclosure, wherein in HLA binding peptide, have a kind of HLA of being I class peptide at least, it contains the epi-position of selecting from following a group: sequence numbering: 1,2,3,4,5,6,7,8,9,10 and 11; In HLA binding peptide, have a kind of HLA of being II class peptide at least, it contains the epi-position of selecting from following a group: sequence numbering: 13 and 14.

Of the present disclosurely on the one hand also relate to the compositions that comprises at least two kinds of HLA binding peptides again, wherein in HLA binding peptide, having at least a kind of is with allele but not HLA I class peptide that HLA-A*02 is combined.

The compositions that comprises at least two kinds of HLA binding peptides that relates in one aspect to again of the present disclosure, wherein in HLA binding peptide, having at least a kind of is the HLA I class peptide of being combined with the allele that is selected from mono-group of HLA-A*24, HLA-A*11, HLA-B*41, HLA-B*51 or HLA-C.

One side more of the present disclosure also relates to the compositions that comprises at least 6 kinds of HLA binding peptides, wherein: (a) at least one HLA binding peptide contains the epi-position from prostate specific antigen; (b) at least one HLA binding peptide contains the epi-position from prostate stem cell antigen; (c) at least one HLA binding peptide contains the epi-position from prostate specific membrane antigen; (d) at least one HLA binding peptide contains the epi-position from survivin; (e) at least one HLA binding peptide contains the epi-position from prostate specific albumen; (f) at least one HLA binding peptide contains the epi-position from transient receptor potential-p8.

Another aspect of the present disclosure relates to above-mentioned composition, and wherein said composition comprises at least two kinds by b) peptide that forms of group aminoacid sequence.

Another aspect of the present disclosure relates to the above-mentioned composition of the HLA binding peptide that comprises following sequence numbering: sequence numbering: 1,2,3,4,5,6,7,8,9,10,11,13,14,15,16,17,18,19,20,21,22,23,24,25,26,27,28,29,30,31,32,33,34,35,36,37,38,39,40,41 and 42.

Another aspect of the present disclosure relates to above-mentioned composition, and wherein said compositions comprises the peptide that the aminoacid sequence by following sequence numbering forms: sequence numbering: 1, sequence numbering: 2; Sequence numbering: 5, sequence numbering: 7, sequence numbering: 9 to sequence numbering: 11, sequence numbering: 13 and sequence numbering: 14, and at least one peptide is to select (optional) from following one group of sequence numbering: 15 to sequence numbering: 42.

Another aspect of the present disclosure relates to a kind of above-mentioned compositions, wherein said compositions comprises at least two kinds of peptides, at least 4 kinds of peptides preferably, be more preferably 10 kinds of peptides, aminoacid sequence by following sequence numbering forms: sequence numbering: 1,2,5,7,9 to 11,13 and 14, and at least one peptide is to select from following one group of sequence numbering: sequence numbering: 15 to 42.

Another aspect of the present disclosure relates to a kind of above-mentioned compositions, wherein said compositions comprises at least two kinds of peptides, at least 4 kinds of peptides preferably, be more preferably 10 kinds of peptides, aminoacid sequence by following sequence numbering forms: sequence numbering: 1,2,5,7,9 to 11,13 and 14, at least one peptide is to select from following one group of sequence numbering: sequence numbering: 15 to 42, and wherein, extra peptide is to select according to experimenter HLA group to be treated.

So, another aspect of the present disclosure relates to above-mentioned composition, and wherein, described compositions comprises at least 4 kinds of peptides that are comprised of the aminoacid sequence of following sequence numbering: sequence numbering: 1,2,5,7, sequence numbering: 9 to 11, sequence numbering: 13,14 and sequence numbering: 15 to 42.

Another aspect of the present disclosure relates to above-mentioned any compositions, and wherein, at least one peptide is II class peptide.

Another aspect of the present disclosure relates to above-mentioned any compositions, further comprises the mixture of immunological adjuvant or two or three immunological adjuvant, such as GMCSF and imiquimod.Immunological adjuvant can be any known immunological adjuvant, comprises Toll sample receptor-7 agonist imiquimod and Mucin1-mRNA/ protamine complex, and cytokine-granulocyte-macrophage colony stimutaing factor (GM-CSF).

So, another aspect of the present disclosure relates to the method for the treatment of carcinoma of prostate, comprises combining with immunological adjuvant using or give separately disclosed any compositions in this literary composition of patient.This carcinoma of prostate can be androgen responsive type, and patient may not accept androgen-deprivation treatment.

So, another aspect of the present disclosure also relates to the method for the treatment of patient's androgen responsive type or non-sensitive type carcinoma of prostate, and the method comprises combining with immunological adjuvant to be used or gives separately patient above-mentioned any compositions.

Brief Description Of Drawings

Fig. 1 has shown the various epi-positions that in embodiment 1 HLA binding peptide mixture used, HLA binding peptide contains, and comprises 13 kinds of epi-positions in prostate related antigen molecule source.Sequence numbering: 1 to sequence numbering: 12 epi-position is HLA I class epi-position, i.e. HLA-A*201-restricted epitope.Sequence numbering: 13 to sequence numbering: 14 epi-position is HLA II class epi-position.

Fig. 2 provides the DT statistics of doing as a whole seminar.DT represents with a month number." n " refers to that statistics is included in the patient's of each apoplexy due to endogenous wind quantity." % " refers to the patient's who is divided into each apoplexy due to endogenous wind percentage ratio.When research finishes, because No. 5 patients' doubling time is born, so its doubling time do not include in geometrical mean or range computation.

Fig. 3 has shown that patient uses before HLA binding peptide mixture treatment used in embodiment 1, the DT during treatment and after treatment changes.On the occasion of representing the PSA doubling time, refer to that patient's PSA level is raising.Negative value represents the PSA half-life, refers to that patient's PSA level is declining.

Fig. 4 has shown by the treatment clinical response of immunological adjuvant type classification.Without clinical response, with "-" symbol, represent.The interim PSA that PSA accelerates to raise before declines or stablizes and represents with " +/-" symbol.PSA declines and the interim PSA rising of PSA DT before increasing represents with " /+" symbol.PSA DT increase represents with "+" symbol.

Fig. 5 has shown the peptide being received in montanide but has not accepted adjuvant or high temperature patient's PSA level.

Fig. 6 has shown the peptide that is received in montanide and Mucin1-mRNA/ protamine complex patient's PSA level.

Fig. 7 has shown the peptide that is received in montanide and high temperature patient's PSA level.

Fig. 8 has shown the peptide that is received in montanide and GM-CSF patient's PSA level.

Fig. 9 has shown the peptide that is received in montanide and imiquimod patient's PSA level.

Figure 10 has shown the variation the 84th day PSA value from reference value (representing with percentage ratio).

Figure 11 has shown the variation of PSA value (representing with percentage ratio) between seed stage.

Figure 12 has shown after inoculation has T cell to every kind of aitiogenic patient's quantity of peptide.

Figure 13 has shown CD4+T cell from the 15 and No. 26 patient PBMC specificity to PSMA459-473 epi-position and survivin 97-111 epi-position.PSMA:PSMA459-473 epi-position.Survivin: survivin 97-111 epi-position.

Figure 14 has shown the non-specific activation of reactive polypeptide=non-antigen dependency of clone Pro26_1C.PMA/ ionomycin; Survivin (II): stimulate by survivin 97-111 epi-position; PSMA (II): stimulate by PSMA459-473 epi-position.All reacting cells are that CD4 is positive.

Figure 15 has shown the non-specific activation of reactive polypeptide=non-antigen dependency of clone Pro15_10O.PMA/ ionomycin; Survivin (II): stimulate by survivin 97-111 epi-position; PSMA (II): stimulate by PSMA459-473 epi-position.All reacting cells are that CD4 is positive.

Figure 16 has described the response feature of CD4+ survivin specific T-cells clone Pro15_ " D " to the dendritic cell stimulating by total length survivin or survivin 97-111 epi-position.As shown in intracellular cytokine dyeing, the immature dendritic cell of hatching with restructuring survivin albumen or survivin 97-111 epi-position can be vaccinated patient's survivin specific T-cells identification.These results show, survivin is by protease natural decomposition in dendritic cell, and catabolic process can not destroy survivin 97-111 epi-position.In addition, these CD4+T cells are multi-functional, because their secrete cytokines IFN-γ, TNF-α and IL-2 have the CD40L (CD154) of surface expression, and carry out the threshing shown in CD107a surface expression.Shown t cell responses has antigenic specificity, because the dendritic cell ciita that T cell is not hatched by nothing to do with albumen RAP80 or HIV-001 peptide.

Figure 17 has shown the reaction of CD4+ survivin specific T-cells clone Pro26-10-C to the dendritic cell stimulating by total length survivin, survivin 97-111 epi-position or PSMA459-473 epi-position.As shown in intracellular cytokine dyeing, the immature dendritic cell of hatching with restructuring survivin albumen, survivin 97-111 epi-position and PSMA459-473 epi-position can be vaccinated patient's T cells with antigenic specificity identification.Although survivin and PSMA epi-position are all responded, the reaction that T cell stimulates survivin epi-position is stronger.These CD4+T cells are multi-functional, because their secrete cytokines IFN-γ, TNF-α and IL-2 have the CD40L (CD154) of surface expression, and carry out the threshing shown in CD107a surface expression.Shown t cell responses is antigenic specificity, because the dendritic cell ciita that T cell is not hatched by nothing to do with albumen RAP80.

Figure 18 has shown the reaction of the dendritic cell that CD4+ survivin specific T-cells clone Pro26-10-C stimulates the survivin albumen of the psma protein of processing with total length psma protein, E.C. 3.4.21.64 or E.C. 3.4.21.64 processing.

Figure 19 has shown reacting of CD4+ survivin specific T-cells Pro26-10-C pair of fixedly dendritic cell of hatching with total length survivin, survivin 97-111 epi-position, total length PSMA or PSMA459-473 epi-position of clone.As shown in intracellular cytokine dyeing, with survivin 97-111 epi-position but not the fixedly dendritic cell that total length survivin is hatched can be vaccinated patient's T cells with antigenic specificity identification.This confirmation, in order to have antigenicity, total length survivin must be by antigen presenting cell cracking.

Figure 20 has shown the reaction of CD4+PSMA specific T-cells clone Pro26-10-D to the dendritic cell stimulating by total length survivin, survivin 97-111 epi-position, total length PSMA or PSMA459-473 epi-position.As shown in intracellular cytokine dyeing, the immature dendritic cell of hatching with PSMA459-473 epi-position can be vaccinated patient's T cells with antigenic specificity identification.These CD4+T cells are multi-functional, because their secrete cytokines IFN-γ, TNF-α and IL-2 have the CD40L (CD154) of surface expression, and carry out the threshing shown in CD107a surface expression.Shown t cell responses has antigenic specificity, because T cell is not by the dendritic cell ciita of hatching with survivin or survivin 97-111 epi-position.

Figure 21 has shown that expressing the allelic several tumor cell lines of different HLA-DR identifies (as shown in patient Pro26 and Pro15) with patient's PBMC.Patient produces polyclone t cell responses after inoculating with survivin 97-111.Survivin 97-111 shows and several HLA II class allele mixes combination: DR1; (referring to the article of Wang etc.); DQ5(Wang etc. do not detect); DR11(is referring to the article of Wang etc.); Or the article table 1 of 2008 such as DRB3(and Wang etc. is contrary).In several HLA II quasi-molecules, the functional displaying of survivin 97-111 is possible (generating TNF-α).As for HLA I class (HLA-A ,-B, C), also can find three different genes sites, i.e. HLA-DQ, HLA-DP and HLA-DR for the HLA II class at the functional II quasi-molecule of cell surface expression in principle.I quasi-molecule be by heavy chain (A ,-B ,-C) and in three genes all constant beta-2-microglobulin form.Yet II quasi-molecule is comprised of two chains (α and β) of variable chains.Therefore, II quasi-molecule makes complicated gene Clustering become more complicated.In the drawings, provided the so-called serology type based on antibodies.For instance, so the not iso-allele that " DQ3 " contains HLA-DQ α and β chain, these allele conventionally occur together, react with specific antibodies.In figure, cell is as follows: and AL=E418EBV conversion B cell line (< < human immunity is learned > >, and 1996,11,51(1): 13-22); LAM=B lymphoma cell line (< < cancer genomics > >, 2002,9,19,21(42): 6549-6556); HO301=EBV conversion B cell line (< < Journal of Immunology > >, 1998,160:3363-3373); BM15=Dr11+APC cell line (< < Journal of Immunology > >, 2004,173:1876-1886); MGAR=homotype in conjunction with B-LCL (< < gene therapy > >, 2004,11,1408-1415); The autologous B cell line of LG2-EBV=(< < Cancer Immunity > >, Vol.2, p.9 (2002,7,19)); EMJ=B lymphoblastoid cell lines (European zooblast storehouse numbering: 8602103IHW numbering 9097: and < < Hum Immunol > >, 1980,12,1 (4): 363-8).

Describe in detail

The disclosure relates to compositions and method used in prostata tissue and/or the immunoreation of tumor of prostate specific antigen peptide control prostate gland cancer cell.

Whether can depend on whether there is the antigen that is considered as foreign body by host immune system by immune response stimulating.In recent years, successfully identified (1) in prostata tissue or tumor of prostate specifically expressing and (2) by the various antigens of T cell recognition.When these antigens are expressed as the complex of HLA molecule and peptide in antigen presenting cell (APC), they can stimulate T cell and induce the T cells with antigenic specificity reaction of prostate gland cancer cell, finally cause tumor cell cracking.For example, the various epi-positions of albumen survivin have been confirmed as prostate TAA.Now, the discovery of prostate specific TAA and evaluation have improved the probability of using host immune system intervention tumor growth.For immunotherapy for cancer, exploring at present the body fluid controlled in immune system and the various mechanism of cellular immunization.

The element-specific of cell immune response is to identify specifically and to destroy tumor cell.From tumor-infiltrated cell mass or peripheral blood, isolate CD8+ cytotoxic T cell (" CTL ") and show, these cells play a significant role in the innate immune defence of cancer.CD8+T cell particularly, in this reaction, play a significant role, it can identify the I quasi-molecule of 8 to 10 peptides that residue forms in the Cell sap of major histocompatibility complex (MHC) load protein or damaged ribosome product (DRIPS) source.(Schubert U, Ant ó n LC, Gibbs J, Norbury CC, Yewdell JW, Bennink JR.; Rapid degradation of a large fraction of newly synthesized proteins by proteasomes. < < nature > > (Nature, 2000; 404 (6779): 770-774).

Angtigen presentation is to be undertaken by a class of two large class major histocompatibility complex (MHC) molecule MHC I classes and MHC II apoplexy due to endogenous wind.For the mankind, MHC molecule is called as human leukocyte antigen (" HLA ") molecule.MHC molecule has two classes: MHC I quasi-molecule, on most of nucleated cell, can find this quasi-molecule, and it offers the peptide mainly producing because of endogenous, Cytoplasm or Nuclear extract matter, DRIPS and larger peptide protein cleavage.Yet the peptide that is derived from interior body structure or exogenous source is also found on the MHC I quasi-molecule of being everlasting.The non-classical mode of offering of this I quasi-molecule is called as to intersect in the literature offers.MHC II quasi-molecule, is mainly found in full-time antigen presenting cell (APC) upper, and mainly offers by APC, to be absorbed the extrinsic protein peptide of also processing subsequently in endocytosis process.For I quasi-molecule, the additive method of antigen processing is described as: it allows MHC II quasi-molecule to offer from exogenous peptide (as: autophagocytosis).The complex of peptide and MHCI quasi-molecule is identified by the corresponding TCR of CD8+ cytotoxic T lymphocyte, and the complex of peptide and MHC II quasi-molecule is identified by the corresponding TCR of the positive helper T cell of CD4.

HLA I quasi-molecule can find in each nucleated cell of human body, and its function is to CD8+ cytotoxic T cell (" CTL "), to show the fragment of cytoplasmic protein.Protein is cracking in proteasome, and the peptide of generation is transported kytoplasm and sends in endoplasmic and be combined with HLA I quasi-molecule.Then, the complex of antigen and HLA I quasi-molecule is transported to antigen and can be offered the cell surface to CTL.Angtigen presentation causes cascade reaction to CTL, finally causes directly killing the Peptide-specific CTL amplification of surface combination antigen/HLA complex cell.This process occurs in each nucleated cell, thereby can allow immune system accurately monitor existence foreign body, change or embryonated egg white matter in each cell.Although the peptide that HLA I-quasi-molecule is offered mainly, from endogenous intracellular protein molecule, there are indications, cell is drunk by giant cell or the exogenous antigen of phagocytosis endocytosis also can be offered.Therefore, can inducing antigen-specific ctl response with the polypeptide direct immunization host containing HLA I class specificity epitope.

Different from the HLA I quasi-molecule of structural expression, HLA II quasi-molecule almost only finds in full-time antigen presenting cell, comprises macrophage, dendritic cell and B cell.Full-time APC is by full-time APC endocytosis extracellular protein and digests in lysosome, at molecular migration, before plasma membrane, is combined with HLA II quasi-molecule.HLA II class binding peptide is offered to CD4+T accessory cell (" t helper cell ").T helper cell, without any direct cytotoxic activity or activate the phagocytic capacity, therefore can not directly kill the host cell of infected or functional disorder, but can to reaction infected or functional disorder cell, work by inducing or increasing other immune system compositions.For instance, CD4+T cell activation can cause the local rising of interferon-γ (IFN γ) level.

The positive helper T cell of CD4 plays a significant role in the effector function that arranges antitumor t cell responses, and therefore, it is extremely important that the TAA identification in CD4 positive T cell epi-position source may be split the drug products of the anti tumor immune response of carrying out the coffin upon burial.In mammal model, prove, even in the situation that there is no CTL, the merocrine interferon-γ of t helper cell (IFN γ) also can suppress tumor development by suppressing angiogenesis.Consult Qin, Z.and T.Blankenstein.CD4+T-cell--mediated tumor rejection involves inhibition of angiogenesis that is dependent on IFN gamma receptor expression by non-hematopoietic cells.Immunity.2000,12:677-686.In addition, the t helper cell of identification HLA II TAA that quasi-molecule is shown can be offset tumour progression (Kennedy by induction antibody response, R.C., M.H.Shearer, A.M.Watts, and R.K.Bright.CD4+T lymphocytes play a critical role in antibody production and tumor immunity against simian virus40large tumor antigen.Cancer Res.2003,63:1040-1045).

Different in conjunction with TAA from HLA-I class, only a small amount of HLA II class was had to description (www.cancerimmunity.org, www.syfpeithi.de) in conjunction with TAA so far.Because the structural expression of HLA II quasi-molecule only limits to immune system cell conventionally, thus directly from primary tumor separated II class peptide be considered to impossible.But having recently a large amount of MHC II class epi-positions is directly to determine from tumor.And, in tumor patient, be surprised to find tumor cells expression MHC II quasi-molecule (EP1642905, EP1760088; Dengjel J, Nastke MD, Gouttefangeas C, Gitsioudis G, Schoor O, Altenberend F, M ü ller M, b, Missiou A, Sauter M, Hennenlotter J, Wernet D, Stenzl A, Rammensee HG, Klingel K,

s.; Unexpected abundance of HLA class II presented peptides in primary renal cell carcinomas; Clin Cancer Res.2006; 12:4163-4170).

The peptide that triggers cell immune response must be attached on MHC molecule.This process depends on the allele of MHC molecule and the specificity polymorphism of peptide ammino acid sequence.MHC I class binding peptide length is generally 8-10 amino acid residue, is conventionally having two conserved residues (" deadman ") with the interactional sequence of the corresponding engagement groove of MHC molecule.Like this, each MHC allele have one determine which peptide can specific binding to " binding motif " (Rammensee HG, Bachmann J of engagement groove, Stevanovic S.MHC ligands and peptide motifs, Landes Bioscience, USA, 1997).

In the reaction of MHC dependent immunity, peptide not only must be combined with some MHC molecule of tumor cells expression, and they also must be able to be identified by the specific t-cell receptor of T cell (TCR).

The antigen of tumor specific cytotoxic T lymphocyte identification, that is to say, their epi-position, can be the molecule of all proteins apoplexy due to endogenous wind, as, enzyme, receptor, transcription factor etc.In addition, tumor associated antigen also can only be present in tumor cell, in mutant gene product.Another kind of important tumor associated antigen is tissure specific antigen, as, be expressed in " cancer testis " in dissimilar tumor and healthy testis tissue (CT) antigen.

In recent years, existing a large amount of TAA are successfully identified and are identified.In addition, aspect other tumor associated antigens of identification, also doing large quantity research.Some tumor associated antigen, in the art also referred to as tumor specific antigen, has tissue specificity.Example includes, but is not limited in the PSA of melanomatous tryrosinase, carcinoma of prostate and PSMA, lymphoma chromosomal chiasma (transposition) position such as bcr/abl.But; manyly obtain definite tumor associated antigen and come across in polytype tumor; some antigen; as; in fact (tumor suppressor gene is Linehan WM to cause the carcinogenic protein of transformation event and/or tumor suppressor gene; Walther MM, Zbar B is about renal carcinoma summary The genetic basis of cancer of the kidney.J Urol.2003Dec; 170 (6Pt1): mentioned in 2163-72) almost come across in all types tumor.For example; control the normal cell protein of Growth of Cells and differentiation; as p53(tumor suppressor gene), ras, c-met, myc, pRB, VHL and HER-2/neu can accumulate sudden change; the up-regulated that causes these gene outcomes; thereby carcinogenic (McCartey et al.Cancer Research; 1998,15:582601-5; Disis et al.Ciba Found.Symp.1994,187:198-211).These mutains are the immunoreactive target of tumour-specific of kinds cancer also.

Effective or invalidly offered to determine type and the degree of institute's induction of immunity reaction from immunity to toleration.In order to stimulate dormancy, unsensitized CTL, CTL must receive two signals of antigen presenting cell in antigenic specificity mode.One be by with the interactional T cells with antigenic specificity receptor of HLA/ peptide complexes (TCR).Another is by costimulating factor (B7 molecule, ICAM-1 and other adhesion molecules) or cytokine (as IL-2).When T lymphocyte is only received one of them signal, T cell anergy.The normally weak APC of tumor cell, because they do not have costimulatory molecules, conventionally only manifests low HLA I class and expresses.In addition, the common expression inhibiting of malignant cell is for them immunoreactive cytokine or surface molecular.Similarly, TAA offers to the mode of T cell significant to inducing tumor-specific immunoreation.

For be identified as the protein of tumour-specific or related antigen by CTL, in treatment, use, must meet specific prerequisite.This antigen should be mainly by tumor cells expression, and normal health is organized and do not expressed or express negligible amounts.More suitable situation is that this corresponding antigens not only comes across in a kind of tumor, and concentration (being the corresponding peptides copy number of each cell) height.Tumor specific antigen and tumor associated antigen normally come from because the functions such as cell cycle regulating or apoptosis are converted into the albumen of directly getting involved in tumor cell at normal cell.In addition, these directly cause the downstream targets of the albumen of conversion also may to be raised, therefore indirect and Tumor-assaciated.These indirect tumor associated antigens may be also the targets of prophylactic immunization method.This peptide from tumor associated antigen (" immunogenic peptide ") it is essential, in both of these case, all have the epi-position of antigen aminoacid sequence, so can cause t cell responses in external or body.

Substantially, any peptide that can be combined with MHC molecule all may serve as a t cell epitope.In the outer or body of inductor, the prerequisite of t cell responses is to have the immunologic tolerance that has the T cell of corresponding TCR and there is no this defined epitope.Helper T cell plays an important role in the CTL of layout antineoplastic immune effector function.Trigger T _h1the helper T cell epitope of cell effect is supported the effector function of the positive killer T cell of CD8, comprising the cytotoxicity function (such tumor cell surface shows tumor-associated peptides/MHC complex) that directly acts on tumor cell.Like this, separately form or can be used as with the Tumor-assaciated t helper cell peptide epitopes that other tumor-associated peptides form compositionss the active pharmaceutical ingredient that stimulates anti tumor immune response vaccine combination.

For most of tumors, to only have minority TAA be known or only defined the TAA of a specific HLA type.In contrast, a large amount of prostate specific antigen of CTL identification and carcinoma of prostate related antigen and peptide are successfully identified.When these TAA are expressed as the complex of HLA molecule and peptide in antigen presenting cell (APC), they can stimulate T cell inducing antigen-specific CTL.

In research in the most of experience of immunization therapy from malignant melanoma, show can cause tumor cell selection with one or two TAA immunity, thereby cause PD in DC continued treatment process.Because the common and collaborative antitumous effect that produces of two kinds of reactions that CD8 and CD4 rely on, so CD8+CTL(MHC I quasi-molecule) or the positive CTL(MHC II of CD4 quasi-molecule) tumor associated antigen identified identification and identify in development tumor vaccine very important.

But, excite a kind of CTL to be conventionally not enough to remove all tumor cells.Tumor is very easy to mutagenesis, thereby thereby the identification that can hide CTL by the protein profiling that changes them fast CTL is attacked and is made a response.In order to strike back tumor escape mechanism, in vaccination, use various specific peptides.Like this, several ctl clone can initiate to walk abreast attack on a large scale to tumor simultaneously.This may reduce the chance that the reaction of tumor escape from immune is hit.This hypothesis is confirmed recently in a clinical research for the treatment of melanoma patients in late period.Only there is a few exceptions situation, have at least the patient of three kinds of different t cell responses to occur objective clinical response or stable disease (Banchereau etc., 2001) and survival rate raise, t cell responses is less than the Most patients of three kinds and is diagnosed with PD at the same time.

When using the vaccine therapy that contains 13 kinds of different peptides, renal cell carcinoma patients has similar effect (H.Singh-Jasuja, S.Walter, T.Weinschenk, A.Mayer; P.Y.Dietrich, M.Staehler, A.Stenzl; S.Stevanovic, H.Rammensee, J.Frisch; Correlation of T-cell response; clinical activity and regulatory T-cell levels in renal cell carcinoma patients treated with IMA901, a novel multi-peptide vaccine; ASCO meeting 2007Poster#3017; M.Staehler, A.Stenzl, P.Y.Dietrich, T.Eisen, A.Haferkamp, J.Beck, A.Mayer, S.Walter, H.Singh, J.Frisch, C.G.Stief; An open label study to evaluate the safety and immunogenicity of the peptide based cancer vaccine IMA901, ASCO meeting 2007; Poster#3017).

Therefore, develop a kind of main task of tumor vaccine and not only will identify and identify that novel tumor related antigen and immunogen T thereof assist epi-position, but also will combine different epitopes to increase every patient to the aitiogenic probability of more than one epi-positions.

Therefore, provide the compositions that comprises at least one HLA binding peptide, HLA binding peptide comprises, mainly by or by the epi-position in prostate related antigen molecule source, formed.

Term used in literary composition " HLA binding peptide " refers to any polypeptide that can be combined with HLA I class or HLA II class human leukocyte antigen molecule.

In literary composition, term used " epi-position " refers to the aminoacid sequence that can be enough to allow the molecule that contains epi-position be combined with HLA I class or HLA II class human leukocyte antigen molecule.The method of identification epi-position is method known in technique.These methods include but not limited to: (a) utilize the T cell of single patient to carry out gene expression analysis; referring to van der Bruggen et al.; A gene encoding an antigen recognized by cytolytic T lymphocytes on a human melanoma, Science1991; 254 (5038): 1643 – 1647; (b) mass spectrum of tumor-associated peptides order-checking, Cox et al., Identification of a peptide recognized by five melanoma-specific human cytotoxic T cell lines, Science1994; 264 (5159): 716 – 719; (c) " oppositely immunology "; according to known TAA prediction epi-position for allele-specific peptide motif; referring to Stevanovic, Identification of tumor-associated T-cell epitopes for vaccine development, Nat.Rev.Cancer 2002; 2 (7): 514 – 520; Celis et al.; Induction of anti-tumor cytotoxic T lymphocytes in normal humans using primary cultures and synthetic peptide epitopes, Proc.Nat ' l Acad.Sci.USA1994; 91 (6): 2105 – 2109.

Term used in literary composition " prostate related antigen molecule " refers to and contains (a) prostata tissue or (b) any epi-position of prostate gland cancer cell differential expression molecule.Typical prostate related antigen molecule comprises: prostate specific antigen, prostate stem cell antigen, prostate specific membrane antigen, survivin, prostate specific albumen and transient receptor potential-p8.From the typical epi-position of these prostate related antigen molecules as shown in Figure 1.And, can use any epi-position in any prostate related antigen molecule source.

If there is no other explanations, in literary composition, disclosed HLA binding peptide can be by peptide chain different loci, may be that selectivity site is replaced one or more residues and modified.This replacement may be conservative, and for example, one of them aminoacid is had another aminoacid of similar structures and feature to be replaced, such as one of them hydrophobic amino acid is replaced by another hydrophobic amino acid.More conservative replacement is the replacement having between the aminoacid of same or similar size and chemical property, and for example, leucine is replaced by isoleucine.In the research of natural homologous protein family sequence variations, some amino acid substitution is more often tolerated than other amino acid substitution, and this often shows with the similarity on size, electric charge, polarity and hydrophobicity between original aminoacid and its alternative relevant.

In this article, conservative is replaced to be defined as in the inside of one of following five kinds of groups and is exchanged: group 1-little aliphatic, the nonpolar or residue (Ala, Ser, Thr, Pro, Gly) of tool polarity slightly; Group 2-polarity, electronegative residue and amide (Asp, Asn, Glu, Gln) thereof; Group 3-polarity, positively charged residue (His, Arg, Lys); Group 4-large aliphatic non-polar residue (Met, Leu, Ile, Val, Cys) and group 5-large aromatic residues (Phe, Tyr, Trp).

More not conservative replace may relate to that an aminoacid is had similar feature by another but in size different aminoacid replace, as: alanine is replaced by isoleucine residue.Highly the replacement of conservative may not relate to the aminoacid that an acidic amino acid has polarity by another or even have an alkaline nature and replaces.Yet this " radical " replaced and can not be thought invalid and do not consider, because chemical action is not exclusively predictable, radical property is replaced may bring the accidental effect that cannot predict in its simple principles of chemistry.

Certainly, this replacement may relate to other structure outside common L-aminoacid.Therefore, may replace L-aminoacid common in disclosed antigenic peptides with D-aminoacid, this should comprise within the scope of the present disclosure.In addition, having the aminoacid (that is, the R group except 20 common amino acids of native protein) of non-standard R group also can be for replacing it object, to produce according to immunogen of the present disclosure and immunogenic polypeptide.

If find to cause the antigen active of peptide to be substantially equal to or to be greater than following definition value in more than one locational replacement, the combination of these replacements tested, to determine whether the replacement of combination produces the antigenic stack of peptide or cooperative effect.The position of simultaneously being replaced in peptide at most can not be over 4.

On the other hand, propose the compositions that comprises at least two kinds of HLA binding peptides, wherein in HLA binding peptide, had a kind of HLA of being I class peptide at least, had a kind of HLA of being II class peptide at least.

Term used in literary composition " HLA I class peptide " refers to any polypeptide that contains the epi-position of can or estimating to combine with HLA I class human leukocyte antigen molecule.For example, " HLA I class peptide " comprises the HLA-A2-restriction peptide of being combined with the specific allele of HLA-A2 serotype, includes but not limited to the HLA-A2 serotype molecule of HLA-A*0201, * 0202, * 0203, * 0206 or * 0207 α chain.Typically " HLA I class peptide " is sequence numbering: 1,2,3,4,5,6,7,8,9,10,11, and each is HLA-A*201 restriction peptide.

The disclosure has further proposed such peptide: they derive from tumorigenesis related antigen and have the ability and the abundant combination of MHC (HLA) II quasi-molecule, trigger human leukocyte immunoreation, particularly lymphocyte, T lymphocyte, CD4 positive t lymphocytes, the immunoreactive CD4 positive t lymphocytes of mediation Th1 type.Term used in literary composition " HLA II class peptide " refers to and contains any polypeptide of can or estimating to be combined with HLA II class human leukocyte antigen molecule epi-position.Typically " HLA II class peptide " listed in Fig. 1 by sequence numbering: sequence numbering: 13 and sequence numbering: 14.

Other peptides of invention are:

Well-known HLA II class peptide is by holding extension area (being regarded as with peptide and T cell interaction irrelevant) to form containing specific HLA specific amino acid motif " core sequence " and the optional N-of core sequence function and/or the C-of not disturbing.For example, the length of N-and/or C-end extension area can be respectively between 1 to 10 aminoacid.These peptides can be directly used in loading HLA II quasi-molecule or its sequence can be cloned into hereinafter described carrier.Because these peptides can form the end product of the larger peptide course of processing in cell, therefore, also available long peptide.In literary composition, disclosed HLA II class peptide can be arbitrary size, includes but not limited to that molecular weight is less than 100,000 dalton, 50,000 dalton, 10,000 dalton, 5,000 dalton, 2,500 dalton or from approximately 1,000 dalton to 2,000 dalton.About the quantity of amino acid residue, for instance, the peptide of not getting rid of in the disclosure may have less than 1000,500 or 100 residues.

Therefore, the compositions of peptide and variant thereof is disclosed, and wherein the total length of peptide or variant is 8 to 100,8 to 60,8 to 30 and 8 to 17 aminoacid, or wherein peptide or variant have 8,9,10,11,12,13,14,15 or 16 aminoacid.On the other hand, peptide contains the core sequence of selecting from following one group of sequence numbering: sequence numbering: 13 and 14, there is 1 to 10 aminoacid extension area at C-end and/or N-end, wherein flanking amino acid adds up to 1 to 12, 1 to 10, 1 to 8, 1 to 6, 2 to 12, 2 to 10, 2 to 8, 2 to 6, 3 to 12, 3 to 10, 3 to 8, 3 to 6, 4 to 12, 4 to 10, 4 to 8, 4 to 6, 5 to 12, 5 to 10, 5 to 8, 5 to 6, 6 to 12, 6 to 10, 6 to 8, 7 to 12, 7 to 10, 7 to 8, 8 to 12, 8 to 10, 9 to 12, 9 to 10 or 10 to 12, flanking amino acid can any ratio be distributed in C-end and N-holds (as all flanking amino acids can add at one end, or aminoacid is average or be added in two ends with any other ratio), but this peptide still can be attached on HLA molecule in the same mode of described peptide.

MHC I class epi-position (normal length is 8 to 10 aminoacid) also may be processed and produce by peptide from the peptide compared with long or the albumen that comprises actual epi-position.Similar with MHC II class epi-position, may select actual epi-position N-and C-end to extend the flank residue in precursor peptide upstream and/or downstream, they can not significantly affect peptide and offer can not hide essential protease cracking site and generate the actual epi-position that extends the mediation of peptide machining to CTL like this.

So on the other hand, peptide contains the core sequence that following sequence numbering forms: sequence numbering: 1 to 11 and sequence numbering: 15 to 37, at C-end and/or N-end extension area, there is 1 to 10 flanking amino acid.On the other hand, these flanking amino acids add up to 1 to 12, 1 to 10, 1 to 8, 1 to 6, 2 to 12, 2 to 10, 2 to 8, 2 to 6, 3 to 12, 3 to 10, 3 to 8, 3 to 6, 4 to 12, 4 to 10, 4 to 8, 4 to 6, 5 to 12, 5 to 10, 5 to 8, 5 to 6, 6 to 12, 6 to 10, 6 to 8, 7 to 12, 7 to 10, 7 to 8, 8 to 12, 8 to 10, 9 to 12, 9 to 10 or 10 to 12, wherein flanking amino acid can any ratio be distributed in C-end and N-holds (as all flanking amino acids can add at one end, or aminoacid is average or be added in two ends with any other ratio), but the mode that this peptide still can be same with the peptide of described any sequence numbering is attached on HLA molecule: sequence numbering: 1 to 12 and sequence numbering: 15 to 40.

On the other hand, it is 8 to 100,8 to 60,8 to 30 and 8 to 17 aminoacid that the disclosure has proposed total length, or has peptide and the variant of 8,9,10,11,12,13,14,15,16 or 17 amino acid whose MHC I class epi-positions.

Certainly, disclosed peptide or variant can be attached on people MHC I or II quasi-molecule.Peptide or variant and MHC complex associativity can be tested with the known method in this area; for example; the allelic method of the different MHC II classes of detection described in method described in the following example of the present invention or document (for example; Vogt AB, Kropshofer H, Kalbacher H; Kalbus M; Rammensee HG, Coligan JE, Martin R; Ligand motifs of HLA-DRB5*0101and DRB1*1501molecules delineated from self-peptides; J Immunol.1994; 153 (4): 1665-1673; Malcherek G, Gnau V, Stevanovic S, Rammensee HG, Jung G, Melms A; Analysis of allele-specific contact sites of natural HLA-DR17ligands; J Immunol.1994; 153 (3): 1141-1149; Manici S, Sturniolo T, Imro MA, Hammer J, Sinigaglia F, Noppen C, Spagnoli G, Mazzi B, Bellone M, Dellabona P, Protti MP; Melanoma cells present a MAGE-3epitope to CD4 (+) cytotoxic T cells in association with histocompatibility leukocyte antigen DR11; J Exp Med.1999; 189 (5): 871-876; Hammer J, Gallazzi F, Bono E, Karr RW, Guenot J, Valsasnini P, Nagy ZA, Sinigaglia F; Peptide binding specificity of HLA-DR4molecules:correlation with rheumatoid arthritis association; J Exp Med.1995181 (5): 1847-1855; Tompkins SM, Rota PA, Moore JC, Jensen PE; A europium fluoro-immunoassay for measuring binding of antigen to class II MHC glycoproteins; J Immunol Methods.1993; 163 (2): 209-216; Boyton RJ, Lohmann T, Londei M, Kalbacher H, Halder T, Frater AJ, Douek DC, Leslie DG, Flavell RA, Altmann DM; Glutamic acid decarboxylase T lymphocyte responses associated with susceptibility or resistance to type I diabetes:analysis in disease discordant human twins, non-obese diabetic mice and HLA-DQ transgenic mice; Int Immunol.1998 (12): 1765-1776).

Other N of aminoacid and/or C end extended spot differ and form surely the peptide as MHC molecule epi-position, but have important function to effectively the peptide in the disclosure being introduced to cell.In one embodiment, peptide of the present disclosure is fusion rotein, it is containing the invariant chain (p33 that is correlated with just like HLA-DR antigen, be designated hereinafter simply as " Ii ") 80 aminoacid of N-end, derive from NCBI, gene bank number of registration X00497 (Strubin, M., Mach, B.and Long, E.O.The complete sequence of the mRNA for the HLA-DR-associated invariant chain reveals a polypeptide with an unusual transmembrane polarity.EMBO is (4) J.3,869-872 (1984)).

On the other hand, the total length of peptide is 8 to 100,8 to 60,8 to 30 and 8 to 17 aminoacid, or has 9,10,11,12,13,14,15 or 16 aminoacid.

In addition, this peptide or variant can further be modified to improve stability and/or be combined with MHC molecule, thereby cause stronger immunoreation.Such optimization method of peptide sequence is known in this area, comprises, for example, the introducing of trans peptide bond and non-peptide bond.

Therefore, according on the other hand, the disclosure has proposed a kind of compositions, and wherein, at least one peptide or variant contain non-peptide bond.

In trans peptide bond aminoacid, peptide (CO-NH-) does not connect its residue, but its peptide bond is reverse.This converse simulating peptide (retro-inverso peptidomimetics) can be prepared by methods known in the art, such as: Meziere etc. (1997) are at (< < Journal of Immunology > > 159, method 3230-3237), is incorporated to herein by reference.This method relates to preparation and comprises the pseudo-peptide that skeleton (and being not side chain) changes.Meziere etc. (1997) prove that these pseudo-peptides are useful to MHC and t helper cell reaction.The converse peptide that substitutes CO-NH peptide bond with NH-CO key has improved resistant to hydrolysis performance widely.

Non-peptide bond is-CH ₂-NH ,-CH ₂s-,-CH ₂cH ₂-,-CH=CH-,-COCH ₂-,-CH (OH) CH ₂-and-CH ₂sO-etc.The U.S. 4,897, No. 445 patent has proposed non-peptide bond (CH in polypeptide chain ₂-NH) non-solid-phase synthesis, the method relate to by the synthetic polypeptide of standardization program and by amido aldehyde and a seed amino acid at NaCNBH ₃under the condition existing, interact and synthetic non-peptide bond.

Peptide containing above-mentioned disclosure sequence can synthesize with other chemical groups of its amino and/or carboxyl terminal, thereby improves stability, bioavailability and/or the affinity etc. of peptide.For example, the hydrophobic group such as benzyloxycarbonyl group, dansyl base or tertiary butyloxycarbonyl group can add the amino terminal of peptide.Equally, acetyl group or 9-fluorenylmethyloxycarbonyl may be positioned at the amino terminal of peptide.In addition, for example, hydrophobic group, tertiary butyloxycarbonyl group or amine groups all may be added into the carboxyl terminal of peptide.

In addition, all peptides in the disclosure all may change through synthesizing its steric configuration.For example, may use the d-isomer of one or more amino acid residues of these peptides, rather than common levo form.Further, the alpha-non-natural amino acid residue that in the disclosure, at least one amino acid residue of peptide can be known is replaced.Suchlike change may contribute to increase stability, bioavailability and/or the combination of disclosure peptide.

Equally, the peptide in the disclosure or variant can carry out chemical modification by special amino acid whose reaction before or after synthetic peptide.The embodiment of this type of modification is known in the art, for example, < < Chemical Reagents for Protein Modification > > (the 3rd ed.CRC Press showing at R.Lundblad, 2005) in, there is general introduction, in the mode of list of references, be incorporated to herein.Amino acid whose chemical modification method includes, but is not limited to modify by the following method: acyl group, amidination, lysine pyrrole tremble base, standard reductive alkylation, with 2; 4; 6-trinitro-benzene-sulfonic acid (TNBS) is carried out trinitrophenyl, the carboxyl group of cysteine is carried out amidatioon and its sulfydryl is cysteic acid, forms variable derivant, forms mix disulfide, react with maleimide with other sulfhydryl compound through performic oxidation amino, with iodoacetic acid or iodoacetamide carboxy methylation, under alkaline pH value with cyanate carbamoylation.In this respect, technical staff with reference to < < Current Protocols In Protein Science > > (Coligan etc. write. (John Wiley & Sons NY1995-2000)) in described in the 15th chapter in the relevant extensive method of chemical modification.

Human cytokines and often can extend circulating half-life containing the successful modification of the peptide of Polyethylene Glycol, and use glutaraldehyde, polyethyleneglycol diacrylate and formaldehyde are cross-linked for preparing hydrogel albumen.Allergen chemical modification for immunization therapy often realizes by the carbamyl of potassium cyanate.

In general, peptide and variant (at least containing the peptide between amino acid residue, connecting) can be used the people such as Lu (1981) (< < J.Org.Chem. > >, 46,3433) and the disclosed methods such as the synthetic Fmoc-polyamide pattern of solid-phase peptide of the list of references listed of this article synthesize.

Purification can be realized by any or combined method of following technology, as: recrystallize method, size exclusion chromatograph method, ion exchange chromatography, hydrophobic interaction chromatography and (conventionally) reversed phase high-performance liquid chromatography (as used acetonitrile/water gradient separations).

Can use thin layer chromatography, electrophoresis particularly amino acid analysis, fast atom bombardment (FAB) mass spectral analysis and MALDI and the ESI-Q-TOF mass spectral analysis after capillary electrophoresis, Solid-Phase Extraction (CSPE), reversed phase high-performance liquid chromatography, acidolysis carry out peptide analysis.

The disclosure has proposed the nucleic acid (as polynucleotide) of one of the open peptide of coding or variant on the other hand.Polynucleotide may or be strand and/or two strands for DNA, cDNA, PNA, CAN, RNA etc., or the primary or stable form of polynucleotide, as: the polynucleotide with D2EHDTPA skeleton, or its compositions, and as long as its encoded peptide just may comprise also and may not comprise intron.Certainly, the peptide that polynucleotide can only be encoded and be added natural peptide bond and contain natural amino acid residue.Another aspect of the present disclosure has also proposed to express the expression vector of disclosure polypeptide.The expression vector of dissimilar cell is known in the art and just can selectes without undo experimentation.

In general, DNA can be suitable direction and correct expression reading frame insert in a kind of expression vector (as plasmid).If necessary, corresponding the transcribing with translating that this DNA may identify with required host regulates control nucleotide sequence to be connected, although general in expression vector, exists this type of to control function.Then, the standard technique of utilizing affiliated field to know imports carrier in host.

In one embodiment, compositions comprises at least two kinds of peptides that are mainly comprised of following sequence numbering aminoacid sequence: sequence numbering: 1 to 11 and sequence numbering: 13 to 40.

In another embodiment, compositions comprises at least two kinds of peptides that are mainly comprised of following sequence numbering aminoacid sequence: sequence numbering: 1,2,5,7,9 to 11 and 13 to 40.

In another embodiment, compositions comprises at least 4 kinds of peptides that are mainly comprised of following sequence numbering aminoacid sequence: sequence numbering: 1,2,5,7,9 to 11 and 13 to 40.

In another embodiment, compositions comprises at least 10 kinds of peptides that are mainly comprised of following sequence numbering aminoacid sequence: sequence numbering: 1,2,5,7,9 to 11 and 13 to 40.

In another embodiment, compositions comprises at least 2, 4 or 10 kind of peptide, this peptide is comprised of the aminoacid sequence of following sequence numbering substantially: sequence numbering: 1, 2, 5, 7, 9 to 11, 13 and 14, at least one peptide is to select from following one group of sequence numbering: from survivin HLA-A1 sequence numbering: 38 BIR-002b FTELTLGEF, from survivin HLA-A2 sequence numbering: 39 BIR-002c LMLGEFLKL, from survivin HLA-B35 sequence numbering: 40 BIR-002d EPDLAQCFY, from survivin HLA-DR BIR-002a sequence numbering: 41 TLGEFLKLDRERAKD and from the BIR-004 sequence numbering of survivin HLA-DR and HLA-A*02: 42 ELTLGEFLKLDRERAKN.

In another embodiment, compositions comprises at least 2, 4 or 10 kind of peptide, this peptide is comprised of the aminoacid sequence of following sequence numbering substantially: sequence numbering: 1, 2, 5, 7, 9 to 11, 13 and 14, at least one peptide is to select from following one group of sequence numbering: from survivin HLA-A1 sequence numbering: 38 BIR-002b FTELTLGEF, from survivin HLA-A2 sequence numbering: 39 BIR-002c LMLGEFLKL, from survivin HLA-B35 sequence numbering: 40 BIR-002d EPDLAQCFY, from survivin HLA-DR BIR-002a sequence numbering: 41 TLGEFLKLDRERAKD and from survivin HLA-DR and HLA-A*02BIR-004 sequence numbering: 42 ELTLGEFLKLDRERAKN, wherein, extra peptide is to select according to experimenter HLA group to be treated.

In one embodiment, compositions comprises at least two kinds of peptides that are comprised of following sequence numbering aminoacid sequence: sequence numbering: 1 to 11 and sequence numbering: 13 to 42.

In another embodiment, compositions comprises at least two kinds of peptides that are comprised of following sequence numbering aminoacid sequence: sequence numbering: 1,2,5,7,9 to 11 and 13 to 42.

In another embodiment, compositions comprises at least 4 kinds of peptides that are comprised of following sequence numbering aminoacid sequence: sequence numbering: 1,2,5,7,9 to 11 and 13 to 42.

In another embodiment, compositions comprises at least 10 kinds of peptides that are comprised of following sequence numbering aminoacid sequence: sequence numbering: 1,2,5,7,9 to 11 and 13 to 42.

In another embodiment, compositions comprises at least 2, 4 or 10 kind of peptide, this peptide is comprised of the aminoacid sequence of following sequence numbering: sequence numbering: 1, 2, 5, 7, 9 to 11, 13 and 14, at least one peptide is to select from following one group of sequence numbering: from the BIR-002b FTELTLGEF of survivin HLA-A1 sequence numbering 38, BIR-002c LMLGEFLKL from survivin HLA-A2 sequence numbering 39, BIR-002d EPDLAQCFY from survivin HLA-B35 sequence numbering 40, from the TLGEFLKLDRERAKD of survivin HLA-DR BIR-002a sequence numbering 41 and from the ELTLGEFLKLDRERAKN of survivin HLA-DR and HLA-A*02BIR-004 sequence numbering 42.

In another embodiment, compositions comprises at least 2, 4 or 10 kind of peptide, this peptide is comprised of the aminoacid sequence of following sequence numbering: sequence numbering: 1, 2, 5, 7, 9 to 11, 13 and 14, at least one peptide is to select from following one group of sequence numbering: from survivin HLA-A1 sequence numbering: 38 BIR-002b FTELTLGEF, from survivin HLA-A2 sequence numbering: 39 BIR-002c LMLGEFLKL, from survivin HLA-B35 sequence numbering: 40 BIR-002d EPDLAQCFY, from survivin HLA-DR BIR-002a sequence numbering: 41 TLGEFLKLDRERAKD and from survivin HLA-DR and HLA-A*02BIR-004 sequence numbering: 42 ELTLGEFLKLDRERAKN, wherein, extra peptide is according to there being the experimenter of needs HLA group to select.

In one embodiment, compositions comprises at least two kinds of peptides that are mainly comprised of following sequence numbering aminoacid sequence: sequence numbering: 1 to 11,13 to 14 and 15 to 31.

In another embodiment, compositions comprises at least two kinds of peptides that are mainly comprised of following sequence numbering aminoacid sequence: sequence numbering: 1,2,5,7,9 to 11,13,14,23,24,25,20,31 and 32.

In another embodiment, compositions comprises at least 4 kinds of peptides that are mainly comprised of following sequence numbering aminoacid sequence: sequence numbering: 1,2,5,7,9 to 11,13,14,23,24,25,20,31 and 32.

In another embodiment, compositions comprises at least 10 kinds of peptides that are substantially comprised of the aminoacid sequence that is selected from following one group of sequence numbering: sequence numbering: 1,2,5,7,9 to 11,13,14,23,24,25,20,31 and 32.

In another embodiment, compositions comprises at least 2, 4 or 10 kind of peptide, this peptide is comprised of the aminoacid sequence that is selected from following one group of sequence numbering substantially: sequence numbering: 1, 2, 5, 7, 9 to 11, 13, 14, 23, 24, 25, 20, 31 and 32, have at least a kind of peptide to be selected from following one group of sequence numbering: from survivin HLA-A1 sequence numbering: 38 BIR-002bFTELTLGEF, from survivin HLA-A2 sequence numbering: 39 BIR-002c LMLGEFLKL, from survivin HLA-B35 sequence numbering: 40 BIR-002d EPDLAQCFY, from survivin HLA-DR BIR-002a sequence numbering: 41 TLGEFLKLDRERAKD and from survivin HLA-DR and HLA-A*02BIR-004 sequence numbering: 42 ELTLGEFLKLDRERAKN.

In another embodiment, compositions comprises at least 2, 4 or 10 kind of peptide, this peptide is comprised of the aminoacid sequence of following sequence numbering substantially: sequence numbering: 1, 2, 5, 7, 9 to 11, 13, 14, 23, 24, 25, 20, 31 and 32, have at least a kind of peptide to be selected from following one group of sequence numbering: from survivin HLA-A1 sequence numbering: 38 BIR-002b FTELTLGEF, from survivin HLA-A2 sequence numbering: 39 BIR-002c LMLGEFLKL, from survivin HLA-B35 sequence numbering: 40 BIR-002d EPDLAQCFY, from survivin HLA-DR BIR-002a sequence numbering: 41 TLGEFLKLDRERAKD and from survivin HLA-DR and HLA-A*02BIR-004 sequence numbering: 42 ELTLGEFLKLDRERAKN, wherein, extra peptide is to select according to experimenter HLA group to be treated.

In an embodiment of HLA-A*02, compositions comprises at least one peptide being comprised of following sequence numbering aminoacid sequence: sequence numbering: 23 and 24, and have at least a kind of peptide to be formed by following sequence numbering aminoacid sequence: sequence numbering: 1 to 11 and sequence numbering: 13 to 14.

In an embodiment of HLA-A*02, compositions comprises at least one peptide being comprised of following sequence numbering aminoacid sequence: sequence numbering: 23,24,1,2,5,7,9 to 11,13 and 14.

In another embodiment of HLA-A*02, compositions comprises 10 kinds of peptides that are comprised of following sequence numbering aminoacid sequence: sequence numbering: 23,24,1,2,5,7,9 to 11,13,14.

In another embodiment of HLA-A*02, compositions comprises at least 2, 4 or 10 kind of peptide, this peptide is comprised of following sequence numbering aminoacid sequence: sequence numbering: 23, 24, 1, 2, 5, 7, 9 to 11, 13, 14, at least one peptide is to select from following one group of sequence numbering: from survivin HLA-A1 sequence numbering: 38 BIR-002b FTELTLGEF, from survivin HLA-A2 sequence numbering: 39 BIR-002c LMLGEFLKL, from survivin HLA-B35 sequence numbering: 40 BIR-002d EPDLAQCFY, from survivin HLA-DR sequence numbering: 41 TLGEFLKLDRERAKD and from survivin HLA-DR and HLA-A*02BIR-004 sequence numbering: 42 ELTLGEFLKLDRERAKN.

In another embodiment of HLA-A*02, compositions comprises at least 2, 4 or 10 kind of peptide, this peptide is comprised of following sequence numbering aminoacid sequence: sequence numbering 23, 24, 1, 2, 5, 7, 9 to 11, 13, 14, at least one peptide is to select from following one group of sequence numbering: from survivin HLA-A1 sequence numbering: 38 BIR-002b FTELTLGEF, from survivin HLA-A2 sequence numbering: 39 BIR-002c LMLGEFLKL, from survivin HLA-B35 sequence numbering: 40 BIR-002d EPDLAQCFY, from survivin HLA-DR BIR-002a sequence numbering: 41 TLGEFLKLDRERAKD and from survivin HLA-DR and HLA-A*02BIR-004 sequence numbering: 42 ELTLGEFLKLDRERAKN, wherein, extra peptide is according to there being the experimenter of needs HLA group to select.

In an embodiment of HLA-A*24 and HLA-A*02 combination, compositions comprises at least two kinds of peptides that are mainly comprised of following sequence numbering aminoacid sequence: sequence numbering: 20,25,31,32,1 to 11,13 and 14.

In another embodiment of HLA-A*24 and HLA-A*02 combination, compositions comprises at least two kinds of different peptides that are mainly comprised of following sequence numbering aminoacid sequence: sequence numbering: 20,25,31,32 and one groups of sequence numberings: 1,2,5,7,9 to 11,13,14,23,24,25,20,31 and 32.

In another embodiment, compositions comprises at least 2, 4 or 10 kind of peptide, this peptide is comprised of the aminoacid sequence that is selected from following one group of sequence numbering substantially: sequence numbering: 1, 2, 5, 7, to 11, 13, 14, 23, 24, 25, 20, 31 and 32, having a kind of peptide at least is to be selected from following one group of sequence numbering: from survivin HLA-A1 sequence numbering: 38 BIR-002b FTELTLGEF, from survivin HLA-A2 sequence numbering: 39 BIR-002c LMLGEFLKL, from survivin HLA-B35 sequence numbering: 40 BIR-002d EPDLAQCFY, from survivin HLA-DR sequence numbering: 41 TLGEFLKLDRERAKD and from the ELTLGEFLKLDRERAKN of survivin HLA-DR and HLA-A*02BIR-004 sequence numbering 42.

In another embodiment, compositions comprises at least 2, 4 or 10 kind of peptide, this peptide is comprised of following sequence numbering aminoacid sequence substantially: sequence numbering 1, 2, 5, 7, 9 to 11, 13, 14, 23, 24, 25, 20, 31 and 32, have at least a kind of peptide to be selected from following one group of sequence numbering: from survivin HLA-A1 sequence numbering: 38 BIR-002b FTELTLGEF, from survivin HLA-A2 sequence numbering: 39 BIR-002c LMLGEFLKL, from survivin HLA-B35 sequence numbering: 40 BIR-002d EPDLAQCFY, from survivin HLA-DR BIR-002a sequence numbering: 41 TLGEFLKLDRERAKD and from survivin HLA-DR and HLA-A*02BIR-004 sequence numbering: 42 ELTLGEFLKLDRERAKN, wherein, extra peptide is to select according to experimenter HLA group to be treated.

In one embodiment, compositions comprises at least two kinds of peptides that are comprised of following sequence numbering aminoacid sequence: sequence numbering: 20, 23 to 25, 31, 32, 1 to 11, 13 to 14, have a kind of extra peptide of following sequence numbering at least: sequence numbering: 15 to 19, sequence numbering: 21, 22, 26 to 30 and 33, wherein, extra peptide is to select according to experimenter HLA group to be treated, has at least a kind of peptide to be selected from following one group of sequence numbering (optional): from survivin HLA-A1 sequence numbering: 38 BIR-002b FTELTLGEF, from survivin HLA-A2 sequence numbering: 39 BIR-002c LMLGEFLKL, from survivin HLA-B35 sequence numbering: 40 BIR-002d EPDLAQCFY, from survivin HLA-DR BIR-002a sequence numbering: 41 TLGEFLKLDRERAKD and from survivin HLA-DR and HLA-A*02BIR-004 sequence numbering: 42 ELTLGEFLKLDRERAKN, wherein, extra peptide is selected according to experimenter HLA group to be treated.

In another embodiment, compositions comprises at least two kinds of peptides that are comprised of following sequence numbering aminoacid sequence: sequence numbering: 20, 23 to 25, 31, 32, 1, 2, 5, 7, 9 to 11, 13, 14, have a kind of extra peptide of following sequence numbering at least: sequence numbering: 15 to 19, sequence numbering: 21, 22, 26 to 30 and 33, wherein, extra peptide is according to there being the experimenter of needs HLA group to select, and has at least a kind of peptide to be selected from following one group of sequence numbering (optional): from survivin HLA-A1 sequence numbering: 38 BIR-002b FTELTLGEF, from survivin HLA-A2 sequence numbering: 39 BIR-002c LMLGEFLKL, from survivin HLA-B35 sequence numbering: 40 BIR-002d EPDLAQCFY, from survivin HLA-DR BIR-002a sequence numbering: 41 TLGEFLKLDRERAKD and from survivin HLA-DR and HLA-A*02BIR-004 sequence numbering: 42 ELTLGEFLKLDRERAKN, wherein, extra peptide is to select according to experimenter HLA group to be treated.

In another embodiment, compositions comprises at least 4 kinds of peptides that are comprised of following sequence numbering aminoacid sequence: sequence numbering: 20, 23 to 25, 31, 32, 1, 2, 5, 7, 9 to 11, 13, 14, have a kind of extra peptide of following sequence numbering at least: sequence numbering: 15 to 19, 21, 22, 26 to 30 and 33, wherein, extra peptide is to select according to experimenter HLA group to be treated, has at least a kind of peptide to be selected from following one group of sequence numbering (optional): from survivin HLA-A1 sequence numbering: 38 BIR-002b FTELTLGEF, from survivin HLA-A2 sequence numbering: 39 BIR-002c LMLGEFLKL, from survivin HLA-B35 sequence numbering: 40 BIR-002d EPDLAQCFY, from survivin HLA-DR BIR-002a sequence numbering: 41 TLGEFLKLDRERAKD and from survivin HLA-DR and HLA-A*02BIR-004 sequence numbering: 42 ELTLGEFLKLDRERAKN, wherein, extra peptide is to select according to experimenter HLA group to be treated.

In another embodiment, compositions comprises 10 kinds of peptides that are comprised of following sequence numbering aminoacid sequence: sequence numbering: 20, 23 to 25, 31, 32, 1, 2, 5, 7, 9 to 11, 13, 14, have a kind of extra peptide of following sequence numbering at least: sequence numbering: 15 to 19, 21, 22, 26 to 30 and 33, wherein, extra peptide is according to there being the experimenter of needs HLA group to select, and having a kind of peptide at least is to be selected from following one group of sequence numbering (optional): from survivin HLA-A1 sequence numbering: 38 BIR-002b FTELTLGEF, from survivin HLA-A2 sequence numbering: 39 BIR-002c LMLGEFLKL, from survivin HLA-B35 sequence numbering: 40 BIR-002d EPDLAQCFY, from survivin HLA-DR BIR-002a sequence numbering: 41 TLGEFLKLDRERAKD and from survivin HLA-DR and HLA-A*02BIR-004 sequence numbering: 42 ELTLGEFLKLDRERAKN, wherein, extra peptide is to select according to experimenter HLA group to be treated.

WO2004/067023 has introduced the derivative MHC I class restriction peptide of tumor associated antigen survivin, and these Toplink are combined with HLA I quasi-molecule high-affinity.

The optimised quantity of the contained every kind of peptide of vaccine and best dosage regimen can determine those skilled in the art by one, and without carrying out undo experimentation.For example, peptide or its variant can be prepared as vein (i.v.) injection, subcutaneous (s.c.) injection, Intradermal (i.d.) injection, intraperitoneal (i.p.) injection, muscle (i.m.) injection.Peptide can, by subcutaneous, Intradermal, intraperitoneal, intramuscular and intravenous injection, be only the not ways to restrain of giving an example.DNA can be by Intradermal, intramuscular, subcutaneous, intraperitoneal and intravenous injection, is only ways to restrain not for example.May provide as peptide or the DNA dosage of 1 to 500mg, 50 μ g to 1.5mg or 125 μ g to 500 μ g, this depends on peptide or DNA separately.In the former test of above-mentioned dosage range, successfully use (Brunsvig PF, Aamdal S, Gjertsen MK, Kvalheim G, Markowski-Grimsrud CJ, Sve I, Dyrhaug M, Trachsel S,

eriksen JA, Gaudernack G; Telomerase peptide vaccination:a phase I/II study in patients with non-small cell lung cancer; Cancer Immunol Immunother.2006; 55 (12): 1553-1564; M.Staehler, A.Stenzl, P.Y.Dietrich, T.Eisen, A.Haferkamp, J.Beck, A.Mayer, S.Walter, H.Singh, J.Frisch, C.G.Stief; An open label study to evaluate the safety and immunogenicity of the peptide based cancer vaccine IMA901, ASCO meeting 2007; Summary numbering: 3017).

Compositions disclosed herein may collect, and wherein, thereby in compositions, the selection of peptide and contained quantity can be specifically for tissue, cancer and/or patients.For example, the accurate selection of the bootable peptide of expression pattern of parent's albumen in particular organization, thus avoid side effect.This selection may depend on the concrete cancer types of patient to be treated and morbid state, early treatment's scheme, patient's immune state, certainly, also has patient's HLA haplotype.And according to particular patient individual demand, vaccine of the present disclosure can contain individuation composition.According to the expression of the relevant TAA of particular patient, due to the adjustment to Retreatment or therapeutic scheme after individual is irritated or other Therapeutic Method produces unexpected side effect and first round treatment, the peptide content in each embodiment is different.

Technical staff can be by the preferred compositions of following test selection immunogenic peptide, as the production of the outer T cell of: test body and effect thereof and total amount, for propagation, affinity and the amplification of some T cell of some peptide, and test the functional of T cell by analyzing the generation (consulting below embodiment) of IFN-γ.Conventionally for above-mentioned object, then the most effective peptide is combined into a kind of vaccine.

Applicable vaccine may contain 2,3,4,5,6,7,8,9,10,11,12,13,14,15,16,17,18,19 or 20 kind of different peptide.Peptide length for cancer vaccine can be the peptide of any appropriate.Particularly, it may be a kind of suitable 9-mer peptide or a kind of suitable 8-mer or 9-mer or 10-mer or 11-mer peptide or 12-mer, 13-mer, 14-mer or 15-mer peptide..Longer peptide also may be applicable to, and 9 peptides or 10 peptides in

subordinate list

1 and 2, described are first-selections of MHC I class peptide, and 12 to 15 peptides are first-selections of MHC II class peptide.

Peptide has formed tumor or cancer vaccine.This vaccine can directly be given to patient's influenced organ, also can whole body administration, or the external cell (again these cells being injected in patient subsequently) being applied to from patient or its cell strain, or external for always from the cell subsets (and then again giving patient by cell) of patient's immunocyte.

This peptide may be pure peptide, may be also with the compositions (seeing below) of immunostimulation adjuvant or combines with the immunostimulatory cell factor and use or for example, with suitable induction system administration, liposome.This peptide also can conjugation form a kind of suitable carrier (as keyhole-limpet hemocyanin (KLH) or manna) to suitable carrier (consulting (1993) Ann.NY Acad.Sci.690, the 276-291 such as WO95/18145 and Longenecker).This peptide also can carry out labelling or a kind of fusion rotein or hybrid molecule.The peptide that provides sequence is estimated to stimulate cd4 t cell or CD8CTL.But, while having reverse CD positive T cell to offer help, stimulate more effective.Therefore,, for the MHC-II class epi-position that stimulates cd4 t cell, a kind of fusion partners of hybrid molecule or fragment provide the suitable epi-position that stimulates CD8 positive T cell.On the other hand, for the MHC I class epi-position that stimulates CD8CTL, a kind of fusion partners of hybrid molecule or fragment provide the suitable epi-position that stimulates CD4 positive T cell.It is all well known in the art that CD4 and CD8 stimulate epi-position, and they comprise the epi-position that those are definite in the disclosure.

Acceptable pharmaceutical carrier be known and be generally liquid, can prepare a kind of effective therapeutic agent system wherein.Although the carrier in dosage form may have chemistry and/or biological stability, release characteristics etc., does not generally possess any pharmacologically active.Representative formula can be such as Alfonso R.Gennaro. < < Remington:The Science and Practice of Pharmacy > >, 20th Edition.Baltimore, MD:Lippincott Williams & Wilkins, in 2000 books such as grade, find, include, but is not limited to saline, water, buffer, 0.3% glycine, hyaluronic acid, glucose etc.Recently, it is found that in human patients, being used as some lipomul in a lot of years of intravenous nutrition also can serve as a kind of peptide excipient.There are two kinds of these type of Emulsions as commercial lipomul, are named as Intralipid (Intralipid) and Lipofundin (fat of trying hard to keep is peaceful).The U.S. 3,169, No. 094 patent describe " registered trade mark of Intralipid ”Shi Sweden Ka Bi drugmaker intravenous nutrition lipomul, " fat of trying hard to keep is peaceful " is the registered trade mark of German Braun Medical Inc..Both all comprise soybean oil fat (containing 100 or 200 grams in 1000 distilled water: content is respectively 10% or 20%).Intralipid is made emulsifying agent (12g/l distilled water) with egg yolk lecithin, and the fat of trying hard to keep is rather with Ovum Gallus domesticus Flavus lecithin (2g/l distilled water).In Intralipid and the fat of trying hard to keep are peaceful, add glycerol (25g/l) to produce isotonicity.

In order to trigger immunoreation, conventionally need to add to allow the compositions more can immunogenic adjuvant.On the other hand, the compositions that proposition comprises at least one HLA binding peptide and immunological adjuvant, wherein HLA binding peptide comprises the epi-position from prostate related antigen molecule.

In literary composition, term used " immunological adjuvant " refers to and combines with antigen molecule while using, any material of non-specific quickening, prolongation or enhancement antigen specific immune response.Immunological adjuvant is to know in affiliated field, and any immunological adjuvant can be used.Applicable adjuvant include but not limited to 1018ISS, aluminum salt,

aS15, BCG, CP-870,893, CpG7909, CyaA, Mologen ' s

gM-CSF, IC30, IC31,

imuFact IMP321, interferon-ALPHA or β, IS Patch, ISS, ISCOMs,

mF59, Monophosphoryl lipid A and other nontoxic LPS derivants, Montanide IMS1312, Montanide ISA206, Montanide ISA50V, Montanide ISA-51, OK-432, OM-174, OM-197-MP-EC, ONTAK,

carrier system, PLG microgranule,

sRL172, virion and other virus-like particles, YF-17D, VEGFR1R2-Fc DELTA C1(a), R848, beta glucan, Pam3Cys, from the QS21 stimulon (the A Kuila biotechnology company of Massachusetts, United States Wu Site) of the Aquila company of Saponin, Mycobacterium extract and synthetic bacteria cell wall analogies and other patent adjuvants as the Detox of Ribi company, Quil or

adjuvant as

incomplete Freund's adjuvant, interferon-' alpha ' or GM-CSF are preferred.Before more a pair of specific for dendritic cells immunological adjuvants (as MF59) and preparation method thereof be described (Dupuis M, MurphyTJ, Higgins D, Ugozzoli M, van Nest G, Ott G, McDonald DM; Dendritic cells internalize vaccine adjuvant after intramuscular injection; Cell Immunol.1998; 186 (1): 18-27; Allison AC; The mode of action of immunological adjuvants; Dev Biol Stand.1998; 92:3-11).Also may use cytokine.Some cytokines directly affect dendritic cell to lymphoid tissue migration (as; TNF-Ru); accelerate maturing dendritic cell and be the lymphocytic Effective Antigens presenting cells of T (as, GM-CSF, IL-1 and IL-4) (the U.S. 5,849; No. 589 patents; especially the complete form with its list of references is incorporated to herein), and serve as immunological adjuvant (as, IL-12) (Gabrilovich DI; Cunningham HT, Carbone DP; IL-12and mutant P53peptide-pulsed dendritic cells for the specific immunotherapy of cancer; J Immunother Emphasis Tumor Immunol.1996 (6): 414-418).

CpG immunostimulatory oligonucleotide is also in the news and can strengthens the effect of adjuvant, itself is the adjuvant as vaccine.The constraint of gear shaper without theoretical, CpG ODN can by Toll sample receptor (TLR) (being mainly TLR9) activate congenital (non-habitual) thus immune system works.The TLR9 activation that CpG causes has improved antigenic specificity body fluid and the cell effect to various antigens, and these antigens comprise the polysaccharide conjugate in peptide or proteantigen, live virus or killed virus, dendritic cell vaccine, autogenous cell vaccine and preventative and therapeutic vaccine.The more important thing is, it can strengthen the ripe and differentiation of dendritic cell, causes T _h1the activation enhancing of cell and cytotoxic T lymphocyte (CTL) generate to be strengthened, and is not even having under cd4 t cell help.Even when the existing of vaccine adjuvant, the T that TLR9 activation is brought out _h1skew also can maintain, these adjuvants as: normally promote T _h2alumen or the incomplete Freund's adjuvant (IFA) of skew.CpG ODN when preparation or administering drug combinations, shows stronger adjuvanticity together with following other adjuvants or formula, and as microgranule, nanoparticle, fat milk or similar formulations, when antigen is relatively weak, these are particularly necessary to bringing out strong reaction.They can also accelerate immunoreation, make antigen dose reduce by approximately two orders of magnitude, in some experiment, to not containing the full dosage vaccine of CpG, also can produce similar antibody response (Arthur M.Krieg, Therapeutic potential of Toll-like receptor9activation, Nature Reviews, Drug Discovery, 2006,5,471-484).The U.S. 6,406,705B1 patent is combined with and impels antigen specific immune reaction to be described CpG oligonucleotide, non-Nuclec acid adjuvants and antigen.Commercially available CpG TLR9 antagonist is that Mologen company (Berlin, Germany) produces

(two loop-stem structure immunomodulator).Also can use other as TLR binding molecule, as: RNA is in conjunction with TLR7, TLR8 and/or TLR9.

The CpGs(that other examples of useful adjuvant include but not limited to chemical modification is as CpR, Idera), Mucin1-mRNA/ protamine complex, poly-(I:C) (as poly-I:C12U), non-CpG DNA of bacteria or RNA and immunocompetence micromolecule and antibody are as imidazoquinolie, cyclophosphamide, Sutent, bevacizumab, celecoxib, NCX-4016, sldenafil, tadanafil, Vardenafil, Sorafenib, XL-999, CP-547632, pazopanib, ZD2171, AZD2171, easy Puli's nurse agate and SC58175, they may be used as treatment and/or adjuvant.Technical staff is just easy to determine adjuvant useful in the disclosure and quantity and the concentration of additive without undo experimentation.

In one embodiment, adjuvant system by

bCG, OK432, imiquimod, Mucin1-mRNA/ protamine mixture, resimiquimod, GM-CSF, interferon-' alpha ', with

or select in its compositions form a group.

In another embodiment, adjuvant system by colony stimulating factor as granulocyte macrophage colony stimulating factor (GM-CSF, Sargramostim),

in one group that Mucin1-mRNA/ protamine mixture, resimiquimod and interferon-' alpha ' form, select.

In another embodiment of disclosure compositions, adjuvant is Mucin1-mRNA/ protamine mixture, imiquimod or resimiquimod.

In literary composition, disclosed said composition can be used for parenteral, as subcutaneous, Intradermal, intramuscular, peritoneal injection or oral administration.For this reason, peptide and other selectivity molecules decompose or suspend in pharmaceutical carrier, are preferably water carrier.In addition, compositions can comprise adjuvant, as: buffer agent, bonding agent, impact agent, diluent, spice, lubricant etc.These peptides also can share with immunostimulation material, as: cytokine.The more adjuvants that can be used for such composition can be the < < Handbook of Pharmaceutical Excipients > > showing from A.Kibbe (the 3rd edition, 2000 Nian， American Medical Associations and pharmacy publishing house) etc. know in book.This composition of medicine can be used for stoping, preventing and/or treating adenoma or Cancerous disease, is preferably colorectal cancer.

Cytotoxic T cell (CTL) can identify with MHC molecule rather than with the antigen occurring with a kind of peptide form of the combination of complete exotic antigen own.MHC molecule itself is positioned at the cell surface of antigen presenting cell.Therefore,, while only occurring peptide antigen, MHC molecule and APC trimer compositions, just may activate CTL.Correspondingly, if not only have this peptide for activating CTL, but add the extra APC containing each MHC molecule, can improve immunoreation.

In another embodiment, compositions of the present disclosure also contains at least one antigen presenting cell.

Antigen presenting cell (or stimulating factor cell) has a MHC I class or II quasi-molecule conventionally on its surface, in one embodiment, this antigen presenting cell self can not be written into selected antigen to MHC I class or II quasi-molecule substantially.Described in detail as follows, MHC I or II quasi-molecule in vitro may the easy selected antigens of load.

In one embodiment, mammalian cell lacks or reduces level or the function of TAP peptide transporter.The applicable cell that lacks TAP peptide transport vehicle comprises that human body peptide is written into the catalog number (Cat.No.) CRL1992 of deficient cells ZhuT2， subordinate American Type Culture Collecti (12301Parklawn Drive, Rockville, Maryland20852, the U.S.); TAP deficient cells strain such as T2 can be used as APC, and owing to lacking TAP, therefore nearly all peptide of being offered by MHC I quasi-molecule all receives publicity, and is written into the unloaded MHC-I quasi-molecule of these cell strains for outside, and all effects are all by far and away owing to the peptide using.

In another embodiment, antigen presenting cell is dendritic cell.Suitable dendritic cell is autologous dendritic cell, makes impulse process by antigenic peptides.Antigenic peptides may be any applicable antigenic peptides that causes suitable t cell responses.The people such as Murphy (1996) are at < < prostate > > magazine (The Prostate29,371-380 and The Prostate32,272-278), disclosed a kind of T cell therapy of using autologous dendritic cell, autologous dendritic cell carries out impulse processing by a kind of peptide of antigen related neoplasms.

In another embodiment, peptide impulse or load peptide for the compositions that contains at least one antigen presenting cell.

As a kind of substitute, antigen presenting cell comprises the expression vector of an encoded peptide.Polynucleotide can be any applicable polynucleotide.In one embodiment, polynucleotide can dendritic cells, thereby causes offering and immunity induction of peptide.

Applicable, nucleotide of the present disclosure may be comprised in viral polynucleotide or virus.For example, the dendritic cell of adenoviral transduction shows to induce the antigenic specificity antineoplastic immune relevant to MUC1 (to consult Gong et al (1997) Gene Ther.4,1023-1028). similarly, can use the system based on adenovirus (for example to consult, Wan et al (1997) Hum.Gene Ther.8,1355-1363); Can use retrovirus system (Specht et al (1997) J.Exp.Med.186,1213-1221 and Szabolcs et al (1997) (also can be used blood particle mediation to be passed to the method (Tuting et al (1997) Eur.J.Immunol.27,2702-2707) of dendritic cell; Also can use RNA (Ashley et al (1997) J.Exp.Med.186,11771182).

In general, in the disclosure, the disclosure compositions that contains a kind of (multiple) nucleic acid can the mode similar to those compositionss that contain disclosure peptide be used, as in vein, tremulous pulse, abdominal cavity, muscle, Intradermal, tumor, oral cavity, percutaneous, via intranasal application, buccal, per rectum, suction or external.

Due to the mechanism of escaping, tumor often produces drug resistance to medicine.Drug resistance may occur, and show in transfer and recurrent tumor during treating.For fear of this drug resistance, tumor is treated by administering drug combinations conventionally, after the tumor of metastatic tumor and the recurrence of anosis after date, often needs different drug regimens to treat.Therefore,, in one side of the present disclosure, compositions is combined use with the second anticarcinogen.The second medicine may be used in the front and back of disclosure compositions or simultaneously.For example, if chemical property is compatible, compositions of the present disclosure is mixed with the second anticarcinogen can realize administration simultaneously.The another kind of mode of simultaneously administration is that compositions and anticarcinogen are being used different route of administration administrations on the same day, and for example compositions may be to be injected and the second anticarcinogen is oral.Compositions and the second anticarcinogen also may be used in the same course for the treatment of, but are not to use on the same day and/or in the different courses for the treatment of.

In another embodiment, proposed the method for the treatment of or prevention patient cancer, the method comprises any current disclosed compositions of using treatment effective dose to patient.

Effectively therapeutic dose refers to and can cause immunoreactive amount, particularly activates CTL subgroup.One of ordinary skill in the art, by using standard immunoassay method as those methods that provide in this description embodiment, can be easy to determine that whether dosage is effective.Another method of monitoring certain dose-effect fruit of this compositions is to observe growth and/or its recurrence of being treated tumor.

In another embodiment, compositions is used as anti-cancer vaccine.

Nucleic acid containing peptide combinations or peptide coding also can form the vaccine of tumor or cancer.This vaccine can directly be given to patient's influenced organ, also can whole body administration, or the external cell (again these cells being injected in patient subsequently) being applied to from patient or its cell strain, or external for always from the cell subsets (and then again giving patient by cell) of patient's immunocyte.

On the one hand, vaccine is the Multiple Peptide tumor vaccine that is used for the treatment of carcinoma of prostate.On the other hand, vaccine comprises the one group of tumor-associated peptides that is selected from following sequence numbering: sequence numbering: 1 to 11 and sequence numbering: 13 to 14, they are arranged in and find at former prostatic cell and/or carcinoma of prostate.This group comprises HLA I class and II class peptide.These peptides also can at least comprise a kind of peptide in influenza cAg, and this peptide is that positivity is controlled peptide, is the immune marker that detects intradermal administration curative effect.In another particular, vaccine forms (its sequence numbering is 1 to 14) by 14 kinds of individual peptides, wherein the content of every kind of peptide is to select in one group that is comprised of to approximately 75 μ g, approximately 1000 μ g to approximately 175 μ g, approximately 500 μ g to approximately 600 μ g and approximately 578 μ g approximately 1500 μ g, all peptides may be purified with HPLC and ion exchange chromatography, are shown as white to off-white powder.Lyophilized products can be dissolved in sodium bicarbonate, at room temperature recombinates in latter 30 minutes for intradermal injection.The peptide total amount of every 500 μ l solution can not be approximately 0.1 to 100mg, approximately 0.1 to 1mg and approximately 300 μ g to 800 μ g not etc.If do not make different statements, term herein " approximately " refers to the average +/-10% of set-point.Technical staff can adjust the actual content of peptide to be used as the amount of TUMAP in the immune state of individual patient and/or specific types of cancer according to several factors.Peptide may replace dried frozen aquatic products to provide in other suitable modes (sterile solution etc.).

The peptide that compositions may comprise free form or exist with a kind of pharmaceutical salts form.

" pharmaceutical salts " used herein means a kind of derivant of disclosed peptide, and wherein this peptide carries out modification by the alkali salt of antacid or medicament.For example, use that (neutral medicine wherein has a Zhong – NH conventionally with the free alkali of applicable acid reaction ₂group) prepare acid salt.The acid that is applicable to preparing hydrochlorate comprises organic acid, as: acetic acid, propanoic acid, hydroxy acid, acetone acid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzic acid, cinnamic acid, mandelic acid, methanesulfonic acid, methanesulfonic acid, benzenesulfonic acid, salicylic acid etc. and mineral acid, as: hydrochloric acid, hydrobromic acid, sulphuric acid, nitric acid and phosphoric acid etc.On the contrary, the alkali salt preparation of the acidic-group that can offer on a kind of peptide is used medicinal base to be prepared, as sodium hydroxide, potassium hydroxide, ammonium hydroxide, calcium hydroxide, trimethylamine etc.Compositions may comprise the peptide of acetic acid (acetate), ammonium or hydrochloric acid (chloride) salt form, only for being not limited to for example these.

In one embodiment, compositions may comprise that sugar, sugar alcohol, aminoacid (as: glycine, arginine, glutamic acid etc.) are as skeleton forming agent.These sugar may be monosaccharide and disaccharide or trisaccharide.These sugar can be used alone, and also can combine with sugar alcohol.The example of sugar comprises the melitriose of glucose, mannose, galactose, fructose or the sorbose of monosaccharide, sucrose, lactose, maltose or the trehalose of disaccharides and three saccharides.Sugar alcohol may be, for example, and mannose.In one embodiment, compositions comprises sucrose, lactose, maltose, trehalose, mannitol and/or sorbitol, and in another embodiment, compositions comprises mannitol.

In addition, compositions may comprise that adjuvant that physiological tolerance is good is referring to < < pharmaceutic adjuvant handbook > > (Handbook of Pharmaceutical Excipients), the 5th edition, Raymond Rowe, Paul Sheskey and Sian Owen write, medical publishing house (2006)), as antioxidant, as ascorbic acid or glutathion, antiseptic is as phenol, m-cresol, methyl hydroxybenzoate or propyl ester, methaform, thimerosal or benzalkonium chloride, stabilizing agent, framework forms agent as sucrose, lactose, maltose, trehalose, mannose, mannitol and/or sorbitol, mannitol and/or lactose and solubilizing agent are as Polyethylene Glycol (PEG), i.e. PEG3000, PEG3350, PEG4000 or PEG6000 or cyclodextrin, i.e. hydroxy propyl-Beta-cyclodextrin, fourth second sulfo--cycloheptaamylose or γ cyclodextrin, or dextran or poloxamer, i.e. poloxamer188, PLURONICS F87 or

20, 80.On the other hand, may comprise that one or more are selected from the good adjuvant of toleration of the group of antioxidant, skeleton forming agent and stabilizing agent composition.

On the other hand, the pH of vein and intramuscular injection is selected from pH2 to pH12, and hypodermic pH is selected from pH2.7 to pH9.0, because dilution rate reduces in body, causes more may spreading in injection site.Strickley?Robert?G.,Pharm.Res.,21,NO:2,201-230(2004)。

On the other hand, use the pharmaceutical preparation that comprises disclosure peptide and/or nucleotide to each peptide or relevant adenoma or the Cancerous disease patient of antigen.Can trigger by this method the cell-mediated immunoreation of T.

The present invention further discloses a kind of compositions, the quantity of peptide of the present disclosure that wherein compositions contains (especially Tumor-assaciated), nucleotide or expression vector is tissue, cancer and/or patient-specific.

In another embodiment, vaccine is nucleotide vaccine.As everyone knows, inoculation nucleic acid vaccine (as DNA vaccination) coded polypeptide can cause t cell responses.This vaccine can directly be given to patient's influenced organ, also can whole body administration, or the external cell (again these cells being injected in patient subsequently) being applied to from patient or its cell strain, or external for always from the cell subsets (and then again giving patient by cell) of patient's immunocyte.If nucleic acid gives cell in vitro, may be of value to cell transfecting, with the co expression immunostimulatory cell factor, as interleukin II or GM-CSF.Nucleic acid can be completely individually dosed, also can combine use with immunostimulation adjuvant or with the immunostimulatory cell factor, or for example, with suitable induction system administration, liposome.Nucleic acid vaccine can be used together with adjuvant, the adjuvant using together with peptide vaccine as above-mentioned.Nucleotide vaccine can not used together with adjuvant.

Polynucleotide can be basic purified form, also can be coated in carrier or induction system.Suitable carrier and induction system comprise viral system, as based on adenovirus, vaccinia virus, retrovirus, herpesvirus, adeno-associated virus or containing the system of the hybrid virus of more than one viral elements.Non-viral induction system comprises cationic-liposome and cationic polymer, is all the method for knowing in DNA delivery value territory.Also can use physical transport system, as passed through " particle gun ".The peptide of peptide or nucleic acid coding can be a kind of fusion rotein, for example, and containing an epi-position (stimulating CD-4 positive T cell) of tetanus toxoid.

Suitable situation is that any nucleic acid that gives patient is all aseptic, pyrogen-free.Naked DNA can intramuscular injection, intradermal injection or subcutaneous injection.Talk about aptly, nucleotide vaccine may comprise any suitable nucleotide means of delivery.Nucleic acid is available a kind of liposome or the conveying of viral vector induction system also.In one embodiment, nucleotide vaccine is injected in muscle as DNA vaccination.In another embodiment, peptide vaccine is by subcutaneous or intradermal injection.In another embodiment, vaccine is injected in skin.

We think, the expression of the peptide of the absorption of nucleic acid and professional antigen presenting cell (as dendritic cell) coding may be due to immunoreactive Initiated Mechanism, yet, dendritic cell is may not can transfected, but because they may pick out the peptide being expressed in the transfectional cell from tissue, therefore still very important (" intersect and start ", for example, Thomas AM, Santarsiero LM, Lutz ER, Armstrong TD, Chen YC, Huang LQ, Laheru DA, Goggins M, Hruban RH, Jaffee EM.Mesothelin-specific CD8 (+) T cell responses provide evidence of in vivo cross-priming by antigen-presenting cells in vaccinated pancreatic cancer patients.J Exp Med.2004Aug2, 200 (3): 297-306).

Conry etc. (1996) are at < < oncology collection of thesis > > (Seminars in Oncology23, 135-147), Condon etc. (1996) are at < < nature-medical science > > (Nature Medicine2, 1122-1127), Gong etc. (1997) are at < < nature-medical science > > (Nature Medicine3, 558-561), Zhai etc. (1996) are at < < Journal of Immunology > > (J.Immunol.156, 700-710), Graham etc. (1996) are at < < international journal of cancer > > (Int J.Cancer65, 664-670), Burchell etc. (1996) 309-313 is at Breast Cancer, in Advances in biology and therapeutics mono-literary composition, Calvo etc. (Eds), all to polymerized nucleoside, acid mediated cancer immunityization treatment is described John Libbey Eurotext, with complete reform, be incorporated to herein.

This for example also may be of value to vaccine specific targeting, in cell mass (antigen presenting cell), can use local injection, use targeting vector and induction system, or for example, to the selective purification of such cell mass of patient and externally (give peptide or nucleic acid, dendritic cell can be by the people such as Zhou (nineteen ninety-five) at < < blood > > (Blood86, 3295-3301), the people such as Roth (1996) are at < < Scand.J.Immunology > > 43, method described in 646-651 is classified).For example, targeting vector can comprise a tissue or tumor-specific promoters, and guiding antigen is expressed in position.

Vaccine may depend on the particular type of patient's cancer to be treated and the state of an illness, early treatment's scheme, patient's immune state, certainly also has patient's HLA haplotype.And according to particular patient individual demand, vaccine can contain individuation composition.According to the expression of the relevant TAA of particular patient, due to the adjustment to Retreatment or therapeutic scheme after individual is irritated or other Therapeutic Method produces unexpected side effect and first round treatment, the peptide content in each embodiment is different.

Except useful to treatment cancer, in literary composition, disclosed peptide is also useful to diagnostics.Because a lot of peptides are produced and determined that these peptides do not exist in normal structure by carcinoma of prostate, so these peptides can be used for the existence of cancer diagnosis.

The peptide being present in biopsy can help pathologist cancer diagnosis.By antibody, mass spectrograph or other method known in the art, detecting some disclosed peptide can make pathologist judge that this is organized as pernicious, inflammation or general pathological changes.The existence of current disclosed peptide group can be to pathological tissues classification or segmentation.

The detection of pathological changes specimen being carried out to disclosure peptide makes to judge the interests of immune system Therapeutic Method, if particularly T lymphocyte is known or expectation is relevant with mechanism of action.The disappearance that MHC expresses is a kind of mechanism, has absolutely proved which infected malignant cell escaped immune surveillance.Therefore, the existence of current open peptide shows that this mechanism do not utilized by cell that analyzed.

Perhaps, current open peptide is used to analyze the lymphocyte reaction of those peptides, as t cell responses or the antibody response of peptide or MHC molecule complex peptides.These lymphocyte reactions can be used as prognostic indicator, determine whether to take further treatment.These reactions also can be intended to induction of lymphocyte reaction by different way as the Substitute Indexes in immunotherapy, as inoculation protein vaccine, nucleic acid, autologous material, lymphocyte adoptive transfer.In gene therapy, the lymphocyte reaction of current open peptide can be considered in side effect assessment.Lymphocyte reaction monitoring also may become a kind of valuable instrument in transplantation therapy follow-up examination, as, for detection of graft-versus-host and host versus graft disease.

On the one hand, the disclosure relates to an external member, comprises (a) container, the compositions that comprises above-mentioned solution or lyophilized powder form again; (b) option, second container, containing diluent or the restructuring solution of lyophilized powder dosage form; And (c) option, (i) use solution or (ii) restructuring and/or use the description of lyophilized powder dosage form.This external member also walks and comprises one or more (iii) buffer agent, (iv) diluent, (v) filtrate, (vi) syringe needle, or (v) syringe.In one embodiment, container is to select from following one group: bottle, cillin bottle, syringe, test tube or multipurpose container.In another embodiment, compositions is dried frozen aquatic products.

On the one hand, external member may comprise lyophilized formulations and restructuring and/or the operation instructions of current disclosed compositions in fitted vessel and/or vaccine.Suitable container comprises, for example bottle, cillin bottle (as double chamber bottle), syringe (as double-chamber syringe) and test tube.This container may be made of a variety of materials, as glass or plastics.In one embodiment, external member and/or container contain container or about the description of container, the explanation that indicates restructuring and/or use.For example, label may show that freeze-dried formulation will be reassembled as above-mentioned peptide concentration.This label can further show that preparation is for subcutaneous injection.

The container of depositing preparation can use multipurpose cillin bottle, makes can repeat to give (for example, 2-6 time) restructuring dosage form.This external member can further comprise second container that suitable diluents (as sodium bicarbonate solution) is housed.

In one embodiment, when mixed diluent and lyophilized formulations, the final peptide concentration of recombination preparation is at least 0.15mg/mL/ peptide (=75 μ g), is no more than 3mg/mL/ peptide (=1500 μ g).This external member also can comprise other material that business and user perspective are desirable, comprises other buffer agent, diluent, filtrate, syringe needle, syringe and with the package insert of operation instructions.

External member may have an independent container, wherein comprises composite preparation, and said preparation can have other composition (for example, other compound or and pharmaceutical composition), also can be without other composition, or every kind of composition has its different vessels.

In addition, external member may comprise the current packaged preparation that discloses compositions and/or vaccine of combining use with the second compound (as adjuvant (as imiquimod), chemotherapeutic, natural product, hormone or antagonist, anti-angiogenic agent or inhibitor, inducer of apoptosis or chelating agen).The composition of this external member can combine in advance or every kind of composition can be positioned over independent different vessels before giving patient.The composition of test kit may be provided by one or more liquid solution.In one embodiment, liquid solution is aqueous solution.In another embodiment, liquid solution is aseptic aqueous solution.The assembly of external member may be also solid form, and by the appropriate solvent that may be provided by other different vessels is provided, it can be transformed into liquid.

The container for the treatment of external member may be the instrument of cillin bottle, test tube, flask, bottle, syringe or any other splendid attire solid or liquid.Conventionally, when composition is not only a kind of, external member will comprise second cillin bottle or other container, and making it can be quantitative separately.This external member also may comprise another container that loads medicinal fluid.In one embodiment, treatment external member will comprise a device (as, one or more syringe needles, syringe, eye drop device, pipette etc.), medicine of the present disclosure (compositions of this external member) can be injected.

Pharmaceutical preparation may be to be applicable to any a kind of as mouth (enteral), nose, eye, subcutaneous, Intradermal, intramuscular, intravenous or percutaneous of route of administration that accept.On the one hand, route of administration is subcutaneous, can pass through infusion pump administration.

Term " t cell responses " refers to proliferated specifically and the activation of the effector function of induction in the external or body of peptide.For MHC I class restriction CTL, effector function may be dissolving, cytokine secretion, effector molecule secretion or the threshing that peptide impulse, peptide precursor impulse or native peptides are offered target cell.In one embodiment, t cell responses is the cytokine secretion of inducing peptide, and wherein, peptide selects free interferon-γ, TNF-α or IL-2 composition one group.In one embodiment, t cell responses is the effector molecule secretion of inducing peptide, and wherein, effector molecule selects free granzyme and perforin composition one group.For MHC II class restricted T accessory cell, effector function may be the cytokine secretion of inducing peptide, includes but not limited to the threshing of IFN-γ, TNF-α, IL-4, IL5, IL-10, IL-2 or inducing peptide.The possible effector function of CTL and helper T lymphocyte is not limited only to function described in this list.

The combination that comprises the peptide ammino acid sequence that can combine with major histocompatibility complex (MHC) MHC I class (HLA I class) or II class (HLA II class) is also disclosed.

Compositions can be used as the effectively vaccine against prostate cancer based on peptide combination.

Be the method for the treatment of carcinoma of prostate on the other hand, comprise and give disclosed any compositions in prostate patient literary composition.These compositionss can be used or use separately together with immunological adjuvant treatment.If use immunological adjuvant, it can comprise in disclosed in the text compositions or be individually dosed with identical or different route of administration.

Compositions disclosed herein and method can be used as starting treatment, auxiliary treatment or palliative therapy, also can be used alone or include but not limited to that with other treatment operative treatment (comprising prostatectomy), radiation and chemotherapy combine use.Disclosed method also can be used for primary tumor reaction, recurrence biology, local recurrence or transfer and relapse.Current disclosed method also can be deprived the forward and backward use for the treatment of or treat associating use with androgen-deprivation in androgen responsive type carcinoma of prostate and androgen independent form prostatic csarcinoma androgen.

Term used " androgen responsive type carcinoma of prostate " refers to that tumor growth needs androgenic any carcinoma of prostate herein.

Any carcinoma of prostate of tumor growth when term used " androgen independent form carcinoma of prostate " refers to without androgen herein.

Term used " androgen-deprivation treatment " refers to mainly contain and suppresses any treatment that androgen signal conducts or androgen tells on herein.

Following examples are only for illustrating that some aspect is not intended to limit the disclosure.All lists of references of quoting herein are all intactly incorporated herein with the form of quoting.

Embodiment

embodiment 1

Detected the HLA binding peptide mixture of biochemical patients with recurrent show to have the radical prostatectomy that shifts performance without diagnostic evidence after, determined whether said composition is effective aspect stable and/and increase PSA doubling time (" DT ").

patient selects

All patients have biochemical recurrence after the preliminary curative therapy by radical prostatectomy.Recurrence biology is defined as the measuring method assessment of being separated by least 14 days with two kinds, and the floor level from postoperative or operation and radiotherapy is increased to the PSA value higher than 50%.Other criterion of acceptability comprise: through bone scan, the video picture of axle position and CT, determine non-metastatic disease or Local neoplasm recurrence, east tumor cooperative groups behavior state scoring is 0 or 1, age >45 year and <80 year, without corticoid or other immune modulating treatments; Without following radiotherapy, hormone or chemotherapeutic treatment; , epilepsy pernicious without other or pulmonary disease.Entering anthology when research, all patients are androgen responsive types, and they have stopped androgen-deprivation and treat and have 12 months at least before selected.All patients are that HLA-A*02 is positive.Patient characteristic is as shown in table 1 below.

Table 1

treatment

For preventing that tumor from escaping by gene mutation and antigenic deletion, use for the multivalence peptide combinations of specific T-cells widely.Compositions comprises 13 kinds of prostate related antigen molecule HLA I classes and the paraspecific synthetic HLA binding peptide of II, for activating cytotoxic CD8+ and CD4+T accessory cell.11 kinds of peptide (sequence numberings: 1 to 11) comprise HLA-A*0201 restriction epi-position.2 kinds of peptide (sequence numberings: 13 to 14) comprise HLA II class in conjunction with epi-position.The extra peptide that comprises an influenza virus HLA-A*0201 epi-position (sequence numbering: 12) be added into as the labelling peptide that activates the anamnesis reaction of CD8+T cell.500ml montanide ISA-51 emulsifying for peptide, and the dosage of the single peptide of subcutaneous injection 300mg/.

In addition, patient is randomized acceptance (1) immunological adjuvant, (2) high temperature or (3) without immunological adjuvant or high temperature.The immunological adjuvant using comprise (1) imiquimod (Medarex Inc. of Bad Homburg, Germany ), the Bayer Healthcare of (2) GM-CSF(Leverkusen, Germany

), (3) Mucin1-mRNA/ protamine complex (is shown in Scheel etc.; Therapeutic anti-tumor immunity triggered by injections of immunostimulating single-stranded RNA, Eur.J.Immunology 2006; 36 (10): 2807 – 2816).

3 patients (1,2 and No. 5 patient) do not accept immunological adjuvant or hyperthermia treatment.

4 patients (16,17,18 and No. 19 patients) accept Mucin1-mRNA/ protamine complex as immunological adjuvant.For the patient who accepts Mucin1-mRNA/ protamine complex, peptide and montanide ISA-51 emulsifying for 110 μ g immunological adjuvants.All the components is once injected.

4 patients (3,7,8 and No. 11 patients) accept imiquimod as immunological adjuvant.For these patients, after peptide treatment sealing, imiquimod cream external application in injection site.Indication patient water after 8 hours cleans imiquimod drug of topical application position.

6 patients (4,6,9,12,14 and No. 15 patients) accept GM-CSF as immunological adjuvant.For these patients, the immunological adjuvant of 225 μ g dosage is in the close subcutaneous injection in place, injection site.

2 patients (10 and No. 13 patients) accept hyperthermia treatment.For these patients, after treatment directly at injection site 20cm ²exposure skin of abdomen place use and to maintain the thermal source of 41 ° of C, and stop 20 minutes.

After preliminary treatment the 7th, 14,28,42 and 56 days, repeat peptide combinations, immunological adjuvant and/or hyperthermia treatment at the same position of each medication.Afterwards, if PSA value objectivity returns or be stable, continue within every 4 weeks, to treat until the 420th day.

reaction assessment

With each treatment, pay a return visit definite alternate parameter PSA measured value assessment therapeutic response.After initial 6 treatments, behind (at the 8th week) and thereafter every 3 months, repeat hematology and blood chemical examination.With assessment clinical progress similar timetable in carry out Clinical Laboratory and digital rectal examination.

data analysis

By calculating the geometrical mean of doubling time, assess reaction.The logarithm of every patient PSA value meets the straight line of variable slope.With nonlinear fitting program method of least square, estimate switching point, Different Slope and the initial value between two slopes.Complete reaction is defined as PSA value and can not surveys; Partial reaction is defined as PSA value and declines 50%; Stable disease is defined as PSA value and declines and to be no more than 50% or increase and be no more than 10%; Disease progression is defined as allowing benchmark PSA value increase more than 10% arbitrarily.These measured values must be verified after 4 weeks.To stopping, research patient's biochemical reaction is followed the trail of until they accept further local radiation or androgen-deprivation treatment.

result

The mean P SA doubling time (DT) before all patient treatments is 8.4 months, and treatment increases to 11.2 months while finishing.4 (21%) patients demonstrations obtain biochemical interests from treatment.In two patients that increase progressively in PSA value, with digital rectal examination, clinical tumor recurrence detected and confirm through PET-CT scanning.Therapeutic response and PSA value are different because of patient, can be divided into 5 different reaction groups.Data are organized in Fig. 2.

PSA stability and DT increase.Two patient (3 and No. 8 patients; 11%) be presented at 14 and 16 months period P SA stable (Fig. 3) following up a case by regular visits to after treatment and last medication.From begin treatment, stability is 29.5 months of data truncation value place lasting average time, has on average carried out 17 treatments (14 and 20 times).No. 3 patient has part PSA reaction (>50%), and the time is 9 months, is that PSA slowly increases the phase afterwards, compares with the treatment doubling time of first 9.8 months, and its doubling time is 20.5 months.PSA first before research occurs in postoperative 18 months of prostate excision (pT2pN0GS5).Anaphylaxis during due to the 20th treatment of No. 3 patients, he must exit research.No. 8 patients show that treatment starts rear illness behavior and stablizes.He stops treatment is because the anaphylaxis of the 14th treatment after 10 months.This patient suffers from pT3b Gleason 3+4 level tumor, and after radical prostatectomy, PSA minimum is 0.6ng/ml, and he includes in before research after declining first after surgery, shows that PSA increases.The DT that calculates him increased to 148.0 months from 6.6 months.The imiquimod that two patients accept percutaneous is as immunological adjuvant (Fig. 4).

pSA DT increases, and PSA is unstable.two patients (11 and No. 16 patients, 11%) increase at treatment period P SA DT, and follow PSA value slowly to increase.At first 6 months for the treatment of, No. 11 patients' PSA DT increased to 10.1 months by 1.5 months.This patient is begin treatment when PSA value is 10.8ng/ml, and after 6 months, increases to 17.8ng/ml in treatment.Research is terminated, and starts hormone replacement therapy.In this patient, with PET-CT scanning, do not find obvious malignant change.Use imiquimod as immunological adjuvant.No. 16 patients' DT is 6.1 months when research starts.At first 5 months for the treatment of, his PSA value declines and DT is become to the half-life was 2.7 months.Afterwards, the DT that statistical computation goes out him increases to 14.4 months, and has continued 16 months after treatment starts.When treatment starts, his PSA value is 0.29ng/ml; Following up a case by regular visits to while finishing, is 0.41ng/ml.He accepts Mucin1-mRNA/ protamine complex as immunological adjuvant (Fig. 3 and Fig. 4).

the interim PSA that PSA declines and PSA DT increases before raises.a patient (No. 5 patients, 5%) PSA after treatment starts increases unaffected, and he uses after the 11st kind of peptide during for 1.31ng/ml in PSA value, stops treating.Afterwards, his PSA reduces, and DT increases to 20.2 months until follow up a case by regular visits to end; Patient does not accept any other treatment during this.This patient does not use immunological adjuvant, only emulsifying in montanide (Fig. 3 and Fig. 4) of peptide.

the interim PSA that PSA accelerates to raise before declines or is stable.3 patients' (7,15 and No. 17 patients, 16%) PSA value keeps stable or declines, and is then then to accelerate to raise.No. 7 patients' DT is 3.7 months when treatment starts, and increases to 21.5 months, and continued 4 months during treating, and proceeds to afterwards 2.8 months.This patient suffers from pT2b tumor, and histology is that surgical resection margins is positive.He refuses any local radiotherapy (Fig. 3).No. 15 patients' DT is 1.3 months when research starts.Before treatment first, our hospital has determined latter two continuous PSA value between 4 months.Due to difference between laboratory, PSA DT result is from 1.3 monthly variation to 25.8 month.During treatment, combine and use GM-CSF, DT reduces to 9.9 months, and keeps stable at 6 months.Then, again to proceed to DT be 7.4 months to PSA.Before the explanation of this psa process is treated and starts, the short term variations of benchmark PSA DT hinders.In the half-life, be in the treatment phase during the interim PSA of 1.9 months treatment after declining, No. 17 patients' PSA DT reduced to 4.8 months from 10.2 months, and had continued 2 months, and PSA increases (Fig. 3) afterwards.

pSA progress.

During studying, 11 patients' (58%) PSA value progress is unaffected, and PSADT is constant, and research is by premature termination (Fig. 4).Use the meansigma methods of 13 times (scope is 7-18 time) treatment.

the impact of immunological adjuvant

Surpass 8 patients and show that PSA DT increases or PSA value declines, accept imiquimod as immunological adjuvant for 4.A patient accepts GM-CSF, and 2 patients accept Mucin1-mRNA/ protamine complex, and a patient does not use immunological adjuvant.In accepting two patients of localized hyperthermia's treatment, nobody responds or clinical benefit (Fig. 4).

embodiment 2

Its reactivity of specificity HLA I class binding peptide being included in mixtures of polypeptides detects after the treatment of embodiment 1.

Specific T-cells amplification in vitro

From the peripheral blood lymphocytes of carcinoma of prostate during inoculation different time point obtain, and be frozen in the liquid nitrogen containing 90% hyclone and 10%DMSO.After thawing, approximately 5 * 10 ⁶individual cell culture (24 porocyte culture plates, Greiner Bio-One company, Frickenhausen, Germany) in the IMDM culture medium, 50 μ g/ml streptomycins (all Biowhittaker company products that are that supplement 50U/ml penicillin, Verviers, Belgium), the beta-mercaptoethanol under 10% hot deactivation human serum (c.c.pro, Neustadt, Germany) and 50 μ M37 ° C condition and 7.5% CO ₂.At first day, add respectively HLA I class or the synthetic binding peptide of HLA II class of mixing, HLA I class 1 μ g/ml, HLA II class 5 μ g/ml.The HLA I class culture of giving for the 3rd, 5,7 and 9 days at T cytositimulation supplements recombinant human il-2 (r-hIL2 of Wiesbaden, Germany R & D Systems GmbH company), at the 0th day supplementary promokine and IL-4 and IL-7, at the HLA II class culture of giving for the 3rd, 5,7 and 9 days of T cytositimulation, supplement recombinant human il-2 (r-hIL2 of Wiesbaden, Germany R & D Systems GmbH company) and promokine.

Enzyme-linked Immunosorbent Assay speckle (ELISPOT) is measured

According to cancer immunotherapy association (CIP) immune guiding programmatic recommendation, the function of check amplification T cell in standard interferon-γ ELISPOT measures.In simple terms, cell is collected after within 12 days, cultivating, washing in the culture medium on ELISPOT plate (Millipore company, Schwalbach, Germany), counting and inoculation.There is i) peptide presenting cells is K562-A2 and every kind of HLA I class binding peptide of 1 μ g/ml or ii) every kind of HLA II class binding peptide of 2.5 μ g/ml in the situation that, detects two parts or three parts of 0.20x10 ⁶to 0.40x10 ⁶individual cell.PHA(10 μ g/ml) or SEB(1 μ g/ml) be used as positive control and stimulate.At 37 ° of C and 7.5%CO ₂condition under hatch after 26 hours, with a pair of monoclonal antibody specific (1D1-k and the 7-B6-1 of Sweden Nacka Strand Mabtech company), detect IFN-γ output.Add respectively ExtraAvidin-alkali phosphatase and BCIP/NBT substrate (both are the product of Sigma-Aldrich company) 1 hour 10 minutes.Use enzyme connection spot-analysis instrument (3A of Arlen, Germany CT company and 5 series) to carry out ELISPOT analysis.

For every kind of peptide of every patient, record produces the T cell of IFN γ and makes table.As shown in figure 13, in 11 kinds of HLA I class peptides, there is at least one patient's reaction of 8 kinds of inductions.The most general reaction is by PSMA711(sequence numbering: 7) induction, in 29 patients that analyze, its induction is the reactive polypeptide of 25 wherein.

embodiment 3

Its reactivity of specificity HLAII class binding peptide being included in mixtures of polypeptides detects after the treatment of embodiment 1.

Synthetic peptide and stimulation

For stimulating synthetic peptide with functional test, be HIV source property epi-position (HIV gag164-181:YVDRFYKTLRAEQASQEV(sequence numbering: 15), negative control); 13) and survivin 97-111:TLGEFLKLDRERAKN(sequence numbering PSMA459-473:NYTLRVDCTPLMYSL(sequence numbering:: 14).

Specific T-cells amplification in vitro

Different time points is during inoculation collected the peripheral blood lymphocytes of 15 and No. 26 patients with prostate cancer and with 90% hyclone and 10%DMSO cryopreservation in liquid nitrogen.After thawing, approximately 5 * 10 ⁶individual cell culture (24 porocyte culture plates, Greiner Bio-One company, Frickenhausen, Germany) in the IMDM culture medium, 50 μ g/ml streptomycins (all Biowhittaker company products that are that supplement 50U/ml penicillin, Verviers, Belgium), the beta-mercaptoethanol under 10% hot deactivation human serum (c.c.pro, Neustadt, Germany) and 50 μ M37 ° C condition and 7.5% CO ₂.The synthetic HLA II class binding molecule that adds storage on the 1st day, the concentration of every part is 5 μ g/ml, and in culture fluid, supplements recombinant human il-2 (r-hIL2, R & D Systems company in the 3rd, 5,7 and 9 days of T cytositimulation, Wiesbaden, Germany).After stimulating 12 days, cell harvesting, cleaning, counting and stimulate again 6 hours with peptide (10 μ g/ml).According to MACS IFN-γ secretion algoscopy (German Bel's Ji is executed Gladbach Mei Tian Ni biotech company), with IFN-γ, catch reagent and IFN-γ PE antibody labeling IFN-γ secretory cell, then use FACSAria(BD life sciences company) sorting on 96 orifice plates of the external source feeder cells (PBMC+LG2-EBV) that contain IMDM10%HS, 150U/ml IL-2,1 μ g/ml PHA and irradiation.Within every 4 days, add IL-2(150U/ml), feeder cells are within every 3 weeks, to add IL-2.

The dyeing of the cell within a cell factor

According to operation instructions, there is the BD life sciences company of Golgi-STOP(Heidelberg) and brefeldin A (10 μ g/ml, Sigma Ao Ruiqi company) time, in standard detection, collect, clean and use HIV, PSMA and survivin peptide or ionomycin (being respectively 50ng/ml and 1 μ M) the stimulating effect thing of 5 μ g/ml.Hatch after 4-6 hour cell PBS2%FCS0.02%NaN ₃clean and use monoclonal antibody (MoAb) CD4-APC-Cy7(BD life sciences company) and CD8-PE-Cy7(Beckman Coulter Inc.) in 4 ° of C dark, dye 20 minutes.After cleaning step, with Cytofix/Cytoperm reagent (BD life sciences company), cell can be permeated, then use monoclonal IFN-γ – FITC antibody (BD life sciences company) to IFN-γ dyeing in born of the same parents.Use software Diva on cell instrument CantoII, to carry out cell collection and analyze (BD life sciences company) with FlowJo.In Multifunctional inspection, CD4-APC-Cy7 and CD8-PerCP are for cell membrane dyeing, and IFN-g PE-Cy7, TNF-navy blue, IL-2-PE are for dyeing (from BD life sciences company, except TNF-navy blue is from Biolegend company) in born of the same parents.Between stimulation period, each test adds the CD107a-FITC(BD life sciences company of 1.5 μ l).

Use the function of autologous dendritic cell check T cell clone

By cultivate mononuclear cell 7 days in being supplemented with IMDM10%HS, the 1%PenStrep of 1000U/ml IL-4 and 800U/ml GM-CSF, 50 μ M beta-mercaptoethanols, produce the autologous DC of immaturity of cells of monocytic origin.In functional experiment, before intracellular cytokine dyeing, DC load be correlated with HLAII class binding peptide (HIC, survivin or PSMA, 10 μ g/ml) or with recombiant protein (survivin, PSMA or RAP-80,20 μ g/ml) impulse, be collected, clean several times and hatch 12 hours according to the ratio of DC and effector 1:5 with specific C D4+T cell clone.

In control experiment, when there being 1mM CaCl ₂time, with the 100 μ l PBS of pH7.2 and 10 μ g E.C. 3.4.21.64s (Dylan, Germany Macherey-Nagel company) 37 ° of C pretreatment, 1 μ g recombiant protein 2 hours.Or DC finally fixes 15 seconds in 0.05% glutaraldehyde (Sigma company).With 0.2M 1B (Sigma company) stopped reaction, DC is cleaned twice before load peptide or protein.

HLA II class restriction is measured and is used No. 15 and No. 26 pre-sensitization of patient's peptide PBMC action effect cells of 12 days and peptide load (10 μ g/ml peptides, in IMDM2%FCS, spending the night) EBV transformation cell lines is as stimulus object (being shown in HLA-DR, DP and DQ allele in Figure 21), its ratio is 1 effector: 1 peptide presenting cells, and measure peptide and offer.After jointly hatching 5 hours, the intracellular cytokine that completes as described above dyes.

Claims

1. a compositions, comprises at least two kinds of HLA binding peptides, wherein:

(a) at least two kinds of HLA binding peptides, have at least one peptide to contain sequence numbering: 23 epi-position or sequence numbering: 23 fusion rotein, 80 aminoacid that fusion rotein contains the relevant invariant chain N-end of HLA-DR antigen,

With

(b) at least two kinds of HLA binding peptides, there is at least one peptide to contain the epi-position of selecting from following one group of sequence numbering: sequence numbering: 1 to 11,13 to 22,24 to 42.

2. compositions according to claim 1, wherein, described compositions comprises at least two kinds by b) peptide that forms of group aminoacid sequence.

3. compositions according to claim 1, wherein said compositions comprises at least two kinds of peptides, be preferably at least 4 kinds of peptides, 10 kinds of peptides more preferably, by sequence numbering: 23 aminoacid sequence forms, a kind of peptide is to select from following one group of sequence numbering: sequence numbering: 1,2,5,7,9 to 11,13,14, and at least one peptide is to select from following one group of sequence numbering: sequence numbering: 15 to 22,24 to 42.

4. compositions according to claim 3, wherein, extra peptide is selected according to experimenter HLA group to be treated.

5. compositions according to claim 1, wherein, described compositions comprises at least 4 kinds by sequence numbering: the peptide that 23 aminoacid sequence forms, a kind of peptide is to select from following one group of sequence numbering: sequence numbering: 1,2,5,7,9 to 11,13,14, and at least one peptide is to select from following one group of sequence numbering: sequence numbering: 15 to 22,24 to 42.

6. compositions according to claim 5, wherein, at least one peptide is II class peptide.

7. according to the compositions described in arbitrary claim in claim 1 to 6, further comprise the mixture of immunological adjuvant or two or three kind of immunological adjuvant, such as GMCSF and imiquimod.

8. compositions according to claim 7, wherein, described immunological adjuvant contains Toll sample receptor stimulating agent, as Toll sample receptor-7 agonist.

9. according to the compositions described in arbitrary claim in claim 1 to 6, contain at least one antigen presenting cell, dendritic cell for example, as the autologous dendritic cell of peptide impulse or load peptide.

10. the compositions that is used for the treatment of carcinoma of prostate described in arbitrary claim in claim 1 to 9.

11. compositionss according to claim 10, wherein, described carcinoma of prostate be androgen responsive type and patient do not accept androgen-deprivation treatment.

12. compositionss according to claim 10, wherein, described carcinoma of prostate is androgen insensitivity type.

The method of the treatment carcinoma of prostate in 13. claim 1 to 9 described in arbitrary claim, comprises the compositions that gives patient's effective dose.

14. methods according to claim 13, wherein, described carcinoma of prostate be androgen responsive type and patient do not accept androgen-deprivation treatment.

15. methods according to claim 14, wherein, described carcinoma of prostate is androgen insensitivity type.