CN101421297A

CN101421297A - Peptides that block the binding of igg to fcrn

Info

Publication number: CN101421297A
Application number: CNA2007800126775A
Authority: CN
Inventors: 亚当·R·梅佐; 凯文·A·麦克唐奈; 克里斯蒂娜·T·A·赫希尔; 艾尔弗雷多·卡斯特罗
Original assignee: Syntonix Pharmaceuticals Inc
Current assignee: Bioverativ Therapeutics Inc
Priority date: 2006-02-17
Filing date: 2007-02-16
Publication date: 2009-04-29
Also published as: ZA200807631B

Abstract

The invention relates to peptides which bind to human FcRn and inhibit binding of the Fc portion of an IgG to an FcRn, thereby modulating serum IgG levels. The disclosed compositions and methods may be used for example, in treating autoimmune diseases and inflammatory disorders. The invention also relates to methods of using and methods of making the peptides of the invention.

Description

Blocking-up IgG is to the bonded peptide of FcRn

In first to file

The application requires the right of priority of U.S. Provisional Application of submitting on February 17th, 2,006 60/774,853 and the U.S. Provisional Application of submitting on June 23rd, 2,006 60/805,634, at this their full contents is incorporated herein by reference.

Invention field

Generally speaking, the present invention relates to immunoregulation agent field.In particular, the present invention relates to peptide, it can combine and suppress the combination of FcRn to immunoglobulin G (IgG) Fc part with Fc newborn infant's acceptor (FcRn), regulate and control the serum IgG level thus.The invention further relates to the active peptide adjusting control agent of FcRn, contain the medicine of described peptide, and by using the method that described medicine treats for the experimenter disease and illness, described peptide adjusting control agent can stop FcRn to bring into play function in relating to the cell mechanism of keeping IgG level in the serum, and described disease and illness can be alleviated by reducing the serum IgG level.

Background of invention

The abundantest antibody isotype is IgG in the serum, its mediation at the protection of pathogenic agent in and promote the immunity system composition to raise in the allergy of tissue, mucous membrane and skin surface or the inflammatory response in mediation to have keying action.Junghans, Immunologic Research 16 (1): 29 (1997). and in addition, IgG also is the key component of various autoimmune disease.

Under normal operation, IgG is long for the serum half-life of other plasma proteins in the transformation period in the serum.For example, the serum half-life of IgG is 5 to 7 days in mouse, and is 22-23 days in the people.Roopenian et al., J.Immunology 170:3528 (2003); The long half-lift of Junghans andAnderson, Proc.Natl.Acad.Sci USA 93:5512 (1996) .IgG this partly be because it is the combination of FcRn to the Fc acceptor.FcRn is bonded to the constant region of IgG, i.e. the Fc known to.Though FcRn is characterized as being the newborn infant of parent IgG at first and transports acceptor, it also brings into play the function that protection IgG avoids degrading in the adult.FcRn is bonded to by the IgG of pinocytosis and protects it to avoid the lysosomal effect of reduction property, makes its recirculation get back to the extracellular compartment then.Junghans and Anderson, Proc.Natl.Acad.Sci.USA 93:5512 (1996); Roopenian et al., J.Immunology 170:3528 (2003) if. the concentration of IgG reaches the level that exceeds available FcRn, and so unconjugated IgG can not be protected and avoid degradation property mechanism, and therefore has short serum half-life.Brambell et al., Nature 203:1352 (1964). in addition,, it is believed that many FcRn link in cell and with endoplasm vesica film though FcRn is expressed on the cell surface, and after IgG gone into cell by endocytosis, the interaction between IgG and the FcRn takes place in born of the same parents.

Structurally, FcRn exists as heterodimer, is made of a light chain and a non-covalent bonded heavy chain that is called the α chain that is called the β chain.The FcRn light chain is as β ₂-microglobulin (β ₂M) and more well known, it also is the composition of major histocompatibility complex I (MHC I).The α chain of FcRn is the protein of 46kD, and it constitutes: be divided into three subdomain α ₁, α ₂And α ₃Ectodomain; Stride the film district; Duan kytoplasm tail relatively.Burmeister?et?al.，Nature?372：336(1994).

FnRn identifies first that in the neonate rat intestines its performance mediation absorbs IgG antibody and promotes it to be transported to the function of the recycle system from breast milk there.Leach et al., J.Immunology157:3317 (1996). also isolated FcRn from people's placenta, its mediates parent IgG absorption and is transported to fetal circulation there.In the adult, FcRn expresses (U.S. Patent No. 6,030 in epithelium, 613 and 6,086,875), closely manage epithelium (Kobayashi et al., Am.J.Physiol. (2002) such as, but not limited to lung (Israel et al., Immunology 92:69 (1997)), intestines and kidney; RenalPhysiol.282:F358 (2002)) and nose, vagina and courage system surface.In addition, FcRn has hinted its importance in the IgG stable state generally expressing on the endotheliocyte.Ward?et?al.，InternationalImmunology?15(2)：187(2002)；Ghetie?et?al.，Eur.J.Immunology?26：690(1996).

Usually, FcRn brings into play function thus by the katabolism that the Fc that is bonded to IgG partly comes antagonism IgG in the IgG stable state.In case by endocytosis, IgG is trapped in the endocytic vacuole, beginning is merged with acid elementary endosome.Crystallographic Study has hinted that the stoichiometry of FcRn-IgG complex body constitutes two molecule FcRn to a part IgG (Burmeister et al., Nature 372:336 (1994)), and think that the combination of these two kinds of molecules occurs in that the Fc near CH2 and CH3 structural domain interface partly goes up (Burmeister et al., Nature 372:379 (1994)) among the IgG.The endosome fusion event has been represented a stage of lysosome degradation pathway, and it will be included in the complex biological molecular degradation in the endosome or be decomposed into the composition composition.The low pH environment of elementary endosome promotes the combination of FcRn to IgG, and any by the antigenic release of IgG institute bonded.Therefore, antigen is degraded, and the FcRn-IgG complex body is avoided degraded and finally is recycled to cell surface, and the physiological pH of extracellular environment promotes to discharge IgG from FcRn there.

In order to study the contribution of FcRn to the IgG stable state, transformed mouse in case " knocking out " to small part coding β ₂The gene of m and FcRn heavy chain makes these protein not express.WO 02/43658; Junghans andAnderson, Proc.Natl.Acad.Sci.USA 93:5512 (1996). in these two kinds of knock-out mice strains, transformation period and the concentration of IgG in serum all sharply reduces, and has hinted the FcRn dependency mechanism that relates to the IgG stable state.

Suppress IgG and be bonded to FcRn by stoping IgG recirculation reduction IgG serum half-life.Therefore, blocking-up or antagonism IgG can be used to regulate, treat or prevent to involve the method for immunoreactive illness to the bonded medicament of FcRn, such as for example being autoimmunity and the inflammatory diseases and the illness of feature with the IgG antibody that has improper expression.The example that blocking-up IgG Fc is bonded to the method for FcRn comprises the blocking antibody that generates FcRn.In fact, use FcRn heavy chain knock-out mice strain to generate and to block FcRn and IgG bonded antibody (WO 02/43658).Recently, identified the peptide that can be bonded to the FcRn complex body.Kolonin et al., Proc.Natl.Acad.Sci.USA 99 (20): 13055-60 (2002); U.S. Patent No. 6,212, yet 022., now still needing other medicament to regulate, treat or prevent with the immune response is illness, disease and the illness of feature.

Summary of the invention

Thereby, the invention provides peptide, it can specificity be bonded to FcRn and suppress the combination of IgG Fc to FcRn, thus by stoping FcRn to stop IgG recirculation in the performance function on that its protection IgG avoids the lysosome degraded.In exemplary embodiment, IgG1, IgG2, IgG3 or IgG4 subclass that described peptide is bonded to FcRn and inhibition Fc are bonded to FcRn.

In some embodiments, peptide of the present invention comprises sequence:

-Gly-X ₆-X ₇-X ₈-X ₉-X ₁₀-X ₁₁-

Wherein:

-X ₆Be selected from positively charged amino acid, die aromatischen Aminosaeuren, positively charged die aromatischen Aminosaeuren and their analogue;

-X ₇Be selected from phenylalanine and phenylalanine analogues;

-X ₈And X ₉Be selected from glycine, sarkosine, aspartic acid, D-amino acid, α-An Jiyidingsuan and their analogue, perhaps X independently of one another ₈With X ₉One time-out forms dipeptide analogue;

-X ₁₀Be selected from amino acid and their analogue, perhaps X ₁₀With X ₉One time-out forms dipeptide analogue;

-X ₁₁Be selected from tyrosine and tyrosine analogue.

Perhaps, peptide of the present invention can comprise sequence:

R ₁-Gly-X ₆-X ₇-X ₈-X ₉-X ₁₀-X ₁₁-R ₂

Wherein:

-R ₁Has general formula X ₁-X ₂-X ₃-X ₄-

Wherein

-X ₁Be selected from hydrogen, acyl group and amino acid blocking group;

-X ₂Disappearance or to be selected from amino acid, length be 2-15 amino acid whose peptide and their analogue;

-X ₃Disappearance or energy and X ₁₀, X ₁₂Or X ₁₃Form the amino acid of bridge (bridge) or their analogue, wherein said bridge be selected from N-terminal to the bridge of C-terminal, side chain to skeleton bridge and side chain to the bridge of side chain; And

-X ₄Disappearance or to be selected from amino acid, length be 2-15 amino acid whose peptide and their analogue;

-X ₆,-X ₇,-X ₈,-X ₉,-X ₁₀With-X ₁₁With above definition, and

-R ₂Has general formula-X ₁₂-X ₁₃-X ₁₄-X ₁₅

Wherein

-X ₁₂Disappearance or amino acid or their analogue;

-X ₁₃Disappearance or amino acid or their analogue;

-X ₁₄Disappearance or to be selected from amino acid, length be 2-15 amino acid whose peptide and their analogue; And

-X ₁₅Be amino group or carboxy protective group.

Typically, the length of peptide of the present invention is at least 7, nearly 50 amino acid.Peptide of the present invention can be used as polymer and exists, such as for example dimer, tripolymer or the tetramer.In some embodiments, peptide of the present invention can be more responsive to pinosome, makes the rapider combination of this peptide, and therefore by kidney drainage still less.The invention further relates to medicinal compositions, it comprises one or more peptides of the present invention.

Peptide of the present invention can comprise:

A) be selected from down the aminoacid sequence of organizing:

QRFCTGHFGGLYPCNGP(SEQ?ID?NO：1)，

GGGCVTGHFGGIYCNYQ(SEQ?ID?NO：2)，

KIICSPGHFGGMYCQGK(SEQ?ID?NO：3)，

PSYCIEGHIDGIYCFNA (SEQ ID NO:4) and

NSFCRGRPGHFGGCYLF(SEQ?ID?NO：5)；

B) with SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 and SEQ IDNO:5 in one or more substantially the same aminoacid sequence;

C) with SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 and SEQ IDNO:5 in one or more at least 80% identical aminoacid sequence;

D) by one, two, three, four or five deletions, substitute or add SEQID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 or the SEQ ID NO:5 that sudden change is modified;

E) by at least one amino acid whose SEQ ID NO:1, SEQ ID NO:2, SEQID NO:3, SEQ ID NO:4 or SEQ ID NO:5 that modifies that substitute, wherein with the naturally occurring amino acid replacement of described at least one amino acid;

F), wherein described at least one amino acid is substituted with D-amino acid and their analogue by at least one amino acid whose SEQ ID NO:1, SEQ ID NO:2, SEQ IDNO:3, SEQ ID NO:4 or SEQ ID NO:5 that modifies that substitute;

G) by at least one amino acid whose SEQ ID NO:1, SEQ ID NO:2, SEQID NO:3, SEQ ID NO:4 or SEQ ID NO:5 that modifies that substitute, wherein with described at least one amino acid with the N-amino acid replacement that methylates;

H) by at least one amino acid whose SEQ ID NO:1, SEQ ID NO:2, SEQID NO:3, SEQ ID NO:4 or SEQ ID NO:5 that modifies that substitute, wherein with the amino acid replacement of described at least one amino acid with the non-natural existence; With

I), wherein described at least one amino acid is substituted with the amino acid analog thing by at least one amino acid whose SEQ ID NO:1, SEQ ID NO:2, SEQ IDNO:3, SEQ ID NO:4 or SEQ ID NO:5 that modifies that substitute.

The invention further relates to the method for regulating the IgG level in experimenter's serum, comprise composition to experimenter's administering therapeutic significant quantity, said composition comprises one or more peptides of the present invention, and this peptide can be bonded to FcRn and stop FcRn to be bonded to the Fc part of IgG molecule.In certain embodiments, method of the present invention is used for reducing the transformation period of the solubility IgG of experimenter's serum.Using composition results of the present invention is: compare with the transformation period of solubility IgG in experimenter's serum before using described peptide, the transformation period of the solubility IgG in experimenter's serum reduces.

The present invention further provides the method that the Fc that suppresses human IgG partly is bonded to FcRn, compared reduction for the serum-concentration of FcRn bonded IgG Fc part and the IgG serum-concentration before the treatment with realization.The method that reduces the IgG serum-concentration comprises: to the composition of experimenter's administering therapeutic significant quantity, said composition comprises one or more peptides of the present invention, and this peptide can be bonded to FcRn and stop FcRn to be bonded to the Fc part of IgG molecule.In one embodiment, the human IgG serum-concentration is reduced at least 5%, the reduction such as at least 15%, the perhaps reduction of human IgG serum-concentration at least 25%.

One embodiment of the invention provide treatment to suffer from the experimenter's of at least a autoimmune disorder method, comprise composition to experimenter's administering therapeutic significant quantity, said composition comprises one or more peptides of the present invention, and this peptide can be bonded to FcRn and stop FcRn to be bonded to the Fc part of IgG molecule.An alternative embodiment of the present invention provides treatment to suffer from the experimenter's of at least a inflammatory conditions method, by composition to experimenter's administering therapeutic significant quantity, said composition comprises one or more peptides of the present invention, and this peptide can be bonded to FcRn and stop FcRn to be bonded to the Fc part of IgG molecule.In other embodiment, method of the present invention can be used to prevent, treatment or metering needle be to the immunne response of therapeutic protein or gene therapy vector.

Other embodiment of the present invention, purpose and advantage are partly listed in ensuing specification sheets, and will be clearly according to specification sheets partly, perhaps can know by enforcement of the present invention.By key element and the combination of in claims, specifically noting, can be familiar with these embodiments of the present invention, purpose and advantage.

Should be appreciated that for desired invention, above generality describe and hereinafter all only be exemplary in detail with indicative, and nonrestrictive.

Brief Description Of Drawings

Fig. 1 has shown people's total length FcRn and people's beta-2 microglobulin (β ₂The cDNA sequence of reading frame m) (ORF).

Fig. 2 has shown the synthetic summary of the terminal aldehyde peptide monomer of N-(SEQ ID NO:319).

Fig. 3 has shown the summary of synthesizing peptide dimer by standard reductive alkylation.The synthetic example that shows explaining property of peptide No.270.

Fig. 4 has shown by use and has contained the summary that the peptide of the peptide of two mercaptan joints and acetobromization synthesizes peptide dimer.The synthetic example that shows explaining property of peptide No.100.Exist (the SEQ ID NO:320) of bridge indicated in the bracket that is placed on the horizontal direction of peptide sequence top.

Fig. 5 has shown that use contains the peptide of mercaptan joint and the peptide of acetobromization synthesizes peptide dimer.The synthetic example that shows explaining property of peptide No.122.Exist (the SEQ ID NO:320) of bridge indicated in the bracket that is placed on the horizontal direction of peptide sequence top.

Fig. 6 has shown and uses the joint that contains diprotic acid to synthesize peptide dimer.The synthetic example that shows explaining property of peptide No.283.Exist (the SEQID NO:321) of bridge indicated in the bracket that is placed on the horizontal direction of peptide sequence below.

Fig. 7 has shown and uses the joint that contains amine to synthesize peptide dimer.The synthetic example that shows explaining property of peptide No.280.Exist (the SEQ IDNO:321) of bridge indicated in the bracket that is placed on the horizontal direction of peptide sequence below.

Fig. 8 has shown that use peptide aldehyde and protein C ysFc synthesize peptide-Fc fusions.The synthetic example that shows explaining property of peptide No.252-Fc.The existence of bridge indicated in the bracket that is placed on the horizontal direction of peptide sequence below.

Fig. 9 has shown that the catabolic kinetics of human IgG in the TG32B mouse (transforms mouse and come expressing human FcRn and people β ₂M, rather than mouse FcRn or β ₂M).

Figure 10 has shown that behind intravenous injection peptide No.270, the katabolism kinetics of the biotinylation human IgG in the macaque has been quickened.

Figure 11 has shown behind intravenous injection peptide No.270, the serum level of the endogenous IgG in the macaque.

Figure 12 is the representative of the result shown in Figure 11, has wherein made endogenous macaque IgG level with respect to T ₀Horizontal normalizing.

After Figure 13 has shown intravenous injection peptide No.270, the level of the endogenous serum albumin in the macaque.

After Figure 14 has shown intravenous injection peptide No.270, the level of the endogenous IgM in the macaque.

After Figure 15 has shown intravenous injection peptide No.231, peptide No.274 and peptide No.252-Fc, the catabolic kinetics of the human IgG in the TG32B mouse.

After Figure 16 has shown intravenous injection peptide No.270, the catabolic kinetics of the human IgG in the TG32B mouse.

After Figure 17 has shown intravenous injection peptide No.283, the catabolic kinetics of the human IgG in the TG32B mouse.

Figure 18 has shown the molecular weight of peptide No.289, records by the peptide No.289 with SDS-PAGE analysis purifying on the Tris-Gly of 4-20% gel.First road contains molecular weight marker.Second road contains non-link coupled PEG _30kDaParent material.The 3rd road contains the crude product reaction mixture.The 4th road contains the peptide No.289 of purifying.

Figure 19 has shown behind intravenous injection peptide No.289, the catabolic kinetics of the human IgG in the TG32B mouse.

Figure 20 has shown behind intravenous injection 5mg/kg peptide No.283, the catabolic kinetics of biotinylation human IgG in the macaque.

Figure 21 has shown behind intravenous injection 1mg/kg peptide No.283, the catabolic kinetics of biotinylation human IgG in the macaque.

Figure 22 has shown the influence of peptide No.283 (5mg/kg) to the IgG concentration in the macaque.

Figure 23 has shown the influence of peptide No.283 (1mg/kg) to the IgG concentration in the macaque.

Figure 24 shown peptide No.283 (5mg/kg, 3 */wk, iv) to the influence of the white protein concentration in the macaque.

The description of embodiment

I. definition

" avidity " refers to the interactional feature of associativity between two biologically active molecules, the interactional intensity of its indication associativity.Affine force measurement is reported as dissociation constant (K _D), it is under defined terms, biologically active molecules begins no longer generally to be reported as nM, pM or fM in conjunction with (dissociating) concentration to its binding partners in containing the solution of biologically active molecules.Avidity intensity and K _DValue is inversely proportional to.

Term used herein " amino acid " refers to contain the compound of hydroxy-acid group and amino group.For example, amino acid can have general structure H ₂N-[C (R) (R ')] _n-C (O) OH, wherein n is the integer more than or equal to 1, and R and R ' be independently selected from hydrogen and amino acid side chain, and wherein R and R ' can form carbocyclic ring or heterocycle together.For example, when n equals 1, general formula H ₂N-[C (R) (R ')]-amino acid of C (O) OH is alpha amino acid, and when n equals 2, general formula H ₂N-C (R ₁) (R ₁')-C (R ₂) (R ₂')-amino acid of C (O) OH is beta amino acids, R wherein ₁, R ₁', R ₂And R ₂' be selected from amino acid side chain independently of one another, and wherein R and R ' or R ₂And R ₂' can form carbocyclic ring or heterocycle together.Term used herein " amino-acid residue " refers to the amino acid as a peptide or a proteinic part, and has general formula-N (H)-[C (R) (R ')] _n-C (O)-.Term used herein " amino acid side chain " refers to from natural existence or the amino acid whose any side chain of synthetic.For example, methyl can be called the L-Ala side chain, and 2-amino-1-ethyl can be called the side chain of 2,4-diamino-butanoic.

Amino acid can have R or S chirality at any chiral atom place.If the α amino-carbon is a chirality, usage charges Xi Er rule (Fischer convention) is specified the chirality of the α amino-carbon of alpha amino acid so, for L-or be D-." D-amino acid " is the amino acid that the D configuration is arranged at α carbon place.When not pointing out clear and definite configuration, those skilled in the art are interpreted as L-amino acid with amino acid.Amino acid described herein also can be racemize, non-racemize and diastereomeric form of mixtures.

Exemplary amino acid can be selected from 20 kinds of coded amino acids and their analogue, also can be selected from for example other a-amino acid, beta-amino acids, gamma-amino acid, δ-amino acid and omega-amino acid.The known undoded amino acid in peptide field, such as those at M.Bodanszky, Principles of Peptide Synthesis, 1st and 2nd revised ed., Springer-Verlag, New York, N.Y., (1984) and (1993) and Stewart and Young, Solid Phase Peptide Synthesis, 2nd ed., Pierce Chemical Co., Rockford, III. describes in (1984).Coding and undoded amino acid and amino acid analogue can be commercial available from for example Novabiochem, Bachem, Sigma Chemical Co., AdvancedChemtech; Or use means known in the art synthetic.

Amino acid can be selected from for example L-Ala, Beta-alanine, α-An Jijiersuan, the 2-aminobutyric acid, the 4-aminobutyric acid, 1-aminocyclopentanecarboxylic acid, 6-aminocaprolc acid, the 2-diaminopimelic acid, the 7-aminoheptylic acid, the 2-aminoisobutyric acid, aminomethyl pyrroles carboxylic acid, 8-amino-3,6-dioxy-sad, the amino piperidine carboxylic acid, 3-amino-propionic acid, amino Serine, amino tetrahydro pyran-4-carboxylic acid, arginine, l-asparagine, aspartic acid, azetidine (azetidine) carboxylic acid, the benzothiazolyl L-Ala, the butyl glycine, carnitine, the 4-chlorophenyl alanine, citrulline, Cyclohexylalanine, cyclohexyl statin (cyclohexylstatine), halfcystine, 2, the 4-DAB, 2, the 3-diaminopropionic acid, dopa, dimethylthiazole alkane carboxylic acid, L-glutamic acid, glutamine, glycine, Histidine, homoserine, oxyproline, Isoleucine, isonipecotic acid (isonipecotic acid), leucine, Methionin, methanoproline (methanoproline), methionine(Met), nor-leucine, norvaline, ornithine, the p-benzaminic acid, Trolovol, phenylalanine, phenylglycine, the piperidyl L-Ala, the piperidyl glycine, proline(Pro), the pyrrolidyl L-Ala, sarkosine, seleno-cysteine, Serine, statin (statine), the tetrahydropyrans glycine, thienyl alanine, Threonine, tryptophane, tyrosine, Xie Ansuan, alloisoleucine, allothreonine, 2,6-diamino-4-caproic acid, 2, the 6-diaminopimelic acid, 2, the 3-diaminopropionic acid, dicarboxidine, homoarginine, Homocitrulline, homocysteine, homocystine, hyperphenylalaninemia, high proline(Pro) and 4-hydrazino-benzoic acid.

Usually amino acid is divided into four groups with side chain character: tart, alkalescence, hydrophilic (polar) and hydrophobic (nonpolar).Amino acid described herein can identify by their full name or corresponding single-letter or trigram standard code.Use small letter single-letter code to show the D-chirality.

" amino acid analogue " refers to amino acid or amino acid whose small molecule mimetics, and it shares common chemistry, electric charge, space (steric) or other character with given amino acid.For example, the analogue of L-Ala comprises for example Beta-alanine, ethyl glycine, α-An Jiyidingsuan and D-L-Ala; The analogue of halfcystine comprises for example homocysteine, D-halfcystine and Trolovol; The analogue of phenylalanine comprises for example 3-fluorophenylalanine, 4-methylbenzene L-Ala, phenylglycine, 1-naphthyl L-Ala and 3,3-diphenylprop propylhomoserin, 4-amino-benzene L-Ala, penta fluoro benzene L-Ala, 2-pyridyl L-Ala, 3-pyridyl L-Ala, 4-oil of mirbane L-Ala, 2-pyrrolidyl phenylalanine, 3-piperidyl L-Ala, 4-piperidyl L-Ala; And the analogue of Histidine comprises for example 1-Methyl histidine, 2,4-DAB, thiazolyl L-Ala, 2,3-diaminopropionic acid, amidino groups L-Ala, 2-pyridyl L-Ala, 3-pyridyl L-Ala, 4-pyridyl L-Ala, thienyl alanine, ornithine, 4-Amidinophenylalaninederivatives and 4-amino-benzene L-Ala.

" amido protecting group " used herein refers to be used in any substituting group that prevents the amino experience chemical reaction on the molecule when chemical transformation occurs in other position in the molecule.The amido protecting group can be removed under suitable electrochemical conditions.The known many amido protecting groups of those skilled in the art, and the example of amido protecting group, their addition means and their method of removing can be referring to T.W.Green, Protective GroupsinOrganic Synthesis, John Wiley and Sons, New York, 1991; Chapter 7, M.Bodanszky, Principles of Peptide Synthesis, 1st and 2nd revised ed., Springer-Verlag, New York, N.Y. (1984) and (1993) and Stewart and Young, SolidPhase Peptide Synthesis, 2nd ed., Pierce Chemical Co., Rockford, III. (1984), the disclosure of quoting them here as a reference.Term " (mono-substituted) amino of protected property " refers to have the amido protecting group on (mono-substituted) amino nitrogen atom.In addition, term " shielded carbamyl " refers to have the amido protecting group on the carbamyl nitrogen-atoms.The example of this type of amido protecting group comprises formyl radical (" For "), trityl, phthalimido, the tribromo-acetyl base, chloracetyl, acetyl bromide and iodoacetyl, urethane (urethane) type capping group, such as tertbutyloxycarbonyl (" Boc "), 2-(4-xenyl) propyl group-2-oxygen carbonyl (" Bpoc "), 2-hydrocinnamyl-2-oxygen carbonyl (" Poc "), 2-(4-xenyl) isopropoxy carbonyl, 1,1-diphenyl-ethyl-1-oxygen carbonyl, 1,1-diphenyl propyl-1-oxygen carbonyl, 2-(3, the 5-dimethoxy phenyl) propyl group-2-oxygen carbonyl (" Ddz "), 2-(toluoyl base) propyl group-2-oxygen carbonyl, pentamethylene base oxygen carbonyl (cyclopentanyloxycarbonyl), 1-methylcyclopentane base oxygen carbonyl, cyclohexyl oxygen carbonyl, 1-methyl cyclohexane alkyl oxygen carbonyl, 2-methyl cyclohexane alkyl oxygen carbonyl, 2-(4-toluyl alkylsulfonyl)-ethoxycarbonyl, 2-(methyl sulphonyl) ethoxycarbonyl, 2-(triphenylphosphinyl)-ethoxycarbonyl, 9-fluorenylmethyloxycarbonyl (" Fmoc "), 2-(trimethyl silyl) ethoxycarbonyl, allyloxycarbonyl, 1-(trimethyl silyl methyl) third-1-thiazolinyl oxygen carbonyl, 5-benzisoxa oxalyl group methoxycarbonyl (5-benzisoxalylmethoxycarbonyl), 4-acetoxyl benzyl oxygen carbonyl, 2,2,2,-trichloro-ethoxycarbonyl, 2-ethynyl-third oxygen carbonyl, the cyclopropyl methoxycarbonyl, isobornyl oxygen carbonyl, piperidino oxygen carbonyl, benzyl oxygen carbonyl (" Cbz "), 4-phenylbenzyl oxygen carbonyl, 2-methyl-benzyl oxygen carbonyl, α-2,4,5-tetramethyl-benzyl oxygen carbonyl (" Tmz "), 4-methoxybenzyl oxygen carbonyl, 4-luorobenzyl oxygen carbonyl, 4-benzyl chloride base oxygen carbonyl, 3-benzyl chloride base oxygen carbonyl, 2-benzyl chloride base oxygen carbonyl, 2,4-dichloro benzyl oxygen carbonyl, 4-bromobenzyl oxygen carbonyl, 3-bromobenzyl oxygen carbonyl, 4-nitrobenzyl oxygen carbonyl, 4-cyanobenzyl oxygen carbonyl, 4-(oxygen base in the last of the ten Heavenly stems) benzyl oxygen carbonyl or the like; Phenacyl alkylsulfonyl, two thiophene succinyl (dithiasuccinoyl, " Dts "), 2-(nitro) benzenesulfinyl (" Nps "), xenyl-phosphine oxide group and similar amido protecting group.For example, the amido protecting group can be selected from Boc, Cbz and Fmoc.As long as the amino of derivatize is to subsequent reactions conditional stability and can remove and not destroy the rest part of compound at suitable point, just can adopt more than one amido protecting group.

Term used herein " aromatic series " refers to one, two or that other are a plurality of carbocyclic rings, aromatic nucleus system.Aromatic group can be chosen fusion wantonly to the one or more rings that are selected from aromatics, cycloalkyl, heterocyclic radical.Aromatics can have 5-14 ring members, such as for example 5-10 ring members.One or more hydrogen atoms can also be substituted base and replace, and described substituting group is selected from acyl group; amido; acyloxy; thiazolinyl; alkoxyl group; alkyl; alkynyl; amino; aromatic base; aryloxy; azido-; carbamyl; alkoxyl formyl (carboalkoxy); carboxyl; carboxy and amide groups (carboxyamido); carboxyamino (carboxyamino); cyano group; cycloalkyl; dibasic amino; formyloxy; guanidine radicals; halogen; heterocyclic base; heterocyclic radical; hydroxyl; imino-amino; mono-substituted amino; nitro; oxo; phosphine amino; sulfinyl; sulfoamino-; alkylsulfonyl; sulfo-; thio acylamino; thioureido and urea groups.The limiting examples of aromatic base comprises phenyl, naphthyl, indyl, xenyl and anthryl.

" die aromatischen Aminosaeuren " used herein refers to have the amino acid of the side chain that comprises aromatic ring structure.Die aromatischen Aminosaeuren comprises for example Histidine, tyrosine, tryptophane, phenylalanine, 1-naphthylalanine and 4-pyridyl L-Ala.

" carboxy protective group " refers to one of ester derivative of hydroxy-acid group, and it is usually used in sealing or protection hydroxy-acid group when other functional group on the compound is reacted.This type of carboxylic acid protective group's example comprises the tertiary butyl; the 4-nitrobenzyl; the 4-methoxybenzyl; 3; the 4-veratryl; 2; the 4-veratryl; 2; 4; 6-trimethoxy benzyl; 2; 4; the 6-trimethyl benzyl; the pentamethyl-benzyl; 3; the 4-Methylenedioxybenzyl; diphenyl-methyl; 4; 4 '-dimethoxytrityl; 4; 4 '; 4 "-trimethoxy trityl; the 2-phenyl propyl; trimethyl silyl; t-butyldimethylsilyl; benzoyl group; 2; 2,2-three chloroethyls; β-(trimethyl silyl) ethyl; β-(two (normal-butyl) methyl silicomethane base) ethyl; the p-toluenesulfonyl ethyl; 4-nitrobenzyl alkylsulfonyl ethyl; allyl group; cinnamyl; 1-(trimethyl silyl methyl)-propenyl and similar module (moiety).The kind of the carboxy protective group that is adopted is not crucial, as long as the carboxylic acid of derivatize is to subsequent reactions conditional stability and can remove and saboteur's rest part not at suitable point.The further example of these groups can be referring to E.Haslam, Protective Groups in OrganicChemistry, J.G.W.McOmie, Ed., Plenum Press, New York, N.Y. (1973), Chapter 5 and T.W.Greene, Protective Groups in Organic Synthesis, 2nd ed., JohnWiley and Sons, New York, N.Y. (1991), Chapter 5.Relational language is " a shielded carboxyl ", and it refers to the carboxyl with one or more above-mentioned carboxy protective group replacements.

" combination " and " combined " refers to the noncovalent interaction between polypeptide and another biologically active molecules, and its space and chemistry that depends on these two molecules is complementary.The space of polypeptide and chemistry are complementary by concrete aminoacid sequence decision.

" biologically active molecules " refers in biological environment (for example in organism, cell or their external model), can be by carrying out function or effect, perhaps peptide, nucleic acid and/or the small molecules of disease or illness treated in stimulation or response function, effect or reaction, such as the small molecules of organic or inorganic and their fragment.

" bridge " refers to the covalent linkage between other chemical module in two non-adjacent amino acid, amino acid analogue or the peptides.Bridge can be for example skeleton to skeleton, side chain to skeleton or side chain to the bridge of side chain.Bridge can prepare by the cyclization that causes new acid amides, ester, ether, thioether, alkene or disulfide linkage to form.Skeleton is for example formed due to the lactan by and C-end terminal at N-to the bridge of skeleton.

" cyclic peptide " refers to have the peptide of two non-adjacent amino acid whose intramolecular bonds of bridging.

Peptide used herein " dimer " refers to comprise the molecule of first and second peptide chains, and described first and second peptide chains can be identical or different.Dimer can further comprise at least one optional joint, and two peptides are covalently bond to this joint.

" dipeptide analogue " similar basically to dipeptides (for example first-class substantially row or have similar basically position or orientation) is such as for example glycylglycine.As long as identify structure qualification listed above, dipeptide analogue just can comprise any combination of the molecule of connection.Dipeptide analogue can have one or more peptide bonds, and its optional key that is selected from down group by means commonly known in the art replaces :-CH ₂NH-,-CH ₂S-,-CH ₂-CH ₂-,-CH=CH-(cis and trans) ,-COCH ₂-CH (OH) CH ₂-and-CH ₂SO-.The technician will recognize that dipeptide analogue also comprises for example β-corner stand-in.Referring to for example R.M.Friedinger, J.Med.Chem.46:5553-5566 (2003) and S.Hanessian, Tetrahedron 53:12789-12854 (1997).

The limiting examples of dipeptide analogue comprises:

(D, L-FriedingerShi lactan);

(L, L-FriedingerShi lactan);

-(3S)-3-amino-2-oxygen-1-piperidines-acetate and (3R)-3-amino-2-oxygen-1-piperidines-acetate;

-(3S)-3-amino-2-oxygen-1-azepine

(azepine) acetate and (3R)-3-amino-2-oxygen-1-azepine

Acetate;

-(3S)-3-amino-2-oxygen-1-pyrrolidine acetic acid and (3R)-3-amino-2-oxygen-1-pyrrolidine acetic acid;

-(3R)-3-amino-1-carboxymethyl-Valerolactim; And

-3-amino-N-1-carboxymethyl-2,3,4,5-tetrahydrochysene-1H-[1]-benzo-aza

-2-ketone.

Term " disulphide bridges " refers to the covalent linkage that forms between the sulfydryl (for example thiol group or sulfydryl) of 2 amino acid (such as for example halfcystine, Trolovol and homocysteine) after oxidation.Disulphide bridges can bridging be included in two amino acid in the same line peptide, causes the cyclisation of line peptide.Disulphide bridges also can form between two peptides, produces peptide dimer thus.

" structural domain " refers to have zone some distinctiveness physical features or effect, one or more peptide, for example the independent pleated sheet structure that is made of a section of peptide chain.Structural domain can be bonded to another identical or different structural domain.Structural domain can comprise the sequence of the distinctiveness physical features with peptide, and perhaps it can comprise the fragment with physical features, and this fragment has kept its binding characteristic (being that it can be bonded to second structural domain).

" effective dose ", " significant quantity ", " treatment significant quantity " or the like refer to that medicament is enough to provide physiology, pharmacology and/or the cognitive amount that changes of expectation, and it can change along with patient, disease and methods of treatment.Described amount can be to be used for the treatment of the dosage that it is believed that the experimenter who suffers from particular disorder, in this case, it should be enough to alleviate or improve the symptom of illness or illness, or preventive dose, in this case, it should be enough to partially or completely prevent the experimenter symptom to occur.

Term used herein " fusions " refers to the covalent coupling thing (conjugate) between peptide of the present invention and another molecule, and described another molecule can be for example protein, peptide, small molecules, polymkeric substance (for example polyoxyethylene glycol or polysaccharide) or nucleic acid.Peptide-small molecules or peptide-polymer fusions can prepare by synthetic, and peptide-protein or peptide-peptide fusions can prepare by chemical coupling or by expressing in appropriate host cell.

Term " regulation and control " refers to improve or reduce the level of bioactive peptide.Regulation and control can take place directly or indirectly.

Term used herein " polymer " refers to comprise the molecule of many peptide chains, and described peptide chain can be identical or different.Polymer can further comprise at least one optional joint, and at least two peptides are covalently bond to this joint.Polymer can be for example dimer, tripolymer or the tetramer.Joint can be for example two mercaptan joints, three mercaptan joints, four mercaptan joints, dicarboxylic acid joint, tricarboxylic acid joint, tetracarboxylic acid joint, amine joint, triamine joint or tetramine joint.

Term " peptide " refers to comprise with the amino acid whose molecule of the linear link coupled of amido linkage, and described amino acid comprises for example amino acid of L and D.Peptide can comprise amino acid derivative or non-amino acid module in addition, such as for example dipeptide analogue.The length range of peptide of the present invention can be 2-100,5-30,7-50, a 10-30 or 10-50 amino acid (comprising end value) for example, such as for example 11-35 amino acid.Peptide can comprise further modification, such as for example glycosylation, acetylize, phosphorylation, PEGization, lipidization (lipidation) or with the coupling of organic or inorganic molecule.

" positively charged " used herein refers to greater than 6 pH, such as the positively charged amino acid of for example pH6-8,6-9,7-8 or 7-9, amino acid analog thing or chemical module.For example, positively charged amino acid comprises Methionin, arginine and 2,4-diamino-butanoic.

Term " positively charged die aromatischen Aminosaeuren " refers to greater than 6 pH, such as for example positively charged amino acid of pH6-8,6-9,7-8 or 7-9.For example, positively charged die aromatischen Aminosaeuren comprises Histidine, 4-amino-benzene L-Ala and 4-Amidinophenylalaninederivatives.

With the aminoacid sequence of given sequence " substantially the same " can be for example at least 60%, 64%, 70%, 75%, 76%, 80%, 82%, 85%, 88%, 90%, 94%, 95%, 97%, 98% or 99% identical with given sequence.It can be by brachymemma, deletion, substitute or add at least one amino acid derived self-supporting sequencing row; And/or, can be because of for example adding, delete or substituting at least 1,2,3,4,5,6,7 or 8 amino acid and be different from given sequence.

" treatment " refers to reduce the seriousness or the time length of disease or illness; One or more symptoms that improvement is relevant with disease or illness; Beneficial effect is provided for the experimenter who suffers from disease or illness, and must cure this disease or illness; Perhaps preventing disease or illness.

Peptide " tripolymer " refers to comprise the molecule of first, second and the 3rd peptide chain, and described peptide chain can be identical or different.The peptide tripolymer can further comprise at least one optional joint, and at least two described peptides are covalently bond to described joint.

II. peptide of the present invention

Partly be bonded to the ability of FcRn based on Fc, peptide of the present invention assessed the relative affinity of FcRn and blocking-up IgG.Show the peptide of ability that is bonded to the Fc part of IgG in conjunction with FcRn and/or blocking-up FcRn and share some sequence general character or feature.So, one embodiment of the invention provide the Fc that can suppress human IgG partly to be bonded to the peptide of people Fc newborn infant acceptor, and it comprises sequence:

-Gly-X ₆-X ₇-X ₈-X ₉-X ₁₀-X ₁₁-

Wherein:

-X ₇Be selected from phenylalanine and phenylalanine analogues;

-X ₈With-X ₉Be selected from glycine, sarkosine, aspartic acid, D-amino acid, α-An Jiyidingsuan and their analogue independently of one another; Perhaps X ₈With X ₉One time-out forms dipeptide analogue;

-X ₁₁Be selected from tyrosine and tyrosine analogue; And

In certain embodiments, the length range of peptide is 7 to 50 amino acid and can be bonded to people FcRn, thereby prevents that FcRn is bonded to human IgG.

In an exemplary embodiment, peptide of the present invention comprises:

Gly-X ₆-Phe-X ₈-X ₉-X ₁₀-Tyr。

In another embodiment, peptide of the present invention can be represented with general formula:

R ₁-Gly-X ₆-X ₇-X ₈-X ₉-X ₁₀-X ₁₁-R ₂

Wherein:

-R ₁Has general formula X ₁-X ₂-X ₃-X ₄

Wherein

-X ₁Be selected from hydrogen, acyl group and amino acid blocking group;

-X ₂Lacking or be selected from amino acid and length is 2-15 amino acid whose peptide;

-X ₃The disappearance or can with X ₁₀, X ₁₂Or X ₁₃Form the amino acid of bridge, wherein said bridge be selected from N-terminal to the bridge of C-terminal, side chain to skeleton bridge and side chain to the bridge of side chain;

-X ₄Lacking or be selected from amino acid and length is 2-15 amino acid whose peptide;

-X ₆,-X ₇,-X ₈,-X ₉,-X ₁₀With-X ₁₁As above definition; With

-R ₂Has general formula-X ₁₂-X ₁₃-X ₁₄-X ₁₅

Wherein

-X ₁₂Disappearance or amino acid;

-X ₁₃Disappearance or amino acid;

-X ₁₄Lacking or be selected from amino acid and length is 2-15 amino acid whose peptide; With

-X ₁₅Be amino group or carboxy protective group.

When being used to limit peptide of the present invention, term " amino acid " comprises amino acid analogue.

In one embodiment, X ₆Be positively charged amino acid, be selected from Methionin, ornithine, 2,4-diamino-butanoic, 2,3-diaminopropionic acid, arginine, amidino groups L-Ala and their analogue.In another embodiment, X ₆Be die aromatischen Aminosaeuren, be selected from tyrosine, tryptophane, phenylalanine and their analogue.In another embodiment, X ₆Be positively charged die aromatischen Aminosaeuren, be selected from Histidine, 1-Methyl histidine, 2-pyridyl L-Ala, 3-pyridyl L-Ala, 4-pyridyl L-Ala, 4-amino-benzene L-Ala, 4-Amidinophenylalaninederivatives, thiazolyl L-Ala and their analogue.In another embodiment, X ₆It is the 4-Amidinophenylalaninederivatives.In another embodiment, X ₆Be selected from Histidine, 3-pyridyl L-Ala, 4-pyridyl L-Ala, 4-Amidinophenylalaninederivatives and their analogue.

Exemplary Histidine analogue is selected from but is not limited to: DAB, thiazolyl L-Ala, 2,3-diaminopropionic acid, amidino groups L-Ala, 2-pyridyl L-Ala, 3-pyridyl L-Ala, thienyl alanine, ornithine, 4-Amidinophenylalaninederivatives, 1-Methyl histidine, 4-amino-benzene L-Ala, 2-pyrrolidyl L-Ala, 3-piperidyl L-Ala and 4-piperidyl L-Ala.

Exemplary phenylalanine derivative can be selected from but be not limited to: 4-amino-benzene L-Ala; the penta fluoro benzene L-Ala; 2-pyridyl L-Ala; 3-pyridyl L-Ala; 4-oil of mirbane L-Ala; 1-naphthyl L-Ala; hyperphenylalaninemia; phenylglycine; 2-methylbenzene L-Ala; 3-methylbenzene L-Ala; 4-methylbenzene L-Ala; the 2-chlorophenylalanine; the 3-chlorophenylalanine; the 4-chlorophenylalanine; 3; 3-diphenylprop propylhomoserin; 4; 4-diphenylprop propylhomoserin; 4-tert.-butylbenzene L-Ala; Cyclohexylalanine; (4-glycyl)-phenylalanine; L-1; 2; 3,4-tetrahydroisoquinoline-3-carboxylic acid; D-Beta-methyl phenylalanine and L-Beta-methyl phenylalanine.

In one embodiment, X ₁₀Be selected from neutral and hydrophobic amino acid and their analogue.

In one embodiment, X ₂Being selected from amino acid and length is 2 or 3 amino acid whose peptides.In one embodiment, X ₂Comprise at least one hydrophobic amino acid.In another embodiment, X ₂Be that amino acid or length are 2-15 amino acid whose peptide, wherein C-terminal amino acid is hydrophobic amino acid.

In one embodiment, X ₄Be that amino acid or length are 2-15 amino acid whose peptide, wherein C-terminal amino acid is that N-is methylated.

In one embodiment, peptide is linear.In another embodiment, X ₁₀, X ₁₂Or X ₁₃In at least one be can with X ₃Form the amino acid of bridge, wherein said bridge be selected from N-terminal to the bridge of C-terminal, side chain to skeleton bridge and side chain to the bridge of side chain.In one embodiment, described bridge is the bridge of side chain to side chain, and it can be disulphide bridges, ether bridge, thioether bridge, alkene bridge or lactam bridge.

In one embodiment, described side chain to the bridge of side chain be following each between disulphide bridges: halfcystine and halfcystine; Halfcystine and homocysteine; Halfcystine and Trolovol; Homocysteine and homocysteine; Homocysteine and Trolovol; Or Trolovol and Trolovol.In another embodiment, described side chain to the bridge of side chain be following each between lactam bridge: aspartic acid and Methionin; Aspartic acid and ornithine; Aspartic acid and 2,4-diamino-butanoic; Aspartic acid and 2, the 3-diaminopropionic acid; L-glutamic acid and Methionin; L-glutamic acid and ornithine; L-glutamic acid and 2,4-diamino-butanoic; Or L-glutamic acid and 2, the 3-diaminopropionic acid.

In one embodiment, described peptide comprises at least one halfcystine (for example at X ₃) or cysteine analogs, described cysteine analogs is selected from homocysteine, D-halfcystine and Trolovol.

In one embodiment, X ₈And X ₉In at least one be D-amino acid or be selected from glycine, α-An Jiyidingsuan and sarkosine.

In one embodiment, X ₈With X ₉Form dipeptide analogue together, it is selected from:

-Beta-alanine;

-4-aminobutyric acid;

-5-aminovaleric acid;

-3-(aminomethyl) phenylformic acid;

-4-(aminomethyl) phenylformic acid;

-3-(aminophenyl) acetate;

-4-(aminophenyl) acetate;

-3-amino-2-oxygen-1-piperidines-acetate;

-(3S)-3-amino-2-oxygen-1-Piperidineacetic acid and (3R)-3-amino-2-oxygen-1-Piperidineacetic acid;

-(3S)-3-amino-2-oxygen-1-azepine

Acetate and (3R)-3-amino-2-oxo-1-azepine

Acetate;

-(3R)-3-amino-1-carboxymethyl-Valerolactim; With

-3-amino-N-1-carboxymethyl-2,3,4,5-tetrahydrochysene-1H-[1]-benzo-aza

-2-ketone.

In some embodiments; described peptide comprises at least one phenylalanine or phenylalanine analogues; it is selected from: tryptophane; tyrosine; 2-amino-benzene L-Ala; 3-amino-benzene L-Ala; 4-amino-benzene L-Ala; the penta fluoro benzene L-Ala; 2-pyridyl L-Ala; 3-pyridyl L-Ala; 4-oil of mirbane L-Ala; 1-naphthyl L-Ala; hyperphenylalaninemia; phenylglycine; 2-methylbenzene L-Ala; 3-methylbenzene L-Ala; 4-methylbenzene L-Ala; the 2-chlorophenylalanine; the 3-chlorophenylalanine; the 4-chlorophenylalanine; 3; 3-diphenylprop propylhomoserin; 4; 4 '-diphenylprop propylhomoserin; 4-tert.-butylbenzene L-Ala; Cyclohexylalanine; (4-glycyl) phenylalanine; L-1; 2; 3,4-tetrahydroisoquinoline-3-carboxylic acid; D-Beta-methyl phenylalanine; with L-Beta-methyl phenylalanine.

In some embodiments, described peptide comprises at least one tyrosine analogue, and it is selected from: phenylalanine, 4-amino-benzene L-Ala, 4-anisole L-Ala, penta fluoro benzene L-Ala, 2-pyridyl L-Ala, 3-pyridyl L-Ala, 4-pyridyl L-Ala, 4-oil of mirbane L-Ala, 2 nitrotyrosines and 4-fluorophenylalanine.

In one embodiment, X ₉With X ₁₀Form dipeptide analogue together, it is selected from:

-D, the L-FriedingerShi lactan

With

-L, the L-FriedingerShi lactan

In certain embodiments, peptide of the present invention comprises at least one Histidine analogue, it is selected from: 2,4-DAB, thiazolyl L-Ala, 2,3-diaminopropionic acid, amidino groups L-Ala, 2-pyridyl L-Ala, 3-pyridyl L-Ala, 4-pyridyl L-Ala, thienyl alanine, ornithine, Methionin, arginine, 4-Amidinophenylalaninederivatives, 1-Methyl histidine, 3-Methyl histidine, 1,3-dimethyl Histidine, 4-amino-benzene L-Ala, 2-pyrrolidyl L-Ala, 3-piperidyl L-Ala and 4-piperidyl L-Ala.

In certain embodiments, the amino acid no purpose scope between the amino acid of those formation bridges is 6-12.In other embodiments, between the amino acid that forms bridge, there are 8 or 9 amino acid.

In some embodiments, the length of peptide of the present invention is at least 7,50 amino acid of as many as.In other embodiments, the length of peptide is 11 to 35 amino acid.

In certain embodiments, peptide of the present invention exists with polymer, such as dimer, tripolymer or the tetramer.Polymeric each peptide can be identical or different.

In one embodiment, described peptide is a dimer, such as the dimer as the standard reductive alkylation product.In another embodiment, described dimer is the product that reacts between single peptide monomer and the multivalence joint.In one embodiment, described multivalence joint is selected from the joint of mercaptan, acid, alkohol and amine.In another embodiment, described dimer is the product of alkylated reaction, such as the alkylated reaction of for example mercaptan and alkylogen.

In one embodiment, this peptide is synthetic on resin, react multimerization (for example dimerization) with the multivalence joint then, the multivalence joint of described multivalence joint such as acid or amine is as described in embodiment 15.

In some embodiments, peptide of the present invention comprises the above-mentioned modification at least 2 places.

In certain embodiments, peptide of the present invention comprises:

A) be selected from down the aminoacid sequence of organizing:

QRFCTGHFGGLYPCNGP(SEQ?ID?NO：1)，

GGGCVTGHFGGIYCNYQ(SEQ?ID?NO：2)，

KIICSPGHFGGMYCQGK(SEQ?ID?NO：3)，

PSYCIEGHIDGIYCFNA (SEQ ID NO:4) and

NSFCRGRPGHFGGCYLF (SEQ ID NO:5); And

B) with SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 and SEQ IDNO:5 in the substantially the same aminoacid sequence of one or more sequences.

In some embodiments, described peptide comprises one or more sequences at least 64%, at least 70%, at least 76%, at least 82%, at least 88%, at least 94% or the 100% identical aminoacid sequence with SEQ ID NO:1, SEQ ID NO:2, SEQ IDNO:3, SEQ ID NO:4 and SEQ ID NO:5.In other embodiments, peptide of the present invention can comprise SEQ ID NO:1, SEQ ID NO:2, SEQ IDNO:3, SEQ ID NO:4 or SEQ ID NO:5 but have at least one place or the deletion of as many as 5 places, substitute or add sudden change.The all Toplink of the present invention suppress the combination of people FcRn to IgG.

In some embodiments, peptide of the present invention can comprise by at least one conserved amino acid and substitutes SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 or the SEQ IDNO:5 that modifies.In certain embodiments, peptide of the present invention can comprise SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 or the SEQ IDNO:5 that modifies by at least one amino acid replacement, wherein described amino acid is substituted with naturally occurring amino acid.In some embodiments, peptide of the present invention can have SEQ ID NO:1, SEQID NO:2, SEQ ID NO:3, SEQ ID NO:4 or the SEQ ID NO:5 that modifies by at least one amino acid replacement, wherein described amino acid is substituted with D-amino acid.In some embodiments, peptide of the present invention can have SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ IDNO:4 or the SEQ ID NO:5 that modifies by at least one amino acid replacement, wherein described amino acid is substituted with the N-amino acid that methylates.In some embodiments, peptide of the present invention can have SEQ IDNO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4 or the SEQ ID NO:5 that modifies by at least one amino acid replacement, wherein the amino acid replacement that described amino acid is existed with non-natural.In certain embodiments, peptide of the present invention can have SEQ ID NO:1, SEQ ID NO:2, SEQID NO:3, SEQ ID NO:4 or the SEQ ID NO:5 that modifies by at least one amino acid replacement, wherein described amino acid is substituted with the amino acid analog thing.

Perhaps, peptide of the present invention can comprise the aminoacid sequence that is selected from table 1, and it can be bonded to FcRn, suppresses the Fc part that FcRn is bonded to the IgG molecule thus.The present invention further comprises the peptide of the variant that comprises the listed sequence of table 1, and wherein said variant includes but not limited to: brachymemma; Share the peptide of at least 68%, 72%, 76%, 80%, 84%, 88%, 92% or 96% identity with at least a peptide in the table 1.Variant also comprises the peptide that comprises the listed sequence of table 1 but comprise at least one place amino acid replacement, and wherein said amino acid substitutes with amino acid, amino acid analogue or the amino acid analog thing that naturally occurring amino acid, non-natural exist.

Table 1

SEQ?ID?NO：6	AGQRFCTGHFGGLYPCNGPGTGGGK
SEQ?ID?NO：6	AGQRFCTGHFGGLYPCNGPGTGGGK	SEQ?ID?NO：7	AGGGCVTGHFGGIYCNTQGTGGGK
SEQ?ID?NO：8	AGKIICSPGHFGGMYCQGKGTGGGK	SEQ?ID?NO：7	AGGGCVTGHFGGIYCNTQGTGGGK
SEQ?ID?NO：8	AGKIICSPGHFGGMYCQGKGTGGGK	SEQ?ID?NO：9	AGPSYCIEGHIDGIYCFNAGTGGGK
SEQ?ID?NO：10	AGNSFCRGRPGHFGGCYLFGTGGGK	SEQ?ID?NO：9	AGPSYCIEGHIDGIYCFNAGTGGGK

In one embodiment, peptide of the present invention can comprise the listed aminoacid sequence of table 1 but have 1,2,3,4,5,6 or more a plurality of conserved amino acid substitute.In another embodiment, peptide of the present invention can comprise the listed aminoacid sequence of table 1 but

use

1,2,3,4,5,6 or more a plurality of naturally occurring amino acid replacement.

In some embodiments, peptide of the present invention can comprise the listed aminoacid sequence of table 1 but with 1,2,3,4,5,6 or the amino acid replacement that exists of more a plurality of non-natural.In another embodiment, peptide of the present invention can comprise the listed aminoacid sequence of table 1 but with 1,2,3,4,5,6 or more a plurality of N-amino acid replacement that methylates.In another embodiment, peptide of the present invention can comprise the listed aminoacid sequence of table 1 but

use

1,2,3,4,5,6 or more a plurality of D-form amino acid replacement.In another embodiment, peptide of the present invention can comprise the listed aminoacid sequence of table 1 but with 1,2,3,4,5,6 or more a plurality of amino acid analog thing substitute.

Exemplary embodiment of the present invention hereinafter is provided.Many these embodiments contain the listed aminoacid sequence of table 1 but are modified into and comprise one or more amino acid replacements.Though these embodiments provide suitable alternate example, other that should be appreciated that the present invention contains the biologic activity of not destroying peptide substitutes.

In an exemplary embodiment, peptide of the present invention comprises the aminoacid sequence of SEQ ID NO:6 but substitutes and the 12nd glycine substitutes to sarkosine and modify (wherein amino acid whose position is based on the amino acid numbering among the SEQ ID NO:6) to Trolovol with the 6th halfcystine.In another embodiment, peptide comprises the aminoacid sequence of SEQ ID NO:6 but modifies to alternative the substituting to the N-methylleucine with the 13rd leucine of Trolovol with the 6th halfcystine.

In another embodiment, peptide of the present invention can comprise SEQ ID NO:6 aminoacid sequence but with the 6th halfcystine to Trolovol substitute, the 11st and 12 two glycine branch be clipped to single (3R)-amino-1-carboxymethyl-2-Valerolactim substitute, and the 13rd leucine substitute to the N-methylleucine and modify.

In one embodiment, peptide of the present invention can comprise SEQ ID NO:6 aminoacid sequence but with the 6th halfcystine to Trolovol substitute, the 12nd glycine substitute, reach the 13rd leucine to sarkosine and substitute to the N-methylleucine and modify.In another embodiment, peptide of the present invention can comprise SEQ ID NO:6 aminoacid sequence but with the 6th halfcystine to Trolovol substitute, the 11st and 12 two glycine branch be clipped to single (3R)-amino-1-carboxymethyl hexanolactam substitute, and the 13rd leucine substitute to the N-methylleucine and modify.

In one embodiment, peptide of the present invention can comprise SEQ ID NO:6 aminoacid sequence but with the 6th halfcystine to Trolovol substitute, the 9th hyte propylhomoserin to 3-pyridyl L-Ala substitute, the 12nd glycine is alternative to sarkosine, reach the 13rd leucine substitutes to the N-methylleucine and modify.In another embodiment, peptide of the present invention can comprise SEQ ID NO:6 aminoacid sequence but with the 6th halfcystine to Trolovol substitute, the 9th hyte propylhomoserin to the 4-Amidinophenylalaninederivatives substitute, the 12nd glycine is alternative to sarkosine, reach the 13rd leucine substitutes to the N-methylleucine and modify.In another embodiment, peptide of the present invention can comprise SEQ ID NO:6 aminoacid sequence but with the 6th halfcystine to Trolovol substitute, the 9th hyte propylhomoserin to 4-pyridyl L-Ala substitute, the 12nd glycine is alternative to sarkosine, reach the 13rd leucine substitutes to the N-methylleucine and modify.

In certain embodiments, peptide of the present invention can exist with polymer, such as dimer, tripolymer or the tetramer.Polymeric each peptide can be identical or different.Each peptide can be as mentioned and is hereinafter carried out multimerization described in the embodiment.

In one embodiment, peptide of the present invention will be with K to the avidity of FcRn _DRepresentative, numerical range is that 50fM is to 1mM.In another embodiment, the avidity of peptide of the present invention will be with K _DRepresentative, numerical range is that 50fM is that 50fM is that 1pM is to 1nM to 1nM or numerical range to 100 μ M or numerical range.In another embodiment, the composition that comprises at least a peptide of the present invention will be regulated and control the serum-concentration of IgG.In one embodiment, peptide of the present invention can also be blocked the IgG constant region with interactional at least one amino acid of IgG constant region specificity among the FcRn and is bonded to FcRn by being bonded to.

III. generate the method for peptide of the present invention

1. the general method that synthesizes peptide of the present invention

Can use technology well known in the art to synthesize peptide of the present invention.For example, can use the polynucleotide of encoded peptide to be re-combined into peptide of the present invention in cell, it is whole to be made of naturally occurring amino acid.Referring to, Sambrook et al. for example, Molecular Cloning A Laboratory Manual, ColdSpring Harbor Laboratory, N.Y. (1989) and Ausubel et al., Current Protocols inMolecular Biology, Greene Publishing Associates and Wiley Interscience, N.Y. (1989).Perhaps, can use known synthetic method such as solid phase synthesis to synthesize peptide of the present invention.Synthetic technology is being known in the art.Referring to for example Merrifield, Chemical Polypeptides, Katsoyannis and Panayotis eds.pp.335-61 (1973); Merrifield, J.Am.Chem.Soc.85:2149 (1963); Davis et al., Biochem.Intl.10:394 (1985); Finn et al., TheProteins (3d ed.) 2:105 (1976); Erikson et al., TheProteins (3d ed.) 2:257 (1976).Can use standard Fmoc/tBu scheme, referring to W.C.Chan and P.D.White eds.Fmoc Solid Phase Peptide Synthesis:A Practical Approach Oxford UniversityPress Inc.New York (2000) and U.S. Patent No. 3,941,763.Perhaps, can be used in combination recombination method and synthetic method and synthesize chimeric protein of the present invention.In some applications, it can be useful using the combination of recombination method, synthetic method or reorganization and synthetic method.

2. the method that is used for the peptide analogs of synthetic peptide of the present invention

Conventional usage is followed in 20 kinds of conventional amino acid used herein and their abbreviation.Referring to Immunology--A Synthesis, 2 ^NdEdition, E.S.Golub and D.R.Gren, Eds., SinauerAssociates, Sunderland, Mass (1991).20 kinds of amino acid whose steric isomers of routine (for example D-amino acid); Alpha-non-natural amino acid is such as amino acid, the amino acid of α-two-replacement, N-alkyl amino acid, N-methylamino acid, the lactic acid of alpha-substitution; With other unconventional amino acid also can be the suitable component of polypeptide of the present invention.Unconventional amino acid whose example comprises: aminoisobutyric acid, 3-amino-1-carboxymethyl Valerolactim, 4-amidino groups-phenylalanine, 5-aminovaleric acid, 4-oxyproline, Gla, ε-N; N, N-trimethyl lysine, ε-N-ethanoyl Methionin, O-phosphoserine, N-ethanoyl Serine, N-formyloxy methionine(Met), 3-Methyl histidine, 5-oxylysine, σ-N-methylarginine, Trolovol, sarkosine and other similar amino acid and imino-acid (for example 4-oxyproline).According to the usage and the convention of standard, here in the polypeptide symbol of Shi Yonging, left-hand is to being aminoterminal direction and right-hand lay is the direction of C-terminal.

Conserved amino acid substitutes can contain the amino-acid residue that non-natural exists, typically synthetic rather than mix it by synthesizing in biosystem by chemical peptide.These comprise simulating peptide (peptidomimetic) and amino acid module other put upside down (reversed) or (inverted) form of inversion.

Based on common side chain character, naturally occurring residue can be classified:

1) hydrophobic: nor-leucine, Met, Ala, Val, Leu, Ile;

2) neutral hydrophilic: Cys, Ser, Thr, Asn, Gln;

3) tart: Asp, Glu;

4) alkalescence: His, Lys, Arg;

5) influence the residue of chain direction: Gly, Pro; With

6) aromatic: Trp, Tyr, Phe.

For example, the non-conservative alternative member that can involve a class in these classifications replaces with another kind of member.

Carrying out these when changing, according to some embodiment, hydrophilic index (hydropathic index) that can considered amino acid.Based on the hydrophobicity and the charge property of every seed amino acid, given the hydrophilic index assignment of every seed amino acid.They are: Isoleucine (+4.5), Xie Ansuan (+4.2), leucine (+3.8), phenylalanine (+2.8), halfcystine/Gelucystine (+2.5), methionine(Met) (+1.9), L-Ala (+1.8), glycine (0.4), Threonine (0.7), Serine (0.8), tryptophane (0.9), tyrosine (1.3), proline(Pro) (1.6), Histidine (3.2), L-glutamic acid (3.5), glutamine (3.5), aspartic acid (3.5), l-asparagine (3.5), Methionin (3.9), and arginine (4.5).

The importance of hydrophile amino acid number in giving protein interaction sexual biology function is understood in this area.Kyte et al., J.Mol.Bio.157:105-131 (1982). known can other has the amino acid of similar hydrophilic index or score value with some amino acid replacement, and still keep similar biologic activity.In the variation of carrying out based on hydrophilic index, in certain embodiments, comprise amino acid whose substitute of hydrophilic index within ± 2.In certain embodiments, be included in ± within 1 those, and in certain embodiments, be included in ± within 0.5 those.

This area is also understood and can be carried out similar amino acid whose substitute effectively based on hydrophobicity, particularly when intention is used to be bonded to specified protein target thing with the new functionalized protein of consequent biology or peptide.

Give these amino-acid residues with following wetting ability numerical value (hydrophilicity value): arginine (+3.0), Methionin (+3.0), aspartic acid (+3.0 ± 1), L-glutamic acid (+3.0 ± 1), Serine (+0.3), l-asparagine (+0.2), glutamine (+0.2), glycine (0), Threonine (0.4), proline(Pro) (0.5 ± 1), L-Ala (0.5), Histidine (0.5), halfcystine (1.0), methionine(Met) (1.3), Xie Ansuan (1.5), leucine (1.8), Isoleucine (1.8), tyrosine (2.3), phenylalanine (2.5), and tryptophane (3.4).When the variation of carrying out based on similar wetting ability numerical value, in certain embodiments, comprise amino acid whose substitute of wetting ability numerical value within ± 2, in certain embodiments, be included in ± within 1 those, and in certain embodiments, be included in ± within 0.5 those.

Use known technology, the technician can determine the suitable variant of the listed polypeptide of this paper.In certain embodiments, it is believed that to the unessential zone of activity that those skilled in the art can identify and can change in the molecule and do not destroy active appropriate area by target.In certain embodiments, the technician can identify residue and the module of guarding in the molecule between similar polypeptide.In certain embodiments, even to biologic activity or also can carry out conserved amino acid to the important zone of structure and substitute and do not destroy biologic activity or polypeptide structure is not had disadvantageous effect.

In addition, those skilled in the art can look back structure-functional study, thereby identify in the similar peptide activity or the important residue of structure.In view of such comparison, the technician can predicted polypeptide in in similar peptide to the importance of the important corresponding amino-acid residue of amino-acid residue of activity or structure.Those skilled in the art can select chemically similar amino acid to substitute and be predicted as important amino-acid residue like this.

Those skilled in the art can also analyze in the similar peptide three-dimensional structure and with the aminoacid sequence of the sort of structurally associated.Those skilled in the art can be created in each expectation amino-acid residue place and comprise single amino acid alternate test variant.Can use activation measurement well known by persons skilled in the art to screen variant then.These variants can be used to collect information about suitable variant.For example, if find the variation of particular amino acid residue cause active destroyed, desirably reduced or improper by non-, so just can avoid having the variant of this variation.In other words, based on the information of collecting from this normal experiment, those skilled in the art can easily determine such amino acid, and no matter should avoid there is individually or other sudden change ground of combination further substitutes.

According to some embodiment, amino acid replacement can be such: (1) has reduced proteoclastic susceptibility, (2) reduced susceptibility to oxidation, (3) changed the binding affinity relevant, and/or other physical chemistry or the functional performance of this peptide species have been given or modified in (4) with biological function.According to some embodiment, can in naturally occurring sequence, (in certain embodiments, in polypeptide, form intermolecular contacting structure territory part in addition) and carry out single or multiple amino acid replacement (in certain embodiments, conserved amino acid substitutes).In certain embodiments, typically, conserved amino acid substitutes not the constitutional features of material alterations parental array (for example, metathetical amino acid should not be tending towards interrupting being present in the spiral in the parental array, perhaps destroys distinctive other type secondary structure of parental array).The art-recognized polypeptide secondary or the example of tertiary structure be referring to Proteins, Structures andMolecular Principles, Creighton, Ed., W.H.Freeman and Company, New York (1984); Introduction to Protein Structure, C.Branden and J.Tooze, eds., GarlandPublishing, New York, N.Y. (1991); And Thornton et al., describe among the Nature 354:105 (1991).

In certain embodiments, amino acid derivative comprises covalent modification, includes but not limited to, with the chemical bonding of polymkeric substance, lipid or other organic or inorganic module.In certain embodiments, the specific-binding agent of chemically modified can have the longer circulating half-life of modifying than non-chemically of specific-binding agent.In certain embodiments, the specific-binding agent of chemically modified can have improved target ability at expectation cell, tissue and/or organ.In certain embodiments, with deutero-specific-binding agent covalent modification,, include but not limited to polyoxyethylene glycol, polyoxyethylene glycol or polypropylene glycol to comprise one or more water-soluble polymers appurtenants (attachment).Referring to for example US Patent No: 4,640,835; 4,496,689; 4,301,144; 4,670,417; 4,791,192; With 4,179,337.In certain embodiments, the deutero-specific-binding agent comprises one or more polymkeric substance, include but not limited to that mono methoxy-polyoxyethylene glycol, dextran, Mierocrystalline cellulose or other carbohydrate are polymer based, poly-(N-V-Pyrol RC)-polyoxyethylene glycol, propylene glycol homopolymer, poly(propylene oxide)/ethylene oxide copolymer, polyoxyethylene polyvalent alcohol (for example glycerine) and polyvinyl alcohol, and these mixture of polymers.

3. the method that is used for the analogue of synthetic peptide of the present invention

Peptide analogs usually is used as the non-peptide medicament with the character that is similar to the template peptide in pharmaceutical industry.The non-peptide compound of these types is called " peptide mimics " or " simulating peptide ".Fauchere, J.Adv.Drug Res.15:29 (1986); Veber and Freidinger, TINS is (1985) p.392; And Evans etal., J.Med.Chem.30:1229 (1987), this paper quotes it as a reference.Usually under the help of computerize molecule modeling, develop these compounds.Go up the structurally similar peptide mimics of useful peptide to treatment and can be used for producing similar treatment or preventive effect.Usually, simulating peptide is structurally similar such as people's antibody to example polypeptide (polypeptide that promptly has biochemical property or pharmaceutical activity), connects with the key metathetical peptide that is selected from down group but have one or more choosing wantonly by approach well known :-CH ₂NH-,-CH ₂S-,-CH ₂-CH ₂-,-CH=CH-(cis and trans) ,-COCH ₂-,-CH (OH) CH ₂-and-CH ₂SO-.In certain embodiments, can produce more stable peptide with one or more amino acid (for example D-Methionin displacement L-Methionin) that the D-amino acid system of same type substitutes consensus sequence.In addition, can produce the peptide that is tied (constrained peptides) (the Rizo and Gierasch comprise consensus sequence or substantially the same consensus sequence by means known in the art, Ann.Rev.Biochem.61:387 (1992), be incorporated herein by reference), such as the inside cysteine residues that for example can form the intramolecular disulfide bridge that makes the peptide cyclisation by interpolation.

Peptide dimerization or oligomerization can strengthen the avidity of peptide sequence to given acceptor.Referring to for example John, et al., Chem.Biol.12:939 (1997). can use several different methods well known in the art to synthesize the dimer of peptide of the present invention and the polymer of high-order more.Dimer or polymer can be as peptide sequence is directly synthetic continuously on the automated peptide synthesizer.Perhaps, can react to synthesize peptide multimer by each peptide monomer and multivalence joint module.Referring to for example Rose, J.Am.Chem.Soc.116:30 (1994). again for example, can synthesize peptide multimer by mixing ramose joint group before synthetic peptide sequence, (multiple antigenic peptides, synthesizing MAP) is the same as " multiple antigenic peptide ".D.Posnett et al., J.Biol.Chem.263:1719 (1988). the invention provides the novel method that is used to form these peptide dimers, comprise: in the time of on resin, the N-end of peptide is reacted such as for example succsinic acid with linkers, make the contiguous peptide on the solid-phase resin pearl react each other, form the terminal dimer that connects by peptide N-.Cutting to resin subsequently provides N-the terminal peptide dimer that connects.

4. make up the expression vector that is used to express peptide of the present invention

Can come the nucleic acid of composite coding peptide of the present invention by standard method known in the art, for example by using automatization dna synthesizer (such as can be) available from Biosearch, Applied Biosystems etc.Other nucleic acid synthetic method is known in this area.Referring to for example United States Patent(USP) Nos. 6,015,881; 6,281,331; 6,469,136.For recombinant production peptide of the present invention, the polynucleotide sequence of encoded peptide is inserted suitable expression, the carrier of transcribing and translate necessary element that promptly comprises the encoding sequence that inserts perhaps in the situation of rna virus vector, comprises and duplicates and translate necessary element.The nucleic acid of coding peptide of the present invention is inserted carrier, in correct reading frame.

The carrier that uses in the conversion contains the selection marker that is useful on the evaluation transformant usually.In bacterial system, this can comprise antibiotics resistance gene, such as penbritin or kantlex.The selection marker that uses in the mammalian cell of cultivating comprises the gene of giving drug resistance, such as Xin Meisu, Totomycin and methotrexate.Selection marker can be the selection marker that can increase.A kind of selection marker that increases is DHFR gene or DHFR cDNA.Simonsen and Levinson, Proc.Natl.Acad.Sci.USA80:2495 (1983) .Thilly (Mammalian Cell Technology, Butterworth Publishers, Stoneham, MA (1986)) summarized selection marker, and the selection of selection marker is fully within the ordinary skill level.Also selection marker and goal gene while transfered cell on the plasmid that separates perhaps can be imported them on same plasmid.If on same plasmid, selection marker and goal gene can be under the control of similar and different promotor, and a kind of arrangement in back produces bicistronic mRNA information.Such construction (for example U.S. Patent No. 4,713,339) is known in this area.

Expression element in the expression system they intensity and specificity aspect change.Depend on the host/vector system that is utilized, can in expression vector, use any suitable element of transcribing and translate, comprise composing type and epigamic promotor.For example, when in bacterial system, cloning, can use inducible promoter, such as pL, plac, ptrp, ptac (ptrp-lac hybrid promoter) or the like of phage; When in insect cell system, cloning, can use promotor such as baculovirus polyhedrin body promotor; When in the vegetable cell system, cloning, can use derived from vegetable cell genome (heat-shocked promotor for example; The promotor of RUBISCO small subunit; The protein-bonded promotor of chlorophyll a/b) or derived from plant virus (the 35S RNA promotor of CaMV for example; The coat protein promotor of TMV) promotor; When in mammal cell line system, cloning, can use derived from mammalian cell genome (for example metallothionein promoter) or derived from mammalian virus (gland virus stage starting for example; Vaccinia virus 7.5K promotor) promotor; When generation comprises the clone of expression product of multiple copied, can use carrier with the appropriate selection sign based on SV40, BPV and EBV.

In the situation of using plant expression vector, can drive the sequence expression of the chimeric protein of the present invention of coding linearity or non-cyclisation form by any promotor.For example, can use viral promotors, coat protein promotor (Takamatsu et al., EMBO is (1987) J.6:307-311) such as 35S RNA and 19S RNA promotor (Brisson et al., Nature 310:511-514 (1984)) or the TMV of CaMV; Perhaps, can use plant promoter, (Coruzziet al., EMBO be (1984) J.3:1671-1680 such as the promotor of RUBISCO small subunit; Broglie et al., Science 224:838-843 (1984)) or heat-shocked promotor, for example soybean hsp17.5-E or hsp17.3-B (Gurley et al., Mol.Cell.Biol.6:559-565 (1986)).Can use Ti-plasmids, Ri plasmid, plant viral vector, directly DNA conversion, microinjection, electroporation etc. that these constructions are imported vegetable cell.Weissbach ﹠amp; Weissbach, Methods for Plant Molecular Biology, Academic Press, NY, Section VIII, pp.421-463 (1988) and Grierson ﹠amp; Corey, Plant Molecular Biology, 2d Ed., Blackie, London, Ch.7-9 (1988).

Can be used for a kind of insect expression system of production chimeric protein of the present invention, autographa california (Autographa californica) nuclear polyhedrosis virus (AcNPV) is being expressed alien gene as carrier.This viral growth is in fall army worm (Spodoptera frugiperda) cell.Encoding sequence can be cloned as viral nonessential region (for example polyhedrosis gene), and place under the control of AcNPV promotor (for example polyhedron promotor).The successful insertion of encoding sequence will cause the deactivation of polyhedrosis gene and not close the recombinant virus of (non-occluded) generation of (promptly lacking the virus by polyhedrosis gene encoded protein matter character shell).Then these recombinant viruses are used to infect the fall army worm cell, wherein the gene that is inserted is expressed.Referring to for example Smith et al., J.Virol.46:584 (1983); U.S. Patent No. 4,215, the further example of 051. this expression system can be referring to Ausubel et al., eds., CurrentProtocols in Molecular Biology, Vol.2, Greene Publish.Assoc.﹠amp; Wileylnterscience (1989).

The another kind of system that can be used for expressing peptide of the present invention is the NADPH-linked glutamate synthase gene expression system, is also referred to as " GS expression system " (Lonza Biologies PLC, Berkshire UK).This expression system write up is in U.S. Patent No. 5,981,216.

In mammalian host cell, can utilize many expression systems based on virus., encoding sequence can be connected to adenovirus and transcribe/translate control complex body, for example late promoter and tripartite leader[as in the situation of expression vector in adenovirus.Can this mosaic gene be inserted the adenoviral gene group by reorganization in external or the body then.That be inserted in that (for example E1 or E3 district) in the virus genomic nonessential region will cause surviving and can be in infected host the recombinant virus of expression of peptides.Referring to for example Logan ﹠amp; Shenk, Proc.Natl.Acad.Sci.USA 81:3655 (1984).Also can use bovine vaccine 7.5K promotor.Referring to for example Mackett et al., Proc.Natl.Acad.Sci.USA 79:7415 (1982); Mackett et al., J.Virol.49:857 (1984); Panicali et al., Proc.Natl.Acad.Sci.USA79:4927 (1982).

5. in suitable target cell, express peptide of the present invention

Can go into suitable target cell and express polypeptide of the present invention expressing media transfection or cotransfection.Rotaring dyeing technology known in the art includes but not limited to calcium phosphate precipitation (Wigler et al., Cell14:725 (1978)), electroporation (Neumann et al., EMBO, J.1:841 (1982)) and based on the reagent of liposome.Can utilize multiple host-expression vector system to express chimeric protein described herein, comprise protokaryon and eukaryotic cell.These include but not limited to recombinant phage dna or plasmid DNA expression vector microorganism transformed through comprising suitable encoding sequence, such as bacterium (for example intestinal bacteria); Yeast or filamentous fungus that recombination yeast through comprising suitable encoding sequence or expressed in fungi carrier transform; The insect cell system that recombinant virus expression vector through comprising suitable encoding sequence (for example baculovirus) infects; Recombinant virus expression vector through comprising suitable encoding sequence (for example cauliflower mosaic virus or tobacco mosaic virus (TMV)) infects or the recombinant plasmid expression vector through comprising suitable encoding sequence (for example Ti-plasmids) plant transformed cell system; Perhaps the zooblast system comprises mammalian cell (for example CHO, Cos, HeLa cell).

6. be used for the synthetic method that comprises the fusion molecule of peptide of the present invention

In some embodiments, peptide of the present invention exists as fusion rotein, and described fusion rotein comprises peptide of the present invention and fusion partner.In one embodiment, described fusion partner gives characteristic for peptide of the present invention, such as one in the transformation period that prolongs, stability and the enhanced transhipment or multinomial, and/or can strengthen in the body or external effect.

In addition, can the each several part of the constant domain of heterologous polypeptide of the present invention and immunoglobulin (Ig) (IgG) is linked together, cause chimeric polyeptides.These specific fusion molecule make purifying demonstrate the transformation period of increase more easily and in vivo.This for example has been apparent in the chimeric protein of being made up of the various structural domains of preceding two structural domains of people CD4 polypeptide and mammalian immune sphaeroprotein heavy chain or constant region of light chain.EP 0 394 827; Traunecker et al., Nature, 331:84-86 (1988). in some cases, the fusion molecule that has the continuous dimeric structure of disulfide linkage owing to the IgG part can ratio such as independent monomer polypeptide or more effectively combination and other molecule of neutralization of polypeptide fragment.Referring to for example Fountoulakis et al., J.Biochem.270:3958-3964 (1995).

The present invention also provides the polynucleotide of the fusion molecule of the above-mentioned any peptide of encoding.This polynucleotide can be the parts of carrier, and described carrier also comprises the adjusting sequence that is used to transcribe polynucleotide.The polynucleotide of fusion molecule of the host cell that comprises any above-mentioned fusion molecule, coding any of these peptide or the carrier that the polynucleotide and being used to that comprise the fusion molecule of coding any of these peptide are transcribed the adjusting sequence of these polynucleotide have been the present invention further provides.The present invention also further provides composition, and it comprises any above-mentioned fusion molecule or Nucleotide, and/or any above-mentioned carrier or host cell and damping fluid or pharmaceutically acceptable carrier.

A. the fusions that comprises the antibody Fc structural domain

In one embodiment, the Fc structural domain coupling of peptide of the present invention and IgG can be increased its circulating half-life.In certain embodiments, peptide is covalently bound to the Fc structural domain.Reorganization as well known to those skilled in the art and semi-synthetic the method that produces the chimeric protein that comprises the Fc immunoglobulin domains.For example, aldehyde can be mixed peptide derivant, and use to the N-end of Fc optionally the standard reductive alkylation reaction come to react with Fc protein.Kinstler, Adv.Drug Del.Rev.54:477 (2002). perhaps, the peptide thioesters is reacted with the Fc that carries N-terminal cysteine residue.Dawson?and?Kent，Ann.Rev.Biochem.69：923(2000).

Owing to additionally added two FcRn binding sites by the Fc structural domain, this peptide-Fc merges derivative can have the interactional ability of blocking-up IgG-FcRn of increase.This peptide-Fc fusions also can be protected peptide to avoid degrading and so strengthen effect in the body of peptide.The fusion rotein that has the dimeric structure that disulfide linkage links to each other owing to IgG part can also be than peptide of the present invention more effectively combination and other molecule of neutralization, referring to for example Fountoulakis et al., and J.Biochem., 270:3958-3964 (1995).

At peptide of the present invention is to have in the embodiment of a fusion rotein part of IgG Fc structural domain, and people Ig Fc can comprise hinge, CH2 and the CH3 structural domain of human IgG such as human IgG1, IgG2 and IgG4.The present invention also provides the fusion molecule that comprises peptide of the present invention and variant Fc polypeptide or variant Fc polypeptide fragment, and wherein said variant Fc comprises hinge, CH2 and the CH3 structural domain of the human IgG2 with Pro331Ser sudden change, referring to U.S. Patent No. 6,900,292.

Comprise that with the suitable Fc structural domain of peptide link coupled of the present invention those select locations in the Fc district have the amino acid of sudden change to weaken its effector functions Fc structural domain of (comprising antibody dependent cellular cytotoxicity and CDC).For example, the Fc that has the Pro331Ser sudden change _{γ 2}Variant has than natural Fc _{γ 2}Little complement activity, and can not be bonded to Fc _γR.IgG4 Fc is defectiveness aspect the activating complement cascade, and it is to Fc _γThe binding affinity of R is than the low about order of magnitude of the most activated isotype IgG1.In one embodiment, peptide of the present invention is coupled to Fc with Leu235Ala sudden change _{γ 4}Variant is with natural Fc _{γ 4}Compare it and show minimum effector functions.In another embodiment, peptide of the present invention is coupled to have Leu234Val, the Fc of Leu235Ala and Pro331Ser sudden change _{γ 1}Variant, it also shows than natural Fc _{γ 1}Little effector functions.

B. comprise albuminised fusions

In certain embodiments, peptide of the present invention can be coupled to the white protein binding modules.This white protein binding modules-peptide conjugate can have the transformation period in the longer body, and so can require peptide dosage still less to realize the desired therapeutic effect.Chuang et al., Pharm.Res.19:569 (2002); U.S. Patent No. 6,685,179. so, one embodiment of the invention provide fusion molecule, it comprises peptide of the present invention and white protein fusion partner, and the latter comprises white protein, one or more white protein fragment, in conjunction with albuminised peptide and/or coupling lipid or can be in conjunction with the molecule of albuminised other molecule arranged.

The method that comprises albuminised fusion rotein that produces is known in this area.For example, adding peptide that cap reagent (capping reagent) modifies by hydrophobicity aromatic series has demonstrated non-covalent in conjunction with white protein and prolong the transformation period of peptide in rabbit.Zobel et al., Bioorg.Med.Chem.Lett.13:1513 (2003). again for example, demonstrated single free cysteine residues on the covalent attachment serum albumin with the peptide of thiol-reactive base group modification.Kim?et.al.，Diabetes?52：751(2003).

C. the fusions that has the PEGization module

In one embodiment, peptide of the present invention can PEGization single or multiple to comprise (for example 2-4) PEG module.Can implement PEGization by any PEGization reaction known in the art.The method that is used to prepare the PEGization protein generally comprises (a) and under some conditions polypeptide and polyoxyethylene glycol (such as reactive ester or the aldehyde derivatives of PEG) is reacted, and peptide of the present invention thus becomes and is attached to one or more PEG groups; And (b) obtain reaction product.Usually, based on the result of known parameter and expectation, be identified for the peak optimization reaction condition of these reactions with judging the case as it stands.

Those skilled in the art can utilize many PEG attachment meanss, referring to for example EP 0 401 384; Maliket al., Exp.Hematol., 20:1028-1035 (1992); Francis, Focus on Growth Factors, 3 (2): 4-10 (1992); EP 0 154 316; EP 0 401 384; WO 92/16221; And WO 95/34326.For example, the step that can carry out PEGization peptide of the present invention by acylation reaction or alkylation reaction with the reactive polyethylene glycol molecule.

So, peptide of the present invention comprises the peptide of PEGization, and wherein one or more PEG groups adhere to through acyl group or alkyl.This peptide can be single PEGization or many PEGization (for example those comprise the peptide of 2-6 or 2-5 PEG group).The PEG group generally is attached to peptide at amino acid whose α or the amino place of ε, but also contains any amino that the PEG group can be attached to peptide, and this amino is enough reactive so that be attached to the PEG group under proper reaction conditions.

Active ester derivative and peptide of the present invention that the PEGization of coming by acidylate generally comprises polyoxyethylene glycol (PEG) react.For acylation reaction, selected polymkeric substance typically has the single reaction ester group.Any reactive PEG molecule known or that find subsequently can be used to carry out the PEGization reaction.The example of suitable activatory PEG ester is that esterification is the PEG of N-hydroxy-succinamide (NHS).Acidylate used herein includes but not limited to peptide of the present invention and polymer being connected such as the following type between the PEG: acid amides, carbamate (carbamate), urethane (urethane) or the like.Referring to for example Chamow, Bioconjugate Chem., 5:133-140 (1994).Reaction conditions can be selected from the known or any reaction conditions of exploitation subsequently in PEGization field, but should avoid will deactivation peptide of the present invention condition, such as temperature, solvent and pH.

The PEGization of coming by acidylate generally will cause gathering the peptide of the present invention of PEGization.The key that connects can be an acid amides.Products therefrom can only have basically that (for example〉95%) is single, two or three PEGization.Yet, can on some amount, form the kind that some have higher degree PEGization, this depends on employed concrete reaction conditions.If desired, can isolate the more PEGization kind of purifying from mixture (particularly unreacted kind) by the standard purification technology, described standard purification technology comprises dialysis, saltouts, ultrafiltration, ion exchange chromatography, gel permeation chromatography and electrophoresis etc.

The PEGization of coming by alkylation is included in and exists reductive agent to make, and terminal aldehyde derivatives and the peptide of the present invention of PEG reacted.For the standard reductive alkylation reaction, selected polymkeric substance should have the single reaction aldehyde radical.Exemplary reactive PEG aldehyde is water miscible polyoxyethylene glycol propionic aldehyde or its single C1-C10 alkoxyl group or aryloxy derivative.Referring to for example U.S. Patent No. 5,252,714.

7. purifying is with the peptide of the present invention of biological method expression

Depend on employed expression system, improve the rules of setting up by this area then and separate expressed peptide of the present invention (for example affinity chromatography, size exclusion chromatography, ion exchange chromatography).

Expression vector can code tag, the chimeric protein that this label allows purification of Recombinant easily to produce.Example includes but not limited to carrier pUR278 (Ruther et al., EMBO is (1983) J.2:1791), and wherein the DNA with code book invention peptide connects into carrier, with lac z coding region in same reading frame, thereby produce hybrid protein; Can use the pGEX carrier to express to have the chimeric protein of the present invention of glutathione S-transferase (GST) label.These protein are generally solubility, and can be by to the absorption of gsh-sepharose 4B, then wash-out when having gsh, and purifying from cell easily.Carrier comprises and is used for cleavage site (zymoplasm or Xa factor proteolytic enzyme or the ProScission Protease that purifying is easily removed label afterwards ^TM(Pharmacia, Peapack, N.J)).

In order to increase production efficiency, the peptide of the present invention of a plurality of units that polynucleotide can be designed to encode is separated by the enzymatic cleavage site.Can cut gained polypeptide (for example handling) with recovering peptide unit by suitable enzyme.This can increase the output by the peptide of single promoters driven.When using in suitable virus expression systems, the inner translation of instructing by each peptide of the present invention of mRNA coding is IRES by internal ribosome entry site for example in transcript.As and, the polycistron construction instructs transcribing of single and big polycistronic mRNA, instructs the translation of a plurality of independent polypeptide then.This method has been eliminated the production and the enzymatic processing of polyprotein, and can significantly increase the output by the peptide of the present invention of single promoters driven.

The host cell that comprises the DNA construction of code book invention peptide can be cultivated in suitable growth medium, promptly be contained the required nutraceutical substratum of cell growth.The required nutrition of cell growth can comprise carbon source, nitrogenous source, indispensable amino acid, VITAMIN, mineral substance and somatomedin.Optional is that substratum can contain calf serum or foetal calf serum.In one embodiment, substratum is substantially free of IgG.Usually, growth medium will be selected the cell that contains the DNA construction, by for example medicament selection or by on the DNA construction or supply essential nutrition defective with the selection marker of DNA construction cotransfection.Usually, the mammalian cell of cultivating is contained cultivation in serum or the serum free medium (for example MEM, DMEM) in commercialization.Selection is that suitable substratum is within the ordinary skill level for employed specific cells.

Can in transgenic animal, produce peptide of the present invention,, mix alien gene in its genome such as rodent, ox, pig, sheep, goat or other non-human animals.Because this gene is present in the germ line tissue, it passes to the offspring from the parent.Foreign gene is imported unicellular embryo (Brinsteret al., Proc.Natl.Acad.Sci.USA 82:4438,1985).The method of generation transgenic animal known in the art comprises the transgenics of producing immunoglobulin molecules.Wagner?et?al.，Proc.Natl.Acad.Sci.USA?78：6376(1981)；McKnight?et?al.，Cell?34：335(1983)；Srinster?et?al.，Nature?306：332(1983)；Ritchie?et?al.，Nature?312：517(1984)；Baldassarre?et?al.，Theriogenology?59：831(2003)；Robl?et?al.，Theriogenology?59：107(2003)；Malassagne?et?al.，Xenotransplantation?10(3)：267(2003).

8. be used to screen and find in conjunction with FcRn and block the method for the interactional peptide of FcRn-IgG

Can use phage display library to identify peptide in conjunction with FcRn.Phage display library can easily produce, referring to Smith and Petrenko, and Chem.Rev.87:391 (1997).Perhaps, can obtain phage display library from commercial source, such as for example Dyax Corp. (Cambridge, MA).Depend on screening conditions, phage can be identified with various different propertiess.In order to identify peptide, can reach the combination of FcRn and the competition of IgG the phage library screening in conjunction with FcRn (and so combine) with IgG competition FcRn.Perhaps, can be by phage library is replaced acceptor (alternatereceptor) incubation with one or more so that from the library, eliminate in conjunction with the peptide that replaces acceptor.So, can from the phage set of expectation, cut out being bonded to the phage that replaces acceptor.By dna sequencing, can identify and in conjunction with FcRn and to suppress IgG-FcRn bonded peptide the phage clone that is bonded to FcRn.

Evaluation comprises in conjunction with the example of the additive method of the peptide of FcRn: mRNA shows (Roberts andSzostak, Proc.Nat.Acad.Sci.USA 94:12297 (1997)), based on displaying (the Boderand Wittrup of cell, Nat.Biotechnol.15:553 (1997)), reach synthetic peptide library (Lam, Nature354:82 (1991); Houghten et.al., Nature 354:84 (1991)).

9. be used to measure in conjunction with FcRn and block the method for the interactional peptide of IgG:FcRn

Many methods can be used to assess peptide or simulating peptide in conjunction with FcRn and block the interactional ability of FcRn:IgG.For example, surperficial plasmon resonance (SPR) be well known in the art be used to assess method in conjunction with situation (Biacore AB, Uppsala, Sweden).Make in this way, (FcRn or IgG) is fixed on the spr sensor chip with one of binding partners, and another kind of mating partner is passed through from chip, this chip monitoring gained signal.In identical experiment, will pass through from chip as interactional competitor peptide to be assessed between IgG and the FcRn.Any decline of signal can be interpreted as the measurement of interactional ability between this peptide blocking-up FcRn and the IgG.

The additive method of the interactional possibility of mensuration IgG:FcRn known in this field inhibitor peptides.The IgG competition assay that a kind of such method is 96 well plate format.In this illustration assay method, the soluble human FcRn on 96 orifice plates is exposed to IgG and test peptides.The remaining combined IgG that detects by anti-IgG antibody and standard ELISA colouring reagents provides the measurement of the interactional ability of peptide blocking-up FcRn-IgG.

Also can implement the ability of blocking-up IgG-FcRn bonded peptide on cell, described cell can be in the clone of its cell surface expression people FcRn with exploitation through the DNA transfection of coding people FcRn.Can use in conjunction with competition assay wherein IgG-FcRn bonded inhibitor peptides and fluorescently-labeled IgG molecule competition.Can measure the residue IgG level that is bonded to cell by for example standard fluorescence active cells letter sorting art (FACS).

10. the purposes of peptide of the present invention

Peptide of the present invention in conjunction with FcRn and the Fc part that suppresses the IgG constant region in conjunction with FcRn, thereby the IgG katabolism that causes IgG katabolism when not having peptide of the present invention increases.In exemplary embodiment, the IgG constant region is from IgG1, IgG2, IgG3 or IgG4 subclass.In specific embodiment, the IgG constant region is from IgG1, IgG2 or IgG4 subclass.Therefore, peptide of the present invention is useful for any desired catabolic disease of increase IgG of treatment or illness.For example, peptide of the present invention can be used for treating autoimmune disorder, inflammatory conditions, has etiologic etiological cardiovascular disorder (for example arteriosclerosis), transplant rejection and/or graft versus host disease (GVH disease) (GVHD) based on inflammation.Peptide of the present invention also can be used for detecting the FcRn in patient or the biological sample (for example body fluid, tissue or cell sample, cell culture supernatant liquid).Peptide of the present invention also can be used for purifying FcRn from biological sample (for example body fluid, tissue or cell sample, cell culture supernatant liquid).

So, the invention provides adjusting is the method for the state of an illness of feature with improper IgG amount or the level of expressing IgG antibody or non-expectation, comprises the peptide of the present invention of administering therapeutic significant quantity.In one embodiment, the described state of an illness is selected from inflammatory diseases, autoimmune disorder or cancer.In other embodiments, the invention provides the method for regulating the state of an illness by regulation and control IgG serum-concentration.In certain embodiments, method of the present invention can be used for treating, prevention or metering needle be to the immune response of therapeutic protein, stores up enzyme or thrombin such as for example Fibrinogen, thrombogen, factor V, factor VII, Factor IX, factors IX, factor X, factor XI, plasma thromboplastin antecedent, factor XI, plasma thromboplastin antecedent I, factor XI, plasma thromboplastin antecedent II or the von Willebrand factor such as for example erythropoietin, lysosome.In other embodiments, method of the present invention can be used for treating, prevention or metering needle be to the immune response of gene therapy vector.

A. autoimmune disorder

Peptide of the present invention can be used for treating autoimmune disorder, include but not limited to alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, (Autoimmune Addison ' sDisease) for the autoimmunity Addison's disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmunity lymphoproliferative syndrome (Autoimmune Lymphoproliferative Syndrome, ALPS), autoimmune thrombocytopenic purpura (ATP), Behcet (Behcet ' s Disease), bullous pemphigoid, myocardosis, sprue-dermatitis herpetiformis (Celiac Sprue-Dermatitis Herpetiformis), confirmed fatigue immune dysfunction syndromes (Chronic Fatigue Immune Dysfunction Syndrome, CFIDS), chronic inflammatory demyelination polyneuropathy (Chronic Inflammatory DemyelinatingPolyneuropathy), cicatricial pemphigoid, the CREST syndromes, cold agglutinin disease, Crohn disease (Crohn ' s Disease), the De Gesishi disease (Degos ' Disease), dermatomyositis, juvenile form dermatomyositis (Dermatomyositis-Juvenile), discoid lupus, the special property sent out mixed type cryoglobulinemia (EssentialMixed Cryoglobulinemia), fibromyalgia-fibromyositis, Graves' disease (Graves ' Disease), Ge-Ba Er Shi disease (Guillain-Barr é), struma lymphomatosa (Hashimoto ' sThyroiditis), idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (ITP), IgA nephropathy, insulin-dependent diabetes, juvenile arthritis, lichen planus, lupus, (M é niere ' sDisease) for Meniere, mixed connective tissue disease, multiple sclerosis, myasthenia gravis (MG), pemphigus, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndrome, polymyalgia rheumatica, PM-DM, primary agammaglobulinemia, primary biliary cirrhosis, psoriatic, Raynaud's phenomenon (Raynaud ' s Phenomenon), conjunctivo-urethro-synovial syndrome (Reiter ' sSyndrome), rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma, siogren's syndrome (

Syndrome), stiff man syndrome (Stiff-Man Syndrome), Takayasu arteritis (TakayasuArteritis), temporal arteritis/giant cell arteritis, transplant rejection, ulcerative colitis, uveitis, vasculitis, vitiligo and Wei Genashi granulomatosis (Wegener ' s Granulomatosis).

In one embodiment, described autoimmune disorder is selected from bullous pemphigoid, idiopathic thrombocytopenic purpura (ITP), myasthenia gravis (MG), pemphigus (for example pemphigus vulgaris) and transplant rejection.

In another embodiment, peptide of the present invention and steroid can be united and be used for immunosuppression.

B. inflammatory conditions

Peptide of the present invention can be used for treating inflammatory conditions, include but not limited to asthma, ulcerative colitis and inflammatory bowel syndrome, it comprises rhinallergosis/sinusitis paranasal sinusitis, skin allergic reaction (urticaria (urticaria/hives), angioedema, atopic dermatitis), food allergy, drug allergy, insectean metamorphosis reaction transformation reactions, mastocytosis, sacroiliitis, it comprises osteoarthritis, rheumatoid arthritis and spondyloarthropathy.

C. need proteinic disease of administering therapeutic or illness

Frequently, when suffering from proteinic disease of the administering therapeutic of needs or illness, the experimenter will form the antibody at therapeutic protein, and it stops therapeutic protein to realize the therapeutic purpose of its expection then.Thereby, can be with peptide of the present invention and therapeutic protein coupling with by reducing the benefit that the IgG level strengthens therapeutic protein; Wherein, IgG antibody reduces responsible to the bioavailability of therapeutic protein.

Thereby, embodiment provides adjusting, treatment or prevention to be derived from method at illness, disease or the illness of the immunne response of thrombin, comprise the disclosed herein any peptide of cell with the treatment significant quantity contacted that wherein said thrombin is selected from Fibrinogen, thrombogen, factor V, factor VII, Factor IX, factors IX, factor X, factor XI, plasma thromboplastin antecedent, factor XI, plasma thromboplastin antecedent I, factor XI, plasma thromboplastin antecedent II or the von Willebrand factor.This method can be used for regulating, treat or prevent to suffer among the patient of hemophilia A for example or hemophilia B the immunne response at thrombin.In one embodiment, peptide of the present invention has been blocked the Factor IX inhibitor.In another embodiment, among the patient that present method can be used for regulating, treatment or prevention suffer from simple erythroid aplasia (PRCA) at for example the immunity of therapeutic erythropoietin should.

D. the disease or the illness that need gene therapy

The obstacle that the successful implementation gene therapy is treated disease or illness also comprises the therapeutic protein of transgenes encoding and may be the formation that is used to send the antibody of this genetically modified carrier specificity.Thereby, peptide of the present invention can with the gene therapy coupling, with by reducing the benefit that the IgG level strengthens coded therapeutic protein.Reduce in the responsible situation in the bioavailability of IgG antibody to gene therapy carrier or coded therapeutic protein, these methods are particularly useful.The gene therapy carrier can be a virus vector for example, such as adenovirus and adeno associated virus.The disease that can use gene therapy to treat includes but not limited to cystic fibrosis, hemophilia, PRCA, muscular dystrophy or lysosomal storage disease, such as for example familial splenic anemia (Gaucher ' s disease) and Fa Bulishi disease (Fabry ' s disease).

The in-vivo imaging of e.FcRn and detection

Peptide of the present invention also can be used for detecting the assay method of FcRn.In some embodiments, described assay method is to detect peptide of the present invention and FcRn bonded binding assay.In these assay methods, can be with FcRn or peptide immobilization of the present invention, and another (FcRn or peptide of the present invention) passed through from immobilized binding partners.FcRn or peptide of the present invention can carry out detecting ground mark.Suitable marker comprises radio isotope, includes but not limited to ⁶⁴Cu, ⁶⁷Cu, ⁹⁰Y, ¹¹¹In, ¹²⁴I, ¹²⁵I, ¹³¹I, ¹³⁷Cs, ¹⁸⁶Re, ²¹¹At, ²¹²Bi, ²¹³Bi, ²²³Ra, ²⁴¹Am, ²⁴⁴Cm and 99mTc-MDP; Has the enzyme (for example luciferase, peroxidase, alkaline phosphatase, beta-galactosidase enzymes or the like) that can detect product; Fluorescent agent and fluorescent marker, for example fluorescein isothiocyanate, rhodamine, phycoerythrin, Phycocyanins, C-, allophycocyanin, Phthalyldicarboxaldehyde, fluorescamine; The metal of emitting fluorescence, for example ¹⁵²Eu or other are attached to the lanthanon of peptide of the present invention by metal-chelating group such as EDTA; Chemiluminescence compound, for example luminol,3-aminophthalic acid cyclic hydrazide (luminol), different luminol,3-aminophthalic acid cyclic hydrazide (isoluminol), theromatic acridinium ester, acridinium salt, imidazoles and barkite (esteror); And bioluminescent compound, for example fluorescein or aequorin (green fluorescent protein), specific binding molecules, for example magnetic particle, microsphere, nanometer ball, luminescent quantum dot nanocrystal (luminescent quantum dot nanocrystal) or the like.

Perhaps, can use specificity, comprise for example detectable label and subordinate phase antibody or reagent the energy amplifying signal in conjunction with right.For example, peptide of the present invention can be coupled to vitamin H, and the interpolation coupling there is the strepto-affinity element of horseradish peroxidase as subordinate phase reagent.It is another kind of suitable for right that digoxin and anti-digoxin provide.In other embodiments, can be with the subordinate phase antibody coupling to enzyme, such as peroxidase, with the substrate combination that colour-change takes place when having peroxidase.Can measure whether bonded exists between peptide of the present invention and the FcRn by the whole bag of tricks, comprise chemiluminescent detection and scintillation counting on the flow cytometry, microscopy, roentgenogpaphy, fluorometry, color developing detection, phosphorescent substance imaging, film of dissociated cell.These reagent known in this field and their using method.

Use for in-vivo diagnostic, can make by the peptide of the present invention of using q.s through mark particular organization or even the illness imaging of specific cells, described illness is to the be expressed as feature of small part with FcRn.

Clinical trial and used the metal ion of extremely multiple suitable in-vivo tissue imaging.In order to use the radio isotope imaging, expect following feature usually: (a) to patient's low radiation dose; (b) the high photon output of nuclear medicine rules is implemented in permission in short duration; (c) ability that produces with sufficient amount; (d) acceptable penalty; (e) use preparation simply; (f) need not subsequently the patient is isolated.Usually these characteristics explain are as follows: (a) to the radioactive exposure of the organ of most critical less than 5rad; (b) behind infusion, can obtain single image in a few hours; (c) radio isotope does not decay because of the emission particle; (d) isotropic substance can easily detect; (e) transformation period (the Lamb and Kramer that is lower than four days, " Commercial Production ofRadioisotopes for Nuclear Medicine ", In Radiotracers For Medical Applications, Vol.1, Rayudu (Ed.), CRC Press, Inc., Boca Raton, pp.17-62).In one embodiment, described metal is technetium-99m.

Thereby, the invention provides a kind of method of the experimenter's of acquisition images of interior regions, comprise the experimenter is used the composition of significant quantity and the scintigram picture that record comes available from the radioactive metal decay, described composition comprises at least a peptide of the present invention that contains radioactive metal.Similarly, the invention provides the method that strengthens experimenter's interior region mr (MR) image, comprise the experimenter used the composition of significant quantity and writes down the MR image of experimenter's interior region that described composition comprises at least a peptide of the present invention that contains paramagnetic metal.

Other method provided by the invention comprises the method for sound wave (sonographic) image that strengthens experimenter's interior region, comprise to the experimenter and use the composition of significant quantity and write down the audiogram picture of experimenter's interior region that described composition comprises at least a peptide of the present invention that contains metal.In this application, described metal can be any avirulent heavy metal ion.The method of the radioscopic image that strengthens experimenter's interior region also is provided, has comprised to the experimenter and use the peptide combinations that contains metal and write down the radioscopic image of experimenter's interior region.Can use radioactive, avirulent heavy metal ion.

Peptide of the present invention can be connected to sequestrant, is recorded in U.S. Patent No. 5,326 such as those, the sequestrant in 856.Then can be with peptide-inner complex complex body radio-labeling so that provide preparation to be used to relate to the diagnosis or the treatment of the disease or the illness of IgG horizontal adjustment.Peptide of the present invention also can be used for U.S. Patent No. 5,449, and the method that discloses in 761 is used for imaging or radiotherapy so that produce radiolabeled peptide.

F. dosed administration and treatment pattern

Peptide of the present invention can intravenously, subcutaneous, intramuscular, oral, hypogloeeis, suck, hypogloeeis, nose, rectum, vagina or use by the lung path.Peptide of the present invention can heeling-in in the biological polymer solid support or be connected to the biological polymer solid support and slowly be released into desired site to allow chimeric protein.

The dosage of peptide of the present invention will depend on disease to be treated or illness, disease or illness seriousness, experimenter (result who comprises their sex, age, body weight and expectation), and employedly specifically use the path and change.Dosage range can be 0.1-100,000 μ g/kg body weight.In one embodiment, the scope of dosed administration is 1-10,000 μ g/kg.In another embodiment, the scope of dosed administration is 10-1,000 μ g/kg.In another embodiment, the scope of dosed administration is 100-500 μ g/kg.Peptide of the present invention can continuous administration or is used with specified time interval.Can adopt the external test method to determine the optimal dose scope and/or the timetable of dispenser.Those skilled in the art can easily determine other effective dose by the routine test of establishing dose response curve, and the amount that for example improves or reduce the necessary peptide of the present invention of IgG level can calculate from vivo test.The technician will easily understand dosage level and can be used as the seriousness of specific compound, symptom and experimenter to the function of the susceptibility of side effect and change, and those skilled in the art can easily determine the preferred dose of given compound by multiple means.For example, in order to calculate the dosage of peptide of the present invention, depend on employed particular agent, those skilled in the art can use the information about the necessary amount of realization desired effects that is easy to obtain.

G. medicinal compositions

The invention still further relates to the medicinal compositions that comprises at least a peptide of the present invention and pharmaceutically acceptable carrier or vehicle.The example of suitable pharmaceutical carrier is referring to the Remington ' s PharmaceuticalSciences of E.W.Martin.The example of vehicle can comprise starch, glucose, lactose, sucrose, gelatin, Fructus Hordei Germinatus, rice, flour, chalk, silica gel, sodium stearate, glyceryl monostearate, talcum, sodium-chlor, skim-milk, glycerine, propylene, ethylene glycol, water, ethanol or the like.Composition can also contain pH buffer reagent and wetting agent or emulsifying agent.

For oral dispenser, medicinal compositions can adopt the tablet or the capsule form of ordinary method preparation.Composition can also be prepared into liquid, for example as syrup or suspension.Liquid can comprise suspension agent (for example sorbitol syrups, derivatived cellulose or hydrogenation edible-fat), emulsifying agent (Yelkin TTS or Sudan Gum-arabic), non-aqueous media (for example Prunus amygdalus oil, oily ester, ethanol or fractionated vegetables oil) and sanitas (for example methyl p-hydroxybenzoate or propyl ester or Sorbic Acid).Prepared product can also comprise seasonings, tinting material and sweeting agent.Perhaps, composition can be rendered as dried product, use for making up with water or other suitable medium.

For sucking and the hypogloeeis dispenser, composition can adopt according to the tablet of conventional scheme and lozenge form.

For sucking dispenser, compound used according to the invention is sent easily with aerosol spray form, pressurized package or atomizer (for example among PBSs) the middle injection of described aerosol from suitable propelling agent is housed, described propelling agent is Refrigerant 12, trichlorofluoromethane, Dichlorotetrafluoromethane, carbonic acid gas or other suitable gas for example.In the situation of pressurised aerosol, thereby can determine dose unit by the amount that provides valve to send metering.The capsule and the cartridge case (cartridge) of for example gelatin that uses in sucker or the insufflator can be mixed with the powdered mixture that this compound and suitable powder matrix such as lactose or starch are housed.

Medicinal compositions preparation can be used for the parenteral dispenser undertaken by injecting.The preparaton that is used to inject can present with unit dosage form, for example in ampoule or in the multi-dose container of the sanitas that interpolation is housed.Composition can be taked the form such as the emulsion among suspension, solution or oil or the hydrophily Jie, and contains formula agent (formulatory agent), such as suspension agent, stablizer and/or dispersion agent.Perhaps, activeconstituents can be a powder type, for the suitable medium structure of apirogen water for example.

Medicinal compositions can also be mixed with suppository or the enema,retention for the rectum dispenser, for example contains conventional suppository bases such as theobroma oil or other glyceryl ester.

Peptide of the present invention can be connected to sequestrant, such as U.S. Patent No. 5,326,856 those sequestrants of being put down in writing.Peptide-sequestrant complex body radio-labeling can be related to the disease of adjusting IgG level or the diagnosis or the treatment of illness to provide preparation to be used to then.Peptide of the present invention can also be used for U.S. Patent No. 5,449, and the method that discloses in 761 is used for radiotherapy to produce radiolabeled peptide.

H. purifying FcRn

Peptide of the present invention also can be used for purifying FcRn.In some embodiments, the peptide covalent attachment is extremely suitable chromatography substrate is to form effective FcRn separating medium.The solution that will contain FcRn passes through from chromatography substrate then, causes that FcRn is non-covalent to be bonded to immobilized binding partners.The solution that contains FcRn can be from biological sample, such as body fluid, tissue or cell sample, cell culture supernatant liquid.By clean immobilized peptide with appropriate solution: the FcRn complex body discharges FcRn with suitable elute soln from chromatography substrate then to remove impurity, thereby with the FcRn purifying.

Can use and well known to a person skilled in the art that the number of chemical method is attached to suitable chromatography substrate with peptide of the present invention.For example, peptide of the present invention can be attached to the matrix that comprises suitable reactive group, described reactive group such as mercaptan, amine, carboxylic acid, alcohol, aldehyde, haloalkyl, N-alkyl-maleimide, N-hydroxyl-succinimido ester, epoxide, azanol, hydrazides.

In other embodiments, peptide of the present invention can be modified to comprise non-covalent chemical module or the peptide sequence that is bonded to suitable chromatography substrate.For example, can come modified peptides with the vitamin H module, and can be with its non-covalent chromatography substrate that comprises affinity fibroin matter that is bonded to.Perhaps, can with through the peptide modified with FcRn solution incubation, then with the gained mixture from suitable chromatography substrate by to separate FcRn: the peptide complex body.

Peptide is used for the example of the similar applications of affinity purification can be referring to Kelley et al., " Developmentand Validation of an Affinity Chromatography Step Using a Peptide Ligand forcGMP Production of Factor VIII ", In Biotechnology and Bioengineering, Vol.87, No.3, Wiley Inter Scienc, 2004, pp.400-412 and U.S. Patent No. 6,197,526.

Embodiment

Embodiment is intended to the pure example as the present invention, therefore not will be understood that and limits the present invention by any way.Embodiment also describes and has described in detail all respects and the embodiment of foregoing invention.Embodiment is not intended to show that experiment hereinafter is all that implement or experiment is only arranged.Made great efforts accuracy (for example quantity, temperature etc.), but should consider some experimental errors and deviation to guarantee employed numerical value.Except as otherwise noted, mark is a mark by weight, and molecular weight is a molecular-weight average, and temperature is with a degree centigrade expression, and pressure is normal atmosphere or near normal atmosphere.

Embodiment 1: the expression of soluble human FcRn (shFcRn)

In Chinese hamster ovary (CHO) cell, use described in glutamine synthase expression system such as the document soluble human FcRn cDNA clone, expression and purifying.Referring to U.S. Patent No. 5,623,053.Terminator codon is placed on after the 274th amino acids of protein sequence of people FcRn, strides the film district to remove.

Embodiment 2: personnel selection FcRn transfection HEK293 cell

Use the SuperFect transfection reagent (Qiagen, Valencia, CA) according to scheme transfection human embryo kidney (HEK) (HEK) 293 cells of manufacturer recommendation (ATCC, Manassas, VA).With total length FcRn cDNA construction shown in Figure 1 (C.M.Story et al., J.Exp.Med.180:2377-2381 (1994); N.E.Simister etal., Eur.J.Immunol.26:1527-1531 (1996)) (Invitrogen, Carlsbad is CA) to produce FcRn expression vector FcRn:pCDNA6 at first to be cloned into pcDNA6 as plasmid vector.With same people β shown in Figure 1 ₂M cDNA construction at first is cloned into pcDNA3 (Invitrogen) as plasmid vector to produce people β ₂M expression vector β ₂M:pcDNA3 (D.Gussow et.al., J.Immunol.139:3132-3138 (1987)).

That day before the transfection, with the HEK293 cell with 0.5-2.5 * 10 ⁶The inoculation of cell/100mm ware and in cDMEM in 37 ℃ and 5%CO ₂Cultivated 16 hours.The component of cDMEM comprises: 1L DMEM (Invitrogen#11995-065); 10ml 1M HEPES, pH7.55; 10mlMEM amino acid solution (Invitrogen #11130-051); 10ml MEM non-essential amino acid solution (Invitrogen#11140-050); 10ml 100mM Sodium.alpha.-ketopropionate (Invitrogen #11360-070); 10ml penicillin streptomycin liquid (Invitrogen #15140-148); 10ml L-glutaminate solution (Invitrogen#25030-081); The 2 mercapto ethanol (Invitrogen #21985-023) of 1ml in 55mM Dulbecco phosphate buffered saline (PBS) (DPBS); 100ml heat-inactivated fetal bovine serum (FBS) (Invitrogen).On transfection same day, with 5 μ g FcRn:pCDNA6 constructions and 5 μ g β ₂M:pCDNA3DNA is added into 290 μ LDMEM (Invitrogen).Solution is mixed the several seconds and centrifugal.Then, 60 μ L SuperFect transfection reagents (Qiagen) are added into dna solution and vortex concussion 10 seconds.With DNA/SuperFect solution in room temperature incubation 5 to 10 minutes, during will be from celliferous ware sucking-off substratum and clean cell once with 4ml PBS.After 5 to 10 minutes, the complete growth medium of 3ml (cDMEM) is added into DNA/SuperFect solution at the DNA/SuperFect incubation, solution is mixed, and be added into cell in the 100mm ware immediately.

Use DNA/SuperFect solution in 37 ℃ and 5% CO in cell ₂Cultivated 2 to 3 hours.From cell, remove the substratum that contains DNA/SuperFect solution, clean cell three times, and fresh cDMEM is added into cell with PBS.Cultivate after 48 hours, assess nutrient solution to determine whether that FcRn/ β takes place by immunoblotting assay ₂The transient expression of m.In addition, the ratio of cell with 1:4 gone down to posterity into the cDMEM that contains 250 μ g/L Geneticins (Invitrogen) (as microbiotic) and 5 μ g/L blasticidins (to select stable transfection of blasticidin resistance).After microbiotic selected for 4 weeks, survivaling cell is inoculated into 96 hole tissue culturing plates with the density of 1 cells/well.Finally, select 12 clones, and with each clonal expansion, then by FcRn and β ₂The immunoblotting assay of m is checked expression.This Analysis and Identification expression FcRn and β ₂293 clones 11 of m are for having high expression level, and so are used for subsequently assay method.

Embodiment 3: to phage library screening FcRn-IgG inhibitor

(Cambridge, MA) Xu Ke filobactivirus display libraries has been identified and can have been suppressed IgG Fc part to FcRn bonded peptide to obtain Dyax Corp. by screening.More particularly, with following three library couplings; In screening, use TN-9-IV, TN10-X, TN-11-I and TN-12-I.As described in the Dyax scheme, when the library is coated with at expression in escherichia coli and with clone's dilution, reflect the independent survival phage sum that is included in each library by the transformant number of establishing for each library.The transformant number of TN-9-IV, TN10-X, TN-11-I and TN-12-I is respectively 3.2 x 10 ⁹, 2 x 10 ⁹, 2.7 x 10 ⁹With 1.4 x 10 ⁹The another kind of mode of mentioning the absolute number of the phage that can survive in the given volume is the plaque-forming unit (pfu) of each unit volume of narration.

A. the damping fluid that uses in the phage selection

Following damping fluid is used to screen the peptide in conjunction with FcRn.

1.NZCYM meat soup: 10g NZ amine-A; 5g sodium-chlor; 5g microbial culture yeast extract (Difco); The 1g casamino acids; 1g anhydrous slufuric acid magnesium dust; Described composition is dissolved in 800ml water, transfers to pH7.5 with 1N sodium hydroxide, water complements to the 1L cumulative volume then, and autoclaving 20 minutes.

2. binding buffer liquid (BB): PBS, pH6 add 10mM EDTA.

C.NZCYM-T:NZCYM meat soup adds 12.5 μ g/ml tsiklomitsins.

3.HBSS-E:HankShi the balanced salt aqueous solution (Invitrogen) adds 10mM EDTA (Invitrogen).

4.Min A salt: 10.5g K ₂HPO ₄(dipotassium hydrogen phosphate); 4.5g KH ₂PO ₄(potassium primary phosphate); 1.0g (NH ₄) ₂SO ₄(ammonium sulfate) and 0.5g Trisodium Citrate are dissolved in 1L water.

5.LB meat soup: 10g microbial culture Tryptones; 5g microbial culture yeast extract; 10g sodium-chlor is dissolved in 1L water and autoclaving 20 minutes.

6.CBS pH2:50mM Trisodium Citrate; 150mM sodium-chlor; With HCl damping fluid is transferred to pH2, and filtration sterilization.

7.LB agar: 30g microbial culture Tryptones; 15g microbial culture yeast extract; 30g sodium-chlor is dissolved in 3L water, and autoclaving 20 minutes.

8.LB soft agar: 20g microbial culture Tryptones; 10g microbial culture yeast extract; 20g sodium-chlor; 14g Bacto-agar, slowly heating but not boil and make them be dissolved in 2L water.

9.TE damping fluid: 10mM Tris, 1mM EDTA, pH7.

B. screening scheme: the first round

The titre of normal each library of about 100 random libraries according to them merged, this means: with 24 μ L TN9-IV (1.3 x 10 ¹⁰Pfu/ μ L), 12.5 μ L TN10-X (1.6 x 10 ¹⁰Pfu/ μ L), 225 μ LTN11-I (1.2 x 10 ⁹Pfu/ μ L) and 48.7 μ L TN12-I (2.9 x 10 ⁹Pfu/ μ L) with ice-cold 17% polyoxyethylene glycol (PEG) of 189 μ LPBS, 75 μ L (molecular-weight average: 8000Da, Sigma-Aldrich, St.Louis, MO) and 75 μ L 3M sodium-chlor mix, and incubation on ice 30 minutes.With HBSS-E 293 of one T75 culturing bottle is cloned the ratio partition of 11 cells (embodiment 2) with 1:3.Cell transfer to the 1ml Eppendorf tube, is cleaned once with cold binding buffer liquid, remove supernatant liquor then.With cell and phage on turner in 4 ℃ of incubations 1.5 hours.Behind the incubation, cell is cleaned 5 times with the ice-cold BB of 1ml, each after with 1400rpm centrifugal 2 minutes.(Calbiochem, San Diego CA) come the strong bonded phage of wash-out at the human IgG of BB dialysis by adding 66 μ M.With phage-IgG mixture and cell in 4 ℃ of incubations 1 hour.In centrifugation step (centrifugal 2 minutes of 1400rpm) afterwards, clean the cell precipitation thing for the first time with 200 μ l, 66 μ M IgG, centrifugal (centrifugal 2 minutes of 1400rpm) cleans last then with 100 μ l IgG.The scavenging solution of IgG is merged mutually with the elutriant of IgG, obtain 500 μ l final volume.As described below, with phage titration in the elutriant and amplification.

C. phage titre

Phage solution is diluted with 100 times of dilution step.Typically, in a continuous manner 2 μ l phage solutions are added into 198 μ l NZCYM meat soups to reach until 10 ^-10Extent of dilution.When the growth of XL1 Blue MRF ' Bacillus coli cells is in logarithmic phase and when reaching 0.5 optical density(OD), the phage of dilution is added into the culture of XL1 Blue MRF ' Bacillus coli cells at A600 (at the ultraviolet absorptivity of 600nm).With culture in room temperature incubation 10 minutes.Then, the 0.5ml cells infected is added into the top-layer agar (50/50 mixture of LB meat soup and LB agar) of the about 55 ℃ fusing of 3.5ml, and on the standard agar plate, launches, in 37 ℃ of overnight incubation.Calculate titre from the flat board that contains 30 to 300 plaques.For contain 50 plaques, the coating from 10 ^-8The flat board of phage diluent can carry out following calculating: 10 times of dilution x10 in 50 plaque/500 μ l cells infected x courses of infection ⁸Phage extent of dilution=10 ⁸Plaque-forming unit/μ l.

When be necessary to carry out subsequently phage E LISA and during sequencing analysis, pick out the independent agar bolt (agar plugs) that contains the phage plaque with autoclaved pasteur pipet (Pasteur pipets).The agar bolt is inserted 96 holes aseptic round bottom tissue culturing plates (Greiner), add 100 μ l TE to each hole wherein.Wash-out goes out phage from plaque, spends the night in 2 hours or 4 ℃ at 37 ℃ of need.

D. phage amplification

The culture of XL1 Blue MRF ' Bacillus coli cells is cultivated in NZCYM meat soup-T, reached 0.5 optical density(OD) until culture at A600 from 1/100 extent of dilution of saturated overnight culture.By with cell centrifugal 15 minutes with 3500rpm, then resuspended to 1/20 of original volume with MinA salt, thereby with cell concentration.Take turns the selection back with one and be diluted to the 1ml final volume with Min A salt, and be added into the spissated bacterial cultures of 1ml from the phage of cell wash-out.Incubation was added into 2ml2X NZCYM meat soup with phage-cell mixture after 15 minutes in 37 ℃ of water-baths, and coated the big NUNC flat board that NZCYM adds 50 μ g/ml penbritins is housed, until having done.With flat board in 37 ℃ of incubations 14 to 18 hours.When having 20mlPBS, soft the scraping of bacterium colony that will spend the night and form with spreading rod.The PBS that will contain bacterium and phage is collected in the centrifuge tube.When having 10ml PBS, the bacterium that remains on the flat board is scraped once more, and collect.Last 10ml PBS rinsing liquid is applied to flat board, and combines with material that all scrape.With bacterial cell precipitation, and pour limpid supernatant liquor into another centrifuge tube by centrifugal (3500rpm, 15 minutes), secondary clearing again, and finally pour out once more.Then, the 17%PEG+3M NaCl of 0.15mL volume is added into supernatant liquor, it is mixed being incorporated in 4 ℃ of preservations and spending the night.The phage that comes collecting precipitation by centrifugal (8500xg, 30 minutes), afterwards, abandoning supernatant.The phage throw out is resuspended in the PBS of small volume, makes it clarification by of short duration rotation, and with the 17%PEG+3M NaCl of 0.15 volume once more with its precipitation.Final phage throw out is resuspended in PBS, and titration then is for the selection of next round is prepared.

E. second take turns

Phage library dilution with amplification makes and has only 10 random library equivalents to dilute into 1ml binding buffer liquid.Closing damping fluid with cold junction cleans untransfected 293 cells of 1/3rd T75 culturing bottles once.The included deduction step of removing following phage (described phage expression can be bonded to the peptide of the cell of not expressing FcRn) from the library is following carries out twice, is about to phage with non-infected cells incubation 15 minutes.Reclaim supernatant liquor.Then, close damping fluid with cold junction 293 clones, 11 cells of 1/3rd T75 culturing bottles cleaned once, and with phage in turner in 4 ℃ of incubations 1.5 hours.Close damping fluid with the 1ml cold junction cell is cleaned also centrifugal (1400rpm changeed 2 minutes) five times, use 200 μ L66 μ M human IgGs (in binding buffer liquid, dialysing) then by phage-cell-IgG mixture was come the strong bonded phage of wash-out in 1 hour in 4 ℃ of incubations.After centrifugal (1400rpm changeed 2 minutes), collect supernatant liquor, and throw out is cleaned with 200 μ L, 66 μ M IgG, then clean with 100 μ L, 66 μ M IgG.As described in the phage titre and phage amplification part of mark hereinafter, to phage titration in the elutriant and amplification.

F. third round

Take turns describedly as mentioned about second, carry out this and take turns.When third round is finished,, and use phage E LISA to measure the IgG-FcRn inhibitor to the phage titration in the elutriant.

G. phage E LISA

Implement the following step, identifying by enzyme-linked immunosorbent assay (ELISA) can be in conjunction with the phage encoded peptide of FcRn.At first, prepare following solution:

Buffer A: PBS+0.1% tween+0.5% BSA

Buffer B: 100mM MES, pH5.5+150mM NaCl+0.1% tween

Damping fluid C:50mM MES, pH6.0+150mM NaCl+0.1% tween

To be used to increases shows 0.5 optical density(OD) that is cultured to A600 in conjunction with the XL1 blue MRF ' culture of Escherichia coli of the phage of FcRn ability from the 1:100 extent of dilution of overnight culture.Then, each phage plaque elutriant of 10 μ l of as above preparing is added into 30 μ l XL1 blueMRF ' Bacillus coli cells in each hole of 96 orifice plates, and in room temperature incubation 15 minutes.Then, the 130 μ l NZCYM meat soups that will contain 50 μ g/ml penbritins are added into each hole, then flat board are incubated overnight in 37 ℃.

Prepare microtiter plate (Pierce) the plain bag of strepto-affinity quilt, the BSA sealing,, then it is spent the night in 4 ℃ of bags with the 1mg/ml biotinylation soluble human FcRn in the buffer A (embodiment 4, the A joint) with 200 μ l/ hole buffer A rinsings.Discard the damping fluid that contains FcRn, and with damping fluid C with dull and stereotyped rinsing secondary.Then, 70 μ l buffer B are added into each dull and stereotyped hole, then add the bacterial cultures that 30 μ l contain phage., flat board is cleaned five times after 1 hour in room temperature with 200 μ l damping fluid C.Then, 100 μ l being contained coupling has the damping fluid C of 1:10000 diluent of the anti-M13 antibody (Amersham Pharmacia) of HRP to be added into each hole.With flat board in room temperature incubation 1 hour.Then, flat board is cleaned 9 times with damping fluid C, goes on foot TMB (KPL) with one and develop the color, stop with 2M sulfuric acid after 5-15 minute, and with Spectra Max Plus microplate reader (plate reader) (Molecular Devices) at the 450nm reading.

H. the pcr amplification of phage DNA

Use PCR Core System II test kit the wash-out among the TE to be increased to be used for order-checking from the phage of plaque according to the specification sheets (Promega) of manufacturers.Then, the phage of 5ml wash-out is added into reaction mixture, it contains every kind of dNTP of 200 μ M, 500nM primer 3PCRUP (5 '-CGGCGCAACTATCGGTATCAAGCTG-3 ') (SEQ ID NO:11), 500nM primer 3PCRDN (5 '-CATGTACCGTAACACTGAGTTTCGTC-3 ') (SEQ ID NO:12), 1xTaq dna polymerase buffer liquid (10x:500mM KCl, 100mM Tris-HCl, pH9.0 in the time of 25 ℃, 1% Triton X-100,15mM MgCl ₂), the Taq archaeal dna polymerase of 1.25 units.To be reflected at and carry out following program on the MJ Research PCT-200 thermal cycler: 94 ℃ 5 minutes; 30 circulations, its consist of 94 ℃ 15 seconds, 55 ℃ of 30 seconds and 72 ℃ 60 seconds; Then 72 ℃ 7 minutes.Use QiaQuick PCRPrep test kit (Qiagen) to come the purifying products therefrom according to the specification sheets of manufacturers, come quantitatively by the A260 absorbancy, and use primer 3SEQ-80 (5 '-GATAAACCGATACAATTAAAGGCTCC-3 ') (SEQ ID NO:13) to check order.

The dna sequence dna of the aminoacid sequence that provides in the code pattern 1 has been provided the sequencing result of the phage of amplification after the three-wheel screening.These " phage is hit thing (hits) " are used to identify the total polypeptide sequence jointly, and it defines with following aminoacid sequence: G-H-F-G-G-X-Y (SEQ ID NO:14).

Embodiment 4: peptide-IgG competitive ELISA

In order to determine that whether the peptide of the present invention that obtains derived from screening filobactivirus display libraries can also be blocked IgG and be bonded to FcRn, designs and implement following ELISA assay method.

A. biotinylation shFcRn

With the solution dialysis twice of soluble human FcRn (shFcRn) in the Tris damping fluid, in 2LPBS (pH8.0) 3 hours at every turn.Measure the quantity of the shFcRn of recovery by measuring the 280nm absorbancy.Optical extinction coefficient (ε=85880M by absorbancy reading and shFcRn ^-1Cm ^-1) multiplying each other obtains the concentration of shFcRn.(Invitrogen, Carlsbad CA) handle the biotinylation of finishing shFcRn in 2 hours to the shFcRn after the dialysis in 4 ℃ by the Sulfo-NHS-LC-vitamin H with 2 times of molar excess.Then, shFcRn-Sulfo-NHS-LC-biotin reaction mixture is dialysed twice in the cold PBS of 2L, then another time absorbancy reading is to measure remaining protein concentration.Biotinylated shFcRn is stored in 4 ℃ with 0.1% sodiumazide, until needs.

B. peptide-IgG competitive ELISA assay method

(IL) flat board of 96 hole ReactiBindNeutravidin of sealing bag quilt cleans twice for Pierce, Rockford with BSA with 200 μ l/ hole buffer A (buffer A: PBS pH7.4 (Gibco, 14040), 0.5% BSA does not have IgG, 0.05% tween 20).Quilt is wrapped with the biotinylation shFcRn of 100 μ l/ holes, 1 μ g/ml in buffer A in each hole.With flat board sealing, then in 37 ℃ of incubations 2 hours.Then, (buffer B: 100mM MES pH6,150mM NaCl, 0.5% BSA do not have IgG (Jackson Immuno Research, West Grove, PA), 0.05% tween 20) and clean flat board with 200 μ l/ hole buffer B.Then, (Calbiochem, San Diego CA) and the 50 various peptide competitors in μ l/ hole (different concns), make that the IgG final concentration in each hole is 3nM to add the human IgG of 50 μ l/ hole 6nM in buffer B.In order to make its mixing, flat board was shaken 2 minutes, sealing, and in 37 ℃ of incubations 2 hours.Behind the incubation, with liquid sucking-off from flat board, and the coupling of adding the 1:10000 dilution of 100 μ l/ holes in buffer B has the anti-human IgG F of goat (ab ') the fragments specific F (ab ') of peroxidase ₂Fragment (Jackson ImmunoResearch, West Grove, PA).With dull and stereotyped cover lid,, and clean 4 times with the ice-cold buffer B in 200 μ l/ holes in room temperature incubation 30 minutes.(KPL, Gaithersburg MD), and make flat board manifest until color in the room temperature incubation, and this will spend 5 to 10 minutes to add 100 μ l/ hole SureBlue tmb substrate solution.In case color manifests, (KPL, Gaithersburg MD), and measure the 450nm absorbancy to add 100 μ l/ hole TMP stop baths.With data with absorbancy to the peptide concentration mode 50% inhibition concentration (IC that derives that maps ₅₀) value.

Embodiment 5: peptide-IgG competition FACS assay method

Determine that except using the ELISA method of describing among the embodiment 4 whether the peptide of the present invention that obtains derived from screening the filobactivirus display libraries can also be blocked IgG and be bonded to FcRn on the cell, designs and implemented following fluorescence activated cell sorting (FACS) assay method.

A. use the Alexa-Fluor-488 mark

With Alexa Fluor 488 protein labeling test kits (Molecular Probes/Invitrogen, Carlsbad, CA) according to the scheme mark humanization IgG1 of manufacturers suggestion (

, Medlmmune, Gaithersburg, MD).In brief, 50 μ l 1M sodium bicarbonates (pH9.0) are added into the IgG solution of 500 μ l 2mg/ml in PBS.This protein soln is added into Alexa Fluor 488 succinimido esters (dry powder), then in room temperature incubation 1 hour.Use test kit composition post (Bio-RadBioGel P-30 Fine size exclusion purifying resin) to come protein purification by size exclusion chromatography.Sample is loaded on the post, and uses the PBS wash-out.Article one, painted band contains underlined protein.Measure the mark degree by 280nm and the 494nm absorbancy of measuring the wash-out IgG of institute.Use following formula to determine the protein volumetric molar concentration: protein concn (M)=[A ₂₈₀-(A ₄₉₄X 0.11) the x dilution factor]/203,000.In addition, being used for the proteinic formula of derivation mole dyestuff/mole is: A ₄₉₄X dilution factor/71,000x protein concn (M).Typically, every mole of IgG is mixed with 4-7 mole Alexa-Fluor 488.

B. the IgG-peptide competition FACS assay method of using 293 clones, 11 cells to carry out

When preparing for assay method, to contain 5 μ g/ml blasticidins and 250 μ g/ml G418 (Gibco, Carlsbad, CA) DMEM perfect medium (CA) sink for Gibco, Carlsbad by HEK 293 clone's 11 cells (embodiment 2) rotations in, and be resuspended in damping fluid C (damping fluid C: the DulbeccoShi PBS (Gibco that contains 10mM EDTA (Gibco), Carlsbad, CA)), concentration is 3 x 10 ⁶Cell/ml.With transfer pipet cell (0.1ml) is transferred to each holes of 96 hole assay plate, and use Sorvall RT7 desk centrifuge with 2600RPM with centrifugal 5 minutes of flat board.Supernatant liquor is poured out light and slowly and on paper handkerchief, flat board is blotted.To be added into flat board with the peptide competitor (90 μ l) that different concns is dissolved in damping fluid C, and mix with the hyperchannel micropipet.With 10 μ l Alexa, 488 marks Be added into each dull and stereotyped hole, make Alexa 488 marks Final concentration be 100nM.Flat board is wrapped in the tinsel, placed on ice 1 hour, in Sorvall RT7 desk centrifuge centrifugal 5 minutes subsequently with 2600rpm, then clean once with 100 μ l damping fluid C, and the second time centrifugation step.Cell is resuspended in 200 μ l damping fluid C, and on Beckman Coulter EPICS XL flow cytometer, analyzes.

Embodiment 6: use surperficial plasmon resonance (SRP) to measure the equilibrium association constant (K of peptide _D) method

Implement the following step to pass through as Biacore (BIAapplications Handbook, version AB, section 4.2, Biacore AB, Uppsala, the amine linked reaction of Sweden) being recommended is come crosslinked soluble human or macaque FcRn to CM5 sensor chip (Biacore AB, Uppsala, Sweden) dextran surface, described amine linked reaction involve 1-ethyl-(3-dimethylaminopropyl)-carbodiimide (EDC) (Biacore AB, Uppsala, Sweden) and N-hydroxy-succinamide (NHS) (Biacore AB, Uppsala, Sweden).(B iacore AB, Uppsala Sweden) are diluted to FcRn protein the concentration of 10 to 30 μ g/ml, and are used to wrap a flow cell by on the sensor chip with 50mM sodium acetate pH4.5.(Biacore AB, Uppsala Sweden) seal remaining position on the FcRn flow cell with 1M ethanolamine hydrochloric salt (pH8.5).To contrast the flow cell sealing to be used for thanomin with reference to deduction.In order to analyze monomeric peptide, with the whole density of FcRn bag quilt to the 4000-5000 unit of replying (RU).In order to analyze peptide dimer, with the density of FcRn bag quilt to 2000-2500RU.(BiacoreAB, Uppsala Sweden) carry out all SPR and measure to use BIACORE3000 equipment.In order to carry out the measurement of pH6 or pH7.4, will test that (Biacore AB, Uppsala carry out in Sweden) at 50mM phosphoric acid salt, 100mM sodium-chlor, 0.01% tensio-active agent P20.

A. be used to measure the representative rules of the binding constant of monomeric peptide

The peptide of ten 2 times of dilutions was injected 2 minutes on the FcRn-CM5 chip with the speed of 20 μ l/min.With damping fluid peptide was dissociated 2.5 minutes from chip.(Biacore AB, Uppsala Sweden) came to remove any remaining peptide from chip in 30 seconds to inject the HBS-P damping fluid with the speed of 30 μ l/min.Produce sensing figure, and (Biacore AB, Uppsala analyzes Sweden) to use 3.1 editions BioEval softwares.The balance RU that per injection is observed maps to concentration.The affine model of steady state that use is included in the BioEval software is analyzed described figure, thereby derives balance K _DValue.

B. be used to measure the representative rules of the binding constant of dimer peptide

The peptide of ten 2 times of dilutions was injected 10 minutes on the FcRn-CM5 chip with the speed of 30 μ l/min.With damping fluid peptide was dissociated 10 minutes from chip.The solution that contains 50mMTris-hydrochloric acid, 100mM NaCl, 0.01% tensio-active agent P20pH9.0 with 100 μ l/min injection in twice 60 seconds to remove any remaining peptide from chip.

Produce sensing figure, and (Biacore AB, Uppsala analyzes Sweden) to use 3.1 editions BioEval softwares.The balance RU that per injection is observed maps to concentration.The affine model of steady state that use is included in the BioEval software is analyzed described figure, thereby derives balance K _DValue.

Embodiment 7: monomeric peptide synthetic that contains disulfide linkage

The synthetic use solid-phase peptide of monomeric peptide is synthetic carries out, or with the agglomerating round-bottomed flask manually or by use Advanced Chemtech 396-omega synthesizer (Advanced Chemtech, Louisville, KY).The Fmoc/tBu scheme of use standard (W.C.Chan and P.D.White eds., Fmoc Solid Phase Peptide Synthesis:A Practical Approach Oxford UniversityPress Inc.New York (2000)), with Rink amide resins (Novabiochem, San Diego, CA) or PAL-PEG-PS (Applied Biosystems, Foster City, CA) coupling is to produce the C-terminal amide after cutting.Coupling agent is 2-(1H-benzotriazole-1-yl)-1,1,3, and 3-tetramethyl-urea phosphofluoric acid ester (HBTU) and N-hydroxybenzotriazole (HOBt) (Novabiochem, San Diego, CA).Alkali be diisopropylethylamine (DIEA) (MO), and solvent is N for Sigma-Aldrich, St.Louis, dinethylformamide (DMF) (EM Science, Kansas City, MO).Typical synthesis cycle comprises 10 minutes deprotection steps of 2 x that 20% piperidines that is used among the DMF carries out, the 2 x amino acid coupling of carrying out with HOBt/HBTU in 30 minutes, and 10 minutes of carrying out with diacetyl oxide/HOBt add the cap step.By 95% trifluoroacetic acid, 2.5%thanedithiol, 1.5% tri isopropyl silane and 1% water treatment 2 hours, peptide is cut down from resin, use ice-cold ether sedimentation then, centrifugal, and grind three times with ether.

(EM Science, Kansas City MO) are dissolved to peptide the concentration of 1mg/ml, thereby the crude product peptide that will contain halfcystine is oxidized to their corresponding disulphide by the mixture with acetate and water 4:1.With the iodine (the 1M solution in water) of 10 molar equivalents (Sigma-Aldrich, St.Louis MO) are added into solution, and with reaction mixture in mixed at room temperature 1 hour.Add gradually 1M Sulfothiorine (Sigma-Aldrich, St.Louis, MO) until obtaining clear soln, thus termination reaction.Reaction mixture is concentrated in a vacuum, and (Millford MA) comes purifying to use the Waters Prep600 reverse-phase HPLC system that is equipped with 250mm x 21.2mm Phenomenex (Torrence CA) C18 post subsequently.The elutriant of selecting for the HPLC purification step is the acetonitrile gradient that contains in the water of 0.1% (w/v) TFA.Suitable fraction is collected, merge, and freeze-drying.By (CA) link coupled makes up 250mm x 2mm post for Applied Biosystems, Foster City, and (anti-phase analysis HPLC CA) confirms the identity and the purity of peptide for Phenomenex, Torrence with electrospray ionization mass spectrum (Mariner ES-MS).

Table 2 provides the tabulation of initial phage display peptide sequence, and they are used to identify to have the high-affinity of people FcRn and the peptide of the interactional ability of blocking-up IgG-FcRn derived from the screening of peptide expression library.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

Table 2: initial phage display peptide sequence

Table 3 provides the tabulation of the brachymemma of SEQ ID NO:6 peptide.Shown the influence of brachymemma to the incorporating parametric of these peptides and people FcRn.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

The brachymemma of table 3:SEQ ID NO:6

Table 4 provides the tabulation of SEQ ID NO:1 deutero-peptide and peptide analogs, wherein single amino acids is substituted (L-Ala scanning) with L-Ala.Shown the influence that substitutes the incorporating parametric of these peptides and people FcRn.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

The L-Ala scanning of table 4:SEQ ID NO:1

Table 5 provides the tabulation of SEQ ID NO:1 deutero-peptide and peptide analogs, has wherein carried out with cysteine derivative substituting Gelucystine.Shown the influence that substitutes the incorporating parametric of these peptides and people FcRn.The 1st row contain the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

The cysteine derivative of table 5:SEQ ID NO:1

^*" Pen "=L-Trolovol; " hC "=L-homocysteine;

Table 6 provides the tabulation of SEQ ID NO:1 and peptide No.32 deutero-peptide, and wherein oneself is replaced by the N-methylamino acid with single amino acids.Shown the influence that substitutes the incorporating parametric of these peptides and people FcRn.The 1st row contain the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

The N-methyl scanning of table 6:SEQ ID NO:1 and peptide No.32

^*The Sar=sarkosine; The NMeAla=N-methylalanine; " NMe " prefix designates N-methylamino acid

Table 7 provides the tabulation of the brachymemma of peptide No.32 deutero-peptide derivant.Shown the influence of brachymemma to the incorporating parametric of these peptides and people FcRn.The 1st row contain the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

Table 7: the brachymemma of peptide No.32

^*" Pen "=L-Trolovol

Table 8 provides the tabulation of peptide No.32 deutero-peptide and peptide analogs, wherein under normal circumstances has the place of sequence Gly-Gly-Leu, has produced substituting with different aminoacids and amino acid derivative.The 1st row contain the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

Table 8: peptide No.32 is at the analogue at Gly-Gly-Leu place

^*" Sar "=sarkosine; " Aib "=aminoisobutyric acid

Table 9 provides the tabulation of peptide No.32 deutero-peptide and peptide analogs, wherein under normal circumstances has the place of sequence A rg-Phe-Trolovol, has produced substituting with different aminoacids and amino acid derivative.The 1st row contain the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

Table 9: peptide No.32 is at Arg- PheThe analogue at-Pen place

Table 10 provides the tabulation of peptide No.32 deutero-peptide and peptide analogs, wherein under normal circumstances has the place of sequence Trolovol-Thr-Gly, has produced substituting with different aminoacids and amino acid derivative.The 1st row contain the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

Table 10: peptide No.32 is at Pen- ThrThe analogue at-Gly place

^*" NMeAla "=N-methylalanine

Table 11 provides the tabulation of peptide No.187 deutero-peptide and peptide analogs, wherein under normal circumstances has the place of sequence Phe-Gly-sarkosine, has produced substituting with different aminoacids and amino acid derivative.The 1st row contain the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

Table 11: peptide No.187 is at Pen- GlyThe analogue at-Sar place

^*" Sar "=sarkosine; " Aib "=aminoisobutyric acid

Table 12 provides the tabulation of peptide No.32 deutero-peptide and peptide analogs, wherein under normal circumstances has the place of sequence His-Phe-Gly, has produced substituting with different aminoacids and amino acid derivative.The 1st row contain the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle has shown the chemical structure of Phe analogue side chain.The 4th hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 5th hurdle contains the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes and measures at pH6.

Table 12: peptide No.32 is at His- PheThe analogue at-Gly place

^*" Sar "=sarkosine; " NMeLeu "=N-methylleucine

¹SYN927 is by amido linkage cyclisation between Asp and Lys side chain.

Table 13 provides the tabulation of various peptides and peptide analogs, wherein single amino acids is substituted with tyrosine.Also provide this to substitute influence to the incorporating parametric of these peptides and people FcRn.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle has shown the chemical structure of Tyr analogue side chain.The 4th hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 5th and 6 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

Table 13: tyrosine substitutes

Table 14 provides the tabulation of peptide No.32 deutero-peptide and peptide analogs, wherein under normal circumstances has the place of sequence Gly-Leu, has produced substituting with different aminoacids and amino acid derivative.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

Table 14: peptide No.32 is at the analogue at Gly-Leu place

Table 15 provides the tabulation of peptide No.32 deutero-peptide and peptide analogs, and its place that under normal circumstances has sequence Gly-Leu has glycine and leucine becomes substituting of dipeptide analogue together the time.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle provides the explanation of the identity of X in the sequence.The 4th hurdle has shown the chemical structure of the analogue that is marked as X.The 5th hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 6th hurdle contains the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes and measures at pH6.

Table 15: peptide No.32 is at the simulating peptide analogue at Gly-Leu place

Embodiment 8: peptide synthetic that contains the Histidine analogue

Except the Histidine analogue of following modification, as among the embodiment 7 about the Histidine analogue (table 16) of synthetic modification as described in monomeric peptide disulphide synthetic.Following synthetic peptide No.259, the resin that will contain the complete shielded peptide that is similar to peptide No.99 suspended 15 hours in pure methyl iodide.Clean resin with methylene dichloride, and peptide is cut down from resin, oxidation, and by aforesaid HPLC purifying, to produce the Histidine peptide of monomethylation, i.e. peptide No.259.

Following synthetic peptide No.260, the resin that will contain the complete shielded peptide that is similar to peptide No.99 suspended 72 hours in pure methyl iodide.Clean resin with methylene dichloride, and peptide is cut down from resin, oxidation, and by aforesaid HPLC purifying, to produce two methylated Histidine peptide, i.e. peptide No.260.

Following synthetic peptide No.269, under nitrogen, the resin that will contain the complete shielded peptide that is similar to peptide No.248 suspends in methylene dichloride.With 2,4 of 10 molar equivalents, 6-tri-tert pyridine (Sigma-Aldrich, St.Louis MO) is added into suspension, then adds trifluoromethane-methylmesylate (methyl-trifluoromethane-sulfonate) (Sigma-Aldrich of 5 molar equivalents, St.Louis, MO).Allow be reflected at and carried out in the vibration 4 hours, and, then use the dimethyl formamide rinsing, at last with methylene dichloride rinsing once more with methylene dichloride rinsing for the first time.Peptide is cut down from resin, oxidation, and by aforesaid HPLC purifying, to produce N-methyl-thiazoline (thiazolium) peptide, i.e. peptide No.269.

Following synthetic peptide No.271 is used in and contains 33% ethanol, 10% acetonitrile, 10%N, and 30 equivalent copper sulfate in the solution of the 100mM sodium phosphate buffer pH7.5 of dinethylformamide, 30 equivalent xitix and 10 equivalent sodiumazide are handled peptide No.261.Reaction was carried out 2 hours, and by aforesaid HPLC purifying compounds, comprised the peptide of 1,2,3-triazoles side chain, i.e. peptide No.271 with generation.

Table 16 provides the tabulation of various peptides and peptide analogs, has wherein used single amino acids alternate sets propylhomoserin.The influence that substitutes the incorporating parametric of these peptides and people FcRn also is provided.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle has shown the chemical structure of His analogue side chain.The 4th hurdle contain every kind of peptide IC ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 5th and 6 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

Table 16: Histidine substitutes

Embodiment 9: peptide synthetic that contains the simulating peptide analogue of Gly-Gly

All Gly-Gly amino acid analog things (table 17) mix as the amino shielded amino acid of their Fmoc-, and can obtain by commercial sources, and except as otherwise noted (Chem-Impex, Wood Dale, IL).According to R.M.Freidinger et.al., the described scheme of J.Org.Chem.47:104-109 (1982) is incorporated into peptide No.227 by the N-Fmoc derivative with 3 (R)-3-amino-2-oxygen-1-piperidines-acetate and synthesizes the peptide that contains 3 (R)-3-amino-2-oxo-1-piperidines-acetate.According to R.M.Freidinger et.al., the described scheme of J.Org.Chem.47:104-109 (1982) is mixed peptide No.214 by the N-Fmoc derivative with 3 (R)-3-amino-2-oxygen-1-pyrrolidine acetic acid and is synthesized the peptide that contains 3 (R)-3-amino-2-oxygen-1-pyrrolidine acetic acid.According to N.L.Subasinghe et.al., the described scheme of J.Med.Chem.36:2356-2361 (1993), by with 5,5-dicyclo dipeptide analogue mixes peptide No.197 or peptide No.198 and synthesizes and contain 5, the peptide of 5-dicyclo dipeptide analogue just all uses D-amino acid.According to F.A.Etzkornet.al., the described scheme of J.Am.Chem.Soc.116:10412 (1994), by with 6,5-dicyclo dipeptide analogue mixes peptide No.204 and synthesizes and contain 6, and the peptide of 5-dicyclo dipeptide analogue just all uses D-amino acid.According to R.M.Freidinger et al., the described scheme of J.Org.Chem.47:104-109 (1982), by will (D, L)-the FreidingerShi lactan mixes peptide No.216 and synthesize and contain (D, L)-and the peptide of FreidingerShi lactan, only be to use the L-methionine(Met) to replace the D-methionine(Met).

Table 17 provides the tabulation of SEQ ID NO:1 deutero-peptide and peptide analogs, wherein under normal circumstances has the place of two adjacent glycine (Gly-Gly), has produced substituting with different aminoacids and amino acid derivative.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

Table 17:SEQ ID No:1 is positioned at the analogue at Gly-Gly place

^*" β-Ala "=Beta-alanine; " Apa "=5-aminovaleric acid

Table 18 provides the tabulation of SEQ ID NO:99 deutero-peptide and peptide analogs, and its place that under normal circumstances has sequence Gly-Gly has two glycine becomes substituting of simulating peptide analogue together the time.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle provides the explanation of the identity of the simulating peptide analogue that is marked as X in the sequence.The 4th hurdle has shown the chemical structure of the analogue that is marked as X.The 5th hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 6th hurdle contains the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes and measures at pH6.

Table 18: peptide No.99 is at the simulating peptide analogue at Gly-Gly place

Embodiment 10: the peptide of cyclisation is synthetic by lactam bridges

Solid-phase peptide by general introduction among the embodiment 7 as mentioned synthesizes to come the peptide (table 19) of beta-lactams synthesis cyclisation, only be to use following amino acid substituting: Fmoc-Lys (Aloc)-OH, Fmoc-Orn (Aloc)-OH, Fmoc-Dab (Aloc)-OH and Fmoc-Dap (Aloc)-OH, Fmoc-Glu (OAlly1)-OH and Fmoc-Asp (OAlly1)-OH (Bachem as each halfcystine, Torrance, CA).Finish after this process; promptly on resin, produce after the complete shielded peptide; resin is expanded in methylene dichloride; use nitrogen purge, and with 0.1 molar equivalent tetrakis triphenylphosphine palladium (0) (Sigma-Aldrich, St.Louis; MO) and 30 molar equivalent phenyl silane (Sigma-Aldrich; St.Louis MO) handles, and this reaction was carried out 3 hours.Clean this resin for the first time with methylene dichloride, use DMF then, the last other solution that is used in (v/v) triethylamine of 1% among the DMF and 1% (w/v) diethyldithiocar bamic acid for five times.Carry out extra cleaning step with DMF, then use benzotriazole-1-base-oxygen-three-pyrrolidyl-phosphine phosphofluoric acid ester (PyBOP) (Novabiochem, San Diego CA) and DIEA process resin 16 hours.Embodiment 7 is described as mentioned, and peptide is cut down and purifying from resin.

Table 19 provides the tabulation of different peptide of the present invention, and it has the amino acid replacement that cysteine residues becomes amino acid and amino acid analogue, and this makes the cyclisation by lactam bridges of various peptides.The influence that substitutes the incorporating parametric of these peptides and people FcRn also is provided.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

Table 19: the peptide of lactan cyclisation

¹Indicate between the amino acid whose side chain of underscore and have amido linkage; Dab=1, the 3-DAB; Dap=1, the 2-diaminopropionic acid; The Orn=ornithine

Embodiment 11: linear peptides analogue synthetic

Embodiment 7 described synthesizing linear peptide analogs are the amino acid as table 20 and 21 listed alternative formation disulphide as mentioned.

Table 20 provides the tabulation of SEQ ID NO:1 deutero-linear peptides of the present invention and peptide analogs.The incorporating parametric of these peptides and people FcRn also is provided.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.

The linear analogue of table 20:SEQ ID NO:1

¹" Sar "=sarkosine; " NMeLeu "=N-methylleucine

Table 21 provides the tabulation of peptide No.236 deutero-peptide, wherein under normal circumstances has the place of glycine-sarkosine sequence (Gly-Sar), and oneself has substituted different simulating peptide analogues.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle provides the explanation of the identity of the simulating peptide analogue that is marked as X in the sequence.The 4th hurdle has shown the chemical structure of the simulating peptide analogue that is marked as X.The 5th hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 6th hurdle contains the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes and measures at pH6.

Table 21: the linear analogue of peptide No.236 with Gly-Gly simulating peptide

^*" Sar "=sarkosine; " NMeLeu "=N-methylleucine

Embodiment 12: synthesize peptide dimer by standard reductive alkylation

Standard reductive alkylation by peptide aldehyde and peptide amino (N) or carboxyl (C) terminal amine produces peptide dimer (table 22).

Embodiment 7 synthetic described about monomeric peptide disulphide as mentioned, synthetic peptide N-terminal amine.

Embodiment 7 synthetic described about monomeric peptide disulphide as mentioned, synthetic peptide C-terminal amine, just in synthesis step, use the 1 resin (Novabiochem, San Diego, CA).Therefore, cutting resin produces the terminal ethamine of C-.

When the terminal aldehyde (Fig. 2) of following synthetic peptide N-, DIEA in having DMF, (Sigma-Aldrich, St.Louis MO) reacted 2 hours with the unprotected amino and the 5 equivalent succinyl oxides of-terminal amino acid.Subsequently when having PyBOP and DIEA, with 2,2-dimethyl-1,3-dioxolane methylamine (2,2-dimethyl-1,3-dioxolane methamine) reaction 2 hours produces shielded glycol resin.Then, from resin cutting crude product peptide down, then embodiment 7 synthetic described about monomeric peptide disulphide as mentioned carries out halfcystine oxidation and purifying, generation peptide glycol.Glycol is dissolved in 33% acetate, and (Sigma-Aldrich, St.Louis MO), and carried out reaction 5 minutes then to add 2 equivalent sodium periodates.With 20 equivalents (for glycol) ethylene glycol (Sigma-Aldrich, St.Louis, MO) come the cancellation reaction mixture, after 10 minutes, water is with 3 times of crude product mixture dilutions, and at C18 Sep-Pak post (WatersCrop., Milford MA) go up to use the cumulative gradient of acetonitrile in the water that contains 0.1% TFA to come purifying.With the freeze-drying of peptide aldehyde, and (CA) mass spectrum afterwards (Mariner ES-MS) is to its analysis, as described in embodiment 7 for Applied Biosystems, Foster City to pass through liquid chromatography(LC).

Embodiment 7 synthetic described about monomeric peptide disulphide as mentioned, the terminal aldehyde of synthetic peptide C-only is to use Fmoc-1-amino-2, and (Novabiochem, San Diego CA) replace the Rink amide resins to ammediol-2 '-chlorine trityl resin.Therefore, the gained peptide resin comprises the terminal glycol of masked C-.After cutting down from resin, described about the terminal aldehyde of N-as mentioned, be aldehyde with glycol oxidation.

According to the described methods that are used for synthetic peptide with the lactam bridges cyclisation of embodiment above 10, the synthetic peptide monomer that is used for synthetic peptide such as peptide No.275 with the lactan cyclisation carries out the Asp-Lys cyclisation thus on resin, cut down from resin afterwards.

Following synthetic peptide dimer (Fig. 3), with 1 equivalent peptide aldehyde and 1 equivalent concentration in the DMF that contains 2% acetate be 40mg/ml contain the amine reactive polypeptide.After 60 minutes, (Sigma-Aldrich, St.Louis MO), and make reaction vibration 1 hour to add 2 equivalent sodium cyanoborohydrides.Water is with 10 times of reaction mixture dilutions, and by the HPLC purifying, and (CA) mass spectrum afterwards (Mariner ES-MS) is analyzed, as described in embodiment 7 for Applied Biosystems, Foster City to pass through liquid chromatography(LC).

Table 22 provides the tabulation by standard reductive alkylation synthetic dimerization peptide of the present invention.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.The 6th hurdle contains the IC of every kind of peptide ₅₀, its competitive IgG by general introduction among the embodiment 5 measures in conjunction with facs analysis.

Table 22: by standard reductive alkylation synthetic dimer and tripolymer

X=3 (r)-3-amino-1-carboxylic formic acid-Valerolactim

Embodiment 13: by mercaptan joint and the synthetic peptide dimer of acetobrom peptide

Also can react to synthesize peptide dimer (table 23) by acetobrom peptide and mercaptan joint.With the free alpha-amino group of shielded peptide resin and 4 equivalent bromoacetyl bromides among the DMF (Sigma-Aldrich, St.Louis, MO) and 8 equivalent DIEA (MO) reaction comes the synthetic bromide acetylated peptide for Sigma-Aldrich, St.Louis.After 1 hour, resin is cleaned with DMF, then clean, and embodiment 7 is described as mentioned, cuts down from resin with DCM.In using the situation of two mercaptan joint dimerizations, before carrying out the acetobrom step, carry out the cyclisation step on the resin with the peptide of lactan cyclisation.Using two mercaptan joint dimerizations to contain in the situation of peptide of disulphide, embodiment 7 is described as mentioned, carries out the iodine oxidation step after cutting.

Following synthetic two mercaptan joints, (Novabiochem, SanDiego is during CA) with DIEA, with NH for 2 equivalent PyBOP in having DMF ₂-Gly-2-chlorine trityl resin (Novabiochem, SanDiego, CA) with 2 equivalent N, N-two (N '-the Fmoc-3-aminopropyl) (IL) reaction is 18 hours for Chem-lmpex, Wood Dale for glycine hemisulfic acid potassium.The processing of carrying out 2 times 10 minutes with 20% piperidines among the DMF is to remove the Fmoc blocking group.For some linker compounds, also mix Beta-alanine as interval unit.Use PyBOP and DIEA as mentioned, Fmoc-β-Ala-OH (Novabiochem) is coupled to resin.Behind 20% piperidines removal Fmoc blocking group among the DMF, or mix another Beta-alanine unit at interval, or by N-terminal amine resin and the 2 equivalent N-succinimido-S-acetyl thio propionic ester (SATP that will dissociate; Pierce, Rockford, IL) two mercaptan joints were mixed in reaction in 18 hours with 4 equivalent DIEA.

Subsequently; by with the 0.05mmol peptide resin with contain 1ml DMF and 0.4ml buffer A (buffer A: 1M azanol hydrochloric acid (Sigma-Aldrich; St.Louis; MO), 40mM sodium phosphate pH7.5,50mMEDTA (Sigma-Aldrich; St.Louis, MO)) de-gassed solution reaction realized the removal of S-ethanoyl blocking group in 18 hours.Resin is cleaned with DMF, then cleans with DCM, and with 50% TFA solution among the DCM that contains 2% tri isopropyl silane with resin cutting 15 minutes.Embodiment 7 is described as mentioned, processing and purifying crude product joint.

Following use two mercaptan joints produce peptide dimer (Fig. 4), and the two mercaptan joints and the 2 equivalent acetobrom N-terminal peptide of 1 equivalent purifying are reacted in the DMF that contains 10% water and 50%100mM sodium phosphate pH7.5.After 18 hours, embodiment 7 is described as mentioned, by reversed-phase HPLC column purification crude product reaction mixture.

Following synthetic peptide No.122 (Fig. 5) is with acetobrom peptide and the reactive polypeptide of using the SATP derivatize.In brief, the crude product peptide resin and the SATP of 2 equivalents in DMP that will have free N-terminal amine reacted 2 hours.Remove S-ethanoyl blocking group as mentioned above, then cut down subsequent purificn as mentioned above from resin.

Table 23 provides the tabulation by mercaptan joint synthetic dimer peptide of the present invention.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.The 4th and 5 hurdles contain the K of every kind of peptide _D, its Biacore by general introduction among the embodiment 6 analyzes respectively and measures at pH6 and pH7.4.The 6th hurdle contains the IC of every kind of peptide ₅₀, its competitive IgG by general introduction among the embodiment 5 measures in conjunction with facs analysis.

Table 23: use mercaptan joint synthetic dimer

¹The Pen=Trolovol; The Suc=succsinic acid; The Sar=sarkosine; The p=D-proline(Pro); X=(3R)-3-amino-1-carboxymethyl Valerolactim; The NMeLeu=N-methylleucine

Embodiment 14: synthesize the peptide tripolymer by standard reductive alkylation: peptide No.247

By standard reductive alkylation peptide aldehyde and peptide amino N-terminal amine, produce peptide tripolymer (table 22).

As among the embodiment 7 about as described in monomeric peptide disulphide synthetic, synthetic peptide N-terminal amine, just N-terminal with difunctional amine joint such as diamines propyl group glycine (bis-aminipropylglycine) (BAPG; Use with the Bis-Fmoc-BAPG form, it is available from Sigma-Aldrich, and StI.Louis MO) adds cap, then the coupling sarkosine.As described in embodiment 12, the terminal aldehyde (Fig. 2) of synthetic peptide N-.Following synthetic peptide tripolymer (Fig. 3), with 2 equivalent peptide aldehyde and 1 equivalent the concentration in the DMF that contains 2% acetate be 40mg/ml contain the amine reactive polypeptide.After 60 minutes, (Sigma-Aldrich, St.Louis MO), make reaction vibration 1 hour then to add 4 equivalent sodium cyanoborohydrides.Water is with 10 times of reaction mixture dilutions, and by the HPLC purifying, and (CA) mass spectrum afterwards (Mariner ES-MS) is analyzed, as described in embodiment 7 for Applied Biosystems, Foster City to pass through liquid chromatography(LC).

Embodiment 15: use the synthetic peptide dimer of diacid and amine joint

Or the terminal and difunctional sour joint reaction of N-of two peptide monomers by will be on resin, or by on the resin that contains difunctional amine joint, carrying out the synthetic of peptide, the C-end of two peptide monomers that thus will be on resin connects together, thereby produces the peptide dimer (table 24) that acid amides connects.

Embodiment 7 synthetic described about monomeric peptide disulphide as mentioned, the terminal peptide dimer that connects of synthetic N-, just: before peptide under the resin cutting, be connected with difunctional sour joint the N-of two peptide monomers is terminal.For example, following synthetic peptide No.283 when having 1 equivalent PyBOP and 2 equivalent DIEA, will contain the peptide resin and 0.5 equivalent succsinic acid (Sigma-Aldrich, St.Louis, MO) reaction that are similar to the peptide sequence with the peptide No.235 that does not protect the N-end.This cause on the resin contiguous peptide by their N-ends through amido linkage and covalent attachment.

Embodiment 7 is described as mentioned, and the gained peptide dimer is cut down and purifying from resin, and just the HPLC purifying is not before with the oxidation of peptide disulphide.The phthalin of purifying is dissolved to about 0.1mg/mL in containing the 10mM sodium phosphate pH7.5 solution of 20% DMSO, and in mixed at room temperature 3 days.This oxidation step forms disulfide linkage in a dimeric peptide monomer, with the difference mutually of formation between dimeric two monomers.Water is diluted to the peptide concentration of 0.05mg/mL with reaction mixture, and (Waters Corp., Milford MA) go up the cumulative gradient of using in the water that contains 0.1% TFA of acetonitrile and be purified at C18 Sep-Pak post.With the peptide dimer freeze-drying, and by liquid chromatography(LC) (CA) mass spectrum afterwards (Mariner ES-MS) is analyzed for Applied Biosystems, Foster City, as (see figure 6) as described in the embodiment 7.In the situation of peptide No.283, to peptide digestion 30 minutes,, thereby confirm the disulfide linkage pattern then with lcms analysis gained peptide by trypsinase.Known trypsinase cuts at arginine and lysine residue back, and at arginine-phenylalanine key place cutting peptide No.283.The LCMS primary product of peptide No.283 is NH ₂-[Phe-Phe-Pen-Thr-Gly-His-Phe-Gly-Sar-NmeLeu-Tyr-Pro-Cys]-CONH ₂(disulphide) (LCMS:M+H=1355.6Da), this disulfide linkage that shows peptide No.283 forms between each 13 amino acid whose peptide monomer.

Synthetic peptide No.201 as peptide No.283, just peptide sequence and peptide No.32 are similar, and the diacid joint of use is ethylene glycol-two (succsinic acid-N-hydroxy-succinamide ester) (Sigma-Aldrich, St.Louis, MO), and PyBOP is not used for linked reaction.

Synthetic peptide No.279 as peptide No.283, the diacid joint that only is to use is Bis-dPEG6-N-hydroxysuccinimide eater (Quanta Biodesigns Ltd.), and PyBOP is not used for linked reaction.

Synthetic peptide No.281 as peptide No.283, just (Sigma-Aldrich, St.Louis MO) handle peptide-resin, and this causes all peptides on resin all to contain the N-end that succinate adds cap with excessive greatly succinyl oxide.When having 1 equivalent PyBOP and 2 equivalent DIEA, with 0.5 equivalent N, N '-dimethyl-ethylenediamine (Sigma-Aldrich, St.Louis, MO) process resin.As to peptide No.283, carry out subsequently cutting, purifying and oxidation step.

Synthetic peptide No.282 as peptide No.283, the diacid joint that only is to use be N-methyl-iminodiethanoic acid (Sigma-Aldrich, St.Louis, MO).

Synthetic peptide No.284 as peptide No.283, the diacid joint that only is to use is 3, the 3-dimethylated pentanedioic acid (Sigma-Aldrich, St.Louis, MO).

Synthetic peptide No.285 as peptide No.283, the diacid joint that only is to use be Boc-Asp (OH)-OH (Novabiochem, San Diego, CA).

Synthetic peptide No.286 as peptide No.283, the diacid joint that only is to use be Boc-Glu (OH)-OH (Novabiochem, San Diego, CA).

Embodiment 7 just was coupled to resin with difunctional amine joint about the synthetic described terminal peptide dimer that connects of C-that synthesizes of monomeric peptide disulphide before peptide is synthetic as mentioned.This causes peptide dimer by the C-end covalent attachment of amido linkage with them.For example, following synthetic peptide No.280, at first (Novabiochem, San Diego CA) are coupled to resin, and then coupling amino acid is to provide the similar sequence with peptide No.235 with Fmoc-Lys (Fmoc)-OH.When they were synthetic on resin, this caused the covalent attachment of two peptide chains.The gained peptide dimer is cut down from resin, purifying and oxidation, described about the terminal dimer that connects of N-as mentioned.(see figure 7)

Synthetic peptide No.287 as peptide No.280, just glycine residue (Gly) inserts between peptide No.235 sequence and branch's Methionin joint.

Synthetic peptide No.288 as peptide No.280, just two glycine residues (Gly-Gly) insert between peptide No.235 sequence and branch's Methionin joint.

Table 24 provides the tabulation of using the synthetic dimerization peptide of the present invention of amido linkage.The 1st hurdle contains the peptide identifier.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.

Table 24: use acid amides joint synthetic dimer

¹The Pen=Trolovol; The Sar=sarkosine; The NMeLeu=N-methylleucine

Embodiment 16: by the synthetic peptide of standard reductive alkylation-Fc fusions

As described in embodiment 12, synthetic peptide N-terminal aldehyde, i.e. peptide No.252, peptide No.229 and peptide No.232 (table 25).All three kinds of peptides-Fc fusions use identical scheme to produce (Fig. 8): with CysFc (the Fc structural domain that has the N-terminal cysteine) and 4.5 normal peptide aldehyde in 80mM sodium acetate pH5.5 incubation on ice 1 hour.Add sodium cyanoborohydride to final concentration 20mM, and will react on 4 ℃ of incubations 16 hours.Come analyze reaction mixture to guarantee it mainly is that single peptide is added into Fc protein by SDS-PAGA.With PBS protein mixture is cleaned 2 times, and measure extracorporeal blocking activity (table 25).In the situation of peptide No.252-Fc, protein is also assessed (Figure 15) in TG32B mouse IgG katabolism model.The production of CysFc can be carried out as described in U.S. Patent application No.US 2005/0027109, with production imcome this paper of disclosed CycFc wherein as a reference.

Table 25 provides the tabulation of using CysFc and aldehyde-peptide synthetic peptide of the present invention-Fc fusion rotein.The 1st hurdle contains the identifier of peptide-Fc fusions.The 2nd hurdle contains the aminoacid sequence of peptide.The 3rd hurdle contains the IC of every kind of peptide ₅₀, its IgG competitive ELISA by general introduction among the embodiment 4 is measured.

Table 25: peptide-Fc fusions

¹The Pen=Trolovol; The Sar=sarkosine; The NMeLeu=N-methylleucine

Embodiment 17: transgenic mice

Transgenic mice available from doctor Roopenian (Jackson Laboratory, Bar Harbor, ME).Endogenous murine FcRn and β ₂The m gene inserts external polynucleotide sequence and deactivation by homologous recombination, and personnel selection FcRn and people β ₂The transgenosis replacement of m gene (muFcRn (/-), mu β ₂M (/-) ,+huFcRn ,+hu β ₂M).These mouse are called with strain name TG32B.

Embodiment 18:5mg/kg and 10mg/kg peptide No.270 are to the catabolic influence of human IgG in the TB32B mouse

At t=0 hour (T ₀) time, to injection 500mg/kg human IgG in the TG32B mouse vein of growing up (MPBiomedicals, Irvine, CA).At 24 hours, 48 hours, 72 hours, 96 hours and 120 hours, to injection 5mg/kg or 10mg/kg peptide No.270 in the mouse vein.At each time point, use the media PBS pH5 that contains the 15mM sodium acetate to contrast injection.All time points and 168 hours, blood sample collection before injection.Preparation serum also is stored in-20 ℃, until carrying out ELISA (Fig. 9).

At each time point, IgG Fc structural domain specific ELISA is used for detecting the human IgG level of serum.In brief, with 6ml 0.05M sodium bicarbonate pH9.6 (Sigma-Aldrich, St.Louis, MO) the anti-human IgG storing solution of dilution 30 μ l, 10 μ g/ml goats (Pierce, Rockford, IL).With this solution bag in 50 μ l/ holes by 96 orifice plates, and in 37 ℃ of incubations 1 hour.To wrap, and clean once with PBST (phosphate buffered saline (PBS) that contains 0.05% tween 20) by solution removal.Add 2% bovine serum albumin(BSA) (BSA) storing solution of 200 μ l/ holes in PBS then, and with flat board in 37 ℃ of incubations 1 hour.With PBST the hole is cleaned 3 times, and produced typical curve by 2.5 times of dilutions that start from 50ng/ml hIgG1 in triplicate.Then 100 μ l standard substance or sample solution are added into each hole, and with flat board in 37 ℃ of incubations 1 hour.Carry out 3 PBST again and clean, then add 100 μ l 1:10 in the PBS that contains 2% BSA, and the anti-human IgG of goat [the Fc]-HRP conjugate of 000 dilution (Pierce, Rockford, IL).Make and dull and stereotyped then clean with PBST in 37 ℃ of incubations 1 hour, and in each hole, add 100 μ l TMB single component substrates (BioFX, Owings Mills, MO).After 5 minutes, come color development stopping by in each hole, adding 100 μ l 0.25M sulfuric acid.At the 450nm place, measure the UV absorbancy in each hole, and derive the graphic representation of the serum IgG concentration of each experiment the time with working curve.

Embodiment 19: peptide No.231, peptide No.274 and peptide No.252-Fc are to the catabolic influence of human IgG in the TG32B mouse

At t=0 hour (T ₀) time, to injection 500mg/kg human IgG in the TG32B mouse vein of growing up (MPBiomedicals, Irvine, CA).At 24 hours, 48 hours and 72 hours, to injection 1mg/kg peptide No.231,1mg/kg peptide No.274 or 20mg/kg peptide No.252-Fc in the mouse vein.At each time point, use 15mM sodium acetate pH5 to contrast injection, and with its media as all injections.All time points and 30 hours, 96 hours and 144 hours, blood sample collection before injection.Preparation serum also is stored in-20 ℃, until carrying out ELISA.

Embodiment 18 is described as mentioned, the human IgG concentration (Figure 15) in each time point determining serum.Embodiment 20: peptide No.270 is to the human IgG katabolism in the macaque and endogenous IgG, IgM and albuminised influence

At t=0 hour (T ₀) time, to 3 mean body weights be 4.8kg adult macaque intravenous injection vein dosage (IV dose) for the biotinylation human IgG of 5mg/kg (MP Biomedicals, Irvine, CA).At 24 hours, 48 hours, 72 hours and 96 hours, to this animal with the speed intravenous injection 10mg/kg peptide No.270 of 1ml/min or isopyknic media (the 30mM sodium acetate, pH5).At 120 hours, handle animal CO6215 with the 5th dose of 10mg/kg peptide No.270.Before all injections and 120 hours, 168 hours, 192 hours and 244 hours with at the 30th day, blood sample collection before injecting.Preparation serum also is stored in-20 ℃, until carrying out ELISA (Figure 10-12).

Use strepto-affinity element-Fc specific ELISA to come detection of biological element-hIgG tracer.(IL cat#15121) cleans 3 times for Pierce, Rockford with the flat board of the plain bag of strepto-affinity quilt with PBST (phosphate buffered saline (PBS)+0.05% tween 20).Come dilute serum sample and standard substance with PBSB (PBS+2% BSA).Typical curve is established at 1.56ng/ml on the scope of 200ng/ml.The sample (100 μ l) or the standard substance of dilution are added into each hole, and in room temperature incubation 2 hours.Then, with PBST (300 μ l/ hole) hole is cleaned 3 times.With PBSB with the anti-people Fc-HRP of goat (Pierce, Rockford, IL, cat#31416) dilution 1:25,000, add 100 μ l/ holes, and with flat board in room temperature incubation 30 minutes.With PBST (300 μ l/ hole) flat board is cleaned 3 times, and (MD) colour developing is about 5 minutes for BioFx, Owing Mills with 100 μ l/ hole BioFx Supersensitive tmb substrates in room temperature.Stop color reaction by adding 100 μ l/ hole 0.25M sulfuric acid, and measure the absorbancy in each hole at 450nm wavelength place.

Use following ELISA scheme to detect endogenous macaque IgG.At first, (bag is cushioned in liquid=1 carbonate-supercarbonate capsule (Sigma-Aldrich, St.Louis MOcat#C-3041) are dissolved in 100ml water) and is diluted to 2 μ g/ml the anti-monkey IgG of rabbit to be cushioned liquid at bag.Then, with 96 orifice plates (Costar/Corning) with the anti-monkey IgG of 100 μ l/ holes, 2 μ g/ml rabbits (Sigma-Aldrich, St.Louis, MO) bag quilt, and in 37 ℃ of incubations 1 hour.Clean dull and stereotyped 4 times with PBST (PBS that contains 0.05% tween 20), and with 200 μ l/ hole PBSB (the PBS solution of 1%BSA; Dilute from the PBS of 10% BSA storing solution; KPL) in 37 ℃ of sealings 1 hour.With PBST flat board is cleaned 4 times again.With PBSB dilute serum sample and standard substance.(Antibodies Incorporated, Davis is CA) on the scope to 1.9ng/ml monkey IgG at 2000ng/ml with the typical curve establishment.Then with the every kind of sample in 100 μ l/ holes in 37 ℃ of incubations 1 hour.Clean dull and stereotyped 3 times with PBST.Add 100 μ l/ hole 1:30 in PBSB, the anti-monkey IgG-HRP of rabbit of 000 dilution (Sigma-Aldrich, St.Louis, MO), and in 37 ℃ of incubations 1 hour.Clean dull and stereotyped 3 times with PBST, and (MD) colour developing is about 5 minutes for KPL, Gaithersburg with 100 μ l/ hole SureBlue tmb substrates in room temperature.(KPL, Gaithersburg MD) stop color reaction, and measure the absorbancy in each hole at 450nm wavelength place to stop solution with 100 μ l/ hole TMP.

Use following ELISA scheme to detect endogenous serum of macaque white protein.At first, the anti-monkey serum white protein of rabbit is cushioned in the liquid (bag is cushioned liquid=1 carbonate-supercarbonate capsule (Sigma-Aldrich, St.Louis, MO cat#C-3041) and is dissolved in 100ml water) at bag is diluted to 5 μ g/ml.Then, with 96 orifice plates (Costar/Corning) with the anti-monkey serum white protein of 100 μ l/ holes, 5 μ g/ml rabbits (Nordic Immunology, TheNetherlands, cat#RAMon/Alb) bag quilt, and in 37 ℃ of incubations 1 hour.Clean dull and stereotyped 4 times with PBST (PBS that contains 0.05% tween 20), and sealed 1 hour in 37 ℃ with 5% isinglass (Sigma-Aldrich, St.Louis, the MO cat#G-7765) storing solution of 300 μ l/ holes in PBS.With PBST flat board is cleaned 4 times again.With PBSB dilute serum sample and standard substance.(Nordic Immunology, The Netherlands is cat#RAMon/Alb) on the scope to 0.39ng/ml monkey serum white protein at 200ng/ml with the typical curve establishment.Then with the every kind of sample in 100 μ l/ holes in 37 ℃ of incubations 1 hour.Clean dull and stereotyped 6 times with PBST.Add 100 μ l/ hole 1:30 in PBSB, the anti-people's white protein of the goat-HRP conjugate of 000 dilution (Academy Bio-Medical, Inc., Houston, TX, cat#AL10H-Gla), and in 37 ℃ of incubations 1 hour.Clean dull and stereotyped 6 times with PBST, and (MD) colour developing is about 5 minutes for KPL, Gaithersburg with 100 μ l/ hole SureBlue tmb substrates in room temperature.(KPL, Gaithersburg MD) stop color reaction, and measure the absorbancy (rabbit 13) in each hole at 450nm wavelength place to stop solution with 100 μ l/ hole TMP.

Use following ELISA scheme to detect endogenous macaque IgM.At first, the anti-monkey IgM of goat antibody is cushioned in the liquid (bag is cushioned liquid=1 carbonate-supercarbonate capsule (Sigma-Aldrich, St.Louis, MO cat#C-3041) and is dissolved in 100ml water) at bag is diluted to 5 μ g/ml.Then, with 96 hole flat boards (Costar/Corning) with the anti-monkey IgM of 100 μ l/ holes, 5 μ g/ml goats (KPL, Gaithersburg, MD, cat#071-11-031) bag quilt, and in 37 ℃ of incubations 1 hour.Clean dull and stereotyped 4 times with PBST (PBS that contains 0.05% tween 20), and with 200 μ l/ hole PBSB (the PBS solution of 1%BSA; Dilute from the PBS of 10% BSA storing solution; KPL) in 37 ℃ of sealings 1 hour.With PBST flat board is cleaned 4 times again.With PBSB dilute serum sample and standard substance.(TX is cat#2001301) on the scope for Alpha Diagnostic International, San Antonio to 15.6ng/ml monkey IgM at 2000ng/ml with the typical curve establishment.Then with the every kind of sample in 100 μ l/ holes in 37 ℃ of incubations 1 hour.With PBST flat board is cleaned 3 times.Add 100 μ l/ hole 1:10 in PBSB, the anti-monkey IgM-HRP of the goat conjugate of 000 dilution (RDI, Concord, MA, cat#617103007), and in 37 ℃ of incubations 1 hour.Clean dull and stereotyped 4 times with PBST, and with 100 μ l/ hole SureBlue tmb substrates (KPL, Gaithersburg, MD) in color development at room temperature about 5 minutes.(KPL, Gaithersburg MD) stop color reaction, and measure the absorbancy (Figure 14) in each hole at 450nm wavelength place to stop solution with 100 μ l/ hole TMP.

Embodiment 21: peptide No.270 is to the catabolic influence of human IgG in the TG32B mouse when using the various dose dosage regimen

At t=0 hour (T ₀) time, to injection 500mg/kg human IgG in the TG32B mouse vein of growing up (MPBiomedicals, Irvine, CA).In the time of t=24 hour, to injection 5mg/kg peptide No.270 in first group of four mouse vein; T=24 and 72 hours, to injection 5mg/kg peptide No.270 in second group of four mouse vein; T=24,48,72,96 hours, to injection 2.5mg/kg peptide No.270 in the 3rd group of four mouse veins.At each time point, use the media PBSpH5 that contains the 15mM sodium acetate to contrast injection to other one group of mouse.At all time points and at 168 hours, blood sample collection before injection.Preparation serum also is stored in-20 ℃, until carry out ELISA (Figure 16) as described in embodiment 18.

Embodiment 22: other TG32B mouse experiment

In the TG32B mouse, carry out other experiment with peptide No.270.Use and embodiment 18 described identical experimental designs, find to use subcutaneous (SC) and intraperitoneal (IP) when using the path, peptide No.270 effectively quickens the catabolic speed of IgG.After subcutaneous (SC) and intraperitoneal (IP) injection peptide No.270, find from 5 doses of beginning in 24 hours once a day 5mg/kg peptide No.270 the transformation period of IgG was reduced to 56 hours.These transformation period are significantly shorter than typical control group, and it shows 80 to 100 hours IgG transformation period.In addition, compare with control group, after using peptide No.270 168 hours, the concentration of hIgG reduced by 56% (SC) and 66% (IP).

Also use embodiment 18 described experimental programs in the TG32B mouse, to test peptide No.230.Behind the intravenous injection human IgG 24 hours, used intravenous injection 5mg/kg peptide No.230 once a day totally in five days.Compare with 92 hours transformation period of control group, the half life of hIgG, be reduced to 39 hours.In addition, compare with control group, the hIgG concentration after 168 hours has reduced by 76%.

Also be designed in the single agent peptide dosage of assessment and the 3 doses of experiments of the effect compared of peptide dosage once a day and tested peptide No.230 at two.Use embodiment 18 described experimental programs, behind the intravenous injection human IgG 24 hours, first animal groups is handled with single intravenous dosages 5mg/kg peptide No.230, and second animal groups accepted three successive intravenous dosages 5mg/kg peptide No.230 once a day.After 120 hours, single dosage peptide No.230 has reduced by 41% with the hIgG concentration in the mouse.Accepting 3 once a day in the mouse group of dosage peptide No.230, the hIgG concentration after 120 hours has reduced by 61%.

Embodiment 23: peptide No.283 is to the catabolic influence of human IgG in the TG32B mouse

At t=0 hour (T ₀) time, to injection 500mg/kg human IgG in the TG32B mouse vein of growing up (MPBiomedicals, Irvine, CA).At 24 hours, 48 hours, 72 hours and 96 hours, to injecting 0.5,1,2.5,5 or 10mg/kg peptide No.283 in the mouse vein.At each time point, use 15mM sodium acetate pH5 to contrast injection, and with its media as all injections.At all time points and at 120 hours, 168 hours with at the 30th day, blood sample collection before injection.Preparation serum also is stored in-20 ℃, until carrying out ELISA.

Embodiment 18 is described as mentioned, measures the human IgG concentration (Figure 17) in each time point serum.Embodiment 24:PEGization peptide No.289's is synthetic

Peptide No.285 is dissolved in 10mM phosphoric acid salt pH7.4 damping fluid, and with 1 equivalent PEG _30kDa-succinimido ester (NOF Corp, Japan, Sunbright MEGC-30TS) was handled 18 hours.As described in embodiment 7, (Jupiter Phenomenex) goes up purifying, freeze-drying, and with cation-exchange chromatography (Fractoprep SO at the C4 post with the crude product reaction mixture ₃ ^-, Cat No.1.17972, EMD Chemicals Inc, Gibbstown, NJ) purifying once more, peptide is bonded to the resin among the 10mM sodium acetate pH5 thus, and pH5 cleans resin with the 10mM sodium acetate, and with the 100mM sodium-chlor wash-out peptide among the 10mM sodium acetate pH5.Peptide solution is dialysed at 1% acetate, and freeze-drying.Peptide to purifying is analyzed, and SDS-PAGE shows peptide dyeing band at about 50KDa place, and HPLC shows the free peptide (Figure 18) that does not have remnants.

Table 26: the PEGization analogue of peptide No.285

Embodiment 25: peptide No.289 is to the catabolic influence of human IgG in the TG32B mouse

At t=0 hour (T ₀) time, to injection 500mg/kg human IgG in the TG32B mouse vein of growing up (MPBiomedicals, Irvine, CA).At 24 hours, to injection 25mg/kg peptide No.289 in the mouse vein.At 24,48,72,96,120 and 168 hours, blood sample collection.Preparation serum also is stored in-20 ℃, until carrying out ELISA.Embodiment 18 is described as mentioned, measures the human IgG concentration (Figure 19) in each time point serum.

Embodiment 26: peptide No.283 is to the influence of hIgG katabolism in the macaque and endogenous IgG, IgM and white protein concentration

18 macaques are divided into 6 groups, i.e. every group of 3 animals, and in the time of t=-3 days, handle all animals with 5mg/kg biotinylation human IgG (MPBiomedical).From t=0,, animal was handled for 4 weeks: 1) intravenously 1mg/kg 3x/ week with peptide No.28 according to following dosage regimen; 2) subcutaneous 1mg/kg1x/ week; 3) subcutaneous 1mg/kg 3x/ week; 4) intravenously 5mg/kg 3x/ week; 5) subcutaneous 5mg/kg 1x/ week; 6) subcutaneous 5mg/kg 3x/ week.Notice that the 4th group last potion peptide is at the 16th day.Gathered serum sample at-3 days ,-15 minutes, 1 day, 2 days, 3 days, 4 days, 5 days, 7 days, 9 days, 11 days, 14 days, 16 days, 18 days, 21 days, 23 days, 25 days, 28 days, 30 days, 32 days, 35 days, 42 days, 49 days, 77 days.As described in embodiment 20, measure biotinylation human IgG, endogenous IgG and albuminised concentration, and be shown in Figure 20-24.

According to the instruction of the reference of quoting in this specification sheets, can understand this specification sheets the most thoroughly.Embodiment in the specification sheets provides the example of embodiment of the present invention, the not scope that should be construed as limiting the invention.The technician can be easy to recognize that many other embodiments are contained in the present invention.All publications quoted in the disclosure and the complete income of patent are as a reference.Every income material and this specification sheets contradiction or inconsistent as a reference just replaces any this material with this specification sheets.Here quoting of any reference is not to recognize that these reference are prior aries of the present invention.

Except as otherwise noted, this specification sheets comprises that all numerical value that are used for being expressed as dosis refracta, reaction conditions or the like in claims should be understood to: it is modified with term " about " at all situations.Thereby unless opposite explanation is arranged in addition, numerical parameter is approximate the finger, and the desired characteristic that can attempt to obtain along with the present invention and difference.At least, and the application of the doctrine that is not equal to as the scope of attempting to limit with claim, each numerical parameter should be explained according to significant figure and conventional rounding method.

Except as otherwise noted, the term before a series of compositions " at least " is interpreted as the every kind of composition that refers in the series.Those skilled in the art only are to use routine test just to be familiar with or to understand fully, the many modes that are equal to specific embodiments of the present invention described herein.Intention is encompassed in this equivalent way in the claims.

Sequence table

＜110〉Syntonix Pharmaceuticals Inc. (SYNTONIX PHARMACEUTICALS, INC.)

＜120〉blocking-up IgG is to the bonded peptide of FcRn

<130>08945.0017-00304

<140>

<141>

<150>60/774,853

<151>2006-02-17

<150>60/805,634

<151>2006-06-23

<160>322

<170>PatentIn?version?3.3

<210>1

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>1

<210>2

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>2

<210>3

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>3

<210>4

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>4

<210>5

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>5

<210>6

<211>25

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>6

<210>7

<211>24

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>7

<210>8

<211>25

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>8

<210>9

<211>25

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>9

<210>10

<211>25

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>10

<210>11

<211>25

<212>DNA

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic primer

<400>11

<210>12

<211>26

<212>DNA

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic primer

<400>12

<210>13

<211>26

<212>DNA

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic primer

<400>13

<210>14

<211>7

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(6)..(6)

＜223〉variable amino acid

<400>14

<210>15

<211>1098

<212>DNA

＜213〉human (Homo sapiens)

<400>15

<210>16

<211>360

<212>DNA

＜213〉human (Homo sapiens)

<400>16

<210>17

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>17

<210>18

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>18

<210>19

<211>11

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>19

<210>20

<211>9

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>20

<210>21

<211>16

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>21

<210>22

<211>15

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>22

<210>23

<211>16

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>23

<210>24

<211>15

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>24

<210>25

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>25

<210>26

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>26

<210>27

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>27

<210>28

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>28

<210>29

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>29

<210>30

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>30

<210>31

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>31

<210>32

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>32

<210>33

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>33

<210>34

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>34

<210>35

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>35

<210>36

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>36

<210>37

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(14)..(14)

<223>homocysteine

<400>37

<210>38

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

＜223〉homocysteine

<220>

<221>MOD_RES

<222>(14)..(14)

＜223〉homocysteine

<400>38

<210>39

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>D-Cys

<400>39

<210>40

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(14)..(14)

<223>D-Cys

<400>40

<210>41

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>D-Cys

<220>

<221>MOD_RES

<222>(14)..(14)

<223>D-Cys

<400>41

<210>42

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>42

<210>43

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(14)..(14)

<223>Pen

<400>43

<210>44

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(14)..(14)

<223>Pen

<400>44

<210>45

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(14)..(14)

＜223〉homocysteine

<400>45

<210>46

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

＜223〉homocysteine

<220>

<221>MOD_RES

<222>(14)..(14)

<223>Pen

<400>46

<210>47

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<220>

<221>MOD_RES

<222>(14)..(14)

<223>Pen

<400>47

<210>48

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(5)..(5)

<223>NMeAla

<400>48

<210>49

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>49

<210>50

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

<223>Sar

<400>50

<210>51

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

<223>NMeHis

<400>51

<210>52

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>NMePhe

<400>52

<210>53

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<400>53

<210>54

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>Sar

<400>54

<210>55

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(11)..(11)

<223>NMeLeu

<400>55

<210>56

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(12)..(12)

<223>NMeTyr

<400>56

<210>57

<211>16

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>57

<210>58

<211>15

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<400>58

<210>59

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>Sar

<220>

<221>MOD_RES

<222>(11)..(11)

<223>NMeLeu

<400>59

<210>60

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>60

<210>61

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>61

<210>62

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(2)..(2)

<223>Pen

<400>62

<210>63

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(2)..(2)

<223>Pen

<400>63

<210>64

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<400>64

<210>65

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>65

<210>66

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Pro

<400>66

<210>67

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(2)..(2)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>D-Pro

<400>67

<210>68

<211>16

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Pro

<400>68

<210>69

<211>15

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(2)..(2)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>D-Pro

<400>69

<210>70

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>70

<210>71

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>71

<210>72

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Arg

<400>72

<210>73

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-His

<400>73

<210>74

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Ile

<400>74

<210>75

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Phe

<400>75

<210>76

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Tyr

<400>76

<210>77

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>Aib

<400>77

<210>78

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Asp

<400>78

<210>79

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Pro

<400>79

<210>80

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Arg

<400>80

<210>81

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-His

<400>81

<210>82

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Ile

<400>82

<210>83

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Phe

<400>83

<210>84

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Tyr

<400>84

<210>85

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Aib

<400>85

<210>86

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>M01D_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Ala

<400>86

<210>87

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Ala

<400>87

<210>88

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(10)

<223>D-Ala

<400>88

<210>89

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Ala

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>89

<210>90

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Phe

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>90

<210>91

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Phe

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Ala

<400>91

<210>92

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(10)

<223>D-Pro

<400>92

<210>93

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Phe

<220>

<221>MOD_RES

<222>(11)..(11)

<223>NMeLeu

<400>93

<210>94

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Ala

<220>

<221>MOD_RES

<222>(11)..(11)

<223>NMeLeu

<400>94

<210>95

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Phe

<400>95

<210>96

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Pro

<400>96

<210>97

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Phe

<400>97

<210>98

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Ala

<220>

<221>MOD_RES

<222>(10)..(10)

<223>Sar

<400>98

<210>99

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>99

<210>100

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>D-Phe

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>100

<210>101

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>101

<210>102

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>102

<210>103

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>103

<210>104

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>104

<210>105

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>105

<210>106

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(5)..(5)

<223>NMeAla

<400>106

<210>107

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>Sar

<220>

<221>MOD_RES

<222>(11)..(11)

<223>NMeLeu

<400>107

<210>108

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Ala

<220>

<221>MOD_RES

<222>(10)..(10)

<223>Sar

<220>

<221>MOD_RES

<222>(11)..(11)

<223>NMeLeu

<400>108

<210>109

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>109

<210>110

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>D-Phe

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>110

<210>111

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>D-Val

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>111

<210>112

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>D-Leu

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>112

<210>113

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>D-Trp

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>113

<210>114

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>D-Thr

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>114

<210>115

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>D-Ser

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>115

<210>116

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>D-Asp

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>116

<210>117

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>D-Asn

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>117

<210>118

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>D-Glu

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>118

<210>119

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>D-Gln

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>119

<210>120

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>120

<210>121

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉4-amino-Phe

<400>121

<210>122

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉4-methoxyl group-Phe

<400>122

<210>123

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉five fluoro-Phe

<400>123

<210>124

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉2-pyridyl L-Ala

<400>124

<210>125

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉3-pyridyl Ala

<400>125

<210>126

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉4-nitro-Phe

<400>126

<210>127

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉1-naphthyl L-Ala

<400>127

<210>128

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉2-naphthyl L-Ala

<400>128

<210>129

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>2-MePhe

<400>129

<210>130

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>3-MePhe

<400>130

<210>131

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>4-MePhe

<400>131

<210>132

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉high Phe

<400>132

<210>133

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>Cha

<400>133

<210>134

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>PheNHAc

<400>134

<210>135

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>135

<210>136

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉phenyl Gly

<400>136

<210>137

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<223>Tic

<400>137

<210>138

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(8)..(8)

<223>2MePhe

<400>138

<210>139

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(7)..(7)

<223>2-Cl-Phe

<400>139

<210>140

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(7)..(7)

<223>3-Cl-Phe

<400>140

<210>141

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(7)..(7)

<223>4-C1-Phe

<400>141

<210>142

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(7)..(7)

<223>3，3-Di-Phe

<400>142

<210>143

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(7)..(7)

<223>4，4-Bi-Phe

<400>143

<210>144

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(7)..(7)

＜223〉the 4-tertiary butyl-Phe

<400>144

<210>145

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(7)..(7)

＜223〉β methyl Phe

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>145

<210>146

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>146

<210>147

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>147

<210>148

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(12)..(12)

＜223〉4-amino-Phe

<400>148

<210>149

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(12)..(12)

＜223〉4-methoxyl group Phe

<400>149

<210>150

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(12)..(12)

＜223〉five fluorine Phe

<400>150

<210>151

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(12)..(12)

＜223〉2-pyridyl Ala

<400>151

<210>152

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(12)..(12)

＜223〉3-pyridyl Ala

<400>152

<210>153

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(12)..(12)

＜223〉4-nitro-Phe

<400>153

<210>154

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(12)..(12)

＜223〉2-nitro-Tyr

<400>154

<210>155

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(12)..(12)

＜223〉4-fluoro-Phe

<400>155

<210>156

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>156

<210>157

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>157

<210>158

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>158

<210>159

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>159

<210>160

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>160

<210>161

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>161

<210>162

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>162

<210>163

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<400>163

<210>164

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>Sar

<400>164

<210>165

<211>18

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>165

<210>166

<211>16

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

＜223〉variable amino acid; See the detailed embodiment that specification sheets is submitted to

<400>166

<210>167

<211>16

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<400>167

<210>168

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(7)..(7)

<223>Dab

<400>168

<210>169

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>169

<210>170

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(7)..(7)

<223>Thz

<400>170

<210>171

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(7)..(7)

<223>Dap

<400>171

<210>172

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(7)..(7)

＜223〉Dap (amidino groups)

<400>172

<210>173

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(7)..(7)

<223>(1Me)His

<400>173

<210>174

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(7)..(7)

<223>Dab

<400>174

<210>175

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

<223>NMeHis

<400>175

<210>176

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

<223>Thz

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>176

<210>177

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

＜223〉2 pyridyl Ala

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>177

<210>178

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

＜223〉3 pyridyl Ala

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>178

<210>179

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

＜223〉thienyl Ala

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>179

<210>180

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

<223>Dab

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>180

<210>181

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

<223>Orn

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>181

<210>182

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>182

<210>183

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>183

<210>184

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

＜223〉4 amidino groups Phe

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>184

<210>185

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

＜223〉the amino Phe of 4-

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>185

<210>186

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

<223>His(Me)

<400>186

<210>187

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

<223>His(Me)2

<400>187

<210>188

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

＜223〉propargyl Gly

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>188

<210>189

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

＜223〉2-pyrrolidyl Ala

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>189

<210>190

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

<223>3-PiperidyalAla

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>190

<210>191

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

＜223〉4-piperidyl Ala

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>191

<210>192

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>192

<210>193

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>193

<210>194

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

＜223〉the 4-pyrrole is than pyridine base Ala

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>194

<210>195

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

<223>Thz(Me)

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>195

<210>196

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

＜223〉triazolyl Ala

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>196

<210>197

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Ala

<400>197

<210>198

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Ala

<400>198

<210>199

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(10)

<223>D-Ala

<400>199

<210>200

<211>15

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(8)..(8)

<223>beta-Ala

<400>200

<210>201

<211>16

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Apa

<400>201

<210>202

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<400>202

<210>203

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉4-amino methyl-phenylformic acid

<400>203

<210>204

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉(3-amino methyl)-phenylformic acid

<400>204

<210>205

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉4-aminophenyl acetate

<400>205

<210>206

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉3-aminophenyl acetate

<400>206

<210>207

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉3-amino-2-oxygen-1-piperidines-acetate

<400>207

<210>208

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉3-amino-2-oxygen-1-piperidines-acetate

<400>208

<210>209

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉(3S)-3-amino-2-oxygen-1-piperidines-acetate

<400>209

<210>210

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉3-amino-N-1-carboxymethyl-2,3,4,5-tetrahydrochysene-1H-[1]-benzazepine-2-ketone

<400>210

<210>211

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

<400>211

<210>212

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉5,5-dicyclo dipeptide analogue

<400>212

<210>213

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉5,5-dicyclo dipeptide analogue

<400>213

<210>214

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉6,5-dicyclo dipeptide analogue

<400>214

<210>215

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉3 (S)-3-amino-2-oxygen-1-azatropylidene acetate

<400>215

<210>216

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉3 (S)-3-amino-2-oxygen-1-pyrrolidine acetic acid

<400>216

<210>217

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉(3R)-3-amino-1-carboxymethyl-Valerolactim

<220>

<221>MOD_RES

<222>(9)..(9)

<223>NMeLeu

<400>217

<210>218

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(4)..(4)

<223>NMeAla

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉(3R)-3-amino-1-carboxymethyl-Valerolactim

<220>

<221>MOD_RES

<222>(9)..(9)

<223>NMeLeu

<400>218

<210>219

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉3 (S)-3-amino-2-oxygen-1-azatropylidene acetate

<220>

<221>MOD_RES

<222>(9)..(9)

<223>NMeLeu

<400>219

<210>220

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(14)..(14)

<223>Dab

<400>220

<210>221

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>221

<210>222

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Dab

<400>222

<210>223

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>223

<210>224

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>224

<210>225

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(14)..(14)

<223>Dab

<400>225

<210>226

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(14)..(14)

<223>Dap

<400>226

<210>227

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(14)..(14)

<223>Dap

<400>227

<210>228

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>228

<210>229

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Dab

<400>229

<210>230

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(14)..(14)

<223>Orn

<400>230

<210>231

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(14)..(14)

<223>Orn

<400>231

<210>232

<211>16

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>232

<210>233

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Ala

<400>233

<210>234

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Ala

<400>234

<210>235

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>235

<210>236

<211>15

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>236

<210>237

<211>16

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>237

<210>238

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>238

<210>239

<211>15

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>239

<210>240

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Dap

<400>240

<210>241

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Dap

<400>241

<210>242

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Orn

<400>242

<210>243

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Orn

<400>243

<210>244

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>244

<210>245

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>245

<210>246

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>246

<210>247

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>247

<210>248

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(10)

<223>D-Pro

<400>248

<210>249

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>249

<210>250

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Pro

<400>250

<210>251

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>251

<210>252

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>252

<210>253

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>253

<210>254

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>254

<210>255

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>255

<210>256

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>256

<210>257

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>257

<210>258

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>258

<210>259

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>259

<210>260

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>260

<210>261

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>261

<210>262

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>262

<210>263

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>263

<210>264

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>264

<210>265

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>265

<210>266

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>266

<210>267

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>267

<210>268

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Pro

<400>268

<210>269

<211>16

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Pro

<400>269

<210>270

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(8)..(8)

<223>D-Pro

<400>270

<210>271

<211>11

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(7)..(7)

<223>D-Pro

<400>271

<210>272

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>272

<210>273

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Pro

<400>273

<210>274

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>274

<210>275

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉3-amino-2-oxygen-1-piperidines-acetate

<400>275

<210>276

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉3-amino-2-oxygen-1-piperidines-acetate

<400>276

<210>277

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(8)..(8)

<400>277

<210>278

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(8)..(8)

<400>278

<210>279

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉3-aminophenyl acetate

<400>279

<210>280

<211>16

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(9)

＜223〉3-amino-2-oxygen-1-piperidines-acetate

<400>280

<210>281

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉5,5-dicyclo dipeptide analogue

<400>281

<210>282

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉6,5-dicyclo dipeptide analogue

<400>282

<210>283

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉3 (S)-3-amino-2-oxygen-1-azatropylidene acetate

<400>283

<210>284

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>284

<210>285

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>285

<210>286

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉variable amino acid

<220>

<221>MOD_RES

<222>(9)..(9)

<223>NMeLeu

<400>286

<210>287

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(4)..(4)

<223>NMeAla

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉variable amino acid

<220>

<221>MOD_RES

<222>(9)..(9)

<223>NMeLeu

<400>287

<210>288

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>288

<210>289

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>289

<210>290

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>290

<210>291

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>291

<210>292

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(6)..(6)

<223>4GuPhe

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>292

<210>293

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>293

<210>294

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>294

<210>295

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>295

<210>296

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>296

<210>297

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>297

<210>298

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>298

<210>299

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>299

<210>300

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>300

<210>301

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<400>301

<210>302

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>302

<210>303

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(10)..(10)

<223>D-Pro

<400>303

<210>304

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(9)..(9)

<223>D-Ala

<220>

<221>MOD_RES

<222>(11)..(11)

<223>NMeLeu

<400>304

<210>305

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(4)..(4)

<223>Pen

<400>305

<210>306

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>306

<210>307

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>307

<210>308

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>308

<210>309

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>309

<210>310

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>310

<210>311

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>311

<210>312

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>312

<210>313

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>313

<210>314

<211>14

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>314

<210>315

<211>15

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>315

<210>316

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(8)..(8)

＜223〉there is amino acid in variable non-natural: see the detailed embodiment that specification sheets is submitted to

<220>

<221>MOD_RES

<222>(9)..(9)

<223>NMeLeu

<400>316

<210>317

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>317

<210>318

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>318

<210>319

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>319

<210>320

<211>17

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>320

<210>321

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<220>

<221>MOD_RES

<222>(3)..(3)

<223>Pen

<220>

<221>MOD_RES

<222>(9)..(9)

<223>Sar

<220>

<221>MOD_RES

<222>(10)..(10)

<223>NMeLeu

<400>321

<210>322

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉explanation of artificial sequence: synthetic peptide

<400>322

Claims

1. peptide, it can suppress the combination of human normal immunoglobulin (IgG) Fc part to people Fc newborn infant acceptor (FcRn), comprises sequence:

-Gly-X ₆-X ₇-X ₈-X ₉-X ₁₀-X ₁₁-

Wherein:

-X ₇Be selected from phenylalanine and phenylalanine analogues;

-X ₁₁Be selected from tyrosine and tyrosine analogue; And

Wherein the length range of this peptide is 7 to 50 amino acid, and this peptide suppresses the combination of people FcRn to human IgG.

2. the peptide of claim 1, it comprises sequence:

R ₁-Gly-X ₆-X ₇-X ₈-X ₉-X ₁₀-X ₁₁-R ₂

Wherein:

-R ₁Has general formula X ₁-X ₂-X ₃-X ₄-

Wherein

-X ₁Be selected from hydrogen, acyl group and amino acid blocking group;

-X ₃Disappearance or amino acid or amino acid analogue, its energy and X ₁₀, X ₁₂Or X ₁₃Form bridge, wherein said bridge is selected from the bridge of N-terminal to the bridge of C-terminal, side chain to skeleton, reaches the bridge of side chain to side chain;

-R ₂Has general formula-X ₁₂-X ₁₃-X ₁₄-X ₁₅

Wherein

-X ₁₂Disappearance or amino acid or their analogue;

-X ₁₃Disappearance or amino acid or their analogue;

-X ₁₄Disappearance or to be selected from amino acid, length be 2-15 amino acid whose peptide and their analogue;

And

-X ₁₅Be amino group or carboxy protective group.

3. the peptide of claim 1 comprises sequence:

Gly-X ₆-Phe-X ₈-X ₉-X ₁₀-Tyr。

4. the peptide of claim 3, wherein said peptide is linear.

5. the peptide of claim 2, wherein X ₁₀, X ₁₂Or X ₁₃In at least one be can with X ₃Form the amino acid of bridge or their analogue, wherein said bridge be selected from N-terminal to the bridge of C-terminal, side chain to skeleton bridge and side chain to the bridge of side chain.

6. the peptide of claim 5, wherein X ₃With X ₁₀, X ₁₂Or X ₁₃Form bridge.

7. the peptide of claim 6, wherein X ₃With X ₁₀Form bridge.

8. the peptide of claim 6, wherein X ₃With X ₁₂Form bridge.

9. the peptide of claim 6, wherein X ₃With X ₁₃Form bridge.

10. the peptide of claim 6 wherein forms between the amino acid of bridge and has 9 amino acid.

11. the peptide of claim 6, wherein said bridge are the bridge of N-terminal to C-terminal.

12. the peptide of claim 6, wherein said bridge are the bridge of side chain to skeleton.

13. the peptide of claim 6, wherein said bridge are the bridge of side chain to side chain.

14. the peptide of claim 13, wherein said side chain is disulphide bridges, ether bridge, thioether bridge, alkene bridge or lactam bridge to the bridge of side chain.

15. the peptide of claim 14, wherein said side chain to the bridge of side chain be following each between disulphide bridges:

-halfcystine and halfcystine;

-halfcystine and homocysteine;

-halfcystine and Trolovol;

-homocysteine and homocysteine;

-homocysteine and Trolovol; And

-Trolovol and Trolovol.

16. the peptide of claim 14, wherein this side chain to the bridge of side chain be following each between lactam bridge:

-aspartic acid and Methionin;

-aspartic acid and ornithine;

-aspartic acid and 2,4-diamino-butanoic;

-aspartic acid and 2, the 3-diaminopropionic acid;

-L-glutamic acid and Methionin;

-L-glutamic acid and ornithine;

-L-glutamic acid and 2,4-diamino-butanoic;

-L-glutamic acid and 2, the 3-diaminopropionic acid.

17. the peptide of claim 2, it comprises at least one halfcystine.

18. the peptide of claim 17, wherein this at least one halfcystine is the cysteine analogs that is selected from down group:

-homocysteine;

-D-halfcystine; With

-Trolovol.

19. the peptide of claim 1 or 2, wherein X ₈And X ₉In at least one is selected from:

-glycine;

-D-amino acid;

-α-An Jiyidingsuan; With

-sarkosine.

20. the peptide of claim 1 or 2, wherein X ₈With X ₉One time-out forms the dipeptide analogue that is selected from down group:

-Beta-alanine;

-4-aminobutyric acid;

-5-aminovaleric acid;

-3-(aminomethyl) phenylformic acid;

-4-(aminomethyl) phenylformic acid;

-3-(aminophenyl) acetate;

-4-(aminophenyl) acetate;

-3-amino-2-oxygen-1-piperidines-acetate;

-(3R)-amino-2-oxygen-1-piperidines-acetate;

-(3R)-3-amino-2-oxygen-1-azepine leather acetate;

-(3R)-3-amino-2-oxygen-1-pyrrolidine acetic acid;

-(3R)-3-amino-1-carboxymethyl-Valerolactim; With

-3-amino-N-1-carboxymethyl-2,3,4,5-tetrahydrochysene-1H-[1]-benzo-aza

-2-ketone.

21. the peptide of claim 1 or 2, wherein at least one phenylalanine is a phenylalanine analogues, and every kind of described at least one phenylalanine analogues is independently selected from:

-tryptophane;

-tyrosine;

-2-amino-benzene L-Ala;

-3-amino-benzene L-Ala;

-4-amino-benzene L-Ala;

-penta fluoro benzene L-Ala;

-2-pyridyl L-Ala;

-3-pyridyl L-Ala;

-4-oil of mirbane L-Ala;

-1-naphthyl L-Ala;

-hyperphenylalaninemia;

-phenylglycine;

-2-methylbenzene L-Ala;

-3-methylbenzene L-Ala;

-4-methylbenzene L-Ala;

-2-chlorophenylalanine;

-3-chlorophenylalanine;

-4-chlorophenylalanine;

-3, the 3-diphenyl glycine;

-4,4 '-xenyl glycine;

-4-tert.-butylbenzene L-Ala;

-Cyclohexylalanine;

-(4-ammonia ethanoyl) phenylalanine;

-L-1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid;

-D-Beta-methyl phenylalanine; With

-L-Beta-methyl phenylalanine.

22. the peptide of claim 1 or 2, wherein at least one tyrosine is the tyrosine analogue, and every kind of described at least one tyrosine analogue is independently selected from:

-phenylalanine;

-4-amino-benzene L-Ala;

-4-anisole L-Ala;

-penta fluoro benzene L-Ala;

-2-pyridyl L-Ala;

-3-pyridyl L-Ala;

-4-pyridyl L-Ala;

-4-oil of mirbane L-Ala;

-2-nitrotyrosine; With

-4-fluorophenylalanine.

23. the peptide of claim 1 or 2, wherein X ₉With X ₁₀One time-out forms the dipeptide analogue that is selected from down group:

-D, the L-FriedingerShi lactan

With

-L, the L-FriedingerShi lactan

24. the peptide of claim 1 or 2, wherein at least one Histidine is the Histidine analogue, and each described at least one Histidine analogue is independently selected from:

-2,4-diamino-butanoic;

-thiazolyl L-Ala;

-2, the 3-diaminopropionic acid;

-amidino groups L-Ala;

-2-pyridyl L-Ala;

-3-pyridyl L-Ala;

-4-pyridyl L-Ala;

-thienyl alanine;

-ornithine;

-Methionin;

-arginine;

-4-Amidinophenylalaninederivatives;

-1-Methyl histidine;

-3-Methyl histidine;

-1,3-dimethyl Histidine;

-4-amino-benzene L-Ala;

-2-pyrrolidyl L-Ala;

-3-piperidyl L-Ala; With

-4-piperidyl L-Ala.

25. the peptide of claim 2, wherein X ₂Be selected from amino acid, length is 2 or 3 amino acid whose peptides and their analogue.

26. the peptide of claim 1 or 2, wherein X ₆Be positively charged amino acid or their analogue, it is selected from Methionin, ornithine, 2,4-diamino-butanoic, 2,3-diaminopropionic acid, arginine, amidino groups L-Ala and their analogue.

27. the peptide of claim 1 or 2, wherein X ₆Be die aromatischen Aminosaeuren or its analogue, it is selected from tyrosine, tryptophane, phenylalanine and their analogue.

28. the peptide of claim 1 or 2, wherein X ₆Be positively charged die aromatischen Aminosaeuren, it is selected from Histidine, 1-Methyl histidine, 2-pyridyl L-Ala, 3-pyridyl L-Ala, 4-pyridyl L-Ala, 4-amino-benzene L-Ala, 4-Amidinophenylalaninederivatives, thiazolyl L-Ala and their analogue.

29. the peptide of claim 28, wherein X ₆Be 4-Amidinophenylalaninederivatives and their analogue.

30. the peptide of claim 1 or 2, wherein X ₆Be selected from Histidine, 3-pyridyl L-Ala, 4-pyridyl L-Ala, 4-Amidinophenylalaninederivatives and their analogue.

31. the peptide of claim 30, wherein X ₆Be selected from Histidine and their analogue,

32. the peptide of claim 1 or 2, wherein X ₁₀Be selected from neutral and hydrophobic amino acid and their analogue.

33. the peptide of claim 1 or 2, wherein said peptide is a polymer.

34. the peptide of claim 33, wherein said peptide are dimer, tripolymer or the tetramer.

35. the peptide of claim 34, wherein said peptide is a dimer.

36. a peptide, it can suppress the combination of human normal immunoglobulin (IgG) Fc part to people Fc newborn infant acceptor (FcRn), comprises:

A) be selected from down the aminoacid sequence of organizing:

QRFCTGHFGGLYPCNGP(SEQ?ID?NO：1)，

GGGCVTGHFGGIYCNYQ(SEQ?ID?NO：2)，

KIICSPGHFGGMYCQGK(SEQ?ID?NO：3)，

PSYCIEGHIDGIYCFNA (SEQ ID NO:4) and

NSFCRGRPGHFGGCYLF(SEQ?ID?NO：5)；

B) with a) substantially the same aminoacid sequence;

C) with a) at least 80% identical aminoacid sequence;

D) have one, two, three, four or five than a) deletion, substitute or add the aminoacid sequence of sudden change;

E) by at least one amino acid whose aminoacid sequence of modifying a) that substitutes, wherein described at least one amino acid is substituted with naturally occurring amino acid;

F), wherein described at least one amino acid is substituted with D-amino acid and their analogue by at least one amino acid whose aminoacid sequence of modifying a) that substitutes;

G), wherein described at least one amino acid is substituted with the N-amino acid that methylates by at least one amino acid whose aminoacid sequence of modifying a) that substitutes;

H) by at least one amino acid whose aminoacid sequence of modifying a) that substitutes, wherein said at least one amino acid substitutes with the amino acid that non-natural exists; With

I) by at least one amino acid whose aminoacid sequence of modifying a) that substitutes, wherein described at least one amino acid is substituted with the amino acid analog thing,

Wherein said Toplink suppresses the combination of people FcRn to human IgG.

37. the peptide of claim 36, wherein c) in aminoacid sequence with a) at least 90% identical.

38. a dimer, it comprises the peptide of claim 36.

39. the dimer of claim 35 or 38, wherein said dimer are the products of standard reductive alkylation.

40. the dimer of claim 35 or 38, wherein said dimer are the products that reacts between single peptide monomer and the multivalence joint.

41. the dimer of claim 40, wherein said multivalence joint is selected from the joint of thiolic acid, alkohol and amine.

42. the peptide of claim 36, wherein said peptide is a tripolymer.

43. each peptide in the claim 1,2 or 36, wherein said energy peptide specific be bonded to people FcRn.

44. the peptide of claim 43, wherein said peptide is that 50fM is to 1mM to the avidity scope of people FcRn.

45. the peptide of claim 43, wherein said peptide is that 500fM is to 100 μ M to the avidity scope of people FcRn.

46. the peptide of claim 43, wherein said peptide is that 5pM is to 1 μ M to the avidity scope of people FcRn.

47. each peptide in the claim 1,2 or 36, wherein said Toplink suppress the combination of people FcRn to human IgG, and to have scope be the IC of 50fM to 1mM ₅₀

48. a peptide, it comprises following structure (SEQ ID NO:319)

49. a fusions, it comprises in the claim 1,2 or 36 each peptide and another molecule.

50. the fusions of claim 49, wherein said molecule are selected from polyoxyethylene glycol (PEG), white protein, transferrin and immunoglobulin Fc part.

51. the fusions of claim 50, wherein said molecule are the immunoglobulin Fc parts.

52. the fusions of claim 51, wherein said immunoglobulin (Ig) is selected from IgG1, IgG2, IgG3 and IgG4.

53. a medicinal compositions, it comprises in the claim 1,2 or 36 for the treatment of significant quantity each peptide.

54. the composition of claim 53, wherein preceding with using this peptide treatment human IgG serum-concentration is compared, and the peptide of treatment significant quantity can reduce the human IgG serum-concentration.

55. the composition of claim 54, wherein the human IgG serum-concentration reduces at least 5%.

56. the composition of claim 55, wherein the human IgG serum-concentration reduces at least 15%.

57. the composition of claim 56, wherein the human IgG serum-concentration reduces at least 25%.

58. a method of regulating the state of an illness comprises cell is contacted with each peptide in the claim 1,2 or 36 for the treatment of significant quantity.

59. the method for claim 58, wherein said cell expressing FcRn.

60. the method for claim 58 further comprises by regulation and control IgG serum-concentration and regulates the state of an illness.

61. the method for claim 60, wherein said IgG is specific to therapeutic protein.

62. the method for claim 61, wherein said therapeutic protein is an erythropoietin.

63. the method for claim 60, wherein said IgG is specific to gene therapy vector.

64. the method for claim 58, the wherein said state of an illness is selected from inflammatory diseases, autoimmune disorder and cancer.

65. the method for claim 64, wherein said autoimmune disorder is selected from alopecia areata, ankylosing spondylitis, antiphospholipid syndrome, the autoimmunity Addison's disease, autoimmune hemolytic anemia, autoimmune hepatitis, autoimmunity lymphoproliferative syndrome (ALPS), autoimmune thrombocytopenic purpura (ATP), Behcet, bullous pemphigoid, myocardosis, sprue-dermatitis herpetiformis, confirmed fatigue immune dysfunction syndromes (CFIDS), chronic inflammatory demyelination polyneuropathy, cicatricial pemphigoid, the CREST syndromes, cold agglutinin disease, Crohn disease, the De Gesishi disease, dermatomyositis, the juvenile form dermatomyositis, discoid lupus, the special property sent out mixed type cryoglobulinemia, fibromyalgia-fibromyositis, Graves' disease, Ge-Ba Er Shi disease, struma lymphomatosa, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura (ITP), IgA nephropathy, insulin-dependent diabetes, juvenile arthritis, lichen planus, lupus, Meniere, mixed connective tissue disease, multiple sclerosis, myasthenia gravis (MG), pemphigus, pemphigus vulgaris, pernicious anemia, polyarteritis nodosa, polychondritis, polyglandular syndrome, polymyalgia rheumatica, PM-DM, primary agammaglobulinemia, primary biliary cirrhosis, psoriatic, Raynaud's phenomenon, conjunctivo-urethro-synovial syndrome, rheumatic fever, rheumatoid arthritis, sarcoidosis, scleroderma, siogren's syndrome, the stiff man syndrome, Takayasu arteritis, temporal arteritis/giant cell arteritis, transplant rejection, ulcerative colitis, uveitis, vasculitis, vitiligo and Wei Genashi granulomatosis.

66. the method for claim 64, wherein said autoimmune disorder are selected from bullous pemphigoid, idiopathic thrombocytopenic purpura (ITP), myasthenia gravis (MG), pemphigus and transplant rejection.

67. the method for claim 66, wherein said pemphigus is a pemphigus vulgaris.

68. the method for claim 64, the wherein said state of an illness is selected from inflammatory diseases.

69. the method for claim 68, wherein said inflammatory diseases is selected from asthma, ulcerative colitis and inflammatory bowel syndrome, transformation reactions, comprise rhinallergosis/sinusitis paranasal sinusitis, skin allergic reaction (urticaria, angioedema, atopic dermatitis), food allergy, drug allergy, insectean metamorphosis reaction, mastocytosis, sacroiliitis, it comprises osteoarthritis, rheumatoid arthritis, spondyloarthropathy.

70. the method for claim 61, wherein said therapeutic protein is a thrombin.

71. the method for claim 70, wherein said thrombin are selected from Fibrinogen, thrombogen, factor V, factor VII, Factor IX, factors IX, factor X, factor XI, plasma thromboplastin antecedent, factor XI, plasma thromboplastin antecedent I, factor XI, plasma thromboplastin antecedent II or the von Willebrand factor.

72. the method for claim 70, wherein said thrombin is a Factor IX.

73. method that detects FcRn, comprise: with each peptide-labeled in the claim 1,2 or 36, wherein said detectable marker is selected from radio isotope, has enzyme, fluorophore, chemiluminogenic compound, magnetic-particle, microsphere, nanometer spheroid, vitamin H, strepto-affinity element and the digoxin that can detect product with detectable marker.

74. the method for claim 73, wherein said detection method is included in the diagnostic kit.

75. a method that is used for synthetic peptide dimer comprises that the first step is that solid-phase peptide is synthetic, second step, the 3rd step was for discharging the peptide that connects from resin for handling solid-phase resin with the reactive polypeptide of two vicinities and the molecule that is connected the peptide of these two vicinities.

76. the method for a purifying FcRn comprises:

(a) each peptide in the claim 1,2 or 36 is fixed to solid support,

(b) solution that will contain FcRn contacts with immobilized peptide on the solid support; And

(c) by purifying FcRn that solution and described solid support branch are come.