CN1311688A - Synthetic somatostatin immunogen for growth promotion in farm animals - Google Patents

Abstract

The invention provides peptides comprising somatostatin, or a sequence homologous to somatostatin, which is covalently linked to a helper T cell epitope and optionally to other immunostimulatory sequences. The present invention provides for the use of such peptides as immunogens to elicit the production in mammals of high titer polyclonal antibodies, which are specific to somatostatin. The peptides are expected to be useful in lowering somatostatin levels in mammals, thereby increasing growth rate and food utilization efficiency.

Description

Be used to promote the synthetic somatostatin immunogen of animal farm growth of animal

Invention field

The present invention relates to peptide combinations as the immunogen that promotes the feed lot growth of animal.The immunogen peptide of the present composition contains helper T cell epitope (Th), and it contains multiple II class MHC in conjunction with primitive, and has somatostatin (somatostatin) at any end of its C-or N-end.This peptide randomly, contains the invasin domain of the general immunologic stimulant of a kind of conduct.Helper T cell epitope and invasin domain are achieved the immunne response of anti-somatostatin self peptide.

Background of invention

Recently, about the understanding of nerve-endocrine and the growth of hormone factor affecting, and the fast development of biotechnology, for the growth rate of improving multiple feed lot animal varieties provides possible approach.The method of improving growth of animal by the regulating action of immunity neutralization change growth hormone is that application antibiotic, anabolic steroid and growth hormone a kind of valuable of existing method as growth promoter substituted.The application of this antibiotic in animal feeding it is believed that it is the reason of screening antibiotic resistance in some pathogen species, and it is producing unfavorable result (Witte, science, 1998 aspect control human infectious disease; 279:996).Use anabolic steroid and can promote the feed lot growth of animal, but in consumer, be out of favour.In Europe, the application of steroid not only all is restricted (Buttery andDawson, natural science progress, 1990 legally but also on public's suggestion; 49:459).Recombinant DNA technology can produce a large amount of metabolic hormones of directly animal being used.This application can promote growth.Yet if growth hormone is used as the method for improving the growth of animal situation, it only is a step need adopting.

Main growth hormone has: growth hormone (somatotropin) (growth hormone (growthhormone), GH), somatomedin-C (type-1 insulin like growth factor, IGF-1), somatocrinin (the GH releasing factor, GRF) and somatostatin (somatostatin) (GH release inhibiting factor).Growth hormone main may be used and be growth and lactogenic, yet, the hormone control of these phenomenons depend on complexity between some different hormones interact (Spencer, Livest Prod Sci, 1985,12:31).Therefore, the simple application of single hormone may be not enough to strengthen the productivity.

GH may bring into play two kinds of different effects.It is performance good effect aspect the increase of stimulated muscle protein synthesis (even be not all, also being that great majority are by the IGF-1 mediation), simultaneously because its lipolytic ability has potent catabolism effect.Comprehensive function is to increase carcass lean meat, reduces carcass fat, promotes growth usually, says at large and has improved efficiency of food conversion (Buttery and Dawson, natural science progress, 1990; 49:459; And, Spencer, Reprod Nutr Develop, 1987; 27 (2B): 581).

Use GH and promote that the effect application in practice of growth is insecure; May ignore may influencing that blood plasma level raises and causes among the receptor group because the administration of exogenous hormone has been ignored the sensitive interaction between various hormones.Effective application need of GH is frequently injected, so that the effective serum-concentration of continuous physiology of hormone is provided.It has increased the chaotic and dysgenic risk of generation of these equilibrium phase mutual relations.

Therefore, though can be by the purification goods of recombinant DNA technology mass production growth hormone, this technology can not provide ideal growth effect required transmission regulatory mechanism.And the application of recombinant human growth hormone in carnivore used in milch cow recently, caused consumers in general's opposition and dystaxia.Use growth hormone and promote ruminant and the accumulation of other feed lot animal meat, in Economic Commission for Europe, do not coordinated to pass through.Its coordination situation and political situation are similar to uses IGF-1 or the growth of GRF promotion feed lot animal varieties.

As a kind of replacement method that increases the growth stimulating hormone level, its proof can be equally or is more effectively removed the endogenous growth inhibitory substance.The main inhibiting substances of total meat growth is a somatostatin.Somatostatin is a kind of 14 amino acid whose ring-like peptides, and its structure is guarded between species.It is synthetic with 92 amino acid whose preceding somatostatin molecular forms, six kinds of peptides having known the somatostatin that comprises somatostatin itself and a kind of 28 amino acid form derive from preceding somatostatin molecule (Reichlin, laboratory clinical medicine magazine (J Lab Clin Med), 1987; 109:320).Somatostatin suppresses the release of some gastrointestinal hormones, and the release that suppresses GH, insulin, thyroxin, influences animal thus and absorbs nutraceutical ability, and influence it and guide these nutrients to enter the ability of tissue growth subsequently.

In rat, use the somatostatin antagonist find to stimulate its growth (Spencer eta1., the science of life (Life Sci), 1985,37:27), but also there is the shortcoming that needs every day and inject GH, IGF-1 and GRF in this processing.A kind of selection of putting into practice is to use the immunity of immune response inducing somatostatin neutralization.Concerning consumer and coordination organization, use immune system and promote that growth fraction is directly used hormone or desogestrel may be more acceptant.Spencer etc. at first in sheep by vaccine to somatostatin carry out immunity neutralization (Livest prodSci, 1983,10:469).

In a preliminary study, use twin St.Kilda lamb, the growth rate that the active immunity of somatostatin is caused handling lamb for the contrast lamb 176% (Spencer etal., animal reproduction (Anim Prod), 1981,32:376).Research subsequently can not repeat this result, but growth rate generally improves 15-20%.The somatostatin molecule all is identical in all pasture animal varietiess, has shown that the active immunity of anti-somatostatin can stimulate sheep (Spencer et al., Livest Prod Sci, 1983, the 10:25 with commercial significance; Laarveld et al., Canada's animal science magazine (Can J Anim Sci), 1986,66:77), cattle (Lawrence et al., animal science magazine (J Anim Sci), 1986,63:(Suppl) 215), pig and chicken (Spencer et al., Dom Anim Endocr, 1986, the 3:55) growth of Si Yanging.Table 1 has shown the general introduction of the examples of many successful that shows the effective immunity of the anti-somatostatin of pasture animal.

Except stimulating growth speed with make feeding time reduce by 20% (Spencer, Reprod NutrDevelop, 1987; 27 (2B): 581), the active immunity of anti-somatostatin also has beneficial effect aspect the transformation efficiency of food.Except since the quicker growth of animal save the food, in fact in growth course, more effectively utilize their food (Spencer et al., Livest Prod Sci, 1983,10:469), to change (Fadlalla etal., the J Anim Sci of small part owing to intestinal motive force, 1985,61:234; Faichney et al., Canadian animal science magazine, 1985,64 (supplementary issues) 93).This is handled the carcass composition without any appreciable impact (Spencer etal., 1983, the same), but indicates, and when the animal that kills with weight, handling animal may be thinner.All experimental datas are combined, active immunity may be a kind of stronger, safe and effective promotion growth instrument (Spencer, Dom Anim Endocr, 1986,3:55).

Existing bibliographical information designs and has tested several immunogen forms of somatostatin.For example, somatostatin is combined with protein carrier, render a service with enhance immunity.Yet protein carrier is too expensive for the Economic Application of pasture animal.And effective immune dependent of somatostatin is in the binding site of somatostatin and carrier.Even be not all also is to pass through the crosslinked preparation of glutaraldehyde coupling between the lysine residue on most of somatostatin protein carrier conjugate application somatostatin and the carrier protein.Therefore be used for link coupled two lysines on the somatostatin and be present in 12-aggressiveness function ring, when this conjugate is used as vaccine, may cause the remarkable forfeiture of natural somatostatin structure and the reduction of somatostatin cross reactivity.

And, it is in-problem that protein is connected in somatostatin, because most of immunne response are pointed to carrier rather than somatostatin (quality of carrier molecule is far longer than the quality of somatostatin molecule), and the immunity of using the hapten-carrier conjugate causes the inductive immunosuppressant of carrier (Schutz et al. usually, Journal of Immunology, 1985,135:2319).Therefore, seeking a kind of domestic animal, cheap and can stimulate the immunostimulant of the early stage potent immunne response of anti-somatostatin that is applicable to always.This immunostimulant is replied and is avoided the inductive inhibition of carrier.

An immunogenic key factor that influences the somatostatin synthetic peptide based immunogens is that it is presented to immune through the t helper cell epi-position.In the past, this was to provide by the carrier protein of following above-mentioned shortcoming.Also with provide as hybridization polypeptide through the recombinant dna expression system (Riggs, US 4,812,554; US 4,563, and 424; And Xu et al., Chinese science (Sciencein China) (B volume), 1994; 37:1234).The synthetic peptide of T-helper epitopes (Th) that can also be by containing target hapten B cell site and suitable host is easier and provide more easily.This peptide except antibody combining site also with the reaction of helper T-cell receptor and class (Babbitt et al., nature, 1985,317:359), therefore stimulate the strict site-specific antibodie of target antibody binding site (target site) replied.The complete synthetic peptide based immunogens of somatostatin will utilize the following advantage that is superior to carrier conjugates and recombinant polypeptide: this product defines through chemistry, easy quality control, it is very stable, do not need the downstream that takes a lot of trouble, do not need meticulous process units, the immunne response that produces is site-specific, thus replying of having avoided not expecting, and for example epi-position suppresses.

The immunogenic immunogenicity of synthetic somatostatin can followingly be optimized: (1) makes somatostatin combine with the Th site that mixes of selection, so that most of population is replied it; (2) somatostatin is combined with the Th of the increased functionality that forms through combinatorial chemistry, thereby provide variable immunne response for a kind of population; (3) limit the required conformational characteristic of stablizing somatostatin by ring-type.

Be called as the epi-position that mixes Th and induce effective T cell auxiliary, and can with reduced immunogenicity and can not provide potent immunogenic B cell epitope to combine very itself.Designing the good Th/B cell epitope chimeric peptide that mixes can induce Th to reply in most of members of the genetic diversity population of performance multiformity MHC haplotype and antibody response subsequently.Mix Th and can origin come from potent immunogenic particular sequence and provide, these potent immunogens comprise Measles virus F albumen and hepatitis B virus surface antigen.Some known Th of mixing have been presented at reinforcement, and more the peptide aspect of reduced immunogenicity is effective than decapeptide hormone LHRH (US 5,759,551).

The length scale scope of potent Th epi-position is about 15-30 amino acid residue, has common architectural feature usually, may contain the specific markers sequence.For example, common trait is to have the amphiphatic molecule spiral, and it is that hydrophobic amino acid residue occupies spiral one side, electrically charged and polar residues occupy on every side the α-Luo Xuanjiegou of face (Cease etc., NAS's scientific advance, 1987,84:4249-4253).The Th epi-position also contains other one-level amino acid pattern usually, and for example Gly or charged residue are followed two to three hydrophobic residues, follow an electrically charged or polar residues thereafter successively.This pattern is called as the Rothbard sequence.Find that also the Th epi-position follows 1,4,5,8 rules usually, one of them with the residue of positive electric charge after the 4th, the 5th and the 8th follow hydrophobic residue.Because all these structures are made up of common hydrophobic, electrically charged and polar amino acid, every kind of structure can in a Th epi-position, exist simultaneously (Partidos etc., hereditary Journal of Virology (J Gen Virol), 1991,72:1293-99).Even be not all,, great majority contain at least a above-mentioned periodicity but mixing t cell epitope.These features can be inserted the design in idealized artificial T h site, comprise the Th epi-position of combination.Design about combination Th site provides MHC-in conjunction with the variable position of primitive and the tabulation of preferred amino acid (Cease et al., NAS's scientific advance (Proc Natl Acad SciUSA), 1987; 84:4249-4253); And the method (Wang etc., WO 95/11998) of the library peptide of the synthetic antigen library that is called as structure of preparation combination Th or SSAL is disclosed.Therefore, can be with 1,4,5,8 rules with making up MHC-is applied to the variable of SSAL and degeneracy site in conjunction with primitive positional alignment, and the residue of selecting these sites, so that farthest increase is to the immunne response scope (WO 95/11998) of artificial T h.

Peptide based immunogens is more pliable and tougher than protein usually, is difficult for keeping any preferred structure.Therefore be useful by introducing ring-type restriction stabilized peptide immunogen.The conformation of target epi-position can be simulated and keep to the peptide based immunogens of correct cyclisation, therefore induce the antibody (Moore that those sites on the real molecule is had cross reactivity,＜synthetic peptide---users' guidebook (Synthetic peptide s A User ' sguide)〉the 2nd chapter, Grant compiles, WH Freeman and Company:New York, 1992, pp 63-67).

With the peptide based immunogens of peptide technology previously discussed and the design of peptide design element, the design of promptly miscellaneous potent Th epi-position, Th SSAL combined peptide and ring-type restriction, be the effectively synthetic immunogenic basis of somatostatin.This peptide is for presenting somatostatin by optimizing location and cyclisation, and extensive reactive Th to reply be preferred.Therefore, have been found that and contain independent Th epi-position or for example that the functional site in a kind of invasin domain (US5759551) and the 12 aggressiveness circuluses does not have peptide that the ad hoc structure of the somatostatin (as target site) of disturbed complete form arranges, and to produce aspect the anti-somatostatin antibody be effective stimulating in conjunction with a kind of general immunostimulant.

Summary of the invention

The present invention relates to a kind of immunogenicity peptide combinations that contains synthetic peptide, it can induce the antibody of anti-somatostatin, and it causes the inhibition of the plain inhibin level of pasture promoting animal growth, promotes growth and improves efficiency of food conversion.Specifically, peptide of the present invention has a kind of carboxyl or aminoterminal Th epi-position that is connected in somatostatin (SEQ ID NO:1) or somatostatin peptides analog.Randomly, this peptide has a kind of invasin domain (SEQ ID NO:2) as general immunologic stimulant.These peptides can be effective as immunogen, can increase serum growth hormone level in immune animal, increase thereby improve animal farm animal daily weight.

Another aspect of the present invention provides a kind of antigen composition, its peptide combinations of the present invention and one or more medicines that contain a kind of immune effective dose can be accepted bacterin preparation, and the dosage explanation, to such an extent as to generation is at the immunization therapy antibody in target somatostatin site.This peptide combinations is useful for improving the animal farm growth of animal.

Further aspect of the present invention relates under the condition by the functional antibodies of inducing anti-described somatostatin at certain hour and being enough to gives one or more peptides of the present invention of administration, to increase the method for level of growth hormone in the mammal blood circulation.

Of the present invention relating in one aspect to again a kind ofly has about 30 to the synthetic peptide of about 90 amino acid whose immunity, it contains the peptide analogues of a kind of helper T cell (Th) epi-position, somatostatin (SEQ IDNO:1) or somatostatin, the spacer of separating the immunogen domain and optional general immunostimulation site, for example invasin domain (SEQ ID NO:2).This of this peptide three kinds of immunogenic structure territories element and spacer can be covalently bound with any order, as long as the haptenic immunoreactivity of this peptide keeps the immunoreation that maybe can produce with the somatostatin self peptide basically.Detailed Description Of The Invention

The present invention relates to a kind of new peptide combinations, be used for producing somatostatin is had the specific height polyclonal antibody of tiring.The height locus specificity of this peptide combinations has reduced the production of antibodies at uncorrelated site on the carrier protein.Therefore, the invention further relates to a kind of method that in the animal farm animal, promotes growth.

Somatostatin (SEQ ID NO:1) is a kind of becate shape peptide hormone, itself right and wrong-immunogenic, all the more so as an autoantigen.Can by with this small peptide chemical coupling to a kind of carrier protein, for example, keyhole limpet hemocyanin (KLH) or by recombinant dna expression with itself and a kind of carrier polypeptide, for example, hepatitis B surface antigen merges, and carries out immune strengthening.The major defect that is somebody's turn to do " somatostatin-carrier " vaccine is that the overwhelming majority of the antibody that combination forms is the not function type antibody at carrier protein or polypeptide, and the possibility that exists epi-position to suppress.Immunogen of the present invention is synthetic fully peptide, and it has reduced irrelevant production of antibodies, thereby induces more the immunne response at somatostatin.Yet, because somatostatin be a kind of non--immunogenicity T cell-dependence antigen, its immunogenicity exogenous Th epi-position that places one's entire reliance upon.The of the present invention peptide covalently bound with mixing the Th epi-position provides this feature.Immunogen of the present invention all is the locus specificity immunoreation, so that provide effective growth for domestic animal.

The invention provides specific embodiment as peptide embodiment of the present invention.These embodiment provide synthetic immunostimulation element to be connected with somatostatin peptides, thereby produce potent somatostatin-reactive antibody in the diversified host group of heredity.These antibody then cause the inhibition of somatostatin function, promote the growth of domestic animal thus effectively.

For active immunity, the term that the present invention refers to " immunogen " relates to a kind of peptide combinations, and the antibody that it can induce anti-somatostatin causes the inhibition of somatostatin level in the mammal.Peptide combinations of the present invention comprises and contains the peptide that mixes helper T cell epitope (Th epi-position).This peptide (for example, Gly-Gly) is covalently attached to somatostatin peptides, thereby makes its N-that is connected in the target somatostatin peptides or C-end, so that induce effective antibody response by a kind of spacer.Immunogen can also comprise a kind of general immunostimulation element, and is for example, a kind of from Yersinia genus (Brett etc., European Journal of Immunology, 1993,23:1608-1614) the invasin domain of (SEQ ID NO:2).This invasin domain combines with the Th peptide by a kind of spacer.

Peptide of the present invention can be expressed from the next:

H ₂N-(A) _n-(somatostatin peptides)-(B) _o-(Th) _m-X

Or

H ₂N-(A) _n-(Th) _m-(B) _o-(somatostatin)-X

Wherein:

H ₂N is the terminal α-NH of the N-of peptide conjugates ₂,

Each A is a seed amino acid independently, or a kind of general immunostimulatory peptides;

Each B be selected from aminoacid ,-NHCH (X) CH ₂SCH ₂CO-,-NHCH (X) CH ₂SCH ₂CO (Lys-of ε-N) ,-NHCH (X) CH ₂The S-succinimido (Lys-of ε-N) and-NHCH (X) CH ₂S-(succinimido)-;

Each Th is a kind of aminoacid sequence that contains helper T cell epitope independently, or its immunostimulant analog or fragment;

Somatostatin peptides is somatostatin or its cross reactivity and immunologic function analog;

X is a kind of amino acid whose α-COOH or α-CONH ₂

N is from 1 to about 10;

M is from 1 to about 4; With

O is from 0 to about 10.

Peptide based immunogens of the present invention comprises about 20 to 100 amino acid residues, and preferred about 25 to about 80 amino acid residues, more preferably from about 25 to 60 amino acid residues.

If A is a seed amino acid or a kind of general immunostimulation element, for example, Inv, it can be covalently attached to the N-end (shown in general formula) of peptide based immunogens or be connected to the terminal (not shown) of C-.

If A is a seed amino acid, it can be any natural or non-natural aminoacid.Non-natural aminoacid includes but not limited to D-aminoacid, Beta-alanine, ornithine, nor-leucine, norvaline, hydroxyproline, desiodothyroxine, γ-An Jidingsuan, homoserine, citrulline etc.Natural amino acid comprises alanine, arginine, agedoite, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine and valine.And if m is an aminoacid greater than 1, two or three A groups, each aminoacid can be identical or different independently so.

If A is a kind of domain of invasin, then it can be from the proteic immunostimulation epi-position of Yersinia kind invasin.This immunostimulatory properties derives from this invasin domain and T cell, the ability of the beta 1 integrin interaction of molecules that exists on particularly activatory immunity or the memory T cell.Brett etc. have described particular sequence (European Journal of Immunology, 1993 with the interactional invasin domain of beta 1 integrin; 23:1608).Described a kind of preferred implementation that is used to connect the invasin domain (Inv) that mixes the Th epi-position in US 5759551, it inserts this paper through reference.The Inv domain preferably has following sequence:

Thr-Ala-Lys-Ser-Lys-Lys-Phe-Pro-Ser-Tyr-Thr-Ala-Thr-Tyr-Gln-Phe

(SEQ ID NO:2) or for deriving from its immunostimulation analog of another kind of Yersinia kind invasin albumen respective regions.Therefore this analog can contain replacement, disappearance or the insertion of amino acid residue, with the variation of adapted strain system, as long as analog keeps immunostimulatory properties.

In one embodiment, n is 1, and A is α-NH ₂In another embodiment, n is 4, and A is α-NH ₂, invasin domain (Inv), glycine are connected successively with glycine.

B is a kind of spacer, is a kind of above-mentioned natural amino acid or alpha-non-natural amino acid.Each B is identical or different independently.The aminoacid of B can also provide a kind of spacer mixing between Th epi-position and somatostatin peptides (for example, SEQ ID NOS:1) or its cross reactivity and the immunologic function analog, for example Gly-Gly.Except making Th epi-position and B cell epitope, be outside somatostatin peptides and the immunogenic analog physical separation, the Gly-Gly spacer can also destroy the Th epi-position and combine all artificial secondary structures that produce with somatostatin peptides and its cross reactivity and immunologic function analog, has therefore weakened the interference between Th and/or the B cell response.The aminoacid of B can also constitute a kind of spacer as the elasticity hinge that strengthens Th and IgE domain compartmentation.Found the example of the sequence of coding elasticity hinge at the heavy chain immunoglobulin hinge region.The common proline rich of pliability hinge sequence.A kind of useful especially pliability hinge is sequence Pro-Pro-Xaa-Pro-Xaa-Pro (SEQID NO:3), and wherein Xaa is an arbitrary amino acid, is preferably aspartic acid.The conformation that is provided by the aminoacid of B makes the interaction between peptide based immunogens of the present invention and suitable Th cell and the B cell more effective at interval, has strengthened the immunne response to Th epi-position and antibody-inductivity epi-position and their cross-interaction and immunologic function analog thus.

Th is a kind of aminoacid sequence (natural or alpha-non-natural amino acid) of the Th of containing epi-position.The Th epi-position can be made of continuous or discontinuous epi-position.Therefore each aminoacid that is not Th must be the part of epi-position.Therefore, the Th epi-position comprises the analog and the fragment of Th epi-position can strengthening or stimulate the immunne response to somatostatin peptides and immunologic function analog thereof.Show as immune structure territory and miscellaneous Th epi-position and in animal and human group, have height and reactive widely (Partidos etc., 1991 with extensive difference MHC type; US5759551).The Th domain of peptide of the present invention has 10 to 50 aminoacid approximately, preferably has 10 to 30 aminoacid approximately.When a plurality of Th epi-positions (being m 〉=2) occurring, then each Th epi-position is identical or different independently.The Th fragment is the continuous part of Th epi-position, and it is enough to strengthen or stimulates immunne response to somatostatin peptides (SEQ ID NO:1) and immunologic function analog thereof.

Epi-position of the present invention comprises the Th that derives from foreign pathogens, include but not limited to, for example hepatitis B surface antigen and cAg helper T cell epitope (HBs Th and HBc Th), pertussis toxin, PT helper T cell epitope (PT Th), tetanus toxin helper T cell epitope (TTTh), Measles virus F albumen helper T cell epitope (MV _FTh), chlamydia trachomatis major outer membrane albumen helper T cell epitope (CT Th), diphtheria toxin, diphtherotoxin helper T cell epitope (DT Th), plasmodium falciparum ring sporinite helper T cell epitope (PF Th), schistosoma mansoni trisaccharide phosphate isomerase helper T cell epitope (SM Th), and escherichia coli TraT helper T cell epitope (TraT Th).The present invention selects to list as the SEQID NOS:2-9 and the 42-52 of the deutero-Th of pathogen in US 5759551 that mixes the representative example of Th; CT Th P11 is at Stagg etc., immunology, and 1993, list among the 79:1-9; HBc peptide 50-69 is at Ferrari etc., Journal of Clinical Investigation, and 1991, to list among the 88:214-222, they are through with reference to inserting this paper.In addition, the Th epi-position comprises idealized artificial T h (for example, SEQ ID NOS:14) and artificial SSAL Th (for example, SEQ ID NOS:7,30,31).The peptide that comprises SSAL Th is synthetic simultaneously in order with somatostatin and other sequence in a solid-phase peptide is synthetic.The Th site also comprises the functional immunity analog.Immunologic function Th analog comprises the fragment of immunostimulant analog, cross reactivity analog and any of these Th epi-position.Functional Th analog further is included in conservative replacement in the Th epi-position, additional, deletion and inserts one to 10 amino acid residue, and does not change the Th stimulatory function of Th epi-position basically.

Of the present invention synthetic peptide by the following formula description:

(A) _n-(Th) _m-(B) _o-(somatostatin peptides)

Or

(A) _n-(somatostatin peptides)-(B) _o-(Th) _mThe N end or the C that have by spacer B and somatostatin peptides and cross reactivity and functional immunity analog hold covalently bound Th epi-position.

According to the present invention, somatostatin peptides (for example, SEQ ID NO:1) cross reactivity and functional immunity analog may further include conservative replace, additional, deletion or insert to about four amino acid residues, as long as this peptide analogues can be induced the immunne response with the somatostatin peptides cross reaction.Conservative replace, additional and insert and can realize with the natural or alpha-non-natural amino acid of the present invention's definition.

Preferred peptide based immunogens of the present invention is to contain somatostatin peptides or its cross reactivity and functional immunity analog; Spacer (for example, Gly-Gly); The TH epi-position is HBs Th (SEQID NO:1 5), HBc Th (SEQ ID NO:4), MVF Th (SEQ ID NOS:21,29), PT Th (SEQ ID NO:6), TT Th (SEQ ID NO:5), CT Th (SEQID NO:27), DTTh (SEQ ID NO:28), artificial T h (for example, SEQ ID NOS:7,14,30,31) or its analog; And randomly, the peptide of a kind of Inv domain (SEQ ID NO:2) or its analog.

The peptide combinations that contains the mixture of peptide based immunogens of the present invention and one or more Th epi-positions can be renderd a service in enhance immunity in the population widely, and produces the improved immunne response to somatostatin peptides thus.

Peptide based immunogens of the present invention can be by chemical synthesis preparation known to a person of ordinary skill in the art.Referring to, for example, Fields etc., " synthetic peptide: users' guidebook " chapter 3, Grant, W.H. compiles, Freeman and Co., NewYork, NY, 1992, p.77.Therefore; can use solid-phase synthesis---mate the Merrifield technology automatically; with t-Boc or Fmoc chemical compound protection α-NH2; and the aminoacid of application side chain protected; for example, synthesize peptide on 430A or the 431 type applying biological system peptide synthesizers (the Applied Biosystems peptide Synthesizer Models430A or 431).Can be by being provided for being attached to a locational selection ispol of particular variable, preparation contains the peptidic constructs of the SSAL of Th epi-position.

After finishing the assembling of required peptide based immunogens,, excising peptide, and make the functional group on the amino acid side chain take off sealing from resin according to the standardization program process resin.By HPLC purification free peptide, identify biochemical character by for example amino acid analysis or by order-checking.The purification of peptide and characterized method are known for those skilled in the art.

Immunogen of the present invention can also be polymeric.Can pass through, for example utilize conventional method make glutaraldehyde and lysine residue-NH ₂Between the realization that reacts.By another kind of method, promptly by an additional cysteine being added to synthetic " A-Th-spacer-(somatostatin peptides) " or " (growth swash inhibin peptide)-spacer-(Th) _m-(A) _n" immunogenic N-end, make synthetic " A-Th-spacer-(somatostatin peptides) " or " (growth swash inhibin peptide)-spacer-(Th) _m-A " immunogen polymerization or combined polymerization.Can also be by directly synthetic required peptide structure on a kind of ramose poly lysyl core resin, the immunogen of the present invention (Wang etc., science, 1991 that prepare a kind of branched polymer; 254:285-288).

Perhaps, long synthetic peptide based immunogens can be synthetic by known recombinant DNA technology.The manual of standards of any relative dna technology provides the detailed method for preparing peptide of the present invention.In order to make up the gene of code book invention peptide, translate into a kind of nucleotide sequence with aminoacid sequence is reverse, the preferred utilization is applied to the organic optimizing codon that gene will be expressed therein.Then, prepare synthetic gene, pass through the overlapping oligonucleotide of composite coding peptide and some necessary controlling elements usually.Synthetic gene is inserted suitable cloning vehicle, obtain recombinant and its evaluation.Expression of peptides under the felicity condition that the expression system of selecting and host are fit to then.Utilize standard method purified peptide and identification mark.

The effect of peptide combinations of the present invention can be by for example containing peptide of the present invention, SEQ IDNOS:8-13,16-20,22 immunogenic composition injection animal, rat for example, monitor then the hormone immunity of somatostatin and cross reactivity and functional immunity analog is replied, confirm, as the detailed description among the embodiment.

Another aspect of the present invention provides a kind of pharmaceutical composition that contains a kind of one or more peptide based immunogens of the present invention of immune effective dose in the acceptable transfer system of a kind of medicine.Therefore, can use other conventional ingredient that provides in adjuvant, drug acceptable carrier or other vaccine combination peptide of the present invention is mixed with a kind of peptide combinations.Adjuvant of the present invention or emulsifying agent be can be used for and Alumen, incomplete Freund's adjuvant, liposyn, Saponin, Squalene, L121, emulsigen, single phosphatidyl lipid A (MPL), dimethyl-two-octadecyl bromination ammonia (DDA), QS21, ISA51, ISA35, ISA206 and ISA 720 comprised, and other effective adjuvant and emulsifying agent.Described preparation comprises rapid release and/or slow releasing preparation, and the preparation of inducible system immunity, and it can pass through, and for example, seals immunogen or immunogen and microgranule co-administered are realized with microgranule.Those of ordinary skills can easily determine described preparation.Immunogen of the present invention can be by any conventional route administration, comprises subcutaneous, oral, intramuscular or other non-intestinal or intestinal administration.Equally, immunogen can be used as single dose or multiple dose administration.Those of ordinary skills can easily determine immunization protocol.

Peptide combinations of the present invention contains a kind of one or more peptide based immunogens of the present invention and a kind of medicine acceptable carrier of effective dose.Compositions in a kind of suitable dosage unit forms contains the immunogen of the about 0.5 μ g of per kilogram of body weight to about 1 mg usually.If with multiple dose administration, can be divided into every dosage unit preparations appropriate amount easily.For example, predose, for example 0.2-2.5 mg; Preferred 1 mg will use repeated doses (enhancing amount) by the preferred intramuscular injection administration of injection then by the immunogen that peptide combinations of the present invention is represented.Dosage will be decided according to patient's age, body weight and general health situation, as vaccine and treatment field are known.

Can be to the synthetic immunogenic immunne response of somatostatin peptides by being captured in (vaccine, 1991 such as O ' Hagan; 9:768) in the biodegradable microgranule of Miao Shuing or send improvement on it.Can with immunogen with or be not encapsulated into the biodegradable microgranule with adjuvant, reply with reinforced immunological, comprise the local mucous membrane immunity, and be provided for oral administration and be used to possess the time-delay of administration or the time sustained release of period answer (O ' Hagan etc., 1991; With Eldridge etc., 1991; 28:287-294).

Concrete peptide based immunogens and compositions are provided in the following embodiments, have been used to illustrate the present invention.The purpose of these embodiment only is explanation, scope of the present invention is not constituted the restriction of any way.

Embodiment 1

The conventional method of synthetic somatostatin peptides construct

By the Merrifield solid phase synthesis technique, mate on the peptide synthesizer (430,431 and 433A type) peptide of listing with the synthetic respectively table 2 and 3 of Fmoc chemical compound automatically in the applying biological system.Can be by being provided for the amino acid needed mixture on the indicated ad-hoc location in chemical coupling to a design, preparation contains the synthetic antigen library (SSAL) of structure, for example is called as the peptidic constructs in the artificial T h site of " 1,4,9PALINDROMIC " (SEQ ID NO:7).After finishing the assembling of required peptide, use trifluoroacetic acid, excising peptide, and make the blocking group on the amino acid side chain take off sealing from resin according to the standardization program process resin.For cyclic peptide, the peptide that excises was dissolved 48 hours in the aqueous solution of 15%DMSO, to promote the formation of interchain disulfide bond between the cysteine.

Excision, the peptide that extracts and clean pass through the HPLC purification, by mass spectrum and reversed-phase HPLC identification mark.

Target antigen peptide shown in the peptide that is labeled as " b " in the peptide code hurdle the synthesizes Th site that is linked in sequence.The TH site of using comprises, for example, and from the HBs Th (SEQ IDNO:15) of hepatitis B virus be referred to as the new artificial T h site of " 1,4,9PALINDROMIC ".The peptide that is labeled as " c " is the variant with Inv domain immunostimulatory peptides (SEQ ID NO:2) order synthetic " b " construct.The peptide that is labeled as " d " is that (for example, the peptide of somatostatin-Th), be labeled as " e " is the reverse thing of " c " construct (for example a, somatostatin-Th-Inv) for the reverse thing of " b " construct.Synthetic " b ", " c ", " d " and " e " construct have the Gly-Gly spacer, separate target antigen site and Th site, separate Th and Inv immunostimulation site.

Embodiment 2

Estimate the immunogenic conventional method of somatostatin peptides

As experiment immunization scheme of listing below and the explanation of determining immunogenic serological analysis, in every group of 4 or 5 rats, estimate somatostatin peptides immunogen (for example, SEQ ID NOS:8-13, the 16-20,22 and 24 shown in table 2 and 3).Standard test design: immunogen: (1) individual peptides immunogen; Or

(2) contain the mixture that waits the mole peptide based immunogens that specifies just like each embodiment.Dosage: 100 μ g among each immune 0.5 mL, except as otherwise noted.Approach: intramuscular, adjuvant except as otherwise noted: (1) Freund's complete adjuvant (CFA)/Freund (IFA); Or

(2) 0.4% Alumen (aluminium hydroxide);

CFA/IFA group is accepted CFA in 0 week, subsequently several weeks accept IFA.All dosage of Alumen group are accepted same preparation.Dosage regimen: 0,2 and 4 weeks; 0,3 and 6 weeks, or explanation is arranged in addition.The blood sampling scheme: 0,3,6 and 8 weeks, or explanation is arranged in addition.Animal varieties: the every treated animal number of Sprague-Dawley rat: 4 or 5 rat/group analysis: analyze the anti--active specific ELISA of peptide of each immune serum, the solid phase substrate is cyclisation somatostatin peptides (SEQ ID NO:1).

Gather blood, be processed into serum, store, be used for tiring by ELISA mensuration with target antigen peptide.

On the 96 holes flat microdroplet flat board of the somatostatin peptides of using cyclisation in advance (SEQ ID NO:1), determine the activity of anti--somatostatin antibody by ELISA (Enzyme Linked Immunoadsorbent Assay) as the immunoadsorbent coating.The separatory (100 μ L) such as peptide based immunogens solution that with concentration are 5 μ g/ml were in 37 ℃ of insulations 1 hour.Add in the peptide coating hole with 3% gelatin/PBS solution, be incubated 1 hour once more, the sealing flat board in 37 ℃.The flat board of dry sealing is used for analyzing then.The separatory (100 μ L) such as grade of experiment immunization serum carries out ten times of serial dilutions since 1: 100 dilution factor in the diluted sample buffer, add then in the peptide spread plate.Dull and stereotyped in 37 ℃ of insulations 1 hour.

Use 0.05%PBS/TWEEN ^Washing plate 6 times.Be added in conjunction with the 100 μ L horseradish peroxidase labelled goat of suitably diluting in the dilution buffer liquid (phosphate buffer and the normal goats serum that contain 0.5M NaCl)-anti--species specificity antibody.With flat board in 37 ℃ of insulations 1 hour, then as above-mentioned flushing.Between adding then-the phenylenediamine substrate solution wait separatory (100 μ L).Color reaction carried out 5-15 minute, added 50 μ L 2N H then ₂SO ₄End the color reaction of enzyme.On plate reader, measure each hole at A _492nmContent.Trap is being carried out linear regression analysis, and with cut-off point A _492nmBe set on 0.5 the basis, calculate ELISA and tire.This cutoff is accurate, because this value of the dilution normal control sample of each analyzing and processing is lower than 0.15.

ELISA based on the Th peptide.ELISA based on the Th peptide carries out equally as the previously described somatostatin ELISA of the present invention basically, except the antigen application step, wherein microdroplet is dull and stereotyped was coated with 1 hour in 37 ℃ with the concentration of the specified individual Th peptide that derives from its corresponding somatostatin vaccine constructs (for example, SEQ IDNOS:14 and 15 peptide) with 5 μ g/ml.

Embodiment 3 combinations have the immunogenicity research of the somatostatin antigenic peptides of various immunostimulation elements

Somatostatin peptides immunogen shown in the table 2 is for example understood the peptide variant of the present invention that following formula is represented:

(A) _n-(Th) _m-(B) _o-(somatostatin peptides)

Or

(A) _n-(somatostatin peptides)-(B) _o-(Th) _mWherein

A is a seed amino acid, α-NH ₂, or Inv (SEQ ID NO:2);

When A is a seed amino acid or Inv, it can be connected to any end of N-end or C-end;

B is a glycine;

Th is a kind of helper T cell epitope that derives from foreign pathogens, for example, and HBc _50-69Th (SEQ ID NO:4), TT _615-631Th (SEQ ID NO:5), PT _147-176Th (SEQID NO:6) or and artificial T h, for example, 1,4,9 PALIDROMIC Th (SEQ IDNO:7);

N is 1, and m is 1, and o is 2.

These peptides are synthetic together with other peptide, and the immune serum of generation is used to carry out the immunogenicity evaluation.Somatostatin-the specific antibody of most of peptides inducement efficient valency in immune host of the Th epi-position that is integrated with various forms and orientation shown in the table 2.Comparatively speaking, the somatostatin peptides (p1348a, SEQ ID NO:1) that does not contain Th does not have immunogenicity.Yet it is preferred that some Th compares with other Th.For example, in table 2, have HBc Th (SEQ ID NO:4) p2134b (SEQ ID NO:8), have p2384b (SEQ ID NO:20), the artificial T h site of SynTh (1,2,3) (SEQ ID NO:14) and have PT _149-167The p2138b (SEQ ID NO:10) of Th (SEQ ID NO:6) has stronger immunogenicity than the p2135b with TT Th (SEQ ID NO:5) (SEQ ID NO:9), and all these all are preferable over almost at all do not have the immunogenic CTA8 of the having Th p2136b (SEQ ID NO:24) of (SEQ ID NO:23).In addition, for the immunogenicity of optimizing, the closure of Th site and somatostatin target site must be specific.The antibody response kinetics that compares p2253b (SEQ ID NO:11) and p2255d (SEQ ID NO:12).With 1,4, to place the C-end of somatostatin be preferred to 9PALINDROMIC Th (SEQ ID NO:7).From p1344b (SEQ ID NO:16) and p1349b (SEQ ID NO:22) relatively, MVF _258-277(SEQ ID NO:21's) is at the N-of somatostatin end than good position.Obviously the selection in each Th site and arrangement must be specific for preferred peptide of the present invention.

For the somatostatin peptides shown in the table 3, have been found that containing the somatostatin construct that mixes the Th epi-position has immunogenicity, and Inv combines with " Th " construct, they can improve the immunogenicity of somatostatin peptides.P1344b (SEQ ID NO:16) and the comparison of p1343c (SEQ ID NO:17) and comparison shows that of p1346b (SEQ ID NO:16) and p1345c (SEQID NO:19), add Inv domain (SEQ IDNO:2) the N-end of Th construct to, improved with the immunogenicity of replying the performance of the intensity of animal percentage rate, somatostatin-specific antibody titres and the persistent period of antibody response (＞35 week).Yet, p2134b (SEQ ID NO:8) and p2134c (SEQID NO:25) and with the immunogenicity comparative descriptions of p2134d (SEQ ID NO:26), after specific T h site combines, Inv may not possess immunostimulation, for Inv and HBcTh site (SEQ ID NO:4) combine on the N-end specific orientation than the orientation of C-end more preferably.For the example of p2135e (SEQ ID NO:13), Inv and TT _615-631Th (SEQ ID NO:5) produces a kind of effective immunogen in the combination of C-end.Therefore, the orientation of additional Inv and Th and Inv is specific for preferred peptide of the present invention.

For possessing the potent site immunogenic somatostatin construct that leads, at the immunostimulation element, for example, the Inv-MVF of the p1343c of table 3 (SEQ ID NO:17) _258-277The KKK-HBs of Th and p1346b (SEQ ID NO:18) _19-32Th-GG, antibody titer be＜1 Log ₁₀, antibody titer＞3 Log of the somatostatin of comparing ₁₀Therefore, the immunne response that is produced by synthetic peptide of the present invention is almost pointed to the somatostatin target site specially.

Embodiment 4

Other antigenic peptides of the present invention

The somatostatin peptides antigen example of having synthesized the peptide variant of the present invention that is expressed from the next, and produce immune serum.

(A) _n-(Th) _m-(B) _o-(somatostatin peptides)

Or

(A) _n-(somatostatin peptides)-(B) _o-(Th) _mWherein

Th is the helper T cell epitope that derives from foreign pathogens shown in a kind of table 4; Perhaps, a kind of helper T cell epitope that derives from the artificial T h epi-position shown in the table 5;

A is a seed amino acid, α-NH ₂, or Inv (SEQ ID NO:2);

When A is a seed amino acid or Inv, it can be connected to any end of N-end or C-end;

B is a glycine;

N is 1, and m is 1, and o is 2.

Table 1

Be used to promote the anti-somatostatin immunity of growing

Species	Growth percentage rate (%) compared with the control	List of references
			Sheep (twin)	????176	????Spencer?et?al.,1981
Sheep	????125	????Spencer?et?al.,1983
			Pig	????118	????Spencer?et?al.,1983
Cattle	????118	????Lawrence?et?al.,1986
			Chicken	????115	????Spencer?et?al.,?1986

With the vaccine of carrier protein-somatostatin conjugate as inoculation.

Table 2

Integrated the immunogenicity of the somatostatin antigenic peptides of various Th

The peptide code	The description of antigenic peptides	The aminoacid sequence of the auxiliary peptide of T	Adjuvant	Immunogenicity
							WPI	Reply (n=4)	Log 10ELISA tires b
				p1348a	Somatostatin a(Seq.IDNo.1)					0.4% Alumen (0,2,4WPI)	????6	????0	?????-
????8	????0	?????-
			p2134b				?HBc 50-69-GG-somatostatin (Seq.ID No.8)	SDFFPSVRDLLDTASALYRE (Seq.ID?No.4)	0.4% Alumen (0,2,4WPI)		????6	????100	????3.22
????8	????100	????3.18
				p2384b	SynTh (1,2,4)-GG-somatostatin (Seq.ID No.20)	KKKIITITRIITIITTID (Seq.ID?No.14)				0.4% Alumen	????8	????100	????3.12
p2135b	?TT 615-631-GG-somatostatin (Seq.ID No.9)	WVRDIIDDFTNESSQKT (Seq.ID?No.5)	0.4% Alumen (0,2,4WPI)				????6	????50	????2.528
				????8	????50	????2.518
							p2136b	?CTA8 106-130-GG-somatostatin (Seq.ID No.24)	ALNIWDRFDVFSTLGATSGYLKGNS (Seq.ID?No.23)	0.4% Alumen (0,2,4WPI)	????4	????0	????-
????6	????25	????2.437
			p2138b	?PT 149-176-GG-somatostatin (Seq.ID No.10)	KKLRRLLYMIYMSGLAVRVHVSKEEQYYDY (Seq.ID?No.6)	0.4% Alumen (0,2,4WPI)					????6	????50	????2.387
????8	????75	????2.349

????p2253b	1,4,9PALINDROMIC Th-GG-somatostatin (Seq.IDNo.11)	ISEIKGVIVHKIEGI ?MT?RT?TRM?TM ?L???????L??V(Seq.ID?No.7)	0.4% Alumen (0,2,4WPI)	????5	????25	????2.384
							????8	????100	????3.33
				????p2255d	Somatostatin-GG-1,4,9 PALINDROMIC Th (Seq.ID No.12)	ISEIKGVIVHKIEGI MT?RT????TRM?TM ?L?????????L??V				0.4% Alumen (0,3,6WPI)	????5	????75	????3.24
????8	????100	????3.12
			????p1344b				?MVF 258-277-GG-somatostatin (Seq.ID No.16)	GILESRGIKARITHVDTESY (Seq.ID?No.21)	CFA/IFA ????0,3,6		????5	????40	????2.745
????8	????80	????3.111
				????p1349	Somatostatin-GG-MVF 258-277(Seq.ID?No.22)	GILESRGIKARITHVDTESY (SEQID?NO:21)				CFA/IFA ????0,3,6	????5	????0	??????-
????8	????40	????2.782

^aThe sequence of somatostatin: AGCKNFFWKTFISC ^bThe experimental result of the serum that the reactive animal of ELISA-is collected.

Table 3

Immunogenic raising in conjunction with the somatostatin antigenic peptides of invasin domain

The peptide code	The description Seq.ID No. of antigenic peptides	The aminoacid sequence Seq.ID No. of the auxiliary peptide of T	Adjuvant	Immunogenicity
							WPI	The % response rate	Respondent's Log 10ELISA tires c
				?p1344b	?MVFTh 258-277-GG-somatostatin a(Seq.ID?No.16)	GILESRGIKARITHVD ?TESY (Seq.ID?No.21)				????CFA/IFA ????(0,3,6WPI)	????5	????40	????2.745
????8	????80	????3.111
			????10				????100	????2.917
????12	????80	????3.141
			????35				????40	????2.409
?p1343c	?Inv b-MVFTh 258-277-GG- (Seq.ID?No.17)	GILESRGIKARITHVD ?TESY (Seq.ID?No.21)							????CFA/IFA ????(0,3,6WPI)		????5	????40	????2.166
			????8	????80	????3.327
						????10	????100	????3.227
			????12	????100	????3.042
						????35	????100	????2.476
			?p1346b	?KKK-HBs 19-32Th-GG- (Seq.ID?No.18)	FFLLTRILTIPQSLD (Seq.ID?No.15)					????CFA/IFA ????(0,3,6WPI)	????5	????40	????3.690
????8	????60	????3.784
						????10	????60	????3.628
????12	????60	????3.580
						????35	????20	????3.879

Table 3 (continuous page or leaf)

????p1345c	?Inv-GG-KKK-HBs 19-32Th-GG (Seq.ID?No.19)	FFLLTRILTIPQSLD (Seq.ID?No.15)	????CFA/IFA ????(0,3,6WPI)	????5	????80	????3.755
							????8	????80	????4.141
				????10	????100	????3.428
							????12	????80	????3.770
				????35	????80	????3.166
							????p2134b	?HBc 50-69-GG-somatostatin (Seq.ID No.8)	SDFFPSVRDLLDTASA ?LYRE (Seq.ID?No.4)	0.4% Alumen 0,2,4, WPI	????4	????75	????3.00
????6	????100	????3.22
			????8	????100	????3.18
????p2134c	?Inv-GG-HBc 50-69-GG-somatostatin (Seq.ID No.25)	SDFFPSVRDLLDTASA ?LYRE (Seq.ID?No.4)				0.4% Alumen 0,2,4, WPI					????4	????75	????2.77
			????6	????100	????2.98
							????8	????100	????2.84
			????p2134d	Somatostatin-GG-HBc 50-69-GG-Inv (Seq.ID?No.26)	SDFFPSVRDLLDTASA ?LYRE (Scq.ID?No.4)					0.4% Alumen 0,2,4, WPI	????4	????0	?????-
????6	????0	?????-
						????8	????0	?????-
????p2135e	Somatostatin-GG-TT 615-631-GG-Inv (Seq.ID?No.13)	WVRDIIDDFTNESSQ ?KT (Seq?ID?No.5)							0.4% Alumen 0,2,4, WPI		????6	????100	????2.80
			????8	????100	????2.67

^aThe sequence of somatostatin: AGCKNFFWKTFTSC (Seq.ID No.1) ^bThe sequence of Inv (i.e. invasin land): TAKSKKFPSYTATYQF (Seq.ID No.2) ^cThe experimental result of the serum of collecting from the reactive animal of ELISA-.

Table 4

The aminoacid sequence of the Th epi-position in pathogen source

The description of Th	????SEQ?ID ????NO:	Aminoacid sequence
			?HBc 50-69?Th	????4	?SDFFPSVRDLLDTASALYRE
?TT 615-631?Th	????5	?WVRDIIDDFTNESSQKT
			?PT 149-176?Th	????6	?KKLRRLLYMIYMSGLAVRVHVSKEEQY YDY
?HBs 19-32?Th	????15	?FFLLTRILTIPQSLD
			?MVF 258-277?Th	????21	?GILESRGIKARITHVDTESY
?CTA8 106-130?Th	????23	?ALNIWDRFDVFSTLGATSGYLKGNS
			?CTP11?Th	????27	?TINKPKGYVGKE
?DT1?Th	????28	?DSETADNLEKTVAALSILPGHG
			?MVF 1?Th	????29	?LSEIKGVIVHRLEGV

Table 5

The aminoacid sequence of artificial T h epi-position

The description of Th	SEQ?ID?NO:	Aminoacid sequence
			Syn?Th(1,2,4)	????14	KKKIITITRIITIITTID
(1,4,9?PALINDROMIC)Th	????7	ISEIKGVIVHKIEGI MT????RT????TRM?TM L?????????????L??V
			(1,4,9 PALINDROMIC) simplifies Th	????30	ISEIKGVIVHKIEGI ?T??RT???TR??T
IS (1,4,9 PALINDROMIC) LF simplifies Th	????31	ISISEIKGVIVHKIEGILF ???T??RT???TR??T

Sequence table (1) general information:

(ⅰ) applicant: UNITED BIOMEDICAL INC., ET AL.

(ⅱ) denomination of invention: the synthetic somatostatin immunogen that is used to promote the animal farm growth of animal

(ⅲ) sequence number: 45

(ⅳ) address:

(A) address: Morgan﹠Finnegan

(B) street: 345 Park Avenue

(C) city: New York

(D) state: NY

(E) country: USA

(F) postcode: 10154-0054

(ⅴ) computer-reader form:

(A) medium type: floppy disk

(B) computer: IBM PC compatible

(C) operating system: PC-DOS/MS-DOS

(D) software: PatentIn Release#1.0, Version#1.25

(ⅵ) request for data formerly:

(A) application number: 09/100,415

(B) applying date: 20 JUNE 1998

(C) classification number:

(ⅶ) current request for data:

(A) application number: TBA

(B) applying date: 21-JUN-1999

(C) classification number:

(ⅷ) lawyer/agent's data:

(A) name: MARIA C.H.LIN

(B) number of registration: 29,323

(C) reference/file number: 1151-4155PCI

(ⅸ) telecommunication information:

(A) phone: 212-758-4800

(B) fax: the information of 212-751-6849 (2) SEQ ID NO:1:

(ⅰ) sequence signature:

(A) length: 14 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: the information of SEQ ID NO:1:Ala Gly Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr1 5 10Ser Cys (2) SEQ ID NO:2:

(ⅰ) sequence signature:

(A) length: 16 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:2:Thr Ala Lys Ser Lys Lys Phe Pro Ser Tyr Thr Ala1 5 10Thr Tyr Gln Phe

The information of 15 (2) SEQ ID NO:3:

(ⅰ) sequence signature:

(A) length: six amino acid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: the information of SEQ ID NO:3:Pro Pro Xaa Pro Xaa Pro1 5 (2) SEQ ID NO:4:

(ⅰ) sequence signature:

(A) length: 20 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:4:Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp1 5 10Thr Ala Ser Ala Leu Tyr Arg Glu

The information of 15 20 (2) SEQ ID NO:5:

(ⅰ) sequence signature:

(A) length: 17 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:5:Trp Val Arg Asp Ile Ile Asp Asp Phe Thr Asn Glu1 5 10Ser Ser Gln Lys Thr

The information of 15 (2) SEQ ID NO:6:

(ⅰ) sequence signature:

(A) length: 30 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:6:Lys Lys Leu Arg Arg Leu Leu Tyr Met Ile Tyr Met1 5 10Ser Gly Leu Ala Val Arg Val His Val Ser Lys Glu

The information of 15 20Glu Gln Tyr Tyr Asp Tyr25,30 (2) SEQ ID NO:7:

(ⅰ) sequence signature:

(A) length: fifteen amino acid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 1

(D) out of Memory :/annotate=" Ile, Met or Leu "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 2

(D) out of Memory :/annotate=" Ser or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 4

(D) out of Memory :/annotate=" Ile or Arg "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 5

(D) out of Memory :/annotate=" Lys or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 10

(D) out of Memory :/annotate=" His or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 11

(D) out of Memory :/annotate=" Lys or Arg "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 12

(D) out of Memory :/annotate=" Ile, Met or Leu "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 14

(D) out of Memory :/annotate=" Gly or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 15

(D) out of Memory :/annotate=" Ile, Met or Val "

(ⅹ ⅰ) sequence description: SEQ ID NO:7:Xaa Xaa Glu Xaa Xaa Gly Val Ile Val Xaa Xaa Xaa1 5 10Glu Xaa Xaa

The information of 15 (2) SEQ ID NO:8:

(ⅰ) sequence signature:

(A) length: 36 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:8:Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp1 5 10Thr Ala Ser Ala Leu Tyr Arg Glu Gly Gly Ala Gly

The information of 15 20Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys25,30 35 (2) SEQ ID NO:9:

(ⅰ) sequence signature:

(A) length: 33 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:9:Trp Val Arg Asp Ile Ile Asp Asp Phe Thr Asn Glu1 5 10Ser Ser Gln Lys Thr Gly Gly Ala Gly Cys Lys Asn

The information of 15 20Phe Phe Trp Lys Thr Phe Thr Ser Cys25,30 (2) SEQ ID NO:10:

(ⅰ) sequence signature:

(A) length: 46 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:10:Lys Lys Leu Arg Arg Leu Leu Tyr Met Ile Tyr Met1 5 10Ser Gly Leu Ala Val Arg Val His Val Ser Lys Glu

15?????????????????20Glu?Gln?Tyr?Tyr?Asp?Tyr?Gly?Gly?Ala?Gly?Cys?Lys25??????????????????30??????????????????35Asn?Phe?Phe?Trp?Lys?Thr?Phe?Thr?Ser?Cys

The information of 40 45 (2) SEQ ID NO:11:

(ⅰ) sequence signature:

(A) length: 31 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 1

(D) out of Memory :/annotate=" Ile, Met or Leu "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 2

(D) out of Memory :/annotate=" Ser or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 4

(D) out of Memory :/annotate=" Ile or Arg "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 5

(D) out of Memory :/annotate=" Lys or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 10

(D) out of Memory :/annotate=" His or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 11

(D) out of Memory :/annotate=" Lys or Arg "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 12

(D) out of Memory :/annotate=" Ile, Met or Leu "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 14

(D) out of Memory :/annotate=" Gly or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 15

(D) out of Memory :/annotate=" Ile, Met or Val "

(ⅹ ⅰ) sequence description: SEQ ID NO:11:Xaa Xaa Glu Xaa Xaa Gly Val Ile Val Xaa Xaa Xaa1 5 10Glu Xaa Xaa Gly Gly Ala Gly Cys Lys Asn Phe Phe

The information of 15 20Trp Lys Thr Phe Thr Ser Cys25,30 (2) SEQ ID NO:12:

(ⅰ) sequence signature:

(A) length: 31 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 17

(D) out of Memory :/annotate=" Ile, Met or Leu "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 18

(D) out of Memory :/annotate=" Ser or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 21

(D) out of Memory :/annotate=" Ile or Arg "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 22

(D) out of Memory :/annotate=" Lys or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 26

(D) out of Memory :/annotate=" His or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 27

(D) out of Memory :/annotate=" Lys or Arg "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 28

(D) out of Memory :/annotate=" Ile, Met or Leu "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 30

(D) out of Memory :/annotate=" Gly or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 31

(D) out of Memory :/annotate=" Ile, Met or Val "

(ⅹ ⅰ) sequence description: SEQ ID NO:12:Ala Gly Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr1 5 10Ser Cys Gly Gly Xaa Xaa Glu Ile Xaa Xaa Val Ile

The information of 15 20Val Xaa Xaa Xaa Glu Xaa Xaa25,30 (2) SEQ ID NO:13:

(ⅰ) sequence signature:

(A) length: 51 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:13:Ala Gly Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr1 5 10Ser Cys Gly Gly Trp Val Arg Asp Ile Ile Asp Asp

15??????????????????20Phe?Thr?Asn?Glu?Ser?Ser?Gln?Lys?Thr?Gly?Gly?Thr25??????????????30??????????????????35Ala?Lys?Ser?Lys?Lys?Phe?Pro?Ser?Tyr?Thr?Ala?Thr

40??????????????????45Tyr?Gln?Phe

The information of 50 (2) SEQ ID NO:14:

(ⅰ) sequence signature:

(A) length: 18 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:14:Lys Lys Lys Ile Ile Thr Ile Thr Arg Ile Ile Thr1 5 10Ile Ile Thr Thr Ile Asp

The information of 15 (2) SEQ ID NO:15:

(ⅰ) sequence signature:

(A) length: fifteen amino acid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:15:Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile Pro Gln1 5 10Ser Leu Asp

The information of 15 (2) SEQ ID NO:16:

(ⅰ) sequence signature:

(A) length: 36 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:16:Gly Ile Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile1 5 10Thr His Val Asp Thr Glu Ser Tyr Gly Gly Ala Gly

The information of 15 20Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys25,30 35 (2) SEQ ID NO:17:

(ⅰ) sequence signature:

(A) length: 38 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:17:Thr Ala Lys Ser Lys Lys Phe Pro Ser Tyr Thr Ala1 5 10Thr Tyr Gln Phe Gly Ile Leu Glu Ser Arg Gly Ile

The information of 15 20Lys Ala Arg Ile Thr His Val Asp Thr Glu Ser Tyr25,30 35Gly Gly (2) SEQ ID NO:18:

(ⅰ) sequence signature:

(A) length: 20 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:18:Lys Lys Lys Phe Phe Leu Leu Thr Arg Ile Leu Thr1 5 10Ile Pro Gln Ser Leu Asp Gly Gly

The information of 15 20 (2) SEQ ID NO:19:

(ⅰ) sequence signature:

(A) length: 38 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:19:Thr Ala Lys Ser Lys Lys Phe Pro Ser Tyr Thr Ala1 5 10Thr Tyr Gln Phe Gly Gly Lys Lys Lys Phe Phe Leu

The information of 15 20Leu Thr Arg Ile Leu Thr Ile Pro Gln Ser Leu Asp25,30 35Gly Gly (2) SEQ ID NO:20:

(ⅰ) sequence signature:

(A) length: 34 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:20:Lys Lys Lys Ile Ile Thr Ile Thr Arg Ile Ile Thr1 5 10Ile Ile Thr Thr Ile Asp Gly Gly Ala Gly Cys Lys

The information of 15 20Asn Phe Phe Trp Lys Thr Phe Thr Ser Cys25,30 (2) SEQ ID NO:21:

(ⅰ) sequence signature:

(A) length: 20 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:21:Gly Ile Leu Glu Ser Arg Gly Ile Lys Ala Arg Ile1 5 10Thr His Val Asp Thr Glu Ser Tyr

The information of 15 20 (2) SEQ ID NO:22:

(ⅰ) sequence signature:

(A) length: 36 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:22:Ala Gly Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr1 5 10Ser Cys Gly Gly Gly Ile Leu Glu Ser Arg Gly Ile

The information of 15 20Lys Ala Arg Ile Thr His Val Asp Thr Glu Ser Tyr25,30 35 (2) SEQ ID NO:23:

(ⅰ) sequence signature:

(A) length: 25 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:23:Ala Leu Asn Ile Trp Asp Arg Phe Asp Val Phe Ser1 5 10Thr Leu Gly Ala Thr Ser Gly Tyr Leu Lys Gly Asn

The information of 15 20Ser25 (2) SEQ ID NO:24:

(ⅰ) sequence signature:

(A) length: 41 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:24:Ala Leu Asn Ile Trp Asp Arg Phe Asp Val Phe Ser1 5 10Thr Leu Gly Ala Thr Ser Gly Tyr Leu Lys Gly Asn

15??????????????????20Ser?Gly?Gly?Ala?Gly?Cys?Lys?Asn?Phe?Phe?Trp?Lys25??????????????????30??????????????????35Thr?Phe?Thr?Ser?Cys

The information of 40 (2) SEQ ID NO:25:

(ⅰ) sequence signature:

(A) length: 54 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:25:Thr Ala Lys Ser Lys Lys Phe Pro Ser Tyr Thr Ala1 5 10Thr Tyr Gln Phe Gly Gly Ser Asp Phe Phe Pro Ser

15??????????????????20Val?Arg?Asp?Leu?Leu?Asp?Thr?Ala?Ser?Ala?Leu?Tyr25??????????????????30??????????????????35Arg?Glu?Gly?Gly?Ala?Gly?Cys?Lys?Asn?Phe?Phe?Trp

40??????????????????45Lys?Thr?Phe?Thr?Ser?Cys

The information of 50 (2) SEQ ID NO:26:

(ⅰ) sequence signature:

(A) length: 54 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:26:Ala Gly Cys Lys Asn Phe Phe Trp Lys Thr Phe Thr1 5 10Ser Cys Gly Gly Ser Asp Phe Phe Pro Ser Val Arg

15??????????????????20Asp?Leu?Leu?Asp?Thr?Ala?Ser?Ala?Leu?Tyr?Arg?Glu25??????????????????30??????????????????35Gly?Gly?Thr?Ala?Lys?Ser?Lys?Lys?Phe?Pro?Ser?Tyr

40??????????????????45Thr?Ala?Thr?Tyr?Gln?Phe

The information of 50 (2) SEQ ID NO:27:

(ⅰ) sequence signature:

(A) length: 12 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: the information of SEQ ID NO:27:Thr Ile Asn Lys Pro Lys Gly Tyr Val Gly Lys Glu1 5 10 (2) SEQ ID NO:28:

(ⅰ) sequence signature:

(A) length: 22 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:28:Asp Ser Glu Thr Ala Asp Asn Leu Glu Lys Thr Val1 5 10Ala Ala Leu Ser Ile Leu Pro Gly His Gly

The information of 15 20 (2) SEQ ID NO:29:

(ⅰ) sequence signature:

(A) length: fifteen amino acid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:29:Leu Ser Glu Ile Lys Gly Val Ile Val His Arg Leu1 5 10Glu Gly Val

The information of 15 (2) SEQ ID NO:30:

(ⅰ) sequence signature:

(A) length: fifteen amino acid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 2

(D) out of Memory :/annotate=" Ser or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 5

(D) out of Memory :/annotate=" Lys or Arg "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 6

(D) out of Memory :/annotate=" Gly or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 10

(D) out of Memory :/annotate=" His or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 11

(D) out of Memory :/annotate=" Lys or Arg "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 14

(D) out of Memory :/annotate=" Gly or Thr "

(ⅹ ⅰ) sequence description: SEQ ID NO:30:Ile Xaa Glu Ile Xaa Xaa Val Ile Val Xaa Xaa Ile1 5 10Glu Xaa Ile

The information of 15 (2) SEQ ID NO:31:

(ⅰ) sequence signature:

(A) length: 19 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 4

(D) out of Memory :/annotate=" Ser or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 7

(D) out of Memory :/annotate=" Lys or Arg "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 8

(D) out of Memory :/annotate=" Gly or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 12

(D) out of Memory :/annotate=" His or Thr "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 13

(D) out of Memory :/annotate=" Lys or Arg "

(ⅸ) characteristic:

(A) title/key word: decorating site

(B) position: 16

(D) out of Memory :/annotate=" Gly or Thr "

(ⅹ ⅰ) sequence description: SEQ ID NO:31:Ile Ser Ile Xaa Glu Ile Xaa Xaa Val Ile Val Xaa1 5 10Xaa Ile Glu Xaa Ile Leu Phe

The information of 15 (2) SEQ ID NO:32:

(ⅰ) sequence signature:

(A) length: 17 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:32Lys Lys Gln Tyr Ile Lys Ala Asn Ser Lys Phe Ile1 5 10Gly Ile Thr Glu Leu

The information of 15 (2) SEQ ID NO:33:

(ⅰ) sequence signature:

(A) length: 22 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:33:Lys Lys Phe Asn Asn Phe Thr Val Ser Phe Trp Leu1 5 10Arg Val Pro Lys Val Ser Ala Ser His Leu

The information of 15 20 (2) SEQ ID NO:34:

(ⅰ) sequence signature:

(A) length: 27 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:34:Tyr Asp Pro Asn Tyr Leu Arg Thr Asp Ser Asp Lys1 5 10Asp Arg Phe Leu Gln Thr Met Val Lys Leu Phe Asn

The information of 15 20Arg Ile Lys25 (2) SEQ ID NO:35:

(ⅰ) sequence signature:

(A) length: 24 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:35:Gly Ala Tyr Ala Arg Cys Pro Asn Gly Thr Arg Ala1 5 10Leu Thr Val Ala Glu Leu Arg Gly Asn Ala Glu Leu

The information of 15 20 (2) SEQ ID NO:36:

(ⅰ) sequence signature:

(A) length: 21 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:36:Val Ser Phe Gly Val Trp Ile Arg Thr Pro Pro Ala1 5 10Tyr Arg Pro Pro Asn Ala Pro Ile Leu

The information of 15 20 (2) SEQ ID NO:37:

(ⅰ) sequence signature:

(A) length: 20 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:37:Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys1 5 10Trp Gly Glu Leu Met Thr Leu Ala

The information of 15 20 (2) SEQ ID NO:38:

(ⅰ) sequence signature:

(A) length: 19 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:38:Arg Ala Gly Arg Ala Ile Leu His Ile Pro Thr Arg1 5 10Ile Arg Gln Gly Leu Glu Arg

The information of 15 (2) SEQ ID NO:39:

(ⅰ) sequence signature:

(A) length: 21 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:39:Ala Val Ala Glu Gly Thr Asp Arg Val Ile Glu Val1 5 10Leu Gln Arg Ala Gly Arg Ala Ile Leu

The information of 15 20 (2) SEQ ID NO:40:

(ⅰ) sequence signature:

(A) length: 39 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:40:Glu Glu Ile Val Ala Gln Ser Ile Ala Leu Ser Ser1 5 10Leu Met Val Ala Gln Ala Ile Pro Leu Val Gly Glu

The information of 15 20Leu Val Asp Ile Gly Phe Ala Ala Thr Asn Phe Val25,30 35Glu Ser Cys (2) SEQ ID NO:41:

(ⅰ) sequence signature:

(A) length: 21 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:41:Asp Ile Glu Lys Lys Ile Ala Lys Met Glu Lys Ala1 5 10Ser Ser Val Phe Asn Val Val Asn Ser

The information of 15 20 (2) SEQ ID NO:42:

(ⅰ) sequence signature:

(A) length: 17 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:42:Lys Trp Phe Lys Thr Asn Ala Pro Ash Gly Val Asp1 5 10Glu Lys Ile Arg Ile

The information of 15 (2) SEQ ID NO:43:

(ⅰ) sequence signature:

(A) length: 14 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: the information of SEQ ID NO:43:Gly Leu Gln Gly Lys Ile Ala Asp Ala Val Lys Ala1 5 10Lys Gly (2) SEQ ID NO:44:

(ⅰ) sequence signature:

(A) length: 19 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQID NO:44:Gly Leu Ala Ala Gly Leu Val Gly Met Ala Ala Asp1 5 10Ala Met Val Glu Asp Val Asn

The information of 15 (2) SEQ ID NO:45:

(ⅰ) sequence signature:

(A) length: 20 aminoacid

(B) type: aminoacid

(D) topological structure: linearity

(ⅱ) molecule type: peptide

(ⅹ ⅰ) sequence description: SEQ ID NO:45:Ser Thr Glu Thr Gly Asn Gln His His Tyr Gln Thr1 5 10Arg Val Val Ser Asn Ala Asn Lys

15??????????????????20

Claims (29)

1. a peptide conjugates contains the helper T cell epitope sequence (Th) that is covalently bonded in somatostatin or its cross reactivity and immunologic function analog.

2. the peptide conjugates of claim 1, wherein said peptide conjugates is expressed from the next:

H ₂N-(A) _n-(somatostatin peptides)-(B) _o-(Th) _m-X

Or

H ₂N-(A) _n-(Th) _m-(B) _o-(somatostatin peptides)-X is wherein

H ₂N is the terminal α-NH of the N-of peptide conjugates ₂,

Each A is a seed amino acid or a kind of general immunostimulatory sequence independently;

Each B be selected from aminoacid ,-NHCH (X) CH ₂SCH ₂CO-,-NHCH (X) CH ₂SCH ₂CO (Lys-of ε-N) ,-NHCH (X) CH ₂The S-succinimido (Lys-of ε-N) and-NHCH (X) CH ₂S-(succinimido)-;

Each Th is independently for containing aminoacid sequence or its immunostimulant analog or the fragment of helper T cell epitope;

Somatostatin peptides is somatostatin or its cross reactivity and immunologic function analog;

X is a kind of amino acid whose α-COOH or α-CONH ₂

N is 1 to about 10;

M is 1 to about 4; With

O is 0 to about 10.

3. the peptide conjugates of claim 1, wherein said peptide conjugates is expressed from the next:

H ₂N-(somatostatin peptides)-(B) _o-(Th) _m-(A) n-X

Or

H ₂N-(Th) _m-(B) _o-(somatostatin peptides)-(A) _n-X wherein

H ₂N is the terminal α-NH of the N-of peptide conjugates ₂,

Each A is a seed amino acid or a kind of general immunostimulatory sequence independently;

Each B be selected from aminoacid ,-NHCH (X) CH ₂SCH ₂CO-,-NHCH (X) CH ₂SCH ₂CO (Lys-of ε-N) ,-NHCH (X) CH ₂The S-succinimido (Lys-of ε-N) and-NHCH (X) CH ₂S-(succinimido)-;

Each Th is independently for containing aminoacid sequence or its immunostimulant analog or the fragment of helper T cell epitope;

Somatostatin peptides is somatostatin or its cross reactivity and immunologic function analog;

X is a kind of amino acid whose α-COOH or α-CONH ₂

N is 1 to about 10;

M is 1 to about 4; With

O is 0 to about 10.

4. claim 2 or 3 peptide conjugates, wherein each B is selected from natural amino acid or alpha-non-natural amino acid.

5. the peptide conjugates of one of claim 1-4, wherein said somatostatin peptides is a somatostatin.

6. the peptide conjugates of one of claim 1-4, wherein said Th is a kind of SSAL epi-position.

7. the peptide conjugates of one of claim 1-4, wherein said Th has the aminoacid sequence that is selected from SEQ IDNO:4, EQ ID NO:5, SEQID NO:6, SEQ ID NO:7, SEQ ID NO:14, SEQID NO:15, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:27, SEQ IDNO:28 and SEQ ID NO:29 SEQ ID NO:30 and SEQ ID NO:31.

8. the peptide conjugates of claim 2, wherein said peptide conjugates has the aminoacid sequence that is selected from SEQ IDNO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQID NO:19, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ IDNO:25 and SEQ ID NO:26.

9. claim 2 or 3 peptide conjugates, wherein at least one A is a kind of invasin domain.

10. the peptidic constructs of claim 2, wherein n is 3, (A) ₃Be (invasin domain)-Gly-Gly.

11. the peptide conjugates of claim 9 or 10, wherein said invasin has the aminoacid sequence of SEQ ID NO:2.

12. one kind has 25 approximately to about 90 amino acid whose synthetic peptides, it contains following amino acid sequences:

(a) a kind of invasin domain,

(b) a kind of helper T cell (Th) epi-position and

(c) somatostatin or its cross reactivity and immunologic function analog.

13. peptide that contains the aminoacid sequence that is selected from SEQ ID NO:17, SEQ ID NO:19 and SEQ ID NO:25.

14. peptide of one of claim 1 to 13 or peptide conjugates, wherein said peptide stimulate the immunne response of mammal generation to somatostatin.

15. the peptide of claim 14 or peptide conjugates wherein cause somatostatin level minimizing in the described mammal with described peptide conjugates to mammiferous immunity.

16. a pharmaceutical composition contains peptide or the peptide conjugates of a kind of one of claim 1-15 of immune effective dose and a kind of drug acceptable carrier.

17. the pharmaceutical composition of claim 16, the described immune effective dose of wherein said peptide or peptide conjugates are every dosage per kilogram of body weight, and about 0.5 μ g arrives about 1 mg.

18. a method of inducing mammal to produce anti--somatostatin antibody comprises the pharmaceutical composition to described administration claim 16 or 17.

19. a method that improves the mammal growth rate comprises the pharmaceutical composition to described administration claim 16 or 17.

20. a method that improves the mammal growth rate comprises to administration dosage being enough to reduce the claim 16 of somatostatin level or 17 pharmaceutical composition.

21. the compositions of the mixture of peptide that contains two or more claim 1-15 or peptide conjugates.

22. a pharmaceutical composition, it contains a kind of compositions and a kind of medicine acceptable carrier of claim 21 of immune effective dose.

23. the pharmaceutical composition of claim 22, the described immune effective dose of wherein said compositions are every dosage per kilogram of body weight, and about 0.5 μ g arrives about 1 mg.

24. a method of inducing mammal to produce somatostatin antibody comprises the pharmaceutical composition to described administration claim 22 or 23.

25. a method that improves the mammal growth rate comprises the pharmaceutical composition to described administration claim 22 or 23.

26. a method that improves growth rate comprises to administration dosage being enough to reduce the claim 22 of somatostatin level or 23 pharmaceutical composition.

27. one kind contains a lysine, three lysines or seven covalently bound respectively branched polymers that the peptide conjugates of one of two, four or eight claim 1-15 is arranged of lysine core.

28. one or more peptide conjugates that contain one of claim 1-3 and claim 5-15, by the crosslinked polymer of bi-functional cross-linking agent.

29. Th epitope peptide that is selected from SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ IDNO:7, SEQ ID NO:14, SEQID NO:15, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:27, SEQ ID NO:28 and SEQ ID NO:29, SEQ ID NO:30 and SEQ ID NO:31.