CN1212335C

CN1212335C - ICE inhibiting peptides

Info

Publication number: CN1212335C
Application number: CN 96180489
Authority: CN
Inventors: M·穆西奥; M·英特罗纳; A·曼托瓦尼
Original assignee: Applied Research Systems ARS Holding NV
Current assignee: Serono Lab
Priority date: 1996-10-31
Filing date: 1996-10-31
Publication date: 2005-07-27
Anticipated expiration: 2016-10-31
Also published as: CN1234805A

Abstract

The present invention relates to a peptide capable of being combined with ICE and/or ICE family enzyme, which is mainly composed of the amino acid sequence SEQIDNO: 1 in a sequence table. Xaa in the sequence table is selected between Asp and Ala, and the N end and/or C end of the peptide can optionally contain an amino acid or multiple amino acids. The present invention also relates to the peptide used for preparing a medical composition having the activity in pathology. The activity is required to suppress the ICE and/or ICE family enzyme.

Description

The ICE inhibiting peptide

Invention field

The present invention relates to can be in conjunction with the peptide of ICE and/or ICE family enzyme, and this peptide mainly is made of aminoacid sequence SEQID NO:1, and Xaa wherein selects between Asp and Ala, can randomly contain one or more amino acid at the N-terminal and/or the C-terminal of this peptide.

The invention still further relates to the purposes of above-mentioned peptide, is to be used to prepare have active pharmaceutical composition on the pathology, the described active enzyme that requires to suppress ICE and/or suppress ICE family.

Background of invention

ICE (il-1 β-saccharase) is a kind of heterodimer L-Cysteine HCL Anhydrous, is purified and clones (1) recently.Il-1 β (IL-1 β) is the nonactive precursor (pIL-1 β) of a kind of 33kDa or 31kDa when synthetic; IL-1 β with complete active 17.5kDa mature form starts from Ala ¹¹⁷The place is seemingly by Asp ¹¹⁶And Ala ¹¹⁷Between processing produce.So IL-1 β precursor protein is cut into sophisticated biologically active form by ICE.

Identify the ICE activity in monocyte and THP1 cell, this enzyme is at Asp ¹¹⁶-Ala ¹¹⁷And Asp ²⁴-Gly ²⁸PIL-1 β is cut at the place, generates the product (3,4) of 17.5kDa and 28kDa respectively.The cutting action in each site depends on the aspartic acid (4,6,7) of P1 position.

Obviously, represented the effector member of apoptosis mechanism with beautiful steady rhabditida (Caenorhabditis elegans) L-Cysteine HCL Anhydrous that necrocytosis albumen ced-3 is relevant.ICE is the CED-3 autoploid of describing the earliest, and known, the overexpression inducing cell program death (8) of ICE or CED-3 in rat-1 inoblast.Further research shows that also the proteolytic enzyme of ICE family may play an important role in apoptosis mechanism.

Seemingly a kind of pIL-1 β of ICE specificity processive enzyme is not because it cuts IL-1 α and other several albumen that contain many Asp-X keys.

Interleukin I L-1 α and IL-1 β are the multiple-effect cytokines, though it is similar that their sequence rarely has, but different tissues is had many similar effects, and many human diseases pathology are had effect, especially immunne response and the inflammatory process to organism has effect (9).Two kinds of proteic molecular weight all are about 17.5kDa, at the beginning of synthetic all be molecular weight be about 31kDa than the larger precursor molecule.

IL-1 is the strong inflammatory and the pyrogen sexual cell factor, usually organism is had beneficial effect, but also has extreme pathogenic effects.For example, they may participate in the pathogenesis of autoimmunization pathology (resembling systemic lupus erythematosus) symptom, particularly, they as amboceptor with cause that tissue injury is relevant, as in rheumatoid arthritis.

Many biological actions of IL-1 are similar with the effect seen in septicemia.Nearest studies show that giving IL-1 with 1 to 10ng/kg dosage vein causes fever, drowsiness, apocleisis, general myalgia, arthrodynia and headache.

Because IL-1 has multiple-effect biological activity (wherein many organism is had disadvantageous effect),, the strong effect of IL-1 controls so should being subjected to strict physiology.

The synthetic inhibition that is subjected to anti-inflammatory cytokines, prostaglandin(PG) and glucocorticosteroid of IL-1, and, exist the IL-1 of multiple level to suppress to point out to be necessary this amboceptor is carried out strict control.

Two class IL-1 acceptors are arranged, be called IL-lRI and IL-lRII.IL-lRII is non-signal IL-1 binding molecule, IL-1 is played the temptation target effect of being regulated (10-12).

At present a kind of antagonism polypeptides of IL-1 acceptor is existing reports: the 3rd member of hitherto known this receptor binding proteins family be the IL-1 acceptor antagonist (IL-lra) (13-15).Three members (IL-1 α, IL-1 β, IL-lra) discern and in conjunction with the same acceptor (IL-1R) on the cell surface; IL-1 α, IL-1 β send a signal with combining of IL-1R, and IL-lra is not then.

IL-lra is a peptide species, and it is in conjunction with IL-lRI, and with lower affinity in conjunction with IL-lRII, and without any agonist activity.IL-lra (comprises mononuclear phagocyte, polymorphonuclear leukocyte (PMN) and inoblast at different cell types.) in induce and produce through IgG, cytokine and bacterial product.

Identified and cloned two kinds of molecular form of IL-lra so far: (1) secretion property IL-lra (sIL-lra), contain one 25 amino acid whose classical leader sequence, produce 152 amino acid whose maturation proteins thus; (2) IL-lra (icIL-lra) in the born of the same parents does not have leader sequence, is retained in the born of the same parents so can infer this albumen.

SIL-lra and icIL-lra are produced by same gene.The icIL-lra transcription product is since a variable initiation site, and becomes to be arranged in the inside acceptor splicing site of sIL-lra first exon from the first variable exon montage.So two kinds of proteinic differences of Tui Ceing only are their NH in view of the above ₂End, sIL-lra preceding 21 amino acid herein at icIL-lra by 4 aminoacid replacement.

The expression of the transcription product of coding sIL-lra and icIL-lra is subjected to different regulation and control.The biological significance of icIL-lra it be unclear that.

Consider that IL-1 is relevant with the pathogeny of numerous disease, obviously need obtain the pathogenic effects that some medicines are used for limiting IL-1.

Identified and cloned icIL-lra new molecular form (16 and PCT/EP95/04023) recently.Inserting a new 63bp exon between first and second exons of this molecule in icIL-lra particular form framework produces.This new transcription product has found in inoblast, keratinocyte, activated monocyte and polymorphonuclear cell expression is arranged.The intracellular expression of COS shows that this new molecule great majority are positioned at born of the same parents, and the molecular weight of measuring through SDS-PAGE is about 25kDa.This new molecule is called II type icIL-lra (icIL-lraII).Consider that icIL-lraII is the intracellular protein the same with ICE, the applicant has also tested icIL-lraII and has suppressed the active ability of ICE.Reported the result of test among the application's the embodiment, shown that icIL-lraII suppresses the activity of ICE.

Explanation of the present invention

Main purpose of the present invention provides a kind of new peptide, and it can block the generation of IL-1 'beta ' activity form thus in conjunction with ICE, and/or more generally, can block this zymoid activity thus in conjunction with the enzyme of ICE family.So the present invention relates to can be in conjunction with the peptide of ICE and/or ICE family enzyme, it mainly is made of aminoacid sequence SEQ ID NO:1, and shown in SEQ ID NO:2 or SEQ ID NO:3 were concrete, Xaa wherein selected between Asp and Ala.Optional is that this peptide also contains one or more amino acid at its N-terminal and/or C-terminal.So the length of peptide of the present invention can be 19 to 40 amino acid, 19 to 25 better.

Specifically, according to one embodiment of the invention, described peptide mainly is made of aminoacid sequence SEQ IDNO:4 or 5.

The indefiniteness list of the L-Cysteine HCL Anhydrous of ICE family comprises: CED-3 (17), Nedd-2/ICH-1 (18,19), and Yama/CPP-32/Apopain (20,21,22), Tx/ICH-2/rel-II (23,24,25), ICErel-III (25), Mch-2 (26), ICE-LAP3/Mch-3/CMH-1 (27,28,29), ICE-LAP-6 (30) and FLICE/MACH (31,32).

Another object of the present invention provides this peptide of enough respective pure form, to be suitable as the active ingredient in the pharmaceutical composition, is used for the disease pathology that needs suppress ICE and/or suppress ICE family enzyme.

The disease pathology example that is fit to prevent, treat or diagnose with new antagonist of the present invention is lethality bacterium or virus infection, and autoimmunization and inflammatory diseases.Concrete example comprises: rheumatoid arthritis, septic shock, acute myelogenous monocytic leukemia, graft versus host immunological response, acquired immune deficiency syndrome (AIDS) (AIDS), ulcerative colitis and multiple sclerosis.

Below explanation will show other purpose of the present invention and advantage.

One embodiment of the invention are to give the patient the pharmaceutically peptide of the present invention of live vol, and described patient is in the initiation potential, need to suppress ICE and/or suppress ICE family enzyme, and perhaps the patient has shown this class pathological symptom.

The various route of administration compatible with main active ingredient all can be used, and still especially good is administered parenterally, because can obtain systemic effect at short notice like this.Therefore, administering mode is before facing operation preferably, in the operation or operation posterior vein injection drug group.The peptide dosage that gives depends on the reactive medicine prescription of being done according to patient age, body weight and individual.

Dosage can be 0.05 to 30mg/kg body weight, and dosage is 0.1 to 10mg/kg body weight preferably,

The preparation of pharmaceutical compositions that parenteral can be used become injectable forms, wherein contains main active ingredient and suitable medium.The medium that is used for administered parenterally is well-known in the art, for example water, salt solution, Lin Ge (Ringer) solution and glucose.Can contain a small amount of vehicle in the medium, to keep the stability and the perviousness of solution.

Can prepare above-mentioned solution according to general fashion, be preferably, this peptide content is between lmg/ml and 10mg/ml.

Abovely describe the present invention with reference to specific embodiment, but the content letter of this specification sheets has covered various modifications and replacement that those skilled in the art may carry out, these modifications and replace connotation and the aim do not exceed claim of the present invention and limited.

Below will the present invention will be described by embodiment, these embodiment should not understood become limitation of the invention.Embodiment will be reference with the following drawings.

Description of drawings

Fig. 1: the Northern that ICE mRNA expresses in the COS transfectional cell analyzes.With analog carrier or with the cDNA rotaring redyeing COS cell of coding people ICE, extract total mRNA, it is carried out Northern analyze, with the expression of proof ICE specific mrna.Specifically, swimming lane 1: simulation; Swimming lane 2:ICE.

Fig. 2: Western engram analysis.As preparation monocyte lysate as described in the document (4), pIL-1 β is hatched therewith; 37 ℃ hatch 60 minutes after, mixture is carried out SDS-PAGE, have precursor I 1-1 β or ripe I1-1 β by Western engram analysis proof.Specifically,

Swimming lane 1:pIL-1;

Swimming lane 2:pIL-1+ monocyte lysate.

Fig. 3: the aminoacid sequence of the peptide of being studied.Peptide A (SEQ ID NO:6) and peptide C (SEQ ID NO:4) are according to the icIL-lraII sequences Design.Peptide X (SEQ ID NO:7) is the peptide of picked at random.Peptide S (SEQ IDNO:5) only is that with the difference of peptide C two Ala have replaced two Asp residues.Peptide B (SEQ ID NO:8) is a known ICE inhibitor (Ia).

Fig. 4: the peptide of the present invention of lower concentration is to the active inhibition of ICE.Be with or without in the presence of the peptide (0.25 or 2.5 μ M), pIL-1 β (5ng) is hatched with ICE, and proof has the existence of precursor or mature form IL-1 β as described in Figure 2.Specifically,

Swimming lane 1:pIL-1;

Swimming lane 2:pIL-1+ peptide A;

Swimming lane 3:pIL-1+ peptide B;

Swimming lane 4:pIL-1+ peptide C;

Swimming lane 5:pIL-1+ICE;

Swimming lane 6:pIL-1+ICE+ peptide A (2.5 μ M);

Swimming lane 7:pIL-1+ICE+ peptide A (0.25 μ M);

Swimming lane 8:pIL-1+ICE+ peptide B (2.5 μ M);

Swimming lane 9:pIL-1+ICE+ peptide B (0.25 μ M);

Swimming lane 1O:pIL-1+ICE+ peptide C (2.5 μ M);

Swimming lane 11:pIL-1+ICE+ peptide C (0.25 μ M);

Fig. 5: the peptide of the present invention of high density is to the active inhibition of ICE.Be with or without in the presence of the peptide (40 or 400 μ M), pIL-1 β (5ng) is hatched with ICE, and proof has the existence of precursor or mature form IL-1 β as described in Figure 2.Specifically,

Swimming lane 1:pIL-1+ peptide A;

Swimming lane 2:pIL-1+ peptide B;

Swimming lane 3:pIL-1+ peptide C;

Swimming lane 4:pIL-1+ peptide X;

Swimming lane 5:pIL-1;

Swimming lane 6:pIL-1+ICE;

Swimming lane 7:pIL-1+ICE+ peptide A (400 μ M);

Swimming lane 8:pIL-1+ICE+ peptide B (400 μ M);

Swimming lane 9:pIL-1+ICE+ peptide C (400 μ M);

Swimming lane 10:pIL-1+ICE+ peptide X (400 μ M);

Swimming lane 11:pIL-1+ICE+ peptide A (40 μ M);

Swimming lane 12:pIL-1+ICE+ peptide B (40 μ M);

Swimming lane 13:pIL-1+ICE+ peptide C (40 μ M);

Swimming lane 14:pIL-1+ICE+ peptide X (40 μ M);

Fig. 6: peptide C and S are to the active inhibition of ICE.Be with or without peptide (100,300 or 1,000 μ M) existence down, pIL-1 β (5ng) is hatched with ICE, and proof has the existence of precursor or mature form IL-1 β as described in Figure 2.Specifically,

Swimming lane 1:pIL-1;

Swimming lane 2:pIL-1+ peptide C;

Swimming lane 3:pIL-1+ peptide S;

Swimming lane 4:pIL-1+ peptide B;

Swimming lane 5:pIL-1+ICE;

Swimming lane 6:pIL-1+ICE+ peptide C (0.1mM);

Swimming lane 7:pIL-1+ICE+ peptide C (0.3mM);

Swimming lane 8:pIL-1+ICE+ peptide C (1mM);

Swimming lane 9:pIL-1+ICE+ peptide S (0.1mM);

Swimming lane 10:pIL-1+ICE+ peptide S (0.3mM);

Swimming lane 11:pIL-1+ICE+ peptide S (1mM);

Swimming lane 12:pIL-1+ICE+ peptide B (1mM);

Embodiment

Material and method

Reagent

Following commercial reagent is used for cell cultures and separates:

Ficoll (Seramed, Berlin, Germany), Percoll (Pharmacia, Uppsala, Sweden), RPMI1640 (Seramed, Berlin, Germany), FCS (Hyclone Laboratories, Logan, UK), glutamine (Seramed, Berlin, Germany), with Hepes Merk, Darmastadt, Germany)

Cell

By the Ficoll gradient centrifugation, from healthy blood donor's peripheral blood acquisition mononuclearcell.Under the room temperature, 2,000rpm Percoll gradient centrifugation 30 minutes is isolated the monocyte (10) of purifying from mononuclearcell.The COS-7 cell (available from ATCC, Rockville, MD USA) is cultured in RPMI1640+10%FCS-2mM glutamine+20mM Hepes with monocyte.From Cistron Biotechnology, Pinebroon, NJ, USA obtain people's IL-1 β precursor of recombinating.In this prepared in laboratory the polyclonal antibody that all reacts with ripe and precursor forms IL-1 β.

PCR

According to open sequence (8a), obtain ICE cDNA by RT-PCR.

Reference (16) carries out 30 and takes turns RT-PCR, 95 ℃ 1 minute 30 seconds in addition, 55 ℃ 1 minute; In addition 30 seconds and 72 ℃ 1 minute other 30 seconds.Obtain oligonucleotide from Duotech (Milan Italy).The sequence that is used for the oligonucleotide of selective amplification ICE is:

" forward " ICE1:5 '-AAAAGCCATGGCCGACAAGGTC-3 ' (SEQ ID NO:9)

" oppositely " ICE2:5 '-TCTCTTCACCCTGCCCACAGAC-3 ' (SEQ ID NO:10)

The result

The expression of reorganization ICE enzyme in the COS cell

With pcr amplification the cDNA of coding people ICE.Sequence is confirmed, and with this cDNA subclone in the pSG5 expression vector.With empty carrier (simulation) or contain the carrier rotaring redyeing COS cell of ICE cDNA, after 48 hours, the expression of ICE specific mrna in the analysis of cells.As shown in Figure 3, use the high-caliber ICE mRNA of COS cell expressing of ICE cDNA transfection.

Monocyte lysate as prior art (11) preparation also can change into IL-1 β mature form (Fig. 2).So, be used as the source of ICE enzymic activity in person monocytic cell's experiment afterwards that reorganization ICE or fresh separated are obtained.

The active inhibition of ICE

The active source of preparation ICE is by testing the ability of its cutting pIL-1 β in 1 hour 37 ℃ of incubation reaction.Then mixture is carried out SDS-PAGE, prove the existence of pIL-1 β or its mature form by the Western blotting.

We design and have synthesized 4 different peptides, and have tested them and suppressed the active ability of ICE.These peptides are presented among Fig. 3, are to utilize Applied Biosystems (Foster City, solid phase synthetic instrument acquisition CA).Utilize high pressure liquid chromatography (HPLC) to verify the purity of these peptides.

(Switzerland), promptly known ICE inhibitor (Ia) is used as positive control to tetrapeptide B shown in Figure 3 for Bachem, Bubendorf.

Reference

1.a) Thomberry etc., " A novel; Cysteine protease is required for imterleukin-1 β processing in monocytes "; the processing of il-1 β needs a kind of new L-Cysteine HCL Anhydrous in the monocyte, nature (Nature), 356; 768-774,1990;

B) Ceretri etc., the molecular cloning of il-1 'beta ' converting emzyme (Molecular cloning of theinterleukin-1 β converting enzyme), science (Science), 256,97-100,1992;

2.Cameron etc., the amino acid sequence analysis of people IL-1.The evidence (Amino acid sequence analysis of human IL-1.Evidence for biochemically distinctforms of IL-1) that the IL-1 biological chemistry is multi-form, The Journal of Experimental Medicine (J.Exp.Med.) 162,790-801,1985;

3.Mosley etc., interleukin-1 receptor is in conjunction with human interleukin-1 α precursor debond il-1 β precursor but (The interleukin-l receptor binds the human interleukin-l α precursor but not theinterleukin-1 β precursor), journal of biological chemistry (J.Biol.Chem.), 262.2941-2944.1987;

4.Kostura etc., the active evaluation of monocyte specificity precursor il-1 'beta ' converting emzyme (Identification of a monocyte specific pre-interleukin-l β convertase activity), institute of American Academy of Sciences newspaper (Proc.Natl.Acad.Sci.USA), 86,5227-5231.1989;

5.Black etc., coinduction proteolytic enzyme to the activation (Activation of interleukin-1 β by a co-induced protease) of il-1 β, FEBS Lett., 427,386-390,1989;

6.Howard etc., in order to process IL-1 β precursors in two different sites, the IL-1 saccharase needs aspartic acid, and it does not cut the IL-1 α (IL-1 converting enzyme requires aspartic acidfor processing of the IL-1 β precursor at two distinct sites and dose not cleave 31kDaIL-1a) of 31kDa, Journal of Immunology (J.Immol.) 147.2964-2969,1991;

7.Griffin etc., III Int.J.Mass.Spectrom.Ion.Phys., 11,131-149,1991;

8.Miura etc., use the interleukin-11 'beta ' converting emzyme, the Mammals autoploid of C.elegans cell death gene ced-3, (the Induction of apoptosis in fibroblastsby interleukin-1 β-comverting emzyme of inductive apoptosis in inoblast, a mammalian homologue of the C.elegans celldeath gene ced-3), cell (Cell), 75,653-660,1993;

9.Dinarello, il-1 and il-1 antagonist (Interleukin-1 ans interleukin-1antagonism), blood (Blood), 77,1627-1652,1991;

1O.Colotta etc., II type interleukin-1 receptor: the temptation target (Interliekin-1type II receptor:a decoy target for IL-1 that is regulated by IL-4) that is subjected to the IL-1 of IL-4 regulation and control, science (Scieuce), 261,472-475.1993;

11.Sims etc., only by the il-1 signal (Iinterleukin-1 signallingoccurs exclusively via the type I receptor) of I receptor generation, institute of American Academy of Sciences reports, and 90,6155-6159,1993;

12.Cootta etc., immunology will today (Immunol Today), 15,562-566,1994;

13.Hannum etc., the interleukin-1 receptor antagonist activity of a kind of human interleukin-1 inhibitor (Interleukin-l receptor antagonist activity of a human interleukin-1 inhibitor), nature, 343,336-340,1990;

14.Eisenberg etc., the primary structure of human interleukin-1 receptor antagonist and the functional expression of complementary DNA (Primary sructure and functional expression form complementary DNA of a humaninterleukin-1 receptor antagonist), nature, 343,341-346,1990;

15.Carter etc., the proteic purifying of interleukin-1 receptor antagonist, clone, expression and biological characteristics are described (Purification, cloning, expression and biological characterisation of an interleukin-1receptor antagonist protein), nature, 344,633-638,1990;

16.Muzio etc., the clone of a kind of new il-1 (IL-1) receptor antagonist (IL lra) allotype body and evaluation (Cloning and characterisation of a new isoform of interleuk9in-1 (IL-1) receptorantagonist:(IL lra)), The Journal of Experimental Medicine (J Exp Med), 182632,1995;

17.Yuan etc., the C.elegans cell death gene ced-3 a kind of albumen similar (C.elegans cell death gene ced-3 encodes a protein similar tomammalian interleukin-1 β-converting enzyme) of encoding to Mammals il-1 β-saccharase, cell, 75,641-652,1993;

18.Kumar etc., utilize the gene induced apoptosis of mouse nedd2, albumen (the Induction of apoptosis by the mouse nedd2 gene that this genes encoding is similar with Mammals il-1 β-saccharase to C.elegans cell death gene ced-3 product, which encodes a protein similar tothe product of the C.elegans cell death gene ced-3 and the mammalian interleukin-1 β-converting enzyme), genetics progress (Genes Dev.), 8,1613-1626,1994;

19.Wang etc., Ich-1, a kind of Ice/ced-3 genes involved, the positive modulator of Codocyte program death and negative regulation thing (Ich-1, an Ice/ced-3-related gene, encodes both positive and negativeregulators of programmed cell death), cell, 78,739-750,1994;

20.Fernandes-Alnemri etc., CPP-32, a kind of new people's apoptosis albumen, has homology (CPP-32 with C.elegans necrocytosis PROTEIN C ED-3 and Mammals il-1 β-saccharase, anovel human apoptotic protein with homology to C.delegans cell death protin CDE-3and mammalian interleukin-1 β-converting enzyme), journal of biological chemistry (J.Biol.Chem.), 269,30761-30764,1994;

21.Nicholson etc., to the evaluation and the inhibition (Identification and inhibition of ICE/CED-3 protease necessary for mammalianapoptosis) of the essential ICE/CED-3 proteolytic enzyme of mammalian cell program death, nature, 376,37-43,1995;

22.Tewari etc., Yama/CPP-32 β, a kind of Mammals autoploid of CED-3, be to cut dead matrix poly-(ADP-ribose) but the CmA-arrestin enzyme (Yama/CPP-32 β, a mammalianhomologue of CED-3, is a CmA-inhibitable protease that cleaves the death substratepoly (ADP-ribose) polymerase) of polysaccharase, cell, 81,801-809,1995;

23.Faucheu etc., a kind of new human protease similar inducing cell program death in transfectional cell (A novel human protease similar to the interleukin-1 β-convertingenzyme induces apoptosis in transfected cells (EMBO J.) to il-1 β-saccharase, 14,1914-1922,1995;

24.Kamens etc., the evaluation of ICH-2 and characteristic description, the newcomer of L-Cysteine HCL Anhydrous il-1 β-saccharase family (Identification and characterisation of ICH-2, a novel member of theinterleukin-1 β-converting enzyme family of cysteine proteases), journal of biological chemistry, 270,15250-15256,1995;

25.Munday etc., the molecular cloning of ICE rel-II and ICE rel-III and its apoptosis activity, the member of L-Cysteine HCL Anhydrous ICE/CED-3 family (Molecular cloning and proapoptoticactivity of ICE rel-II and ICE rel-III members of the ICE/CED-3 family of cysteineproteases), journal of biological chemistry, 270,15870-15876,1995;

26.Fernandes-Alnemri etc., Mch-2, newcomer (the Mch-2 of L-Cysteine HCL Anhydrous apoptosis CED-3/ICE group family, a new member of the apoptotic CED-3/ICE cysteineprotoease gene family), cancer research, 55,2737-2742.1995;

27.Duan etc., a kind of new Mammals autoploid of C.elegans necrocytosis PROTEIN C ED-3, ICE-LAP3 is activated (ICE-LAP3 in Fas-and TNF inductive apoptosis, a novelmammalian homologue of the C.elegans cell death protein CED-3, is activated duringFas-and TNF-induced apoptosis), journal of biological chemistry, 2771,35013-35035,1996;

28.Fernandes-AInemri etc., Mch-3, a kind of new people's apoptosis L-Cysteine HCL Anhydrous (Mch-3 with the CPP-32 height correlation, a novel human apoptotic cysteine protease highlyrelated to CPP-32), cancer research (Cancer Res), 55,6045-6052,1995;

29.Lippke etc., the evaluation of CPP-32/Mch-2 autoploid and characteristic description, similar new L-Cysteine HCL Anhydrous (the Identification and characterisation of CPP-32/Mch-2homologue 1 of a kind of and CPP-32, a novel cysteine protease similar to CPP-32), journal of biological chemistry, 271,1825-1828,1996;

30.Duan etc., the newcomer of CED-3/ICE gene family, ICE-LAP6 is activated (ICE-LAP6 by cytotoxic T cell proteolytic enzyme granzyme B, a novel member of the CED-3/ICE gene family, isactivated by the cytotoxic T cell protease granzyme B) journal of biological chemistry was published in 1996;

31.Muzio etc., FLICE, a kind of new FADD homology ICE/CED-3 sample proteolytic enzyme, it is the new dead inducibility signal of CD95 (Fas/Apo-1) complex body (FLICE, a novel FADD-homologousICE/CED-3-like protease, is recruited to the CD95 (Fas/Apo-1) Death-InducingSignalling Complex), cell, 85,817-827,1996;

32.Boldin etc., a kind of new MORT-1/FADD-interaction protein enzyme, MACH relates to necrocytosis (the Involvement of MACH of Fas/Apo-1 and TNF receptor-inducible, a novel MORT-1/FADD-interacting proteases, in Fas/Apo-1 and TNF receptor-induced cell death), cell, 85,803-815;

Sequence table

(1) generally detailed

(i) applicant:

(A) title: APPLED RESEARCH SYSTEMS ARS HOLDING N.V.

(B) street: 14, JOHN B.GORSIRAWEG

(C) city: CURACAO

(E) country: THE NETHERLAND S ANTILLES

(F) postcode:

(G) phone: 599-9-639300

(H) fax: 599-9-614129

(ii) denomination of invention: ICE INHIBTING PEPTIDES

(iii) sequence quantity: 10

(iv) computer-reader form:

(A) media type: floppy disk

(B) computer: IBM PC compatible

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

(D) software: PatentIn Release#1.0, Version#1.30 (EPO)

(2) information of SEQ ID NO:1:

(i) sequence signature:

(A) length: 19 amino acid

(B) type: amino acid

(C) chain:

(D) topological framework: linearity

(ii) molecule type: peptide

(ix) feature:

(A) title/code name: decorating site

(B) position: 2

(D) out of Memory :/annotating: Xaa is to be good amino acid with Asp or Ala, is that Asp is better

(ix) feature

(A) title/code name: decorating site

(B) position: 19

(xi) sequence description: SEQ ID NO:1:

Ala?Xaa?Leu?Tyr?Glu?Glu?Gly?Gly?Gly?Gly?Gly?Gly?Glu?Gly?Glu?Asp

1 5 10 15

Asn?Ala?Xaa

(2) information of SEQ ID NO:2:

(i) sequence signature:

(A) length: 19 amino acid

(B) type: amino acid

(C) chain:

(D) topological framework: linearity

(ii) molecule type: peptide

(xi) sequence description: SEQ ID NO:2:

Ala?Asp?Leu?Tyr?Glu?Glu?Gly?Gly?Gly?Gly?Gly?Gly?Glu?Gly?Glu?Asp

1 5 10 15

Asn?Ala?Asp

(2) information of SEQ ID NO:3:

(i) sequence signature:

(A) length: 19 amino acid

(B) type: amino acid

(C) chain:

(D) topological framework: linearity

(ii) molecule type: peptide

(xi) sequence description: SEQ ID NO:3:

Ala?Ala?Leu?Tyr?Glu?Glu?Gly?Gly?Gly?Gly?Gly?Gly?Glu?Gly?Glu?Asp

1 5 l0 15

Asn?Ala Ala

(2) information of SEQ ID NO:4:

(i) sequence signature:

(A) length: 24 amino acid

(B) type: amino acid

(C) chain:

(D) topological framework: linearity

(ii) molecule type: peptide

(xi) sequence description: SEQ ID NO:4:

Met?Ala?Leu?Ala?Asp?Leu?Tyr?Glu?Glu?Gly?Gly?Gly?Gly?Gly?Gly?Glu

1 5 10 15

Gly?Glu?Asp?Asn?Ala?Asp?Ser?Lys

20

(2) information of SEQ ID NO:5:

(i) sequence signature:

(A) length: 24 amino acid

(B) type: amino acid

(C) chain:

(D) topological framework: linearity

(ii) molecule type: peptide

(xi) sequence description: SEQ ID NO:5:

Met?Ale?Leu?Ala?Ala?Leu?Tyr?Glu?Glu?Gly?Gly?Gly?Gly?Gly?Gly?Glu

1 5 10 15

Gly?Glu?Asp?Asn?Ala?Ala?Ser?Lys

20

(2) information of SEQ ID NO:6:

(i) sequence signature:

(A) length: nine amino acid

(B) type: amino acid

(C) chain:

(D) topological framework: linearity

(ii) molecule type: peptide

(xi) sequence description: SEQ ID NO:6:

Glu?Gly?Glu?Asp?Asn?Ala?Asp?Ser?Lys

1 5

(2) information of SEQ ID NO:7:

(i) sequence signature:

(A) length: 13 amino acid

(B) type: amino acid

(C) chain:

(D) topological framework: linearity

(ii) molecule type: peptide

(xi) sequence description: SEQ ID NO:7:

Phe?Lys?Asp?Pro?His?Gly?Leu?Trp?Lys?Gly?Leu?Ser?His

1 5 10

(2) information of SEQ ID NO:8:

(i) sequence signature:

(A) length: six amino acid

(B) type: amino acid

(C) chain:

(D) topological framework: linearity

(ii) molecule type: peptide

(iii) suppose: non-

(iv) antisense: non-

(ix) feature:

(A) title/code name: decorating site

(B) position: 1

(D) out of Memory :/annotating: Xaa is an ethanoyl

(ix) feature

(A) title/code name: decorating site

(B) position: 6

(D) out of Memory :/annotating: Xaa is-CHO

(xi) sequence description: SEQ ID NO:8:

Xaa?Tyr?Val?Ala?Asp?Xaa

1 5

(2) information of SEQ ID NO:9:

(i) sequence signature:

(A) length: 22 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological framework: linearity

(ii) molecule type: other nucleic acid molecule

(A) illustrate :/desc represents " oligonucleotide "

(iii) suppose: non-

(iv) antisense: non-

(xi) sequence description: SEQ ID NO:9:

AAAAGCCATG?GCCGACAAGG?TC 22

(2) information of SEQ ID NO:10:

(i) sequence signature:

(A) length: 22 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topological framework: linearity

(ii) molecule type: other nucleic acid molecule

(A) illustrate :/desc represents " oligonucleotide "

(iii) suppose: non-

(iv) antisense: non-

(xi) sequence description: SEQ ID NO:10:

TCTCTTCACC?CTGCCCACAG?AC 22

Claims

One kind can be in conjunction with the peptide of il-1 β-saccharase, its aminoacid sequence is selected from SEQ ID NO:4 or SEQ ID NO:5.
2. the described peptide of claim 1 is used for making treatment autoimmune disease, the purposes of the medicine of lethality bacterium and virus infection or inflammatory diseases.
3. purposes according to claim 2, described disease is an autoimmune disease.
4. purposes according to claim 2, described disease are lethality bacterium and virus infection.
5. purposes according to claim 2, described disease is an inflammatory diseases.
6. purposes according to claim 2, described disease are selected from rheumatoid arthritis, septic shock, acute myelogenous monocytic leukemia, graft versus host immunological response, acquired immune deficiency syndrome (AIDS), ulcerative colitis and multiple sclerosis.
7. pharmaceutical composition contains the described peptide of claim 1 and one or more pharmaceutically acceptable carrier and/or vehicle.