TW201217528A - Receptor binding agents - Google Patents

Abstract

Featured herein are non-naturally occuring cytokine domains that can be used, inter alia, to modulate cellular signalling responsive to interleukin-1 receptor I (IL-1RI), to treat disorders, and to detect and/or bind to cellular receptors, as well as other agents. Exemplary cytokine domains can contain amino acid residues from at least two parental cytokines domains, for example, receptor binding features, surface features, β strands, and loops from at least two parental cytokines domains.

Description

201217528 VI. Description of the invention: [Delivery of the stipulations of the stipulations of the stipulations of the stipulations of the stipulations of the stipulations of the stipulations of the stipulations of the stipulations of the stipulations of the stipulations of the stipulations U.S. Provisional Patent Application Serial No. 61/436,184, filed on Jan. 25, 2011, and U.S. Provisional Patent Application No. 61/ filed on Jun. 6, 2011. Priority is claimed to 493, 967, the contents of which are incorporated herein by reference. FIELD OF THE INVENTION This invention relates to a receptor binding agent.

L·J BACKGROUND OF THE INVENTION Interleukin-Ια (hereinafter IL-la) and interleukin-1β (hereinafter IL-Ιβ) are prototypic members of the immunoregulatory interleukin family (pr〇t〇typic members), and There are several prominent roles in regulating the immune system. IL-la binds to IL-Ιβ to interleukin-1 receptor 1 (hereinafter IL-1RI), resulting in a second receptor, the interleukin-1 receptor accessory protein (hereinafter Engagement of IL-lRAcP). The signal agonized by IL-la and IL-Ιβ leads to amplified T cell responses, including proliferation and survival of naive T cells and development of TH17 cells. C SUMMARY OF THE INVENTION 3 SUMMARY OF THE INVENTION 201217528 The present invention is characterized by a non-naturally occurring interleukin region, which can be used, inter alia, to be regulated by an interleukin-receptor (IL- 1RI)? Picking up cell signals, treating diseases, detecting cell receptors and/or binding to cell receptors, and other agents. In one aspect, the invention is characterized by an isolated protein comprising an interleukin region comprising amino acid residues from at least two parenteral regions, for example, from at least two Receptor binding characteristics, surface features, beta strands and loops (i00pS). In some embodiments, the interleukin region binds to IL_1RI and includes receptor binding characteristics from different parenteral regions, for example, from a receptor agonist to a receptor antagonist (such as 11^ 10 and ratio _11^, or IL-la and IL-IRa) 'from IL-Ιβ and IL-la, or from iL-lRa, IL-la and IL-IRa all. These receptor binding characteristics may correspond to residues, segmems, regions located at sites A and B (Sites A and B). Regarding these in the context of IL-UIL-Ιβ, IL-la, and iL_1Ra), the residues, segments and zones corresponding to the sites A and B are as defined below. With respect to site A (Site A), the interleukin region may have; (1) a site A residue which is at least 60, 70, 80, 85, 88, 90, 92, 95, 98, 戍 100 °/. Same as a corresponding residue on a first parental interleukin region; (ii) an extended site A residue (Extended Site A), which is at least 60, 70, 80, 85, 88, 90 '92, 95 ' 98 , or 100% identical to the corresponding residue on a -1 - parental region; (iii) Part A1 and A2, at least 8, 85, 88, 90, 92, 95 or 100% identical In the first intercellular region, the pair of 201217528 zone; and the (iv)-site a zone, which has at least 8〇, 85, 88, 9〇, 92 '95 or 10G% identical to the - Corresponding zone of the parental region. Regarding Site B (Site B) 'The interleukin region may have: (1) a site B residue that is at least 60, 70, 80, 85, 88, 90, 92, %, %, or 100% identical to - second Corresponding residues on the parenteral region; (9) extended site B residues, at least 60, 70, 80, 85, 88, 9 (), 92, 95, 98 ' or 100% identical to a second female Corresponding residues on the interleukin region; (10) M at position B, B2 and B3, at least 80, 85, 88, 9 〇, %, 95 or 1 〇〇/β identical to a second cytokine The corresponding zone of the zone; and (iV) a zone B zone of at least 8 〇, 85, 88, 90, 92, 95 or 100 〇/〇 identical to the corresponding zone of the second parent cell. In some embodiments, the interleukin region has the above characteristics (a)(1) and b(〇, (a)(ii) and b(ii), (4)(iii) and (b)(iii) or (4)(iv And (b) (iv), for example, wherein each feature further defines 80, 85, 88, 90, 92, 95, or 100% identity. For example, the first parental interleukin region can be IL_1 (3, And the second parental interleukin region can be IL-IRa. For example, the first parental interleukin region can be IL_la, and the second parent interleukin region can be IU1Ra. X, phytol The region may also include an amino acid from a second parental interleukin region located at one or more locations in the region, which attenuates interaction with a second receptor (eg, IL-lRAcP). Whereas, the '5-Second-Seminary Intercellular Interleukin region is IL-1 Ra. In some embodiments, the Xuan and Tian interferon regions include one or more regions from the IL-IRa, C and/or For example: C1 'to, D2, D3, D4, and/or D5), or at least 80, 201217528 85, 88, 90, 92, 95, or 100% identical to the sequence of the segments. For example, the cell Interleukin region includes (1) one a residue at position C which is at least 60,70 '8〇' 85,88,90,92,95,98, or 100% identical to the corresponding residue on IL-IRa; (Π) a residue at position D, at least 60,70,80,85,88, 90 '92 '95,98, or 100% identical to the corresponding residue on IL-IRa; (out) a Cl segment, at least 70,75,80,85,88 , 90, 92, 95, 98, or 100% identical to the corresponding residue on IL-IRa; or (iv) - D2, which has at least 3 '4 or 5 residues and a corresponding residue on IL-IRa The same is true. The interleukin region can include features (1) and (Π), or (ii) and (iii), for example, wherein each feature further defines 80, 85 '88, 90, 92, 95, or 100% The degree of identity ' or (iii) and (iv) the region may comprise a zone from at least two different human jLq family of interatrial regions, wherein the zone is selected from the zone A (with ^ Section A2) 'The B zone (with B1, 82 and 63 segments), the c zone, and the group of zones (having m, D2 'D3, m, and 〇5 segments). The interleukin region may comprise a region A from a different interleukin region Part B zone. The human zone of this site may be derived from nature: a steroidal agonist or antagonist; the sputum zone may be derived from a natural scorpion receptor agonist. The site of the naturally occurring receptor, the C-band, and the domain, is derived from the naturally occurring receptor site D1?»憨. a part of the stem J, which may be a chimeric region having at least 5'6'1()'15'2(), or 25 amino acid lengths and 85 '88 '90,92 ' 95, or 100% identical to the corresponding segment from at least two 6 201217528 interleukin regions, such as the first and second parental interleukin regions. The parental interleukin region may be an IL_1RI binding interleukin, such as IL-Ιβ, IL-la, and lL_1Ra. In some embodiments, the amino acid from the segment of the first parent cell is not derived from two or more other parenteral regions. In some embodiments, the interleukin region comprises at least two segments having a length of at least 5, 6, 10, 15, 20, or 25 amino acids, such as at least 8〇, 85, 88, 90' 92 '95, or 1% is the same as the corresponding segment of the first parental interleukin region, and includes amino acids not present on the segments, which are significant (eg, at least 50 '60, 70, 80, 85, 88, 90, 92, 95, or 100%) is identical to the corresponding residue on the second parental interleukin region. In some embodiments, the interleukin region comprises (i) at least two segments having a length of at least 5, 6, 10, 15, 20, or 25 amino acids, such as at least 80 '85, 88, 90, 92 '95 ' or 1〇0% is identical to a corresponding segment of a first parenteral region, and (ii) at least one, two or three segments, for example, having at least 5 ' 6 ' 7 ' 8 ' 10 , or 15 amino acid lengths, which are identical to a second parental interleukin region. For example, the interleukin region can include a first segment having 20 to 50 '25 to 50' 30 to 45, or 30 to 40 amino acids (eg, 29, 30, 31 ' 32 ' 33 ' 34 ' 35 ' 36 ' 37 ' 38,39 ' or 40) 'with a second segment' which has 20 to 45, 20 to 40, 25 to 40, or 25 to 35 amino acids (eg 25 ,26,27 '28' 29,30,3 Bu 32 '33,34, or 35), each identical (or at least 80,85,88 '90,92,95, or 98% identical) A parental interleukin region (eg, IL-1RA), identical to a third segment, 201217528 (or at least 80, 85, 88, 90 '92, 95' or 98% identical) a second female Interleukin region (eg IL-Ιβ or IL-la). For example, the third segment can be between 55 and 90, 60 to 90, 60 to 85' or 70 to 85 amino acids, for example: 75, 76, 77, 78, 79, 80 '81 ' 82 ' 83 ' 84, or 85 amino acids in length. In some embodiments: the first segment can be at least 80 '85' 88, 90, 92, 95, 98, or 1〇0% identical to WDVNQKTFYLRNNQLVAGYLQGPNV (SEQ ID NO: 9), here Also referred to as the A1 segment, and corresponds to a segment of the 16th to 40th residues of the sequence recognition 唬 No. 3 (SEQ ID NO: 3) and the 11th to 36th residues according to the 1L-1·3 number. The second segment can be at least 80 '85 '88, 90 '92, 95, 98, or 100% identical to sequence identification number 3 (SEQ ID NO: 3) (IL-IRa) 120-140 or The 120-141 residue corresponds to one of the 121 to 139 or 121 to 140 residues (according to the IL-Ιβ number). The third segment may be at least 80, 85, 88, 90, 92, 95, 98' or 100% identical to iL-Ιβ (SEQ ID No. 1 (SEQ ID ΝΟ: 1)) 45th to 100% or 42~120 residues. In some cases, the first segment may be at least 80, 85, 88 '90, 92, 95, 98, or 100% identical to the 14th to 45th of the sequence identification number 3 (SEQ Mountain NO: 3) A segment of the base (corresponding to residues 9 to 41 according to the IL-Ιβ number). The second stretch may be at least 8 〇, 85 '88, 90, 92, 95 ' 98, or 100% identical to the 12th to 145th residues of SEQ ID NO: 3 (corresponding to To one of the 12th to M7 residues of the SEQ ID NO: 3 (corresponding to the residues 121 to 147 according to the up number) according to the number 121 to 145 residues) or the sequence identification number 3 (SEQ ID NO: 3). In some embodiments, at least

8 201217528 1 Bu 41 and 120~147 residues (according to IL-Ιβ numbering) are collectively at least 80, 85, 88, 90 '92, 95, 98, or 100% identical to IL-IRa Residues. In some embodiments, the terminal of one of the segments of the first IL-1 family interleukin region is located at the 41st amino acid of SEQ ID NO: 1 (SEQ ID NO: 1) Within the fifth, fourth, third, or one amino acid, and the terminal line of one of the segments of the first IL-1 family of interleukin regions is located in SEQ ID NO: 1 (SEQ ID N0: 1) The fifth, fourth, third, second or one amino acid of the 121th amino acid. In some embodiments, the region comprises a segment having an N-terminus, a fifth, four, three, two or one of the 42th amino acid of Sequence ID #1 (SEQ ID NO: 1) Within the amino acid, and a C-terminus, within the fifth, fourth, third, second or an amino acid of the 120th amino acid of SEQ ID NO: 1 (SEQ ID NO: 1), And/or a segment having an N-terminus within the fifth, fourth, third, second or an amino acid of the 121th amino acid of Sequence ID No. 1 (SEQ ID NO: 1), and The C-terminus is located within the fifth, fourth, third, or one amino acid of the 145th amino acid of SEQ ID NO: 1 (SEQ ID NO: 1). In some embodiments, the residues in the region corresponding to positions 11 to 41 and 120 to 147 (according to the IL-Ιβ number) are collectively at least 80, 85, 88, 90 ' 92 ' 95, 98, or 100. % is identical to the corresponding residue on IL-IRa. This region may also be based on sequences from members of the IL-1 interleukin family, for example: located at a similar position as described above. In some embodiments, the interleukin region comprises one, two, three or more of the following 9 201217528 sequences: WDVNQKTFYLRNNQLVAGYLQGPNV (Sequence 歹 ij No. 9 (SEQ ID NO: 9)); NLEEK (sequence) Identification No. 10 (SEQ ID NO: 10)); RIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEK (SEQ ID NO: 11 (SEQ ID NO: 11)); AMEADQP (SEQ ID NO: 12); FLCTAMEADQPVSLTNMPDEGVMVTKFY (SEQ ID NO: 13); and/or at least 80, 85, 88, 90, 92, 95, 98, or 100% identical to the sequence of the above sequence. In some embodiments, the interleukin region comprises one, two, three or more of the following sequences: VQGEESNDKI (Sequence 歹 ij No. 14 (SEQ ID NO: 14)); KKKMEKRF (Sequence 歹 4 Identification No. 15 (SEQ ID NO: 15)); and FSMSFVQGEESNDKIPVALGLKEKNLYLSCVLKDDKPTL QLESVDPKN YPKKKMEKRFVFNKIEINNKLEFES (SEQ ID NO: 16); and/or at least 80, 85, 88, 90, 92, 95, 98, or 100% identical to the sequence of the above sequence. In some embodiments, one or more or all of the β1β2, β2β3, β8β9, and βΙΟβίΐ loops are at least 80, 85, 88, 90, 92, 95, or 100% identical to an IL-1 antagonist. For example: il-1 Ra, which corresponds to the ring. In some embodiments, one or more or all of the β4β5, β5β6, β6β7, and β7β8 loops are at least 80, 85, 88, 90, 92, 95, or 100% identical to a different IL-1. The pair of loops of the family interleukin region, but not the human interleukin that is closest to the β1β2, β2β3, β8β9 and βι〇βη loops. For example, § 'β4β5 'β5β6 'β6β7 and β7β8 loop s 10 201217528 one or more or all of at least 80, 85, 88, 90, 92, 95, or 100% identical to IL-1 efficacies These rings of agents (eg IL-Ιβ). In some embodiments, the β11β12 loop is at least 80, 85, 88, 90, 92, 95, or 100% identical to the corresponding loop from an IL-1 antagonist (e.g., IL-1Ra). In some embodiments, the interleukin region comprises at least 80, 85, 88 '90 '92 '95, or 100% identical to one of IL-lRa beta β2, β3, βΐ〇 and βΐΐ, two , a sequence of three or all. In some embodiments, the interleukin region comprises at least 80, 85, 88, 90, 92, 95, or 100% identical to one of β4, β6, β7, and β8 of IL-Ιβ, both, three Or all, or the sequence of β4, β5, β6, β7 and β8 of IL-Ιβ. In some embodiments, the interleukin region does not comprise greater than 80, 85, 90, 95, or 100% amino acid 146 to G59 'A55 identical to SEQ ID NO: 3 (SEQ ID Ν 0: 3) -G59, Α55~V83, 160~V83, Ν84~D95, 146~S110, V49-S110, or 146~G118 one segment. In some embodiments, the interleukin region does not comprise greater than 80, 85, 90, 95, or 100% amino acid hydrazone 7 to V41, R11 identical to SEQ ID NO: 1 (SEQ ID Ν 0: 1) ~Μ36, Ν102~D145, or Υ121~D145 one segment. In general, this interleukin region does not exist naturally. It differs from the human IL-1 family of interleukin regions. For example, it is less than 98, 95, 90, 85, 80, 75, 70, 65, 60, or 55% identical to IL-1Ra (SEQ ID NO: 3), IL -lp (SEQ ID NO: 1 (SEQ ID NO: 1)), and/or IL-la (SEQ ID NO: 2 (SEQ ID NO: 2)). The interleukin region may also be at least '40', 45, 50'55'60'65' For example, the chimeric region may be identical to IL-IRa 'IL-Ιβ' and IL-1 α between 30 and 95%, 40 to 90%, or 45 to 85%. For example, the transfer region may be the same as IL-Ιβ between 40 and 90% and the same as IL-IRa between 35 and 85%; the same as IL-Ιβ and between 45 and 80%. ~80°/〇 is the same as IL-IRa; between 45 and 72% is the same as IL-Ιβ and between 45 and 80% is the same as IL-IRa; between 45 and 72% is the same as IL- Ιβ and between 53 and 80% are identical to IL-IRa; between 50 and 72% are identical to IL-Ιβ and between 53 and 70% are identical to IL-IRa; between 60 and 72% are identical to IL-Ιβ is the same as IL-IRa between 53 and 68%; is identical to IL-Ιβ between 65 and 72% and is identical to IL-IRa between 54 and 60%; or between 68 and 72% It is identical to IL-Ιβ and is the same as IL-IRa between 54 and 57%. For example, the chimeric region may be the same as IL-la between 40-90% and the same as IL-IRa between 35-85%; at 40-80°/. Between the same as IL-la and between 45 and 80% identical to IL-IRa; between 45 and 72% identical to IL-la and between 45 and 80% identical to IL-IRa; at 45~ 72% is identical to IL-la and between 53 and 80% is identical to IL-IRa; between 50 and 72% is the same as IL-la and between 53 and 70% is identical to IL-IRa; 60~72% is the same as IL-la and between 53-68% is the same as IL-IRa; between 65~72% is the same as IL-la and between 54~60% is the same as IL-IRa Or between 68 and 72% identical to IL-la and between 54 and 57 〇/〇 identical to iL-lRa. The inhibitor comprises an IL-1 family interleukin region different from IL-IRa and will include a site C and/or site D having a characteristic of a naturally occurring receptor antagonist (eg, IL-IRa). Bind to the receptor. For example, the degree of identity between the region and human IL-Ιβ and /IL-1Ra is less than 98, 95, 92, 90, 12 201217528 85, and 80%. For example, the region is between 4〇~95%, 4〇~9〇% or 45~85% identical to IL-IRa. The region may also be between 4 〇 and 95%, 4 〇 to 9 〇 % or 45 to 85% identical to the IL-丨 family of interleukin agonists, such as chemido- or IL-Ιβ. In some embodiments, the interleukin comprises: at least 4 〇, 45, 50 ' 55 ' 60 ' 65 ' 70, 75, 80 or 85% of the amino acid from a cytokine signaling effect One of the first parental interleukin regions. In some embodiments, the interleukin region has the same greater amino acid than a receptor antagonist (IL-1 Ra) for a receptor agonist (eg, IL_丨P or IL_la) Degree (eg, at least 5, 10, 15 or 2%), but the interleukin region acts as an antagonist of IL_1RI. In some embodiments, the interleukin region is fully chimeric, for example, each of the amino acid groups in the region is from one of the parental interleukin regions, eg, from two parental cells. One of the prime regions or one of two or more parental interleukin regions. For example, the parental interleukin region is a human interleukin region or a non-human primate cytokine region. In some embodiments, the intercellular region is partially chimeric, e.g., not all of the amino acids in the region are from one of the parental interferon regions. For example, the protein binds to IL-1RI and modulates the signaling produced by the receptor', e.g., agonizing or antagonizing the signaling activity of the IL-1RI receptor. In some embodiments, when the protein contacts an IL-1p responsive human cell, the protein does not substantially induce IL-6 production, and/or the protein does not substantially induce IL-Ιβ reactivity reporter Gene production, for example, at 10 μg/ml hg/ml), 1 μg/ml 13 201217528 hg/ml, or 1 mg/ml (mg/mi). In general, the protein can be lower than the IC50 of just [...^" nanomolar concentration (4), inhibiting the signal transmission by IL-Ιβ (eg, in nanograms per milliliter (ng/mi)) The concentration, as described in the cytology assay below. The protein may be lower than the inhibition for IL-IRa, eg, at least Η), 20, or 50% lower. To the extent that signal transmission by il__ is inhibited. In certain embodiments, the interleukin region binds, for example, to the same or better affinity than one of the parental interferon regions. In some embodiments, the inhibitor has a concentration of less than 100, 5 Å, 2 Å, 1 〇, 5 ' or 1 nanomolar (nM) or less than 500 '400, 1 〇〇, or 5 〇 Pimo The KD value of the ear concentration (pM) is bound to IL-1RI. For example, the binding constant may be greater than Ιχίο4 ' 3χ 104, lxl 〇 5 ' or lxi 〇 6M-ls-1, and the dissociation may be lower than lxlO-3, Lxl〇·4,6χ10·4, or 6xl〇_5S-1. In some embodiments, the inhibitor has better affinity than Zha-ΐβ or IL-IRa (example) : lower KD) and/or binding to IL-1RI at a slower dissociation rate. For example, the interleukin region can bind to IL-1RI at a rate lower than or equal to the dissociation rate of IL-IRa. And/or bound to il-1RI with a binding constant greater than or specific for binding to IL-1 β. The interleukin region may be approximately between 12〇~180, 140~170, or 148~160' or 150 to 156 amino acid lengths. In some embodiments, the domain is 152 or 153 amino acid lengths. Typically the region comprises at least 10' 11 ' or 12 beta strands and is stably folded In some embodiments, the § interleukin region has a minimum of 38, 40, 42, 44, 46, 48, 50, 14 201217528 52 ' 54 ' 56 ' 58 ' 60 ' 62, or 64 ° C Tm value, as described below. It may have a boundary of 51-6 b 51~66, 56~6 b or 56~66. (: Tm value. In some embodiments 'the interleukin region until at least 48, 50, 51, 55, 57, 58, or 59C will begin to unfold (unf〇iding). For example, it has a Tm value 'The Tm value is at least one physiological buffer The Tm value of IL_1Ra and/or IL-Ιβ is within 1 (TC or 5 ° C. In some embodiments, it is more hot than IL-1Ra and/or iL-ΐβ in a physiological buffer. Stability. For example, 5, the region can have a Tm value, which is 1 [_1 and / or 11_/-13 in a physiological buffer! The '111 value is at least 2, 4, 6'7, or 8. 〇, for example, at a concentration of 0.5 mg/ml (mg/ml), the Tm value of a few _1 and/or IL-Ιβ is about 5~12, 5~10, or 7_1〇〇. The temperature of c. The protein may include other features described in this case. On the other hand, the feature disclosed in the present invention is that the isolated protein comprises an interleukin region of the IL-type family. Examples of members of the IL] interleukin family include IL-lct, IL-Ιβ, IL-1Ra, il-18, IL1F5, IL_1F6, IL-1F7, IL-1F8 ' IL-1F9 ' IL_1F1〇, hunger _33. The interleukin region may comprise a receptor binding region of the aforementioned interleukins, or a protein sequence comprising - or a plurality of components of these interleukins. For example, the interleukin region can comprise one or more chimeras of one of the IL-1 interleukin family members. In one embodiment, the mating region includes at least one segment having at least 5, 6, 7, 8, 9, 10, 15, 20, 25 3 , 35 , 40 , 45 , 50 , 55 ' 60 ' 65' or 70 amino acid lengths, and having an amino acid identity for a first-Μ family of interleukins (or at least 8〇, Μ, 85, 87, 15 201217528 90, 92, 94, 95, 96, 97,98, or 99% identical), and another segment having at least 5,6,7,8,9,10,15,20 '25,30, 35, or 40 amino acid lengths, And having an amino acid identity for a second IL-1 family of interleukins (or at least 80, 82, 85, 87, 90, 92 '94 '95, 96, 97, 98, or 99% identical) . Typically, the chimeric region is less than 90, 85, 80, or 75% identical to one or both of the first and second IL-1 family interleukins. In one embodiment, the first and second IL-1 family interleukins are selected from the group consisting of IL-Ιβ, IL-la and IL-IRa. In another embodiment, the first and second IL-1 family interleukins are selected from the group consisting of IL-1F5, IL-1F6, IL-1F7, and IL-1F8. In another embodiment, the first IL-1 family of interleukins is selected from the group of agonists and the second IL-1 family of interleukins is selected from the group of antagonists. In some embodiments, the chimeric region comprises less than 120, 110, 100, 90, or 80 conjugated amino acids from the same parenteral region. In one embodiment, the chimeric region is at least 50, 60, 70, 80, 90, 100, 110 or 120 positions identical to the first IL-1 family of interleukins and at least 50, 60, 70, 80, 90, 100, 110 or 120 positions are identical to the second IL-1 family of interleukins (including positions that can be individually identical to the two interleukins). In one embodiment, the chimeric region comprises at least 2, 3, or 4 discrete segments, each segment having at least 5, 6, 7, 8, 9, 10, 15 or 20 amino acid lengths, and The amino acid identity is identical for a corresponding segment of a first IL-1 family of interleukins (or at least 80, 82, 85, 87, 90, 92, 94, 95, 96, 16 201217528 97, 98, or 99%) The same), and in the remaining positions, includes amino acids mainly derived from a second IL-1 family of interleukins. In one embodiment, the chimeric region comprises 4' 5, 6, or 7 segments, wherein adjacent segments are derived from different parent IL-1 family interleukin regions. For example, the various amino acid groups in this region are located on a peptide chain from a naturally occurring human IL-1 family of interleukin regions, at least 5 or 6 amino acid lengths. In one embodiment, the chimeric region comprises at least one of the following options, two or three: (i) from IL-Ιβ, at least 50, 60, 65, 70, or 75 amino acid lengths One stage, (Π) from IL-IRa, at least 15, 2〇, one of 25 amino acid lengths; and (iii) from IL-IRa, at least 15, 20, 25 amino acid lengths One paragraph. In one embodiment, the discontinuous segments include (1) 1st to 6th and 45th to 61th, (ii) 1st to 6th and 86th to 95th, (iii) 45th to 61th and 86th to 95th, (iv) 1st ~ 6 with 148~153 '(v) 45~61 with 148~153, or (Vi) 86th to 95th with 148~153 residues, according to the number of these positions on iL_ip. The three discrete segments from the first IL-1 family of interleukins may include, for example, 1st to 8' 42 to 120, with 141 to 153 residues, 1 to 10, 37 to 125, and 131 to 153 residues, or 1 to 6, 45 to 61, 86 to 95, with 148 to 153 residues, according to the number of these positions on IL-Ιβ. The barrier region may be at least 80, 82, 85, 87, 90, 92, 94, 95, 96, 97, 98, 99 or 100 °/❶ at the remaining position identical to the second IL-1 family of cells. Prime area. In one embodiment, the one or more borders of the discontinuous segments are located at the same or retained (eQnserved) position of the first and second IL-1 family interleukins. The protein may have other features described in this context. 17 201217528 In another aspect, the present invention provides an isolated protein comprising an amino acid sequence of at least 80, 82, 85, 87, 88, 89, 90, 91, 92, 93, 94, 95 '96 , 97, 98, 99 or 100% identical to one of the sequences disclosed herein 'eg one of the sequences listed in Table 3 / Example 1, eg P01, P02 'P03, P04, P05, P06, or P07 The acid sequence is either a sequence contained in Example 5'6 or elsewhere in the case. The present invention also provides an isolated protein comprising an amino acid sequence from the N-terminus of thiocyanate to P (U, p〇2, p〇3, P04, P05, P06, or P07, or in Examples 5, 6 Or one of the sequences contained in the other case, and the amino acid sequence which provides the isolated protein 'including P〇p〇2, p〇3, p〇4, P05, P06' or P07, or in Example 5 or A sequence in which the N-terminal alanine is not present in the present case, the sequence of which may include other features disclosed in the present invention. For example, the sequence may further comprise a tag, such as A hexa-histidine sequence, for example, at the end of the 沁 or 匕 relative to the IL-1RI binding sequence. The sequence may further comprise a moiety that modifies the stability or pharmacokinetics of the il_1ri binding sequence. For the sequence, it may further comprise a serum albumin and/or an Fc region, or one or more regions of the above region, such as one or more immunoglobulins. & Immunoglobulin constant domains), or a gray more eve a 4 eve · eve < albumin region. White may have other features described in this context. In some embodiments, the isolated protein system, the sequence or chimeric region disclosed in the present disclosure = consists or is substantially as disclosed in the present disclosure - sequence or chimeric region 4 Composition. In another aspect, the present invention provides an isolated protein comprising a domain having the cytoplasmic region of the present invention = 18 201217528 (circularly permuted form) ' For example: listed in Table 3 One of the interleukin regions and/or one of the regions of sequence identification number No. 3 (SEq ID N〇: 3). The β-myxoid protein can be included in the Ν- or finalization of the permuted form. A heterologous sequence (e.g., an Fc region or albumin) can alternatively be separated by a linker. The protein can have other features described in this context. A pharmaceutical composition comprising one or more of the receptor binding agents described herein (eg, a protein comprising a chimeric interleukin region). The pharmaceutical composition can be an ophthalmic pharmaceutical composition, a topical composition, Or use Composition for parenteral administration On the other hand, the present invention is characterized by a method of modulating a subject's immune or inflammatory response. The method may comprise an immune or inflammatory response in an effective modulating individual. The amount 'is a composition comprising the receptor binding agent described in the present invention to one body. On the other hand, 'the present invention is characterized by treating a disease of IL-1 mediated disorder of a body. method. The method comprises administering to a subject a composition comprising a protein that binds to one of IL-1RI (e.g., one of the receptor binding agents described herein). For example, the disease can be an autoimmune disease, such as rheumatoid arthritis or juvenile chronic arthritis, scleroderma 'Sugeer's syndrome ( Sjogren's syndrome), ankylosing spondylitis, Behcet's syndrome, inflammatory bowel disease, asthma, vasculitis, or psoriasis 19 201217528 (psoriasis). The disease may be a disease associated with aggregate formation, such as: hyperuricemia, gout 'diabetes' (including non-insulin dependent diabetes), Alzheimer's disease Alzheimer's disease, followed by secondary reactive amyloidosis, amyotrophic lateral sclerosis (ALS), Huntington's disease, or Parkinson's disease Parkinson, s disease. The disease can also be a CAPS (CIAS1 Associated Periodic Syn syndrome) or other disease described in this case. On the other hand, the present invention is characterized by a method of treating an eye disease in a body. The method can comprise administering to a subject a composition comprising a protein that binds to one of IL-1RI (e.g., one of the receptor binding agents described herein). For example, the composition is an ophthalmic composition that can be administered topically to one of the individual's eyes or surrounding area. In one embodiment, the disease is a dry eye syndrome. In some embodiments, the individual does not exhibit the manifestations of systemic autoimmune disease. In some embodiments, the β-myxious individual has Sjogren's Syndrome (sj6gren's Syndr〇me). In some embodiments, the individual has a graft-versus-host disease (GVHD). In other embodiments, the disease is uveitis. In other respects, the present invention is characterized by a method of inhibiting the activity of babies. The method comprises contacting a receptor binding agent that binds to IL_1RI with a cell that reacts with 20 201217528 IL-1 or with a body. In general, the protein is provided in a dose effective to inhibit cell-related sputum activity or to inhibit IL-1 activity in the individual. This protein can be used to ex vivo contact with cells from one body. In another aspect, the invention features an isolated nucleic acid comprising one or more nucleic acids encoding the proteins described herein or a nucleic acid disclosed in the disclosure (eg, in Table 4), a sequence of nucleic acid hybrids, or at least 8 〇, 82, 85, 87, 88, 89, 90, 9 卜 92 ' 93 ' 94 ' 95, 96 ' 97 ' 98, 99 or 100% identical to The sequence of this nucleic acid. Exemplary hybrid sequences can be at least 2, 3, 400, 420, or 450 nucleotides in length, for example, ranging from 42 to 48 centimeters in length. The nucleic acid may also include other features disclosed in the present disclosure. The present invention also features a recombinant host cell comprising a nucleic acid comprising one or more sequences encoding the polypeptides described herein and polypeptide chains thereof. A receptor binding agent can be made by a method which comprises maintaining the δHai primary cells under conditions which permit the receptor binding agent, and optionally recovering the receptor binding. The agent, for example, is recovered from cells and media associated with the host cell. For example, the receptor binding agent can be purified from the lysate of the cell. The purified receptor binding agent can be formulated with, for example, an excipient, a stabilizer, and one or more of a buffer. The present invention is also characterized by a method of providing a chimeric protein region. The method includes identifying at least two mother 21 201217528 proteins (eg, a first parent protein and a second mother white) having a common fold, and locating at least two of the first parent proteins And a constructing a nucleic acid having a sequence encoding a chimeric amino acid sequence comprising the two segments from the first parent protein and Residues mainly from the remaining positions of the second parent protein. This region may be a region consisting mostly of β-sheets, or a region consisting mostly of α-helices, or one having these elements. The area of the combination. For example, the region can have a fold of a single interleukin. The first and second parent proteins may be related by homology, for example, the degree of amino acidity of 1 〇 to 4% by weight. In some embodiments, the segments from the first protein are located in a single folded protein region, and the chimeric amino acid sequence comprises one type of the folded protein region, ie, the lines are different Corresponding regions of the first and second parent proteins. In some embodiments, the two parent proteins have different functional properties, and the chimeric region can have one or both of the properties of the parent proteins. In some embodiments, the chimeric region has a binding interface from the first parent protein and another binding interface from the second parent protein. Reference herein is made to various regions, segments, and residues of the IL-1 family of interleukins for sites a, B, (: and 1). The residues, the location of the segment and the region are located in the sequence of human il-1β (sequence identification number i (SEQ ID NO. 1)) and the corresponding position (ρ〇3^〇η3) ) is provided below and in Figure 1: Part A. The eight residues on the IL-Ιβ include: ARGU, SERn ' 22 201217528 GLN14, GLN15, SER21, GLU25, LYS27, LEU29, HIS30, LEU31, GLN32, GLY33, GLN34, ASP35, MET36, GLU128, ASN129, and MET130 'and the corresponding residues of other IL1 interleukin family members (herein referred to as "site A residues"). In some cases, particularly in relation to IL-1 β, the reference to "extended Site A residues" includes Site A residues and GLN149, and PHE150, and others. Corresponding residues of members of the il 1 interleukin family. Furthermore, it is possible to define one of the "Site A regions" as shown in Fig. 4, including, for example, an A1 segment (corresponding to IL-Ιβ sequence identification number No. 1 (SEQ ID N0) :1) The 11th to 36th positions) and the A2 segment (corresponding to the IL-Ιβ sequence identification number No. 1 (SEQ ID NO: 1), positions 125 to 131), and other IL1 interleukin family members Corresponding paragraphs. Part B. Site B residues on IL-Ιβ include: ALAI, PR02, ARG4, GLN48, GLU51, ASN53, ILE56, LYS92, LYS93, LYS94 'LSY103 'GLU105' with ASN108, and other IL1 Corresponding residues of members of the interleukin family (herein referred to as "site B residues,"). In some cases, particularly in the case of IL-Ιβ, the reference to "extended Site A residues" includes the site B residue and the site B in Figure 4 PHE46 and SER 152 outside the zone. In addition, it is possible to define a "part β zone" as shown in Fig. 4, including, for example, a B1 segment (corresponding to the serial identification number of IL-Ιβ) N〇: 1) The 1st to 5th positions) 'A B2 segment (corresponding to the IL-Ιβ sequence identification number ID NO: 1), 48th to 56th positions), and a B3 segment (corresponding to IL-Ιβ) Sequence identification number No. 1 (SEQ ID ΝΟ: 1) positions 92 to 98), and corresponding segments on other ili 23 201217528 interleukin family members. Site C. Site on IL-1 β C residues include: ile 104, ILE106, ASN107, LYS109, GLUm, THR137, LYS138, GLY139, GLY140, GLN141, THR144, and corresponding residues of ASP145, and other members of the IL1 interleukin family (herein referred to as " Part c residue "). In addition, one of the "part C zones, which is possible" as defined in Fig. 4 is defined, including, for example, a C1 The segment (corresponding to the position 136 to 145 of the sequence identification number No. 1 (SEQ ID ΝΟ: 1) of IL-Ιβ), and the corresponding segment on other members of the IU interleukin family. Part D. Site D residues on IL-Ιβ include: LEU6, THR9, LYS63, GLU64, LYS65, and ASN66, and other ili interleukin family members corresponding residues (herein referred to as "site D residues") . Further, it is possible to define a "part D zone" as shown in Fig. 4, including, for example, a D1 segment (corresponding to IL-Ιβ sequence identification number No. 1 (SEQ ID N〇: l) 6 to 9 positions), a D2 segment (corresponding to the IL-1 β sequence identification number 丨 (SEQ ID ΝΟ: 1) positions 37 to 41), and a D3 segment (corresponding to the sequence of IL-Ιβ) Identification No. 1 (SEQ ID NO: 1), positions 63 to 66) 'A D4 segment (positions 86 to 91 of the sequence identification number No. 1 (SEQ ID NO: 1) of the deer in IL-Ιβ), And a D5 segment (corresponding to the IL-Ιβ sequence identification number 丨 ^ ^ ^ m position 150~153 position) 'and corresponding segments on other IL1 interleukin family members. Further identification of the location of the residual panel of the regions a, B, C and D can be obtained by sequence alignment of the mentioned interleukins with the sequence shown in Figure 4. .

S 24 201217528 The calculation of "homology" or "sequence identity" between two sequences (used interchangeably in the context of the present invention) is carried out in the following manner. The sequences are aligned according to the sequence alignment provided in the present case, or, in the absence of proper sequence alignment, the best sequence alignment is utilized by the Needleman and Wunsch algorithm (Needleman and Wunsch algorithm). Determined by the highest score obtained, it is separated by the Blosum 62 scoring matrix in the Needle algorithm of the European Molecular Biology Open Software Suite EMBOSS package. The gap penalty is 10, and is implemented with a parameter with a gap extend penalty of 1. See Needleman, S_B. and Wunsch, C. D. '1970, J. Mol_ Biol, 48, 443-453; Kruskal, J. B., 1983. A review of sequence comparisons can be found in Time Warps, string edits and macromolecules: the "Time Warps, String Edits and Macromolecules: the "Time Warps, String Edits and Macromolecules: the Thesis" by D. Sankoff and JB Kruskal. Theory and practice of sequence comparison), pages 1~44, published by Addison Wesley, and available from the European Bioinformatics Institute (Cambridge, UK) Μ BOSS: European Molecular Biology Open Software Kit (2000) ' Rice, P. et al., Journal of Genetics Trends, 16, (6), pp. 276-277 (EMBOSS: The European Molecular Biology Open Software Suite (2000), Rice, P. et al. , A., Trends in Genetics 16, (6) pp· 276--277), and can be found online at website 25 201217528 http://www.ebi.ac.uk/Tools/emboss/align/index.html http://emboss.open-bio.Org/wiki/Appdoc: Made at Needle. The amino acid residues or the nucleotide acid groups at the corresponding amino acid positions or nucleotide positions are then aligned. When a position on a first sequence and a corresponding position on a second sequence are occupied by the same amino acid residue or nucleotide, then the molecule is identical at that position (here used) An amino acid or nucleic acid "identity" is equivalent to an amino acid or nucleic acid "homology"). The percent identity between the two sequences is a function of the number of identical positions shared by the sequences. Determining a collective identity between a sequence of interest and a population of reference sequences, if at least one amine group at a position corresponding to one of any one or more of the group of reference sequences For acid, the position is considered to be the same. The sequence of the Provided herein is at least 80, 82, 85, 87, 88, 89, 90, 91 '92, 93, 94, 95, 96, 97, 98 or 99% identical to the sequence disclosed in the present disclosure. As used herein, the term "corresponding to" is used to indicate the position of the amino acid residue on a polypeptide of interest relative to a reference polypeptide. Generally speaking, the position Is the position specified by one of the sequence ratios specified in the present case (for example, Figure 4). Where the percentage of the same degree has been determined and no corresponding residue exists under a specific alignment, the residue The term "hybridizes under high stringency conditions" means the conditions used for hybridization and cleaning. Conducting a heterozygous reaction

S 26 201217528 Can be found in the "Current Protocols in Molecular Biology" by John Wiley & Sons, Ν·Υ·Published, 1989, 6.3.1-6.3.6 The section 'its content is incorporated herein by reference. Both aqueous and non-aqueous methods are described in this reference and can be used. High stringency conditions include heterozygous in 6X SSC at about 45 °C followed by 0. 2 Χ SSC, 0.1 at 65 °C. /. One or more cleanings are performed in the SDS, or under substantially similar conditions. The subject matter specified in the present invention is an isolated nucleic acid comprising a sequence which can be hybridized under high stringency conditions to a nucleic acid encoding the amino acid sequence disclosed in the present invention, and heterozygous to the present case (for example: Example 1) The disclosed nucleic acid. The naturally occurring protein proteins referred to in this case specifically include the human form of the protein, as well as forms from other mammalian species. All of the patent applications and references cited in this application are incorporated by reference for all purposes. BRIEF DESCRIPTION OF THE DRAWINGS Fig. 1 is a P04 structure diagram determined by X-ray crystals (X_ray cryStau〇graphic) data. The backbone from the IL_1Ra residue is shown in black, and the backbone from the IL-Ιβ residue is shown in gray. Figure 2 depicts three visual fields of a model in which the p〇5 protein binds to the extracellular region of human IL-1RI. Figure 3 depicts a model of a chimeric protein in which the IL_1Ra residue is depicted as black and the IL-lp residue is depicted as white. The proteins depicted in this model: p〇1 (Fig. 3A), p〇3 (3B@I), p〇4 (3CS|), P05 (Fig. 3D), p〇7^3E®)^ P06^3Fi|). 27 201217528 Figure 4 provides a sequence alignment of several human 1L_1 home interleukins: IL-Ιβ (SEQ ID NO: 1 (SEQ ID NO: 1)), IL-la (SEQ ID NO: 2 (SEQ ID NO) :2)) 'IL-IRa (SEQ ID NO: 3) ' IL-33 (SEQ ID NO: 4), IL-36Ra (SEQ ID NO: 4) No. 5 (SEQ ID NO: 5)) 'IL-30ot (SEQ ID NO: 6), IL_36P (SEQ ID NO: 7), and IL-36Y (SEQ ID NO: 8). The segments mentioned in the text of this case are identified in the sequence alignment. In addition, the β-folding and the loops between the flaps are also identified. The fifth panel is a list of one of the amino acid sequences of Ρ01 (SEQ ID NO: 17 (SEQ ID ΝΟ: 17)). Figure 5 is a list of one of the amino acid sequences of Ρ02 (SEQ ID NO: 18). Figure 5C is a list of the amino acid sequences of P03 (SEQ ID NO: 19). Figure 5D is a list of P04 (SEQ ID NO: 20) amino acid sequence. Figure 5E is P05 (SEQ ID NO: 21) A list of amino acid sequences. The segment from IL-Ιβ is shown in bold italics. See also Example 1 below. Figure 6 is an image of an SDS-PAGE colloid showing self-expression An exemplary sample of the protein purified from the body-binding agent of E. coli cells. The 15 and 20 kDa molecular weight marker lines are shown on the left side. The lanes are as follows: molecular weight marker (p.丨 and 6th lanes), extracts (2nd and 7th lanes), materials purified by cation exchange chromatography (3rd and 8th lanes), and anion exchange chromatography

28 201217528 The material purified by the method (the 4th and 9th lanes), and the reduced sample of the material (the 5th and 1st lanes, the 2nd to 5th lanes are P05 purified, and the 6~10 is the purified P04. See also Example 2. Figure 7A is a table with attached bar graph showing p〇6, P07 and P01 proteins, relative to IL-Ιβ and a negative control. The group (β-glucuronidase (GUS) protein), the ability to promote signal transmission. Figure 7 is a graph depicting the antagonism of IL-ip caused by Ρ01 at different concentrations of IL-Ιβ. Figure 8A is depicted in the presence of 毫. 1 ng/ml (ng/ml) of IL-1 β (human), by Ρ03 (by hexa-histidine tagged) 'P04 (via hexa-histidine marker), p〇5 (via hexa-histidine marker), and IL-IRa antagonism of IL-Ιβ. Figure 8B is a picture • .1 ng/ml (ng/ml) of IL-Ιβ (human) in the presence of an unlabeled (untagged) form of P01, P〇2, P03, P04, and P05 and IL-IRa Antagonism of IL-Ιβ caused by the product A plot of the estimated use of protein concentration in each lysate. Figure 9 is a graph containing SPR data showing the binding kinetics of immobilized soluble IL-1RI with the following proteins (binding) Kinetics) : IL-Ιβ (Fig. 9A), IL-IRa (Fig. 9B) 'P04 (Fig. 9C), and p〇5 (Fig. 9D). Fig. 10A is depicted in Example 7. The thermal denaturation of IL-IRa, IL-Ιβ, P03, P04 and P05. Figure 10B is the negative first derivative of Figure 10A. Figure 11A shows the average The bar graph of the mean fractional standard error (SEM) of the cornea is from the third, third, seventh, ninth, and eleventh days, and the application of 29 201217528 in the dry eye model of mice, two The results of corneal fluorescent staining for each eye under independent study. The number of patients who did not receive any treatment in these mice was 18 (n = 18), and received 1 mg/ml (mg/mi) p〇5. The number is 19 (n=19), or accept i.^xpBs, ie the number of vehicle handlers is (η = 20). The asterisk indicates the statistics of P05 relative to the vehicle. Significant, as follows: * (Ρ < 〇_〇5) and ** (ρ < 〇05 square). Figure 11 is a bar graph showing data from a separate experiment showing the mean corneal staining score of the cornea of each eye. The mean standard error (SEM) is based on the third, 3, 7, 9, And 11 days, the dry eye model was applied to mice. The number of patients who did not receive any treatment was 8 (η = 8); the number of patients receiving ΐ·25χ PBS vehicle was 8 (η=8), and 10 mg/ml (mg/ml) of mouse serum was received. The number of mice treated with murine serurn aibumin (MSA) was 8 (η = 8), or the number of patients receiving 1 mg/ml (mg/ml) of P05 was 9 (n=9). The asterisk indicates the statistical significance of p〇5 relative to murine serum albumin as follows: * (P < 〇·〇5) and ** (p < 0 〇〇 5). Figure 11C is a bar graph included in the same experiment as in Figure iiB, data obtained from treatment with Restasis® (0.050/cyclosporine emulsion). The asterisk indicates the statistical significance of p05 relative to Restasis® as follows: * (p < 〇.〇5) and ** (p < 0.005). Fig. 12A depicts the structure of the X-ray crystal structure (black) of P04 applied to a computational model (gray) of its structure. Figure 12B illustrates the interaction between K64 and E39 of p〇4; Figure 12C illustrates the interaction between K40 and R9 of P04 and C-terminal residues Q149 and S152. 30 201217528 [Embodiment 2] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT The IL-1 interleukin family includes several members, all having a total of s, a β-trilobal composed of six beta strands (p_tref〇il) The folding step is the same as the β-barrel structure, covered by another six beta strands. These finely divided humans are known to be the major structural lines of other mammalian forms. The phenotype 1 shows an exemplary structural arrangement of one of several human IL-1 family members. 1 Figure We found that the IL-i fold is highly plastic. The components from different members can be combined to provide a protein that promotes or antagonizes the transmission of interleukin signaling. Examples of such proteins include chimeric cell mediators = regions which include, by way of example, in the context of the presence of another interleukin or interleukin = consensus sequence, from the second or More segments or surface residues create a combination of receptor interaction sites from non-naturally occurring interleukins of different rivers. The IL-1 family of interleukins can include at least two major receptor interaction sites, termed Site A and Site B. Site a and site b are involved in contact with a major interleukin receptor (e.g., IL_1RI). It is more than _1 ft 3 and 1] 1_1 (3, for example, the two proteins are substantially different in relation to the site 8 and the site B, resulting in less contact of the IL-IRa at the site B. We found It is possible to construct a functional chimeric interleukin region comprising a site A derived from a single interleukin and a site B derived from another interleukin. The plasticity of the IL-1 fold allows for a variety of fungal cells The construction of the interleukin region is antagonized by IL-1 signaling. 31 201217528 In addition, the IL-1 family of interleukins may include two second receptor interactions (M, called site C and site D, etc.) Involving agonism and/or antagonism, and may determine the ability of the interleukin to interact with its second interleukin receptor (eg, IL_1RAcP). It is derived from a natural receptor antagonist (eg, IL_1Ra). Site c and/or site 〇 residues, which can confer antagonism. In addition, a chimeric cytokine region that antagonizes signal transduction and includes site B residues from an IL-1 agonist is Possible. The chimeric intercellular region can be constructed to include one or more ILs. -1 one or both of the agonist (eg, IL-1 β and IL-1 α) and/or the Α 丨 抬 receptor antagonist (eg, IL-IRa) One or both of site c and site d of the receptor antagonist (eg, IL-1Ra). The exemplified combinations are also provided in Table 1 below: Table 1 A Β CD 1. IL-Ιβ or ια IL- Ιβ or Ια IL-IRa IL-Ιβ or Ια 2. IL-Ιβ or ια IL-Ιβ or Ια IL-Ιβ or Ια IL-IRa 3. IL-Ιβ or ία IL-Ιβ or Ια IL-IRa IL-IRa 4 IL-IRa IL-Ιβ or let IL-IRa IL-Ιβ or la 5. IL-IRa IL-Ιβ or la IL-Ιβ or la IL-IRa 6. IL-IRa IL-Ιβ or la IL-IRa IL- The IRa s genus source sequence may be identical to the human sequence of the aforementioned interleukin region or may comprise a mutation relative to the human sequence, eg, to make it at least 70% '75%, 80%, 85% 90%, 95% or more are identical to the human sequences on the respective zones, for example, they may include one or more segments from each zone.

32 201217528 The source of site A residues and site B residues can be selected to maximize affinity for the primary receptor. For example, to bind to IL_1RI, the residue of site A can be derived from IL-IRa, and the residue of site B can be derived from IL_lp ^ a chimeric interleukin region can have a lower than 1 〇·8 , or a KD value of 10_1〇, for example, within 10 or 1 times the KD value of a natural receptor ligand such as: IL-Ιβ 'IL-lot, or iL-IRa under the same conditions, or Under the same conditions, the ability to bind to an il_i family receptor (eg, human IL-1RI) at a lower KD value than a natural receptor ligand (eg, iL_ip, IL-Ια, or IL-IRa) . Moreover, in certain embodiments, the chimeric interleukin region is at least one of a value lower than a parental interleukin region thereof and at least one of a maternal region faster than the chimeric interleukin region The kappa value, or a K()ff value that is slower than at least the parental interleukin region thereof, is combined. A chimeric interleukin region that binds to the IL-1 family receptor and antagonizes receptor signaling can be used as a receptor binding agent, for example, Lai treatment by the IL-1 family The disease of the towel. For example, in some embodiments, the chimeric intercellular region binds to a phantom and antagonizes IL-1 signaling. For example, it has an IC5 低于 of less than 1 〇〇, 1 〇, 卜.6, or 〇.3 nanomolar concentration (nM). In some embodiments, the interleukin region may be at least 4, 45 or 5 % identical, but less than identical, for example: less than 95, 9 〇, 85 or 8%, the same as - IL-1 family interleukin region. At the same time, the interleukin region may be at least 40, 45 or 50% identical, but less than identical, e.g., less than 95'90, 85 or 80% identical to a second IL_i family of interleukin regions. The first and second IL-1 family interleukin regions may be less than 5〇% of each other 33 201217528. For example, the first 丨-丨 family interleukin region can be an agonist (eg, IL-Ιβ or iL_ia), and the second il-1 family interleukin region can be a receptor antagonist (Example: IL_1Ra). In some embodiments, at least 8 〇, 85, 90 ' 92, 94 ' 95 ' 97, 98, 99, or 100% of the positions in the interleukin region are characterized by a position at each of the positions The amino acid present is the same as the first IL-1 family interleukin region or the same as the second IL_丨 family interleukin region (or the same, if the first and second IL-1 The family interleukin region is identical at a specific position) where 1% of the amino acid position on the interleukin region has this property 'this region is a complete chimera of the two interleukins (complete Chimera). The chimeric interleukin region can also be produced from two or more interleukins and can also have mutations relative to its parenteral (eg, at one or more specific positions in the presence of the amino acid, It is different from the corresponding amino acid on each of the mother cells). The interleukin region may have sites A, B, C and D residues from different IL-1 interleukin regions, and similarly may have regions A, B, C and D from different IL_interleukin regions. band. Part A. For example, in certain embodiments, an interleukin region comprises a residue from a receptor antagonist (eg, IL_1Ra) or an agonist, which is derived from the residue of the site A referred to above. At least 5, 10, 12, 15, 16, 17 or 18 residues, or at least 5' 10, 12 ' 15 ' 16, 17, 18, 19 or 20 of the above-mentioned extended part A residues a residue, or a conservative substitmi〇n portion of the residues, or at least 50 '65,75' 80, 90, 95, or 100% of the residues. In some real

34 201217528 ^The mid-interval region includes at least 70,75,80,85,88, 2 95 or 1% identical to a receptor antagonist (eg iL iRa) or a booster (eg. IL, or the residue of A1, A2, or A1+A2 on 1L_loO. In the second embodiment, an interleukin region includes residues, and the residues are at the site A as referred to in this case. At least 5, 10, 12, 15, 16 17 or 18 residues in the group, or at least 5, 10, 12, 15, 16, 17, or 18 of the extended site A residues referred to above a residue, or a replacement moiety of the residues' or at least 5 of the residues, is 8〇' 90 '95 ' or 100% identical to an IL1 agonist (eg: a residue). Site B. In certain embodiments, an interleukin region comprises a residue, and the parasitic residue is at least 2, 3, 5 of the residue at position B as referred to herein. 8, 9 '1〇' 11 ' 12 ' 13, 14 or 15 residues, or a conservative substitution of the residues' or at least 5 of these residues, 65, 75, 8〇, 9 〇, 95 or 100% is the same as an IL-agonist (eg IL) - Ιβ or IL-Ια residues). In some embodiments, the interleukin region comprises at least 70, 75, 80, 85 '88, 90, 92 '95 or 100% identical to an IL-1 cytokine B1, B2, B3, B1+B2, B1+B3, B2+B3, or B1+B2+B3 residues on agonists (eg IL-lp or IL-la) Site C. In certain embodiments, an interleukin region includes residues that are at least 2, 3, 4, 5, 6, 7 ' 8 ' of the site C residues referred to herein. 9, 10, 11 or 12 residues, or conservative substitutions of such residues, or at least 50, 65, 75, 80, 90 or 100% of the residues 35 201217528 identical to a receptor Antagonist (eg, IL-IRa residue). In some embodiments, the interleukin region comprises at least 5, 65, 75, 80, 90 or 100% identical to a receptor antagonist (eg, IL- Residues of the C1 segment on IRa). In certain embodiments, the interleukin region can include, for example, one or more of the following amino acids: a hydrophobic amino acid (eg, A Thiamine (Met) or isoleucine (I ie)), its lineage Corresponding to the position of THR137 corresponding to the sequence identification number No. 1 (SEQ ID NO: 1); a hydrophobicity, for example: an aliphatic amino acid (for example: valeric acid (Val) or isoleucine (lie)) , which is located at the position of GLN141 corresponding to sequence identification number 1 (SEQ ID ΝΟ: 1), and a non-acidic amino acid, such as an amino acid (eg, lysine (Lys) or Arginine (Arg), which is located at the position of ASP145 relative to SEQ ID NO: 1 (SEQ ID ΝΟ: 1); and the residues are located on other IL-1 interleukins. The location. Part D. In certain embodiments, an interleukin region residue that is at least 1, 2, 3, 4, 5 or 6 residues in the residue of the site D referred to herein, or the residues A conservatively substituted portion of the group, or at least 50, 65, 75, 80, 90, 95 or 100% of the residues, is identical to a receptor antagonist (eg, an IL-IRa residue). In some embodiments, the interleukin region comprises at least 50, 65, 75, 80, 90, 95 or 100% identical to a D1 segment on a receptor antagonist (eg, IL-IRa), D2 segment, D3 Segment, D4, D5, D1+D2, D1+D2+D3, residues in combination with these segments. Several residues on IL-Ιβ are in contact with or adjacent to IL-1RI, for your J: ALAI, PR02, VAL3, ARG4, LEU6, ARG11, SER13, GLN14, GLN15, GLU25, LYS27, LEU29,

36 201217528 HIS30, LEU31, GLN32, GLY33, GLN34, ASP35, MET36, GLN38, GLN39, PHE46, GLN48, GLU5 SER52, ASN53, LYS55, ILE56, PR057, LYS92, LYS93, LYS94, LYS103, GLU105, ASN108, ALA127, GLU128, ASN129, MET130, GLN141, GLN149, PHE150, and SER152. In addition to the designations described above at the site, these residues can be classified into two groups: Group 1 (Set 1) and Group 2 (Set 2). Exemplary Group 1 residues on IL-Ιβ include: ARG11, SER13, GLN14, GLN15, GLU25, LYS27, LEU29, HIS30, LEU3, GLN32 'GLY33, GLN34, ASP35, MET36, GLN38, GLN39, ALA127, GLU128, ASN129 , MET130, and GLN141 and corresponding residues on other IL-1 interleukin family members. An exemplary group 2 residue on m-ΐβ includes: ALAI, PR02, VAL3, ARG4, LEU6, PHE46, GLN48, GLU5, SER52, ASN53, LYS55, ILE56, PR057 'LYS92, LYS93, LYS94, LYS103, GLU105, ASN108 'GLN149, PHE150, and SER152 and corresponding residues on other IL-1 interleukin family members. In certain embodiments, an interleukin region comprising an IL-Ιβ residue comprises at least 15, 16, 17, 18, 19, 20 or 21 residues of the first set of residues referred to above. In certain embodiments, the 'interleukin region comprising the IL-Ιβ residue comprises at least 15 ' 16, 17, 18, 19, 20, 21 or 22 residues of the second group of residues referred to above. . Other variants that can be used as receptor binding agents include proteins having sequences derived from two or more members of the IL-1 interleukin family. Examples of such variants include chimeric regions based on IL-Ιβ and iL-iRa. Example 37 201217528, such variants may include one or more amino acid residues from Group 1 of IL_1Ra (eg, all Group 1 residues from IL-1Ra) and one or more from IL - Amino acid residues of Group 2 of Ιβ (for example, all Group 2 residues from iL-ΐβ). The exemplified chimeric protein is at the position of the following amino acid (for example, based on the corresponding position on the IL-1 β, the corresponding residue of the ρ sequence identification number No. 1 (SEq ID NO: 1) 1~ 8,42~120, with 141~153, sequence identification number No. 1 (SEQ ID ΝΟ:1) residues 1~6, 45~6 bu 86~95, with 148~153; with sequence identification number 1 The residues (SEQ ID ΝΟ: 1) have residues 1 to 10, 37 to 125, and 131 to 153; predominantly (for example, at least 50, 60, 70, 75, 78, 80, 82, 84, 86) , 88, 90, 92, 94, 96, 97, 98, 99 or 100%) is identical to IL-Ιβ. The remaining residues can be significant (for example: at least 50, 60, 70, 75, 78, 80 , 82 , 84 , 86 , 88 , 90 , 92 , 94 ' 96 , 97 , 98 ' 99 or 100 ° /.) is the same as IL-1Ra. For example, the following amino acid positions can be significantly the same IL-IRa: residues 9 to 41 and 121 to 140 of SEQ ID No. 1 (SEQ ID ΝΟ: 1); residues 7 to 44, 62 to 85 of SEQ ID NO: 1 , with 96~147; and sequence identification number No. 1 (SEQ 1〇>1〇:1) Residues 11 to 36 and 126 to 130. In certain embodiments, the interleukin region is identical to IL-Ιβ at positions other than IL-Ιβ at the amino acid positions Gln48 to Asn53 of IL-Ιβ, and is also at least 2,4, 5, 10 or 20 positions are identical to IL-Ιβ, and, for example, the interleukin region is at least 50, 60 '70, 75, 78, 80, 82, 84, 86, 88, 90, 92 ' 94 , 96, 97, 98 '99 or 100% identical to a cell other than IL-Ιβ

38 201217528 Interleukin. In certain embodiments, the interleukin region comprises at least 3, 4, 5, 6, 7 or 8 residues based on positions 1 to 8 corresponding to sequence identification number 1 (SEQ ID NO: 1) Same as IL-Ιβ. For example, it is included in residues corresponding to IL-Ιβ at 3, 4 or all 5 positions in ALAI 'PR02, VAL3, ARG4 and LEU6. In certain embodiments, the interleukin region comprises at least 8, 9, 10, 11, 12 ' 13 ' 14, 15, 16, or 17 residues based on the corresponding sequence number 1 (SEQ ID NO: 1 The 45th to the 61st positions are the same as IL-Ιβ. For example, it includes residues based on PHE46, GLN48, GLU51, SER52' ASN53, LYS55, ILE56, and the same as IL-Ιβ at 3, 4, 5' 6, 7, or 8 positions in PR057. In certain embodiments, the interleukin region comprises at least 5, 6, 7, 8, 9, or 10 residues based on positions 86-95 corresponding to sequence identifier number 1 (SEQ ID ΝΟ: 1) Same as IL-Ιβ. For example, it includes residues corresponding to IL-1 β at 1, 3 or all 3 positions corresponding to LYS92, LYS93 and LYS94. In certain embodiments, the interleukin region comprises at least 3, 4, 5, or 6 residues based on the 148th to 153th positions corresponding to the sequence identifier No. 1 (SEQ ID ΝΟ: 1). IL-Ιβ. For example, it includes residues based on GLN 149, ΡΗΕ 150 and SER 152 are the same as IL-1 β at all 2 or all 3 positions. In certain embodiments, the interleukin region comprises at least 5, 6, or 7 residues based on the 39th to the 36th position of the sequence identification number No. 1 (SEQ ID ΝΟ: 1) iL-IRa, for example, at least 5, 6 or 7 residues are identical to YLQGPNV (SEQ ID NO: 43). For example, the interleukin region comprises at least 10, 12, 14, 16, 18, 19, 20, 21, 22, 23, 24, 25 or 26 residues based on the corresponding sequence number 1 (SEQ ID) ΝΟ: 1) The 11th to 36th positions are the same as IL-IRa. The interleukin region may also include an allelic residue at position 145 corresponding to SEQ ID NO: 1 (SEQ ID NO: 1). In certain embodiments, the interleukin region comprises at least 5, 6, or 7 residues based on the same 30-36 positions corresponding to the sequence identifier No. 1 (SEQ ID ΝΟ: 1). IRa, for example: at least 5, 6 or 7 residues are identical to YLQGPNV (SEQ ID NO: 43). For example, the interleukin region comprises at least 10, 12, 14, 16, 18, 19, 20, 21, 22, 23, 24, 25 or 26 residues based on the corresponding sequence number 1 (SEQ ID) ΝΟ: 1) The 11th to 36th positions are the same as IL-IRa 〇 The interleukin region may also include an alkaline at the 145th position corresponding to SEQ ID NO: 1 Residues. The interleukin region may also comprise at least 4, 5, 6, or 7 residues based on the same as IL-IRa corresponding to positions 126-132, eg, at least 4, 5, 6 or 7 residues are identical to MEADQPVS (SEQ ID NO: 44). In certain embodiments, the interleukin region comprises at least 10, 12, 14, 16, 18, 19, 20, 21, 22, 23 or 24 residues, which corresponds to sequence identification number No. 1. The 62nd to 85th positions of (SEQ ID ΝΟ:1) are the same as IL-Ιβ,

S 40 201217528 In some embodiments, the interleukin region comprises one or more of the following amino acid sequences (eg, all four of the following): RSLAFR (SEQ ID NO: 45) ), IDVSFV (SEQ ID NO: 46), KKMDKR (SEQ ID NO: 47), and KFYMQF (SEQ ID NO: 48) 48)); one or more of the following amino acid sequences (eg, all four of the following): RSLAFR (SEQ ID NO: 45) ' IDVSFV (SEQ ID NO: 46 ( SEQ ID NO: 46)) 'NKLSFE (SEQ ID NO: 49 (SEQ ID ΝΟ..49)) ' and KFYMQF (SEQ ID NO: 48); in the following amino acid sequence One or more (eg, all four of the following): RSLAFR (SEQ ID NO: 45), EEKFSM (SEQ ID NO: 50), RFVFIR ( Sequence ID No. 51 (SEQ ID NO: 51)), and VTKFTM (SEQ ID NO: 52); one or more of the following amino acid sequences (eg, the following four) All): RSLAFR (sequence recognition No. 45 (SEQ ID NO: 45)) 'EEKFSM (SEQ ID NO: 50), FESAAC (SEQ ID NO: 53), and VTKFTM (SEQ ID NO: 53) Sequence ID No. 54 (SEQ ID NO: 54)); or one or more of the following amino acid sequences (eg, all four of the following): LNCRIW (SEQ ID NO: 55) )) ' EEKFSM (SEQ ID NO: 5〇) ' PNWFLC (SEQ ID NO: 56)) and KFYMQF SEQ ID NO: 48 ((SEQ ID NO: 48)). Cognac can be used for a variety of purposes. For example, it can be used for 41 201217528 to increase or decrease the activity of receptor signaling, to detect cells expressing receptors, or to purify the cells or proteins to which they bind. A specific example of a chimeric interleukin region based on IL-1 β and IL-Ra is provided in Example 1 below and includes the following exemplary antagonizing interleukin region for IL-1 signaling: P01. The P01 region comprises three segments from IL-IRa, which correspond to the amino acids Alal2~Val48, Ile60~Val83, and Asp95~Tyrl47 of SEQ ID NO: 3, and from The remaining four segments of IL-Ιβ. Overall, the P01 region has 74 amino acids (about 48% identical) from 153 amino acids of IL-Ιβ and 119 amino acids from IL-IRa (about 77°/. the same degree). These percentages add up to more than 100% because some of the amino acids of P01 and other exemplary protein lines disclosed in this case are amino acids that are conserved between IL-Ιβ and IL-IRa, thus contributing to IL-Ιβ and The percentage identity of IL-IRa is 〇P02. The P02 region includes three segments from IL-IRa, which correspond to the amino acid of the sequence identification number No. 3 (SEQ ID NO: 3), Alal2 to Val48, 146〇~¥3183, and 861'11〇~ Ding Wei 1'147, and the remaining four paragraphs from 11>-10. Overall, the P02 region has 85 amino acids (about 55% identical) from 153 amino acids of IL-Ιβ and 108 amino acids from IL-IRa (about 70% identical). P03. The P03 region includes two segments from IL-IRa, which correspond to the amino acids Alal2 to Lys45 and PhelOO to Lysl45 of SEQ ID NO: 3, and the rest from IL-Ιβ. Three sections. Overall

S 42 201217528 』 'The P03 region has 94 amino acids from about 153 amino acids (about 61% identical) and 91 amino acids from IL_1Ra (about 64% identical) degree). P04. The P04 region includes two segments from MIL_1Ra corresponding to the amino acids Alal2 to Lys45 and Alall4 to Lysl45' of Sequence Identification No. 3 (SEQ ID NO: 3) and the remaining three segments. Overall, the P04 region has 104 amino acids (about 68% identical) from 153 amino acids of several-ΐβ and 89 amino acids from _11^ (about 58% of the same). P05. The P05 region includes two segments from IL_1Ra corresponding to amino acid Argl4~Lys45 and Phel2〇~TyrlO of SEQ ID NO: 3, and the remaining three segments from IL_ip. In general, the P〇5 region has a 108-month-based base acid (about 70% identical) from 153 amino acids of IL-Ιβ and 85 amine groups from a few _11^. Acid (about 55% identical). Other examples include the foregoing examples, where the finger segments of IL-IRa bind to corresponding residues from IL-la and not to the corresponding residues from IL_ip. Protein Modifications and Substitutions Protein sequences, such as those described herein, can be modified (e.g., by manufacturing- or multiple retention substitutions) to create a retention substitution to maintain or make a modest change in function. Exemplary retention substituents are as follows: 43 201217528 Table 2 Exemplary substitutions of pro-amino acid further specific substitutions of alanine [Ala (A)] proline (val); leucine (leu); Leucine (ile) proline (val) arginine [Arg(R)] lysine (lys); glutamic acid (gin); aspartic acid (asn) lysine (lys) Aspartic acid [Asn(N)] glutamic acid (gin); histidine (his); lysine (lys); arginine (arg) citrate (gin) aspartic acid [Asp (D)] glutamic acid (glu), face acid (glu), cysteine [Cys(C)], serine (ser), serine (ser), glutamic acid [Gin (Q)] Asparagine (asn) aspartate (asn) glutamic acid [Glu (E)] aspartate (asp) aspartic acid (asp) glycine [Gly (G)] guanamine Acid (pro); alanine (ala) alanine (ala) histidine [His (Η)] aspartic acid (asn); glutamic acid (gin); lysine (lys); spermine Acid (arg) arginine (arg) isoleucine [He (1)] leucine (leu); valine (val); methionine (met); alanine (ala); Acid (phe); leucine (leu) leucine (leu) leucine [Leu (L)] norleucine (isoleucine); isoleucine (ile); Amine (val); methionine (met); alanine (ala); phenylalanine (phe) isoleucine (ile) lysine [Lys (K)] arginine (arg); Aminic acid (gln); aspartic acid (asn) arginine (arg) methionine [Met (Μ)] leucine (leu); phenylalanine (phe); isoleucine (ile) Leucine (leu) phenylalanine [Phe(F)] leucine (leu); valine (val); isoleucine (ile); alanine (ala); sulphate (tyr) white Aminic acid (leu) proline [Pro (P)] alanine (ala) alanine (ala) serine [Ser (S)] polyglycine (1) ir) physic acid (thr) polyglycine [Thr(T)] serine (ser) serine (ser) tryptophan [Trp(W)] tyrosine (tyr); phenylalanine (phe) glutamate (tyr) tyrosine [Tyr (Y)] aminic acid (trp); phenylalanine (phe); heart amine (thr); serine (ser) phenylalanine (phe) proline [Val (V)] isoleucine ( Ile); leucine (ieu); 曱 thioacetate (met); phenylalanine (phe); leucine (leu); norleucine (ala) ------- The choice of substitution can be based on the potential effects of: (a) a skeletal structure adjacent to the substitution, for example, a flap or snail The shape or the hydrophobicity of the molecule at the target site or the volume of the side chain (v〇iUme) and the branching (branching). Amino acid residues can be classified based on the nature of the side chain: (1) Hydrophobic: norieucine, methionine (met), alanine (ala), proline (vai), leucine (ieu), isoleucine (ile); (2) neutral hydrophilic: cysteine (cys), serine (ser), polyglycine (thr); aspartic acid (asn); glutamic acid (gin); (3) acidic: aspartic acid (asp), glutamic acid (glu); (4) basic (basic) : histidine (his); lysine (iys), arginine (arg); (5) residues affecting the bone matrix: glycine (gly), proline (ρΓ0); and (6) Aromatic: tryptophan (trp), tyrosine (tyr), streptoalanine (phe) ° Non-conservative substitution (Non-conservative substitution) may include one of these classes One member replaces one of the different categories into a shell. Conservative substitution may include replacing one of the members of the same category with one of one of these categories. The importance of a particular residue can also be assessed based on the hydropathic index of the amino acid. Each amino acid is given a hydrophilic index based on its hydrophobicity and electrical properties: is〇leucine (+4.5); valine (+4 2); leucine (leudne) (+3 8); phenylalanine (+2.8); cysteine (+2.5); methionine (+1.9); alanine (+) 1.8); glycine (-0.4); threonine (-0.7); serine (-0.8); tryptophan (-0.9); tyramine Acid (tyrosine) (-1.3); 45 201217528 Proline (-1.6); histidine (-3.2); glutamate (-3.5); glutamine (-3.5); aspartate (-3.5); asparagine (-3.5); lysine (-3.9); and arginine (arginine) -4.5). For a discussion of the hydrophilic amino acid index and its importance, see, for example, Kyte et al., 1982, J. Mol. Biol., 157: 105-13. The sequence of a protein can be used by any Methods to change, including oligonucleotide-mediated (site-directed mutagenesis), (Carter et al., Nucl. Acids Res_, 13:4331, 1986 Zoller et al., Nucl. Acids Res., 10:6487, 1987, cassette mutagenesis (Wells et al., Gene, 34:315, 1985) Restriction selection mutagenesis (Wells et al., Philos. Trans. R. Soc. London '317:415, 1986), and polymerase chain reaction (PCR) mutation . See also In Vitro Mutagenesis Protocols: Third Edition, Braman (eds.), Humana Press, 2010, ISBN number: 1607616513 and Polymerase Chain Reaction Selection Test Criteria (PCR Cloning) Protocol): From Molecular Cloning to Genetic Engineering, Chen and Janes (eds.), published by Humana Press, 2002, ISBN number: 0896039730. Scanning amino acid analysis can be used to evaluate one or more amine groups on a contiguous sequence

46 201217528 Acid. The method relates to mutating a single or nearly individual amino acid on a zone to a specific amino acid, for example, a mutation to a smaller, neutral amino acid such as alanine Serine or valine. Alanine is usually chosen because it eliminates side chains other than beta-carbon and is the least likely to change the backbone configuration of the variant (Cunningham and Wells, Science Journal (Science), 244: 1081-1085 '1989. It is also the most common amino acid and is often found in both buried and exposed positions. (Creighton, The Proteins, (W. H. Freeman & Co., N.Y.); Chothia, J. Mol. Biol., 150: 1, 1976). The scanning method can also be used to make more significant changes, for example, a charged residue can be changed to an oppositely charged residue, and a residue having a short side chain can be replaced with a large volume. Bulk side chains. For example, arginine scanning is a method that can be used in place of alanine scanning or as a method other than alanine scanning. The scanning method can be applied to each residue on a zone or to residues having specific properties, such as residues at or near a protein surface or residues that may be at or near the surface of the protein. The structure of a protein, one of its regions, or a complex with the protein, can be modeled, for example, by performing homology based modeling, energy Energy minimization and/or other simulations that utilize known known structures. These methods include: AMBERTM simulation software (modeling 47 201217528 software) (Case et al., 2005, Journal of Computational Chemistry (J. Computat. Chem), 26, 1668-1688 and Case et al., 2010, AMBER 11 , University of California, San Francisco, CA USA) and CHARMMTM modeling software (Molecular Simulations Inc.). For background information, see Baker and Sali, Science, 294 (5540): 93-6, 2001. This structure can also be determined directly, for example, using X-ray crystallography and/or NMR spectroscopy. Exemplary PDB structures describing the IL-1 family of interleukin structures include: 1I1B, 1ILR, 1IRA, 1ITB, 2I1B, 2ILA, 2KLL, 4I1B, 5I1B, 6I1B, 7I1B, 8I1B, 9ILB, and 1MD6 (accessible by the Internet, Available from Piscataway NJ, USA, Russell University RCSB database (RCSB-Rutgers) website www.pdb.org, and Bethesda, Maryland, USA (Bethesda, MD, USA) ), National Library of Medicine 〇 For example, the individual structure of IL-Ιβ and the complex structure formed by IL-Ιβ and receptor IL-1RI have been solved. See, for example, Finzel et al., 1989, J. Mol. Biol. 209: 779-791, ?08 103, and ¥6^ et al., 1997, Nature Journal @&^^ ) 386: 190-194. The structure in which IL-IRa binds to IL-1RI has also been solved. See, for example, PDB 1IRA and Schreuder et al., 1997, Nature, 386: 194-200. Homology modeling can be ranked by sequence

S 48 201217528 (alignment) to assist, for example, the use of computer software, such as Basic Local Alignment Search Tool (BLAST), PSI-BLAST, PHI-BLAST, WU-BLAST-2, and / or MEGABLAST. See Altschul et al., 1990, J. Mol. Biol, 215, 403-410; Altschul et al., 1996, Methods in Enzymology 266, 460-480; and Karlin et al. , 1993, Proceedings of the National Academy of Sciences (PNAS USA) 90, 5873-5787. Other algorithms for aligning macromolecules (amino acid sequences and nucleic acid sequences) include FASTA (Pearson, 1995, Protein Science '4, 1145-1160), ClustalW (Higgin et al., 1996). Methods Enzymol. (266, 383-402), DbClustal (Thompson et al., 2000, Nucl. Acids Res., 28, 2910-2926), and drug development environment software (Molecular Operating Environment), (Chemical Computing Group, Montreal, Montreal, H3A 2R7). In addition, the Myers and Miller algorithms are incorporated into the GIG sequence alignment software package (the version of Myers & Miller) (Myers & Miller, Computer BioApplications (CABIOS), 4, 11-17 , 1988) can be used. A consensus sequence between IL-1 β and IL-IRa can be obtained by comparing the two sequences and identifying the same or highly conserved residues. The region of the interferon region described herein may have at least 60, 70, 80, 90, 95 or 100% of the same residues or highly conserved residues. Other consistent sequences can be identified in the same manner. 49 201217528 Further variants of the IL-1 interleukin family members can be made and evaluated using a diSpiay_based system, for example: proteins can be displayed and assessed for binding to a tie-1 The ability of a receptor for one of the members of the interleukin family. For example, the variants of IL-1 β, IL-1 Ra or a single interleukin region described in the present invention can be evaluated for their ability to bind to a soluble extracellular domain of 11^1. . A general description of a display-based system includes the following literature: on cell display 'Chao et al., Nature-Laboratory Handbook (Nat Protoc.) '2006; 1(2):755- 68; Colby et al., Methods Enzymol., 2004; 388: 348-58; Boder et al., Methods Enzymol., 2000; 328: 430-44), on sputum Phage display (eg, Viti et al., Methods Enzymol., 2000; 326: 480-505 and Smith, 1985, Science, 228: 1315-1317), And on ribosome display technology (eg: Mattheakis et al. '1994' Proc_ Natl. Acad. Sci. USA, 91: 9022 and Hanes et al., 2000, Nat Biotechnol '18:1287-92; Hanes et al., 2000, Methods Enzymol. 328:404-30; and Schaffitzel et al., 1999, J Immunol Methods., 231(1- 2): 119-3. Although many of the examples in this case utilize the human IL-1 interleukin sequence as the mother The parental domain is used to illustrate that other sequences can be used because, in addition to the protein sequences described in this case, 'a few additional examples' are, for example, from other species, are known and available, and are Public funds 50 201217528 were found in the library, for example: Entrez (National Library of Medicine, Bethesda MD) and EBI-EMBL (Hinxton, Cambridge, UK) )). Examples of such sequences from the UNIPROT database (available from UniProt.org and refer to The UniProt Consortium, Nucleic Acids Res, D142-D148, 2010) include those shown below : Accession Numbers IL-la P01583, P01582, P16598, P08831, Q28385, Q28579, 046612, P48089, PI8430, P04822, P46647, 046613, Q3HWU1, P79161, Q60480, P79340 IL-Ιβ P01584, P10749, Q28386, P09428, P14628, P21621, Q63264, P41687, P48090, Q9YGD3, P26889, Q2MH07, Q28292, Q9WVG1, P46648, P 79182, P51493, Q865X8 IL-IRa P18510, P25085, 018999, P26890, P25086, Q9GMZ4, Q9BEH0, 077482, Q866R8, Q29056 IL-1RI Q02955, P14778, P13504 IL-lRAcP Q9NPH3, Q61730, P59822, Q63621 The cells described in this case The interleukin region may also include substitutions present on a variant interleukin region that binds to a major receptor of the IL-1 family (eg, IL-1RI). For example, the 15th position of the sequence identifier No. 1 (SEQ ID NO: 1) (corresponding to the 20th position of SEQ ID NO: 3) may be methionine (Met) or aspartic acid (Asn). The 30th position of the sequence identification number No. 1 (SEQ ID ΝΟ: 1) (corresponding to the 34th position of SEQ ID NO: 3) may be glycine (Gly), group Amine (His), tryptophan (Trp), or guanidine thiocyanate (Met). Other exemplary IL-Ιβ and IL-IRa variants are included in the following papers. 51 201217528: Evans et al., J. Biol. Chem., 270: 11477 '1995, with Greenfeder et al. Human, Journal of Biochemistry (j Biol. Chem.), 270: 22460, 1995. For example, variants of υβ include R11G, R11A, Q15H, E105G, and T147G. See, for example, the paper by Evans et al. Variants of IL-IRa include W16Y, Q20M, Q20N, Y34G, Y34H, Y34W, Y34M, Y147G, Y147H, Y147M, K145D, H54P, V18S, T108K, C116F, C122S, C122A, Y147G 'H54P, H54I' and other records In Evans et al., and Greenfeder et al., J. Biol. Chem., 270: 22460, 1995. An interleukin region can include a residue identical to IL-IRa at one of the foregoing positions. An interleukin region may also include a residue' which differs from one of the aforementioned mutations at a corresponding position. Other variants may include those that result in agglomeration or performance problems at the residue, such as W16, Y34, K93 or R97 of SEQ ID NO: 3 (SEQ ID NO: 3). For example, the one or more amino acids located at positions W16 and Y34 corresponding to SEQ ID NO: 3 (SEQ ID NO: 3) can be a non-aromatic amino acid. The amino acid at a position corresponding to the position of K93 may be an amino acid other than an acid (eg, a non-basic amino acid such as alanine or an uncharged hydrophilicity) Amino acid or an acid amino acid. The amino acid at a position corresponding to the position of R97 may be an amino acid other than arginine, for example: a non-initial amino acid such as alanine or an uncharged hydrophilicity An amino acid or an acid amino acid such as aspartic acid. 52 201217528 In addition, IL-1 family interleukins may include one or more unpaired cysteine residues. One or more 'eg two, three or all such unpaired cysteine residues can be mutated to another amino acid, eg an uncharged amino acid like propylamine Alanine or serine. For example, P01 includes cysteines at positions 67, 70, 116 of sequence identification number 17 (SEQ NO: 17) and 122 positions. One, two, three or all four of the semi-proline acids may be substituted by another amino acid, for example: an uncharged amino acid such as alanine or serine (serine) ). P02 includes cysteines at positions 67, 70, 116' and 122 positions of SEQ ID NO: 18 (SEQ ID NO: 18). One, two, three or all four of the cysteine acids may be substituted by another amino acid, for example: an uncharged amino acid such as alanine or serine (serine) ). P03 includes the 70th '116, located at SEQ ID NO: 19, and the cysteine at 122 positions. One or two or all three of the cysteine acids may be substituted by another amino acid, for example, an uncharged amino acid such as alanine or serine. P04 includes the 7th ’ ' 116 of sequence identification number 20 (SEQ ID NO: 20) and the semi-proline (CySteines) at 122 positions. One, two or all three of the semi-deaminating acids may be substituted by another amino acid, for example: an uncharged amino acid such as alanine or serine. P〇5 includes cysteines at positions 8, 70, and 122 positions of SEQ ID NO: 21 (SEQ ID NO: 21). The cysteine 53 201217528 — or all three may be replaced by another amino acid — for example: an uncharged makeup and μ , an amine I 'like alanine or serine (serine) ). The IL 1 steroidal interleukin region, including the chimeric cells described in the present invention, is also cyclically permutated. For example. 'The c-terminal segment from one of the regions can be repositioned 'with respect to the original n-terminus (N_terminus) and an N-termmal segment (generally including the c-terminal segment after excision) The remaining part of the original protein) becomes the N-terminal. The two relocated segments can be separated by a linker (e.g., having an amino acid bound to between about 3 and 10). In general, all of the amino acids in the region are preserved prior to permutation, but only in sequence. In some embodiments, the 'cutp〇int' for alignment can be placed on a curved zone, such as a flexible loop, such as β6-β7 (eg, corresponding to sequence identification) No. 3 (SEQ ID NO: 3) at positions 71 to 80 of the amino acid) or β7-β8 loop (for example, corresponding to the sequence identification number No. 3 (SEQ ID ΝΟ: 3) of the 84th~ 99-position amino acid). In some embodiments, a receptor binding agent described in the present invention, for example: a receptor binding agent comprising an IL-1 family interleukin region having less than 30, 25, 22, 20, 19, 18 ' or a molecular weight of about 17 kDa. In some embodiments, the receptor binding agent has a molecular weight greater than 18, 19, 20, 22, 25, 30 ' 40 ' 45, or 50 kDa. For example, the receptor binding agent can include other polypeptides, polymeric or non-polymeric components, such as: Pharmacokinetics, stability,

54 201217528 Immune generating power (imnum〇genicity), and / or molecular weight components. The protein may include other modifications (m〇dificati〇ns) such as post-tr-nal or synthetic modifications (symhetic (10) gamma (4) (d)). In certain embodiments, the receptor binding agent is not glycosylated. In other embodiments, the receptor binding agent comprises at least one glycosylation. For example, the receptor binding agents described herein can be modified, for example, to include other regions and features. For example, a receptor binding agent can be used in a region of an antibody protein, for example, for an Fc domain or one or more constant regions (eg, CHI ' CH2 or CH3). For example, the region can be a variant of a human region or a human region. The Fc region or the one or more constant regions may be located at the N-terminus or C-terminus of the receptor binding agent.

The Fc region may be derived from any suitable immunoglobulin, for example, from a human antibody, such as:

IgG1, IgG2 'IgG3' or IgG4 subtypes 'IgA, IgE, IgD or IgM antibody. In one example, the fc region comprises a sequence that occupies from a position about Cys226, or an amino acid residue at about pr 〇 230, to the C-terminus of the Fc region. An Fc region typically includes two constant regions 'one CH2 region and one CH3 region, and optionally a CH4 region. Antibodies having substitutions on their Fc regions and having increased serum-half-lives are also described in WO 00/42072, WO 02/060919; Shields et al., J. Biol. Biol. Chem.), 276: 6591-6604, 2001; Hinton, Biochemistry 55 201217528 (J. Biol. Chem.), 279: 6213-6216 '2004). The numbering on an IgG heavy chain is equivalent to the number of the EU index by Kabat et al., see Sequences of Proteins of Immunological Interest, Fifth Edition (5th Ed), Public Health Service, NH1, MD, 1991, number of human IgGl EU antibodies. In one embodiment, a receptor binding agent comprises an epitope that binds to a salvage antibody, which is added to an antibody (particularly an antibody fragment) to increase the antibody in vivo (in vivo) The serum half-life is as described in U.S. Patent No. 5,739,277 (U.S. Patent No. 5,739,277) and to Ghetie et al., Ann. Rev. Immunol., 18:739-766, 2000, et al. In one embodiment, a receptor binding agent comprises a serum albumin sequence that binds to an FcRn receptor, or a portion of the sequence, or a binding to serum albumin (eg, human serum albumin) The sequence of ). For example, certain peptides that bind to serum albumin may be associated with the receptor binding agent', e.g., the sequence DICLPRWGCLW (SEQ ID NO: 22). See also, Dennis et al., J. Bi〇1 Chem, 277: 35035-35043, 2002. The receptor binding agent can be modified to include a sequence that increases the size and stability of the agent', e.g., a sequence described in WO2008/155134 or W02009/023270. Such sequences can generally be biologically inactive, e.g., they do not modulate the signaling that is mediated by members of the IL-1 interleukin family. A variety of stabilizing polypeptide sequences can be used, 56 201217528 eg, with a large amount of glycine and/or serine, and other amino acids like glutamate, days The sequence of aspartate, alanine or proline. For example, the sequence can be designed to have at least 30, 40, 50, 60, 70, 80, 90 or 100% glycine and/or serine residues. In some embodiments, the length of binding of the stabilized polypeptide sequence linked to a protein can be at least 20, 25, 35, 50, 60, 70, 80, 90, 100, 120, 140, 160, 180, 200, 250 , 300, 350, 400, 500, 600, 700, 800, 900 or more than 1000 or 2000 amino acids. The stabilizing sequence can, for example, be fused to a biologically active polypeptide, for example, to the N-terminus or C-terminus of the receptor binding agent. The combination of the stabilizing sequences can result in a significant increase in the hydrodynamic radius of the conjugated protein relative to the unmodified protein, which can be utilized, for example, ultracentrifugation, size screening chromatography Size exclusion chromatography, or light scattering, to detect. In some embodiments, the stabilizing sequence comprises a minority of the following amino acids or does not comprise the following amino acids: cysteine (to avoid formation and oxidation of disulfide) 'Methylthioamide Acid (methi〇nine) (to avoid oxidation) 'asparagines and glutamine (to avoid desamidation) and aspartate (aspartate). The stabilizing sequence can be designed to include proline residues that tend to reduce the sensitivity of prote〇lytic degradation. The interaction between the binding assay-receptor binding agent and its target can be analyzed using any method of 201217528, including, for example, radio-immunoassays, cell binding assays, Surface plasmon resonance (SPR). An exemplary cell binding assay using radio-iodinated protein competition is described in the Journal of Boraschi' Immunology (J. Immunol.) 155(10): 4719-25, 1995. Surface Plasma Resonance (SPR) or Biomolecular Interaction Analysis (BIA) can detect biospecific interactions in real time without labeling any reactants. A change in mass at the bonding surface of the germanium wafer (indicating a binding event) causes a change in the refractive index of the light near the surface (the optical phenomenon of surface plasma resonance (SPR)). A change in refractivity produces a detectable signal that is measured as an indication of an immediate response between biomolecules. The method of using SPR is described, for example, in Raether, 1988, "Surface Plasmons", published by Springer Verlag; Sjolander and Urbaniczky, 1991, Analyz Chem., 63: 2338-2345; Szabo et al., 1995, Curr. Opin. Struct. Biol., 5: 699-705 and by BIAcore International AB (Uppsala, Sweden) ) Online resources provided. A BIAcore® system or a Reichert SR7000DC Dual Channel SPR (Dual Channel SPR) can be used to instantly compare interactions with rankings based on dynamics, affinity or specificity without the use of a standard

S 58 201217528 Remember. The binding affinity of a receptor binding agent to an extracellular region of an interleukin receptor (eg, the extracellular region of IL-1RI) can be close to physiological conditions, eg, 10 millimolar (mM) HEPES pH. 7.4, 150 millimolar (mM) sodium hydride (NaCl), 3 millimolar (mM) EDTA, 0.005% Tween-20, measured using SPR. Other methods that do not rely on SPR can be used, for example, to measure binding and affinity. The information obtained by the binding assay can be used to provide an equilibrium dissociation constant (KD), and kinetic parameters (e.g., KQn and Κ) for binding a receptor binding agent to a target. ^) One of the accurate and quantitative measurements. These data can be used to compare different proteins, targets, and conditions. This message can also be used to develop structure-activity relationships (SAR). For example, the kinetic and equilibrium binding parameters of the variant protein can be used to compare to a reference or parent protein parameter. Variant amino acids at a given position can be distinguished by association with specific binding parameters, such as high affinity and low Koff values. This information can be combined with structural simulations (eg, using homologous simulations, energy minimization, or structural determination by X-ray crystallography or nuclear magnetic resonance spectroscopy) to assess the affinity of the protein. It is made into a recombinant form and may include a tag suitable for purification or immobilization, for example: a FLAG marker, the myc marker 'hemagglutinin' is labeled, His is labeled 'or The Fc region is fused. The extracellular region of the receptor protein (eg, IL-1RI' and IL-lRAcP) can be utilized in sf9 cells by baculovirus by, for example, in Table 59 201217528 'eg, in bacteria or snail cells' (bac-) performance, and is made into a recombinant form. The soluble receptor protein can be immobilized in the BIA_ system, for example, by using a wafer comprising a reagent that binds to a protein marker, for example, a wafer coated with IgG specific to the Fc region or other marker. . Cell viability assay ^: The ability of a bulk binding agent to act as a receptor antagonist can be assessed, for example, in a cell-based assay. For example, a receptor binding agent is used to assess the inhibition of IL. Several exemplary assays for hunger activity are described in the literature of Boraschi et al. and include the tau cell proliferation assay, the IL-6 and IL-8 production assays, and the inhibition of calcium influx. effect. In an exemplary assay, the ability of a receptor binding agent is assessed by its ability to inhibit IL-6 released by a few mouthwashes in human fibroblasts. The inhibitory effect on the release of interleukin by the 叩-stimulation in MRC5 cells is related to the ability of the agent to inhibit the activity of IL-1 in vivo. Details of this assay are described in Dinarello et al., "Currem Protocols in Immunology", Section 6 21_6 2 7 , published by J〇hn WUey and Sons Inc. Briefly, human Mrc5 human fibroblasts (ATCC # CCL-171, Manassas VA, USA) were grown in a multi-well plate to c〇nfluency. These cells were treated with titer of the receptor binding agent (titratecj doses) and control. The cells are then contacted with a suede in the presence of a titrant and/or control.

60 201217528 g/ml (pg/ml) of IL-Ιβ. The cells of the negative control group were not stimulated by α_ιβ. The amount of IL_6 released by each group of treated cells was measured using an IL_6 EUSA kit (eg, BD Pharmingen, Franklin Lakes, NJ, USA). Can be used as a control group including a separate buffer, IL-IRa, and antibodies against IL-Ιβ. The efficacy of a receptor binding agent (Efficacy) can be assessed in vivo. An exemplary assay is described in Economides et al., Nature Med., 9: 47-52, 2003. Briefly, mice were injected intraperitoneally with a titration dose of the receptor binding agent and a control group. At 24 hours post-injection, the mice were subcutaneously injected with recombinant human IL-1 β at a dose of 1 μg/kg (called/flavor). Two hours after the injection of IL-1 β (Peak IL-6 response time), the mice were sacrificed, blood was collected and processed into serum. Serum IL-6 levels were determined by ELISA. The percent inhibition can be calculated based on the ratio of IL-6 detected in the serum of the experimental animal relative to the IL-6 detected in the control group. Other exemplary in vivo IL-1 activity assays are described in the literature of Boraschi et al. and include anorexia, hypoglycemia and neutrophilia assays. . Methods of Manufacture Receptor binding agents can be made by expression in recombinant host cells, but can also be produced by other methods such as in vitro transcription and translation and chemical synthesis. With respect to cellular expression, one or more nucleic acids encoding a receptor binding agent 61 201217528 (eg, cDNA or genomic DNA) can be inserted for cloning or for replication. In the vector. Many vectors are publicly available. The vector may, for example, be a plasmid, a cosmid, a viral genome, a phagemid, a phage genome, or Other autonomously replicating sequences. A suitable coding nucleic acid sequence can be inserted into the vector by a variety of different methods. For example, appropriate restriction endonuclease sites can be engineered (e.g., using PCR). The coding nucleic acid sequence can then be inserted into an appropriate location using digestion and ligation of the restriction enzyme. Vector components typically include an origin of replication, one or more marker genes, an enhancer element, a promoter, and one or more transcriptions. One or more of the transcription termination sequences. The receptor binding agent may be isolated or by merging (or s) or a plurality of other components, such as a signal sequence, an epitope or a purification moiety. ), with a label, is made recombinantly. The receptor binding agent can include a pro domain of the interleukin-1 family member, for example, which can then be removed by proteolytic processing. Regarding bacterial expression, the receptor binding agent can

62 201217528 is made with or without a signal sequence. For example, it can be made in a cell to accumulate in inclusion bodies, or in a soluble fraction. It can also be, for example, by adding a prokaryotic signal sequence, such as, for example, from an alkaline phosphatase, penicillinase, or a thermostable enterotoxin II. A suitable leader sequence is secreted. Exemplary bacterial host cells for expression include any transformable E. coli K-12 strain (such as E. coli BL2, C600, ATCC 23724; E. coli HB101 NRRLB-11371). , ATCC-33694; Escherichia coli MM294 ATCC-33625; Escherichia coli W3110 ATCC-27325), B. subtilis, Pseudomonas 'and other bacilli strains. Proteins produced in bacterial systems typically lack saccharification. Thus, in some embodiments, the receptor binding agents described herein are substantially free of saccharification, for example, without the biliary modification of mammals or other eukaryotic cells. The receptor binding agent can be expressed in a yeast host cell, for example: Saccharomyces cerevisiae, Schizosaccharomyces pombe, Hanseula, or Pichia pastoris ( Pichia pastoris). Regarding yeast performance, the receptor binding agent can also be produced intracellularly or by secretion [for example, using the yeast initiase ieader or alpha factor leader (including gingival fungi) (Saccharomyces) and Kluyveromyces cerevisiae 63 201217528 (Kluyveromyces) type) or acid phosphatase leader (add ph〇sphatase leader), or C. albicans glucoamylase lead (see bulletin 丨European Patent EP 362,179, April 4, 1999. In mammalian cell expression, mammalian signal sequences can be used for direct secretion of proteins, such as secreted polypeptides from the same or related species, and viral secretory leaders. Alternatively, the receptor binding agent can be made to have a anterior region of a member of the interleukin-1 family, for example: an IL-la or IL-Ιβ pre-region. Both the expression and the selection vector comprise a nucleic acid sequence which enables the vector to replicate in one or more selected host cells. Such sequences for use in a variety of bacteria, yeasts and viruses are well known. The origin of origin from the plastid pBR322 (origin 〇f repiicati〇n) is suitable for most Gram-negative bacteria; the 2μ plastid origin is suitable for yeast; and a variety of viral origins (SV40, Polyomavirus (p〇iyoma), adenovirus, VSV or BPV are effective for colonizing vector lines in mammalian cells. The performance and selection vectors typically comprise a selecti ri gene or a marker. Typical selection genes encode proteins with the following properties. (a) Provide resistance to antibiotics (antibi〇tics) or to other toxins, eg, ampicillin, neomycin, amine Fate (methotrexate), or tetracycline (b), (b) deficiency of nutritional supplements (aux〇tr〇phic deficiencies) (eg, URA3 logo on Saccharomyces), or ( c) Supply of epical nutrients not available in complex 64 201217528 complex media, for example, a gene encoding D-alanine racemase for Bacillus. A variety of standards are also available for mammalian cells, such as dihydrofolate reduction (DHFR) or thymidine kinase. DHFR can be used in combination with a cell line lacking DHFR activity (such as a CHO cell line), and its preparation and propagation is described in Urlaub et al., Proc. Natl. Acad. Sci. USA. 77:4216, in the 1980 paper. The expression and selection vectors typically comprise a promoter operably linked to the nucleic acid sequence encoding the receptor binding agent to direct the synthesis of the mRNA. Exemplary promoters suitable for use in prokaryotic hosts include beta-lactamase and lactose promoter systems (Chang et al., Nature, 275:615, 1978; Goeddel Et al., Nature, 281:544, 1979) 'alkaline phosphatase, tryptophan (trp) promoter system (Goeddel, Nucleic Acids Res.) , 8:4057, 1980; European Patent No. 36,776 (EP 36,776)), with a hybrid promoter, such as the tac promoter (deBoer et al., Proc. Natl. Aead Sci. USA), 80: 21-25, 1983). The promoter for the bacterial system may also comprise a suitably positioned Shine-Dalgarno sequence. The T7 polymerase system can also be used to drive the expression of a nucleic acid coding sequence placed under the control of the T7 promoter. See, for example, the pET vector (EMD Chemical Company, Gibbstmvn 65 201217528 City, New Jersey, USA) (EMD Chemicals, Gibbstown NJ, USA) and host cells, such as the Novagen User Protocol TB053 provided by EMD Chemical Company and the United States Patent No. 5,693,489 (US 5,693,489). For example, the vectors can be used with 81^21 (0£3) cells and BL21 (DE3) pLysS cells to produce a protein, for example, at least 0.05 per ml of cell culture. 0.1 or 0.3 mg (mg). Other cell lines that can be used include the DE3 lysogen strain of B834, BLR, HMS174, NovaBlue, including cells having a pLysS plastid. Exemplary promoters for yeast cells include for 3-phosphoglycerate kinase (Hitzeman et al., J. Biol. Chem_ '255:2073 '1980) or other sugars. Glycolytic enzymes (Hess et al., J. Adv. Enzyme Reg. ' 7:149 '1968; Holland, Biochemistry, 17:4900, 1978), like Enolase (eno!ase), glyceraldehyde-3-phosphate dehydrogenase 'hexokinase, pyruvate decarboxylase, phosphofructokinase ), glucose-6-phosphate isomerase, and the promoter of pyruvate kinase. Other exemplary yeast promoter lines are advantageous in inducible and have other transcriptional control under growth conditions. Examples of inducible promoters include for alcohol dehydrogenase 2, isocytochrome C, acid phosphate

66 201217528 acid phosphatase ' metallothionein, which is encoded in the promoter region of the mammalian host cell with the promoter regions responsible for the enzymes used by maltose and galactose The expression of mRNA of a receptor binding agent can be controlled, for example, by a promoter derived from a viral genome, such as polyoma virus, adenovirus (like Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, retrovirus 'hepatitis B Hepatitis-B virus and Simian Virus 40 (SV40), derived from heterologous mammalian promoters, such as the actin promoter, or an immunoglobulin ( The immunoglobulin promoter is associated with a promoter derived from heat-shock. A heterologous promoter system can also be used, for example, a promoter that is responsive to tetracycline. See Urlinger, s et al., 2000, Proc. Natal. (USA) 97 (H): 7963-7968. Transcription can also be driven by an enhancer sequence located in the cis or trans. Exemplary mammalian enhancer sequences include those used for globin, elastase, albumin, alpha-fetoprotein (a_fet〇pr〇tein), and insulin (insulin) enhancers. sequence. Other examples include the SV40 enhancer (bp 100-270) located on the late side of the replication origin. The cytomegalovirus early promoter recognizes 67 201217528, located near the origin of replication. Tumor enhancer (p〇ly〇ma enhancer), with adenovirus enhancer. The enhancer can be ligated to the 5, end or 3, end position of the coding sequence of the receptor binding agent in the (spHced int〇) vector, but is preferably located at the 51-terminal end of the promoter. For use in eukaryotic host cells (yeast, fungi, insects, plants, animals, humans, nucleated from other multicellular organisms) The expression vector of a nucleated cell may also contain sequences required for termination of transcription and for stabilization of mRNA. Such sequences are typically obtained from the untranslated region of the 5th and 'occasion 3' ends of eukaryotic or viral DNA or cDNA. The zone contains nucleotide segments which can be transcribed into polyadenylated fragments in the untranslated portion of the mRNA encoding the receptor binding agent. The performance vector can also include one or more intronic sequences. The receptor binding agent can also be expressed in insect cells, such as Sf9 or SF21 cells, for example, using the pFAST-BACTM system. Other exemplary baculovirus expression vectors are available from Invitrogen, Life.

Technologies, Inc., Carlsbad (Carlsbad, CA, USA). The receptor binding agent can also be expressed in mammalian cells. For example, cell lines derived from mammals can be used. Mammalian host cell

Examples of strains include COS-7 cell line of monkey kidney cells (ATCC CRL 1651) (Gluzman et al., Cell, 23: 175 '1981), L cells, C127 cells, 3T3 cells (ATCCCCL 163), Chinese hamster ovary (CHO) cells, human cervical cancer cells (HeLa cells)' and BHK (ATCC CRL)

68 201217528 10) Cell line 'and CV1/EBNA cell line derived from African green monkey kidney cell line CV1 (ATCC CCL 70), as described in the paper by McMahan et al. (EMBO Journal, 10: 2821, 1991). Established methods for introducing DNA into mammalian cells have been described in the literature. (Kaufman, R. J., "Large Scale Mammalian Cell Culture", 1990, p. 1569). Other methods suitable for the synthesis of receptor binding agents in recombinant cells are described in "Molecular Cloning: A Laboratory Manual", Third Edition, Sambrook et al. (ed.), Cold Spring Harbor Publishing, 2001 (ISBN number: 0879695773) ° Once expressed in cells, receptor binding agents can be recovered from cell culture fluids, inclusion bodies, or cell lysates. Cells can be broken by a variety of physical or chemical methods, such as: freeze-thaw cycling, sonicati〇n, mechanical disruption, or cytosol (ceu iysing) Agents) (eg detergents). The receptor binding agent can be purified from other cellular proteins or polypeptides that can be found in cell lysates or cell culture fluids. A variety of methods for protein purification can be employed and such methods are known and described in the art, for example, 'In Deutscher, Methods in Enzymology' 182, 1990; and Scopes, Protein Purification: Principles and Practice, 69 201217528

Springer-Verlag Publishing, New York, 2010 (ISBN number: 1441928332). Exemplary purification procedures include: fractionation on an ion-exchange column; ethanol precipitation; reverse phase HPLC; silica or cation exchange resin ( For example, DEAE) chromatography, chromatofocusing; polyacrylamide electrophoresis (SDS-PAGE); ammonium sulfate precipitation; gel filtration using, for example, Sephadex G-75 Protein A Sepharose columns are used to remove IgG-specific impurities, and affinity columns (eg, metal chelating columns) are used to bind epitope markers (epitope). A -tagged type of protein and a column having a plurality of ligands for binding to any purified moiety attached to the receptor binding agent. A purification process can include a combination of two different ion-exchange chromatography steps, such as cation exchange chromatography followed by anion exchange chromatography, or vice versa. The receptor binding agent can be eluded from the ion exchange resin by a variety of methods, including salt and/or pH gradients or step gradients & In some embodiments, the receptor binding agent comprises a purification moiety (e.g., an epitope tag, an affinity handle, and an affinity handle). These moieties can be used for affinity chromatography and optionally removed by proteolytic cleavage (piOteQlytie cleavage). A receptor binding agent can be stored in a variety of solvents, including: water, pBS,

70 201217528 with buffered solution. Exemplary buffer solutions include sodium acetate pH 4.5, sodium acetate pH 4.7, sodium acetate pH 4.9, sodium acetate pH 5.1, sodium acetate pH 5.3, sodium acetate pH 5.5, succinate pH 5.2, succinic acid Salt pH 5.4, tetanic acid salt pH 5.6, succinic acid salt pH 5.8, histidine pH 5.7, histidine pH 6.0, histidine pH 6.3, histidine 6.6, sodium sulphate Phosphate) pH 6.5, sodium phosphate pH 6.7, sodium phosphate 7.0, sodium phosphate pH 7.3' sodium phosphate pH 7.7, 11 m salidium (imidazole) pH 6.5, 11 m alpha sitting pH 6.8, 11 mw pH 7.2, trihydroxyl Tris, pH 7.0, Tris, pH 7.5, Tris, pH 7.7. The buffer can be, for example, about 1 to 100 millimolar (mM), 5 to 50 millimolar (mM) '10 to 50 millimolar (mM), or 5 to 25 millimolar (mM) ), the concentration exists. The solution may further comprise a salt such as sodium carbonate (NaC) (eg, 50 millimolar (mM), 150 millimolar (mM), or 250 millimolar (mM), Range), arginine (eg, at about 1%, 2%, 3%, 4%, 5%, 7.5%, in the range between them), sucrose (eg, at about 1) %, 2% ' 3%, 4% ' 5%, 7.5% ' 8.5% ' 10% ' 15%, the range between them), and / or glycerol (glycerol) (eg: at about 0 5 %, 1%, 2%, 33⁄4, 4%, 5% ' 7.5% ' 8.5% ' 10%, 15% ' and the range between them). The receptor binding agent may be present in an amount of at least 50 milligrams (mg), 100 milligrams (mg), 500 milligrams (mg) of 1 gram (g), 5 grams (g), 10 grams (g) or more. In the composition. Pharmaceutical Composition 71 201217528 A receptor binding agent can be formulated into a pharmaceutical composition. Typically the composition is sterile and has a buffer, a pharmaceutically acceptable salt, and one or more of an excipient or stabilizer. For example, the composition can be an aqueous composition. One of the receptor binding agents described in the present invention can be formulated according to standard methods for living organisms. See, for example, Gennaro (eds.), Remington: The Science and Practice of Pharmacy, 20th ed., Lippincott, Williams & Wilkins, 2000, (ISBN number: 0683306472) Ansel et al., "Pharmaceutical Dosage Forms and Drug Delivery Systems", 7th ed., published by Lippincott Williams & Wilkins Publishers, 1999 (ISBN number: 0683305727); Kibbe (ed.) Handbook of Pharmaceutical Excipients, 3rd edition, 2000, (ISBN number: 091733096X); "Protein formulation and delivery", McNally and Hastedt (eds.), Informa Health Care published, (ISBN number: 0849379490), 2007. A receptor binding agent for a pharmaceutical composition typically has at least 10'20' 50, 70, 80, 90' 95, 98' 99, or 99.99% purity, and typically does not contain human protein. It can be the only protein in the composition or the only active protein in the composition. It may also incorporate one or more other active proteins, such as one or more other purified active proteins, such as a related or unrelated protein. In some embodiments, the composition may comprise a concentration of the receptor binding agent in a boundary between about 〇〇 〇〇 〇 〇 , , , , ,

72 201217528 0·001 ~0·1%, 0·01 ～1%, or 0·1%~10% concentration. Accordingly, the present invention is also characterized by the purified and isolated forms of the agents described herein. The term "isolated" means that the material is removed from its original environment (e.g., the cell or material from which the receptor binding agent is produced). The pharmaceutical composition may be substantially free of pyrogenic materials, substantially free of nucleic acids, and/or substantially free of intracellular enzymes and components, such as: polymerases' ribosomal Proteins, with chaperone proteins - a pharmaceutical composition can include a pharmaceutically acceptable carrier. As the user of the present invention, "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal. Agents, isotonics and absorption agents, and physiologically compatible analogs. A "pharmaceutically acceptable salt" refers to a salt that retains the desired biological activity of the parent compound and does not cause any undesirable toxicological effects (see, for example, Berge, SM et al., J. Pharm. Sci., 66: 1-19, 1977, for example: acid addition salt and base addition salt. In one embodiment, the receptor binding agent is formulated with one or more excipients, such as: sodium chloride, and a phosphate buffer (eg, acid-filled hydrogen) Sodium dibasic phosphate heptahydrate, sodium sulphate 73 201217528 (sodium monobasic phosphate), and polysorbate. It can be, for example, provided with a concentration of 5 to 1 〇〇, 5 to 3 〇, 3 〇 to 5 〇, or 5 〇 to 1 〇〇 mg/ml (mg/ml), for example, a buffer In solution, and stored at a temperature of 2 to 8 ° C. The pharmaceutical composition can also be in a variety of other forms. Such forms include, by way of example, liquid, semi-solid, and dosage forms, such as liquid solutions (eg, injectable solutions and infusible solutions), Dispersion or suspensions ' with liposomes (lipos〇me). The preferred form may depend on the administration and the intended mode of the therapeutic application. The compositions described in this application for such agents are typically in the form of injectable or poorly soluble solutions, or for topical or ocular delivery (see below). Pharmaceutical compositions are typically sterile and stable under the conditions of manufacture and storage. A pharmaceutical composition can also be tested to ensure that it meets the regulatory and industrial standards (in (justry standard). The composition can be formulated as a one-bath microemulsion, dispersion, microlipid Liposome, or other 〇rdered structure suitable for high drug concentration. Sterile injectable solutions can be used in a suitable solvent to provide the required amount of one of the agents described in the present application. In combination with one of the above listed ingredients or the like, combined with the requirements 'and then prepared by filtered sterilization. Generally, the dispersion is as described in the present case. One of the agents is prepared by incorporation into a sterile vehicle containing a dispersion medium, and other components listed above are required. It is used to prepare a sterile powder of the sterilized liquid of the liquid 2012. The preferred preparation method is vacuum drying method (v_m dry ing) and cold; reezedrying, which produces the powder having the agent described in the present application plus any other desired The composition of the solution from the previous solution. The proper fluidity of the solution can, for example, utilize a coating such as lecithin, by maintaining the dispersion. The particle size is maintained by the use of a surfactant. The continuation of the injection composition can be designed by incorporating a delayed absorption agent such as monostearate salt and gelatin. For example, a receptor binding agent can be combined with a polymer, for example: a substantially non-antigenic polymer, such as a polymer A polyalkylene oxide or a polyethylene oxide. In some embodiments, the polymer is covalently attached to the receptor binding agent', eg, directly or indirectly. Suitable polymers will vary substantially in weight. Having a molecular number average weight ranges from about 200 to about 35,000 Daltons (or about 1, 〇〇〇 to About 15,00 A polymer system of 0, and from 2,000 to about 12,500 Daltons can be used. For example, a receptor binding agent can be conjugated to a water soluble polymer, for example: a hydrophilic poly A polyvinyl polymer, for example, a polyvinylalcohol or a polyvinylpyrrolidone. One non-limiting list of such polymers includes polyalkylene oxide homopolymers, such as polyethylene glycol (PEG) 75 201217528 or polypropylene glycol, poly. Polyoxyethylenated polyols, copolymers thereof and block copolymers thereof, are limited in terms of maintaining the water solubility of the block copolymer. Other useful polymers include polyoxyalkylenes, such as polyoxyethylene, polyoxypropylene, polyoxyethylene and polyoxypropylene (Pluronics); Acrylic acid (卩〇1>〇1161;11&£^>^165);carbomers; branched or unbranched polysaccharides comprising the following sugar monomers: D-mannose , D- and L-galactose, fucose, fructose, D-xylose, L-arabinose, D-gluconic acid ( D-glucuronic acid), sialic acid, D-galacturonic acid, D-mannuronic acid (eg polymannuronic acid) Acid), or alginic acid, D-glucosamine, D-galactosamine, D-glucose and neuraminic acid ), including homopolysaccharides and heteropolysaccharides like lactose, amylopectin, and starc h), hydroxy ethyl starch 'amylose, dextrane sulfate, dextran, dextrin, glycogen Or a polysaccharide subunit of acid mucopolysaccharides, such as: hyaluronic acid; a polymer of sugar alcohol, such as polysorbitol and polymannitol ( Polymannitol); heparin or heparan.

76 201217528 In certain embodiments, the receptor binding agent can be prepared with a carrier that will protect the compound from rapid release. It can be delivered in a controlled release formulation, either as an implant or a microencapsulated delivery system. Biodegradable, biocompatible polymers such as ethylene vinyl acetate, polyanhydride, polyglycolic acid, collagen can be used. (collagen), polyorthoester (polyorthoester), and polylactic acid (polylactic acid). Related background information can be found, for example, in "Sustained and Controlled Release Drug Delivery Systems", edited by J. R. Robinson, published by Marcel Dekker, Inc., New York, 1978. A drug-receptor binding agent can be administered to a subject, for example, a human subject, by a variety of different methods, such as by intravenous bolus or by continuous infusion over a period of time (continuous Infusion) in intravenous administration, intramuscular, intramuscular, intraarterial, intrathecal, intracapsular, intraorbital, heart Internal (intracardiac), intradermal, intraperitoneal, intrasynovial, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular ( Subcapsular), subarachnoid, intraspinal, epidural injection (epidural 77 201217528 injection), intrasternal injection and infusion. Other modes of administration include topical (eg, dermal or mucosal) or inhalation (eg, intranasal or intrapulmonary) routes. For many applications, the route of administration is one of the following: intravenous injection or infusion, subcutaneous injection, or intramuscular injection. The receptor can be a fixed dose. Or it can be administered at a dose of one milligram per kilogram (mg/kg). It can be administered intravenously (IV) or subcutaneously (SC). The receptor binding agent can be administered, for example, every day, every other day, every three, four or five days, every three weeks to three weeks, for example every four weeks, or every month. A pharmaceutical composition can include a "raperutically effective amount" of one of the agents described herein. One therapeutically effective amount may vary depending on various factors, such as disease state, age 'gender' individual weight, one of the compound elicit individuals. The ability to respond, such as at least one disease parameter improvement, or at least one symptom of the disease (and optionally the effect of any other agent being administered, a therapeutically effective amount is also a therapeutic A beneficial effect exceeds any toxic or detrimental effect of the composition. A receptor binding agent is typically administered in a therapeutically effective amount. The pharmaceutical composition may utilize a medical device, such as an implant, infusion Infusion pump, hypodermic needle, 78 201217528 and needleless hypodermic injection device. The device may include, for example, one or more for storing a pharmaceutical composition. Housings' and can be configured to deliver a unit dose of the receptor binding agent, optionally with one Two doses. The dose may be a fixed dose, that is, a physically discrete unit suitable for use as a unitary dosage for the individual to be treated; each unit may comprise a calculated The predetermined amount of the receptor binding agent of the desired therapeutic effect is combined with a pharmaceutical carrier and optionally with another agent. In some embodiments, to treat a disease as described herein, a receptor binding agent may To be administered to an individual having the disease in an amount and time sufficient to induce at least one indicator of a sustainedeci improve the severity of the disease. If the individual is between one and four An improvement in at least two of the weeks, the improvement can be considered "sustained." The degree of improvement can be determined based on the sign or symptom, and can also be provided A questionnaire is given to the individual, such as a quality-of-life questionnaire. A variety of indicators of the degree of illness can be assessed to determine the treatment. Whether the amount and time are sufficient. The baseline value of the selected indicator or indicator group is established by examining the individual prior to administration of the first dose of the receptor binding agent. Preferably, the baseline test is administered. The first dose is completed within about sixty days. The improvement is induced by re-administering a dose of the receptor binding agent, 79 201217528 until the individual exhibits an improvement over one of the selected indicators or indicator group baselines. . In the treatment of chronic diseases, the degree of improvement can be obtained by repeated administration for at least one month or more, for example, one, two or three months or longer, or indefinitely. In the treatment of an acute condition, the agent can be administered in a period of one to six weeks or even in a single dose. Although the extent of the condition can be shown to improve based on one or more indicators after treatment, the treatment can continue indefinitely at the same level or at a reduced dose or frequency. Treatment can also be interrupted, for example, depending on whether the symptoms improve or disappear. Once treatment is reduced or interrupted, treatment can resume if the symptoms recur. A therapeutic receptor binding agent, such as a receptor binding agent that binds to IL-1RI and antagonizes IL-1 signaling, can be used to treat an "IL-1 mediated disorder" , including any disease or medical condition, is (i) at least partially caused by il-1 agonism, (ii) with an elevated level or activity of IL- 1 signalling component (eg IL-Ια, IL-Ιβ, or IL-1RI) or elevated IL-1 signal transmission, and/or (iii) by reducing IL-1 activity Ameliorated. Diseases mediated by IL-1 include acute and chronic diseases, including autoimmune diseases and inflammatory diseases. Diseases mediated by IL-1 include systemic and non-systemic (non -systemic) disease. It has been recognized that IL-la and IL-Ιβ are effective triggers for infectious and inflammatory diseases including rheumatoid arthritis.

80 201217528 Pro-inflammatory cytokines. Increased IL-1 production is observed in patients with multiple autoimmune diseases Outoimmune disorders, ischemia, and several cancers, suggesting that IL-1 is implicated in these and related diseases. . For related background information, see Sims and Smith, IL-1 family: regulators of immunity, Nature Reviews Immunology, doi:10.1038/nri2691, 2010 . The term "treat" means to effectively improve a condition, symptom, or a parameter associated with a disease (such as one of the diseases described herein), or to avoid a disease progression (progression) ) to a statistically significant extent or to a degree that is familiar to the artist, one of the 'manner' and/or mode, for a body, such as a patient' The administration of one of the agents. The treatment can be used for cure, heai, ausuation, reIieve, alter, remedy, ameliorate, palliate, The disease is improved or affected, the symptoms of the disease, or the predisposition towards the condition. An effective TT' mode, or mode may vary depending on the individual or may be tailor made for the individual. Exemplary individuals include humans, primates, and other non-human suckling animals. A receptor binding agent can also be administered prophylactically to reduce the risk of a disease or its symptoms. The disease intervened by IL_1 can be an autoimmune disease. Examples of autoimmune diseases intervened by IL-1 include: rheumatoid arthritis (rheum.at〇id 81 201217528 arthritis), ankylosing spondylitis, Behcet's syndrome, inflammation Inflammatory bowel disease (including Crohn's disease and ulcerative colitis) 'asthma, psoriasis, type I diabetes' and others The disease indicated in this case. One of the receptor binding agents described in the present invention can be administered to an individual at or at risk of an autoimmune disease mediated by such IL-1. Illustrative ratios include: The diseases that are intervened include: rheumatoid arthritis and related arthritides. A receptor binding agent can be used to treat rheumatoid arthritis or One of the risks is rheumatoid arthritis (RA), a chronic systemic autoimmune inflammatory disease that affects the joint synovial membrane and damages articular cartilage (articuiar cartilage) The pathogenesis of rheumatoid arthritis (RA) is T lymphocyte-dependent and may include the production of autoantibodies known as rheumatoid factors. A complex of rheumatoid factor and antigen can form and accumulate in joint fluid and blood, inducing lymphocytes, neutrophils, and infiltration into monocyte synovium (synovium) The joint system is usually affected by a symmetrical pattern, but extra-articular disease can also be developed. Health, for example, causes pulmonary fibrosis, vasculitis, and cutaneous ulcers, or manifests as a neutrophil

82 201217528 Reduces (neutropenia) 'thrombocytopenia and splenomegaly's Ferti's syndrome (Feity, s syndr〇me). The patient may also be in the affected joint 'pericarditis, pleuritis 'coronary arteritis, and the area of interstitial pneumonitis with pulmonary fibrosis. Rheumatoid nodules (rheumat〇id nodules) are presented. One form of IL-IRa is used to indicate moderate to severe active rheumatoid arthritis (RA). See, for example, Cohen, S. et al., Arthritis & Rheumatism, 46, 614-24, 2002; Fleischmann, RM et al., Arthritis & Rheumatism ), 48, 927-34, 2003; Nuki, G. et al., Arthritis & Rheumatism, 46 ' 2838-46 ' 2002; Schiff, MH et al., Arthritis and rheumatism Arthritis & Rheumatism, 50, 1752-60, 2004. Symptoms of active rheumatoid arthritis (active RA) include fatigue (lati of appetite), low grade fever, muscle and joint aches, and stiffness. . Muscle and joint stiffness are usually most pronounced after the morning and inactivity. During the flare period, joints often become reddened, swollen, painful, and tender, generally due to synovitis. The scale used to assess the symptoms of rheumatoid arthritis (ra) and rheumatoid arthritis (RA) includes the Rheumatoid Arthritis Severity Scale (RASS; Bardwell et al., 2002, Rheumatology). Rheumatology, 83 201217528 41(1): 38-45), SF-36 Arthritis Specific Health Index (ASHI; Ware et al., 1999, Journal of Medical Care (Med) . Care.), 37 (5 Suppl): MS40-50), Arthritis Impact Measurement Scales or Arthritis Impact Measurement Scales 2 (AIMS or AIMS2; Meenan et al. Human, 1992, Arthritis Rheum., 35(1): 1-10); Stanford Health Assessment Questionnaire (HAQ), HAQII, or revised HAQ ( See, for example, Pincus et al., 1983, Arthritis Rheum, 26(11): 1346-53). A receptor binding agent described in this case can be administered to an individual at or at risk of rheumatoid arthritis (RA) to delay its onset and/or ameliorate - or a plurality of the foregoing Signs and symptoms (symptom). The individual may have moderate to severe active rheumatoid arthritis (active RA). The individual can be one of TNF inhibitor therapy (eg, ENBREL® (etanercept), remedy® (adalimum) (HUMIRA® (adalimumab) )), or a non-responder under the treatment of Remi-Cade® (infliximab); the individual may have been previously administered a TNF inhibitor; or the individual It is also sustainable to receive a TNF inhibitor (and react or not react). The individual can also be administered methotrexate. The individual can be administered one or more other DMALDS (disease modified antirheumatic drugs)

84 201217528 (disease modifying anti-rheumatic drugs)), a corticosteroid, and/or a non-steroidal anti-inflammatory. Other drugs that can be co-administered with a receptor binding agent include inhibitors of CD28 (eg CTLA4-Ig), inhibitors of RANKL, scare 'IL-6, IL-8, and IL-17 . Inhibitors include antibodies to such agents, specific to the soluble receptors of these mediators' and/or antibodies to the receptors of such mediators. Juvenile chronic arthritis. A receptor binding agent can be used to treat juvenile chronic arthritis', e.g., in an individual below 21, 18, 17, 16, 15, or 14 years of age. Juvenile chronic arthritis is similar to rheumatoid arthritis (ra) in several respects. The individual may be a rheumatoid factor positive person. The individual may have a Pauciarticular, polyarticular' or systemic form of the disease. This arthritis can cause joint ankylosis and retarded growth and can also lead to chronic anterior uveitis and systemic amyloidosis. A receptor binding agent can be used to delay the onset or ameliorate of one or more of the symptoms - or a plurality of such symptoms. A receptor binding agent can be used to treat juvenile idiopathic arthritis, including systemic onset juvenile idiopathic arthritis (SO-JIA). The individual may have failed corticosteroid treatment (corticosteroid 85 201217528 treatment) or need to be treated with corticosteroids at a dose equivalent to or exceeding 0.3 mg/kg (mg/kg) per day. See, for example, Quartier et al., 2011, Ann Rheum Dis., 70(5): 747-54. Other rheumatic diseases (Rheumatic D丨sorders). One of the receptor binding agents described in this case can also be used to treat other rheumatic diseases, including scleroderma, systemic lupus erythematosus (syStemic lupus erythematosus), gout, osteoarthritis. , rheumatoid polymyalgia (polymyalgia rheumatica), psoriatic arthritis and chronic Lyme arthritis, voluntary muscle and other muscle inflammation, including the skin muscle Dermatomyositis, inclusion body myositis, polymyositis, lymphangioleimyomatosis, pyrogenic arthritis syndrome, pediatric granulomatous joints Pediatric granulomatous arthritis (PGA) / Blau syndrome, and other rheumatic diseases discussed in this case. A receptor binding agent can be used to treat metabolic rheumatic diseases, such as: diseases associated with hyperuricemia, such as gout, including chronic acute gout, and other crystallizations. Crystal-mediated arthropathies. The agent can be used to treat medication-induced gout-related flies, including, for example, xanthine oxidase inhibitors, urate oxidase, or uric acid. A disease induced by uricosuric agents. In the case of gout, uric acid crystals can activate hair

86 201217528 Inflammasome and triggers the release of IL-Ιβ. Spondyloarthropathies. A receptor binding agent can be used to treat spondyloarthropathies, including diseases such as ankylosing spondylitis, Reiter's syndrome, associated with inflammatory bowel disease. Arthritis associated with inflammatory bowel disease 'spondylitis associated with psoriasis, juvenile onset spondyloarthropathy and undifferentiated spondyloarthropathy (undifferentiated) Spondyloarthropathy). Spinal joint disease (spondyloarthropathies) is often associated with the HLA-B27 gene. An individual may be deficient in rheumatoid factor and may exhibit sacr〇iiiitis with or without spondylitis and inflammatory asymmetric arthritis. Individuals may also have ocular inflammation (see below). Scleroderma. A receptor binding agent can be used to treat scleroderma or SyStemic sclerosis. Scleroderma is characterized by a localized or systemic skin induration. It can also occur in vascular lesions and endothelial ceU injury at the microvascular (mjcrovascuiature). Individuals may present mononuclear cells infiltrating cutaneous lesions and have anti-nuclear antibodies. Other organs that exhibit path 〇 genesis may include: gastrointestinal tract, which may have smooth muscle (smo〇th 87 201217528 muscle) atrophy and cause peristalsis/motility (motility) Fibrosis; kidney, which may have concentric subendothelial intimal proliferation, resulting in reduced renal cortical blood flow in arcuate and interlobular arteries (renal cortical blood flow), and which can lead to proteinuria (pr〇teinuria), azotemia and hypertension; skeletal muscle, which may involve atrophy, interstitial fibrosis (interstitial) Fibrosis), inflammation, lung, interstitial pneumonitis and interstitial fibrosis; and the heart, which can present, for example, contraction band necrosis Scarring / fibrosis. One of the receptor binding agents described herein can be administered to an individual suffering from or at risk of scleroderma to ameliorate one or more of the aforementioned signs and symptoms. Sjogren’s syndrome. A receptor binding agent can be used to treat Sjogren's syndrome. Sjogren's syndrome is characterized by immune-mediated inflammation and subsequent functional destruction of the tear glands and salivary glands. The disease can be associated with or associated with inflammatory connective tissue diseases. The disease is associated with the production of autoantibodies against R〇 and La antigens, both of which are small RNA-protein complexes. Lesion 88 201217528 (Lesion) can cause keratoconjunctivitis sicca, xerostomia, along with other biliary cirrhosis, peripheral neuropathy or sensory neuropathy, and The manifestations or associations of palpable purpura. The treatment of eye diseases associated with Sj6gren's disease is also discussed below. Thyroid Disorders. A receptor binding agent can be used to treat thyroid disorders. Illustrated thyroid disorders include Graves, disease, Hashimoto's thyroiditis, juvenile lymphocytic thyroiditis, and atrophic squamous glands Atrophic thyroiditis' is the result of an autoimmune response against thyroid antigen, which produces antibodies that react with proteins present on the thyroid gland and usually specific to the sacral gland. Available experimental models include spontaneous models: rats (BUF and BB rats) and chickens (obese chicken strain). And inducible models for animals by thyroglobulin, or thyroid microsomal antigen (thyroid peroxidase) Established by immunization. Diabetic and Metabolic disorders. A receptor binding agent can be used to treat diabetic disorders, such as juvenile onset diabetes (including autoimmune diabetes mellitus and insulin-dependent diabetes (insulin- Dependent types of diabetes) and maternal onset diabetes (including non-insulin dependent and 〇besity_mediated diabetes), type I diabetes and type 2 Type II diabetes. For example, 'type 1 diabetes (typeidiabetes) or insulin-dependent diabetes is caused by autologous antibodies and auto-reactive T cells. (pancreatic islet cell) is associated with autoimmune destruction. In addition, reducing the activity of iL-ΐβ promotes glucose control and beta cell function and can be used to treat type II diabetes. See, for example, 〇wyang et al., Endocrinology, 2010; 151(6): 2515-27. For example, in some embodiments, one of the receptor binding agents described herein can be administered to an individual who is not currently being administered insulin, e.g., the individual is not an insulin dependent. For example, the individual can be a pre-diabetic. The individual may have impaired glucose tolerance or impaired fasting glucose. The individual can be obese or have a body mass index above 23, 25, 30, 35, or 40 (kg/m2 (kg/m2)). The individual can be insulin resistant, and/or have the characteristics of hyperglycemia or hyperinsulinemia. The individual may be at risk of having a progression to type II diabetes. See also 90 201217528

Larsen et al., 2007, New England Journal of Medicine (NEJM), 356: 1517-26 and Larsen et al., 2009, Diabetes Care, 32: 1663-8. In some embodiments, the individual has a fasdng plasma glucose greater than 6"' 6.5, 7, or 8 millimoles # (mmol/L). In some embodiments, the individual has an A1C level (A1C level) above 55, 5.7', 6, 6.4, 7, 7.5, or 8%. In some embodiments, the individual can also be administered a secretagogue, such as, for example, a sulfonylurea that stimulates insulin secretion (eg, chlorpropamide, durazolamide) (tolazamide), acetohexamide, tolbutamide, gyiburide, glimepiride 'glipizide' and/or meige Megiitinides (eg repaglinide, nategiinide). The individual can also be administered a pair of biguanides (e.g., metformin). The receptor binding agent can be administered to reduce the loss of pancreatic beta cells and/or damage thereto. In some embodiments, the individual is located at rs4251961 near the 5' end of the gene IL1RN. The C allele is an allogeneic combination of heterozygous or homozygous. Therapeutic effects of a receptor binding agent include ameliorating or avoiding the deterioration of secondary conditions associated with diabetes, such as diabetic retinopathy, renal transplant rejection in diabetic patients. , obesity-mediated insulin resistance, and renal failure 91 201217528 (renal failure), which may itself be associated with proteinuria and hypertension (Gastrointestina 丨 Disorders). The receptor binding agents described in the present invention can be used to treat inflammatory gastrointestinal disorders, including, for example, coeliac disease, Crohn's disease, ulcerative colitis. ), idiopathic gastroparesis pancreatitis, including chronic pancreatitis, acute pancreatitis, inflammatory bowel disease and ulcers, These include gastric and duodenal ulcers. Pulmonary disorders. A receptor binding agent can be used to treat lung diseases mediated by IL-1. Exemplary lung diseases that can be treated include chronic obstructive pulmonary disease (eg, emphysema and chronic bronchitis), alveolar proteinosis (pUim〇nary alveolar proteinosis) ), bleomycin-induced pneumopathy and fibrosis, pulmonary fibrosis, including idiopathic pulmonary fibrosis and radiation-induced pulmonary fibrosis (radiation- Induced pulmonary fibrosis), pulmonary sarcoidosis, cystic fibrosis, accumulation of collagen in the lung (c〇iiagen accumulation), adult respiratory distress syndrome (ARDS), pulmonary bronchial hypoplasia (broncho-pulmonary dysplasia) (BPD)), chronic obstructive pulmonary disease

S 92 201217528 Chronic obstructive pulmonary diseases, and chronic fibrotic lung disease of preterm infants. In addition, the receptor binding agents described in this case can be used to treat occupational lung diseases, including asbestosis 'coal worker's pneumoconiosis, silicosis or A similar condition associated with prolonged exposure to fine particles. Inflammatory and fibrotic lung diseases include eosin〇phiiic pneurn〇nia, idiopathic pulmonary fibrosis, and allergic pneumonia that may involve a misregulated immune inflammatory response (hypersensitivity pneumonitis), which can be treated with a receptor binding agent. Sarcoidosis is a disease of unknown etiology. It is characterized by the presence of epithelioid granulomas in almost any tissue of the body; this condition is most common in the lungs. Its pathogenesis is involved in the site of the disease, the activation of activated macrophages and the persistence of lymphoid cells' and subsequent local and systemic activity of the active products released by these cell types. The chronic sequelae caused by the release of the disease (Cardiovascular Disorders). One of the receptor binding agents described in the present invention can be used to treat cardi〇 vascular disorders or lesions such as aortic aneurysms, acute coronary syndromes, arteritis. 93 201217528 (arteritis), vascular occlusion, including cerebral artery occlusion, complications of coronary by-pass surgery, ischemia/reperfusion injury (ischemia/ Reperfusion injury), heart disease, including atherosclerotic heart disease, myocarditis, including chronic autoimmune myocarditis and viral myocarditis, heart Heart failure, including chronic heart failure, congestive heart failure, myocardial infarction, after heart surgery or ballooning of carotid artery Vasodilation surgery (balloon angioplastic pro After cedure), restenosis and/or atherosclerosis, silent myocardial ischemia, left ventricular ump function, left ventricle (left ventricle) Left ventricular) post implantation complications, Raynaud's phenomena, thrombophlebitis, including the vasculature of Kawasaki's vasculitis Vasculitis, veno-occlusive disease, giant cell arteritis, Wegener's granulomatosis, and Xuan-Henoch's purpura (Schoenlein-Henoch) Purpura). The receptor binding agent can also be provided prophylactically, for example, to reduce the risk of such cardiovascular disorders. In some embodiments, the receptor 94 201217528 binding agent is administered to a patient to treat or reduce the risk of atherosclerosis. The signal transduction of IL-1 mediation is activated by acute myocardial infarction and can induce apoptosis (ap〇pt〇tic cdl death) of peri-infarct myocardial cells. The size of the infarct area. One of the receptor binding agents described in the present invention can be administered to reduce the damage caused by myocardial infarction. For example, the receptor binding agent can be administered to an individual at risk of myocardial infarction, or to an individual who has experienced myocardial infarction, particularly, for example, at the most recent 2, 4, 6, 12, 24, or Within 48 hours, an individual undergoing an acute myocardial infarction. The agent can be administered in combination with other agents including, for example, heparin and aspirin. A receptor binding agent can also be used to treat stroke, sub-arachnoid hemorrhage, and/or head trauma or brain injury. For example, elevated concentrations of il·1 beta are involved in neuroinflammation associated with stroke and brain injury (Rothwell, NJ, et al., Journal of Neuroscience Trends (TINS) 23 (12 ) : 618-625, 2000). The receptor binding agent can be administered to reduce such inflammation and other inflammation associated with ischemia and/or hypoxia. In addition, the receptor binding agent can also be provided prophylactically, for example, to reduce the risk of such diseases and/or inflammation associated with such diseases. For example, the receptor binding agent can be administered to a risk of a stroke 'ischemic event, or a hemorrhagic event (such as sub-arachnoid hemorrhage). An individual of 201217528 may be given a drug, such as an individual who has experienced a stroke, ischemic condition, or bleeding (such as subarachnoid hemorrhage) within the last 2, 4, 6, 12, 24, or 48 hours. . Urinary tract disease and renal dysfunction (Genitourinary and Renal Disorders). The disease of the genitourinary system can also be treated with one of the receptor binding agents described in the present invention. These diseases include glomerulonephritis, including autoimmune glomerulonephritis, glomerulonephritis due to exposure to toxins or haemolytic streptococci Or other infectious agents infection secondary secondary glomerulonephritis (glomerulonephritis). Renal diseases of immune mediation, including glomerulonephritis and tubulointerstitial nephritis, which are the damage of renal tissue by antibodies or T lymphocytes. Directly producing autoreactive antibodies or T cells against the results of renal antigens, or indirectly depositing antibodies and/or immune complexes in the kidney, reaction Sexually against the results of other non-renal antigens. Therefore, other immune-mediated diseases that cause the formation of immune-complexes can also induce immune-mediated kidney disease as an indirect sequelae. Both direct and indirect immune mechanisms cause an inflammatory response that produces/induces the development of lesions in the renal tissue, with the results of organ damage (impairment) of 96 201217528 and In some cases it worsens to renal failure. Both the humoral immune mechanism and the cellular immune mechanism are involved in the pathogenesis of the disease. Ο

A receptor binding agent can also be used to treat uremk syndrome and its clinical complications (for example, renal failure, anemia, and hypertrophic). Cardiomyopathy)), including uremic syndrome associated with exposure to environmental toxins, drugs or other causes. The complications resulting from the absorption of the absorptive function due to inflammation of the gallbladder wall can also be treated. These complications include cholelithiasis (gallstones) and choliedocholithiasis (bile duct stones) and recurrence of cholelithiasis and choliedocholithiasis (recurrence). Further treatable diseases are complications of hemodialysis; prostate conditions 'including benign prostatic hypertrophy' nonbacterial prostatitis and chronic prostatitis ; and complications of hemodialysis. A receptor binding agent can also be used to treat chronic pain conditions, such as chronic pelvic pain, including chronic prostatitis/pelvic pain syndrome. With post-herpetic pain. 97 201217528 Hematologic and Oncologic Disorders. One of the receptor binding agents described in this case can be used to treat a variety of different forms of cancer, including acute myel〇genous leukemia, chronic myelogenous leukemia, EBV-positive nasopharyngeal carcinoma (Epstein-Barr virus-positive nasopharyngeal carcinoma) 'glioma, colon, stomach, prostate, renal cell, cervical (cervical) and ovary (ovarian) Cancer, lung cancer (SCLC and NSCLC), including cancer-related cachexia 'fatigue', asthenia's cachexia's paraneoplastic syndrome With hypercalcemia. See, for example, Voronov et al., 2003, Proceedings of the National Academy of Sciences (PNAS) 100: 2645-2650 (Voronov et al_ (2003) PNAS 100: 2645-2650). Solid tumors, including sarcoma, osteosarcoma, and cancer, such as adenocarcinoma (for example, breast cancer) and squamous epithelial cells Squamous cell carcinoma is also treatable. For the role of IL-Ιβ in certain tumors, see, for example, Krelin et al., 2007, Cancer Res > 67 · 1062-1071 (Krelin et al. (2007) Cancer Res. 67:1062-1071). Other cancers include esophogeal cancer, gastric cancer, gall bladder carcinoma, leukemia, including acute myelogenous leukemia, chronic myelogenous s 98 201217528 Leukemia), myeloid leukemia, chronic or acute lymphoblastic leukemia and hairy cell leukemia. Other malignancies with invasive metastatic potential, including multiple myeloma, can be treated with such receptor binding agents. See, for example, Lust et al., Maya, Mayo Clin Proc, 84(2): 114-122.

A receptor binding agent can also be used to treat anemias and hematologic disorders, including chronic idiopathic neutropenia, chronic anemia, regenerative anemia (aplastic anemia), including Fanconi's aplastic anemia; idiopathic thrombocytopenic purpura (ITP); thrombotic thrombocytopenic purpura, myelodysplastic syndrome (myelodysplastic syndromes) (including refractory anemia), refractory anemia having a refractory anemia with a ringed sideroblast, with excess blast Refractory anemia - refractory anemia with excessive transformation; myelofibrosis / myeloid metaplasia; and sickle cell vascular Sickle cell vasocclusive crisis. Autoimmune hemolytic anemia, immunity pancytopenia, and paroxysmal 99 201217528 paroxysmal nocturnal hemoglobinuria can be caused by and expressed in red blood cells (and In some cases, other blood cells, including platelets, are produced by the production of antibodies that react with antigens on the surface, and are mediated and/or ADCC/Fc-receptor-mediated by complement. The mechanisms (ADCC/Fc-receptor-mediated mechanisms) are used to remove the results of those cells coated with antibodies. Autoimmune thrombocytopenia (including thrombocytopenic purpura), and immuno-mediated thrombocytopenia in other clinical settings, antibody or patch Attachment to platelets and subsequent platelet destruction/removal results by complement lysis, ADCC or FC-receptor mediated mechanisms. The receptor binding agent can also be administered to an individual suffering from or at risk of a plurality of lymphoproliferative disorders, including autoimmune lymphoproliferative syndrome (ALPS). ), chronic lymphoblastic leukemia, hairy cell leukemia, chronic lymphatic leukemia, peripheral T-cell lymphoma, small lymphoid lymphoid Small lymphocytic lymphoma, mantle cell lymphoma, follicular lymphoma, Burkitt's lymphoma, EBV-positive T-cell lymphoma (Epstein-Barr) Virus-positive T cell lymphoma), histiocytic lymphoma, Hodgkin's disease, diffuse aggressive lymphoma 5 acute lymphatic leukemias, Τγ T gamma lymphoproliferative disease, cutaneous B cell lymphoma, cutaneous T cell lymphoma (ie, mycosis fungoides) and cezuri Sezary syndrome ° Hepatic Disorders. The receptor binding agents disclosed in the present disclosure are also useful for treating liver disorders such as hepatitis, including acute alcoholic hepatitis, acute drug-induced or viral hepatitis. , hepatitis A, B and C 'sclerosing cholangitis, hepatic sinusoid epithelium, and inflammation of the liver due to unknown causes. Hearing Disorders. Receptor binding agents can also be used to treat diseases involving hearing loss as well as diseases associated with abnormal IL-1 expression. Such diseases include cochlear nerve-associated hearing loss, which is thought to originate from an autoimmune process, such as autoimmune hearing loss, Meniere's Meniere's syndrome and cholesteatoma, a type of often associated with hearing loss, 101 201217528 middle ear disorder. Bone Disorders. Non-arthritic disorders of bones and joints can also be treated with the receptor binding agents described herein. This encompasses osteoclast disorders that cause bone loss, such as, but not limited to, osteoporosis (including osteoporosis (p0st-menopausal osteoporosis)), bone and joint Osteoarthritis, periodontal disease (toiodontitis) that causes tooth loosening or loss, and prosthesis loosening after joint replacement (usually with wear) Related to the inflammatory response of wear debris, for example: orthopedic implant osteolysis (Amy丨oid Disorders). Further, the receptor binding agent described in the present invention can be It is used to treat primary amyloidosis and secondary amyloidosis, which are characteristic of many different conditions, including Alzheimer's disease. Secondary reactive amyloidosis; Down's syndrome; and dialysis phase Dialysis-associated amyloidosis. In addition, receptor binding agents can be used to treat Amyotrophic lateral sclerosis (ALS) 'f· > Huntington's disease, and Parkinson's disease . These diseases also involve the formation of aggregates and ainyl〇ids that trigger inflammatory reactions.

S 102 201217528 Neurological Disorders. Receptor binding agents can also be used to treat demyelinating diseases in the central and peripheral nervous systems, including multiple sclerosis; spontaneous demyelinating polyneuropathy ( Idiopathic demyelinating polyneuropathy or Guillain-Barre syndrome; and chronic inflammatory demyelinating polyneuropathy. These diseases have a basis for autoimmunity and lead to nerve demyelination as a result of direct damage to oligodendrocytes or myelin. It is induced in multiple sclerosis disease and involves T lymphocytes. Multiple sclerosis has a relapsing-remitting course or a chronic progressive course. The lesion contains infiltration of microglial cells and infiltrating macrophages, which are mainly mediated by the τ lymphocytes; the CD4+T lymphocytic line is the main cell type at the lesion. The mechanism of cell death and subsequent demyelination of oligodendrocytes remains unclear but may be driven by τ lymphocytes. Myopathies. Receptor binding agents can be used to treat myopathies associated with inflammation and autoimmunity. Idiopathic inflammatory myopathies include dermatomyositis, polymyositis and other chronic muscle inflammation that causes muscle weakness due to unknown causes. 103 201217528 (chronic muscle inflammation) disease. Injury/inflammation is usually systemic and progressive. Autoantibodies (aut〇antibodies) are associated with most types. These myositis-specific autoantibody systems target and inhibit components involved in protein synthesis, proteins and RNA. Vasculitis Disorders. One of the receptor binding agents described in the present invention can be used to treat a vasculitis disorder, such as a systemic vasculitis. In the case of systemic vasculitis, the primary lesion is an inflammatory response, and subsequent damage to the blood vessel causes a deficiency in the tissue supplied by the damaged vessel. Blood/necrosis/degeneration, and in some cases the final end-organ dysfunction. Vasculitides can also occur as secondary lesions of other immune-mediated mediated diseases such as rheumatoid arthritis, systemic sclerosis, or the like. The sequelae, especially in diseases that are also associated with the formation of immune complexes. Diseases in the primary systemic vasculitis group include systemic necrotizing vasculitis, polyarteritis nodosa, allergic vasculitis (allergic) Angiitis) and granulomatosis, polyangiitis; Wegener's

S 104 201217528 granulomatosis); lymphomatoid granulomatosis; and giant cell arteritis. Miscellaneous vasculitides include: mucocutaneous lymph node syndrome (MLNS or Kawasaki's disease), isolated CNS vasculitis 'Bese Behcet's disease, thromboangiitis obliterans (Buerger's disease), and cutaneous necrotizing venulitis. It is believed that the pathogenic mechanism of these vasculitis disorders is mainly due to the deposition of immunoglobulin complexes in the vessel wall and subsequent inflammatory responses via ADCC, complement activation or both. CAPS (CIAS1-related periodic syndrome) (CAPS). A receptor binding agent can be used to treat a CAPS disease, i.e., CIAS1 Associated Periodic Syndrome (CIAS1 Associated Periodic Syn syndrome). CAPS includes three genetic syndromes: the Neonatal Onset Multisystem Inflammatory Disorder (NOMID) and the Muckle-Wells Syndrome (MWS). Familial Cold Autoinflammatory Syndrome (FCAS) with familial cold. (Hoffman et al., 2001, Naure, 29: 301-305; Feldmann et al., 2002, Am J Hum Genet, 71: 198-203; Aksentijevich et al., 2002 Year, Arthritis Rheum 46 : 3340-3348). CAPS can be inherited in an autosomal dominant manner by a sporadic or faniiiial pattern of 105 201217528. CIAS1 encodes a protein component of the NALP3 'inflammasome', a cytosine caspase 1 active subcellular enzyme complex. Mutations on CIAS1 result in increased IL-1 production and many pathological outcomes (Aksentijevich et al., 2002, supra). IL-1 strongly induces the production of acute phase reactants in the liver, such as C-reactive protein (CRP) and serum amyloid A (SAA). CAPS disease has common clinical features and presents a spectral range of clinical severity. NOMID is the most severely disabling, M WS is slightly less severe, and FCAS is the least severe. CAPS disease has several overlapping features and can each have unique signs and constellations. Common to all of these diseases are fever, urticaria-like rash, arthritis or arthralgia, myalgia, malaise, and conjunctivitis. In NOMID, chronic aseptic meningitis may cause mental retardation and these patients may cause disfiguring and obstruction of bone at the epiphyses and patilae. Disabling bony overgrowth. These patients may also become s 106 201217528 suffer blindness due to an increase in intracranial pressure leading to optic nerve atrophy. MWS and NOMID systems are often associated with severe inflammatory responses' which may include auditory systems, meninges, and joints. These patients may have high spiking fevers per spike and a chronic rash that often changes distribution and intensity. The patient may have hearing loss or deafness. Conjunctivitis and papilledema are often observed. Amyloidosis may develop and cause renal failure (renai faiiure) due to chronic inflammatory reactions and excessive production of acute phase reactants (especially SM). An acceptor binding agent can be administered to an individual having a NOMID, MWS, or FCAS or an individual diagnosed with a genotype associated with NOMID, MWS, or FCAS. In addition, a receptor binding agent can be administered to an individual having TRA receptor (TNF receptor associated periodic syndrome). Dermatological disorders. A receptor binding agent can be used to treat dermatological disorders, such as psoriasis, or other autoimmune or immune-mediated skin diseases. Autoimmune or immune skin diseases include bullous skin diseases, erythema multiforme, and contact dermatitis, which are mediated by autoantibodies. The occurrence is T lymphocyte-dependent. Psoriasis is a lymphocyte-mediated inflammatory disease. The lesion contains T lymphocytes 107 201217528 spheres, infiltration of macrophages with antigen processing cells and some neutrophils. Other treatable skin or mucous membranes include acantholytic diseases, including Darier's disease, keratosis follicularis and pemphigus. Vulgaris). Further diseases include: acne, acne rosacea 'alopecia areata', aphthous stomatitis, bullous pemphigoid, burns, Eczema, erythema, including erythema multiforme and erythema multiforme bullosum (Stevens-Johnson syndrome), inflammatory Inflammatory skin disease, lichen planus, linear IgA bullous disease (chronic bullous dermatosis), loss of skin elasticity ( Loss of skin elasticity, including mucosal surface ulcers, including gastric ulcers, neutrophilic dermatitis (Sweet's syndrome) dermatomyositis, hair Pityriasis rubra pilaris, psoriasis, pyoderma gangrenosum, multicentric reticular group Cell histiocytosis (multicentric reticulohistiocytosis), and toxic epidermal dissolved (toxic epidermal necrolysis). Other skin-related conditions that can be treated with receptor binding agents include dermatitis herpetiformis (Du Lin)

108 201217528 Duhring's disease, atopic dermatitis, contact dermatitis, and urticaria (including chronic idiopathic urticaria) . Allergic Disorders. A receptor binding agent can be used to treat allergic disorders such as asthma (asthma); allergic rhinitis; atopic dermatitis; food allergy (f〇〇d hypersensitivity) With urticaria. These diseases are usually mediated by T lymphocyte-induced inflammatory response, IgE-mediated inflammatory-inflammation or a combination of the two. Asthma is a chronic condition involving the respiratory system. The airway occasionally constricts, sends k (becomes inflamed) ' and has excessive excesses of mucus, usually caused by one or more triggers. May be exposed to an environmental stimulant (or allergen), such as cold air, warm air 'fishing air' exercise or exercise "emotional stress" and viral disease (virai illness) triggers the episode (Episodes). Airway narrowing can cause symptoms like wheezing, shortness of breath, chest tightness, and coughing (C0Ughing). A receptor binding agent can be used to treat asthma and can be formulated for local or pulmonary delivery for treatment or can be delivered parenterally. Transplantation. A receptor binding agent can be used to administer to 109 201217528 an individual undergoing transplantation 'undergoing transplant' or recovering from a transplant. Transplantation associated diseases, including graft rejection (graft rejects) and graft-versus-host-disease (gvhd), T lymphocyte-dependent; inhibition of T lymphocytes The function can be improved (ameliorative). Corneal transplantation can be associated with neovascularization, which can be improved by treatment with a receptor binding agent. A receptor binding agent can also be used to treat solid organ transplantation (such as the heart, liver 'skin' kidney, lung (lung transplant airway obliteration)) or other transplants (transplants) (including complications caused by bone marrow transplants). Infectious Diseases. The receptor binding agents described in this case are useful for treating protozoal diseases, including malaria and schistosomiasis and erythema nodosum leprosum; bacterial or viral Bacterial or viral meningitis; tuberculosis, including pulmonary tuberculosis; and bacterial or viral infection (including influenza infection and infectious mononuclear blood cells) Infectious mononucleosis) Pneumonia (pneumonitis)

Inherited periodic fever syndrome can also be treated with a receptor binding agent, including familial Mediterranean fever, high immunoglobulin D

S 110 201217528 (hyperimmunoglobulin D) and periodic fever syndrome and TNF-receptor associated periodic syndromes (TRAPS), and Adult-onset Still's Disease), Schnitzler's syndrome, and fibrosing alveolitis. In some embodiments, a receptor binding agent is administered to a subject to reduce the activity or performance of IL-6 in the individual. For example, the individual can have a disease associated with IL-6 or at least partially mediated by IL-6. Ocular Disorders and Ocu丨ar Delivery The receptor binding agents described in this case can be used to treat ocular diseases, including eye diseases affecting the surface of the eye, inflammatory eye diseases (inflammatory Ocular disorders, with ocular disorders that are at least partially mediated by an autoimmune response. In some embodiments, the ocular disease is a dry eye disorder that affects the surface of the eye. The disease includes also known as keratoconjunctivitis sicca, keratitis sicca 'sicca syndrome' xerophthalmia, tear film disorder, tear fluid Decreased tear production, aqueous tear deficiency, and the condition of the Meibomian gland dysfunction. Dry eye syndrome may include types associated with SjGgren's syndrome (SS), such as Sjogren's syndrome associated with dry keratoconjunctivitis (SjSgreiVs syndrome 111 201217528 associated keratoconjunctivitis sicca), but may also include Non-Sjdgren's syndrome associated keratoconjunctivitis sicca ° The disease may or may not Other manifestations of autoimmune diseases. IL-1 is involved in the pathogenesis of dry eye syndrome. See, for example, Enriquez de Salamanca et al. 2010, Mol. Vis., 16: 862-873. Individuals with a dry eye condition may present with an eye dry inflammation and may experience scratchy 'stingy', 'itchy', burning or pressured sensations, Irritation 'pain', and redness. Dry eye can be associated with both excessive eye watering and, in contrast, tearing production. A receptor binding agent can be administered to these individuals to improve or avoid the onset or worsting of one or more of these symptoms. A receptor binding agent can also be used to relieve pain in individuals experiencing pain, such as, for example, eye pain due to neuroinflammation. In some embodiments, the ocular disease is an ocular disease associated with a systemic autoimmune disease (such as SjSgren's syndrome and wind diffuse arthritis), or with IL-1 or Other IL-1 interleukin family members are associated with diseases. The patient may or may not have _ systemic autoimmune disease or other manifestations of systemic autoimmune disease. A receptor binding agent can also be used to treat other conditions that affect the surface of the eye (such as 112 201217528 is the cornea). These diseases include corneal ocular surface inflammatory conditions 5 corneal neovascularization, keratitis, including peripheral ulcerative keratitis and bacterial keratitis (microbial) Keratitis). The receptor binding agent can be used to treat an individual who has undergone corneal wound healing (e.g., an individual with a corneal wound). A receptor binding agent can be administered to a dose that is being accepted, is undergoing, or is being involved in the eye, for example: corneal transplantation / keratoplasty, keratoprosthesis surgery , lamellar transplantation, individuals recovering from the process of selective seiective endothelial transplantation. See, for example, Dana, 2007, Trans Am Ophthalmol Soc, 105: 330-43; Dekaris et al., 1999, Current Curr Eye Res, 19(5): 456- 9; and Dana et al., 1997, Transplantation, 63: 1501-7. A receptor binding agent can be used to treat diseases affecting conjunctiva, including conjunctival scarring disorders and conjunctivitis. The receptor binding agent can be used to treat other diseases such as pemphigoid syndrome and Stevens-Johnson syndrome. One of the receptor binding agents described herein can be administered to a body to modulate neovascularization in or around the eye. See, for example, Dana, 2007, Trans Am Ophthalmol Soc, 105: 113 201217528 330-43. A receptor binding agent can be administered to an individual having an allergic reaction that affects the eye, for example, undergoing a severe allergy (at 〇 pic) like allergic conjunctivitis. Individuals with eye diseases. For example, the receptor binding agent can be administered topically. See also, for example, 'Kane-MyersAM et al., 1999, Invest Ophthalmol Vis Sci, 40(12): 3041-6. A receptor binding agent can be administered to an individual having an autoimmune disease affecting the eye. Exemplary autoimmune ocular diseases include sympathetic ophthalmia, Vogt-Koyanagi Harada (VKH) syndrome, scattered catenary choroidal lesion (birdshot retinochoriodopathy), eye scar Ocular cicatricial pemphigoid, Fuchs' heterochronic iridocyclitis, and different types of uveitis. A receptor binding agent can be administered to a subject to treat any of the above diseases. A receptor binding agent can be administered to an individual who is at risk of or at risk of diabetic retinopathy. See, for example,

Demircan et al., 2006, Eye Journal (Eye), 20: 1366-1369 and Doganay et al., 2006, Eye Journal (Eye), 16: 163-170. Uveitis includes both acute and chronic types and includes inflammation of the iris, the ciHary body, and one of the choroids. Chronic types may be associated with systemic autoimmune diseases such as 'Behcet's syndrome', ankylosing spondylitis

114 201217528 (ankylosing spondylitis), juvenile rheumatoid arthritis, Reiteris syndr〇me, and inflammatory bowel disease. In anterior uveitis, the inflammatory response occurs mainly in the iris (also known as iritis). Anterior uveitis can affect individuals with systemic autoimmune diseases, but can also affect individuals who do not have systemic autoimmune diseases. Intermediate uveitis involves the anterior vitreous, peripheral retina, and inflammation of the ciliary body, usually accompanied by a few anterior or chorioretinal Inflammation. Pan planitis is caused by inflammation of the pars plana between the iris and the choroid (ch〇r〇ids). Posterior uveitis is involved in the uveai tract and mainly choroids, also known as choroiditis (ch〇r〇iditis). Posterior uveitis can be associated with a systemic infection (systemic infecti〇n) or an autoimmune disease. It can last for months or even years. A receptor binding agent can be administered to a body to treat any of the aforementioned types of uveitis. See also 'For example, Tsai et al., 2009, Mol Vis' 15 : 1542-1552 and Trittibach P et al., 2008, Gene Ther. 15(22) : 1478-88 . In some embodiments, a receptor binding agent is used to treat an individual at risk of or suffering from age-related macular degeneration (AMD). The receptor binding agent can be administered topically to the eye's injection (e. g., 'intravitreally) or systemic 115201217528 (provided systemically). See, for example, 〇丨s〇n et al., 2009, 〇cui Immun〇1 Inflamm, 17(3): 195-200. One of the receptor binding agents described in the present invention can be administered by any mode for treating ocular diseases. The agent can be delivered by a parenteral mode. Alternatively or additionally, the agent can be delivered directly to the eye or to the vicinity of the eye. For example, the protein can be administered topically or intraocularly, for example, as described below. Ocular Delivery Formulations and Methods Ophthalmic formulations containing a styling binder can be delivered as a topical administration, for example, as a droplet (liquid drop) or ointment administration, or implantation, for example, into the anterior chamber or conjunctival sac. The droplets can be delivered using an eye dropper. When formulated as an ocular delivery, the receptor binding agent can be 0.0001 to 0.1%, 0.001-5%, for example, 〇.〇〇5~〇.5%, 0.05~0.5%, 0.01~5°/〇' A concentration of 0_1~2% or 1%~5°/〇 exists. Typically the ophthalmic formulation is applied directly to the eye 'including topical application to eyelids or instillation into the void between the eyeball and the eyelids (cul-de) -sac)). The ophthalmic formulation can be designed to rapidly mix with lacrimal fluids and spread over the surface of the cornea and conjunctiva. Using the usual technique of administration, the majority portion of the drug will be deposited underneath.

S 116 201217528 low ( (lower fornix). Capillarity, diffusional forces, and blinking reflex drive the drug into the precorneal film, through which the drug penetrates and passes through the cornea. The ophthalmic formulation may also include one or more other agents, such as an anti-inflammatory steroid, such as rimexolone, loteprednol, and meridsone. ) with hydrocortisone, or a non-steroidal anti-inflammatory. For example, the steroid may be present at a concentration of from 0.001 to 1%. In some embodiments, no steroids are present. For example, the receptor binding agent is the only active agent in the formulation. The formulation may also comprise one or more of the following ingredients: surfactants, tonicity agents, buffers, preservatives, co-solvents and adhesives. Viscosity building agents. Tonicity agents can be used to adjust the tonicity of the composition, such as the tension of natural tears. For example, potassium chloride, sodium chloride, magnesium chloride, calcium chloride, dextrose and/or mannitol It can be added to achieve a suitable tonicity, for example, physiological tonicity. Tonicity agents can be added in an amount sufficient to provide an osmolality of about 150 to 450 milliosmoles (mOsm) or 250 to 350 milliosmoles (mOsm). 117 201217528 This formulation may also include a buffer suitable for ophthalmic denvery. The buffer may include - a buffer component (for example: sodium phosphate, sodium acetate (four) out (four) coffee (four) 'sodium citrate' sodium borate (s〇dium b〇rate) or boric acid (b 〇ric acid)) 'Specially, under storage conditions, it has a buffering effect on pH changes. For example, the buffer may be selected such that the target pH falls within the range of pH 6.0 to 7.5, such as 6.5 to 7.5. The 6 Hai formula may include an aqueous (aque〇us) or phospholipid cairiers. Particularly for the treatment of dry eye, the formulation may include agents that provide short-term relief, such as compounds that lubricate the eye and aid in tear formation. For example, a phospholipid carrier (which includes one or more phospholipids) can be used to provide short-term relief. Can be used as an example of artificial tears carriers or artificial tears compositions including teardrop eye drops (Tears)

NaturaleTM) (Aercom, Inc., Texas, USA (Alc〇n Labs, Inc., TX USA)) commercial product. For example, in milliliters, the formulation may include: 1 milligram (mg) of dextran, 70 and 3 milligrams (mg) of hydroxypropyl methylcellulose, and optionally (optionally) ) — Preservative, such as POLYQUAD® (polyquaternium-1) 0.001% (m/v). Examples of lipid carrier formulations include those disclosed in U.S. Patent No. 4,804,539 (U.S. Patent No. 4,804,539), U.S. Patent No. 4,883,658 (1, 4, 883, 658), U.S. Patent No. 5,075,104 (U.S. Patent No. 5,075 , 104), U.S. Patent No. 5,278,151 (US 5,278,151), and U.S. Patent No.

S 118 201217528 5,578,586 (US 5,578,586). The formulation may also include other compounds which act as a lubricant or wetting agent. These include viscosity agents such as monomeric polyols such as glycerol, propylene glycol, ethylene glycol, and polymeric polyols. Polyols, such as polyethylene glycol, a variety of polymers in the cellulose family: hydroxypropylmethyl cellulose ("HPMC") 曱 曱 曱 cellulose sodium (carboxy Methylcellulose sodium), hydroxy propylcellulose ("HPC"), dextrans, such as dextran 70; water soluble proteins, such as gelatin Gelatin); with vinyl polymers, such as polyvinyl alcohol, polyethylene, B, _ (polyvinylpyrrolidone), povidone and carbomer 'Like carbomer 934P (carbomer 93 servant), carbomer 941 (carbomer 941); carbomer 940 (carbomer 940), carbomer 974P (carbomer 974P). Other examples include polysaccharides such as hyaluronic acid and salts thereof and chondroitin sulfate and salts thereof, and acrylic acid polymers. In certain embodiments, the formulation has a viscosity value between 1 and 400 centipoise (cP). The formulation can be packaged as a single or multi-dose use 'eg, packaged in a bottle with an associated dropper or packaged in a group ( a set of) Single 119 201217528 Use (single-use) droppers. The formulation may include one or more preservatives, for example, to avoid microbial and fungal contamination during use. Exemplary preservatives include: benzalkonium chloride, chlorobutanol, benzododecinium bromide, methyl paraben, Propyl paraben, phenylethyl alcohol, edetate disodium, sorbic acid, and polyquaternium-1 And the content thereof may be a concentration of 0.001 to 1.0% w/v. It is also possible to provide a formulation comprising a receptor binding agent which is sterile and free of preservatives. This formulation can be prepared as a single use application. Ophthalmic packs can be used to extend the contact time between the eye and the ophthalmic formulation. A cotton pledget is filled with the formulation and then inserted into the superior or inferior fornix. A receptor binding agent can also be administered by means of iontophoresis. This procedure keeps the solution in contact with the cornea on an eyecup with an electrode. The diffusion of the drug is affected by a difference in electrical potential. A receptor binding agent can also be delivered by injection, for example, subconjunctival injection. The formulation can be injected under the conjunctiva to facilitate passage through the sclera and into the eye by simple diffusion. The formula can also be injected under the conjunctiva and underneath the eye.

120 201217528 The capsule (Tenon's capsule) is delivered to the ciliary body 'choroid', and the retina. The formulation can also be administered by retrobulbar injection. For dry eye and other surface disorders, individuals can use one or more of the following methods to evaluate: Ocular Surface Disease Index (OSDI), corneal and conjunctival staining, and Schirmer test. The Ocular Surface Disease Index (OSDI) is a 12-item questionnaire that provides a rapid assessment of ocular irritation symptoms consistent with ocular surface inflammatory diseases, including DES, and its equivalent to vision. The effect on the related function (visi〇n_related functioning). See, for example, the Journal of Ophthalmology and Inflammology (〇cul Immunol Inflamm)' 2007, Sep-Oct; 15(5): 389-93. The 12 projects of the shai OSDI questionnaire were graded according to the scale of 4 (graded). Scores are generated based on responses to provide a 0 to 100 graded OSDI score, with higher scores representing greater disability. From the baseline, the negative change represents a visually relevant function (visjon_reiated functi〇n) and an improvement in ocular inflammatory disease. Corneal and Conjunctival Staining: Corneal staining is a kind of epithelial disease (epitheliaidisease) or epitheiial barrier rupture (can be seen on the surface of the eye like dry eye) Measuring method of inflammatory disease). Even if there is no obvious dry eye on &a bed (clinjcally eviden.t dry 121 201217528 eye) 'If there is significant lid disease (such as posterior blepharitis), corneal staining can be presence. Corneal staining is highly associated with ocular discomfort in many patients, although not all patients; as noted above, corneal staining is generally associated with 〇SDi. For corneal fluorescein staining, a tear film can be stained with a saline-moistened fluorescein strip or a 1% sodium fluorescein solution. The entire cornea is then subjected to slit-lamp evaluation with a yellow barrier filter (#12 Bratten) and cobalt blue illumination. To test (examined). The staining was scored according to the Oxford Schema. Conjunctival staining (c〇njunctivai staining) is similarly a method for measuring epithelial disease or break in the epithelial barrier of the ocular surface. Conjunctival staining was performed under lissamine using lissamine green. Use a Saline-moistened strip or a 1% Hssamine green solution to stain the tear film, and then at a time greater than 3 secs but less than 2 Between the flat and the inter-conjunctival staining (interpaipebrai conjunctival staining). White light with moderate intensity was used, and the 基xford Schema was used to perform only the interpa丨pebral region of the nasal and temporal conjunctiva staining. score. Schirmer Test: The Simer test is based on the presence of a negative filter-paper strip (Whatman #41 (filter paper), 5 x 35 mm (strip) strips were taken at the inferior cul-de-sac. The test was conducted in a dimly lit room. The patient gently closed his/her eyes for five minutes and removed the strips. Since the tear strip is removed from the eye, the tear front will continue to advance millimeters, so the tip of the tear is marked with a ball-point pen at the exact fifth minute. Aqueous tear production was measured by the length of the millimeter of the strip soaked during a five minute period. In the absence of anesthesia, the result of the Himer test is 10 mm (mm) or less and in the case of anesthesia, the result of the Himer test is 5 mm (mm) or less is considered abnormal. . One of the positive changes from baseline indicates an improvement in one or more of the ocular inflammatory conditions described herein. Formulations and Methods of Pulmonary Delivery A receptor binding agent can be formulated for inhalatory or other modes of pulmonary delivery, for example, administration of the agent. Tissue to the respiratory tract, for example, upper and lower respiratory tract. Three common systems that can be used to locally deliver agents to the pulmonary air passages include dry powder inhalers (DPIs), metered dose inhalers (MDIs). With nebulizers. Dosing inhalers (MDIs) can be used to deliver receptor junctions in a soluble form or as a dispersion 123 201217528 Mixture. Typically, metered dose inhalers (MDIs) include a Freon or other propellant relative to a vapor pressure that pushes the spray drug at the start of the 3 liter device (aer 〇s〇iize(j medication) enters the respiratory tract. Conversely, dry powder inhalers (Dpis) usually rely on the patient's inspiratory efforts to introduce a dry powder form of medicine into the lungs. Nebu丨izers form a medical aerosol to be inhaled by transferring energy to a liquid bath. The ahai agent can be one; the east dry form (iy0phiiized f〇rm) (for example) Stored at room temperature and before inhalation (inhaiatio...reconstituted into solution. Use of a fluoride medium during liquid ventilation or pulmonary lavage ( Feuor〇chemicai medium) Direct pulmonary delivery of the drug is also a possible delivery mode. These and other methods can be used to deliver a receptor binding agent. The dosage form is in the form of at least about 〇.〇2, (U, 0.5, 1, 1.5, 2, 5, 10, 20, 40 or 50 mg / squirt once (mg/puff) or more ( The dosage unit is delivered. The sigma body binder can conveniently be in the form of aerosin spray presentation from pressurized packs or nebulizers. The use of a suitable propellant, such as 'dichlorodifluoromethane, trichlorofluoromethane' dielilorotetrafluoroctliane, dioxide or other suitable gas, is delivered. In the case of a pressurized aerosol, the dosage unit can be delivered by providing a valve (vaWe)

124 201217528 Send a certain amount to determine. Capsules and cartridges for an inhaler or insufflator can be formulated to contain the receptor binding agent with a suitable powder base (eg, lactose). Or a powder mix of starch, if the granule is a formulated granule). In addition to the formulated or unformulated receptor binding agent, other materials (such as 100% DPPC) or other surfactants can be mixed together to facilitate the formulation or unformulated Delivery and dispersion of body binding agents. The particle size can also be varied to control whether this should be delivered to the lower or upper respiratory tract. For example, those having a particle size ranging from 1 to 5 microns or from 1 to 50 microns may be used for the lower or upper respiratory tract, respectively. Delivery enhancers (e.g., surfactants) can be used to further enhance pulmonary delivery. A surfactant is generally a compound having a hydrophilic and lipophilic moiety that interacts with one of the two immiscible phases to promote absorption of a drug. . There are several reasons why surfactants contribute to dry particles, for example, reducing particle agglomeration and reducing phagocytosis of macrophage cells. Surfactants are well known in the art and include phosphoglyceres, for example: phosphatidylcholines 'L-alpha-phosphatidylcholine dipalmitoyl (DPPC) with diphosphatidyl glycerol (DPPG); hexadecanol; fatty acids; polyethylene glycol (polyethylene 125 201217528 glycol) (PEG); polyoxyethylene-9 -(polyoxyethylene-9-) ; aurylil (auryl ether); palmitic acid; oleic acid; sorbitan trioleate (Span 85); glycocholate (glycocholate); surfactin; poloxomer; sorbitan fatty acid ester; sorbitol gf glycerol sorbitan trioleate; tyloxapol ; with lipid lipids (phospholipids). The present invention is also characterized by specifically identifying antibodies to one of the chimeric intercellular regions described herein. For example, these antibodies prefer to bind to a chimeric region relative to any parental interferon region. For example, a specific antibody can bind to an epitope comprising a junction between a segment of a first parent cell and a second parent cell. Exemplary Second Agents One of the receptor binding agents described herein can be administered with a second dose. These two doses can be co-administered or administered separately, for example, using different regimens. The second agent exemplified includes an anti-inflammatory agent. In one embodiment, the second agent is an IL-17 antagonist (including all antagonists of IL-17 family members, for example: IL-17A 'IL-17F, IL-17B, IL-17C 'IL-17D, an antagonist of IL-17E). Exemplary IL-17 antagonists include: binding to IL-17 (including IL-17A, IL-17F 'IL-17B, IL-17C, IL-17D, and IL-17E) and antagonizing IL-17 Signal-transmitting agents (eg, antibodies and other binding proteins); binding to one or more IL-17 receptors that are mediated by δ 126 201217528, such as IL-17RA and IL-17RC 'and antagonizing IL-17 Agent (eg, an antibody to other binding proteins); binds to one comprising IL_丨7 and at least one acceptor subunit, eg, IL-17 and I1-17RA, or IL-17, IL-17RA, and IL -17RC, a complex that antagonizes the signaling of il-17 (eg, antibodies and other binding proteins); and agents such as soluble receptors, which include one or more of IL-17RA and IL- The soluble extracellular region of 17RC antagonizes the signalling mediation mediated by IL-17. In another embodiment, the second agent is an IL-12 antagonist. Exemplary IL-12 antagonists include: agents that bind to IL-12 (including p35 and p40) and antagonize signaling by IL-12 (eg, antibodies and other binding proteins); bind to one of IL-12 Or a plurality of receptors, such as iL-12Rpl or IL-12RP2, and antagonizing the signaling of IL-12 (eg, antibodies and other binding proteins); binding to a unit comprising P35 'p40 and at least one receptor (subunit), for example, a complex of il-12RP1 or IL-12R02, and an agent that antagonizes signaling by IL-12 (eg, antibodies and other binding proteins); and agents such as soluble receptors, including One or more of the soluble extracellular regions of IL-12ΙΙβ2 and antagonize the signal transduction mediated by IL-12. In another embodiment, the second agent is an IL-23 antagonist. Exemplary IL-23 antagonists include: an agent that binds to IL-23 (including pl9 and p40) and antagonizes signaling by IL-23 (eg, antibodies and other binding proteins); binds to one of IL-23 Or a plurality of receptors, such as IL-12RP1 or IL-23R, and antagonizing the signalling agent (eg, antibodies and other binding proteins) mediated by il-23; binding to a pl9, p40 and at least one receptor subunit 127 201217528 (subunit), for example, a complex of IL-12R01 or IL-23R, and an agent that antagonizes signaling by IL-23 (eg, antibodies and other binding proteins); and agents such as soluble receptors, It includes one or more soluble extracellular regions and antagonizes the signal transmission mediated by -23. Examples of IL-23 are not described. See, for example, 'Beyer et al., J. Mol. Biol., 2008, doi: 10.1016/j.jmb_2008.08.001. Animal Models A receptor binding agent can be evaluated in an animal model for human disease (e.g., a human autoimmune and/or human inflammatory disease). This agent is specific to the corresponding target protein in animals. Rheumatoid Arthritis Models. A receptor binding agent can be evaluated in an animal model of rheumatoid arthritis, for example, a collagen-induced arthritis (CIA) model. See, for example, Mclndoe et al., 1999, Proc. Natl. Acad. Sci. USA, 96: 2210-2214; Issekutz, A. C. et al., Immunology , 1996, 88: 569; and "Current Protocols in Immunology", Unit 15.5 (Unit 15.5), Coligan et al. (eds.), John Wiley & Sons, Inc. This model was established by immunization of rat/mice sensitive strains with native type II collagen. Collagen in Freund's complete adjuvant (Complete Freund's s 128 201217528

Adjuvant) (CFA) emulsified and intradermally (100 micrograms (μβ) collagen: ι00 μg (side) CFA/mouse) at the base of the tail (at the base of the tail) ). Control group mice were injected intradermally with 0.05 ml (ml) of steamed water/CFA emulsion. A booster injection of collagen in incomplete adjuvant was administered 21 days after the primary immunization. The disease is caused by an autoimmune reaction induced by the immunological action of collagen. Using a defined scale, joints can be scored for arthritis, inflammation, pannus, cartilage damage, and bone resorption. For example, the severity of arthritis can be scored according to the following criteria: 0 = no visible effects of arthritis; 1 = - refers to (〇ne digit) or joint edema ( Edema and erythema; 2 = edema and erythema of two joints; 3 = edema and erythema of more than two joints; 4 = full palm Severe arthritis with the entire paw and digits, along with the ankle of the ankle and the deformity of the limb. The scores for each limb can be added up and recorded as the arthritic index (AI) of each individual animal. Other scoring schemes can also be used for these or other standard criteria. Multiple Sclerosis. Experimental allergic encephalomyelitis (EAE) is a useful murine model of multiple sclerosis (murine 129 201217528 model). A receptor binding agent can be evaluated in this EAE model. eaE is an autoimmune disease intervened by a T cell characterized by inflammation of tau cells and mononuclear cells followed by demyelination of axons in the central nervous system. (See, for example, Bolton, C., 1995, Multiple Sclerosis (Muitiple)

Sclerosis), 143 (Bolton, C., 1995, Multiple Sclerosis, 143). The illustrated operating manual can be found in the Immunology Experimental Operation Manual (Current)

Protocols in Immunology)" Units 15.1 and 15.2 (Unit 15.1 and 15.2); Coligan et al. (ed.), John Wiley & Sons, Inc. Models for myelin disease have also been available. In this model, oligodendrocytes or Schwann cells are grafted into the central nervous system, for example , as described in Duncan et al., 1997, Molec. Med. Today, 554-561. Allograft. A receptor binding agent can be evaluated in an animal model of skin allograft rejection, e.g., using murine tail-skin grafts. Skin allograft rejection is mediated by T cells, helper T cells and killer-effector cells. See, for example, "Current Protocols in Immunology", Unit 4·4 (Unit 4.4); Coligan et al. (eds.), 1995, published by John Wiley & Sons, Inc. Other transplant rejection models can also be used. See, for example, Tinubu et al., 1994, Journal of Immunology (J.

S 130 201217528

Immunol.), 4330-4338 ° Inflammatory bowel disease (IBD) and colitis (Colitis) model. An exemplary model of inflammatory bowel disease is the use of a model that transfers CD4+CD45Rb-high (CD4+CD45Rb-high) cells to severely combined immunodeficiency mice (SCID mice). See, for example, Hirano et al., Journal of Pharmaceutical Sciences (Pharmacol Sci), 2009, June (Jun); 110(2): 169-81, and IL-10 deficient mice genetically transgenic (transgenic IL) -10 deficient mice) use. See, for example, 'Inaba et al., Inflamm Bowel Dis., DOI: 10.1002/ibd. 21253, 2010. Another exemplary colitis model uses dextran sulfate sodium (DSS) to induce acute colitis. For example, by 5% (weight/volume) (wt/vol) DSS (molecular mass 30-40 kDa; ICN Biomedicals, Aurora, Ohio) in free (ad libitum) drinking water State (OH)) is administered to induce colitis in mice. Symptoms caused by such treatment are bloody diarrhea, weight loss, colon shortening and mucosal ulceration accompanied by neutrophil infiltration. The histological feature of DSS-induced colitis is that inflammatory cells invade the lamina propria, accompanied by lymphoid hyperplasia, local duct damage (focal crypt) Damage), and epithelial ulceration. These changes are thought to expand because of the toxic effects of DSS on epithelial tissues and the phagocytosis and production of TNF-α and IFN-γ in the intrinsic 131 201217528 丨amina propria cells (production of TNF) -alpha and IFN-gamma). See, for example, Hassan et al., Public Science Library · PloS One ’ 2010, January (Jan) 25; 5(1): e8868. Dry Eye Disease Modes. A receptor binding agent can be evaluated in a mouse model of dry eye disease. Dry eye syndrome in mice can be induced by subcutaneous injection scopolamine, and the mice are then placed in a controlled-environment chamber. As a specific example, a normal healthy 6 to 10 week old mother C57BL/6 mouse can be induced to have dry eye syndrome by continuous exposure to a dry environment in an environmentally controlled chamber. The chamber has a low relative humidity of less than 30% (generally about 19%), a high airflow (15 liters/minute) and a strange temperature (constant temperature) (about 22 ° C). The mice placed in the chamber can also be treated with scopolamine to inhibit tear secretion. Sustained-release transdermal scopolamine patches are available from Novartis (Summit, N.J.). A quarter of a patch can be applied to the depilated mid-tail portion of the mouse every 48 hours. The combination of a contr〇iied environmental chamber and a scopolamine produces severe dry eye in a relatively short period of time (about 2 to 4 days). The controlled environmental chamber can be used according to 132 201217528

Barbino et al., Ophthalmology Research and Visual Mechanics Journal (11^631; .0卩1^1^1.'\/^· Sci.), 46: 2766-2711, prepared in the 2005 paper, And can control air flow, humidity, and temperature. The symptoms of dry eye syndrome in mice can be monitored, for example, by performing: a) cotton thread test to measure aqueous tear production 'in patients with dry eye syndrome Usually reduced; b) corneal fluorescein staining, which is a marker of corneal surface damage; and general ophthalmic examination ° cotton test (Cotton) Thread Test): Cotton thread test can be used to measure tears by phenol red (Zone-Quick, Lacrimedics, Eastsound, Wash.) (Zone-Quick, Lacrimedics, Eastsound, Wash.) produce. Under a magnifying fluorescent lamp, the thread is held by a jade robot (jeweler f〇rceP) and placed outside the conjunctival fornix. The lateral cantus is 30 or 60 seconds. The tear distance (in millimeters (mm)) was read under a microscope using a hemacytometer scale. Corneal Fluorescein Staining: Corneal fluorescence staining can be performed by applying 1.0 microliter (ul) of 5% fluorescein to the conjunctival sac under the eye using a micropipette (micropipette) Inferi〇r conjunctival sac) was evaluated by s. After the fluorescein was instilled for 3 minutes, the cornea was examined using a slit light biomicroscope (Slit lamp 133 201217528 biomicroscope) using a cobalt blue. Utilizing a standardized National Eye Institute (NEI) grading system (with grades 0 to 3), for each of the corneal surfaces that are divided into five regions, in a blinded manner ( In a masked fashion) Punctuate staining. Diagnostic and Other Uses One of the receptor binding agents described in this case can be used to detect IL-1R1 in a sample, or cells expressing this receptor. For example, the agent may be labeled, directly or indirectly, with a label, or a signal, such as an enzyme, A radiolabel, an epitope, or a fluorescent protein (such as a green fluorescent protein). The agent can be contacted with a sample or cell to determine if the receptor is present in or on the cell, for example, by immunoblotting, immunofluorescence, and immunoassay. Enzyme immunoassay (ElA), radioimmunoassay (RIA) 'fluorescence energy transfer, Western protein, and other diagnostic and detection techniques. The receptor binding agent can also be labeled for in vivo tear detection and administration to a subject. The individual can be imaged, for example, by nuclear magnetic resonance (NMR) or other tomographic means. For example, the binder can be a radiolabel (eg, 1311, lllln, 1231, 99mTc, 32P, 1251, 3H, 14C, and 188Rh), 134 201217528 fluorescent labels (eg, fluorescent) Fluorescein and rhodamine, nuclear magnetic resonance active labels, can be scanned by a positive radiography (p0sitr〇n emission tomography) ("PET" scanner Detection of positron emitting isotopes, chemiluminescers (such as luciferin), and enzymatic markers (such as peroxidase or phosphatase) And the agent can be labeled as a c〇ntrast agent such as paramagnetic agents and ferromagnetic or superparamagnetic (which mainly changes the T2 reaction) Labeled. A receptor binding agent can also be used to purify cells that express the receptor to which they bind. For example, the receptor binding agent can be coupled. Up to an immobilized support (eg, magnetic beads or a column matrix) and contacting cells that can express the receptor. The slab can be rinsed (eg, using a Physiological buffers), and the cells can be recovered from the support. A receptor binding agent can also be used to purify the soluble form of the receptor to which it binds. For example, a sample containing the soluble receptor The immobilized receptor binding agent can be contacted and then, for example, after rinsing, can be recovered from the immobilized agent. The following non-limiting examples further illustrate embodiments of the invention described herein. 201217528 Examples 1 (Example 1) A nucleic acid system encoding a protein having the amino acid sequence listed in Table 3 (below) is constructed in a PET vector containing a tau7 promoter And ampicillin (pET31 series) or kanamycin resistance genes (pET28 series) (EMD Chemical Company, Gibbstown City, New Jersey, USA), and performance. Examples of coding sequences that can be used for performance are provided in Table 4. The three cases shown in Table enemy bonded proteins (Exemplary chimeric proteins) SEQ ID NO: (SEQ ID NO :) P01 APVRSLAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLE EKIDVSFVQGEESNDKIPVALGIHGGKMCLSCVKSGDET RLQLEAVDPKNYPKKKMDKRFAFIRSDSGPTTSFESAAC 17 P02 APVRSLAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLE EKIDVSFVQGEESNDKIPVALGIHGGKMCLSCVKSGDET RLQLEAVDPKNYPKKKMEKRFVFNKIEINNKLSFESAAC 18 P03 APVRSLAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLE EKFSMSFVQGEESNDKIPVALGLKEKNLYLSCVLKDDKP TL〇LESVDPKNYPKKKMEKRFVFIRSDSGPTTSFESAACP 19 P04 APVRSLAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLE EKFSMSFVQGEESNDKIPVALGLKEKNLYLSCVLKDDKP TLQLESVDPKNYPKKKMEKRFVFNKIEINNKLEFESAAC 20 P05 APVRSLNCRIWDVNQKTFYLRNNQLVAGYLQGPNVNLE EKFSMSFVQGEESNDKIPVALGLKEKNLYLSCVLKDDKP TLQLESVDPKNYPKKKMEKRFVFNKIEINNKLEFESAQFP 21 P06 APVRSLNCTLWDVNQKTFYLRNNQLVAGYLQGPNVEQQ VVFSMSFVQGEESNDKIPVALGLKEKNLYLSCVLKDDKP TLQLESVDPKNYPKKKMEKRFVFNKIEINNKLEFESAQFP 23 P07 APVRSLNCRIWDVNQKTFYLRNNQLVAGYLQGPNVNLE EKFSMSFVQGEESNDKIPVALGLKEKNLYLSCVLKDDKP TLQLESVDPKNYPKKKMEKRFVFNKIEINNKLEFESAQFP 24 136 201217528 Examples of the above-described code protein nucleic acid sequence shown in Table 4 series. In some embodiments, the nucleic acid sequence further comprises an ATG prior to the first nucleotide of the one listed below. In some embodiments, the nucleic acid sequence further comprises a stop codon (e. g., TAA, TAG, or TGA) following the last nucleotide of the underlying list. Table 4.1 illustrates the chimeric nucleic acid encoding the proteins (Nucleic acids encoding exemplary chimeric proteins) SEQ ID NO: (SEQ ID NO :) P01 GCACCTGTACGATCACTGGCCTTCAGAATCTGGGATGT TAACCAGAAGACCTTCTATCTGAGGAACAACCAACTA GTTGCTGGATACTTGCAAGGACCAAATGTCAATTTAGA AGAAAAGATAGATGTGTCCTTTGTACAAGGAGAAGAA AGTAATGACAAAATACCTGTGGCCTTGGGCATCCATGG AGGGAAGATGTGCCTGTCCTGTGTCAAGTCTGGTGATG AGACCAGACTCCAGCTGGAGGCAGTTGATCCCAAAAA TTACCCAAAGAAGAAGATGGACAAGCGCTTCGCCTTC ATCCGCTCAGACAGCGGCCCCACCACCAGTTTTGAGTC TGCCGCCTGCCCCGGTTGGTTCCTCTGCACAGCGATGG AAGCTGACCAGCCCGTCAGCCTCACCAATATGCCTGAC GAAGGCGTCATGGTCACCAAATTCTACATGCAATTTGT GTCTTCC 25 P02 GCACCTGTACGATCACTGGCCTTCAGAATCTGGGATGT TAACCAGAAGACCTTCTATCTGAGGAACAACCAACTA GTTGCTGGATACTTGCAAGGACCAAATGTCAATTTAGA AGAAAAGATAGATGTGTCCTTTGTACAAGGAGAAGAA AGTAATGACAAAATACCTGTGGCCTTGGGCATCCATGG AGGGAAGATGTGCCTGTCCTGTGTCAAGTCTGGTGATG AGACCAGACTCCAGCTGGAGGCAGTTGATCCCAAAAA TTACCCAAAGAAGAAGATGGAAAAGCGATTTGTCTTC AACAAGATAGAAATCAATAACAAGCTGAGTTTTGAGTC TGCCGCCTGCCCCGGTTGGTTCCTCTGCACAGCGATGG AAGCTGACCAGCCCGTCAGCCTCACCAATATGCCTGAC GAAGGCGTCATGGTCACCAAATTCTACATGCAATTTGT GTCTTCC 26 137 201217528 P03 GCACCTGTACGATCACTGGCCTTCAGAATCTGGGATGT TAACCAGAAGACCTTCTATCTGAGGAACAACCAACTA GTTGCTGGATACTTGCAAGGACCAAATGTCAATTTAGA AGAAAAGTTCTCCATGTCCTTTGTACAAGGAGAAGAA AGTAATGACAAAATACCTGTGGCCTTGGGCCTCAAGGA AAAGAATCTGTACCTGTCCTGCGTGTTGAAAGATGATA AGCCCACTCTACAGCTGGAGAGTGTAGATCCCAAAAAT TACCCAAAGAAGAAGATGGAAAAGCGATTTGTCTTCAT CCGCTCAGACAGCGGCCCCACCACCAGTTTTGAGTCT GCCGCCTGCCCCGGTTGGTTCCTCTGCACAGCGATGGA AGCTGACCAGCCCGTCAGCCTCACCAATATGCCTGACG AAGGCGTCATGGTCACCAAATTCACCATGCAATTTGTG TCTTCC 27 P04 GCACCTGTACGATCACTGGCCTTCAGAATCTGGGATGT TAACCAGAAGACCTTCTATCTGAGGAACAACCAACTA GTTGCTGGATACTTGCAAGGACCAAATGTCAATTTAGA AGAAAAGTTCTCCATGTCCTTTGTACAAGGAGAAGAA AGTAATGACAAAATACCTGTGGCCTTGGGCCTCAAGGA AAAGAATCTGTACCTGTCCTGCGTGTTGAAAGATGATA AGCCCACTCTACAGCTGGAGAGTGTAGATCCCAAAAAT TACCCAAAGAAGAAGATGGAAAAGCGATTTGTCTTCA ACAAGATAGAAATCAATAACAAGCTGGAATTTGAGTCT GCCGCCTGCCCCGGTTGGTTCCTCTGCACAGCGATGGA AGCTGACCAGCCCGTCAGCCTCACC AATATGCCTGACG AAGGCGTCATGGTCACCAAATTCACCATGCAATTTGTG TCTTCC 28 P05 GCACCTGTACGATCACTGAACTGCAGAATCTGGGATGT TAACCAGAAGACCTTCTATCTGAGGAACAACCAACTA GTTGCTGGATACTTGCAAGGACCAAATGTCAATTTAGA AGAAAAGTTCTCCATGTCCTTTGTACAAGGAGAAGAA AGTAATGACAAAATACCTGTGGCCTTGGGCCTCAAGGA AAAGAATCTGTACCTGTCCTGCGTGTTGAAAGATGATA AGCCCACTCTACAGCTGGAGAGTGTAGATCCCAAAAAT TACCCAAAGAAGAAGATGGAAAAGCGATTTGTCTTCA ACAAGATAGAAATCAATAACAAGCTGGAATTTGAGTCT GCCCAGTTCCCCAACTGGTTCCTCTGCACAGCGATGGA AGCTGACCAGCCCGTCAGCCTCACCAATATGCCTGACG AAGGCGTCATGGTCACCAAATTCTACATGCAATTTGTG TCTTCC 29 P06 GCACCTGTACGATCACTGAACTGCACGCTCTGGGATGT TAACCAGAAGACCTTCTATCTGAGGAACAACCAACTA GTTGCTGGATACTTGCAAGGACCAAATGTCGAGCAAC AAGTGGTGTTCTCCATGTCCTTTGTACAAGGAGAAGAA AGTAATGACAAAATACCTGTGGCCTTGGGCCTCAAGGA AAAGAATCTGTACCTGTCCTGCGTGTTGAAAGATGATA AGCCCACTCTACAGCTGGAGAGTGTAGATCCCAAAAAT TACCCAAAGAAGAAGATGGAAAAGCGATTTGTCTTCA ACAAGATAGAAATCAATAACAAGCTGGAATTTGAGTCT GCCCAGTTCCCCAACTGGTACATCAGCACCTCTATGGA AGCTGACCAGCCCGTCTTCCTGGGAGGGACCAAAGGC GGCCAGGATATAACTGACTTCACCATG CAATTTGTGTCT TCC 30 138 201217528 P07

GCACCTGTACGATCACTGAACTGCAGAATCTGGGATGT TAACCAGAAGACCTTCTATCTGAGGAACAACCAACTA GTTGCTGGATACTTGCAAGGACCAAATGTCAATTTAGA AGAAAAGTTCTCCATGTCCTTTGTACAAGGAGAAGAA AGTAATGACAAAATACCTGTGGCCTTGGGCCTCAAGGA AAAGAATCTGTACCTGTCCTGCGTGTTGAAAGATGATA AGCCCACTCTACAGCTGGAGAGTGTAGATCCCAAAAAT TACCCAAAGAAGAAGATGGAAAAGCGATTTGTCTTCA ACAAGATAGAAATCAATAACAAGCTGGAATTTGAGTCT GCCCAGTTCCCCAACTGGTTCCTCTGCACAGCGATGGA AGCTGACCAGCCCGTCAGCCTCACCAATATGCCTGACG AAGGCCAGGATATAACTGACTTCACCATGCAATTTGTG TCTTCC 31 may comprise such proteins from IL-Ιβ various IL-IRa of different residues, as described below. In the example Π (Π, P02, P03, P04, and P05, the interleukin region may have 48 to 70% of residues from IL-Ιβ and 55 to 78% of residues derived from IL-IRa. Since some amino acid residues are conserved between the two proteins, the sum of the degrees of identity for IL-Ιβ and IL-IRa can be greater than 100%.) Table 5 IL-Ιβ Residues (IL-Ιβ Residues) IL -IRA residues (Total residues) % IL-Ιβ % IL-IRA P06 130 62 152 85.5 40.8 P07 113 80 153 73.9 52.3 P05 108 85 153 70.6 55.6 P04 104 89 153 68.0 58.2 P03 94 99 153 61.4 64.7 P02 85 108 153 55.6 70.6 P01 74 119 153 48.4 77.8 Example 2 (Example 2) A protein containing a hexa-histidine tag, (hexa-histidinetag), at a concentration of 1 millimolar ( IPTG of mM) was induced in LB broth for 3 hours at 37 ° C, and was expressed in E. coli cell BL21 (DES) 139 201217528 strain. These cell lines were 20 to 50 mmol. Ear concentration (mM) tri-methylaminomethane (Tris) ' 0_5 molar concentration (M) sodium vaporized (NaCl), 2 · 5 millimolar concentration (mM) EDTA ' 0.1% Trit On Χ-100, lysed under conditions of pH 8.0. The lysate was pre-filled with a HiTrap® column (Singular Medical Equipment, Inc., Piscataway City, NJ, USA) (GE Healthcare,

Piscataway NJ, USA) performs IMAC chromatographic analysis (imaC chromatography). The protein was loaded at 20 millimolar (mM) sodium phosphate, 0.5 molar (Μ) sodium vaporized (NaCl) 10 millimolar (mM). Sit in imidazole, pH 7.4 buffer. It is at 200 millimolar (mM) concentration imidazole, 20 millimolar (mM) sodium phosphate, 0.5 molar concentration (M) sodium (NaCl), pH 7_4 The buffer is to be eluted. The proposed protein was dialyzed extensively in PBS, 0.1% Polysorbate 80, pH 7.4, concentrated using an Amicon Ultra® (l〇K) filter, and stored in 4. Or -80 ° C. Proteins lacking the hexa-histidine tag are purified by ion exchange chromatography. The P05 protein was purified by ion exchange chromatography. The lysate from the expressing cells is applied to a GigaCapSTM column at low pH (about pH 5.5) and without salts (conductivity of about 1 millisiemens per centimeter (mS/cm)). Tosoh Bioscience LLC, King of Prussia, PA, USA). The column is followed by a pH gradient (Buffer A = 10 millimolar (mM) acetic acid 140 201217528 (acetic acid) 'pH 5.5; buffer B (Buffer B) = 20 millimolar concentration (mM) Tris (tris) pH 8) is eluted. A 5 ml (ml) portion containing the priming protein (fracti〇n) followed by 5 ml (ml) of H20 and 5 ml (ml) of 20 mM (mM) trishydroxylaminomethane (Tris) pH 8) for dilution, and then applied to CaptoQTM resin (Singular Medical Equipment, Inc., Piscataway City, New Jersey, USA) (ge Healthcare, Piscataway NJ, USA) at a concentration of one millimolar (mM) A gasified sodium (NaCl) gradient to a concentration of 250 millimolar (mM) was eluted at 20 millimolar (mM) tris-hydroxyamino decane (Tris) pH 8.0. The protein was fully dialyzed and stored in 1.25 X PBS 0.1% TWEEN® 80 or 1.25 X PBS without TWEEN®. See the sixth picture. The P03 and P04 protein lines were purified by a similar method. Example 3 These proteins or supernatants containing such proteins can be used to evaluate IL-1 activity in a cell-based assay. HEK-BlueTM IL-Ιβ responsive cells are used to monitor IL-Ιβ activity (available from InvivoGen, Inc., San Diego CA, USA) . These cells include a SEAP reporter gene under the control of an IFN-β minimal promoter that binds to five NF-κΒ and five AP-1 binding sites. IL-Ιβ engagement to the IL-1 receptor on the cell surface leads to activation of NF-κΒ and production of SEAP. The SEAP report can be detected, for example, using QUANTI-BlueTM (InvivoGen, San Diego, California 141 201217528 Asia, USA (InvivoGen Inc., San Diego CA, USA)) and spectrophotometric analysis ). A HEK-Blue IL-Ιβ cell suspension can be prepared from cells cultured to 70-80% confluence. The resuspended cells were adjusted to have a concentration of ~330,000 cells/ml in fresh growth medium (culture medium: DMEM, 4.5 g/L (g/Ι) Glucose, 2 house molar concentration (mM) L-Glutamine, 10% (v/v) heat-inactivated fetal bovine serum (at 56 ° C) 30 minutes), 50 units/ml (u/ml) penicillin (penicillin), 50 μg/ml (pg/ml) streptomycin, 100 μg/ml (pg/ml) NormocinT). Reagents were added to a flat 96-well cell culture dish: 20 ng/ml IL-Ιβ 10 μl (ul) The agent was 10 microliters (ul), and the cell culture solution was 3 liters microliter (ui) to achieve a final volume of 50 microliters (ul). The positive and negative control (P〇sitive an (j negative controls) sample lines were prepared in parallel. Then the i5 〇 microliter (u丨) HEK-Blue IL-1 β cell suspension (~50,000 cells) was force Sigma was introduced into each well and the swarf plate was cultured overnight at 37 ° C in a 5% C02 tissue culture incubator (Tjssue cuiture incubator). In general, the final IL_丨β concentration (at 200 μl ( The final volume of ul) is 毫1 ng/ml (ng/ml). The activity of IL-Ιβ is evaluated every other day (after 12 to 15 hours). Before quantification 'according to the manufacturer's instructions (manufacturer's instruction) To prepare the UANTI-BlueTM s. Prepare a fiat bottomed 96-well assay plate and add 15 μl of liter to each well.

S 142 201217528 QUANTI-BlueTM solution. A conditioned medium from the well of the 96 well tissue culture plate, 50 microliters (ul), was added to each well of the assay disk. The tray was incubated at 37 ° C for about 15 to 20 minutes. The concentration of SEAP is then measured using a spectrophotometer at a wavelength of 620 to 655 nanometers (nm). result. As shown in Figure 7A, in this assay, the p〇6 protein acts as an IL-1RI agonist, the P07 protein acts as a partial agonist, and the P01 protein fails to act. In fact, the P01 protein acts as an antagonist when assayed in the presence of IL-Ιβ. Figure 7B shows the antagonism of IL-Ιβ activity by P01 at various IL-Ιβ protein concentrations using the HEKBlueTM cell assay described above. The antagonism increases as the amount of ροι increases (the X-axis reflects the microliters of the supernatant containing P01). Each of the proteins P01, P〇2, P〇3, P04 and P05 antagonizes the activity of IL-Ιβ. For example, see Figures 8A and 8B. The IC50 of p〇5 is less than about 5 ng/ml (ng/ml). P05 was tested for its ability to potentiate IL-1RI in this assay and was not observed to have any detectable agonistic activity, even at the highest concentration tested, 1 mg/ml (mg) /ml) ' Also. P01 ' P02 ' P03 ' P04 ' and P05 are also inhibited in MG-63 cells (a human osteosarcoma cell line that is reactive against jL-ΐβ) IL-6 beta-induced IL-6 expression . In the murine model of dry eye disease, the hexa-histidine-labeled Ρ05 line was observed to be biologically active. See also Example 8 below for the unmarked Ρ05. Example 4 (Example 4) A Reichert SR7000DC Dual Channel SPR (Dual Channel 143 201217528 SPR) system can be used to evaluate protein for soluble recombinant human IL-1RI (corresponding to IL-1RI) using surface plasmon resonance The binding properties of the extracellular region). This binding can be assessed in phosphate buffered saline with 0.005% Tween 20. The IL-Ιβ system was observed to have a KD value of one nanomolar concentration (nM) between 8 and 9 and a dissociation constant (Kd) between 2 and 3χ10·3 s·1. And in another experiment having a KD value of about 2 nanomolar (nM) and having an association constant of 1.3 to 1.5 χ 106 Μ···1, and about 2_9~3. 〇xl (a dissociation constant (Kd) of T3 s·1. See Figure 9. This Ρ01 protein binds to the binding kinetics similar to IL-Ιβ, but in this binding experiment The dissociation phase was not dissociated during the dissociation phase (about 180 seconds). Therefore, under similar conditions, the P〇1 protein binds to IL-1RI with an affinity higher than IL-1 β. IL-IRa binding It has been observed to have a value of about 0.33 nanomolar (nM), an association constant (Ka)' of about 1 χ, 1 and about 6. 6 χ 10-5 s·1 The dissociation constant (Kd). See Figure 9B. The chimeric interleukin region P〇1, P〇2, P03, P04, and P05 are A KD value having a range of about 12 to 1700 picomolar concentration (pM) was measured, and the range was about 3x104 m·1 S·1 to 3xl 〇6 M'V1, an association constant (Ka), and The range is about 2x10·5 to lxl〇-3 s-1, a dissociation constant (Kd). For example, see Figures 9C and 9D and Table 6 below.

S 144 201217528 Table 6 Protein ka (mV) Kd (s'1) Κ〇(ρΜ) IL-Ιβ 1.47 χ 106 M^s'1 2.95 χ 10_3 s·1 2010 IL-IRa 2_01 χ 105MV 6.58χ ΙΟ·5 S-1 326 P01 4.93 χ 104 2.32 χ 10_5 s·1 470 P02 3.39 χ 104 M^s-1 2.16 χ ΙΟ·5 s·1 636 P03 4.1 χ WM-V1 1.2 χ ΙΟ'3 s'1 290 P04 3.00 χ ΙΟ4 Μ·1 s·1 5.14 χ ΙΟ·4 s·1 1714 P05 3.47 χ 106Μ·ν 4.15 χ ΙΟ-5 s'1 12 P06 4.8 χ 106M_1s_1 1.7 χ ΙΟ'3 s'1 410 P07 1.58 χ ΙΟ4! ^·1 1.46 χ ΙΟ-3 s*1 92553 Example 5 (Example 5) Other exemplified chimeric IL-1 family proteins also include the following: P08 P09 P10 P11 P12 P13 P14

APVRSLAFRIWDVNQKTFYLKNNQLVAGYLQGPNVNLEEKFS MSFVQGEESNDKIPVALGLKEKNLYLSCVLKDDKPTLQLESV DPKNYPKXKMEKRFVFNKIEINNKLEFESAQFPNWFLCTAME ADQPVSLTNMPDEGVMVTKFYMQFVSS APVRSQAFR1WDVNQKTFYLRNNQLVAGYLQGPNVNLEEKFS MSFVQGEESNDKIPVALGLKEKNLYLSCVLKDDKPTLQLESV DPKNYPKKKMEKRFVFNK1EINNKLEFESAQFPNWFLCTAME ADQPVSLTNMPDEGVMVTKFYMQFVSS APVRSLAFRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKID VSFVQGEESNDKIPVALGLKEKNLYLSCVLKDDKPTLQLESVD PKNYPKKKMEKRFVFNKIE1NNKLEFESAQFPNWFLCTAMEA DQPVSLTNMPDEGVMVTKFYMQFVSS APVRSLNCRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKID VSFVQGEESNDK1PVALGLKEKNLYLSCVLKDDKPTLQLESVD PKNYPKKKMEKRFVFNKJEINNKLEFESAQFPNWFLCTAMEA DQPVSLTNMPDEGVMVTKFYMQFVSS APVRSLNCRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKFS MSFVQGEESNDKIPVALGLKEKNLYLSCVLKDDKPTLQLESV DPKNYPKKKMEKRFVFNKIEINNKLEFESAQFPNWFLCTAME ADQPVSLTNMPDEGVMVTKFTMQFVSS APVRSLAFR1WDVNQKTFYLRNNQLVAGYLQGPNVNLEEKFS MSFVQGEESNDKIPVALGLKEKNLYLSCVLKDDKPTLQLESV DPKNYPKKKMEKRFVFNKIEINNKLEFESAQFPNWFLCTAME ADQPVSLTNMPDEGVMVTKFYFQED APVRSLNCRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKFS MSFVQGEESNDKIPVAL GLKEKNLYLSCVLKDDKPTLQLESV DPKNYPKKKMEKRFVFNKIEINNKLEFESAQFPNWFLCTAME ADQPVSLTNMPDEGVMVTKFYFQED SEQ ID NO: 32 (SEQ ID NO: 32) SEQ ID NO: 33 (SEQ ID NO: 33) SEQ ID NO: 34 (SEQ ID NO: 34) SEQ ID NO: 35 (SEQ ID NO: 35) SEQ ID NO: 36 (SEQ [DNO:36) SEQ ID NO: 37 (SEQ ID NO: 37) SEQ ID NO: 38 (SEQ ID NO: 38) 145 201217528 The following polypeptide is an inlay region comprising at least two segments from IL-1 alpha and from At least two segments of IL-IRa.

SAPFSFLSNVKYNFMRIIKYEFRIWDVNQKTFYLRNNQLVAGY SEQ ID NO: 39 LQGPNVNLEEKFDMGAYKSSKDDAKITVILKISKTQLYVTAQD (SEQIDNO: 39) EDQPVLLKEMPEIPKTITGSETNLLFFWETHGTKNYFTSVAHP NLFLCTAMEADQPVSLTNMPDEGVMVTICFYLLENQA Example 6 (Example 6) can be connected by means of a peptide (Linker) sequence and select a new position for the end such that the N- terminus of the molecule Attached to the C-terminus to construct a circularly permuted IL-1 chimeric region. For proteins whose ends are derived from IL-Ιβ, the linker may be between 5 and 10 in length, for example, about 7 amino acids. The preferred position of the new terminus is located on the loop at the back of the receptor, for example, the β6-β7 loop (for example, corresponding to position 71 to 80 of SEQ ID No. 1 (SEQ ID ΝΟ: 1) The amino acid) or the β7-β8 loop (for example, an amino acid corresponding to positions 84 to 99 of SEQ ID No. 1 (SEQ ID ΝΟ: 1)). Examples of these IL-1 are arranged annular area of the fitting comprising: DKPTLQLESVDPKNYPKKKMEKRFVFNKIEINNKLEFESAQFPNWFL CTAMEADQPVSLTNMPDEGVMVTKFYMQFVSSGGSGGGSAPVRSL NCRIWDVNQKTFYLRNNQLVAGYLQGPNVNLEEKFSMSFVQGEESN DKIPVALGLKEKNLYLSCVLKD (SEQ ID NO: 40) (SEQ ID NO: 40) and NYPKKKMEKRFVFNKIEINNKLEFESAQFPNWFLCTAMEADQPVSLT NMPDEGVMVTKFYMQFVSSGGSGGGSAPVRSLNCRIWDVNQKTFY LRNNQLVAGYLQGPNVNLEEKFSMSFVQGEESNDKIPVALGLKEKNL YLSCVLKDDKPTLQLESVDPK (SEQ ID NO: 41) (SEQIDNO: 41) Example 7 (Example 7) Proteins P03, P04, P05, mIL-IRa (methionyl s IL-lRa), and IL-1β at pH 7.4 at 0.5 mg/ml (mg/ Ml) 146 201217528 Concentrate formulated in phosphate-buffered saline (PBS). The specific protein was combined with SYPRO orange dye (Invitrogen, Inc., California (invitr〇gen, CA)) at a stock concentration of 1:500 dilution and subjected to differential scanning fluorescence detection. Differential scanning fluorimetry. See, for example, 'He et al.', 2010, J. Pharm. Sciences, 99 1707-1720. Feuorescence measurements were monitored using an Agilent Agilent Mx3005 QPCR machine (Agilent Mx3005 QPCR machine) at temperatures ranging from 25 ° C to 95 ° C per minute rc. The Melting temperature (Tm) value is derived from the maximum value of the first derivative of the fluorescence transition. The protein P03 'P04, and P05 were observed to be unfolded at temperatures above 5 (TC and up to 59 °C, and have values above 59, 60, 62, and 64 °CiTm. Shown in Table 7 below and Figures 10A and 10B: Table 7 Protein (Protdn) TmCC) (0nset:fu^ng)Cc) mIL-IRa 56 48 IL-Ιβ 56 41 P03 65 59 P04 60 51 P05 65 59 P04 has a Tm value which is 4 ° C higher than the Tm values of mIL-IRa and IL-Ιβ, and exhibits a starting temperature of 3 ° C higher than ml L-1 Ra and about 10 ° C higher than IL-1 β. value. P〇3 and P〇5 have a T„^, which is more than mIL-IRa and IL-Ιβ 147 201217528

The Tm value is 9 ° C higher and exhibits a starting temperature value that is 11 ° C higher than mIL-IRa and about 18 ° C higher than IL-Ιβ. Example 8 (Example 8) Purified P05 (lacking the hexa-histamine marker) was formulated in 1.25 PBS and tested in a murine model of dry eye disease. In this model, female C57BL/6 mice from Jackson Laboratories, aged 6 to 10 weeks (with a 230% relative humidity, hydrogel food supplement) It was acclimated 1 to 2 weeks in an animal breeding room with envirodry enrichment enrichment before pre-screened on day 0 for fluorescent staining. For fluorescent staining, freshly prepared glomus was diluted in WFI H20 to 10 mg/ml (mg/mL), and 0.4 μl (μί) of each was administered in each eye. After 8 to 13 minutes of administration, the eye was scored using an Olympus fluorescent dissecting microscope. Utilizing a standardized National Eye Institute (NEI) grading system (with grades 0 to 3) for each of the corneal surfaces that are divided into five regions (fractions ranging from 0 to 15/eye) , record punctuate staining. Using a teaching bridge, two masked scorers flatten the mice at the same time s to give each eye a single collective score. Each eye score S7 ( The mouse with a maximum score of 15) was placed in a dry eye chamber on day 1 (20% ± 2% humidity and constant air flow was ~ 21 liters / minute / cage (L / min / cage)) 'And during the entire process of the experiment, it was held in the chamber by 148 201217528 (except when it was to be inspected). On day 3, the mice were again squashed and randomly assigned to a treatment group with 8 to 1 〇 mice (mice/gr〇Up). Mouse lines were randomly assigned such that each cage with 4 to 5 mice had approximately the same average disease score. Starting on the third day and after randomization treatment, the mouse line was locally administered with p〇5 or vehicle (i.25X PBS) in one eye drops to 3 μL/eye, twice daily (BID). The frequency. Mice were tested and subjected to the above-mentioned corneal fluorescence staining at 7, 9 and 11 days. The scorers were blinded to the treatment groups during the course of the experiment. Figure 11A is a bar graph of mean mean standard deviation (SEM) of mean corneal staining scores from two identical experiments performed on mice at 3, 7, 9, and 11 days. The following treatments were performed twice daily (bid): no treatment, vehicle (1.25X PBS) treatment, and treatment with 10 mg/ml (mg/ml) (1%) P05. P05 at 10 mg/ml (mg/ml) significantly reduced corneal staining at 7, 7, and 11 days of the experiment. Efficacy assessed by a reduction in corneal staining was also observed at concentrations as low as P05 (1 mg/ml). Recombinant IL-IRa produced in E. coli also moderately reduces corneal staining in this animal model. As shown in Fig. 11B, the effect of 10 mg/ml (mg/ml) of P05 was determined based on comparison with 10 mg/ml (mg/ml) of rat serum albumin in the same vehicle. 10 mg/ml (mg/ml) of rat serum albumin (MSA) had no effect compared to vehicle, and the effect of 10 mg/ml (mg/ml) P05 was compared to 1 mg/ml (mg/). The murine serum albumin system of ml) is statistically significant. As shown in Figure 11C, the 10 mg/ml (mg/ml) P05 line was also compared to 0.05% cyclosporine (Restasis®) in the 149 201217528 ophthalmic emulsion. P05 reduced corneal staining after one week of do dosing treatment, while no effect was observed with 0.05% cyclosporine ophthalmic emulsion. Example 9 (Example 9) P04 was purified and formed into a diffraction quality crystals at 20 ° C, 25% PEG 1500, 0.1 molar (M) PCB (pH 4.0). . The protein can be crystallized into a space group belonging to the 卩2, with a typical unit cell dimension of a = 44.5, b = 46.4, and c = 64.8. The crystals are diffracted to high resolution and extend to 1.47A. One of the datasets is attached to the Advanced Photon Source, beamline LS-CAT21ID-F (Chicago, Illinois, USA) ChicagoIL, USA)) was collected. The X-ray structure of P04 has been modeled by combining one of the relevant parts of the known IL-Ιβ and IL-IRa structures of the PDB structures 1ITB and 1IRA, and is solved by molecular replacement. Out (Vigers et al., 1997, Nature, 386: 190-194 and Schreuder et al., 1997, Nature '386: 194-200). The final model was refined to have a Rwcrk / Rfree value of 17.6% / 20.4% and contained a P04 molecule (140 residues) and 98 water molecules (Table 8). ). Residues 1 to 2, 48 to 49 and 85 to 93 of P04 are invisible in electron density and are missing in the final model.

S 150 201217528 Table 8 Crystallographic Data Collection and Refinement Statistics Data Collection Statistics Space group P212,2, Cell dimensions a,b ' c (A) 44.5 - 46.4 > 64.8 α'β, γ(.) 90 , 90 , 90 Wavelength (Wavelength) (A) 0.9787 Resolution (Resolution^ (A·) 50.0- 1.47(1.52- 1.47) •Emerge 0.039 (0.56) Ι/σΙ 37.7 (2.9) Completeness (%) 99.8 (100) Redundancy 6 (5.9) Refinement Statistics Resolution (A) 28.64-1.47 ( 1.53-1.47) No. of reflections 22926 R-work/Rfree 17.6/20.4 Average B value (A) B2.3 Rmsd bond lengths (A) 0.007 Mean square Rmsd bond angles'(°) 1.137 The number in parentheses corresponds to the highest resolution level and merge = [hid [Σί I li _ <1>I d] •^work = Zhk丨丨丨Fobs I _ I Fcalc 丨丨/Shki I ^"obs I where Fobs and Kale are observed and calculated Factorization (structure factors)

Rfree is calculated from a subset of such reflections (5%) that are not used for refinement. The P04 crystal structure has a fold similar to that predicted by simulation. A view of this structure is shown in Figure 1. The overlap of the two structures is shown in Figure 12A. The root mean square deviation (RMSD) of the IL-Ιβ and IL-IRa segments for the carbon chain backbone of the same residues on the respective parent molecules was 1.41A and 1.09A. P04 has at least one unique salt bridge and two unique hydrogen bonds, which are related to a residue derived from IL-1 β and one of the equivalent parts derived from IL-IRa Interactions: (1) Glu39-Lys64 (Glu39 from IL-IRA; Lys64 from IL-Ιβ), as shown in Figure 12B, (ii) Arg9-Glnl49 (Arg9 from IL-1RA; 151 201217528

Glnl49 is derived from IL-Ιβ) as shown in Figure 12C, and (iii) Serl52-Lys40 (Serl52 is derived from IL-1RA; Lys50 is derived from IL-Ιβ) as shown in Figure 12C. These unique interactions explain the increased thermal stability of P〇4 because these interactions do not exist in the IL-Ιβ and iL-IRa structures. These residues involved in these interactions are also present in p〇3 and p〇5, which also have increased thermal stability. Other embodiments are within the scope of the claims of the following claims. [Simple description of the drawing] Fig. 1 is a P04 structure diagram determined by X-ray crystal 丨〇graphic data. The backbone from the iL-lRa residue is shown in black, and the backbone from the IL-Ιβ residue is shown in gray. Figure 2 depicts three visual fields of a model in which the P〇5 protein binds to the extracellular region of human IL_1RI. Figure 3 depicts a model of a chimeric protein in which the IL_1Ra residue is depicted as black and the IL-1β residue is depicted as white. The protein depicted in the model: p〇1 (3ASI) 'Ρ03 (3rd map), Ρ〇4 (3C), Ρ〇5 (figure map), Ρ07 (3rd map), and Ρ06 ( Dijon map).

Figure 4 • Serving human IL-1 family interleukin-sequence alignment: Ιί-1β (SEQ ID NO: 1), IL_la (SEQ ID NO: 2 (SEQ ID N0: 2)) 'IL-IRa (SEQ ID NO: 3) ' IL-33 (SEQ ID NO: 4) (SEQ ID NO: 4), IL-36Ra (SEQ ID NO: No. 5 (SEQ ID NO: 5)), iL-36a (SEQ ID NO: 6), IL_36P (SEQ ID NO: 7 152 201217528 (SEQ ID NO: 7)), and IL-36y (SEQ ID NO: 8). The segments mentioned in the text of this case are identified in the sequence alignment. In addition, the β-folding and the loops between the flaps are also identified. The fifth panel is a list of one of the amino acid sequences of Ρ01 (SEQ ID NO: 17 (SEQ ID ΝΟ: 17)). Figure 5 is a list of one of the amino acid sequences of Ρ02 (SEQ ID NO: 18). Figure 5C is a list of the amino acid sequences of P03 (SEQ ID NO: 19). Figure 5D is a list of P04 (SEQ ID NO: 20) amino acid sequence. Figure 5E is P〇5 (SEQ ID NO: 21) List of one of the amino acid sequences of )). The segment from IL-Ιβ is shown in bold italics. See also Example 1 below. Figure 6 is an image of an SDS-PAGE colloid showing self-expression An exemplary sample of the protein purified from the E. coli cells of the receptor binding agent. The 15 and 20 kDa molecular weight marker lines are shown on the left side. The lanes are as follows: molecular weight marker (p.丨 and 6th lanes), extracts (2nd and 7th lanes), materials purified by cation exchange chromatography (3rd and 8th lanes), and anion exchange chromatography The material purified by the method (4th and 9th lanes), and the reduced sample of the material (4) uced Sample (5th and 1G lanes). The 2nd to 5th lanes are p()5 purified material' and the 6th to 10th lanes are the purified substances. See also example]. Figure 7A is a diagram showing the ability of the pm protein to be efficaciously transmitted by the pm protein relative to the IL-Ιβ and-negative control group (four) lysinase (gus) protein. Line 7B - depicts a graph of antagonism of IL-Ιβ caused by P01 at different concentrations of 153 201217528 IL-Ιβ. Figure 8A is a plot of IL-1 β (human) in the presence of 0 · 1 ng / ml (ng), from Ρ03 (hexa-histidine tagged), P04 (Ph6-histamine marker), P05 (via hexa-histidine marker), and IL-IRa antagonism of IL-Ιβ. Figure 8B is a plot of 0.1 ng/ml (ng/ml) of IL-Ιβ (human) in the presence of untagged forms of P01, P02, P03, P04, and P05 and IL. The antagonism of IL-Ιβ caused by the lysate of -IRa, and the estimated amount of protein used in the respective lysates. Figure 9 is a graph containing SPR data showing the binding kinetics of immobilized soluble IL-1RI with the following proteins (binding)

Kinetics): IL-Ιβ (Fig. 9A), IL-IRa (Fig. 9B), P〇4 (Fig. 9C), and P05 (Fig. 9D). Figure 10A is a thermal denaturation diagram of IL-1 Ra, il-1β, p〇3, Ρ04 and Ρ05 as described in Example 7. Figure 10 is a negative first derivative depiction of Figure 10A. Figure 11A is a bar graph showing the mean standard error (SEM) of the average score of the cornea, which is derived from the dry eye model of mice at 0, 3, 7, 9, and 11 days. In the two independent studies, the corneal fluorescence staining test results for each eye. The number of patients who did not receive any treatment in these mice was 18 (n - 18) '1 〇 mg/ml (mg/mi) p〇5 The number of treatments was 19 (n=19), or mxpBs, That is, the number of vehicle handlers is 20 (η = 20). The asterisk indicates the statistical significance of P05 relative to the vehicle, as shown below: * (Ρ < 0.05) and ** (Ρ < 〇 〇〇 5).

S 154 201217528 Figure 11B - a bar graph showing the data from the experiment of separation - showing the average angular 犋 staining score ± mean standard error (SEM) of the cornea of each eye - which is at 0th 3 ' 7 ' 9, mi days, a model of dry eye syndrome in mice. The number of those mice who did not receive any treatment was 80 = 8". The number of recipients receiving 1.25X PBS vehicle was 8 (n=8), and 1 mg/ml (mg/ml) of mouse serum albumin was accepted. The number of processors of murine senim albumin) (MSA) is 8 (η = 8), or the number of patients receiving ι〇mg/ml P05 is 9 (n=9). The asterisk indicates p〇5 relative to murine serum albumin. The statistical significance is as follows: * (P < 〇.〇5) and ** (p < 〇〇〇5). Figure 11C is a bar graph included in the UB map In the same experiment, the mice were treated with Restasis® (0.05% cyclosporine emulsion). The asterisk indicates the statistical significance of 1 >05 relative to Restasis®, as follows: * (p < 〇.05) and ** (p < 0.005). Fig. 12A depicts the structure of the X-ray crystal structure (black) of ρ 〇4 applied to a calculation model (gray) of its structure. The figure illustrates the interaction between K64 and E39 of P04; the 12th (: the interaction between K40 and R9 of p〇4 and the residues Q149 and S152 of C-terminal. [Key element symbol description] (none) 155

Claims (1)

201217528 VII. Patent Application Range: 1. An isolated protein comprising a chimeric interleukin-family cytokine region, wherein at least a first segment of the region is at least 2 amino acid lengths and is first One of the IL-1 family of interleukins has an amino acid identity of at least 80°/〇, and at least a second segment of the region is at least 2 amino acid lengths and is a first-IL-1 One of the family interleukins has at least 8% amino acid identity. 2. The protein of claim 1, wherein the first and second IL "family interleukins are selected from the group consisting of IL-Ιβ, IL-la, and IL_1Ra. 3 _如申The protein of the second aspect of the patent, wherein the first IL_丨 family interleukin is IL-Ιβ and the second IL-1 family interleukin is IL_1Ra. 4. According to claim 2 a protein, wherein the protein binds to il 1 receptor I and inhibits the activity of IL-1 receptor I. 5. The protein of claim 2, wherein the first segment of the interleukin region has The length of at least 20 or 30 amino acids is the same as that of one of IL-Ιβ. 6. The protein of claim 2, wherein the interleukin region comprises at least 80% identical to IL-1 Ra a residue of a portion corresponding to the residue and a residue of at least 80% of the residue corresponding to the corresponding residue on IL-Ιβ. 7. The protein of claim 3, wherein the interleukin region comprises (4) Amino acids identical to IL-IRa in the following positions: ARG11, SER13, GLN14, GLN15, GLU25, LYS27, LEU29, 156 201217528 HIS30 LEU31, GLN32, GLY33, GLN34, ASP35, MET36, GLN38, GLN39, ALA127, GLU128, ASN129, MET130, and GLN141 (according to IL-Ιβ numbering) and (b) amino acid equivalent to IL-Ιβ in the following positions: ALA1, PR02, VAL3, ARG4, LEU6, PHE46, GLN48, GLU5 SER52, ASN53, LYS55, ILE56, PR057, LYS92, LYS93, LYS94, LYS103, GLU105, ASN108, GLN149, PHE150, and SER152. The protein of the second aspect, wherein the interleukin region comprises the sequence of β-strand β2, β3, β10 and βΐΐ identical to IL-IRa and the sequence of β-strand β4′β6, β7, and β8 identical to IL-Ιβ. 9. The protein of claim 2, wherein the β1β2, β2β3, β8β9, βΙΟβΙΙ is at least 90% identical to one or more of the β11β12 loops from the corresponding loop from IL-IRa. The protein of claim 2, wherein the interleukin region comprises at least two discrete segments having a length of at least 20 amino acids, the segments being at least 90% identical to each other from IL-Ιβ. Section 11_If the patent application scope A protein of the first aspect, wherein the two discrete segments are identical to the respective corresponding segments from IL-Ιβ. 12. The protein of claim 1, wherein the two discrete segments are between 25 and 40 amino acid lengths. 13. A protein as claimed in claim 10, 1 or 12 wherein the amino acid which is not on the two discrete segments is at least 90% identical to Il-IRa. 14. The protein of claim 2, wherein the protein consists of a single polypeptide chain having a length of 157 201217528 150 to 160 amino acids. 15. The protein of claim 2, wherein the interleukin region is identical to the first IL-1 family of interleukins at at least 50 positions and is identical to the second il_i at at least 50 positions Family interleukin. 16. The protein of claim 2, wherein at least the residues 丨丨-丨 and 120-147 (according to the IL-1 β number) are collectively at least 9% identical to the corresponding residues on IL-IRa. 17. The protein of claim 3, wherein at least residues 1 to 6, 45 to 61 and 148 to 153 (according to the IL-1 β number) are collectively at least 90% identical to the corresponding residues on IL-Ιβ. base. 18. The protein of claim 3, wherein at least residues 1 to 8, 42 to 120, and 148 to 153 (according to the IL-Ιβ number) are collectively at least 90% identical to the corresponding residues on IL-Ιβ. base. 19. The protein of claim 3, wherein the interleukin region is at least 80°/. Same as Ρ01, Ρ02, Ρ〇3, Ρ04, or Ρ05. 20. The protein of claim 19, wherein the interleukin region is at least 90% identical to Ρ01, Ρ02, ρ〇3, Ρ04, or Ρ05. 21. The protein of claim 20, wherein the interleukin region is at least 95% identical to Ρ (Π, Ρ02, ρ〇3, Ρ04, or Ρ05. 22. The protein of claim 21 Wherein the interleukin region is the same as Ρ01, Ρ02 or Ρ03. 23. The protein of claim 21, wherein the interleukin region is the same as Ρ04. 24. The protein of claim 21 Wherein the interleukin region S 158 201217528 is identical to P05. 25. The protein of claim 2, wherein the interleukin region has higher thermal stability than IL-Ιβ or IL-IRa 26. The protein of claim 2, wherein the interleukin region has a Tm value that is at least 2 degrees higher than Il-ΐβ or IL-IRa. 27. A protein according to claim 3, wherein The protein has an IC50 of less than 10 ng/ml (ng/ml) for inhibition of IL-Ιβ (in ι·ng nanograms/ml (ng/ml)) in a cell signaling assay. 28. A protein as claimed in claim 3, wherein the protein is compared to human IL-I Ra is bound to IL-1RI with a higher affinity KD, a slower dissociation rate, and/or a faster rate of binding. 29. A protein according to claim 3, wherein the interleukin region Containing one or more of the following: (a) Site A: i. Site A residue, which is at least 80% identical to the corresponding residue on a first parenteral region; Π. a group, which is at least 80% identical to a corresponding residue on a first parent interleukin region; iii. A1 and A2 segments of site A, which are at least 80% identical to a first parental interleukin region Corresponding zone; and iv.-site A zone, which is at least 80% identical to the corresponding zone on a first parental interleukin region; (b) Site B ·· i. Site B residue, It is at least 80% identical to the corresponding residue on a 159 201217528 di-mother intercellular region; Π. the extended site B residue, which is at least 80% identical to a second parent interleukin region Corresponding residues; iii. B, B2 and B3 segments of site B, which are at least 80% identical to the corresponding regions on a second parenteral region a zone B; and a iv. zone B zone, which is at least 80% identical to a corresponding zone on a second parenteral region. 30. A pharmaceutical composition comprising, as claimed in claim 1 A protein according to any one of the preceding claims, wherein the protein comprises a protein according to any one of claims 1 to 29. 32. An ophthalmic pharmaceutical composition comprising as an application A protein according to any one of claims 1 to 29. 33. The pharmaceutical composition of claim 30, which is aqueous and comprises a protein having a concentration ranging from 0.001 to 5%. 34. Use of a composition as claimed in claim 30 for use in a body to modulate an immune or inflammatory response in a body. 35. Use of a composition as claimed in claim 30 for the treatment of an individual suffering from or at risk of a systemic autoimmune disease. 36. Use of a composition as claimed in claim 30 for the treatment of an individual suffering from or at risk of rheumatoid arthritis. 37. Use of a composition as claimed in claim 30 for the treatment of a dry eye disease or allergic conjunctivitis or at a risk thereof. 38. An isolated nucleic acid comprising a sequence encoding a protein according to any one of claims 1 to 29. 39. An isolated nucleic acid vector comprising a sequence encoding a protein according to any one of claims 1 to 29, wherein the sequence is operably linked to a transcriptional control sequence. 40. A recombinant host cell comprising a recombinant nucleic acid encoding a protein according to any one of claims 1 to 29. 41. A host cell as claimed in claim 42 which is an E. coli host cell. 42. A method for producing a recombinant protein, the method comprising: cultivating a host cell as claimed in claim 42 under conditions capable of expressing the recombinant protein; and purifying the recombinant protein from a cell lysate or a cell culture solution; And formulating the recombinant protein for administration to a body. 161