TWI314558B - Antibodies to human il-1beta - Google Patents

Description

1314558 A7 B7 __ V. INSTRUCTION DESCRIPTION (1) The present invention relates to an antibody to human interleukin I beta (IL-i) and the use of the antibody for the treatment of IL-1 vector disease or discomfort. Interleukin-1 (IL-1) is a substrate produced by cells of the immune system and acts as a mediator of the acute phase of inflammation. Improper or over-producing IL-1 (specifically IL-1 /?) is associated with pathology of many diseases or discomforts, such as sepsis, septic or endotoxic shock, allergies, asthma, bone loss, ischemia, Stroke 'rheumatoid arthritis and other inflammatory discomfort. IL-1 Θ antibodies have been proposed for the treatment of diseases or disorders of the IL-1 vector, as discussed in WO 95/01997 and its preamble. The present invention produces a human IL-1 modified antibody for use in the treatment of a disease or discomfort of the IL-1 vector. The present invention can provide an IL-ly5 binding molecule comprising an antigen binding site comprising at least one immunoglobulin heavy chain variable region (VH), wherein the variable region comprises a CDR1, a CDR2, a CDR3 in a sequence hypervariable region. The amino acid sequence of the CDR1 is Ser-Tyr-Trp-Ile-Gly, and the amino acid sequence of the CDR2 is Ile-Ile-Tyr-Pro-Ser-Asp-Ser-Asp-Thr-Arg_Tyr-Ser-Pro -Ser-Phe-Gln-Gly, and the amino acid sequence of the CDR3 is Tyr-Thr-Asn-Trp-Asp-Ala-Phe-Asp-Ile, and its immediate counterpart. In a first aspect, the invention provides a single-region IL_1/? binding molecule comprising a separate immunoglobulin heavy chain comprising a heavy chain variable region (Vh) as defined above. In a first aspect, the invention also provides a heavy chain variable region (νΗ) and a light chain variable region 々 binding molecule, wherein the IL_1/? binding molecule comprises at least one antigen binding position, comprising: 4- This paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) (please read the notes on the back and fill out this page)

Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumer Cooperative, Printed A7 B7 1314558 V. INSTRUCTIONS (2) a) An immunoglobulin heavy chain variable region (vh)' whose variable region includes CDR1 and CDR2 , CDR3, the amino acid sequence of the CDR1 is Ser-Tyr-Trp-Ile-Gly' The amino acid sequence of the CDR2 is 116-Ile-Tyr-Pro-Ser-Asp-Ser-Asp-Thr-Arg-Tyr -Ser-Pro-Ser-Phe-Gln-Gly, and the amino acid sequence of the CDR3 is Tyr-Thr-Asn-Trp-Asp-Ala-Phe-Asp-Ile, and b) an immunoglobulin light chain The variable region (VL) comprises a CDR3' hypervariable region, the amino acid sequence of which is Gln-Gln-Arg-Ser-Asn-Trp-Met-Phe-Pro; and its immediate counterpart. A specific embodiment of the second aspect of the present invention provides an IL-1 binding molecule comprising both a heavy chain variable region (VH) and a light chain variable region (VL), wherein the IL-1/? binding molecule comprises _One less antigen binding position' includes: a) an immunoglobulin heavy chain variable region (VH), the variable region of which comprises the CDR1, CDR2, CDR3, and the amino acid sequence of the CDR1 Is Ser-Tyr-Trp-Ile-Gly, the amino acid sequence of the CDR2 is -Ile-Tyr-Pro-Ser-Asp-Ser-Asp-Thr-Arg-Tyr-Ser-Pro-Ser-Phe-Gln -Gly, and the amino acid sequence of the CDR3 is Tyr-Thr-Asn-Trp-Asp-Ala-Phe-Asp-Ile, and b) an immunoglobulin light chain variable region (VJ, variable region thereof) The sequence hypervariable region includes CDR1', CDR2', CDR3', and the amino acid sequence of the CDR1 is Arg-Ala-Ser-Gln-Ser-Val-Ser-Ser-Tyr-Leu-Ala, the CDR2' The amino acid sequence is Asp-Ala-Ser-Asn-Arg-Ala-Thr, and the amino acid sequence of the CDR3' is Gln-Gln-Arg-Ser-Asn-Trp-Met--5- Applicable to China National Standard (CNS) A4 specification (210 X 297 mm) (Please read the notes on the back first) Complete this page) -n n I · 1 · a 6 ', Ministry of Economic Affairs Intellectual Property Office employees consumer cooperatives printed 1314558 A7, Description (3 invention)

Phe-Pro; and its straight counterparts. Unless otherwise specified, the amino acid sequence of the polypeptide chain and the amino acid sequence described herein begins with Ν:^ and ends at the C-terminus. When the antigen binds to human F眭, and the πγ, &; 免 w w position also includes (VH) and (VL) k, it may be located in the same polypeptide molecule or preferably each region is located on a different chain 'VH is an immunoglobulin Heavy chain or ligature, ^ Α re-fragment <score and VL is part of the immunoglobulin light chain or a fragment thereof. IL-1々 combines " refers to any molecule that can bind to the IL-1 antigen in conjunction with other molecules. Zhuang people, ', mouth σ reaction can be displayed using standard methods (qualitative test), including material IL, bioassay of its degree of inhibition or any kind of combination test, and refer to the relevant control control test 'control test Use of money - a kind of non-phase, but the isomer

<柷 (such as anti-CD25 antibody). The binding of the IL_1/? antigen molecule of the present invention to il_M is preferably shown by a competitive binding assay. Examples of antigen-binding molecules include antibodies produced by B cells or fusion tumors, and CDR-grafted or human antibodies or any fragments thereof, such as F(ab,)2 and Fab fragments, and single-stranded or single-region antibodies. The single-region antibody comprises a variable region consisting of an antibody heavy chain and a light chain, in which a peptide crosslinker (usually composed of 10 to 30 amino acids, preferably 15 to 25 amino acids) is used. Covalent link. Therefore, the structure does not contain the heavy chain and the constant region of light, and it is believed that the antigenicity of the small peptide spacer should be lower than that of the intact constant region. &quot;Chimeric antibody&quot; refers to an antibody in which the constant regions of the heavy or light chain or both are derived from humans, while the variable regions of the heavy and light chains are derived from non-human (such as murine) or Derived from humans but from different human antibodies. 6- This paper scale applies to China National Standard (CNS) A4 specification (21〇x 297 public Γ Γ Please read the back note and then fill out this page) 订·! Λ Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative print 1314558 A7 B7 V. Inventive Note (4), CDR-transplanted antibody refers to an antibody in which the hypervariable regions (CDRs) are derived from a donor antibody (such as a non-human (such as murine) antibody or a different human antibody). And all or substantially all other parts of the immunoglobulin (such as the constant region and the high-preservation region in the variable region (ie, the framework region)) are derived from the recipient antibody (eg, from humans). However, the CDR-grafted antibody The framework region may contain some amino acid of the donor sequence, such as the portion of the framework region adjacent to the hypervariable region. "Human antibody" refers to an antibody in which the constant region of the heavy and light chains is derived from the variable region. Human, or substantially the same sequence as human origin, does not need to be from the same antibody and contains antibodies made by mice, wherein the genes of the murine immunoglobulin variable and constant regions have been mapped by humans. Substituted as a general term for EP 0 546 073 Bl, USP 5, 545, 806, USP 5, 506, 826, USP 5, 625, 126, USP 5, 334, 425, USP 5, 610, 610, USP 5, 770, 429, EP 0 438 474 B1, and EP 0 463 151 B1. A binding molecule human antibody, especially the AAL 160 antibody described in the following examples. The heavy chain and light chain variable regions in the preferred chimeric antibody are derived from humans, such as AAL 160 antibody, which is shown in Sequence identification number 1 and sequence identification number 2. The constant region preferably also includes a suitable human constant region, as described in &quot;Sequences of Proteins of Immunological interest&quot;, Kabat EA et al, US Department of Health and Human The hypervariable region can be associated with any kind of skeleton region, but is preferably derived from humans. The appropriate framework is described in Carbate EA et al., supra. The heavy chain skeleton is a human heavy chain skeleton. The paper size shown in serial identification number 1 applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm). Read the notes on the back and fill in this page) --------tT---------i. Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumption Cooperative, Printed Economy Ministry, Intellectual Property Bureau, Staff Consumer Cooperative, Printed 1314558 A7 ____B7____ V. Description of the invention (5) AAL 16〇 antibody. It is stored in the sequence of rush, FR2, FR3, and sigh 4 zones. Similarly, sequence identification number 2 shows a preferred AAL 16 〇 light chain backbone, which is stored in the FR1', FR2', FR3', FR4' regions. Accordingly, the present invention also provides an IL-10 binding molecule comprising at least one antigen binding site comprising a first region having an amino acid sequence substantially identical to that shown in Sequence Identification Number 1 (the amino acid of which begins with Position and finally position 118), or a set of first regions as described above and a second region having an amino acid sequence substantially identical to that shown in Sequence Identification Number 2 (the amino acid of which begins at position 1 and finally positions 1〇7). Monoculture antibodies raised against human natural proteins are usually formed in non-human systems, such as mice. Therefore, when administered to a human, a heterologous antibody produced by a fusion tumor can cause an inappropriate immune response, which is mainly caused by the constant region of the heterologous immunoglobulin. This significantly limits the use of this antibody because it cannot be administered chronically. Therefore, the best use of single-stranded, single-region, CDR_transplant, or especially human antibodies, does not readily cause a physical allergic reaction when administered to humans. For the above reasons, a more preferred IL-1 Θ binding molecule of the invention is selected from the group consisting of a human anti-IL-1 antibody comprising at least: a) an immunoglobulin heavy chain or a fragment thereof comprising (1) a heavy chain a variable region comprising a sequence comprising a mutated region CDR1, CDR2, CDR3 and (ii) a constant region of a human heavy chain or a fragment thereof; the amino acid sequence of the CDR1 is Ser-Tyr-Trp-Ile-Gly, an amine of the CDR2 The base acid sequence is 14-1 &amp; 1*〇-Ser-Asp-Ser-Asp-Thr-Arg-Tyr-Ser-Pro-Ser-Phe-Gln-Gly, and the amino acid sequence of the CDR3 is Tyr-Thr-Asn-Trp-Asp-Ala -Phe- -8 - This paper size is applicable to China National Standard (CNS) A4 specification (210 X 297 mm) (please read the notes on the back and fill out this page)

1314558 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed A7 V. Inventive Note (7) (1) The supervariant region CDR1, CDR2 & CDR3 as a whole at least 8〇%, preferably at least 90%, more preferably at least 95% The supervariation region shown in sequence identification number 1 is the same, and (11) it inhibits the binding of IL-1 to its acceptor, and the degree of inhibition is substantially the same as that of the framework region having the same skeleton region as molecule X, but it has The mutated region number 1 is identical to the mutated region CDR1, CDR2 and CDR3 or any IL-1 binding molecule, wherein each binding position has at least two regions (Molecule X') (1) wherein the hypervariable region CDR1, CDR2, CDR3, CDR3, and At least 80%, preferably at least 9%, more preferably at least 95% of the CDR1 and CDR2' as desired are identical to the hypervariable regions shown in Sequence Identification Numbers 1 and 2, and (11) inhibit IL -1 结合 binds to its acceptor, and its degree of inhibition is generally as a reference molecule having the same framework and constant region as X', but it has the same mutated region CDR1, CDR2 as shown in sequence identification numbers 1 and 2. , CDR3, and CDR3, and Required CDR1, and CDR2-. If at least 80% of the same amino acid residues are in similar positions and the sequence is ideally arranged, then at least 8 % of the amino acid sequences currently described are identical, and the amino acid sequence is notched or inserted as a different residue. base. Inhibition of binding of IL-I々 to its receptor is conveniently tested in a variety of assays, including those described below. The IL_1/? acceptor used is preferably a j-type acceptor. The term "degree of inhibition" refers to the statistic basis, refer to -10- (please read the notes on the back? Please fill out this page again)

Ministry of Economic Affairs, Intellectual Property Bureau, employee consumption cooperatives printed! 314558

The invention has the same IL_丨々 binding inhibition curve as one of the previous analyses. (8) The analytical assay used can be a competitive inhibition ratio. IL-1 /5 acceptor and IL_丨卢 binding molecule of the invention. Optimally, the human IL-1 cold antibody comprises at least a) a heavy chain comprising an amine having substantially the same identity as shown in SEQ ID NO: a nucleic acid sequence variable region that begins at the position of the amino acid and finally positions the 118 amino acid and the human heavy chain constant region, and b) the light chain, which comprises the amine having substantially the same amine as the sequence identification number. The variable region of the acid sequence (starting at position 丨-amino acid and finally position 107 amino acid) and the human light chain constant region. The human (4) strange region can be w r3, m, w 4 or ε-type Preferably, the human light chain binding region can be 疋 or; type I (which includes; I, 2 and λ3 subtypes) but preferably "type. The amino acid sequences of all of these constant regions are derived from Cabet et al., supra. The IL-1 reading molecules of the present invention can be prepared by DNA recombination techniques. In this case, one or more DNA molecules encoding the binding molecule must be constructed, placed under appropriate control sequences and transferred to the appropriate host organism for expression. In a very general manner, according to this, (i) a DNA molecule capable of compiling the single-region scorpion-binding molecule of the present invention, the single-stranded IL(10) molecule of the present invention, the heavy chain or light of the bismuth dimer of the present invention The chain or fragment thereof, and the mouth knife (9) use the DNA molecule of the present invention to prepare the H-knot-11 of the present invention in a recombinant manner - (please read the phonetic transcription on the back side and then fill out this page)

1314558 A7 B7 V. INSTRUCTIONS (9) Use of molecular. The current state of the art has reached a level that is familiar to those skilled in the art to synthesize DNA molecules of the invention that are compatible with the information provided herein (i.e., the amino acid sequence of the hypervariable region and its coding DNA sequence). Methods for constructing variable region genes can be found, for example, in EPA 239 400, and can be summarized as follows: A gene encoding a variable region of MAb A having any specificity is selected. A DNA fragment encoding the framework and the hypervariable region is identified and the DNA fragment encoding the hypervariable region is removed such that the DNA fragment encoding the framework region is fused to the appropriate restriction site at the junction. The restriction position can be generated by mutagenesis of the DNA molecule in place via standard procedures. A double-stranded synthetic CDR cassette, such as the sequence of Sequence Identification Number 1 or 2, was prepared via DNA synthesis. These cassettes have a sticky end so that they can be attached to the joint of the skeleton. Furthermore, it is not necessary to obtain mRNA from a productive fusion tumor cell line to obtain a DNA structure encoding the IL-1/? binding molecule of the present invention. Thus, PCT Patent Application WO 90/07861 provides a complete description of the manufacture of antibodies using recombinant DNA technology, in which only the nucleotide sequence of the gene is provided. The method involves the synthesis of a number of oligonucleotides, amplification by PCR, and splicing to obtain the desired DNA sequence. Printed by the Intellectual Property Office of the Ministry of Economic Affairs and the Consumer Cooperatives (please read the notes on the back and fill out this page). An expression vector containing a suitable promoter gene or a gene encoding a heavy bond and a light bond binding region can be obtained publicly. Therefore, when the DNA molecule of the present invention is prepared, it can be conveniently transferred to an appropriate expression carrier. DNA molecules encoding single chain antibodies can also be prepared by standard methods as described in WO 88/1649. In view of the above, it is not necessary to specify a fusion tumor or cell strain that is fully compliant with the stated standards. -12- This paper size is applicable to China National Standard (CNS) A4 specification (210 X 297 mm) 1314558 A7

The first and second dna structures are included in a particular embodiment of the invention to produce an IL_1/? binding molecule as described below: The first DNA structure can encode a heavy chain or a fragment thereof and comprise a) one can be encoded In the first part of the variable region, the variable region comprises a selective backbone and a supervariable region, the supervariable regions are the sequences CDR1, cdr2 and CDR3, and the amino acid sequence thereof is found in the sequence identification number 丨; the first part begins with a codon encoding the first amino acid of the variable region and finally - a codon encoding the variable region final amino acid, and b) a second portion encoding a heavy chain constant region or a fragment thereof, Starting with a codon that encodes the first amino acid of the heavy chain constant region and finally a codon that encodes the final amino acid of the constant region or a fragment thereof, and finally a stop codon. Preferably, the first portion encodes a variable region having an amino acid sequence substantially identical to that found in Sequence Identification Number 1, the amino acid starting at position 1 and finally at position 118. More preferably, the first portion has a nucleotide sequence as seen in Sequence Discrimination No. 1 whose nucleic acid starts at position 1 and finally at position 354. Also preferably, the second portion encodes a constant region of the human heavy chain' more preferably a constant region of the human chain. The second portion can be a DN A fragment (containing an insertion sequence) or a cdna fragment (without an insertion sequence) from the genome. The second DNA construct may encode a light chain or a fragment thereof and include a) a first portion of a variable region encoding a variable region comprising a selective backbone and a hypervariable region; the hypervariable region is a CDR3, And if necessary, CDR1' and CDR2, the amino acid sequence is found in the serial identification number 13. The paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) (please read the precautions on the back) Fill in this page) · ! i I Book! ! ! 'Ministry of Economics Intellectual Property Bureau employee consumption cooperative printed i3l4558

V. Description of invention (11 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed on - can encode variable region s-amino acid... 丄, code variable region final amino acid codon, and -) - can be coded light The second portion of the chain constant region or a fragment thereof can encode the light chain constant region first, ^Shimin-ra-like &amp; K, code and finally a knottable constant region or its fragment is finally Τ,. a. , amine Κ in the code to 'final reconnection finally preferred, can be coded variable number 9 士 foundation 4 by the younger brother a sickle has the same amino acid sequence as the sequence identification, 唬 2 big ,, position 107. More preferably, the base starts at position 1 and finally the branch of number 2 = the portion has the sequence identification as shown in the sequence identification. The nucleoside acid starts at position 1 and finally positions. :7 and ^, 罘二邵分 can encode the human light chain constant region, more preferably human; ί chain constant region. L Wen Jia Ma II = contains 1 "々 binding molecule" in which - or more _, gas 扪Π CDR1, CDR2, or CDR3, the residue is replaced by the residue found in SEQ ID NO: 2; (eg, corresponding to the position of the sequencer - mutation). The invention comprises a DNA sequence encoding the substitution -1 = sub. The invention specifically comprises an m-knot &quot;, wherein CDR1i or CDR2, one or more The residue has been replaced by a residue found in column identification number 2. In the first and second DNA structures, the first and second portions can be inserted into the sequence, and the promoter can be conveniently placed between the first and The insertion sequence between the second part. The presence of the promoter (which is transcriptional, non-translated) can help to effectively turn green. In a particular embodiment, the first and second DNA constructs comprise a heavy chain gene Promoter, which is best derived from humans. -14 - This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm)

^ · I I-------------- (Please read the notes on the back and fill out this page) 1314558 A7 B7 V. Inventions (12) (Please read the notes on the back first) Fill out this page) The control of each DNA structure is placed under the appropriate control array, and the specific strain is controlled by an appropriate promoter gene. Any kind of promoter can be used to adapt to the host organism, and the DNA structure can be transferred and expressed. However, if the expression occurs in mammalian cells, it is particularly preferred to use a promoter gene for an immunoglobulin gene, &amp; a large cell virus (CMV) promoter (such as a human CMV promoter). The desired antibody can be prepared in a cell culture or in a transgenic animal. Appropriate transgenic animals can be obtained by standard methods, including microinjection of the first and second DNA structures placed in appropriate control sequences into the egg, transfer of the prepared egg to a suitable pseudopreg hen and selection To represent the progeny of the desired antibody. When preparing antibody chains in cell culture, the 譲 structure must first be inserted

A single expressive vector or two separate but compatible expressions are preferred. T Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing According to this, the present invention also provides an expression vector which can be replicated in a prokaryotic or sputum nuclear cell strain, which comprises at least one dna junction as described above and then contains a DNA structure. Each of the expression vectors is transferred to a suitable host. When the DNA structure is inserted into two expression vectors, respectively, it can be transferred separately (ie, per-cell-type carrier) or co-transfer, the latter being preferred: the appropriate host organism can be bacteria, yeast, or lactating Animal cell lines, but the latter is preferred. More preferably, a mammalian cell line derived from lymph, such as a myeloma, a fusion tumor or a normal immortalized B_cell, is convenient for any endogenous antibody heavy or light chain. When expressed in mammalian cells, it is preferred to bind the H-binding molecule -15 1314558 A7

Printed by the Intellectual Property Office of the Ministry of Economic Affairs, Employees' Consumption Cooperatives. V. INSTRUCTIONS (13) The incorporation of the coding sequence into the host cell DNA allows or facilitates the large-scale expression of the IL 1 /5~ molecule. The criteria for identifying and selecting which cells can be used to incorporate the binding sequence of the binding molecule and placing it in a favorable position are the number of IL-1 binding molecules that can be expressed. Any suitable selectable marker can be used to prepare a host cell containing the IL-1 々 binding molecule coding sequence; for example, sinensis/aminothymidine or the same selection system can be used. Preferably, the system of the present invention incorporates a GS-based amplification/selection system, as described in EP 0256055 B, EP 0 323 997 B, and European Patent Application 89 303 964 4 . Preferably, the vector may contain other sequences which may aid in the expression, processing and export of the expressed protein; for example, the vector may comprise a leader sequence in addition to the coding sequence. In a still further aspect of the present invention, there is provided a method of preparing a specific binding molecule comprising (1) cultivating an organism transformed with an expression vector as described above and (ii) recovering IL-1 million binding molecules from the culture. . The AAL160 antibody has been found to have the binding specificity of the human IL-ip epitope as the present invention, wherein the loop comprises mature human IL_i residues Gly 22, Pro 23, Tyr 24 and Glu 25. (Residues of mature human IL-1/? Giy 22, Pro 23, Tyr 24 and Glu 25 are each equivalent to residues of human IL-1 々 precursors 138, 139, 140 and 141). This decision appears to be outside the IL-1 receptor recognition site and, therefore, the most surprising is that antibodies at this epitope (such as the AAL160 antibody) inhibit IL-1々 binding to its receptor. Antibodies, in particular chimeric and CDR-grafted antibodies and in particular human antibodies, have the binding specificity of the mature human IL-1 Θ epitope, wherein the loop comprises Gly 22, Pro 23, Tyr 24 and Glu 25 residues, and can suppress jLq point -16 - This paper is ¥^ China National Standard (CNS) A4 specification (210 X 297 mm) (please read the note on the back and fill out this page) 1111111 IIIIIIII . 1314558 A7 B7 V. INSTRUCTIONS (14) The combination with its receptor; and the use of the antibody for the treatment of IL-mediated diseases and discomfort are novel methods and are included in the scope of the present invention. Thus, a further aspect of the invention comprises an IL_丨0 antibody having the antigen binding specificity of the human IL-1 cold epitope, wherein the loop comprises mature human IL-1/? Gly 22, Pro 23, Tyr Residues such as 24 and Glu 25 inhibit the binding of IL-1 /5 to its acceptor. In a still further aspect of the present invention, the method comprises the use of: 0 IL-1々 antibody, wherein the IL-1 Θ antibody has antigen binding specificity of the mature human IL-1々 epitope, wherein the loop comprises Giy 22, ΡΓ0 23. Residues such as Tyr 24 and Glu 25 which inhibit IL_1/? binding to its receptor for the treatment of IL-1/5 vector disease or discomfort; ii) a treatment for IL-1/? vector disease or discomfort A method of treating a patient comprising administering to the patient an effective amount of an IL-1 yS antibody having antigen-binding specificity of a mature human IL-1 epitope, wherein the loop comprises Giy 22, pr0 23, Tyr 24 and a residue such as Glu 25 and which inhibits the binding of IL_1/? to its acceptor; iii) a pharmaceutical composition comprising an IL-1 cold antibody having the antigen binding specificity of the mature human IL-1 epitope, wherein The ring comprises residues such as Gly 22, Pro 23, Tyr 24 and Ghi 25, and inhibits the binding of α-oxime to its acceptor, together with pharmaceutically acceptable excipients, diluents or carriers; Iv) the use of IL-1 antibody, which has a mature human ratio The antigen binding specificity, wherein the loop contains residues such as G1y 22, Tyr 24 and Glu 25, and can inhibit il-1 and its acceptor at -17- (please read the back of the back sheet and fill out this page). IIIIIII - —I! — !- Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperatives Printed 21 ( /V Grid 4 ) A s) N (c Pre-standard country in the country with moderate ruler paper PCT 97 1314558 Five Ministry of Economics wisdom Property Bureau Staff Consumer Cooperatives Printed A7 Description of the Invention (15 〇 used to prepare an agent that can treat IL_1/? vector disease or discomfort. In the description of the present invention, 'antibody' can inhibit IL-Μ binding&quot; The degree of inhibition of the combination with the corpuscle is substantially the same as that of the aaL16 〇 antibody, wherein the meaning of "the same degree of inhibition" is as described above. In the description of the present invention, the term "IL_! vector disease" includes all i, whether directly or indirectly. The disease and condition, including the cause, development, progression, persistence or pathology of the disease or condition. In the context of the present invention, the term "treatment" or "medical" refers to all prevention or treatment of disease and treatment. Or disease palliative treatment, including treatment of patients who are prone to or likely to contract such diseases, and those who have become ill or have been diagnosed with the disease or condition, and include inhibition of clinical recurrence. Mature human IL-1々 An antibody having an epitope binding specificity, wherein the 4-loop comprises residues Gly 22, Pro 23, Tyr 24 and Glu 25 and inhibits binding of IL-1 β to its corpuscle, hereinafter referred to as an antibody of the present invention. Preferably, the anti-system of the invention has an antibody that binds to the specificity of the human ILd0 in a natural (e.g., normal physiological condition) condition, rather than a denaturing condition (e.g., in the presence of a denaturing agent such as SDS). The antibody of the present invention can be reacted with non-human IL-1; the epitope contained in the non-human IL-1; 5s comprises Gly at residue 22, ρΓ0 at residue 23, Tyr and residue at residue μ Glu of μ, and it is very similar to the human decision bit. As the antibody of the present invention can cross-react with primate IL_i million s, such as rhesus monkey, macaque monkey IL_丨 or simian IL_i. Preferably, the anti-system of the invention is an anti-systematic human antibody of the invention, preferably an AAL160 antibody or a direct counterpart thereof, in accordance with the first and second aspects of the invention. -18 - This paper size is applicable to China National Standard (CNS) A4 specification (210 X 297 mm) f Please read the notes on the back and fill out this page) !314558 A7 B7 V. Inventions (16) (Read first Precautions for the back side. Fill in this page. The antibodies of the present invention block the action of IL-1々 on its target cells and thus display diseases and discomforts that can be used to treat IL-1 vectors. The pharmacological activities of these and other antibodies of the invention can be demonstrated by the following standard assays: 1 - Neutralizing neutralizing IL-1 /?-dependent cells of human IL-1/?-mediated IL-8 promoter gene activation The effectiveness of the signal is detected by communication genetic testing. The human melanoma cell line G361 is stably infected with the human IL-8 promoter group due to the transfer of the major luciferase communication gene structure. The communication gene expression and activity are based on IL-1 /5 or TNF 〇 of the cell line. In an adjusted medium, 300 μg/ml of recombinant human IL-1 or 1 〇〇 picogram/ml equivalent plus various concentrations of the antibody of the invention or IL-1 receptor antagonist (concentration between 6 And 18,000 pM) to stimulate the cells. The chimeric antibody, Simulect® (basiliximab), was used as a paired isomer control group. Insectic luciferase activity was quantified by chemiluminescence assay. In this assay, the antibodies of the invention typically have an IC5 〇 of about i nano equivalents (e.g., from about 0.2 to about 5 nano equivalents). 2. Primary human fibroblasts, pGe2 and interleukin _6 positive _ ^ 々 dependent production of neutralization, Ministry of Economic Affairs, Intellectual Property Bureau, employee consumption cooperative, printing of primary human ecdysis, tissue preparation, PCI* IL_6 It depends on IL-1々. The use of TNF-or alone does not effectively induce these inflammatory mediators, which must act synergistically with IL-1. The primary ecdysis fibroblasts are used as a surrogate for i _ induced cell viability. The adjusted medium obtained by recombinant IL-1/? or from human PBMCs stimulated by LPS plus various concentrations of the antibody of the present invention or IL1RA (concentration between 6 -19-

1314558, invention description (17) to 18, 〇〇〇 pM) stimulate the initial type 浐rn9 &lt; 4, secret cell tissue mother cells. The chimeric jade anti-CD25 antibody, sirrel, sputum, sputum, sputum, sputum, smear, smear, smear, smear, smear The antibody of the present invention typically inhibits IC5 produced at 1 L·1 point when stimulated for 6 hours or by RIA analysis of p(}E e ^ = taking the surface layer and analyzing it by brain A analysis. Approximately 1 nanometer to n ... nano equivalents, while inhibiting PGE production t Γ : lower (such as from about °, 1 to 2 production (IC5 〇 is about 1 nano equivalent (such as from, spoon 0.1 to about 1) Nano-equivalent). The antibody of the present invention can effectively block the cold action of IL-1. Therefore, the antibody of the present invention has the following pharmaceutical effects: The anti-system of the present invention can be used for prevention and treatment! Disease or disease = 'such as inflammatory conditions, allergic and allergic conditions, allergic reactions, autoimmune diseases, severe infections, and organ or tissue transplant rejection. The antibodies of the invention can be used to treat the heart, lungs, heart-lung, Recipients of liver, kidney, pancreas, skin or corneal transplantation, and prevention of graft-to-host diseases, such as conditions after bone marrow transplantation. The antibodies of the present invention are particularly useful for treating, preventing, or ameliorating autoimmune diseases and inflammation. The condition, specifically, the element of autoimmune Because of inflammatory conditions, such as arthritis (such as rheumatoid arthritis, chronic progressive arthritis and arthritis) and rheumatism 'including (four) inflammatory conditions of bone loss and wind /.&quot;, f disease, inflammatory pain , allergic reactions (including both respiratory allergies and skin allergies) and allergies. Specific autoimmune diseases in which the antibodies of the present invention can be used include autoimmune blood diseases (including, for example, hemolytic anemia, aplastic anemia, oligoerythrocyte anemia, and Primary thrombocytopenia), systemic lupus erythematosus, multiple chondritis, scleroderma, Wegener granuloma-20- This paper size applies to the Chinese National Standard (CNS) A4 specification (210 x 297 mm) (Please read the notes on the back and fill out this page)

Ministry of Economic Affairs, Intellectual Property Bureau, Staff Consumer Cooperatives, Printing 1314558

Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention description (18) disaster, dermatomyositis, chronic progressive hepatitis, myasthenia gravis, cognac, Stevenson-Johnson syndrome, primary Sexual tropical aphthous, autoimmune inflammatory bowel disease (including ulcerative colitis, Crohn's disease and allergic bowel syndrome), endocrine eye disease, Graves' disease, Sarcoidosis, multiple sclerosis, primary biliary cirrhosis, juvenile diabetes (type I diabetes, uveitis (pre and post), dry keratoconjunctivitis and spring keratoconjunctivitis, interstitial pulmonary fibrosis, cowhide Sputum arthritis and spheroid nephritis (with or without renal disease, including primary renal syndrome or minimal change nephropathy). The antibodies of the invention may also be used to treat, prevent or ameliorate asthma, bronchitis, pneumoconiosis Disease, emphysema, and other or respiratory inflammatory diseases. The antibodies of the present invention can be used to treat unpleasant causes than "media or with specific sputum production, especially Acute and hyperacute inflammatory responses associated with IL-1 cold, or TNF-promoted TNF release, such as acute infections such as septic shock (eg, endotoxic shock and adult dyspnea syndrome), meningitis, pneumonia; Severe burns; and treatment of cachexia or constipation associated with pathological TNF release, cancer, or dysfunction of the ear, especially AIDS-related cachexia, such as HIV infection or HIV infection Consequences The antibody of the present invention is particularly useful for the treatment of bone metabolic diseases, including osteoarticular spasm, osteoporosis and other inflammatory arthritis, and general bone loss, including age-related bone loss, and particularly periodontal disease. - This paper size applies to the S National Standard (CNS) A4 specification (210 X 297 mm) (please read the notes on the back and fill out this page) _装--------订·丨—丨1314558 A7 V. INSTRUCTIONS (19) Tumors The antibodies of the present invention can be used for the treatment of cancer, in particular, IL_丨-dependent swelling 0 (please read the notes on the back and then fill out this page). In these instructions, appropriate agents Obviously, it should be varied according to various conditions, such as the specific type of antibody of the present invention, the host, the mode of administration, and the nature and severity of the therapeutic condition. However, for prevention, it is daily from about 0.1 mg to 5 mg per kg of body weight. The dosage of the present invention is usually satisfactory. The antibody of the present invention can be conveniently administered parenterally, intravenously (e.g., before injection into the anterior elbow: vein or other peripheral vein), intramuscularly, or subcutaneously. Prophylactic treatment typically includes daily The molecules of the present invention are administered once to once a week for a total of 2 to weeks. The pharmaceutical composition of the present invention can be prepared by a conventional method. The composition of the present invention (4) is supplied in a cold ♦ dry form, and is dissolved in an appropriate form when it is used. Aqueous carriers such as sterile water for injection or sterile buffered saline. If you need to brew a large amount of solution for perfusion, such as intravenous injection, compared to a large number of shots, such as subcutaneous injection, preferably combined with human _, / serum albumin or the patient itself (added heparin blood into the physiological salt) Water is prepared. The presence of excess protein prevents the antibody from being adsorbed to the perfusion ^ 4.5 weight percent &lt;salt aqueous solution. Printed by the Ministry of Economic Affairs, Intellectual Property Office, Staff Consumer Cooperative, the following example steps are described for illustrative purposes only: Η 'F 尺 ΐ ΐ ΓΓ ΓΓ 1 1L_1 type 1 and type 11 acceptor pair AAU60 and IL_ 1 β &gt;, 'Sino-consistent inhibition; -22- Bu paper scale applies Chinese National Standard (CNS) A4 specifications (2) "Tear public love - 1314558 A7 B7 V. Invention description (20) Figure 2 shows Inhibition of IL-1々-induced fever by AAL160 in the rat model, and (please read the notes on the back and fill out this page) Figure 3 shows the effect of AAL160 on IL-1 induced fever in rats. The mouse of the transgenic gene produces antibodies against human IL-1, which are designed to express human IgG/λ: groups, but not the murine immunoglobulin group (Fishwild et al., 1996, Nat Biotechnol., 14, 845-851). Immortalized b cells from these mice were immortalized using standard fusion tumor technology to obtain murine fusion tumor cells that secrete human IgGI/κ antibody AAL160. Example 1: Production Fusion tumor and purified antibody economy The human employee consumption cooperative printed a recombinant human IL-1 (50 μg) placed in an adjuvant to immunize a number of genetically engineered mice 66 (Medarex Inc. Annadale, NJ). The mice were re-injected. Five times, and the last injection was performed 3 days before the fusion. On the day of fusion, the mice were sacrificed by inhalation of C〇2 and the PEG 4000 was used in an equivalent manner to ΡΑΙ-0 cells (the mouse myeloma cell line). Fusion of spleen cells (4.1X1 〇7). The fused cells were plated into 624 wells (1 ml/well), each containing a nutrient layer containing mouse peritoneal cells (Balb C mice) 'adding HAT RPMI 1640, 10% heating does not activate bovine fetal serum 5 X 10-5M /?-mercaptoethanol. The surface is collected and analyzed by ELISA and screened for IL-1 0 reaction monoclonal antibody. Five IgG/X times can be confirmed. Graded monoclonal antibody. Use 4 x 96-well micro-price plate for colonization, coating 〇5 -23 per tank. This paper scale is applicable to China National Standard (CNS) A4 specification (210 X 297 mm) 1314558 A7 B7 , invention instructions (21) cells. After two weeks, the cell was examined by reverse phase microscopy. The surface material was collected from the growing trough and the production status of the anti-IL-1 々 monoclonal antibody was evaluated by ELISA. 1-2 L of the adjusted surface material from the four sub-strains of the original confirmed fusion tumor #476 was prepared and utilized affinity. Chromatography purification of antibodies in protein A column. Purification of heavy and light chains and partial amino acid sequence amino acid sequencing by SDS-PAGE separation of purified anti-body AAL160 light chain and heavy chain and Eideman ( Edman) Degradation of the amine-terminated amino acid. The antibody used in these sequencing studies was 2 90% pure. The cDNA sequence encoding of the heavy and light chain variable regions is amplified by PCR and sequenced by PCR amplification of the mRNA of the selected fusion tumor cells. The heavy chain and light chain variable region amine terminus sequences and corresponding DNA sequences are detailed below, wherein the CDRs are shown in bold font. Sequence Identification Number 1 30 60

GAG GTG CAG CTG.GTG CAG TCT GGA GCA GAG GTG AAA AAG CCC GGG GAG TCT CTG AAG ATC

Glu Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Glu Ser Leu Lys lie 10 20 90 CDR1 120

TCC TGT AAG GGT TCT GGA TAC AGC TTT ACC AGC TAC TGQ ATC GGC TGG GTG CGC CAG ATG

Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Ser Tyr Trp lie Gly Trp Val Arg Gin Met Printed by the Intellectual Property Office of the Intellectual Property Office of the Ministry of Economic Affairs (please read the notes on the back and fill out this page) 30 40 150 CDR2 f 180

CCC GGG AAA GGC CTG GAG TGG ATG GGG ATC ATC TAT CCT AGT 6AC TCT GAT ACC AGA TAC

Pro Gly Lys Gly Leu Glu Trp Met Gly lie He Tyr Pro Ser Asp Ser Asp Thr Arg Tyr 50 60 210 240

AGC CCG TCC TTC CAA bGC CAG GTC ACC ATC TCA GCC GAC AAG TCC ATC AGC ACC GCC TAC

Ser Pro Ser P He G G G G G G G G G G G G G G G G G G G G G G G G G G G G G G Illustrative (22) Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed 270 300 CTG ChG TGG AGC AGC CTG AAG GCC TCG GAC ACC GCC ATG TAT TAC TGT GCG AGA TAT ACC Leu Gin Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys Ala Arg Tyr Thr 90 100 CDR3 330 AAC TGO GAT GCT TTT GAT ATC TGG GGC CAA GGG ACA ATG GTC ACC GTC TCT TCA Asn Trp Asp Ala Phe Asp lie Trp Gly Gin Gly Thr Met Val Thr Val Ser Ser Seq. Id No. 2 30 60 GAA ATT GTG TTG ACA CAG TCT CCA GCC ACC CTG TCT TTG TCT CCA GGG GAA AGA GCC ACC Glu lie Val Leu Thr Gin Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly Glu Arg Ala Thr 10 20 CDR1 90 120 CTC TCC TGC AGG GCC AGT CAG AGT QTT AGC AGC TAC TTA Leu GCC TGG TAC CAA CAG AAA CCT L Eu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Tyr Ala Trp Tyr Gin Gin Lys Pro 30 40 150 CDR2 180 GGC CAG GCT CCC AGG CTC CTC ATC TAT GAT GCA TCC KKC AGO GCC ACT GGC ATC CCA GCC Gly Gin Ala Pro Arg Leu Leu lie Tyr Asp Ala Ser Asn Arg Ala Thr Gly lie Pro Ala 50 60 210 240 AGG TTC AGT GGC AGT GGG TCT GGG ACA GAC TTC ACT CTC ACC ATC AGC AGC CTT GAG CCT Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Leu Glu Pro 70 80 270 CDR3 300 GAA GAT TTT GCA GTT TAT TAC TGT CAG CAQ CGT AGC Αλα TGO ATG TTC CCT TTT GGC CAG Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Arg Ser Asn Trp Met Phe Pro Phe Gly Gin 90 100 GGG ACC AAG CTG GAG ATC AAA Gly Thr Lys Leu Glu lie Lys -25- (Please read the notes on the back and fill out this page) .1 This paper size applies to the Chinese National Standard (CNS) A4 specification. (210 X 297 mm) 1314558 Intellectual Property Intelligence Bureau Staff Consumer Cooperative Printed A7 B 7 V. INSTRUCTIONS (23) The DNA sequence coding and corresponding amino acid sequence of the AAL160 heavy and light chain variable regions are also found in the accompanying Seq. Id nos. 1 to 4 sequences. Construction of heavy and light chain expression vectors The Vl and VH coding sequences are amplified by PCR and inserted into the cassette vector via appropriate restriction sites to provide immunoglobulin promoter genes, from RFT2 antibodies (Heinrich et al. 1989) J. Immunol. 143, 3589-97) Preamble sequence, J-fragment portion and junction supplier location. The light chain cassette comprising the entire VL region, the promoter gene and the leader sequence for secretion is transferred to an expression vector comprising a human Ck gene, an immunoglobulin heavy chain promoter and a modified murine dhfr cDNA for utilization. Methotrexate (MTX) selection. The heavy chain cassette is transferred to an expression vector which encodes a human IgG1 gene, an immunoglobulin heavy chain promoter and an anti-neomycin gene for selection. Both the heavy and light chains are located in an expression vector structure similar to the genomic structure of the re-arranged immunoglobulin genes, which are believed to be important for high-level gene expression. To produce antibodies, the vectors are co-transfected into a suitable host cell strain, such as the SP2/0 cell line, the cells containing the vector sequence are selected by methotrexate, and the selected cell line is cultured to express the AAL160 antibody. Alternatively, a GS-based amplification/selection system (as described in EP 0256055 B, EP 0323997 B or European Patent Application 89303964.4) may be used, in which case the dh code selectable marker should be replaced by the GS code sequence. Example 2: Biochemical and Biological Data-26- This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) (please read the notes on the back and fill out this page) - Pack---- ----Book --------- 1314558

The monoclonal antibody AAL 160 was found to neutralize the activity of interleukin] in vitro. The monoclonal antibody is further characterized by its Biacore analysis in combination with recombinant human IL-1 0. The mode of neutralization was assessed by a competitive binding study with Soluble-Receptor. The biological activity of the antibody AAL160 against recombinant and naturally produced IL-1 was determined in primary human cells (Example 3) for its response to IL-1々. ^ 2.1 Determination of Dissociation Equilibrium Constant The binding rate and dissociation rate constant of recombinant human IL_Wan and AAU6〇 were determined by the Bayesian analysis method. AAL16〇 was immobilized and the binding ratio of recombinant ratio-丨々 was measured by surface particle genomic resonance, which ranged from 〇5 to 12 nano equivalents. The selected version allows the association of IL-1 cold to AAL160 based on 丨:丨 stoichiometry. Use the BIA evaluation (BIAevaluation) software for the number of boards. (Please read the notes on the back and fill out this page.) Binding Rate Constant [NTV1] (n=15) (3.9110.14) xlO5 Average 値 + Standard Error Dissociation Rate Constant [s·1] (n=15) (1.53 +0.05) χ ΙΟ '4 average 値 + standard error dissociation balance often (n = 15) (396.6 ± 19.5) χ 1 〇 '12 average gentleman standard error -------------- Ministry of Economic Affairs The Intellectual Property Bureau employee consumption cooperative printed AAL160 and the recombinant human IL-i々 have a high affinity. 2.2. Competitive inhibition of soluble IL-1 receptor receptor binding. Competitive competition of soluble IL-1 type I and type 11 receptors. Determination of AAL160 and soluble human ratio-丨I type and work type by Bayeux method Accept the competition between the bodies. AAL160 was immobilized on the surface of the pellet and injected with recombinant human IL-1(R) (8 nano equivalents) in combination with AAL160 without addition or increasing concentration to add recombinant human soluble receptor (〇_1〇 nano equivalent) or acceptor ΙΙ (〇·80 nano equivalents). The results obtained are shown in Figure 1. Nvp 27- This paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm)

I Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 1314558 A7 _____B7__ V. Description of invention (25) The combination of AAL160 NX-1 and IL-1々 is competitive with IL-I receptor type I and type II. 2.3. Responses of rodents and monkeys to human il-1 alpha, human IL-1RA, and IL-1/3. AAL160 was determined by analysis of AAL160 against human IL-1 π, IL-1RA, and murine animals. , the reaction of rats, rabbits and macaques IL-1々. AAL160 was fixed and tested for cytokines at a concentration of 8 nano equivalents (if the tester was IL-1 (20 nano equivalents). Total binding rate + standard error human IL-1 100 human IL -lor 〇.7±0.7 (n=3) human IL-IRa 1.2±1.2 (n=3) mouse IL-1 2.8±1.5 (n=3) rat IL-1/5 3.0±2.5 (n= 3) Macaque IL-1; 5 96.4 ± 6.8 (n = 3) Rabbit IL-1 point 12.1 ± 2.3 (n = 4) AAL160 and human IL-la, human IL-IRa, or murine, rat or rabbit IL_1 The parental fork reaction is not obvious. The reaction of rhesus monkeys is the same as that of human cytokines. I,3: IL-1 ^-dependent production of PGE2 and interleukins in primary human fibroblasts -6 Neutralization The production of primary human ecdysplasia tissue cells (?) £2 and _6 is dependent on IL-1 million. TNF-π cannot effectively induce these inflammatory mediators alone, but requires -28

(Please read the precautions on the back and fill out this page) ▼ Install I---book-------^ 1314558 A7 V. Description of invention (26) Synergistic with IL-1. The primary ecdysis fibrous tissue mother cell line acts as a proxy model for IL_丨-inducing cell activation. (Please read the phonetic transcription on the back side and then fill out this page) Stimulate primary human fibroblasts with recombinant IL-1々 or from LPS-stimulated human pbmC-adjusted medium and add various concentrations of AAL160 or IL- 1RA (ranging from 6 to 18,000 pM). The chimeric anti-CD25 antibody, Westlaket (Baxilic), was used as a paired isomer control. Surfaces were taken 16 hours after stimulation and tested for IL-6 by ELISA or PGE2 with RIA. AAL160 1(:5〇±standard error (η.) IL-1 Ra IC50 soil standard error (n23) IL-6 secretion recombination 0.34±0,037 nano equivalents no IL-6 secretion concentrated medium 0.6±0.09 nano equivalent 0.03±0.001 nanoequivalent pge2 production concentrated medium 0.79±0·17 nano equivalents AAL160 was not detected to effectively prevent IL-6 and PGE2 from human ecdysone tissue cells, and the recombinant and native IL-ly5 IC5G were similar. 4: The effectiveness and duration of AAL160 in the body: The Ministry of Economic Affairs, the Intellectual Property Bureau, the employee consumption cooperative, printed the rat model, and injected intravenous IL-1 /5 (100 ng/mouse) with fever. While testing the anti-human IL-1 cold antibody, the in vivo efficacy of AAL 160. This antibody resulted in a dose-related inhibition of the febrile response at doses ranging from 1, 3 and 10 μg/kg intravenously (n=6). Mouse) - see Figure -29 - This paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) !314558

I Department of Economic Affairs Intellectual Property Bureau employee consumption cooperative printed. Using CHI 621 (Xi Mu Like, Xi antibody. ... Ksima) as a control group, investigating the role of IL_M_ induced fever of aal gas: pulse injection of humans!... Or 3 minutes (standard plan) and measure body temperature after 2 and 4 hours. A similar degree of inhibition of fever was observed in _ times (see Figure 3). For example, the control group antibody CHI621 (West Muckerett, Baxi Lixima) has no effect at two time points. The results show that the AAL16 〇 human antibody can be present in the active form for at least 24 hours in Dabai &amp; and will not metabolize during this period: in addition to or in combination in the tissue. The AAL160Fab structure was determined by a resolution of 2.0 and a resolution of AAL160Fab and its complex with IL_ly5. The growth of AAL160Fab was determined by 2.0 resolution. The growth was carried out from Fab crystals by vapor diffusion technique (pH 9.5, 50% PEG 200, 0.1 M CHES) Medium) 2. The best resolution data set (Rsym = 〇, 〇 51 'completeness = 99.9%, excess = 8.2). The crystal is a space group (space group^J^, unit lattice size is a=62.17A b= 89.83A c= 123.73 and a symmetrical unit contains one Fab molecule (Matthews coefficient: 3.6 A3/Da) , estimated solvent capacity: 66%). The structure was determined by the displacement method and refined to the final crystal R_factor of 0.209 (free R-factor = 〇.261). The final pattern contains residues 1-213 of the light chain, heavy Chains 1-131 and 138-218, 387 water molecules and 1 PEG molecule. In addition to the light chain Trp 94 (CDR3), the final electron density of all CDR residues is properly determined. Of the two crystal types, -30- This paper scale applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) ---------tr--------- rtt first Read the note on the back 4 and fill out this page.) 1314558 A7 ------— _B7______ V. INSTRUCTIONS (28) The position of the side chain of this residue is still undetermined. It is presumed that it is highly dynamic because it is not bound to the antigen. .

Crystallization of Fab Complex and Bi-Lu and the Preliminary Experimental Mode of the Complex: From 76 mg/ml i: 1AAL160 Fab and IL-1 Lu Antigen Complex Reserves (stored in 2.0 M ammonium sulfate, O .lM Tris pH 8.5) Crystallization of a few complexes. The crystal growth is very slow in a few weeks. The diffraction of its original source is weakly about 3.2 A. A preliminary set of data was collected and molecular replacements were attempted using the high resolution structure of the free Fab and human IL-1 (J. p. Priestie et al., EMBO J. 7, 339 (1988)) as the starting mode. When the Fv and Fc parts of Fab are used as separate modules, the calculation results can be obtained with a fairly clear and unambiguous analysis (after the AMORE FITTING step, the correlation rate is 67.1% 'R-factor 0.354, used The data is between 8.〇 and 〇3.5 A). Subsequent comparison of the free and combined forms of Fab revealed a very large difference in elbow angle between the two structures. The results of the molecular replacement calculations provide the first molecular pattern of interaction between the IL-1 antigen and the monoclonal antibody AAL160. A preliminary analysis of these interactions indicates that 1) IL-1 cold produces a close interaction with the CDRs of the three CDRs and the light chain of all heavy chains. In contrast, it exerts some interaction with the CDR2 of any of the CDR1 and light chains. 2) A loop comprising a residue such as Gly 22, Pro 23, Tyr 24 and Glu 25 of mature IL-1 is bound to the center of the antigen-binding site and thus becomes an important unit of the epitope. Interestingly, the ring is not located in the region where the molecule is the most different from IL-1 in mice. Pro 23, Tyr 24 and Glu 25 were retained, but residue 22 was Gly in human IL-1 and Asp in mouse IL-10. Comparing human (PDB into 2ilb) and mouse IL-1 y? (PDB entry 8ilb) crystal structure shows that the -31 - paper scale applies to China National Standard (CNS) A4 specification (210 X 297 mm) (please Read the precautions on the back and fill out this page. - Install---Order---------. Ministry of Economic Affairs, Intellectual Property Bureau, Staff and Consumer Cooperatives, Print I314558 A7 B7, Inventions (29) (Please Read the precautions on the back and fill out this page. The point mutations result in a very different main-chain pattern around the Pro 23. This local structural difference is consistent with the observed cross-reactivity between AAL160 deficiency and mouse cytokines. Sequence Listing &lt;11〇&gt; Novartis AG &lt;120&gt; Human IL-1 Antibody &lt;13〇&gt; 4-31289A &lt;160&gt; 4

&lt;170&gt; Patent (Patentln) version 3.0 &lt;21〇&gt; 1 &lt;211&gt; 354 &lt;212&gt; DNA &lt;213&gt; Mus musculus &lt;220&gt; 9. &lt;221&gt; CDS &lt;;222>(1)..(354)&lt;400&gt; 1 gag gtg cag ctg gtg cag tct gga gca gag gtg aaa aag ccc ggg gag 48

Glu Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 15 10 15 tct ctg aag ate tcc tgt aag ggt tct gga tac age ttt acc age tac 96 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative print

Ser Leu Lys lie Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Ser Tyr 20 25 30 tgg ate ggc tgg gtg ege cag atg ccc ggg aaa ggc ctg gag tgg atg 144

Trp lie Gly Trp Val Arg Gin Met Pro Qly Lys Gly Leu Glu Trp Met 35 40 45 ggg ate ate tat cct agt gac tct gat acc aga tac age ccg tec ttc 192

Gly lie lie Tyr Pro Ser Asp Ser Asp Thr Arg Tyr Ser Pro Ser Phe 50 55 60 caa ggc cag gtc acc ate tea gee gac aag tec ate age acc gee tac 240

Gin Gly Gin Val Thr lie Ser Ala Asp Lys Ser lie Ser Thr Ala Tyr 65 * 70 75 80 ctg cag tgg age age ctg aag gee teg gac acc gee atg tat tac tgt 288 -32- This paper scale applies to Chinese national standards (CNS ) A4 size (210 X 297 mm) 1314558 A7 B7 7; Description of invention (3〇)

Leu Gin Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 gcg aga tat acc aac tgg gat get ttt gat ate tgg ggc caa ggg aca 336 Ala Arg Tyr Thr Asn Trp Asp Ala Phe Asp lie Trp Gly Gin Gly Thr 100 105 110 atg gtc acc gtc tct tea 354 Met Val Thr Val Ser Ser 115 &lt;210&gt; 2 &lt;211&gt; 118 &lt;212&gt; PRT &lt;213&gt;Muscle muscle &lt;400&gt; 2 Glu Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 15 10 15 (Please read the note on the back and fill out this page)

Ser Leu Lys He Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Ser Tyr 20 25 30

Trp lie Gly Trp Val Arg Gin Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45

Gly lie lie Tyr Pro Ser Asp Ser Asp Thr Arg Tyr Ser Pro Ser Phe 50 55 60

Gin Gly Gin Val Thr lie Ser Ala Asp Lys Ser lie Ser Thr Ala Tyr 65 70 75 80

Leu Gin Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95

Ala Arg Tyr Thr Asn Trp Asp Ala Phe Asp He Trp Gly Gin Gly Thr 100 105 110

Met Val Thr Val Ser Ser 115 9. Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Print &lt;210&gt; 3 &lt;211&gt; 321 &lt;212&gt; DNA &lt;213&gt;Muscle &lt;220&gt;&lt;221&gt; CDS &lt;222&gt; (1)..(321) &lt;400&gt; 3 gaa att gtg ttg aca cag tct cca gcc acc ctg tct ttg tct cca ggg 48-33- This paper scale applies to China National Standard (CNS) A4 specification ( 210 x 297, mm) 1314558 A7 B7 V. Description of invention (31)

Glu lie Val Leu Thr Gin Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 15 10 15 gaa aga gcc acc etc tcc tgc agg gcc agt cag agt gtt age age tac 96 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Tyr 20 25 30 tta gcc tgg tac caa cag aaa cct ggc cag get ccc agg etc etc ate 144 Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu lie 35 40 45 tat gat gca tcc aac agg gcc act ggc Ate cca gcc agg ttc agt ggc ' 192 Tyr Asp Ala Ser Asn Arg Ala Thr Gly lie Pro Ala Arg Phe Ser Gly 50 55 60 agt ggg tet ggg aca gac ttc act etc acc ate age age ett gag cct 240 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr He Ser Ser Leu Glu Pro 65 70 , 75 80 gaa gat ttt gca gtt tat tac tgt cag cag cgt age aac tgg atg ttc 288 Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Arg Ser Asn Trp Met Phe 85 , 90 95 cct ttt ggc cag ggg acc aag ctg gag ate aaa 321 Pro Phe Gly Gin Gly Thr Lys Leu Glu lie Lys ' 100 105 &lt;210&gt; 4 &lt;211&gt; 107 &lt;212&gt; PRT &lt;213&gt; Muscle &lt;400&gt; 4 Glu lie Val Leu Thr Gin Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly X 5 10 15

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu lie 35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly lie Pro Ala Arg Phe Ser Gly 50 55 60 (Please read the note on the back and fill out this page) --I-----订-------1·- Ministry of Economic Affairs, Intellectual Property Bureau, employee consumption cooperative, printing

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Leu Glu Pro 65 70 · 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Arg Ser Asn Trp Met Phe 85 90 95

Pro Phe Gly Gin 'Gly Thr Lys Leu Glu lie Lys 100 105 -34 - This paper size applies to China National Standard (CNS) A4 specification. (210 x 297 mm) 1314558 A7 B7 Five Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative Description of the Invention (32) Sequence Listing &lt;1 10&gt; Novartis AG &lt;120&gt; Human IL-1 Antibody &lt;130&gt; 4-3 1289A &lt;160&gt; 4 &lt;170&gt; Patent Version 3.0 &lt;210&gt;&lt;211&gt; 354 &lt;212&gt; DNA &lt;213&gt; 目 肌&lt;220&gt;&lt;221&gt; CDS &lt;222&gt; (1).. (354) &lt;400&gt; 1 gag gtg cag ctg gtg cag tet Gga gca gag gtg aaa aag ccc ggg gag Glu Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Ly Gly Glu 15 10 15 ce 11 oa 12 gs ay a L 9 ute CL trce ccer gt cy cr taTV cr 9 e as crchoa T 3 tethtpcr 9 e as cr taTV ay 91 9G trce 5 ts 2 Jty 91 gG 9s aya L· tgg ate ggc tgg gtg ege cag atg ccc ggg aaa ggc ctg gag tgg atg Trp lie Gly Trp Val Arg Gin Met Pro Gly Lys Gly Leu Glu Trp Met 35 40 45 ggg ate ate tat cct agt gac tet gat acc aga tac age ccg tee ttc Gly Lie lie Tyr Pro Ser Asp Ser Asp Thr Arg Tyr Ser Pro Ser Phe 50 55 60 caa ggc cag gtc acc ate tea gee gac aag tee ate age acc gee tac Gin Gin Val Thr lie Ser Ala Asp Lys Ser lie Ser Thr Ala Tyr 65 70 75 80 ctg cag tgg age age ctg aag gee teg gac acc gee atg tat tac tgt Leu Gin Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95 geg aga tat acc aac tgg gat get ttt gat ate Tgg ggc caa ggg aca Ala Arg Tyr Thr Asn Trp Asp Ala Phe Asp lie Trp Gly Gin Gly Thr 100 105 110 atg gtc acc gtc tet tea Met Val Thr Val Ser Ser 115

&lt;210&gt; 2 &lt;211&gt; 118 &lt;212&gt; PRT &lt;400&gt; 2 Glu Val Gin Leu Val Gin Ser Gly Ala Glu Val Lys Lys Pro Gly Glu 1 5 10 15 Ser Leu Lys lie Ser Cys Lys Gly Ser Gly Tyr Ser Phe Thr Ser Tyr 20 ' 25 30 Trp He Gly Trp Val Arg Gin Met Pro Gly Lys Gly Leu Glu Trp Met -35· This paper size applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 6 9 (Please read the notes on the back and fill out this page.) 9. 1314558 A7 B7 V. Inventions (33) 35 40 45

Gly lie lie Tyr Pro Ser Asp Ser Asp Thr Arg Tyr Ser Pro Ser Phe 50 55 60

Gin Gly Gin Val Thr lie Ser Ala Asp Lys Ser lie Ser Thr Ala Tyr 65 70 75 80

Leu Gin Trp Ser Ser Leu Lys Ala Ser Asp Thr Ala Met Tyr Tyr Cys 85 90 95

Ala Arg Tyr Thr Asn Trp Asp Ala Phe Asp lie Trp Gly Gin Gly Thr 100 105 110

Met Val Thr Val Ser Ser 115 &lt;210&gt; 3 &lt;211&gt; 321 &lt;212&gt; DNA &lt;213&gt;Muscle muscle &lt;220&gt;&lt;221&gt; CDS &lt;222&gt; (1) . . (321) (Please read the notes on the back and fill out this page.) Ministry of Economic Affairs Intellectual Property Bureau employee cooperation cooperation Du-print clothing &lt;400&gt; 3 gaa att gtg ttg aca cag tct cca gcc acc ctg tct ttg tct cca ggg 48 Glu lie Val Leu Thr Gin Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 gaa aga gcc acc etc tcc tgc agg gcc agt cag agt gtt age age tac 96 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Tyr 20 25 30 tta gcc tgg tac caa cag aaa cct ggc cag get ccc agg etc etc ate 144 Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu lie 35 40 45 tat gat gca tcc aac agg gcc act ggc ate cca Gcc agg ttc agt ggc 192 Tyr Asp Ala Ser Asn Arg Ala Thr GXy lie Pro Ala Arg Phe Ser Gly 50 55 60 agt ggg tct ggg aca gac ttc act etc acc ate age age ett gag cct 240 Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Leu Glu Pro 65 70 75 80 gaa gat ttt gca gtt tat Tac tgt cag cag cgt age aac tgg atg ttc 288 Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Arg Ser Asn Trp Met Phe 85 90 95 cct ttt ggc cag ggg acc aag ctg gag ate aaa 321 Pro Phe Gly Gin Gly Thr Lys Leu Glu lie Lys 100 ♦ 105 &lt;210&gt; 4 i=211&gt; 107 &lt;212&gt; PRT &lt;213&gt;Muscle muscle v &lt;400&gt; 4 Glu He Val Leu Thr Gin Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 15 10 15 -36- This paper size is applicable to China National Standard (CNS) A4 specification (210 X 297 mm). Pack---1_!1 Order·!--. 1314558 a7 B7 V. Description of invention (34)

Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gin Ser Val Ser Ser Tyr 20 25 30

Leu Ala Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Arg Leu Leu lie 35 40 45

Tyr Asp Ala Ser Asn Arg Ala Thr Gly He Pro Ala Arg Phe Ser Gly 50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Leu Glu Pro 65 70 75 80

Glu Asp Phe Ala Val Tyr Tyr Cys Gin Gin Arg Ser Asn Trp Met Phe “ 85 90 . 95

Pro Phe Gly Gin Gly Thr Lys Leu Glu lie Lys 100 105 (Please read the note on the back and fill out this page)

I (K n n-TOJ _^i n n I Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 37- This paper scale applies China National Standard (CNS) A4 specification (210 X 297 mm)

Claims (1)

13 145 〇秘〇1〇〇267号 Chinese application patent scope i Patent application scope Φ本) A8 B8 C8 D8 91 9. Chromium; f yue yue 1. An IL-1/5 antibody with antigen binding specificity of mature human JL _ calling epitope, including ring (including Residues G丨y 2 2, Pro 23, Tyr 24 and Glu 25) and which inhibit il-1/3 binding to its receptor. 2. The IL_i cold antibody according to claim 1 of the patent application, comprising an antigen binding site comprising at least one immunoglobulin heavy chain variable region (Vh), the variable region comprising, for example, a sequence identification number: 1 The supervariable region CDR1, CDR2 and CDR3', that is, the CDR1 has the amino acid sequence Ser-Tyr-Trp-Ile-Gly, and the CDR2 has the amino acid sequence ile-ile_ Tyr-Pro-Ser-Asp-Ser- Asp-Thr-Arg-Tyr-Ser-Pro-Ser-Phe-Gln-Gly' and the CDR3 has an amino acid sequence of Tyr-Thr-Asn-Trp-Asp-Ala-Phe-Asp-Ile 0 3 ·According to The iL-lyS antibody of claim 1, which comprises an antigen binding site comprising at least one immunoglobulin heavy chain variable region (VH), the variable region CDR1, CDR2 and CDR3, wherein The variation region 〇0 ft1, €0112, and 〇113 overall is at least 80% homologous to the corresponding hypervariable region as shown in SEQ ID NO: 1:; and (ii) it inhibits IL-1 calling and its acceptor Binding, the inhibition procedure is the same as that of the IL-1 /3 antibody described in (i), and the supervariant regions CDH1, CDR2 and CDR3 (which are identical to the sequence identification number: 1 a reference molecule of the corresponding hypervariable region. 4. The IL-1 0 antibody according to claim 1 of the patent application includes both a heavy chain variable region (VH) and a light bond variable region (VL), wherein The IL-1 antibody includes 68528-970916.doc This paper scale applies to the Chinese National Standard (CNS) A4 specification (210X 297 mm) 1314558 a. Patent application scope At least one antigen binding site comprising: (i) an immunoglobulin heavy chain variable region (VH), the variable region of which comprises the mutated region CDR1, CDR2 and CDR3 as shown in SEQ ID NO: 1 That is, the CDR1 has the amino acid sequence Ser-Tyr-Trp-Ile-Gly, which has the amino acid sequence Ile-Ile-Tyr-Pro-Ser-Asp-Ser-Asp-Thr-Arg-Tyr-Ser- Pro-Ser-Phe-Gln-Gly, and the CDR3 has an amino acid sequence Tyr-Thr-Asn-Trp-Asp-Ala-Phe-Asp-Ile; and (ii) an immunoglobulin light chain variable region ( VL), which comprises CDR3 as shown in SEQ ID NO: 2, a supervariant region, i.e., CDR3 1 having an amino acid sequence 歹丨J Gln-Gln-Arg-Ser-Asn-Trp-Met_Phe-Pro. 5. The IL-1 cold antibody according to claim 1 of the patent application, comprising both a heavy chain variable region (VH) and a light chain variable region (V1), wherein the IL-1 cold antibody comprises at least one antigen binding site, These include: (i) an immunoglobulin heavy chain variable region (VH), the variable region of which includes the mutated region CDR1, CDR2 and CDR3, and (Π) - immunoglobulin light chain variable region (VL) , which comprises a CDR31 hypervariable region, (a) wherein the hypervariable regions cDR1, CDR2, CDR3 and CDR3 are at least 80% homologous to the corresponding hypervariable regions as shown in SEQ ID NO: 1 and 2; b) it inhibits the binding of IL - /3 to its receptor, and the degree of inhibition is the same as that of the IL-10 antibody described in (a). 68528-970916.doc _2 - The paper scale is suitable for ® Standard (CNS) A4 specification (21GX 297 mm). '------ 1314558 - ___ C8 ----- D8_ VI. Application for filial piety' domain and constant part and hypervariable region CDRl, CDR2, CDR 3 and the reference molecule of CDR 3 ' (which is identical to the corresponding hypervariable region indicated by the sequence identification number: i). 6. The condition of claim 4, "the stone antibody, wherein the immunoglobulin fe chain variable region (VL) comprises the mutated regions CDR1', CDR2, and CDR3 as shown in SEQ ID NO: 2, , the CDR1, having an amino acid sequence Arg-Ala-Ser-Gln-Ser-Val-Ser-Ser-Tyr-Leu-Ala', the CDR2 having an amino acid sequence Asp_Ala_Ser_Asn-Arg-Ala-Thr, and CDR3i has the amino acid sequence Gln-GLn-Arg-Ser-Asn-Trp-Met-Phe-Pro. 7. The IL_丨 cold antibody according to claim 4, wherein the immunoglobulin light chain variable region ( VL) includes the mutated region CDR1, CDR2, and CDR3', respectively, (1) wherein the supervariant region CDR1, CDR2, CDR3, CDR1, CDR2, and CDR 3' are at least 80% homologous to sequence recognition. Corresponding to the hypervariable region shown in Nos. 1 and 2; and (ii) inhibiting the binding of IL-1 cold to its acceptor, as if it had the same framework region as the IL_1/5 antibody described in (i) Constant portion and hypervariable region CDR1, CDR2, CDR3, CDR1', CDR2' and CDR3' (which are identical to the corresponding hypervariable region shown in Sequence Identification Numbers: 1 and 2) Reference molecule 8. A cold antibody as claimed in claim 1 which is a human antibody. 9 · According to the scope of the patent application, item ϊ L _ 丨 antibody, which includes at least one antigen binding position, 68528- 970916.doc This paper scale applies to the Chinese National Standard (CNS) A4 specification (210X297 public Chu:) AB c D 1314558 VI. The scope of the patent application (which includes a display with the sequence identification number 1 starting at position i and finally Position 1 1 8 amino acid sequence of the same amino acid sequence of the first region, or (11) which includes (a) heavy chain 'which includes and has been shown in the sequence identification: 1 starting from the amino acid a variable region of the amino acid sequence at position 1 and finally having the amino acid sequence of the amino acid position 1 1 8 and a strange portion of the human heavy chain; and (Μ light chain, which includes and displays the sequence identification No.: 2 The variable region of the amino acid sequence starting from the amino acid position 1 and finally having the amino acid sequence of the amino acid position of 10 7 and the constant portion of the human light chain. 10_ A preventive or therapeutic treatment The IL_i point is involved in the action of the disease or condition of the medical group a' Included is an IL _ 1 ^ antibody according to claim 1 of the patent application, and a pharmaceutically acceptable excipient, diluent or carrier. 11. A first DNA construct encoding a heavy chain or a fragment thereof, comprising a a first portion encoding a variable region comprising a selective backbone and a hypervariable region, wherein the sequence of the hypervariable region is cdr1, CDR2 and CDR3, and the amino acid sequence is shown in sequence identification number 1; a portion begins with a codon encoding the first amino acid of the variable region and finally a codon encoding the variable region final amino acid, and b) - encoding the heavy chain constant region or a fragment thereof The second part begins with a codon encoding the first amino acid of the heavy chain constant region and finally a codon encoding the final amino acid of the quirks region or a fragment thereof, and finally a stop codon. 68528-970916.doc This paper scale applies to the Chinese National Standard (CNS) A4 specification (210 X 297 mm) 1314558 A8 B8 C8 D8 Patent Application 12. A second dna structure encoding a light chain or a fragment thereof, Including a) - a first portion of the variable region encoding the variable region, the region comprising a selective backbone and a hypervariable region; the hypervariable region is (: 1) 1^3| = two: to be CDR1, and CDR2i 'The amino acid sequence is shown in SEQ ID NO: 2; the first part begins with a codon encoding the variable region first amino acid and finally encodes the codon of the variable region final amino acid, and Soil b) - a second portion encoding a constant region of a light chain or a fragment thereof, which begins with a codon encoding a first amino acid of the light chain constant region and which is capable of encoding a constant region or a fragment thereof, a final amine group The acid codon is finally relinked to the stop codon. 13. An expression vector which can be replicated in a prokaryotic or eukaryotic cell line, which comprises, for example, a d N A construct according to claim 11 or 12. ', 14. A method of preparing an IL-1 /3 binding molecule, comprising: cultivating an organism comprising at least one DNA construct according to claim 1 or 12 of the patent application and Prokaryotic or eukaryotic. Transgenic expression of vector replication; and (ii) Recovery of IL_i$ binding molecule from culture 0 68528-970916.doc This paper scale applies to Chinese National Standard (CNS) A4 specification (210X297 mm)