Target-tissue-specific antigen-binding molecule

Abstract

The present inventors discovered that the above-mentioned problems can be solved by producing antigen-binding molecules that contain an antigen-binding domain whose 5 antigen- binding activity varies depending on the concentration of a target tissue-specific compound. Use of antigen-binding molecules of the present invention enables various diseases that originate from a target tissue to be treated in a manner specific to the target tissue. 20 23 22 95 07 1 2 Se p 20 23 TARGET-TISSUE-SPECIFIC ANTIGEN-BINDING MOLECULE 12 Sep 2023 2023229507

Description

TARGET TISSUE-SPECIFIC ANTIGEN-BINDING MOLECULE Technical Field The present invention provides antigen-binding molecules comprising an antigenbinding domain whose antigen-binding activity varies depending on the concentration of a target tissue-specific compound; production methods and screening methods for the antigenbinding molecules; and pharmaceutical compositions containing the antigen-binding molecules. Cross-Reference to Related Applications This application is a divisional of Australian Patent Application No. 2020203710, which in turn is a divisional of Australian Patent Application No. 2018201358, which in turn is a divisional of Australian Patent Application No. 2013268418, the Australian National Phase application of International Application No. PCT / JP2013 / 064975, claiming priority to JP2012-123781 originally filed 30 May 2012 and JP2012-177311 originally filed 9 August 2012. The entire content of each of these applications is incorporated herein by reference. Reference to a Sequence Listing Preceding applications contained a Sequence Listing which was originally submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy is named "JPOXMLDOC01-seql.txt" and is 184,878 bytes in size. The present application contains a sequence listing which has been submitted electronically as an XML document in the ST.26 format and is hereby incorporated by reference in its entirety. Said XML copy, created on 7 September 2023, is named "P129446D3 Seq Listing.xml" and is 150,620 bytes in size. Background Art Antibodies are drawing attention as pharmaceuticals as they are highly stable in plasma and have few side effects. In particular, a number of IgG-type antibody pharmaceuticals are available on the market, and many antibody pharmaceuticals are currently under development (Non-Patent Documents 1 and 2). As cancer therapeutic agents using antibody pharmaceuticals, Rituxan against a CD20 antigen, cetuximab against an EGFR antigen, herceptin against a HER2 antigen, and such have been approved so far (Non-Patent Document 3). These antibody molecules bind to antigens expressed on cancer cells, and exhibit cytotoxic activity against cancer cells by ADCC and such. Such cytotoxic activity by ADCC and etc. are known to depend on the 2023229507  12 Sep 2023 number of antigens expressed on cells targeted by the therapeutic antibodies (Non-Patent Document 4); therefore, high expression level of the target antigen is preferable from the stand point of the effects of the therapeutic antibodies. However, even if the antigen expression level is high, when antigens are expressed in normal tissues, cytotoxic activity mediated by ADCC etc will be exerted against normal cells, and therefore side-effects will become a major problem. Therefore, antigens targeted by therapeutic antibodies used as therapeutic agents for cancer are preferably antigens specifically expressed in cancer cells. For example, antibody molecules against the EpCAM antigen which is known as a cancer antigen have been considered to be promising as therapeutic agents for cancer. However, the EpCAM antigen is known to be expressed in the pancreas as well, and in practice, administration of anti-EpCAM antibodies in clinical trials has been reported to cause pancreatitis as a side-effect due to cytotoxic activity towards the pancreas (Non-Patent Document 5). Following the success of antibody pharmaceuticals that exert cytotoxic activity by ADCC activity, a second generation of improved antibody molecules that exert strong cytotoxic activity through enhancement of ADCC activity by removing fucose of N-type sugar chains in 2023229507  12 Sep 2023 the native human IgG1 Fc region (Non-Patent Document 6), enhancement of ADCC activity by enhancing the binding toward FcYRIIIa by substitution of amino acids in the native human IgG1 Fc region (Non-Patent Document 7), and such have been reported. As antibody pharmaceuticals that exert cytotoxic activity against cancer cells through a mechanism other than 5 the above-mentioned ADCC activity mediated by NK cells, improved antibody molecules that exert a stronger cytotoxic activity, such as an antibody-drug conjugate (ADC) in which an antibody is conjugated with a drug having potent cytotoxic activity (Non-Patent Document 8), and a low molecular weight antibody that exerts toxic activity against cancer cells by recruiting T cells to cancer cells, have been reported as well. 0 Such antibody molecules exerting a stronger cytotoxic activity can exert cytotoxic activity against cancer cells that do not have much antigen expression, but on the other hand, they will exert similar cytotoxic activity against normal tissues with low antigen expression. In fact, in comparison to cetuximab which is a natural human IgG1 against an EGFR antigen, EGFR-BiTE, which is a bispecific antibody against CD3 and EGFR, can exert a potent cytotoxic 5 activity against cancer cells by recruiting T cells to cancer cells and exert antitumor effects. On the other hand, since EGFR is expressed also in normal tissues, when EGFR-BiTE is administered to cynomolgus monkeys, serious side effects have appeared (Non-Patent Document 10). Furthermore, bivatuzumab mertansine, an ADC formed by linking mertansine to an antibody against CD44v6 which is highly expressed in cancer cells, has been shown to cause 0 severe skin toxicity and liver toxicity in clinical practice because CD44v6 is expressed also in normal tissues (Non-Patent Document 11). When antibodies that can exert a potent cytotoxic activity against cancer cells having low antigen expression are used as such, the target antigen needs to be expressed in a highly cancer-specific manner. However, since HER2 and EGFR, which are target antigens of 25 herceptin and cetuximab, respectively, are also expressed in normal tissues, the number of cancer antigens expressed in a highly cancer-specific manner is thought to be limited. Therefore, while it is possible to strengthen the cytotoxic activity against cancer, the side effects occurring due to cytotoxic actions against normal tissues may become problematic. Furthermore, recently, ipilimumab which enhances tumor immunity by inhibiting 30 CTLA4 which contributes to immunosuppression in cancer was shown to prolong overall survival of metastatic melanoma (Non-Patent Document 12). However, since ipulimumab inhibits CTLA4 systemically, while tumor immunity is enhanced, the emergence of autoimmune disease-like severe side effects due to systemic activation of the immune system is becoming a problem (Non-Patent Document 13). 35 On the other hand, as antibody pharmaceuticals against diseases besides cancer, antibody pharmaceuticals that exert therapeutic effects by inhibiting inflammatory cytokines in 2023229507  12 Sep 2023 inflammatory / autoimmune diseases are known (Non-Patent Document 14). For example, Remicade and Humira which target TNF, and Actemra which targets IL-6R exhibit high therapeutic effects against rheumatoid arthritis, but on the other hand, systemic neutralization of these cytokines has led to the observation of infection as side effects (Non-Patent Document 15). 5 Various techniques have been developed as techniques that can be applied to second-generation antibody pharmaceuticals. While techniques for improving effector functions, antigen-binding ability, pharmacokinetics, and stability, or techniques for reducing immunogenic risks have been reported (Non-Patent Document 16), there are hardly any reports on techniques that enable target tissue-specific action of antibody pharmaceuticals to overcome 0 such side effects. For example, regarding lesions such as cancer tissues and inflammatory tissues, pH-dependent antibodies that make use of the acidic pH condition at these target tissues have been reported (Patent Documents 1 and 2). However, the decrease of pH (that is, increase in hydrogen ion concentration) in cancer tissues and inflammatory tissues as compared to normal tissues is slight, and since it is difficult to produce antibodies that act by detecting a slight 5 increase in the concentration of hydrogen ions which have an extremely small molecular weight, and also because acidic pH conditions may be found in normal tissues such as osteoclastic bone resorption region or in tissues other than the lesion of interest, use of pH conditions as a lesion-specific environmental factor was considered to face many challenges. On the other hand, methods for producing antibodies that exert antigen-binding activity only after they are 0 cleaved by a protease expressed at lesion sites such as cancer tissues and inflammatory tissues have been reported (Patent Document 3). However, since cleavage of antibodies by proteases is irreversible, when the antibodies that have been cleaved at the lesion site enter the blood stream and return to normal tissues, they can bind to the antigens in normal tissues as well, and this is considered to be a problem. Furthermore, cancer specificity of such proteases is also thought to 25 have problems that need to be addressed. Therefore, techniques that enable reversible action at sites of inflammation or cancer (lesion sites) without systemic action in normal tissues and blood for exerting drug efficacy while avoiding side effects are not known. [Prior art documents] 30 [Patent documents] [Patent document 1] WO 2003 / 105757 [Patent document 2] WO 2012 / 033953 [Patent document 3] WO 2010 / 081173 [Non-patent documents] 35 [Non-patent document 1] Monoclonal antibody successes in the clinic. Janice M Reichert, Clark J Rosensweig, Laura B Faden & Matthew C Dewitz, Nat. Biotechnol. (2005) 23, 1073 - 1078 2023229507  12 Sep 2023 [Non-patent document 2] The therapeutic antibodies market to 2008. Pavlou AK, Belsey MJ., Eur. J. Pharm. Biopharm. (2005) 59 (3), 389-396 [Non-patent document 3] Monoclonal antibodies: versatile platforms for cancer immunotherapy. Weiner LM, Surana R, Wang S., Nat. Rev. Immunol. (2010) 10 (5), 317-327 5 [Non-patent document 4] Differential responses of human tumor cell lines to anti-p185HER2 monoclonal antibodies. Lewis GD, Figari I, Fendly B, Wong WL, Carter P, Gorman C, Shepard HM, Cancer Immunol. Immunotherapy (1993) 37, 255-263 [Non-patent document 5] ING-1, a monoclonal antibody targeting Ep-CAM in patients with advanced adenocarcinomas. de Bono JS, Tolcher AW, Forero A, Vanhove GF, Takimoto C, Bauer 0 RJ, Hammond LA, Patnaik A, White ML, Shen S, Khazaeli MB, Rowinsky EK, LoBuglio AF, Clin. Cancer Res. (2004) 10 (22), 7555-7565 [Non-patent document 6] Non-fucosylated therapeutic antibodies as next-generation therapeutic antibodies. Satoh M, Iida S, Shitara K., Expert Opin. Biol. Ther. (2006) 6 (11), 1161-1173 [Non-patent document 7] Optimizing engagement of the immune system by anti-tumor 5 antibodies: an engineer's perspective. Desjarlais JR, Lazar GA, Zhukovsky EA, Chu SY., Drug Discov. Today (2007) 12 (21-22), 898-910 [Non-patent document 8] Antibody-drug conjugates: targeted drug delivery for cancer. Alley SC, Okeley NM, Senter PD., Curr. Opin. Chem. Biol. (2010) 14 (4), 529-537 [Non-patent document 9] BiTE: Teaching antibodies to engage T-cells for cancer therapy. 0 Baeuerle PA, Kufer P, Bargou R., Curr. Opin. Mol. Ther. (2009) 11 (1), 22-30 [Non-patent document 10] T cell-engaging BiTE antibodies specific for EGFR potently eliminate KRAS- and BRAF-mutated colorectal cancer cells. Lutterbuese R, Raum T, Kischel R, Hoffmann P, Mangold S, Rattel B, Friedrich M, Thomas O, Lorenczewski G, Rau D, Schaller E, Herrmann I, Wolf A, Urbig T, Baeuerle PA, Kufer P., Proc. Natl. Acad. Sci. U.S.A. (2010) 107 25 (28), 12605-12610 [Non-patent document 11] Phase I trial with the CD44v6-targeting immunoconjugate bivatuzumab mertansine in head and neck squamous cell carcinoma. Riechelmann H, Sauter A, Golze W, Hanft G, Schroen C, Hoermann K, Erhardt T, Gronau S., Oral Oncol. (2008) 44 (9), 823-829 30 [Non-patent document 12] Ipilimumab in the treatment of melanoma. Trinh VA, Hwu WJ., Expert Opin. Biol. Ther., (2012) Apr 14 (doi:10.1517 / 14712598.2012.675325) [Non-patent document 13] IPILIMUMAB - A NOVEL IMMUNOMODULATING THERAPY CAUSING AUTOIMMUNE HYPOPHYSITIS: A CASE REPORT AND REVIEW. Juszczak A, Gupta A, Karavitaki N, Middleton MR, Grossman A., Eur. J. Endocrinol. (2012) Apr 10 (doi: 35 10.1530 / EJE-12-0167) [Non-patent document 14] The Japanese experience with biologic therapies for rheumatoid 2023229507   29 Jun 2026 arthritis. Takeuchi T, Kameda H., Nat. Rev. Rheumatol. (2010) 6 (11), 644-652 [Non-patent document 15] Current evidence for the management of rheumatoid arthritis with biological disease-modifying antirheumatic drugs: a systematic literature review informing the EULAR recommendations for the management of RA. Nam JL, Winthrop KL, van Vollenhoven RF, Pavelka K, Valesini G, Hensor EM, Worthy G, Landewe R, Smolen JS, Emery P, Buch MH., Ann. Rheum. Dis. (2010) 69 (6), 976-986 [Non-patent document 16] Antibody engineering for the development of therapeutic antibodies. Kim SJ, Park Y, Hong HJ., Mol. Cells. (2005) 20 (1), 17-29 Any discussion of the prior art throughout the specification should not be considered as an express or implied admission that such prior art is widely known or forms part of the common general knowledge in the field. It is an object of the present invention to overcome or ameliorate one or more of the disadvantages of the prior art, or at least to provide a useful alternative. Summary of the Invention According to a first aspect, the present disclosure provides a method of production for an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a compound, the method comprising: (a) assaying binding of an antigen-binding domain to the antigen in the presence of a first concentration of a compound, wherein the compound is kynurenine, or a precursor or metabolite thereof; (b) assaying binding of the antigen-binding domain to the antigen in the presence of a second concentration of the compound that is different from the first concentration; (c) determining that the antigen-binding domain’s binding affinity for the antigen in the presence of the first concentration of the compound is different from the antigen-binding domain’s binding affinity for the antigen in the presence of the second concentration of the compound; and (d) selecting the antigen-binding domain based on the determination of (c), wherein either the first concentration or the second concentration, but not both, can be zero. According to a second aspect, the present disclosure provides a method of production for an antigen-binding domain whose antigen-binding activity varies depending on the concentration of 2023229507   29 Jun 2026 a compound, the method comprising: (a) contacting (i) an antigen with (ii) a plurality of different antigen-binding domains, in the presence of a first concentration of a compound, thereby forming one or more complexes, each comprising the antigen and an antigen-binding domain, wherein the compound is kynurenine, or a precursor or metabolite thereof; (b) exposing the one or more complexes of (a) to a second concentration of the compound different from the first concentration, thereby causing at least one of the complexes to dissociate into its constituent antigen and antigen-binding domain; and (c) selecting an antigen-binding domain that dissociated from the antigen in (b), wherein either the first concentration or the second concentration, but not both, can be zero. According to a third aspect, the present disclosure provides a method of production for an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a compound, the method comprising: (a) contacting (i) an antigen with (ii) a plurality of different antigen-binding domains, in the presence of a first concentration of a compound, wherein the compound is kynurenine, or a precursor or metabolite thereof; (b) isolating one or more antigen-binding domains that do not bind to the antigen in the presence of the first concentration of the compound; (c) assaying binding of the one or more isolated antigen-binding domains to the antigen in the presence of a second concentration of the compound, wherein the second concentration is different than the first concentration, and wherein either the first concentration or the second concentration, but not both, can be zero; and (d) selecting an antigen-binding domain that binds to the antigen in the presence of the second concentration of the compound. According to a fourth aspect, the present disclosure provides a method of screening for an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a compound, the method comprising: (a) assaying binding of an antigen-binding domain to the antigen in the presence of a first concentration of a compound, wherein the compound is kynurenine, or a precursor or metabolite thereof; 2023229507   29 Jun 2026 (b) assaying binding of the antigen-binding domain to the antigen in the presence of a second concentration of the compound that is different from the first concentration; (c) determining that the antigen-binding domain’s binding affinity for the antigen in the presence of the first concentration of the compound is different from the antigen-binding domain’s binding affinity for the antigen in the presence of the second concentration of the compound; and (d) selecting the antigen-binding domain based on the determination of (c), wherein either the first concentration or the second concentration, but not both, can be zero. According to a fifth aspect, the present disclosure provides a method of screening for an antigenbinding domain whose antigen-binding activity varies depending on the concentration of a compound, the method comprising: (a) contacting (i) an antigen with (ii) a plurality of different antigen-binding domains, in the presence of a first concentration of a compound, thereby forming one or more complexes, each comprising the antigen and an antigen-binding domain, wherein the compound is kynurenine, or a precursor or metabolite thereof; (b) exposing the one or more complexes of (a) to a second concentration of the compound different from the first concentration, thereby causing at least one of the complexes to dissociate into its constituent antigen and antigen-binding domain; and (c) selecting an antigen-binding domain that dissociated from the antigen in (b), wherein either the first concentration or the second concentration, but not both, can be zero. According to a sixth aspect, the present disclosure provides a method of screening for an antigenbinding domain whose antigen-binding activity varies depending on the concentration of a compound, the method comprising: (a) contacting (i) an antigen with (ii) a plurality of different antigen-binding domains, in the presence of a first concentration of a compound, wherein the compound is kynurenine, or a precursor or metabolite thereof; (b) isolating one or more antigen-binding domains that do not bind to the antigen in the presence of the first concentration of the compound; (c) assaying binding of the one or more isolated antigen-binding domains to the antigen in the presence of a second concentration of the compound, wherein the second concentration is different than the first concentration, and wherein either the first concentration or the second concentration, but not both, can be zero; and 2023229507   29 Jun 2026 (d) selecting an antigen-binding domain that binds to the antigen in the presence of the second concentration of the compound. In one aspect, the present invention provides pharmaceutical compositions that are useful for treating diseases originating from target tissues, and active ingredients thereof. In another aspect, the present invention provides methods of screening for the pharmaceutical compositions and active ingredients, as well as their production methods. The present inventors conducted dedicated studies. As a result, they generated antigen-binding molecules comprising an antigen-binding domain whose antigen-binding activity varies depending on the concentration of the target tissue-specific compound. Furthermore, the present inventors discovered that the antigen-binding molecules or pharmaceutical compositions comprising the antigen-binding molecules are useful for treating diseases that originate from a target tissue, and that they are also useful for treatment of diseases originating from target tissues that includes administering the antigen-binding molecules. They also discovered that the antigen-binding molecules are useful in the production of pharmaceuticals for treating diseases that originate from target tissues. Furthermore, the present inventors produced screening methods and production methods for the antigen-binding molecules, and thereby completed the present invention. More specifically, the present invention provides the following: [1] An antigen-binding molecule comprising an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a target tissue-specific compound. [2] The antigen-binding molecule of [1], wherein the target tissue is a cancer tissue. [3] The antigen-binding molecule of [2], wherein the compound specific to a cancer tissue is a metabolite specific to a cancer cell, a metabolite specific to an immune cell that has infiltrated 2023229507  12 Sep 2023 into a cancer tissue, or a metabolite specific to a stromal cell in a cancer tissue. [4] The antigen-binding molecule of [1], wherein the target tissue is an inflamed tissue. [5] The antigen-binding molecule of [4], wherein the compound specific to an inflamed tissue is a metabolite specific to an immune cell that has infiltrated into an inflamed tissue or a metabolite 5 specific to a normal cell that has been damaged in an inflamed tissue. [6] The antigen-binding molecule of [1], wherein the compound is at least one compound selected from a nucleoside having a purine ring structure, an amino acid and its metabolite, a lipid and its metabolite, a primary metabolite of glycometabolism, and nicotinamide and its metabolite. 0 [7] The antigen-binding molecule of [6], wherein the compound is at least one compound selected from adenosine, adenosine triphosphate, inosine, alanine, glutamic acid, aspartic acid, kynurenine, prostaglandin E2, succinic acid, citric acid, and 1-methylnicotinamide. [8] The antigen-binding molecule of any one of [1] to [7], wherein the antigen is a membrane-type molecule. 5 [9] The antigen-binding molecule of any one of [1] to [8], which is an antigen-binding molecule that has a neutralizing activity.

[10] The antigen-binding molecule of any one of [1] to [9], which is an antigen-binding molecule that has a cytotoxic activity.

[11] The antigen-binding molecule of any one of [1] to

[10] , which comprises an Fc region. 0

[12] The antigen-binding molecule of

[11] , wherein the Fc region is an Fc region contained in the constant region of SEQ ID NOs: 5, 6, 7, or 8.

[13] The antigen-binding molecule of

[11] , wherein the Fc region comprises an altered FcYR-binding Fc region that has a higher Fcy receptor-binding activity than the Fcy receptor-binding activity of a native human IgG Fc region. 25

[14] The antigen-binding molecule of

[13] , wherein at least one or more amino acids selected from the group consisting of amino acids at positions 221, 222, 223, 224, 225, 227, 228, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 246, 247, 249, 250, 251, 254, 255, 256, 258, 260, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 278, 279, 280, 281, 282, 283, 284, 285, 286, 288, 290, 291, 292, 293, 294, 295, 296, 297, 30 298, 299, 300, 301, 302, 303, 304, 305, 311, 313, 315, 317, 318, 320, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 339, 376, 377, 378, 379, 380, 382, 385, 392, 396, 421, 427, 428, 429, 434, 436, and 440 according to EU numbering, in the amino acid sequence of the altered FcYR-binding Fc region are different from the amino acids of the native human IgG Fc region. 35

[15] The antigen-binding molecule of

[14] , which comprises at least one or more amino acids selected from the group consisting of: 2023229507  12 Sep 2023 Lys or Tyr for the amino acid at position 221; Phe, Trp, Glu, or Tyr for the amino acid at position 222; Phe, Trp, Glu, or Lys for the amino acid at position 223; Phe, Trp, Glu, or Tyr for the amino acid at position 224; 5 Glu, Lys, or Trp for the amino acid at position 225; Glu, Gly, Lys, or Tyr for the amino acid at position 227; Glu, Gly, Lys, or Tyr for the amino acid at position 228; Ala, Glu, Gly, or Tyr for the amino acid at position 230; Glu, Gly, Lys, Pro, or Tyr for the amino acid at position 231; 0 Glu, Gly, Lys, or Tyr for the amino acid at position 232; Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 233; Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 234; 5 Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 235; Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 236; Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the 0 amino acid at position 237; Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 238; Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr for the amino acid at position 239; 25 Ala, Ile, Met, or Thr for the amino acid at position 240; Asp, Glu, Leu, Arg, Trp, or Tyr for the amino acid at position 241; Leu, Glu, Leu, Gln, Arg, Trp, or Tyr for the amino acid at position 243; His for the amino acid at position 244; Ala for the amino acid at position 245; 30 Asp, Glu, His, or Tyr for the amino acid at position 246; Ala, Phe, Gly, His, Ile, Leu, Met, Thr, Val, or Tyr for the amino acid at position 247; Glu, His, Gln, or Tyr for the amino acid at position 249; Glu or Gln for the amino acid at position 250; Phe for the amino acid at position 251; 35 Phe, Met, or Tyr for the amino acid at position 254; Glu, Leu, or Tyr for the amino acid at position 255; 2023229507  12 Sep 2023 Ala, Met, or Pro for the amino acid at position 256; Asp, Glu, His, Ser, or Tyr for the amino acid at position 258; Asp, Glu, His, or Tyr for the amino acid at position 260; Ala, Glu, Phe, Ile, or Thr for the amino acid at position 262; 5 Ala, Ile, Met, or Thr for the amino acid at position 263; Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Trp, or Tyr for the amino acid at position 264; Ala, Leu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 265; 0 Ala, Ile, Met, or Thr for the amino acid at position 266; Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr for the amino acid at position 267; Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Pro, Gln, Arg, Thr, Val, or Trp for the amino acid at position 268; 5 Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 269; Glu, Phe, Gly, His, Ile, Leu, Met, Pro, Gln, Arg, Ser, Thr, Trp, or Tyr for the amino acid at position 270; Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the 0 amino acid at position 271; Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 272; Phe or Ile for the amino acid at position 273; Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid 25 at position 274; Leu or Trp for the amino acid at position 275; Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 276; Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, or Trp for the amino 30 acid at position 278; Ala for the amino acid at position 279; Ala, Gly, His, Lys, Leu, Pro, Gln, Trp, or Tyr for the amino acid at position 280; Asp, Lys, Pro, or Tyr for the amino acid at position 281; Glu, Gly, Lys, Pro, or Tyr for the amino acid at position 282; 35 Ala, Gly, His, Ile, Lys, Leu, Met, Pro, Arg, or Tyr for the amino acid at position 283; Asp, Glu, Leu, Asn, Thr, or Tyr for the amino acid at position 284; 2023229507  12 Sep 2023 Asp, Glu, Lys, Gln, Trp, or Tyr for the amino acid at position 285; Glu, Gly, Pro, or Tyr for the amino acid at position 286; Asn, Asp, Glu, or Tyr for the amino acid at position 288; Asp, Gly, His, Leu, Asn, Ser, Thr, Trp, or Tyr for the amino acid at position 290; 5 Asp, Glu, Gly, His, Ile, Gln, or Thr for the amino acid at position 291; Ala, Asp, Glu, Pro, Thr, or Tyr for the amino acid at position 292; Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 293; Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at 0 position 294; Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 295; Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val for the amino acid at position 296; 5 Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 297; Ala, Asp, Glu, Phe, His, Ile, Lys, Met, Asn, Gln, Arg, Thr, Val, Trp, or Tyr for the amino acid at position 298; Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Val, Trp, or Tyr for the 0 amino acid at position 299; Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, or Trp for the amino acid at position 300; Asp, Glu, His, or Tyr for the amino acid at position 301; Ile for the amino acid at position 302; 25 Asp, Gly, or Tyr for the amino acid at position 303; Asp, His, Leu, Asn, or Thr for the amino acid at position 304; Glu, Ile, Thr, or Tyr for the amino acid at position 305; Ala, Asp, Asn, Thr, Val, or Tyr for the amino acid at position 311; Phe for the amino acid at position 313; 30 Leu for the amino acid at position 315; Glu, or Gln for the amino acid at position 317; His, Leu, Asn, Pro, Gln, Arg, Thr, Val, or Tyr for the amino acid at position 318; Asp, Phe, Gly, His, Ile, Leu, Asn, Pro, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 320; 35 Ala, Asp, Phe, Gly, His, Ile, Pro, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 322; Ile for the amino acid at position 323; 2023229507  12 Sep 2023 Asp, Phe, Gly, His, Ile, Leu, Met, Pro, Arg, Thr, Val, Trp, or Tyr for the amino acid at position 324; Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 325; 5 Ala, Asp, Glu, Gly, Ile, Leu, Met, Asn, Pro, Gln, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 326; Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg, Thr, Val, Trp, or Tyr for the amino acid at position 327; Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the 0 amino acid at position 328; Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 329; Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 330; 5 Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Thr, Val, Trp, or Tyr for the amino acid at position 331; Ala, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 332; Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Pro, Ser, Thr, Val, or Tyr for the amino acid at position 333; 0 Ala, Glu, Phe, Ile, Leu, Pro, or Thr for the amino acid at position 334; Asp, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Arg, Ser, Val, Trp, or Tyr for the amino acid at position 335; Glu, Lys, or Tyr for the amino acid at position 336; Glu, His, or Asn for the amino acid at position 337; 25 Asp, Phe, Gly, Ile, Lys, Met, Asn, Gln, Arg, Ser, or Thr for the amino acid at position 339; Ala or Val for the amino acid at position 376; Gly or Lys for the amino acid at position 377; Asp for the amino acid at position 378; Asn for the amino acid at position 379; 30 Ala, Asn, or Ser for the amino acid at position 380; Ala, or Ile for the amino acid at position 382; Glu for the amino acid at position 385; Thr for the amino acid at position 392; Leu for the amino acid at position 396; 35 Lys for the amino acid at position 421; Asn for the amino acid at position 427; 2023229507  12 Sep 2023 Phe, or Leu for the amino acid at position 428; Met for the amino acid at position 429; Trp for the amino acid at position 434; Ile for the amino acid at position 436; and 5 Gly, His, Ile, Leu, or Tyr for the amino acid at position 440 according to EU numbering in the amino acid sequence of the altered FcYR-binding Fc region.

[16] The antigen-binding molecule of

[11] , wherein the Fc region is modified so that there is a higher proportion of Fc region bound by a fucose-deficient sugar chain in a composition of sugar chain bound at position 297, according to EU numbering, of the Fc region, or so that there is a 0 higher proportion of Fc region with an added bisecting N-acetylglucosamine.

[17] The antigen-binding molecule of any one of

[11] and

[13] to

[16] , wherein the FcRn-binding activity of the Fc region under an acidic pH range condition is enhanced compared to the FcRn-binding activity of the Fc region of SEQ ID NO: 5, 6, 7, or 8.

[18] The antigen-binding molecule of

[17] , wherein the Fc region is an Fc region with 5 substitution of at least one or more amino acids selected from the group consisting of amino acids at positions 238, 244, 245, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 260, 262, 265, 270, 272, 279, 283, 285, 286, 288, 293, 303, 305, 307, 308, 309, 311, 312, 314, 316, 317, 318, 332, 339, 340, 341, 343, 356, 360, 362, 375, 376, 377, 378, 380, 382, 385, 386, 387, 388, 389, 400, 413, 415, 423, 424, 427, 428, 430, 431, 433, 434, 435, 436, 438, 439, 440, 442, and 447, 0 according to EU numbering, in the amino acid sequence of the Fc region comprised in the constant region of SEQ ID NO: 5, 6, 7, or 8.

[19] The antigen-binding molecule of

[18] , wherein the Fc region comprises at least one or more amino acids selected from the group consisting of: Leu for the amino acid at position 238; 25 Leu for the amino acid at position 244; Arg for the amino acid at position 245; Pro for the amino acid at position 249; Gln or Glu for the amino acid at position 250; Arg, Asp, Glu, or Leu for the amino acid at position 251; 30 Phe, Ser, Thr, or Tyr for the amino acid at position 252; Ser or Thr for the amino acid at position 254; Arg, Gly, Ile, or Leu for the amino acid at position 255; Ala, Arg, Asn, Asp, Gln, Glu, Pro, or Thr for the amino acid at position 256; Ala, Ile, Met, Asn, Ser, or Val for the amino acid at position 257; 35 Asp for the amino acid at position 258; Ser for the amino acid at position 260; 2023229507  12 Sep 2023 Leu for the amino acid at position 262; Lys for the amino acid at position 270; Leu, or Arg for the amino acid at position 272; Ala, Asp, Gly, His, Met, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr for the amino acid at position 279; 5 Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Ser, Thr, Trp, or Tyr for the amino acid at position 283; Asn for the amino acid at position 285; Phe for the amino acid at position 286; Asn or Pro for the amino acid at position 288; 0 Val for the amino acid at position 293, Ala, Glu, Gln, or Met for the amino acid at position 307; Ala, Glu, Ile, Lys, Leu, Met, Ser, Val, or Trp for the amino acid at position 311; Pro for the amino acid at position 309; Ala, Asp, or Pro for the amino acid at position 312; 5 Ala or Leu for the amino acid at position 314; Lys for the amino acid at position 316; Pro for the amino acid at position 317; Asn or Thr for the amino acid at position 318; Phe, His, Lys, Leu, Met, Arg, Ser, or Trp for the amino acid at position 332; 0 Asn, Thr, or Trp for the amino acid at position 339; Pro for the amino acid at position 341; Glu, His, Lys, Gln, Arg, Thr, or Tyr for the amino acid at position 343; Arg for the amino acid at position 375; Gly, Ile, Met, Pro, Thr, or Val for the amino acid at position 376; 25 Lys for the amino acid at position 377; Asp, Asn, or Val for the amino acid at position 378; Ala, Asn, Ser, or Thr for the amino acid at position 380; Phe, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 382; 30 Ala, Arg, Asp, Gly, His, Lys, Ser, or Thr for the amino acid at position 385; Arg, Asp, Ile, Lys, Met, Pro, Ser, or Thr for the amino acid at position 386; Ala, Arg, His, Pro, Ser, or Thr for the amino acid at position 387; Asn, Pro, or Ser for the amino acid at position 389; Asn for the amino acid at position 423; 35 Asn for the amino acid at position 427; Leu, Met, Phe, Ser, or Thr for the amino acid at position 428; 2023229507  12 Sep 2023 Ala, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, or Tyr for the amino acid at position 430; His or Asn for the amino acid at position 431; Arg, Gln, His, Ile, Lys, Pro, or Ser for the amino acid at position 433; 5 Ala, Gly, His, Phe, Ser, Trp, or Tyr for the amino acid at position 434; Arg, Asn, His, Ile, Leu, Lys, Met, or Thr for the amino acid at position 436; Lys, Leu, Thr, or Trp for the amino acid at position 438; Lys for the amino acid at position 440; Lys for the amino acid at position 442; and 0 Ile, Pro, or Thr for the amino acid at position 308; as indicated by EU numbering, in the amino acid sequence of the Fc region comprised in the constant region of SEQ ID NO: 5, 6, 7, or 8.

[20] The antigen-binding molecule of any one of [1] to

[19] , wherein the antigen-binding domain is a multispecific or a multiparatopic antigen-binding domain. 5

[21] The antigen-binding molecule of

[20] , wherein an antigen bound by at least one of the antigen-binding domains is a membrane-type molecule expressed on a cancer cell membrane, and an antigen bound by at least one of the antigen-binding domains is a membrane-type molecule expressed on an effector cell membrane.

[22] The antigen-binding molecule of

[21] , wherein the effector cell is an NK cell, a macrophage, 0 or a T cell.

[23] The antigen-binding molecule of

[21] or

[22] , wherein the membrane-type molecule expressed on an effector cell membrane is a TCR-constituting polypeptide, CD2, CD3, CD28, CD44, CD16, CD32, CD64, or NKG2D.

[24] The antigen-binding molecule of

[20] , wherein an antigen bound by at least one of the 25 antigen-binding domains is a membrane-type molecule expressed on a cancer cell membrane, and an antigen bound by at least one of the antigen-binding domains is a cytotoxic substance.

[25] The antigen-binding molecule of any one of

[20] to

[24] , wherein the antigen-binding molecule is an antibody fragment.

[26] The antigen-binding molecule of any one of [1] to

[24] , wherein the antigen-binding 30 molecule is an antibody.

[27] The antigen-binding molecule of any one of [1] to [7], wherein the antigen is a soluble molecule.

[28] The antigen-binding molecule of

[27] , which is an antigen-binding molecule having a neutralizing activity. 35

[29] The antigen-binding molecule of

[27] or

[28] , which comprises an Fc region.

[30] The antigen-binding molecule of

[29] , wherein the Fc region is an Fc region comprised in 2023229507  12 Sep 2023 the constant region of SEQ ID NO: 5, 6, 7, or 8.

[31] The antigen-binding molecule of

[29] , wherein the FcRn-binding activity of the Fc region under an acidic pH range condition is enhanced compared to the FcRn-binding activity of the Fc region comprised in the constant region of SEQ ID NO: 5, 6, 7, or 8. 5

[32] The antigen-binding molecule of

[31] , wherein the Fc region is an Fc region with substitution of at least one or more amino acids selected from the group consisting of amino acids at positions 238, 244, 245, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 260, 262, 265, 270, 272, 279, 283, 285, 286, 288, 293, 303, 305, 307, 308, 309, 311, 312, 314, 316, 317, 318, 332, 339, 340, 341, 343, 356, 360, 362, 375, 376, 377, 378, 380, 382, 385, 386, 387, 388, 389, 0 400, 413, 415, 423, 424, 427, 428, 430, 431, 433, 434, 435, 436, 438, 439, 440, 442, and 447, according to EU numbering, in the amino acid sequence of the Fc region comprised in the constant region of SEQ ID NO: 5, 6, 7, or 8.

[33] The antigen-binding molecule of

[32] , wherein the Fc region comprises at least one or more amino acids selected from the group consisting of: 5 Leu for the amino acid at position 238; Leu for the amino acid at position 244; Arg for the amino acid at position 245; Pro for the amino acid at position 249; Gln or Glu for the amino acid at position 250; 0 Arg, Asp, Glu, or Leu for the amino acid at position 251; Phe, Ser, Thr, or Tyr for the amino acid at position 252; Ser or Thr for the amino acid at position 254; Arg, Gly, Ile, or Leu for the amino acid at position 255; Ala, Arg, Asn, Asp, Gln, Glu, Pro, or Thr for the amino acid at position 256; 25 Ala, Ile, Met, Asn, Ser, or Val for the amino acid at position 257; Asp for the amino acid at position 258; Ser for the amino acid at position 260; Leu for the amino acid at position 262; Lys for the amino acid at position 270; 30 Leu,or Arg for the amino acid at position 272; Ala, Asp, Gly, His, Met, Asn, Gln, Arg, Ser, Thr, Trp, or Tyr for the amino acid at position 279; Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Arg, Ser, Thr, Trp, or Tyr for the amino acid at position 283; Asn for the amino acid at position 285; 35 Phe for the amino acid at position 286; Asn or Pro for the amino acid at position 288; 2023229507  12 Sep 2023 Val for the amino acid at position 293, Ala, Glu, Gln, or Met for the amino acid at position 307; Ala, Glu, Ile, Lys, Leu, Met, Ser, Val, or Trp for the amino acid at position 311; Pro for the amino acid at position 309; 5 Ala, Asp, or Pro for the amino acid at position 312; Ala or Leu for the amino acid at position 314; Lys for the amino acid at position 316; Pro for the amino acid at position 317; Asn or Thr for the amino acid at position 318; 0 Phe, His, Lys, Leu, Met, Arg, Ser, or Trp for the amino acid at position 332; Asn, Thr, or Trp for the amino acid at position 339; Pro for the amino acid at position 341; Glu, His, Lys, Gln, Arg, Thr, or Tyr for the amino acid at position 343; Arg for the amino acid at position 375; 5 Gly, Ile, Met, Pro, Thr, or Val for the amino acid at position 376; Lys for the amino acid at position 377; Asp, Asn, or Val for the amino acid at position 378; Ala, Asn, Ser, or Thr for the amino acid at position 380; Phe, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid at 0 position 382; Ala, Arg, Asp, Gly, His, Lys, Ser, or Thr for the amino acid at position 385; Arg, Asp, Ile, Lys, Met, Pro, Ser, or Thr for the amino acid at position 386; Ala, Arg, His, Pro, Ser, or Thr for the amino acid at position 387; Asn, Pro, or Ser for the amino acid at position 389; 25 Asn for the amino acid at position 423; Asn for the amino acid at position 427; Leu, Met, Phe, Ser, or Thr for the amino acid at position 428; Ala, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, or Tyr for the amino acid at position 430; 30 His or Asn for the amino acid at position 431; Arg, Gln, His, Ile, Lys, Pro, or Ser for the amino acid at position 433; Ala, Gly, His, Phe, Ser, Trp, or Tyr for the amino acid at position 434; Arg, Asn, His, Ile, Leu, Lys, Met, or Thr for the amino acid at position 436; Lys, Leu, Thr, or Trp for the amino acid at position 438; 35 Lys for the amino acid at position 440; Lys for the amino acid at position 442; and 2023229507  12 Sep 2023 Ile, Pro, or Thr for the amino acid at position 308; as indicated by EU numbering, in the amino acid sequence of the Fc region comprised in the constant region of SEQ ID NO: 5, 6, 7, or 8.

[34] The antigen-binding molecule of

[29] , wherein the FcRn-binding activity of the Fc region 5 under a neutral pH range condition is enhanced compared to the FcRn-binding activity of the Fc region comprised in the constant region of SEQ ID NO: 5, 6, 7, or 8.

[35] The antigen-binding molecule of

[34] , wherein the Fc region is an Fc region with substitution of at least one or more amino acids selected from the group consisting of amino acids at positions 237, 248, 250, 252, 254, 255, 256, 257, 258, 265, 286, 289, 297, 298, 303, 305, 0 307, 308, 309, 311, 312, 314, 315, 317, 332, 334, 360, 376, 380, 382, 384, 385, 386, 387, 389, 424, 428, 433, 434, and 436 according to EU numbering, in the amino acid sequence of the Fc region comprised in the constant region of SEQ ID NO: 5, 6, 7, or 8.

[36] The antigen-binding molecule of

[35] , wherein the Fc region comprises at least one or more amino acids selected from the group consisting of: 5 Met for the amino acid at position 237; Ile for the amino acid at position 248; Ala, Phe, Ile, Met, Gln, Ser, Val, Trp, or Tyr for the amino acid at position 250; Phe, Trp, or Tyr for the amino acid at position 252; Thr for the amino acid at position 254; 0 Glu for the amino acid at position 255; Asp, Asn, Glu, or Gln for the amino acid at position 256; Ala, Gly, Ile, Leu, Met, Asn, Ser, Thr, or Val for the amino acid at position 257; His for the amino acid at position 258; Ala for the amino acid at position 265; 25 Ala or Glu for the amino acid at position 286; His for the amino acid at position 289; Ala for the amino acid at position 297; Ala for the amino acid at position 303; Ala for the amino acid at position 305; 30 Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Val, Trp, or Tyr for the amino acid at position 307; Ala, Phe, Ile, Leu, Met, Pro, Gln, or Thr for the amino acid at position 308; Ala, Asp, Glu, Pro, or Arg for the amino acid at position 309; Ala, His, or Ile for the amino acid at position 311; 35 Ala or His for the amino acid at position 312; Lys or Arg for the amino acid at position 314; 2023229507  12 Sep 2023 Ala, Asp, or His for the amino acid at position 315; Ala for the amino acid at position 317; Val for the amino acid at position 332; Leu for the amino acid at position 334; 5 His for the amino acid at position 360; Ala for the amino acid at position 376; Ala for the amino acid at position 380; Ala for the amino acid at position 382; Ala for the amino acid at position 384; 0 Asp or His for the amino acid at position 385; Pro for the amino acid at position 386; Glu for the amino acid at position 387; Ala or Ser for the amino acid at position 389; Ala for the amino acid at position 424; 5 Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Ser, Thr, Val, Trp, or Tyr for the amino acid at position 428; Lys for the amino acid at position 433; Ala, Phe, His, Ser, Trp, or Tyr for the amino acid at position 434; and His, Ile, Leu, Phe, Thr, or Val for the amino acid at position 436 0 as indicated by EU numbering, in the amino acid sequence of the Fc region of SEQ ID NO: 5, 6, 7, or 8.

[37] The antigen-binding molecule of any one of

[29] and

[31] to

[36] , wherein the Fc region has a higher binding activity to an inhibitory Fcy receptor than to an activating Fcy receptor.

[38] The antigen-binding molecule of

[37] , wherein the inhibitory Fcy receptor is human 25   FcyRIIb.

[39] The antigen-binding molecule of

[37] or

[38] , wherein the activating Fcy receptor is human FcyRIa, human FcyRIIa (R), human FcyRIIa (H), human FcyRIIIa (V), or human FcyRIIIa (F).

[40] The antigen-binding molecule of any one of

[37] to

[39] , wherein the amino acid at position 238 or 328 (EU numbering) of the Fc region includes an amino acid that is different from the 30 amino acid of the native human IgG Fc region.

[41] The antigen-binding molecule of

[40] , wherein the amino acid at position 238 indicated by EU numbering in the Fc region is Asp or the amino acid at position 328 is Glu.

[42] The antigen-binding molecule of

[40] or

[41] , which comprises at least one or more amino acids selected from the group consisting of: 35 Asp for the amino acid at position 233; Trp or Tyr for the amino acid at position 234; 2023229507  12 Sep 2023 Ala, Asp, Glu, Leu, Met, Phe, Trp, or Tyr for the amino acid at position 237; Asp for the amino acid at position 239; Ala, Gln, or Val for the amino acid at position 267; Asn, Asp, or Glu for the amino acid at position 268; 5 Gly for the amino acid at position 271; Ala, Asn, Asp, Gln, Glu, Leu, Met, Ser, or Thr for the amino acid at position 326; Arg, Lys, or Met for the amino acid at position 330; Ile, Leu, or Met for the amino acid at position 323; and Asp for the amino acid at position 296 according to EU numbering, in the amino acid sequence 0 of the Fc region.

[43] The antigen-binding molecule of any one of

[27] to

[42] , wherein the antigen-binding molecule is an antibody.

[44] A method for producing the antigen-binding molecule of any one of [1] to

[43] , which comprises selecting an antigen-binding domain whose antigen-binding activity varies depending 5 on the concentration of a target tissue-specific compound.

[45] A method of screening for the antigen-binding molecule of any one of [1] to

[43] , which comprises selecting an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a target tissue-specific compound.

[46] A pharmaceutical composition comprising the antigen-binding molecule of any one of [1] to 0

[43] . Brief Description of the Drawings Fig. 1 shows that a small-molecule-switch antibody does not bind to antigens in a normal environment where the small molecules are not present, but binds to the antigens in the 25 target tissue where the small molecules are present at a high concentration. Fig. 2 shows that the small molecule functions as a switch by fitting between the anti-small-molecule antibody and the antigen. If the small molecule is absent, the antibody-antigen interaction is insufficient and the antibody cannot bind to the antigen, but if the small molecule is present, the antibody can bind to the antigen by having the small molecule 30 placed between the antibody and the antigen. Fig. 3 is a figure showing the result of ELISA for the binding of the antibody to human IL-6. The vertical axis shows the absorbance values which assess the binding activity to human IL-6 of each of the antibodies in the presence or absence of each of the small molecules. Fig. 4 is a sensorgram showing the interaction between 4 umolf of human IL-6 and 35 A11 in the presence or absence of 100 umolL kynurenine. Fig. 5 shows a graph that evaluates change in the response of binding to A11 2023229507  12 Sep 2023 immobilized onto Sensorchip CM5, when interaction is allowed to take place for 60 seconds with 1 pmol / L of IL-6 as the analyte. The vertical axis shows change in the response (RU) before and after IL-6 interaction, and the horizontal axis shows the concentration of kynurenine (pmol / L) contained in the solution at that time. 5 Fig. 6 shows a graph that evaluates the response to H01 which has been immobilized onto Sensorchip CM5, when interaction is allowed to take place for 60 seconds with 1 pmol / L of IL-6 as the analyte. The vertical axis shows change in the response (RU) before and after IL-6 interaction, and the horizontal axis shows the concentration of kynurenine contained in the solution (pmol / L) at that time. 0 Fig. 7 shows a graph that evaluates the response to IL-6 which has been immobilized onto Sensorchip CM5, when interaction is allowed to take place for 60 seconds with 0.1 pmol / L of A11 as the analyte. The vertical axis shows change in the response (RU) before and after A11 interaction, and the horizontal axis shows the concentration of kynurenine contained in the solution (pmol / L). 5 Fig. 8 shows a graph obtained by allowing A11 to interact with IL-6 immobilized on Sensorchip CM5 in the presence of 100 pmol / L kynurenine, and then observing the dissociation of A11 from IL6 in the presence of a buffer containing 100 pmol / L kynurenine or in the presence of a buffer that does not contain kynurenine. In the figure, the vertical axis shows values normalized by defining the amount of A11 bound in the presence of 100 pmol / L kynurenine as 0 100, and the horizontal axis shows the passage of time (in seconds) from the start of the interaction. Fig. 9 shows a sensorgram obtained by allowing 800, 400, 200, 100, 50, or 25 nmol / L of kynurenine to interact with IL-6 immobilized on a sensorchip. The vertical axis shows change in the amount of IL-6 bound by kynurenine (RU) (the response at the start of the interaction 25 experiment was defined as 0), and the horizontal axis shows the passage of time from the start of the interaction experiment. Fig. 10 shows the structure of 2'-Adenosine-PEG-peptide which is an adenosine analog used for immunization of rabbits. Fig. 11 shows the structure of 5'-Adenosine-PEG-peptide which is an adenosine analog 30 used for immunization of rabbits. Fig. 12 shows the structure of 2'-Adenosine-PEG-biotin produced by substituting biotin for the peptide portion of the adenosine analog used for immunization of rabbits. Fig. 13 shows the structure of 5'-Adenosine-PEG-biotin produced by substituting biotin for the peptide portion of the adenosine analog used for immunization of rabbits. 35 Fig. 14 is a graph where the vertical axis shows the value (N_binding_100) obtained by dividing the amount of binding in the interaction of each antibody with 2'-Adenosine-PEG-biotin 2023229507  12 Sep 2023 by the capture level (RU) of each antibody, and the horizontal axis shows the value (N_stability_100) obtained by dividing the value obtained 60 seconds after dissociation of 2'-Adenosine-PEG-biotin from each antibody after interaction with 2'-Adenosine-PEG-biotin by the capture level (RU) of each antibody. 5 Fig. 15A indicates sensorgrams of surface plasmon resonance-based analyses which show that clone SMB0002 binds to (interacts with) adenosine. The sensorgrams show the interactions between SMB0002 and the antigen at 7.81, 31.3, 125, and 500 nM in order from the bottom. Fig. 15B indicates sensorgrams of surface plasmon resonance-based analyses which 0 show that clone SMB0002 binds to (interacts with) ATP. The sensorgrams show the interactions between SMB0002 and the antigen at 78.1, 313, 1250, and 5000 nM in order from the bottom. Fig. 15C indicates sensorgrams of surface plasmon resonance-based analyses which show that clone SMB0089 binds to (interacts with) adenosine. The sensorgrams show the 5 interactions between SMB0089 and the antigen at 7.81, 31.3, 125, and 500 nM in order from the bottom. Fig. 15D indicates sensorgrams of surface plasmon resonance-based analyses which show that clone SMB0089 binds to (interacts with) ATP. The sensorgrams show the interactions between SMB00089 and the antigen at 78.1, 313, 1250, and 5000 nM in order from 0 the bottom. Fig. 15E indicates sensorgrams of surface plasmon resonance-based analyses which show that clone SMB0104 binds to (interacts with) adenosine. The sensorgrams show the interactions between SMB0104 and the antigen at 7.81, 31.3, and 500 nM in order from the bottom. 25 Fig. 15F indicates sensorgrams of surface plasmon resonance-based analyses which show that clone SMB0104 binds to (interacts with) ATP. The sensorgrams show the interactions between SMB0104 and the antigen at 78.1, 313, 1250, and 5000 nM in order from the bottom. Fig. 16 indicates sensorgrams of surface plasmon resonance-based analyses which show 30 that clone SMB0171 binds to (interacts with) ATP. The sensorgrams show the interactions between SMB0171 and the antigen at 5 and 50 uM in order from the bottom. Fig. 17 indicates the results of competitive ELISA which shows that clone SMB0002 binds to adenosine and ATP. Fig. 18 shows a graph that assesses the inhibitive ability of ATP towards binding of 35 biotin-labeled antigens (a mixture of 5'-Adenosine-PEG-biotin and ATP-PEG-biotin) by ATNLSA1-4_D12. 2023229507  12 Sep 2023 Fig. 19 shows the concept of a rationally designed antibody library that can obtain adenosine / ATP-switch antibodies against any antigen, wherein the library is made from antibody variable region portions that contact with the antigens such that adenosine or ATP is positioned between the antibody and antigen. 5 Fig. 20 shows the concept of an adenosine-immunized rabbit antibody library which yields adenosine / ATP-switch antibodies against any antigen and in which adenosine or ATP is sandwiched between the antibody and the antigen. Fig. 21 is a figure showing the result of ELISA for binding of the antibody to human IL-6. The vertical axis shows the binding activity of each antibody to human IL-6 depending 0 on the presence or absence of amino acids or amino acid metabolites (kynurenine, tryptophan, phenylalanine, anthranilic acid, 3-hydroxykynurenine, and kynurenic acid), presented as absorbance values at wavelength of 450 nm. Fig. 22 is a figure showing the result of ELISA for binding of the antibody to human IL-6. The vertical axis shows the binding activity of the I6NMSC1-3_#03 antibody to human 5 IL-6 depending on the presence or absence of each small molecule (ATP, adenosine, inosine, PGE2, succinic acid, lactic acid, kynurenine, and a small-molecule cocktail), presented as specific activity values calculated from absorbance values at wavelength of 450 nm. Fig. 23 is a figure showing the result of ELISA for binding of the antibody to human IL-6. The vertical axis shows the binding activity of the I6NMSC1-3_#17 antibody to human 0 IL-6 depending on the presence or absence of each small molecule (ATP, adenosine, inosine, PGE2, succinic acid, lactic acid, kynurenine, and a small-molecule cocktail), presented as specific activity values calculated from absorbance values at wavelength of 450 nm. Fig. 24 is a figure showing the result of ELISA for binding of the antibody to HSA. The vertical axis shows the binding activity of the HSNMSC1-4_#22 antibody to HSA 25 depending on the presence or absence of each small molecule (ATP, adenosine, inosine, PGE2, succinic acid, lactic acid, kynurenine, and a small-molecule cocktail), presented as absorbance values at wavelength of 450 nm. Fig. 25 is a figure showing the result of ELISA performed on clone I6DL2C5-4_076, which was obtained from the rationally designed antibody library against human IL-6 in the 30 presence or absence of ATP and / or adenosine at 1 mM. The vertical axis shows the absorbance value which evaluates binding activity of the antibody to human IL-6. Results obtained when using M13KO7 Helper Phage are presented as the negative control. Fig. 26 is a figure showing the result of ELISA performed on clone HSDL3C5-4_015, which was obtained from the rationally designed antibody library against human serum albumin 35 in the presence or absence of ATP and / or adenosine at 1 mM. The vertical axis shows the absorbance value which assesses binding activity of the antibody to human serum albumin. 2023229507  12 Sep 2023 Results obtained when using M13KO7 Helper Phage are presented as the negative control. Fig. 27 is a figure showing the result of ELISA performed on clone 6RAD2C1-4_011 and 6RAD2C1-4_076, which were obtained from the rationally designed antibody library against human IL-6 receptor in the presence or absence of ATP and / or adenosine (written as ADO) at 1 5 mM, and in the presence or absence of a small-molecule cocktail (SC). The vertical axis shows absorbance values which assess the binding activity of the antibody to the human IL-6 receptor. Results obtained when using M13KO7 Helper Phage are presented as the negative control. Fig. 28 is a figure showing the result of ELISA for binding of clone 6RNMSC1-2_F02 to human IL-6R. The vertical axis shows the absorbance values which assess the binding 0 activity of the antibody to human IL-6R in the presence or absence of each small molecule. Fig. 29 is a figure showing the result of ELISA for binding of clone 6RNMSC1-3_G02 to human IL-6R. The vertical axis shows the absorbance values which assess the binding activity of the antibody to human IL-6R in the presence or absence of each small molecule. Fig. 30 is a figure showing the result of ELISA for binding of an antibody to human 5 IL-6R. The vertical axis shows the absorbance values which assess the binding activity of the antibody to human IL-6R in the presence or absence of each amino acid or amino acid metabolite. Fig. 31 presents sensorgrams showing the interaction between 6RNMSC1-2_F02 and 1 pmol / L IL-6R in the presence of 100 pmolf kynurenine, in the presence of 10 mmol / L ATP, and 0 in the absence of kynurenine and ATP. The solid line indicates the interaction in the presence of kynurenine, the dotted line indicates the interaction in the presence of ATP, and the dashed line indicates the interaction in their absence. Fig. 32 is a graph obtained by allowing 6RNMSC1-2_F02 to interact with IL-6R immobilized on Sensorchip CM5 in the presence of 100 pmol / L kynurenine, and then observing 25 the dissociation of 6RNMSC1-2_F02 from IL-6R in the presence of a buffer containing 100 pmol / L kynurenine or in the presence of a buffer that does not contain kynurenine. In the figure, the vertical axis shows values normalized by defining the amount of 6RNMSC1-2_F02 bound in the presence of 100 pmol / L kynurenine as 100, and the horizontal axis shows the passage of time (in seconds) from the start of the interaction. The solid line shows the 30 dissociation of 6RNMSC1-2_F02 from IL-6R in the presence of kynurenine, and the dotted line shows the dissociation of 6RNMSC1-2_F02 from IL-6R in the absence of kynurenine. Fig. 33 is a graph produced by allowing 5 pg / L of 6RNMSC1-2_F02 to interact as an analyte for 180 seconds, and assessing the response to IL-6R immobilized onto Sensorchip CM5. The vertical axis shows change in the response (RU) before and after 6RNMSC1-2_F02 35 interaction, and the horizontal axis shows the concentration (pmol / L) of kynurenine contained in the solution. 2023229507  12 Sep 2023 Fig. 34 is a figure showing assessment of the binding of antibodies to membrane-type human IL-6R by FCM. The top panel shows results obtained in the presence of Kynurenine, and the bottom panel shows results obtained in the absence of Kynurenine. The horizontal axis shows the fluorescence intensity and the vertical axis shows the cell count. 5 Fig. 35A shows the ADCC activity of antibodies that bind to antigens in the presence of small molecules toward cells expressing the antigens. It shows the ADCC activity of clone 6RNMSC1-2_F02, which binds to hIL-6R in the presence of kynurenine, toward BaF cells expressing hIL-6R in the presence (triangles) or absence (circles) of kynurenine. The open triangles and circles show the measured values, and the filled triangles and circles show the 0 mean values. Fig. 35B shows the ADCC activity of antibodies that bind to antigens in the presence of small molecules toward cells that express the antigen. It shows the ADCC activity of MRA, which binds to hIL-6R regardless of the presence of kynurenine, toward BaF cells expressing hIL-6R in the presence (triangles) or absence (circles) of kynurenine. The open triangles and 5 circles show the measured values, and the filled triangles and circles show the mean values. Fig. 36 shows the ADCC activity of antibodies that bind to antigens in the presence of small molecules toward cells expressing the antigen. It shows the ADCC activity of clone 6RNMSC1-2_F02 toward BaF cells expressing hIL-6R in the presence (triangles) or absence (circles) of clone 6RNMSC1-2_F02 which binds to hIL-6R in the presence of kynurenine. The 0 horizontal axis shows the kynurenine concentration and the vertical axis shows the ADCC activity (%). The mean values and standard deviations of ADCC activity are shown. Fig. 37 is a figure showing the result of ELISA for the binding of clone 6RNMSC1-2_F02 in mouse serum to human IL-6R. The vertical axis shows the absorbance values which evaluate the binding activities of the antibody to human IL-6R in the presence or 25 absence of kynurenine. Fig. 38 is a figure showing the result of ELISA performed with clone I6RLSA1-6_011, which was obtained from the rationally designed antibody library, against human IL-6 in the presence or absence of ATP and adenosine at 10 mM. The vertical axis shows the absorbance value which evaluates binding activity of the antibody to human IL-6. Results obtained when 30 using a clone obtained from the rationally designed antibody library and showing binding activity toward human IL-6 regardless of the presence of small molecules are presented as the positive control. Results obtained when using the M13KO7 Helper Phage are presented as the negative control. Fig. 39 is a figure showing the result of ELISA performed with clone 6RRLSA1-6_037 35 and 6RRLSA1-6_045, which were obtained from the rationally designed antibody library, against the human IL-6 receptor in the presence or absence of ATP and adenosine at 10 mM. 2023229507  12 Sep 2023 The vertical axis shows the absorbance value which evaluates the binding activity of the antibodies to the human IL-6 receptor. Results obtained when using the M13KO7 Helper Phage are presented as the negative control. Fig. 40 is a figure showing the result of ELISA performed on 96 clones obtained by 5 panning the rationally designed antibody library four times against human IgA-Fc using a multivalent antibody phage display. The absorbance values which evaluate the binding activity of the antibodies to human IgA-Fc in the absence of ATP and adenosine are shown on the vertical axis, and absorbance values which evaluate the binding activity of the antibodies to human IgA-Fc in the presence of ATP and adenosine are shown on the horizontal axis. 0 Fig. 41 is a figure showing the result of ELISA performed on 96 clones obtained by panning the rationally designed antibody library four times against human IgA-Fc using a monovalent antibody phage display. The absorbance values which evaluate the binding activity of the antibodies to human IgA-Fc in the absence of ATP and adenosine are shown on the vertical axis, and the absorbance values which evaluate the binding activity of the antibodies to 5 human IgA-Fc in the presence of ATP and adenosine are shown on the horizontal axis. Fig. 42 is a figure showing the result of ELISA performed on clone IADL3C5-4_048 obtained from the rationally designed antibody library against human IgA-Fc in the presence or absence of ATP and adenosine at 1 mM. The vertical axis shows the absorbance value which evaluates binding activity of the antibody to human IgA-Fc. Results obtained when using a 0 clone obtained from the rationally designed antibody library and showing binding activity toward human IgA-Fc regardless of the presence of small molecules are presented as the positive control. Results obtained when using the M13KO7 Helper Phage are presented as the negative control. Fig. 43 is a graph showing the binding level (binding response (RU)) when each clone 25 at 1 uM was made to interact for 120 seconds with IL-6R immobilized on Sensorchip CM5 in the presence or absence of each of the small molecules at 1 mM. Fig. 44A shows the ADCC activity of antibodies that bind to antigens in the presence of small molecules toward cells expressing the antigen. It is a figure showing the ADCC activity of clone 6RAD2C1-4_030, which binds to hIL-6R in the presence of ATP, toward CHO cells 30 expressing hIL-6R in the presence (triangles) or absence (circles) of ATP. The open triangles and circles show the measured values, and the filled triangles and circles show the mean values. Fig. 44B shows the ADCC activity of antibodies that bind to antigens in the presence of small molecules toward cells expressing the antigen. It is a figure showing the ADCC activity of clone 6RAD2C1-4_011, which binds to hIL-6R in the presence of ATP, toward CHO cells 35 expressing hIL-6R in the presence (triangles) or absence (circles) of ATP. The open triangles and circles show the measured values, and the filled triangles and circles show the mean values. 2023229507  12 Sep 2023 Fig. 44C shows the ADCC activity of antibodies that bind to antigens in the presence of small molecules toward cells expressing the antigen. It is a figure showing the ADCC activity of MRA, which binds to hIL-6R regardless of the presence or absence of ATP, toward CHO cells expressing in the presence (triangles) or absence (circles) of ATP. The open triangles and 5 circles show the measured values, and the filled triangles and circles show the mean values. Fig. 45 is a figure showing the result of ELISA performed on clone HSADSA1-6_020 obtained from the rationally designed antibody library against HSA in the presence or absence of ATP and adenosine at 10 mM. The vertical axis shows the absorbance value which evaluates binding activity of the antibody to HSA. Results obtained when using a clone obtained from 0 the rationally designed antibody library and showing binding activity toward HSA regardless of the presence of small molecules are presented as the positive control. Results obtained when using the M13KO7 Helper Phage are presented as the negative control. [Mode for Carrying Out the Invention] 5 The definitions and detailed description below are provided to facilitate understanding of the present invention illustrated herein. Amino acids Herein, amino acids are described by one- or three-letter codes or both, for example, 0 Ala / A, Leu / L, Arg / R, Lys / K, Asn / N, Met / M, Asp / D, Phe / F, Cys / C, Pro / P, Gln / Q, Ser / S, Glu / E, Thr / T, Gly / G, Trp / W, His / H, Tyr / Y, Ile / I, or Val / V. Alteration of amino acids For amino acid alteration in the amino acid sequence of an antigen-binding molecule, 25 known methods such as site-directed mutagenesis methods (Kunkel et al. (Proc. Natl. Acad. Sci. USA (1985) 82, 488-492)) and overlap extension PCR may be appropriately employed. Furthermore, several known methods may also be employed as amino acid alteration methods for substitution to non-natural amino acids (Annu. Rev. Biophys. Biomol. Struct. (2006) 35, 225-249; and Proc. Natl. Acad. Sci. U.S.A. (2003) 100 (11), 6353-6357). For example, it is 30 suitable to use a cell-free translation system (Clover Direct (Protein Express)) containing a tRNA which has a non-natural amino acid bound to a complementary amber suppressor tRNA of one of the stop codons, the UAG codon (amber codon). In the present specification, the meaning of the term “and / or” when describing the site of amino acid alteration includes every combination where “and” and “or” are suitably combined. 35 Specifically, for example, “the amino acids at positions 33, 55, and / or 96 are substituted” includes the following variation of amino acid alterations: 2023229507  12 Sep 2023 amino acid(s) at (a) position 33, (b) position 55, (c) position 96, (d) positions 33 and 55, (e) positions 33 and 96, (f) positions 55 and 96, and (g) positions 33, 55, and 96. Furthermore, herein, as an expression showing alteration of amino acids, an expression that shows before and after a number indicating a specific position, one-letter or three-letter 5 codes for amino acids before and after alteration, respectively, may be used appropriately. For example, the alteration N100bL or Asn100bLeu used when substituting an amino acid contained in an antibody variable region indicates substitution of Asn at position 100b (according to Kabat numbering) with Leu. That is, the number shows the amino acid position according to Kabat numbering, the one-letter or three-letter amino-acid code written before the number shows the 0 amino acid before substitution, and the one-letter or three-letter amino-acid code written after the number shows the amino acid after substitution. Similarly the alteration P238D or Pro238Asp used when substituting an amino acid of the Fc region contained in an antibody constant region indicates substitution of Pro at position 238 (according to EU numbering) with Asp. That is, the number shows the amino acid position according to EU numbering, the one-letter or 5 three-letter amino-acid code written before the number shows the amino acid before substitution, and the one-letter or three-letter amino-acid code written after the number shows the amino acid after substitution. Antigens 0 Herein, “antigens” are not particularly limited in their structure, as long as they comprise epitopes to which antigen-binding domains bind. In other words, antigens can be inorganic or organic substances. Other antigens include, for example, the molecules below: 17-IA, 4-1BB, 4Dc, 6-keto-PGF1a, 8-iso-PGF2a, 8-oxo-dG, A1 adenosine receptor, A33, ACE, ACE-2, activin, activin A, activin AB, activin B, activin C, activin RIA, activin RIA ALK-2, 25 activin RIB ALK-4, activin RIIA, activin RIIB, ADAM, ADAM10, ADAM12, ADAM15, ADAM17 / TACE, ADAM8, ADAM9, ADAMTS, ADAMTS4, ADAMTS5, addressin, aFGF, ALCAM, ALK, ALK-1, ALK-7,alpha-1-antitrypsin, alpha-V / beta-1 antagonist, ANG, Ang, APAF-1, APE, APJ, APP, APRIL, AR, ARC, ART, artemin, anti-Id, ASPARTIC, atrial natriuretic peptide, av / b3 integrin, Axl, b2M, B7-1, B7-2, B7-H, B-lymphocyte stimulating factor (BlyS), 30 BACE, BACE-1, Bad, BAFF, BAFF-R, Bag-1, BAK, Bax, BCA-1, BCAM, Bcl, BCMA, BDNF, b-ECGF, bFGF, BID, Bik, BIM, BLC, BL-CAM, BLK, BMP, BMP-2 BMP-2a, BMP-3 Osteogenin, BMP-4 BMP-2b, BMP-5, BMP-6 Vgr-1, BMP-7 (OP-1), BMP-8 (BMP-8a, OP-2), BMPR, BMPR-IA (ALK-3), BMPR-IB (ALK-6), BRK-2, RPK-1, BMPR-II (BRK-3), BMP, b-NGF, BOK, bombesin, bone-derived neurotrophic factor, BPDE, BPDE-DNA, BTC, 35 complement factor 3 (C3), C3a, C4, C5, C5a, C10, CA125, CAD-8, calcitonin, cAMP, carcinoembryonic antigen (CEA), cancer associated antigen, cathepsin A, cathepsin B, cathepsin 2023229507  12 Sep 2023 C / DPPI, cathepsin D, cathepsin E, cathepsin H, cathepsin L, cathepsin O, cathepsin S, cathepsin V, cathepsin X / Z / P, CBL, CCI, CCK2, CCL, CCL1, CCL11, CCL12, CCL13, CCL14, CCL15, CCL16, CCL17, CCL18, CCL19, CCL2, CCL20, CCL21, CCL22, CCL23, CCL24, CCL25, CCL26, CCL27, CCL28, CCL3, CCL4, CCL5, CCL6, CCL7, CCL8, CCL9 / 10, CCR, CCR1, 5 CCR10, CCR10, CCR2, CCR3, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CD1, CD2, CD3, CD3E, CD4, CD5, CD6, CD7, CD8, CD10, CD11a, CD11b, CD11c, CD13, CD14, CD15, CD16, CD18, CD19, CD20, CD21, CD22, CD23, CD25, CD27L, CD28, CD29, CD30, CD30L, CD32, CD33 (p67 protein), CD34, CD38, CD40, CD40L, CD44, CD45, CD46, CD49a, CD52, CD54, CD55, CD56, CD61, CD64, CD66e, CD74, CD80 (B7-1), CD89, CD95, CD123, CD137, 0 CD138, CD140a, CD146, CD147, CD148, CD152, CD164, CEACAM5, CFTR, cGMP, CINC, Botulinum toxin, Clostridium perfringens toxin, CKb8-1, CLC, CMV, CMV UL, CNTF, CNTN-1, COX, C-Ret, CRG-2, CT-1, CTACK, CTGF, CTLA-4, PD1, PDL1, LAG3, TIM3, galectin-9, CX3CL1, CX3CR1, CXCL, CXCL1, CXCL2, CXCL3, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8, CXCL9, CXCL10, CXCL11, CXCL12, CXCL13, CXCL14, CXCL15, 5 CXCL16, CXCR, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, CXCR6,cytokeratin tumor associated antigen, DAN, DCC, DcR3, DC-SIGN, complement regulatory factor (Decay accelerating factor), des (1-3)-IGF-I (brain IGF-1), Dhh, digoxin, DNAM-1, Dnase, Dpp, DPPIV / CD26, Dtk, ECAD, EDA, EDA-A1, EDA-A2, EDAR, EGF, EGFR (ErbB-1), EMA, EMMPRIN, ENA, endothelin receptor, enkephalinase, eNOS, Eot, eotaxin 1, EpCAM, ephrin 0 B2 / EphB4, EPO, ERCC, E-selectin, ET-1, factor IIa, factor VII, factor VIIIc, factor IX, fibroblast activation protein (FAP), Fas, FcR1, FEN-1, ferritin, FGF, FGF-19, FGF-2, FGF3, FGF-8, FGFR, FGFR-3, fibrin, FL, FLIP, Flt-3, Flt-4, follicle stimulating hormone, fractalkine, FZD1, FZD2, FZD3, FZD4, FZD5, FZD6, FZD7, FZD8, FZD9, FZD10, G250, Gas6, GCP-2, GCSF, GD2, GD3, GDF, GDF-1, GDF-3 (Vgr-2), GDF-5 (BMP-14, CDMP-1), GDF-6 (BMP-13, 25 CDMP-2), GDF-7 (BMP-12, CDMP-3), GDF-8 (myostatin), GDF-9, GDF-15 (MIC-1), GDNF, GDNF, GFAP, GFRa-1, GFR-alpha1, GFR-alpha2, GFR-alpha3, GITR, glucagon, Glut4, glycoprotein IIb / IIIa (GPIIb / IIIa), GM-CSF, gp130, gp72, GRO, growth hormone releasing hormone, hapten (NP-cap or NIP-cap), HB-EGF, HCC, HCMV gB envelope glycoprotein, HCMV gH envelope glycoprotein, HCMV UL, hematopoietic growth factor (HGF), Hep B 30 gp120, heparanase, Her2, Her2 / neu (ErbB-2), Her3 (ErbB-3), Her4 (ErbB-4), herpes simplex virus (HSV) gB glycoprotein, HSV gD glycoprotein, HGFA, high molecular weight melanoma-associated antigen (HMW-MAA), HIV gp120, HIV IIIB gp 120 V3 loop, HLA, HLA-DR, HM1.24, HMFG PEM, HRG, Hrk, human cardiac myosin, human cytomegalovirus (HCMV), human growth hormone (HGH), HVEM, I-309, IAP, ICAM, ICAM-1, ICAM-3, ICE, 35 ICOS, IFNg, Ig, IgA receptor, IgE, IGF, IGF binding protein, IGF-1R, IGFBP, IGF-I, IGF-II, IL, IL-1, IL-1R, IL-2, IL-2R, IL-4, IL-4R, IL-5, IL-5R, IL-6, IL-6R, IL-8, IL-9, IL-10, IL-12, IL-13, 2023229507  12 Sep 2023 IL-15, IL-18, IL-18R, IL-21, IL-23, IL-27, interferon (INF)-alpha, INF-beta, INF-gamma, inhibin, iNOS, insulin A chain, insulin B chain, insulin-like growth factor1, integrin alpha2, integrin alpha3, integrin alpha4, integrin alpha4 / beta1, integrin alpha4 / beta7, integrin alpha5 (alpha V), integrin alpha5 / beta1, integrin alpha5 / beta3, integrin alpha6, integrin beta1, integrin 5 beta2,interferon gamma, IP-10, I-TAC, JE, kallikrein 2, kallikrein 5, kallikrein 6, kallikrein 11, kallikrein 12, kallikrein 14, kallikrein 15, kallikrein L1, kallikrein L2, kallikrein L3, kallikrein L4, KC, KDR, keratinocyte growth factor (KGF), laminin 5, LAMP, LAP, LAP (TGF-1), latent TGF-1, latent TGF-1 bp1, LBP, LDGF, LECT2, lefty, Lewis-Y antigen, Lewis-Y associated antigen, LFA-1, LFA-3, Lfo, LIF, LIGHT, lipoprotein, LIX, LKN, Lptn, L-selectin, LT-a, LT-b, 0 LTB4, LTBP-1, lung surface, luteinizing hormone, lymphotoxin beta receptor, Mac-1, MAdCAM, MAG, MAP2, MARC, MCAM, MCAM, MCK-2, MCP, M-CSF, MDC, Mer, METALLOPROTEASES, MGDF receptor, MGMT, MHC (HLA-DR), MIF, MIG, MIP, MIP-1-alpha, MK, MMAC1, MMP, MMP-1, MMP-10, MMP-11, MMP-12, MMP-13, MMP-14, MMP-15, MMP-2, MMP-24, MMP-3, MMP-7, MMP-8, MMP-9, MPIF, Mpo, MSK, MSP, 5 mucin (Muc1), MUC18, Mullerian-inhibiting substance, Mug, MuSK, NAIP, NAP, NCAD, N-C adherin, NCA 90, NCAM, NCAM, neprilysin, neurotrophin-3, -4, or -6, neurturin, nerve growth factor (NGF), NGFR, NGF-beta, nNOS, NO, NOS, Npn, NRG-3, NT, NTN, OB, OGG1, OPG, OPN, OSM, OX40L, OX40R, p150, p95, PADPr, parathyroid hormone, PARC, PARP, PBR, PBSF, PCAD, P-cadherin, PCNA, PDGF, PDGF, PDK-1, PECAM, PEM, PF4, PGE, PGF, PGI2, 0 PGJ2, PIN, PLA2, placental alkaline phosphatase (PLAP), PlGF, PLP, PP14, proinsulin, prorelaxin, protein C, PS, PSA, PSCA, prostate-specific membrane antigen (PSMA), PTEN, PTHrp, Ptk, PTN, R51, RANK, RANKL, RANTES, RANTES, relaxin A chain, relaxin B chain, renin, respiratory syncytial virus (RSV) F, RSV Fgp, Ret, Rheumatoid factor, RLIP76, RPA2, RSK, S100, SCF / KL, SDF-1, SERINE, serum albumin, sFRP-3, Shh, SIGIRR, SK-1, SLAM, 25 SLPI, SMAC, SMDF, SMOH, SOD, SPARC, Stat, STEAP, STEAP-II, TACE, TACI, TAG-72 (tumor-associated glycoprotein-72), TARC, TCA-3, T-cell receptor (for example, T-cell receptor alpha / beta), TdT, TECK, TEM1, TEM5, TEM7, TEM8, TERT, testis PLAP-like alkaline phosphatase, TfR, TGF, TGF-alpha, TGF-beta, TGF-beta Pan Specific, TGF-betaRI (ALK-5), TGF-betaRII, TGF-betaRIIb, TGF-betaRIII, TGF-beta1, TGF-beta2, TGF-beta3, TGF-beta4, 30 TGF-beta5, thrombin, thymus Ck-1, thyroid-stimulating hormone, Tie, TIMP, TIQ, tissue factor, TMEFF2, Tmpo, TMPRSS2, TNF, TNF-alpha, TNF-alphabeta, TNF-beta2, TNFc, TNF-RI, TNF-RII, TNFRSF10A (TRAIL R1 Apo-2, DR4), TNFRSF10B (TRAIL R2 DR5, KILLER, TRICK-2A, TRICK-B), TNFRSF10C (TRAIL R3 DcR1, LIT, TRID), TNFRSF10D (TRAIL R4 DcR2, TRUNDD), TNFRSF11A (RANK ODF R, TRANCE R), TNFRSF11B (OPG OCIF, TR1), 35 TNFRSF12 (TWEAK R FN14), TNFRSF13B (TACI), TNFRSF13C (BAFF R), TNFRSF14 (HVEM ATAR, HveA, LIGHT R, TR2), TNFRSF16 (NGFR p75NTR), TNFRSF17 (BCMA), 2023229507  12 Sep 2023 TNFRSF18 (GITR AITR), TNFRSF19 (TROY TAJ, TRADE), TNFRSF19L (RELT), TNFRSF1A (TNF RI CD120a, p55-60), TNFRSF1B (TNF RII CD120b, p75-80), TNFRSF26 (TNFRH3) , TNFRSF3 (LTbR TNF RIII, TNFC R), TNFRSF4 (OX40 ACT35, TXGP1 R), TNFRSF5 (CD40 p50), TNFRSF6 (Fas Apo-1, APT1, CD95), TNFRSF6B (DcR3 M68, TR6), 5 TNFRSF7 (CD27), TNFRSF8 (CD30), TNFRSF9 (4-1BB CD137, ILA), TNFRSF21 (DR6), TNFRSF22 (DcTRAIL R2 TNFRH2), TNFRST23 (DcTRAIL R1 TNFRH1), TNFRSF25 (DR3 Apo-3, LARD, TR-3, TRAMP, WSL-1), TNFSF10 (TRAIL Apo-2 ligand, TL2), TNFSF11 (TRANCE / RANK ligand ODF, OPG ligand), TNFSF12 (TWEAK Apo-3 ligand, DR3 ligand), TNFSF13 (APRIL TALL2), TNFSF13B (BAFF BLYS, TALL1, THANK, TNFSF20), TNFSF14 0   (LIGHT HVEM ligand, LTg), TNFSF15 (TL1A / VEGI), TNFSF18 (GITR ligand AITR ligand, TL6), TNFSF1A (TNF-a Conectin, DIF, TNFSF2), TNFSF1B (TNF-b LTa, TNFSF1), TNFSF3 (LTb TNFC, p33), TNFSF4 (OX40 ligand gp34, TXGP1), TNFSF5 (CD40 ligand CD154, gp39, HIGM1, IMD3, TRAP), TNFSF6 (Fas ligand Apo-1 ligand, APT1 ligand), TNFSF7 (CD27 ligand CD70), TNFSF8 (CD30 ligand CD153), TNFSF9 (4-1BB ligand CD137 ligand), TP-1, 5 t-PA, Tpo, TRAIL, TRAIL R, TRAIL-R1, TRAIL-R2, TRANCE, transferrin receptor, TRF, Trk, TROP-2, TLR1 (Toll-like receptor 1), TLR2, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8, TLR9, TLR10, TSG, TSLP, tumor associated antigen CA125, tumor associated antigen expressing Lewis-Y associated carbohydrates, TWEAK, TXB2, Ung, uPAR, uPAR-1, urokinase, VCAM, VCAM-1, VECAD, VE-Cadherin, VE-cadherin-2, VEFGR-1 (flt-1), VEGF, VEGFR, VEGFR-3 0 (flt-4), VEGI, VIM, virus antigen, VLA, VLA-1, VLA-4, VNR integrin, von Willebrand factor, WIF-1, WNT1, WNT2, WNT2B / 13, WNT3, WNT3A, WNT4, WNT5A, WNT5B, WNT6, WNT7A, WNT7B, WNT8A, WNT8B, WNT9A, WNT9A, WNT9B, WNT10A, WNT10B, WNT11, WNT16, XCL1, XCL2, XCR1, XCR1, XEDAR, XIAP, XPD, HMGB1, IgA, Ap, CD81, CD97, CD98, DDR1, DKK1, EREG, Hsp90, IL-17 / IL-17R, IL-20 / IL-20R, oxidized LDL, 25 PCSK9, prekallikrein, RON, TMEM16F, SOD1, Chromogranin A, Chromogranin B, tau, VAP1, high molecular weight kininogen, IL-31, IL-31R, Nav1.1, Nav1.2, Nav1.3, Nav1.4, Nav1.5, Nav1.6, Nav1.7, Nav1.8, Nav1.9, EPCR, C1, C1q, C1r, C1s, C2, C2a, C2b, C3, C3a, C3b, C4, C4a, C4b, C5, C5a, C5b, C6, C7, C8, C9, factor B, factor D, factor H, properdin, sclerostin, fibrinogen, fibrin, prothrombin, thrombin, tissue factor, factor V, factor Va, factor VII, factor 30 VIIa, factor VIII, factor VIIIa, factor IX, factor IXa, factor X, factor Xa, factor XI, factor XIa, factor XII, factor XIIa, factor XIII, factor XIIIa, TFPI, antithrombin III, EPCR, thrombomodulin, TAPI, tPA, plasminogen, plasmin, PAI-1, PAI-2, GPC3, Syndecan-1, Syndecan-2, Syndecan-3, Syndecan-4, LPA, and S1P; and receptors for hormone and growth factors. Preferred antigens are antigens that are expressed in cancer cells, immune cells, stromal cells, or such present in 35 cancer tissues or inflammatory tissues. While receptors are recited as examples of the above-mentioned antigens, when these 2023229507  12 Sep 2023 receptors exist in soluble forms in biological fluids, they may be used as antigens that bind to the antigen-binding molecule of the present invention, which contains an antigen-binding domain whose antigen-binding activity varies depending on the concentration of the target tissue-specific compound. An example of a non-limiting embodiment of such a soluble receptor is the soluble 5 IL-6R, which is a protein consisting of the amino acids at positions 1 to 357 in the IL-6R polypeptide sequence of SEQ ID NO: 1 as described in Mullberg et al. (J. Immunol. (1994) 152 (10), 4958-4968). Membrane-type molecules expressed on cell membranes and soluble molecules secreted from cells to the outside of the cells are included in the examples of the above-mentioned 0 antigens. When the antigen-binding molecule of the present invention, which contains an antigen-binding domain whose antigen-binding activity varies depending on the concentration of the target tissue-specific compound, binds to a soluble molecule secreted from cells, it is preferable that the antigen-binding molecule has neutralizing activity as described later. The fluids in which the soluble molecules exist are not limited, and the soluble 5 molecules may exist in biological fluids, or more specifically in all fluids filling the space between tissues and cells or vessels in organisms. In a non-limiting embodiment, the soluble molecules to which antigen-binding molecules of the present invention bind may be present in the extracellular fluid. In vertebrates, extracellular fluid is a general term for plasma, interstitial fluid, lymph, compact connective tissue, cerebrospinal fluid, spinal fluid, puncture fluid, 0 synovial fluid, or such components in the bone and cartilage, alveolar fluid (bronchoalveolar lavage fluid), peritoneal fluid, pleural fluid, pericardial effusion, cyst fluid, aqueous humor (hydatoid), or such transcellular fluids (various fluids in the glandular cavities and fluids in the digestive tract cavity and other body cavity fluids produced as a result of active transport / secretory activities of cells). 25 When an antigen-binding molecule of the present invention comprising an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a target tissue-specific compound binds to a membrane-type molecule expressed on a cell membrane, suitable examples of the antigen-binding molecule include antigen-binding molecules which have cytotoxic activity, bind to a cytotoxic substance, or have the ability to bind 30 to a cytotoxic substance, as described later. Furthermore, antigen-binding molecules having a neutralizing activity instead of the properties of having a cytotoxic activity, binding to a cytotoxic substance, or having the ability to bind to a cytotoxic substance; or in addition to these properties are also suitable examples of a non-limiting embodiment. 35 Epitopes “Epitope” means an antigenic determinant in an antigen, and refers to an antigen site to 2023229507  12 Sep 2023 which the antigen-binding domain of an antigen-binding molecule disclosed herein binds. Thus, for example, the epitope can be defined according to its structure. Alternatively, the epitope may be defined according to the antigen-binding activity of an antigen-binding molecule that recognizes the epitope. When the antigen is a peptide or polypeptide, the epitope can be 5 specified by the amino acid residues forming the epitope. Alternatively, when the epitope is a sugar chain, the epitope can be specified by its specific sugar chain structure. A linear epitope is an epitope that contains an epitope whose primary amino acid sequence has been recognized. Such a linear epitope typically contains at least three and most commonly at least five, for example, about 8 to about 10 or 6 to 20 amino acids in a specific 0 sequence. In contrast to the linear epitope, a “conformational epitope” is an epitope in which the primary amino acid sequence containing the epitope is not the only determinant of the recognized epitope (for example, the primary amino acid sequence of a conformational epitope is not necessarily recognized by an epitope-defining antibody). Conformational epitopes may 5 contain a greater number of amino acids compared to linear epitopes. A conformational epitope-recognizing antibody recognizes the three-dimensional structure of a peptide or protein. For example, when a protein molecule folds and forms a three-dimensional structure, amino acids and / or polypeptide main chains that form a conformational epitope become aligned, and the epitope is made recognizable by the antibody. Methods for determining epitope 0 conformations include, for example, X ray crystallography, two-dimensional nuclear magnetic resonance, site-specific spin labeling, and electron paramagnetic resonance, but are not limited thereto. See, for example, Epitope Mapping Protocols in Methods in Molecular Biology (1996), Vol. 66, Morris (ed.). The structure of the antigen-binding domain which binds to an epitope is called a 25 paratope. An epitope and a paratope bind with stability through the action of hydrogen bonds, electrostatic force, van der Waals force, hydrophobic bonds, and such between the epitope and the paratope. This strength of binding between the epitope and paratope is called affinity. The total sum of binding strength when a plurality of antigens and a plurality of antigen-binding molecules bind is referred to as avidity. When an antibody comprising a plurality of 30 antigen-binding domains (i.e., multivalent antibody) or such binds to a plurality of epitopes, the affinity acts synergistically, and therefore avidity becomes higher than affinity. Binding Activity Examples of a method for assessing the epitope binding by a test antigen-binding 35 molecule containing an IL-6R antigen-binding domain are described below. According to the examples below, methods for assessing the epitope binding by a test antigen-binding molecule 2023229507  12 Sep 2023 containing an antigen-binding domain for an antigen other than IL-6R, can also be appropriately conducted. For example, whether a test antigen-binding molecule containing an IL-6R antigen-binding domain recognizes a linear epitope in the IL-6R molecule can be confirmed for 5 example as mentioned below. A linear peptide comprising an amino acid sequence forming the extracellular domain of IL-6R is synthesized for the above purpose. The peptide can be synthesized chemically, or obtained by genetic engineering techniques using a region encoding the amino acid sequence corresponding to the extracellular domain in an IL-6R cDNA. Then, a test antigen-binding molecule containing an IL-6R antigen-binding domain is assessed for its 0 binding activity towards a linear peptide comprising the amino acid sequence forming the extracellular domain. For example, an immobilized linear peptide can be used as an antigen by ELISA to evaluate the binding activity of the antigen-binding molecule towards the peptide. Alternatively, the binding activity towards a linear peptide can be assessed based on the level that the linear peptide inhibits the binding of the antigen-binding molecule to IL-6R-expressing 5 cells. These tests can demonstrate the binding activity of the antigen-binding molecule towards the linear peptide. Whether a test antigen-binding molecule containing an IL-6R antigen-binding domain recognizes a conformational epitope can be assessed as follows. IL-6R-expressing cells are prepared for the above purpose. A test antigen-binding molecule containing an IL-6R 0 antigen-binding domain can be determined to recognize a conformational epitope when it strongly binds to IL-6R-expressing cells upon contact, but does not substantially bind to an immobilized linear peptide comprising an amino acid sequence forming the extracellular domain of IL-6R. Herein, “not substantially bind” means that the binding activity is 80% or less, generally 50% or less, preferably 30% or less, and particularly preferably 15% or less compared 25 to the binding activity towards cells expressing human IL-6R. Methods for assaying the binding activity of a test antigen-binding molecule containing an IL-6R antigen-binding domain towards IL-6R-expressing cells include, for example, the methods described in Antibodies: A Laboratory Manual (Ed Harlow, David Lane, Cold Spring Harbor Laboratory (1988) 359-420). Specifically, the assessment can be performed based on 30 the principle of ELISA or fluorescence activated cell sorting (FACS) using IL-6R-expressing cells as antigen. In the ELISA format, the binding activity of a test antigen-binding molecule containing an IL-6R antigen-binding domain towards IL-6R-expressing cells can be assessed quantitatively by comparing the levels of signal generated by enzymatic reaction. Specifically, a test 35 polypeptide complex is added to an ELISA plate onto which IL-6R-expressing cells are immobilized. Then, the test antigen-binding molecule bound to the cells is detected using an 2023229507  12 Sep 2023 enzyme-labeled antibody that recognizes the test antigen-binding molecule. Alternatively, when FACS is used, a dilution series of a test antigen-binding molecule is prepared, and the antibody binding titer for IL-6R-expressing cells can be determined to compare the binding activity of the test antigen-binding molecule towards IL-6R-expressing cells. 5 The binding of a test antigen-binding molecule towards an antigen expressed on the surface of cells suspended in buffer or the like can be detected using a flow cytometer. Known flow cytometers include, for example, the following devices: FACSCantoTM II FACSAriaTM 0  FACSArrayTM FACSVantageTM SE FACSCaliburTM (all are trade names of BD Biosciences) EPICS ALTRA HyPerSort Cytomics FC 500 5 EPICS XL-MCL ADC EPICS XL ADC Cell Lab Quanta / Cell Lab Quanta SC (all are trade names of Beckman Coulter). Preferable methods for assaying the binding activity of a test antigen-binding molecule containing an IL-6R antigen-binding domain towards an antigen include, for example, the following method. First, IL-6R-expressing cells are reacted with a test antigen-binding 0 molecule, and then this is stained with an FITC-labeled secondary antibody that recognizes the antigen-binding molecule. The test antigen-binding molecule is appropriately diluted with a suitable buffer to prepare the molecule at a desired concentration. For example, the molecule can be used at a concentration within the range of 10 Ligml to 10 ng / ml. Then, the fluorescence intensity and cell count are determined using FACSCalibur (BD). The fluorescence intensity 25 obtained by analysis using the CELL QUEST Software (BD), i.e., the Geometric Mean value, reflects the quantity of antibody bound to cells. That is, the binding activity of a test antigen-binding molecule, which is represented by the quantity of the test antigen-binding molecule bound, can be determined by measuring the Geometric Mean value. Whether a test antigen-binding molecule containing an IL-6R antigen-binding domain 30 shares a common epitope with another antigen-binding molecule can be assessed based on the competition between the two molecules for the same epitope. The competition between antigen-binding molecules can be detected by cross-blocking assay or the like. For example, the competitive ELISA assay is a preferred cross-blocking assay. Specifically, in cross-blocking assay, the IL-6R protein immobilized to the wells of a 35 microtiter plate is pre-incubated in the presence or absence of a candidate competitor antigen-binding molecule, and then a test antigen-binding molecule is added thereto. The 2023229507  12 Sep 2023 quantity of test antigen-binding molecule bound to the IL-6R protein in the wells is indirectly correlated with the binding ability of a candidate competitor antigen-binding molecule that competes for the binding to the same epitope. That is, the greater the affinity of the competitor antigen-binding molecule for the same epitope, the lower the binding activity of the test 5 antigen-binding molecule towards the IL-6R protein-coated wells. The quantity of the test antigen-binding molecule bound to the wells via the IL-6R protein can be readily determined by labeling the antigen-binding molecule in advance. For example, a biotin-labeled antigen-binding molecule is measured using an avidin / peroxidase conjugate and appropriate substrate. In particular, cross-blocking assay that uses enzyme labels 0 such as peroxidase is called “competitive ELISA assay”. The antigen-binding molecule can also be labeled with other labeling substances that enable detection or measurement. Specifically, radiolabels, fluorescent labels, and such are known. When the candidate competitor antigen-binding molecule can block the binding by a test antigen-binding molecule containing an IL-6R antigen-binding domain by at least 20%, 5 preferably at least 20 to 50%, and more preferably at least 50% compared to the binding activity in a control experiment conducted in the absence of the competitor antigen-binding molecule, the test antigen-binding molecule is determined to substantially bind to the same epitope bound by the competitor antigen-binding molecule, or compete for the binding to the same epitope. When the structure of an epitope bound by a test antigen-binding molecule containing 0 an IL-6R antigen-binding domain has already been identified, whether the test and control antigen-binding molecules share a common epitope can be assessed by comparing the binding activities of the two antigen-binding molecules towards a peptide prepared by introducing amino acid mutations into the peptide forming the epitope. To measure the above binding activities, for example, the binding activities of test and 25 control antigen-binding molecules towards a linear peptide into which a mutation is introduced are compared in the above ELISA format. Besides the ELISA methods, the binding activity towards the mutant peptide bound to a column can be determined by flowing test and control antigen-binding molecules in the column, and then quantifying the antigen-binding molecule eluted in the elution solution. Methods for adsorbing a mutant peptide to a column, for 30 example, in the form of a GST fusion peptide, are known. Alternatively, when the identified epitope is a conformational epitope, whether test and control antigen-binding molecules share a common epitope can be assessed by the following method. First, IL-6R-expressing cells and cells expressing IL-6R with a mutation introduced into the epitope are prepared. The test and control antigen-binding molecules are added to a 35   cell suspension prepared by suspending these cells in an appropriate buffer such as PBS. Then, the cell suspensions are appropriately washed with a buffer, and an FITC-labeled antibody that 2023229507  12 Sep 2023 recognizes the test and control antigen-binding molecules is added thereto. The fluorescence intensity and number of cells stained with the labeled antibody are determined using FACSCalibur (BD). The test and control antigen-binding molecules are appropriately diluted using a suitable buffer, and used at desired concentrations. For example, they may be used at a 5 concentration within the range of 10 ugml to 10 ng / ml. The fluorescence intensity determined by analysis using the CELL QUEST Software (BD), i.e., the Geometric Mean value, reflects the quantity of labeled antibody bound to cells. That is, the binding activities of the test and control antigen-binding molecules, which are represented by the quantity of labeled antibody bound, can be determined by measuring the Geometric Mean value. 0 In the above method, whether an antigen-binding molecule does “not substantially bind to cells expressing mutant IL-6R” can be assessed, for example, by the following method. First, the test and control antigen-binding molecules bound to cells expressing mutant IL-6R are stained with a labeled antibody. Then, the fluorescence intensity of the cells is determined. When FACSCalibur is used for fluorescence detection by flow cytometry, the determined 5 fluorescence intensity can be analyzed using the CELL QUEST Software. From the Geometric Mean values in the presence and absence of the polypeptide complex, the comparison value (AGeo-Mean) can be calculated according to Formula 1 below to determine the ratio of increase in fluorescence intensity as a result of the binding by the antigen-binding molecule. 0 Formula 1: AGeo-Mean = Geo-Mean (in the presence of the polypeptide complex) / Geo-Mean (in the absence of the polypeptide complex) The Geometric Mean comparison value (AGeo-Mean value for the mutant IL-6R 25 molecule) determined by the above analysis, which reflects the quantity of a test antigen-binding molecule bound to cells expressing mutant IL-6R, is compared to the AGeo-Mean comparison value that reflects the quantity of the test antigen-binding molecule bound to IL-6R-expressing cells. In this case, the concentrations of the test antigen-binding molecule used to determine the AGeo-Mean comparison values for IL-6R-expressing cells and cells expressing mutant IL-6R are 30 particularly preferably adjusted to be equal or substantially equal. An antigen-binding molecule that has been confirmed to recognize an epitope in IL-6R is used as a control antigen-binding molecule. If the AGeo-Mean comparison value of a test antigen-binding molecule for cells expressing mutant IL-6R is smaller than the AGeo-Mean comparison value of the test 35 antigen-binding molecule for IL-6R-expressing cells by at least 80%, preferably 50%, more preferably 30%, and particularly preferably 15%, then the test antigen-binding molecule “does 2023229507  12 Sep 2023 not substantially bind to cells expressing mutant IL-6R”. The formula for determining the Geo-Mean (Geometric Mean) value is described in the CELL QUEST Software User’s Guide (BD biosciences). When the comparison shows that the comparison values are substantially equivalent, the epitope for the test and control antigen-binding molecules can be determined to 5 be the same. Target Tissue The term "target tissue" as used herein refers to a tissue containing cells carrying antigens to which the antigen-binding molecules of the present invention bind in a manner 0 dependent on compounds. It is a tissue that yields positive pharmacological effects for the organism carrying the tissue, when the antigen-binding molecules bind to a membrane-type molecule expressed on the cells or bind to a soluble molecule present in the tissue. In this case, the phrase "positive pharmacological effects" refers to effects that relieve, alleviate, ameliorate, or cure symptoms brought about by pathological sites containing the target tissue for the 5 organism carrying the tissue. When the symptoms are brought about by malignant tumors such as cancer, a non-limiting embodiment of a mechanism that yields such a pharmacological effect is, for example, cytotoxic activity and growth inhibition against cancer cells, and immunostimulation in cancer tissues. In the case of inflammatory diseases, examples of such a non-limiting embodiment of the mechanism include immunosuppression and activity to block 0 actions of inflammatory cytokines in inflammatory tissues. Cancer tissue-specific compounds The term "compound specific to a cancer tissue (cancer tissue-specific compound)" as used herein refers to a compound differentially present in cancer tissues as compared to 25 non-cancerous tissues. Herein, the term "cancer" is generally used to describe malignant neoplasms, which may be metastatic or non-metastatic. Non-limiting examples of carcinomas developed from epithelial tissues such as skin or digestive tract include brain tumor, skin cancer, head and neck cancer, esophageal cancer, lung cancer, stomach cancer, duodenal cancer, breast cancer, prostate cancer, cervical cancer, endometrial cancer, pancreatic cancer, liver cancer, 30 colorectal cancer, colon cancer, bladder cancer, and ovarian cancer. Non-limiting examples of sarcomas developed from non-epithelial (interstitial) tissues such as muscles include osteosarcoma, chondrosarcoma, rhabdomyosarcoma, leiomyosarcoma, liposarcoma, and angiosarcoma. Non-limiting examples of hematological cancer derived from hematopoietic organs include malignant lymphomas including Hodgkin's lymphoma and non Hodgkin's 35 lymphoma; leukemia including acute myelocytic leukemia or chronic myelocytic leukemia, and acute lymphatic leukemia or chronic lymphatic leukemia; and multiple myeloma. The term 2023229507  12 Sep 2023 "neoplasm" widely used herein refers to any newly formed diseased tissue tumor. In the present invention, neoplasms cause formation of tumors, which are partly characterized by angiogenesis. Neoplasms may be benign such as hemangioma, glioma, or teratoma, or malignant such as carcinoma, sarcoma, glioma, astrocytoma, neuroblastoma, or retinoblastoma. 5 The term "cancer tissue" refers to a tissue containing at least one cancer cell. Therefore, as cancer tissues contain cancer cells and blood vessels, it refers to all cell types contributing to the formation of a tumor mass containing cancer cells and endothelial cells. Herein, "tumor mass" refers to a foci of tumor tissue. The term "tumor" is generally used to mean a benign neoplasm or a malignant neoplasm. 0 For example, in several embodiments, cancer tissue-specific compounds may be compounds defined by qualitative properties of cancer tissues such as being present in cancer tissues but absent in non-cancer tissues, or being absent in cancer tissues but present in non-cancer tissues. In other embodiments, cancer tissue-specific compounds may be compounds defined by quantitative properties of cancer tissues such as being present in cancer 5 tissues at a concentration different (for example, higher concentration or lower concentration) from that in non-cancer tissues. For example, cancer tissue-specific compounds are present differentially at arbitrary concentrations. Generally, cancer tissue-specific compounds can be present at a concentration increased by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least40%, at least 45%, at least 50%, at least 55%, at 0 least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 2-fold, at least 5-fold, at least 10-fold, at least 50-fold, at least 100-fold, at least 103-fold, at least 104-fold, at least 105-fold, at least 106-fold, or more, or up to infinity (i.e., when the compound is absent in non-cancerous tissues). Alternatively, they can generally be present 25 at a concentration decreased by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, or at least 100% (i.e., absent). Preferably, cancer tissue-specific compounds are differentially present at statistically significant concentrations (that is, as determined using either Welch's t-test or 30 Wilcoxon rank sum test, the p value is less than 0.05 and / or the q value is less than 0.10). Examples of a non-limiting embodiment of a cancer tissue-specific compound include compounds which are cancer tissue-specific metabolites produced by metabolic activities characteristic of cancer cells, immune cells, or stromal cells contained in cancer tissues, such as those described below (cancer tissue-specific metabolites, cancer cell-specific metabolites, 35 metabolites specific to immune cells that infiltrated into cancer tissues, and cancer stromal cell-specific metabolites). 2023229507  12 Sep 2023 Cancer tissue-specific metabolites The term "metabolism" refers to chemical changes that take place in biological tissues and includes "anabolism" and "catabolism". Anabolism refers to biosynthesis or accumulation 5 of molecules, and catabolism refers to degradation of molecules. "Metabolites" are intermediates or products that arise from metabolism. "Primary metabolites" refers to metabolites directly involved in the process of growth or proliferation of cells or organisms. "Secondary metabolites" refer to products that are not directly involved in such process of growth or proliferation, and are products such as pigments or antibiotics that are produced as a 0 result of metabolism which biosynthesizs substances that are not directly involved in biological phenomena common to cells and organisms. The metabolites may be metabolites of "biopolymers", or they may be metabolites of "small molecules". "Biopolymers" are polymers comprising one or more types of repeating units. Biopolymers are generally found in biological systems, and examples include cells forming the organism and intercellular matrices that adhere 5 to them, molecules having a molecular weight of approximately 5000 or more which form structures such as interstitial matrices, particularly polysaccharides (carbohydrates and such), peptides (this term is used so as to include polypeptides and proteins), and polynucleotides, and similarly their analogs such as compounds composed of or including amino acid analogs or non-amino acid groups. "Small molecules" refers to natural chemical substances other than 0 "biopolymers" that exist in vivo. Suitable examples of a non-limiting embodiment of a cancer tissue-specific metabolite described herein include cancer cell-specific small-molecule metabolites (Eva Gottfried, Katrin Peter and Marina P. Kreutz, From Molecular to Modular Tumor Therapy (2010) 3 (2), 111-132). In addition, metabolites that are highly produced by immune cells that infiltrate into cancer tissues, and metabolites that are highly produced by 25 stromal cells that support the survival and / or growth of cancer cells (cancer stromal cells or cancer associated stromal fibroblasts (CAF)) are also included. Infiltrating immune cells are, for example, dendritic cells, inhibitory dendritic cells, inhibitory T cells, exhausted T cells, and myeloma derived suppressor cells (MDSC). Furthermore, metabolites of the present invention include compounds released from inside the cells to outside the cells when cells present in cancer 30 tissues (cancer cells, immune cells, or stromal cells) die due to apoptosis, necrosis, or such. To identify cancer cell-specific metabolites, metabolomic analyses focused on metabolic profiling can be suitably used, in addition to transcriptome-level analyses (for example, Dhanasekaran et al. (Nature (2001) 412, 822-826), Lapointe et al. (Proc. Natl. Acad. Sci. U.S.A. (2004) 101, 811-816) or Perou et al. (Nature (2000) 406, 747-752)) and proteome-level analyses 35 (for example, Ahram et al. (Mol. Carcinog. (2002) 33, 9-15), Hood et al. (Mol. Cell. Proteomics (2005) 4, 1741-1753)). More specifically, to identify metabolites in test samples, metabolic 2023229507  12 Sep 2023 profiling that uses high-pressure liquid chromatography (HPLC), nuclear magnetic resonance (NMR) (Brindle et al. (J. Mol. Recognit. (1997) 10, 182-187), mass spectrometry (Gates and Sweeley (Clin. Chem. (1978) 24, 1663-1673) (GC / MS and LC / MS)), and ELISA or such individually and / or in combination may be used appropriately. 5 These studies elucidated heterogeneity within the constituted tumors which results from changing the concentration gradient of growth factors and metabolites (glucose, oxygen, or such) that enable cancer cell growth under low oxygen pressure conditions (Dang and Semenza (Trends Biochem. Sci. (1999) 24, 68-72)). In these studies, cell line models are also used to understand the change in energy utilization pathway depending on the different malignancy 0 levels of tumors (Vizan et al. (Cancer Res. (2005) 65, 5512-5515)). Examples of a non-limiting embodiment of the technical components of the metabolomics platform include sample extraction, separation, detection, spectroscopic analysis, data normalization, description of class-specific metabolites, pathway mapping, confirmation, and functional characterization of candidate metabolites described by Lawton et al. (Pharmacogenomics (2008) 9, 383). These 5 methods enable identification of cancer cell-specific metabolites in desired cancer tissues. Examples of a non-limiting embodiment of cancer tissue-specific compounds or cancer tissue-specific metabolites used in the present invention preferably include at least one compound selected from the compounds below. At least one compound means that in addition to cases where the antigen-binding activity of a same antigen-binding domain described below 0 depends on one type of cancer tissue-specific compound or metabolite, cases where it depends on several types of cancer tissue-specific compounds or metabolites are included. (1) Primary metabolites of the Krebs cycle or of the glycolytic system such as lactic acid, succinic acid, and citric acid 25 Preferable examples of a non-limiting embodiment of a cancer tissue-specific compound, particularly a cancer cell-specific metabolite, used in the present invention include primary metabolites such as lactic acid, succinic acid, and citric acid, which are produced as a result of glucose metabolism, and are present at higher concentrations in cancer tissues as compared to in the surrounding non-cancerous tissues. The glycolytic system phenotype, 30 which is characterized as an up-regulation of enzymes of the glycolytic system (Embden-Meyerhof pathway) such as pyruvate kinase, hexokinase, and lactic acid dehydrogenase (LDH), has been conventionally known to be a characteristic of solid tumors as Warburg effect. That is, in tumor cells, high expression of the pyruvate kinase isoform M2 which is 35 necessary for anaerobic glycolysis, and not isoform M1, is considered to be working advantageously for the growth of tumor cells in vivo (Christofk et al. (Nature (2008) 452, 2023229507  12 Sep 2023 230-233). Pyruvic acid produced by pyruvate kinase is subjected to feedback inhibition by lactic acid produced as a result of equilibrium reaction by lactic acid dehydrogenase (LDH) under anaerobic conditions. Since the feedback inhibition causes promotion of respiration in mitochondria (Krebs cycle) and cell growth inhibition, up regulation of LDH, hexokinase, and 5 glucose transporter (GLUT) is said to play an important role in the proliferation of cancer cells (Fantin et al. (Cancer Cell (2006) 9, 425-434)). Glucose is metabolized by the glycolytic system, and the final metabolite lactic acid is transported together with protons to the tumor surrounding, and as a result, the pH of the tissues surrounding the tumor is said to become acidic. Lactic acid, which is the final product of the glycolytic pathway, as well as succinic acid and 0 citric acid produced by promotion of respiration in mitochondria are known to be accumulated in cancer tissues (Teresa et al. (Mol. Cancer (2009) 8, 41-59)). Examples of a non-limiting embodiment of cancer tissue-specific compounds, particularly cancer cell-specific metabolites, used in the present invention preferably include such primary metabolites such as lactic acid, succinic acid, and citric acid produced by metabolism by the glycolytic pathway. Furthermore, 5 succinic acid which is present at high concentration in cells is known to leak out to the outside of the cells upon cell death (Nature Immunology, (2008) 9, 1261-1269). Therefore, succinic acid concentration is thought to be increased in cancer tissues in which cell death occurs frequently. (2) Amino acids such as alanine, glutamic acid, and aspartic acid 0 Besides the above-mentioned glucose metabolism, the amino acid metabolism is also known to be altered in tumor cells which require continuous supply of essential amino acids and non-essential amino acids that are necessary for the biosynthesis of biopolymers under anaerobic conditions. Glutamine which contains two nitrogens in its side chain acts as a nitrogen transporter, and is an amino acid that is most widely distributed in an organism. Tumor cells, in 25 which the rate of glutamine uptake into cells is increased, is said to be functioning as a glutamine trap. Such increase in the uptake of glutamine and activity of converting into glutamic acid and lactic acid is called "glutaminolysis", and is considered to be a characteristic of transformed (tumor) cells (Mazurek and Eigenbrodt (Anticancer Res. (2003) 23, 1149-1154); and Mazurek et al. (J. Cell. Physiol. (1999) 181, 136-146)). As a result, cancer patients show an increase in 30 glutamic acid concentration while showing a decrease in plasma glutamine level (Droge et al. (Immunobiology (1987) 174, 473-479)). Furthermore, correlation was observed between concentrations of 13C-labeled succinic acid, 13C-labeled alanine, 13C-labeled glutamic acid, and 13C-labeled citric acid in studies on 13C-radiolabeled glucose metabolism in lung cancer tissues. Suitable examples of a non-limiting embodiment of cancer tissue-specific compounds used in 35 this invention include alanine, glutamic acid, and aspartic acid which accumulate at high concentrations in cancer tissues through such glutaminolysis and the like. 2023229507  12 Sep 2023 (3) Amino acid metabolite such as kynurenine Indolamine 2,3-dioxygenase (IDO) is a tryptophan-metabolizing enzyme which is highly expressed in many cancers such as melanoma, colon cancer, and kidney cancer 5 (Uyttenhove et al. (Nat. Med. (2003) 9, 1269-127)); and it is known to have two isoforms (Lob et al. (Cancer Immunol. Immunother. (2009) 58, 153-157)). IDO catalyzes the conversion of tryptophan to kynurenine (shown as Compound 1), and is the first enzyme in the nicotinamide nucleotide (NAD) de novo pathway. Furthermore, in glioma which does not express IDO, kynurenine is produced from tryptophan by tryptophan 2,3-dioxygenase (TDO) in the liver 0 (Opitz et al. (Nature (2011) 478, 7368, 197-203)). IDO is also expressed in dendritic cells infiltrated into cancer tissues, and dendritic cells also produce kynurenine (J. Immunol. (2008) 181, 5396-5404). IDO is also expressed in myeloid-derived suppressor cells (MDSC) in cancer tissues, and MDSC also produces kynurenine (Yu et al. (J. Immunol. (2013) 190, 3783-3797)). 5 [Compound 1] o nh2 Kynurenine is known to suppress the same type of T cell response (Frumento et al. (J. Exp. Med. (2002) 196, 459-468); and a mechanism has been suggested, in which tumor cells evade antitumor immune responses through such inhibition, and proliferation of glioma cells is 20 promoted through an autocrine proliferation mechanism in which kynurenine acts as an endogenous ligand for the aryl hydrocarbon receptor expressed on gliomas (Optiz et al. (mentioned above)). Kynurenine is converted to anthranilic acid (shown as Compound 2) by kynurenidase, and to 3-hydroxykynurenine (shown as Compound 3) by kynurenine 3-hydroxylase. Anthranilic acid and 3-hydroxykynurenine are both converted to 25 3-hydroxyanthranilic acid, the precursor of NAD. [Compound 2] 2023229507  12 Sep 2023 [Compound 3] 5 Kynurenine is converted to kynurenic acid (shown as Compound 4) by kynurenine aminotransferase. Examples of a non-limiting embodiment of cancer tissue-specific compounds, particularly cancer cell-specific metabolites, used in the present invention preferably include such amino acid metabolites such as kynurenine and its metabolites such as anthranilic acid, 3-hydroxykynurenine, and kynurenic acid. 0 [Compound 4] (4) Arachidonic acid metabolites such as prostaglandin E2 15 Prostaglandin E2 (PGE2) (Compound 5) is an arachidonic acid metabolite called a prostanoid, which includes thromboxane and prostaglandin synthesized by cyclooxygenase (COX)-1 / 2 (Warner and Mitchell (FASEB J. (2004) 18, 790-804)). PGE2 promotes the proliferation of colon cancer cells and suppresses their apoptosis (Sheng et al. (Cancer Res. (1998) 58, 362-366)). Cyclooxygenase expression is known to be altered in many cancer cells. 20 More specifically, while COX-1 is expressed constitutively in almost all tissues, COX-2 has been found to be mainly induced by certain types of inflammatory cytokines and cancer genes in tumors (Warner and Mitchell (mentioned above)). In addition, COX-2 overexpression has been reported to be related to bad prognosis for breast cancer (Denkert et al. (Clin. Breast Cancer 2023229507  12 Sep 2023 (2004) 4, 428-433)), and rapid disease progression for ovarian cancer (Denker et al. (Mod. Pathol. (2006) 19, 1261-1269)). Inhibitory T cells that have infiltrated into cancer tissues also produce prostaglandin E2 (Curr. Med. Chem. (2011) 18, 5217-5223). Small molecules such as the arachidonic acid metabolites prostaglandin and leukotriene are known to act as a stimulating 5 factor that regulates autocrine and / or paracrine growth of cancer (Nat. Rev. Cancer (2012) 12 (11) 782-792). Examples of a non-limiting embodiment of cancer tissue-specific compounds used in the present invention, particularly cancer cell-specific metabolites and immune cell-specific metabolites that have infiltrated into cancer tissues, preferably include such arachidonic acid metabolites such as prostaglandin E2. Besides prostaglandin E2, production 0 of thromboxane A2 (TXA2) is enhanced in cancer tissues such as colorectal cancer tissues (J. Lab. Clin. Med. (1993) 122, 518-523), and thromboxane A2 can be suitably presented as a non-limiting embodiment of an arachidonic acid metabolite of the present invention. [Compound 5] (5) Nucleosides carrying a purine ring structure such as adenosine, adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP) When cancer cells undergo cell death, a large amount of ATP in the cell is known to leak out to the outside of the cells. Therefore, the ATP concentration is remarkably higher in cancer 20 tissues than in normal tissues (PLoS One. (2008) 3, e2599). Multiple types of cells release adenine nucleotides in the form of ATP, ADP, and AMP. Metabolism takes place through an extracellular enzyme on the cell surface such as extracellular 5'-nucleotidase (ecto-5'-nucleotidase) (CD73) (Resta and Thompson (Immunol. Rev. (1998) 161, 95-109) and Sadej et al. (Melanoma Res. (2006) 16, 213-222)). Adenosine is a purine nucleoside that exists 25 constitutively at low concentration in the extracellular environment, but in hypoxic tissues found in solid cancers, a remarkable increase in the extracellular adenosine concentration has been reported (Blay and Hoskin (Cancer Res. (1997) 57, 2602-2605). CD73 is expressed on the surface of immune cells and tumors (Kobie et al. (J. Immunol. (2006) 177, 6780-6786)), and its 2023229507  12 Sep 2023 activity has been found to be increased in breast cancer (Canbolat et al. (Breast Cancer Res. Treat. (1996) 37, 189-193)), stomach cancer (Durak et al. (Cancer Lett. (1994) 84, 199-202)), pancreatic cancer (Flocke and Mannherz (Biochim. Biophys. Acta (1991) 1076, 273-281), and glioblastoma (Bardot et al. (Br. J. Cancer (1994) 70, 212-218)). It has been proposed that the 5 accumulation of adenosine in cancer tissues may be caused by an increase in the intracellular adenosine production through dephosphorylation of AMP by 5'-nucleotidase in the cytoplasm (Headrick and Willis (Biochem. J. (1989) 261, 541-550)). Furthermore, inhibitory T cells and such that have infiltrated into cancer tissues also express ATPase and produce adenosine (Proc. Natl. Acad. Sci. (2006) 103 (35), 13132-13137; Curr. Med. Chem. (2011) 18, 5217-5223). The 0 produced adenosine is considered to be rendering the cancer tissue an immunosuppressive environment through adenosine receptors such as the A2A receptor (Curr. Med. Chem. (2011),18, 5217-23). Examples of a non-limiting embodiment of the cancer tissue-specific compound used in the present invention preferably include ATP, ADP, AMP, and adenosine which accumulate at high concentration in cancer tissues through such metabolism of purine 5 nucleotides such as ATP. Furthermore, since adenosine is degraded to inosine by adenosine deaminase, inosine accumulates at high concentration. (6) Uric acid Uric acid is a product of the metabolic pathway of purine nucleosides in vivo, and is 0 released to the outside of cells such as the interstitial space and blood. In recent years, it has been found to be released from dead cells that are present at sites of lesions such as cancer tissues (Nat. Med. (2007) 13, 851-856). Examples of a non-limiting embodiment of cancer tissue-specific compounds used in the present invention preferably include such uric acid which accumulates at high concentration in cancer tissues due to metabolism of purine nucleotides such 25 as ATP. (7) 1-Methyl nicotinamide The enzyme nicotinamide N-methyl transferase is known to be highly expressed in several human cancer tissues. When this enzyme produces the stable metabolite 30   1-methylnicotinamide from nicotinamide, the methyl group of S-adenosylmethionine (SAM) which serves as a methyl donor is consumed; therefore, the high expression of nicotinamide N-methyltransferase has been suggested to contribute to tumorigenesis through a mechanism that impairs the DNA methylation ability accompanying a decrease in the SAM concentration in cancer cells (Ulanovskaya et al. (Nat. Chem. Biol. (2013) 9 (5) 300-306)). The stable 35 metabolite of this enzyme, 1-methylnicotinamide is known to be secreted to the outside of cancer cells (Yamada et al. (J. Nutr. Sci. Vitaminol. (2010) 56, 83-86)), and preferable examples of a 2023229507  12 Sep 2023 non-limiting embodiment of cancer tissue-specific compounds used in the present invention include 1-methylnicotinamide and such which accumulate at high concentration in cancer tissues through nicotinamide metabolism. 5   Inflammatory tissue-specific compounds The term "compound specific to inflammatory tissue (inflammatory tissue-specific compound)" as used herein refers to a compound that is present differentially in inflammatory tissues as compared to non-inflammatory tissues. Herein, suitable examples of "inflammatory tissues" include: 0 joints with rheumatoid arthritis or osteoarthritis; lungs (alveoli) with bronchial asthma or COPD; digestive organs of inflammatory bowel disease, Crohn's disease, or ulcerative colitis; fibrotic tissues of fibrosis of the liver, kidney, or lung; tissues undergoing rejection reaction in organ transplantation; 5 blood vessels and heart (myocardium) in arteriosclerosis or heart failure; visceral fat in metabolic syndrome; skin tissues in atopic detrmatitis or other dermatitis; and spinal nerves in disk herniation or chronic low back pain. 0 Inflammatory tissue-specific metabolites “Inflammatory tissue-specific metabolite” refers to metabolites highly produced by immune cells that have infiltrated into inflammatory tissues, and metabolites highly produced by specifically normal cells that have been damaged in inflammatory tissues. Examples of infiltrating immune cells include effector T cells, mature dendritic cells, neutrophils, granule 25 cells (mast cells), and basophils. Furthermore, metabolites in the present invention include compounds that are released from inside the cells to the outside of the cells when the cells that are present in inflammatory tissues (immune cells and normal cells) die by apoptosis, necrosis, or such. Examples of a non-limiting embodiment of the inflammatory tissue-specific compounds 30 or inflammatory tissue-specific metabolites used in the present invention preferably include at least one compound selected from the compounds below. At least one compound means including cases where the antigen-binding activity of a same antigen-binding domain described below depends on one type of inflammatory tissue-specific compound or metabolite, as well as cases where it depends on several types of inflammatory tissue-specific compounds or 35 metabolites. 2023229507  12 Sep 2023 (1) Arachidonic acid metabolites such as prostaglandin E2 The PGE2 concentration has been known to be high in rheumatoid arthritis and osteoarthritis (Eur. J. Clin. Pharmacol. (1994) 46, 3-7.; Clin. Exp. Rheumatol. (1999) 17, 151-160; Am. J. Vet. Res. (2004) 65, 1269-1275). Examples of a non-limiting embodiment of 5 inflamatory tissue-specific compounds, particularly inflammatory tissue-specific metabolites and metabolites specific to immune cells that infiltrate into inflammatory tissues used in the present invention preferably include such arachidonic acid metabolites such as prostaglandin E2. (2) Nucleosides carrying a purine ring structure such as adenosine, adenosine triphosphate (ATP), 0 adenosine diphosphate (ADP), and adenosine monophosphate (AMP) ATP concentration is known to be high in pulmonary alveoli where inflammation caused by bronchial asthma is taking place (Nat. Med. (2007) 13, 913-919). ATP concentration is also known to be high in pulmonary alveoli where inflammation caused by COPD is taking place (Am. J. Respir. Crit. Care Med. (2010) 181, 928-934). Furthermore, adenosine concentration 5 has been observed to be high in the joint fluid of rheumatoid arthritis patients (Journal of Pharmaceutical and Biomedical Analysis (2004) 36, 877-882). Furthermore, ATP concentration is known to be high in tissues where a rejection reaction is taking place due to GVHD (Nat. Med. (2010) 16, 1434-1438). Adenosine concentration is known to be enhanced in fibrotic tissues of the liver, kidney, and lung (FASEB J. (2008) 22, 2263-2272; J. Immunol. (2006) 176, 0 4449-4458; J. Am. Soc. Nephrol. (2011) 22 (5), 890-901; PLoS ONE J. (2010) 5 (2), e9242). Furthermore, ATP concentration has been observed to be increased in fibrotic tissues of pulmonary fibrosis patients (Am. J. Respir. Crit. Care Med. (2010) 182, 774-783). Examples of a non-limiting embodiment of an inflammatory tissue-specific compound used in the present invention suitably include ATP, ADP, AMP, adenosine and such which accumulate at high 25 concentration in inflammatory tissues by metabolism of such purine nucleotides such as ATP. In addition, inosine accumulates at a high concentration due to degradation of adenosine by adenosine deaminase to produce inosine. (3) Uric acid 30 Uric acid is a product of the metabolic pathway of purine nucleosides in vivo, and is released to the outside of cells such as the interstitial space and blood. In recent years, uric acid released from cells undergoing necrosis has been found to promote inflammatory response (J. Clin. Invest. (2010) 120 (6), 1939-1949). Examples of a non-limiting embodiment of inflammatory tissue-specific compounds to be used in the present invention suitably include such 35 uric acid which accumulates at high concentration in inflammatory tissues due to metabolism of purine nucleotides such as ATP. 2023229507  12 Sep 2023 Antigen-binding domain Herein, an “antigen-binding domain” may be of any structure as long as it binds to an antigen of interest. Such domains preferably include, for example: 5 antibody heavy-chain and light-chain variable regions; a module of about 35 amino acids called A domain which is contained in the in vivo cell membrane protein Avimer (International Publication No. WO 2004 / 044011, International Publication No. WO 2005 / 040229); Adnectin containing the 10Fn3 domain which binds to the protein moiety of fibronectin, a 0 glycoprotein expressed on cell membrane (International Publication No. WO 2002 / 032925); Affibody which is composed of a 58-amino acid three-helix bundle based on the scaffold of the IgG-binding domain of Protein A (International Publication No. WO 1995 / 001937); Designed Ankyrin Repeat proteins (DARPins) which are a region exposed on the molecular surface of ankyrin repeats (AR) having a structure in which a subunit consisting of a turn 5 comprising 33 amino acid residues, two antiparallel helices, and a loop is repeatedly stacked (International Publication No. WO 2002 / 020565); Anticalins and such, which are domains consisting of four loops that support one side of a barrel structure composed of eight circularly arranged antiparallel strands that are highly conserved among lipocalin molecules such as neutrophil gelatinase-associated lipocalin (NGAL) 0 (International Publication No. WO 2003 / 029462); and the concave region formed by the parallel-sheet structure inside the horseshoe-shaped structure constituted by stacked repeats of the leucine-rich-repeat (LRR) module of the variable lymphocyte receptor (VLR) which does not have the immunoglobulin structure and is used in the system of acquired immunity in jawless vertebrate such as lampery and hagfish (International 25 Publication No. WO 2008 / 016854). Preferred antigen-binding domains of the present invention include, for example, those having antibody heavy-chain and light-chain variable regions. Preferred examples of antigen-binding domains include “single chain Fv (scFv)”, “single chain antibody”, “Fv”, “single chain Fv 2 (scFv2)”, “Fab”, and “F(ab’)2”. The antigen-binding domains of antigen-binding molecules of the present invention can 30 bind to an identical epitope. Such identical epitope can be present, for example, in a protein comprising the amino acid sequence of SEQ ID NO: 1. Alternatively, each of the antigen-binding domains of antigen-binding molecules of the present invention can bind to a different epitope. Herein, the different epitope can be present in, for example, a protein comprising the amino acid sequence of SEQ ID NO: 1. 35 Specificity 2023229507  12 Sep 2023 “Specific” means that one of the molecules that specifically bind does not substantially bind to molecules other than the single or plurality of partner molecules it binds to. Furthermore, “specific” is also used when an antigen-binding domain is specific to a particular epitope among multiple epitopes in an antigen. When an epitope bound by an antigen-binding 5 domain is contained in multiple different antigens, antigen-binding molecules containing the antigen-binding domain can bind to various antigens that have the epitope. Here, “does not substantially bind” is determined according to the method described in the above-mentioned section on binding activity, and refers to the binding activity of a molecule that specifically binds to a molecule other than the partner molecule, where the binding activity is not more than 80%, 0 normally not more than 50%, preferably not more than 30%, or particularly preferably not more than 15% of the binding activity to its partner molecule. Cytotoxic activity In a non-limiting embodiment, the present invention provides antigen-binding 5 molecules that comprise an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a cancer-tissue specific compound, and which have cytotoxic activity against cells expressing a membrane-type molecule on their cell membrane; and pharmaceutical compositions comprising these antigen-binding molecules as an active ingredient. In the present invention, cytotoxic activity includes, for example, antibody-dependent 0 cell-mediated cytotoxicity (ADCC) activity, complement-dependent cytotoxicity (CDC) activity, and cytotoxic activity by T cells. In the present invention, CDC activity refers to cytotoxic activity by the complement system. On the other hand, ADCC activity refers to the activity of immune cells to damage target cells when the immune cells and such bind to the Fc region of antigen-binding molecules comprising an antigen-binding domain that binds to a membrane-type 25 molecule expressed on the cell membrane of target cells via an Fcy receptor expressed on the immune cells. Whether an antigen-binding molecule of interest has an ADCC activity or whether it has a CDC activity can be determined using known methods (for example, Current Protocols in Immunology, Chapter 7. Immunologic studies in humans, Editor, Coligan et al., (1993)). Specifically, effector cells, complement solution, and target cells are first prepared. 30 (1) Preparation of effector cells Spleen is removed from a CBA / N mouse or the like, and spleen cells are dispersed in an RPMI1640 medium (Invitrogen). After the cells are washed in the same medium containing 10% fetal bovine serum (FBS, HyClone), effector cells are prepared by adjusting the spleen cell concentration to 5 x 106 / mL. 35 (2) Preparation of complement solution Baby Rabbit Complement (CEDARLANE) is diluted 10-fold in a culture medium 2023229507  12 Sep 2023 (Invitrogen) containing 10% FBS to prepare a complement solution. (3) Preparation of target cells The target cells can be radioactively labeled by culturing cells expressing the antigen with 0.2 mCi of 51Cr-sodium chromate- (GE Healthcare Bio-Sciences) in a DMEM medium 5 containing 10% FBS for one hour at 37°C. After radioactive labeling, cells are washed three times in an RPMI1640 medium containing 10% FBS, and the target cells can be prepared by adjusting the cell concentration to 2 x 105 / mL. ADCC activity or CDC activity can be measured by the method described below. In the case of ADCC activity measurement, 50 pL each of the target cell and antigen-binding 0 molecule are added to a 96-well U-bottom plate (Becton Dickinson), and allowed to react for 15 minutes at room temperature. Then, 100 pL of effector cells are added to the plate and this plate is placed in a carbon dioxide incubator for four hours. The final concentration of the antigen-binding molecule may be set, for example, to 0 pg / mL or 10 pg / mL. After incubation, 100 pL of the supernatant is collected from each well, and the radioactivity is measured with a 5 gamma counter (COBRAII AUTO-GAMMA, MODEL D5005, Packard Instrument Company). The cytotoxic activity (%) can be calculated using the measured values according to the equation: (A - C) / (B - C) x 100. A represents the radioactivity (cpm) in each sample, B represents the radioactivity (cpm) in a sample to which 1% NP-40 (Nacalai Tesque) has been added, and C represents the radioactivity (cpm) of a sample containing the target cells alone. 0 Meanwhile, in the case of CDC activity measurement, 50 pL of target cell and 50 pL of an antigen-binding molecule are added to a 96-well flat-bottomed plate (Becton Dickinson), and allowed to react for 15 minutes on ice. Then, 100 pL of a complement solution is added to the plate, and this plate is placed in a carbon dioxide incubator for four hours. The final concentration of the antigen-binding molecule may be set, for example, to 0 pg / mL or 3 pg / mL. 25 After incubation, 100 pL of supernatant is collected from each well, and the radioactivity is measured with a gamma counter. The cytotoxic activity can be calculated in the same way as in the determination of ADCC activity. The later-described modified antigen-binding molecules to which cytotoxic substances such as chemotherapeutic agents, toxic peptides, or radioactive chemical substances have been 30 ligated can also be suitably used as the antigen-binding molecules of the present invention having cytotoxic activity. Such modified antigen-binding molecules (hereinafter referred to as “antigen-binding molecule-drug conjugate”) can be obtained by chemically modifying the obtained antigen-binding molecules. Methods that have been already established in the field of antibody-drug conjugates and such may be used appropriately as a method for modifying 35 antigen-binding molecules. Furthermore, a modified antigen-binding molecule with a linked toxic peptide can be obtained by expressing in an appropriate host cell a fusion gene produced by 2023229507  12 Sep 2023 linking a gene encoding the toxic peptide in frame with a gene encoding an antigen-binding molecule of the present invention, and then isolating the molecule from the culture solution of the cells. 5 Neutralizing activity The present invention provides in a non-limiting embodiment a pharmaceutical composition that induces an immune response, comprising as an active ingredient an antigen-binding molecule that contains an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a cancer tissue-specific compound and has a 0 neutralizing activity against a membrane-type molecule. In another non-limiting embodiment, the present invention provides a pharmaceutical composition that induces an immune response, comprising as an active ingredient an antigen-binding molecule that contains an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a cancer tissue-specific compound and has a neutralizing activity against a membrane-type molecule in 5   addition to a cytotoxic activity against cells expressing the membrane-type molecule on their cell membrane. Generally, a neutralizing activity refers to an activity of inhibiting the biological activity of a ligand which has a biological activity towards cells, such as viruses and toxins. Thus, a substance having a neutralizing activity refers to a substance that binds to a ligand or a receptor to which the ligand binds and inhibits the binding between the ligand and the receptor. 0 A receptor whose binding to the ligand has been blocked by the neutralizing activity will not be able to exhibit the biological activity through the receptor. When the antigen-binding molecule is an antibody, the antibody having such a neutralizing activity is generally called a neutralizing antibody. The neutralizing activity of a test substance may be measured by comparing the biological activities in the presence of a ligand between conditions when the test substance is 25 present or absent. A suitable example of a major ligand for the IL-6 receptor is IL-6, which is shown in SEQ ID NO: 27. The IL-6 receptor, which is an I-type membrane protein whose amino terminus forms the extracellular domain, forms a hetero-tetramer with the gp130 receptor which was induced by IL-6 to dimerize (Heinrich et al. (Biochem. J. (1998) 334, 297-314)). 30 Formation of the heterotetramer activates Jak associated with the gp130 receptor. Jak carries out autophosphorylation and receptor phosphorylation. The phosphorylation sites of the receptor and of Jak serve as binding sites for molecules belonging to the Stat family having SH2 such as Stat3, and for the MAP kinases, PI3 / Akt, and other proteins and adapters having SH2. Next, Stat that bound to the gp130 receptor is phosphorylated by Jak. The phosphorylated Stat 35 dimerizes and translocates to the nucleus, and regulates transcription of target genes. Jak and Stat can also be involved in the signaling cascade through receptors of other classes. A 2023229507  12 Sep 2023 deregulated IL-6 signaling cascade is observed in inflammation and pathological conditions of autoimmune diseases, and cancers such as prostate cancer and multiple myeloma. Stat3 which may act as an oncogene is constitutively activated in many cancers. In prostate cancer and multiple myeloma, there is a crosstalk between the signaling cascade from the IL-6 receptor and 5 the signaling cascade from members of the epidermal growth factor receptor (EGFR) family (Ishikawa et al. (J. Clin. Exp. Hematopathol. (2006) 46 (2), 55-66)). Such intracellular signaling cascades are different for each cell type; therefore, an appropriate target molecule can be set according to each of the target cells of interest, and the target molecule is not limited to the above-mentioned factors. The neutralization activity can 0 be evaluated by measuring the in vivo signal activation. Furthermore, activation of in vivo signals can also be detected by using as an indicator the transcription-inducing action on a target gene that exists downstream of the in vivo signaling cascade. A change in the transcription activity of a target gene can be detected by the principle of a reporter assay. Specifically, a reporter gene such as the green fluorescence protein (GFP) or luciferase is placed downstream of 5 a transcription factor or a promoter region of the target gene; and a change in transcription activity can be measured in terms of reporter activity by measuring the reporter activity. Commercially available kits for measuring in vivo signal activation can be suitably used (for example, the Mercury Pathway Profiling Luciferase System (Clontech)). Furthermore, as a method for measuring the neutralization activity on a receptor ligand 0 in the EGF receptor family and such which acts on a signaling cascade that typically works toward enhancing cell proliferation, neutralization activity of an antigen-binding molecule can be evaluated by measuring the proliferation activity of the target cells. For example, the following method is suitably used as a method for measuring or evaluating inhibitory effects based on the neutralization activity of an anti-HB-EGF antibody against the proliferation of cells whose 25 proliferation is promoted by EGF family growth factors such as HB-EGF. As a method for evaluating or measuring the activity of inhibiting cell proliferation in a test tube, a method that measures the incorporation by living cells of [3H]-labeled thymidine added to the culture medium as an index of the DNA replication ability is used. As a more convenient method, a dye exclusion method that measures under a microscope the ability of a cell to release a dye such 30 as trypan blue to the outside of the cell, or the MTT method is used. The latter makes use of the ability of living cells to convert 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), which is a tetrazolium salt, to a blue formazan product. More specifically, a test antibody is added along with a ligand to the culture solution of a test cell; and after a certain period of time has elapsed, an MTT solution is added to the culture, and this is left to stand for a 35 certain amount of time to let the cell incorporate MTT. As a result, MTT which is a yellow compound is converted to a blue compound by succinate dehydrogenase in the mitochondria of 2023229507  12 Sep 2023 the cell. After this blue product is dissolved for coloration, its absorbance is measured and used as an indicator of the number of viable cells. Besides MTT, reagents such as MTS, XTT, WST-1, and WST-8 are also commercially available (Nacalai Tesque, and such), and can be suitably used. For measurement of the activity, a binding antibody that has the same isotype as 5 the anti-HB-EGF antibody but does not have the cell proliferation-inhibiting activity can be used as a control antibody in the same manner as the anti-HB-EGF antibody, and the anti-HB-EGF antibody is judged to have the activity when it shows a stronger cell proliferation-inhibiting activity than the control antibody. As cells for evaluating activity, for example, cells showing HB-EGF-promoted 0 proliferation such as the RMG-1 cell line which is an ovarian cancer cell line may be suitably used; and mouse Ba / F3 cells transformed with a vector in which a gene encoding hEGFR / mG-CSFR, which is a fusion protein of the extracellular domain of human EGFR fused in frame with the intracellular domain of the mouse G-CSF receptor, is linked so as to allow expression, may also be suitably used. This way, those skilled in the art may appropriately 5   select cells for evaluating activity to measure the cell proliferation activity mentioned above. Antibody Herein, “antibody” refers to a natural immunoglobulin or an immunoglobulin produced by partial or complete synthesis. Antibodies can be isolated from natural sources such as 0 naturally-occurring plasma and serum, or culture supernatants of antibody-producing hybridomas. Alternatively, antibodies can be partially or completely synthesized using techniques such as genetic recombination. Preferred antibodies include, for example, antibodies of an immunoglobulin isotype or subclass belonging thereto. Known human immunoglobulins include antibodies of the following nine classes (isotypes): IgG1, IgG2, IgG3, 25 IgG4, IgA1, IgA2, IgD, IgE, and IgM. Of these isotypes, antibodies of the present invention include IgG1, IgG2, IgG3, and IgG4. A number of allotype sequences of human IgG1, human IgG2, human IgG3, and human IgG4 constant regions due to gene polymorphisms are described in “Sequences of proteins of immunological interest”, NIH Publication No. 91-3242. Any of such sequences may be used in the present invention. In particular, for the human 30 IgG1sequence, the amino acid sequence at positions 356 to 358 as indicated by EU numbering may be DEL or EEM. Several allotype sequences due to genetic polymorphisms have been described in “Sequences of proteins of immunological interest”, NIH Publication No. 91-3242 for the human IgK (Kappa) constant region and human IgA (Lambda) constant region, and any of the sequences may be used in the present invention. 35 Methods for producing an antibody with desired binding activity are known to those skilled in the art. Below is an example that describes a method for producing an antibody that 2023229507  12 Sep 2023 binds to IL-6R (anti-IL-6R antibody). Antibodies that bind to an antigen other than IL-6R can also be produced according to the example described below. Anti-IL-6R antibodies can be obtained as polyclonal or monoclonal antibodies using known methods. The anti-IL-6R antibodies preferably produced are monoclonal antibodies 5 derived from mammals. Such mammal-derived monoclonal antibodies include antibodies produced by hybridomas or host cells transformed with an expression vector carrying an antibody gene by genetic engineering techniques. “Humanized antibodies” or “chimeric antibodies” are included in the monoclonal antibodies of the present invention. Monoclonal antibody-producing hybridomas can be produced using known techniques, 0 for example, as described below. Specifically, mammals are immunized by conventional immunization methods using an IL-6R protein as a sensitizing antigen. Resulting immune cells are fused with known parental cells by conventional cell fusion methods. Then, hybridomas producing an anti-IL-6R antibody can be selected by screening for monoclonal antibody-producing cells using conventional screening methods. 5 Specifically, monoclonal antibodies are prepared as mentioned below. First, the IL-6R gene whose nucleotide sequence is disclosed in SEQ ID NO: 2 can be expressed to produce an IL-6R protein shown in SEQ ID NO: 1, which will be used as a sensitizing antigen for antibody preparation. That is, a gene sequence encoding IL-6R is inserted into a known expression vector, and appropriate host cells are transformed with this vector. The desired human IL-6R 0 protein is purified from the host cells or their culture supernatants by known methods. In order to obtain soluble IL-6R from culture supernatants, for example, a protein consisting of the amino acids at positions 1 to 357 in the IL-6R polypeptide sequence of SEQ ID NO: 1, such as described in Mullberg et al. (J. Immunol. (1994) 152 (10), 4958-4968), is expressed as a soluble IL-6R, instead of the IL-6R protein of SEQ ID NO: 1. Purified natural IL-6R protein can also 25 be used as a sensitizing antigen. The purified IL-6R protein can be used as a sensitizing antigen for immunization of mammals. A partial IL-6R peptide may also be used as a sensitizing antigen. In this case, a partial peptide can be prepared by chemical synthesis based on the amino acid sequence of human IL-6R, or by inserting a partial IL-6R gene into an expression vector for expression. 30 Alternatively, a partial peptide can be produced by degrading an IL-6R protein with a protease. The length and region of the partial IL-6R peptide are not limited to particular embodiments. A preferred region can be arbitrarily selected from the amino acid sequence at amino acid positions 20 to 357 in the amino acid sequence of SEQ ID NO: 1. The number of amino acids forming a peptide to be used as a sensitizing antigen is preferably at least five or more, six or more, or 35 seven or more. More specifically, a peptide of 8 to 50 residues, more preferably 10 to 30 residues can be used as a sensitizing antigen. 2023229507  12 Sep 2023 For sensitizing antigen, alternatively it is possible to use a fusion protein prepared by fusing a desired partial polypeptide or peptide of the IL-6R protein with a different polypeptide. For example, antibody Fc fragments and peptide tags are preferably used to produce fusion proteins to be used as sensitizing antigens. Vectors for expression of such fusion proteins can 5 be constructed by fusing in frame genes encoding two or more desired polypeptide fragments and inserting the fusion gene into an expression vector as described above. Methods for producing fusion proteins are described in Molecular Cloning 2nd ed. (Sambrook, J et al., Molecular Cloning 2nd ed., 9.47-9.58 (1989) Cold Spring Harbor Lab. Press). Methods for preparing IL-6R to be used as a sensitizing antigen, and immunization methods using IL-6R are 0 specifically described in WO 2003 / 000883, WO 2004 / 022754, WO 2006 / 006693, and such. There is no particular limitation on the mammals to be immunized with the sensitizing antigen. However, it is preferable to select the mammals by considering their compatibility with the parent cells to be used for cell fusion. In general, rodents such as mice, rats, and hamsters, rabbits, and monkeys are preferably used. 5 The above animals are immunized with a sensitizing antigen by known methods. Generally performed immunization methods include, for example, intraperitoneal or subcutaneous injection administration of a sensitizing antigen into mammals. Specifically, a sensitizing antigen is appropriately diluted with PBS (Phosphate-Buffered Saline), physiological saline, or the like. If desired, a conventional adjuvant such as Freund’s complete adjuvant is 0   mixed with the antigen, and the mixture is emulsified. Then, the sensitizing antigen is administered to a mammal several times at 4- to 21-day intervals. Appropriate carriers may be used in immunization with the sensitizing antigen. In particular, when a low-molecular-weight partial peptide is used as the sensitizing antigen, it is sometimes desirable to couple the sensitizing antigen peptide to a carrier protein such as albumin or keyhole limpet hemocyanin for 25 immunization. Alternatively, hybridomas producing a desired antibody can be prepared using DNA immunization as mentioned below. DNA immunization is an immunization method that confers immunostimulation by expressing a sensitizing antigen in an animal immunized as a result of administering a vector DNA constructed to allow expression of an antigen 30 protein-encoding gene in the animal. As compared to conventional immunization methods in which a protein antigen is administered to animals to be immunized, DNA immunization is expected to be superior in that: - immunostimulation can be provided while retaining the structure of a membrane protein such as IL-6R; and 35 - there is no need to purify the antigen for immunization. In order to prepare a monoclonal antibody of the present invention using DNA 2023229507  12 Sep 2023 immunization, first, a DNA expressing an IL-6R protein is administered to an animal to be immunized. The IL-6R-encoding DNA can be synthesized by known methods such as PCR. The obtained DNA is inserted into an appropriate expression vector, and then this is administered to an animal to be immunized. Preferably used expression vectors include, for example, 5 commercially-available expression vectors such as pcDNA3.1. Vectors can be administered to an organism using conventional methods. For example, DNA immunization is performed by using a gene gun to introduce expression vector-coated gold particles into cells in the body of an animal to be immunized. Antibodies that recognized IL-6R can also be produced by the methods described in WO 2003 / 104453. 0 After immunizing a mammal as described above, an increase in the titer of an IL-6R-binding antibody is confirmed in the serum. Then, immune cells are collected from the mammal, and then subjected to cell fusion. In particular, splenocytes are preferably used as immune cells. A mammalian myeloma cell is used as a cell to be fused with the above-mentioned 5 immune cells. The myeloma cells preferably comprise a suitable selection marker for screening. A selection marker confers characteristics to cells for their survival (or death) under a specific culture condition. Hypoxanthine-guanine phosphoribosyltransferase deficiency (hereinafter abbreviated as HGPRT deficiency) and thymidine kinase deficiency (hereinafter abbreviated as TK deficiency) are known as selection markers. Cells with HGPRT or TK deficiency have 0 hypoxanthine-aminopterin-thymidine sensitivity (hereinafter abbreviated as HAT sensitivity). HAT-sensitive cells cannot synthesize DNA in a HAT selection medium, and are thus killed. However, when the cells are fused with normal cells, they can continue DNA synthesis using the salvage pathway of the normal cells, and therefore they can grow even in the HAT selection medium. 25 HGPRT-deficient and TK-deficient cells can be selected in a medium containing 6-thioguanine, 8-azaguanine (hereinafter abbreviated as 8AG), or 5’-bromodeoxyuridine, respectively. Normal cells are killed because they incorporate these pyrimidine analogs into their DNA. Meanwhile, cells that are deficient in these enzymes can survive in the selection medium, since they cannot incorporate these pyrimidine analogs. In addition, a selection 30 marker referred to as G418 resistance provided by the neomycin-resistant gene confers resistance to 2-deoxystreptamine antibiotics (gentamycin analogs). Various types of myeloma cells that are suitable for cell fusion are known. For example, myeloma cells including the following cells can be preferably used: P3(P3x63Ag8.653) (J. Immunol. (1979) 123 (4), 1548-1550); 35 P3x63Ag8U.1 (Current Topics in Microbiology and Immunology (1978)81, 1-7); NS-1 (C. Eur. J. Immunol. (1976)6 (7), 511-519); 2023229507  12 Sep 2023 MPC-11 (Cell (1976) 8 (3), 405-415); SP2 / 0 (Nature (1978) 276 (5685), 269-270); FO (J. Immunol. Methods (1980) 35 (1-2), 1-21); S194 / 5.XX0.BU.1 (J. Exp. Med. (1978) 148 (1), 313-323); 5 R210 (Nature (1979) 277 (5692), 131-133), etc. Cell fusions between the immunocytes and myeloma cells are essentially carried out using known methods, for example, a method by Kohler and Milstein et al. (Methods Enzymol. (1981) 73: 3-46). More specifically, cell fusion can be carried out, for example, in a conventional culture 0 medium in the presence of a cell fusion-promoting agent. The fusion-promoting agents include, for example, polyethylene glycol (PEG) and Sendai virus (HVJ). If required, an auxiliary substance such as dimethyl sulfoxide is also added to improve fusion efficiency. The ratio of immune cells to myeloma cells may be determined at one’s own discretion, preferably, for example, one myeloma cell for every one to ten immunocytes. Culture media to 5 be used for cell fusions include, for example, media that are suitable for the growth of myeloma cell lines, such as RPMI1640 medium and MEM medium, and other conventional culture medium used for this type of cell culture. In addition, serum supplements such as fetal calf serum (FCS) may be preferably added to the culture medium. For cell fusion, predetermined amounts of the above immune cells and myeloma cells 0 are mixed well in the above culture medium. Then, a PEG solution (for example, the average molecular weight is about 1,000 to 6,000) prewarmed to about 37°C is added thereto at a concentration of generally 30% to 60% (w / v). This is gently mixed to produce desired fusion cells (hybridomas). Then, an appropriate culture medium mentioned above is gradually added to the cells, and this is repeatedly centrifuged to remove the supernatant. Thus, cell fusion 25 agents and such which are unfavorable to hybridoma growth can be removed. The hybridomas thus obtained can be selected by culture using a conventional selective medium, for example, HAT medium (a culture medium containing hypoxanthine, aminopterin, and thymidine). Cells other than the desired hybridomas (non-fused cells) can be killed by continuing culture in the above HAT medium for a sufficient period of time. Typically, the 30 period is several days to several weeks. Then, hybridomas producing the desired antibody are screened and singly cloned by conventional limiting dilution methods. The hybridomas thus obtained can be selected using a selection medium based on the selection marker possessed by the myeloma used for cell fusion. For example, HGPRT- or TK-deficient cells can be selected by culture using the HAT medium (a culture medium 35 containing hypoxanthine, aminopterin, and thymidine). Specifically, when HAT-sensitive myeloma cells are used for cell fusion, cells successfully fused with normal cells can selectively 2023229507  12 Sep 2023 proliferate in the HAT medium. Cells other than the desired hybridomas (non-fused cells) can be killed by continuing culture in the above HAT medium for a sufficient period of time. Specifically, desired hybridomas can be selected by culture for generally several days to several weeks. Then, hybridomas producing the desired antibody are screened and singly cloned by 5 conventional limiting dilution methods. Desired antibodies can be preferably selected and singly cloned by screening methods based on known antigen / antibody reaction. For example, an IL-6R-binding monoclonal antibody can bind to IL-6R expressed on the cell surface. Such a monoclonal antibody can be screened by fluorescence activated cell sorting (FACS). FACS is a system that assesses the 0 binding of an antibody to cell surface by analyzing cells contacted with a fluorescent antibody using laser beam, and measuring the fluorescence emitted from individual cells. To screen for hybridomas that produce a monoclonal antibody of the present invention by FACS, IL-6R-expressing cells are first prepared. Cells preferably used for screening are mammalian cells in which IL-6R is forcedly expressed. As control, the activity of an antibody 5 to bind to cell-surface IL-6R can be selectively detected using non-transformed mammalian cells as host cells. Specifically, hybridomas producing an anti-IL-6R monoclonal antibody can be isolated by selecting hybridomas that produce an antibody which binds to cells forced to express IL-6R, but not to host cells. Alternatively, the activity of an antibody to bind to immobilized IL-6R-expressing cells 0 can be assessed based on the principle of ELISA. For example, IL-6R-expressing cells are immobilized to the wells of an ELISA plate. Culture supernatants of hybridomas are contacted with the immobilized cells in the wells, and antibodies that bind to the immobilized cells are detected. When the monoclonal antibodies are derived from mouse, antibodies bound to the cells can be detected using an anti-mouse immunoglobulin antibody. Hybridomas producing a 25 desired antibody having the antigen-binding ability are selected by the above screening, and they can be cloned by a limiting dilution method or the like. Monoclonal antibody-producing hybridomas thus prepared can be passaged in a conventional culture medium, and stored in liquid nitrogen for a long period. The above hybridomas are cultured by a conventional method, and desired monoclonal 30 antibodies can be prepared from the culture supernatants. Alternatively, the hybridomas are administered to and grown in compatible mammals, and monoclonal antibodies are prepared from the ascites. The former method is suitable for preparing antibodies with high purity. Antibodies encoded by antibody genes that are cloned from antibody-producing cells such as the above hybridomas can also be preferably used. A cloned antibody gene is inserted 35 into an appropriate vector, and this is introduced into a host to express the antibody encoded by the gene. Methods for isolating antibody genes, inserting the genes into vectors, and 2023229507  12 Sep 2023 transforming host cells have already been established, for example, by Vandamme et al. (Eur. J. Biochem. (1990) 192(3), 767-775). Methods for producing recombinant antibodies are also known as described below. For example, a cDNA encoding the variable region (V region) of an anti-IL-6R antibody 5 is prepared from hybridoma cells expressing the anti-IL-6R antibody. For this purpose, total RNA is first extracted from hybridomas. Methods used for extracting mRNAs from cells include, for example: - the guanidine ultracentrifugation method (Biochemistry (1979) 18(24), 5294-5299), and - the AGPC method (Anal. Biochem. (1987) 162(1), 156-159) 0 Extracted mRNAs can be purified using the mRNA Purification Kit (GE Healthcare Bioscience) or such. Alternatively, kits for extracting total mRNA directly from cells, such as the QuickPrep mRNA Purification Kit (GE Healthcare Bioscience), are also commercially available. mRNAs can be prepared from hybridomas using such kits. cDNAs encoding the antibody V region can be synthesized from the prepared mRNAs using a reverse transcriptase. 5 cDNAs can be synthesized using the AMV Reverse Transcriptase First-strand cDNA Synthesis Kit (Seikagaku Co.) or such. Furthermore, the SMART RACE cDNA amplification kit (Clontech) and the PCR-based 5’-RACE method (Proc. Natl. Acad. Sci. U.S.A. (1988) 85(23), 8998-9002; Nucleic Acids Res. (1989) 17(8), 2919-2932) can be appropriately used to synthesize and amplify cDNAs. In such a cDNA synthesis process, appropriate restriction enzyme sites 0 described below may be introduced into both ends of a cDNA. The cDNA fragment of interest is purified from the resulting PCR product, and then this is ligated to a vector DNA. A recombinant vector is thus constructed, and introduced into E. coli or such. After colony selection, the desired recombinant vector can be prepared from the colony-forming E. coli. Then, whether the recombinant vector has the cDNA nucleotide 25 sequence of interest is tested by a known method such as the dideoxy nucleotide chain termination method. The 5’-RACE method which uses primers to amplify the variable region gene is conveniently used for isolating the gene encoding the variable region. First, a 5’-RACE cDNA library is constructed by cDNA synthesis using RNAs extracted from hybridoma cells as a 30 template. A commercially available kit such as the SMART RACE cDNA amplification kit is appropriately used to synthesize the 5’-RACE cDNA library. The antibody gene is amplified by PCR using the prepared 5’-RACE cDNA library as a template. Primers for amplifying the mouse antibody gene can be designed based on known antibody gene sequences. The nucleotide sequences of the primers vary depending on the 35 immunoglobulin subclass. Therefore, it is preferable that the subclass is determined in advance using a commercially available kit such as the Iso Strip mouse monoclonal antibody isotyping kit 2023229507  12 Sep 2023 (Roche Diagnostics). Specifically, for example, primers that allow amplification of genes encoding y1, y2a, Y2b, and y3 heavy chains and k and X light chains are used to isolate mouse IgG-encoding genes. In general, a primer that anneals to a constant region site close to the variable region is used as a 5 3’-side primer to amplify an IgG variable region gene. Meanwhile, a primer attached to a 5’ RACE cDNA library construction kit is used as a 5’-side primer. PCR products thus amplified are used to reshape immunoglobulins composed of a combination of heavy and light chains. A desired antibody can be selected using the IL-6R-binding activity of a reshaped immunoglobulin as an indicator. For example, when the 0 objective is to isolate an antibody against IL-6R, it is more preferred that the binding of the antibody to IL-6R is specific. An IL-6R-binding antibody can be screened, for example, by the following steps: (1) contacting an IL-6R-expressing cell with an antibody comprising the V region encoded by a cDNA isolated from a hybridoma; 5 (2) detecting the binding of the antibody to the IL-6R-expressing cell; and (3) selecting an antibody that binds to the IL-6R-expressing cell. Methods for detecting the binding of an antibody to IL-6R-expressing cells are known. Specifically, the binding of an antibody to IL-6R-expressing cells can be detected by the above-described techniques such as FACS. Immobilized samples of IL-6R-expressing cells are 0 appropriately used to assess the binding activity of an antibody. Preferred antibody screening methods that use the binding activity as an indicator also include panning methods using phage vectors. Screening methods using phage vectors are advantageous when the antibody genes are isolated from heavy-chain and light-chain subclass libraries from a polyclonal antibody-expressing cell population. Genes encoding the 25 heavy-chain and light-chain variable regions can be linked by an appropriate linker sequence to form a single-chain Fv (scFv). Phages presenting scFv on their surface can be produced by inserting a gene encoding scFv into a phage vector. The phages are contacted with an antigen of interest. Then, a DNA encoding scFv having the binding activity of interest can be isolated by collecting phages bound to the antigen. This process can be repeated as necessary to enrich 30 scFv having the binding activity of interest. After isolation of the cDNA encoding the V region of the anti-IL-6R antibody of interest, the cDNA is digested with restriction enzymes that recognize the restriction sites introduced into both ends of the cDNA. Preferred restriction enzymes recognize and cleave a nucleotide sequence that occurs in the nucleotide sequence of the antibody gene at a low frequency. 35 Furthermore, a restriction site for an enzyme that produces a sticky end is preferably introduced into a vector to insert a single-copy digested fragment in the correct orientation. The cDNA 2023229507  12 Sep 2023 encoding the V region of the anti-IL-6R antibody is digested as described above, and this is inserted into an appropriate expression vector to construct an antibody expression vector. In this case, if a gene encoding the antibody constant region (C region) and a gene encoding the above V region are fused in-frame, a chimeric antibody is obtained. Herein, “chimeric antibody” 5 means that the origin of the constant region is different from that of the variable region. Thus, in addition to mouse / human heterochimeric antibodies, human / human allochimeric antibodies are included in the chimeric antibodies of the present invention. A chimeric antibody expression vector can be constructed by inserting the above V region gene into an expression vector that already has the constant region. Specifically, for example, a recognition sequence 0 for a restriction enzyme that excises the above V region gene can be appropriately placed on the 5’ side of an expression vector carrying a DNA encoding a desired antibody constant region. A chimeric antibody expression vector is constructed by fusing in frame the two genes digested with the same combination of restriction enzymes. To produce an anti-IL-6R monoclonal antibody, antibody genes are inserted into an 5 expression vector so that the genes are expressed under the control of an expression regulatory region. The expression regulatory region for antibody expression includes, for example, enhancers and promoters. Furthermore, an appropriate signal sequence may be attached to the amino terminus so that the expressed antibody is secreted to the outside of cells. In the Examples below, a peptide having the amino acid sequence MGWSCIILFLVATATGVHS (SEQ 0 ID NO: 3) is used as a signal sequence. Meanwhile, other appropriate signal sequences may be attached. The expressed polypeptide is cleaved at the carboxyl terminus of the above sequence, and the resulting polypeptide is secreted to the outside of cells as a mature polypeptide. Then, appropriate host cells are transformed with the expression vector, and recombinant cells expressing the anti-IL-6R antibody-encoding DNA are obtained. 25 DNAs encoding the antibody heavy chain (H chain) and light chain (L chain) are separately inserted into different expression vectors to express the antibody gene. An antibody molecule having the H and L chains can be expressed by co-transfecting the same host cell with vectors into which the H-chain and L-chain genes are respectively inserted. Alternatively, host cells can be transformed with a single expression vector into which DNAs encoding the H and L 30 chains are inserted (see WO 1994 / 011523). There are various known host cell / expression vector combinations for antibody preparation by introducing isolated antibody genes into appropriate hosts. All of these expression systems are applicable to isolation of the antigen-binding domains of the present invention. Appropriate eukaryotic cells used as host cells include animal cells, plant cells, and 35 fungal cells. Specifically, the animal cells include, for example, the following cells. (1) mammalian cells: CHO (Chinese hamster ovary cell line), COS (Monkey kidney cell line), 2023229507  12 Sep 2023 myeloma (Sp2 / 0, NS0, etc.), BHK (baby hamster kidney cell line), HeLa, Vero, HEK293 (human embryonic kidney cell line with sheared adenovirus (Ad)5 DNA), PER.C6 cell (human embryonic retinal cell line transformed with the Adenovirus Type 5 (Ad5) E1A and E1B genes) and such (Current Protocols in Protein Science (May, 2001, Unit 5.9, Table 5.9.1)); 5 (2) amphibian cells: Xenopus oocytes, or such; and (3) insect cells: sf9, sf21, Tn5, or such. In addition, as a plant cell, an antibody gene expression system using cells derived from the Nicotiana genus such as Nicotiana tabacum is known. Callus cultured cells can be appropriately used to transform plant cells. 0 Furthermore, the following cells can be used as fungal cells: - yeasts: the Saccharomyces genus such as Saccharomyces serevisiae, and the Pichia genus such as Pichia pastoris; and - filamentous fungi: the Aspergillus genus such as Aspergillus niger. Furthermore, antibody gene expression systems that utilize prokaryotic cells are also 5 known. For example, when using bacterial cells, E. coli cells, Bacillus subtilis cells, and such can suitably be utilized in the present invention. Expression vectors carrying the antibody genes of interest are introduced into these cells by transfection. The transfected cells are cultured in vitro, and the desired antibody can be prepared from the culture of transformed cells. In addition to the above-described host cells, transgenic animals can also be used to 0 produce a recombinant antibody. That is, the antibody can be obtained from an animal into which the gene encoding the antibody of interest is introduced. For example, the antibody gene can be constructed as a fusion gene by inserting in frame into a gene that encodes a protein produced specifically in milk. Goat p-casein or such can be used, for example, as the protein secreted in milk. DNA fragments containing the fused gene inserted with the antibody gene is 25 injected into a goat embryo, and then this embryo is introduced into a female goat. Desired antibodies can be obtained as a protein fused with the milk protein from milk produced by the transgenic goat born from the embryo-recipient goat (or progeny thereof). In addition, to increase the volume of milk containing the desired antibody produced by the transgenic goat, hormones can be administered to the transgenic goat as necessary (Ebert, K. M. et al., 30 Bio / Technology (1994) 12 (7), 699-702). When an antigen-binding molecule described herein is administered to human, an antigen-binding domain derived from a genetically recombinant antibody that has been artificially altered to reduce the heterologous antigenicity against human and such, can be appropriately used as the antigen-binding domain of the antigen-binding molecule. Such 35 genetically recombinant antibodies include, for example, humanized antibodies. These altered antibodies are appropriately produced by known methods. 2023229507  12 Sep 2023 An antibody variable region used to produce the antigen-binding domain of an antigen-binding molecule described herein is generally formed by three complementarity-determining regions (CDRs) that are separated by four framework regions (FRs). CDR is a region that substantially determines the binding specificity of an antibody. 5 The amino acid sequences of CDRs are highly diverse. On the other hand, the FR-forming amino acid sequences often have high identity even among antibodies with different binding specificities. Therefore, generally, the binding specificity of a certain antibody can be introduced to another antibody by CDR grafting. A humanized antibody is also called a reshaped human antibody. Specifically, 0 humanized antibodies prepared by grafting the CDR of a non-human animal antibody such as a mouse antibody to a human antibody and such are known. Common genetic engineering techniques for obtaining humanized antibodies are also known. Specifically, for example, overlap extension PCR is known as a method for grafting a mouse antibody CDR to a human FR. In overlap extension PCR, a nucleotide sequence encoding a mouse antibody CDR to be grafted 5 is added to primers for synthesizing a human antibody FR. Primers are prepared for each of the four FRs. It is generally considered that when grafting a mouse CDR to a human FR, selecting a human FR that has high identity to a mouse FR is advantageous for maintaining the CDR function. That is, it is generally preferable to use a human FR comprising an amino acid sequence which has high identity to the amino acid sequence of the FR adjacent to the mouse 0 CDR to be grafted. Nucleotide sequences to be ligated are designed so that they will be connected to each other in frame. Human FRs are individually synthesized using the respective primers. As a result, products in which the mouse CDR-encoding DNA is attached to the individual FR-encoding DNAs are obtained. Nucleotide sequences encoding the mouse CDR of each 25 product are designed so that they overlap with each other. Then, complementary strand synthesis reaction is conducted to anneal the overlapping CDR regions of the products synthesized using a human antibody gene as template. Human FRs are ligated via the mouse CDR sequences by this reaction. The full length V region gene, in which three CDRs and four FRs are ultimately ligated, 30 is amplified using primers that anneal to its 5’- or 3’-end, which are added with suitable restriction enzyme recognition sequences. An expression vector for humanized antibody can be produced by inserting the DNA obtained as described above and a DNA that encodes a human antibody C region into an expression vector so that they will ligate in frame. After the recombinant vector is transfected into a host to establish recombinant cells, the recombinant cells 35 are cultured, and the DNA encoding the humanized antibody is expressed to produce the humanized antibody in the cell culture (see, European Patent Publication No. EP 239400 and 2023229507  12 Sep 2023 International Patent Publication No. WO 1996 / 002576). By qualitatively or quantitatively measuring and evaluating the antigen-binding activity of the humanized antibody produced as described above, one can suitably select human antibody FRs that allow CDRs to form a favorable antigen-binding site when ligated through the CDRs. 5 Amino acid residues in FRs may be substituted as necessary, so that the CDRs of a reshaped human antibody form an appropriate antigen-binding site. For example, amino acid sequence mutations can be introduced into FRs by applying the PCR method used for grafting a mouse CDR into a human FR. More specifically, partial nucleotide sequence mutations can be introduced into primers that anneal to the FR. Nucleotide sequence mutations are introduced 0 into the FRs synthesized by using such primers. Mutant FR sequences having the desired characteristics can be selected by measuring and evaluating the activity of the amino acid-substituted mutant antibody to bind to the antigen by the above-mentioned method (Cancer Res. (1993) 53: 851-856). Alternatively, desired human antibodies can be obtained by immunizing transgenic 5 animals having the entire repertoire of human antibody genes (see WO 1993 / 012227; WO 1992 / 003918; WO 1994 / 002602; WO 1994 / 025585; WO 1996 / 034096; WO 1996 / 033735) by DNA immunization. Furthermore, techniques for preparing human antibodies by panning using human antibody libraries are also known. For example, the V region of a human antibody is expressed 0 as a single-chain antibody (scFv) on phage surface by the phage display method. Phages expressing an scFv that binds to the antigen can be selected. The DNA sequence encoding the human antibody V region that binds to the antigen can be determined by analyzing the genes of selected phages. The DNA sequence of the scFv that binds to the antigen is determined. An expression vector is prepared by fusing the V region sequence in frame with the C region 25 sequence of a desired human antibody, and inserting this into an appropriate expression vector. The expression vector is introduced into cells appropriate for expression such as those described above. The human antibody can be produced by expressing the human antibody-encoding gene in the cells. These methods are already known (see WO 1992 / 001047; WO 1992 / 020791; WO 1993 / 006213; WO 1993 / 011236; WO 1993 / 019172; WO 1995 / 001438; WO 1995 / 015388). 30 In addition to the techniques described above, techniques of B cell cloning (identification of each antibody-encoding sequence, cloning and its isolation; use in constructing expression vector in order to prepare each antibody (IgG1, IgG2, IgG3, or IgG4 in particular); and such) such as described in Bernasconi et al. (Science (2002) 298: 2199-2202) or in WO 2008 / 081008 can be appropriately used to isolate antibody genes. 35 EU numbering and Kabat numbering 2023229507  12 Sep 2023 According to the methods used in the present invention, amino acid positions assigned to antibody CDR and FR are specified according to Kabat’s numbering (Sequences of Proteins of Immunological Interest (National Institute of Health, Bethesda, Md., 1987 and 1991)). Herein, when an antigen-binding molecule is an antibody or antigen-binding fragment, variable region 5 amino acids are indicated by Kabat numbering, while constant region amino acids are indicated by EU numbering based on Kabat’s amino acid positions. Antigen-binding domains dependent on a target tissue-specific compound To obtain an antigen-binding domain (or an antigen-binding molecule containing the 0 domain) whose antigen-binding activity varies depending on the concentration of a target tissue-specific compound, or more specifically, an antigen-binding domain (or an antigen-binding molecule containing the domain) dependent on a target tissue-specific compound, the methods indicated in the above section on binding activity may be appropriately applied. As a non-limiting embodiment, some specific examples of the methods are presented 5 below. For example, to confirm that the antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule containing the domain) in the presence of a target tissue-specific compound becomes higher than the antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule containing the domain) in the absence of the compound, the antigen-binding activities of the antigen-binding domain (or the antigen-binding molecule 0 containing the domain) in the presence and absence of the target tissue-specific compound or in the presence of high and low concentrations of the compound are compared. In another non-limiting embodiment, for example, to confirm that the antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule containing the domain) in the presence of a high concentration of a target tissue-specific compound becomes higher than the 25 antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule containing the domain) in the presence of a low concentration of the compound, the antigen-binding activities of the antigen-binding domain (or the antigen-binding molecule containing the domain) in the presence of high and low concentrations of the target tissue-specific compound are compared. 30 Furthermore, in the present invention, the phrase "the antigen-binding activity in the presence of a target tissue-specific compound is higher than the antigen-binding activity in the absence of the compound" can be alternatively expressed as "the antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule containing the domain) in the absence of a target tissue-specific compound is lower than the antigen-binding activity in the presence of 35 the compound". Furthermore, in the present invention, "the antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule containing the domain) in the absence 2023229507  12 Sep 2023 of a target tissue-specific compound is lower than the antigen-binding activity in the presence of the compound" may be alternatively described as "the antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule containing the domain) in the absence of a target tissue-specific compound is weaker than the antigen-binding activity in the presence 5 of the compound". Furthermore, in the present invention, the phrase "the antigen-binding activity in the presence of a high concentration of a target tissue-specific compound is higher than the antigen-binding activity in the presence of a low concentration of the compound" can be alternatively expressed as "the antigen-binding activity of an antigen-binding domain (or an 0 antigen-binding molecule containing the domain) in the presence of a low concentration of a target tissue-specific compound is lower than the antigen-binding activity in the presence of a high concentration of the compound". In the present invention, "the antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule containing the domain) in the presence of a low concentration of a target tissue-specific compound is lower than the 5 antigen-binding activity in the presence of a high concentration of the compound" may be alternatively described as "the antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule containing the domain) in the presence of a low concentration of a target tissue-specific compound is weaker than the antigen-binding activity in the presence of a high concentration of the compound". 0 Conditions when measuring antigen-binding activity other than the concentration of a target tissue-specific compound are not particularly limited, and can be selected appropriately by those skilled in the art. For example, it is possible to measure under conditions of HEPES buffer and 37°C. For example, Biacore (GE Healthcare) or such can be used for measurement. When the antigen is a soluble molecule, the activity of an antigen-binding domain (or an 25 antigen-binding molecule containing the domain) to bind to the soluble molecule can be determined by loading the antigen as an analyte onto a chip immobilized with the antigen-binding domain (or an antigen-binding molecule containing the domain). Alternatively, when the antigen is a membrane-type molecule, the binding activity towards the membrane-type molecule can be determined by loading the antigen-binding domain (or an antigen-binding 30 molecule containing the domain) as an analyte onto a chip immobilized with the antigen. As long as the antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule containing the domain) contained in antigen-binding molecules of the present invention in the absence of a target tissue-specific compound is weaker than the antigen-binding activity in the presence of the target tissue-specific compound, the ratio between 35 the antigen-binding activity in the absence of the compound and the antigen-binding activity in the presence of the compound is not particularly limited. However, the value of KD (in the 2023229507  12 Sep 2023 absence of the compound) / KD (in the presence of the compound), which is a ratio of dissociation constant (KD) against an antigen in the absence of the target tissue-specific compound to KD in the presence of the compound, is preferably 2 or greater, more preferably 10 or greater, and still more preferably 40 or greater. The upper limit of the value of KD (in the 5 absence of the compound) / KD (in the presence of the compound) is not particularly limited, and may be any value, for example, 400, 1,000, or 10,000, as long as it can be provided by the technologies of those skilled in the art. When antigen-binding activity is not observed in the absence of the target tissue-specific compound, the value of the upper limit is infinity. As long as the antigen-binding activity of an antigen-binding domain (or an 0 antigen-binding molecule containing the domain) contained in antigen-binding molecules of the present invention in the presence of a low concentration of a target tissue-specific is weaker than the antigen-binding activity in the presence of a high concentration of the target tissue-specific compound, the ratio between the antigen-binding activity in the presence of a low concentration of the compound and the antigen-binding activity in the presence of a high concentration of the 5 compound is not particularly limited. However, the value of KD (in the presence of a low concentration of the compound) / KD (in the presence of a high concentration of the compound), which is a ratio of dissociation constant (KD) against an antigen in the presence of a low concentration of the target tissue-specific compound to KD in the presence of a high concentration of the compound, is preferably 2 or greater, more preferably 10 or greater, and still 0 more preferably 40 or greater. The upper limit of the value of KD (in the presence of a low concentration of the compound) / KD (in the presence of a high concentration of the compound) is not particularly limited, and may be any value, for example, 400, 1,000, or 10,000, as long as it can be provided by the technologies of those skilled in the art. When antigen-binding activity is not observed in the presence of a low concentration of the target tissue-specific compound, the 25 value of the upper limit is infinity. For the value of antigen-binding activity, if the antigen is a soluble molecule, dissociation constant (KD) can be used; and if the antigen is a membrane-type molecule, apparent dissociation constant (apparent KD) can be used. The dissociation constant (KD) and apparent dissociation constant (apparent KD) can be determined by methods known to those 30 skilled in the art, for example, using Biacore (GE Healthcare), a Scatchard plot, a flow cytometer, or such. As another indicator that shows the ratio between the antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule containing the domain) of the present invention in the absence of a target tissue-specific compound and the antigen-binding activity in 35 the presence of the compound, for example, dissociation rate constant kd can be suitably used. When the dissociation rate constant (kd) is used instead of the dissociation constant (KD) as an 2023229507  12 Sep 2023 indicator that shows the binding activity ratio, the value of kd (in the absence of the compound) / kd (in the presence of the compound), which is a ratio between kd (dissociation rate constant) for an antigen in the absence of a target tissue-specific compound and kd in the presence of the compound, is preferably 2 or greater, more preferably 5 or greater, even more preferably 10 or 5 greater, and still more preferably 30 or greater. The upper limit of the value of kd (in the absence of the compound) / kd (in the presence of the compound) is not particularly limited, and may be any value, for example, 50, 100, or 200, as long as it can be provided by the common technical knowledge of those skilled in the art. When antigen-binding activity is not observed in the absence of the tissue-specific compound, there is no dissociation and the value of the 0 upper limit becomes infinity. As another indicator that shows the ratio between the antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule containing the domain) of the present invention in the presence of a low concentration of a target tissue-specific compound and the antigen-binding activity in the presence of a high concentration of the compound, for example, 5 dissociation rate constant kd can be suitably used. When the dissociation rate constant (kd) is used instead of the dissociation constant (KD) as an indicator showing the binding activity ratio, the value of kd (in the presence of a low concentration of the compound) / kd (in the presence of a high concentration of the compound), which is a ratio between kd (dissociation rate constant) for an antigen in the presence of a low concentration of a target tissue-specific compound and kd 0 in the presence of a high concentration of the compound, is preferably 2 or greater, more preferably 5 or greater, even more preferably 10 or greater, and still more preferably 30 or greater. The upper limit of the value of kd (in the presence of a low concentration of the compound) / kd (in the presence of a high concentration of the compound) is not particularly limited, and may be any value, for example, 50, 100, or 200, as long as it can be provided by the common technical 25 knowledge of those skilled in the art. When antigen-binding activity is not observed in the presence of a low concentration of the target tissue-specific compound, there is no dissociation and the value of the upper limit becomes infinity. For the value of antigen-binding activity, if the antigen is a soluble molecule, dissociation rate constant (kd) can be used; and if the antigen is a membrane-type molecule, 30 apparent dissociation rate constant (apparent kd) can be used. The dissociation rate constant (kd) and apparent dissociation rate constant (apparent kd) can be determined by methods known to those skilled in the art, for example, using Biacore (GE Healthcare), a flow cytometer, or such. In the present invention, when measuring the antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule containing the domain) at a certain concentration of the 35 target tissue-specific compound, conditions other than the concentration of the compound concentration are preferably the same. 2023229507  12 Sep 2023 For example, in an embodiment provided by the present invention, an antigen-binding domain (or an antigen-binding molecule containing the domain) with lower antigen-binding activity in the absence of a target tissue-specific compound than in the presence of the compound, may be obtained by screening of antigen-binding domains (or antigen-binding molecules) that 5 comprises the steps of: (a) determining antigen-binding activity of antigen-binding domains (or antigen-binding molecules) in the absence of a target tissue-specific compound; (b) determining antigen-binding activity of the antigen-binding domains (or antigen-binding molecules) in the presence of the target tissue-specific compound; and 0 (c) selecting an antigen-binding domain (or an antigen-binding molecule) with lower antigen-binding activity in the absence of the target tissue-specific compound than in the presence of the compound. For example, in an embodiment provided by the present invention, an antigen-binding domain (or an antigen-binding molecule containing the domain) with lower antigen-binding 5 activity in the presence of a low concentration of a target tissue-specific compound than in the presence of a high concentration of the compound, may be obtained by screening of antigen-binding domains (or antigen-binding molecules) that comprises the steps of: (a) determining antigen-binding activity of antigen-binding domains (or antigen-binding molecules) in the presence of a low concentration of a target tissue-specific compound; 0 (b) determining antigen-binding activity of the antigen-binding domains (or antigen-binding molecules) in the presence of a high concentration of the target tissue-specific compound; and (c) selecting an antigen-binding domain (or an antigen-binding molecule) with lower antigen-binding activity in the presence of a low concentration of the target tissue-specific compound than in the presence of a high concentration of the compound. 25 Furthermore, in an embodiment provided by the present invention, an antigen-binding domain (or an antigen-binding molecule containing the domain) with lower antigen-binding activity in the absence of a target tissue-specific compound than in the presence of the compound, may be obtained by screening of antigen-binding domains (or antigen-binding molecules) or a library thereof that comprises the steps of: 30 (a) contacting antigen-binding domains (or antigen-binding molecules) or a library thereof with an antigen in the presence of a target tissue-specific compound; (b) placing antigen-binding domains (or antigen-binding molecules) that bind to the antigen in said step (a) in the absence of the compound; (c) isolating an antigen-binding domain (or an antigen-binding molecule) that dissociated in 35 said step (b). Furthermore, in an embodiment provided by the present invention, an antigen-binding 2023229507  12 Sep 2023 domain (or an antigen-binding molecule containing the domain) with lower antigen-binding activity in the presence of a low concentration of a target tissue-specific compound than in the presence of a high concentration of the compound, may be obtained by screening of antigen-binding domains (or antigen-binding molecules) or a library thereof that comprises the 5 steps of: (a) contacting antigen-binding domains (or antigen-binding molecules) or a library thereof with an antigen in the presence of a high concentration of a target tissue-specific compound; (b) placing antigen-binding domains (or antigen-binding molecules) that bind to the antigen in said step (a) in the presence of a low concentration of the compound; 0 (c) isolating an antigen-binding domain (or an antigen-binding molecule) that dissociates in said step (b). Alternatively, in an embodiment provided by the present invention, an antigen-binding domain (or an antigen-binding molecule containing the domain) with lower antigen-binding activity in the absence of a target tissue-specific compound than in the presence of the compound, 5 may be obtained by screening of antigen-binding domains (or antigen-binding molecules) or a library thereof that comprises the steps of: (a) contacting a library of antigen-binding domains (or antigen-binding molecules) with an antigen in the absence of a target tissue-specific compound; (b) selecting antigen-binding domains (or antigen-binding molecules) that do not bind to the 0 antigen in said step (a); (c) allowing the antigen-binding domains (or antigen-binding molecules) selected in said step (b) to bind to the antigen in the presence of the compound; and (d) isolating an antigen-binding domain (or an antigen-binding molecule) that binds to the antigen in said step (c). 25 Alternatively, in an embodiment provided by the present invention, an antigen-binding domain (or an antigen-binding molecule containing the domain) with lower antigen-binding activity in the presence of a low concentration of a target tissue-specific compound than in the presence of a high concentration of the compound, may be obtained by screening of antigen-binding domains (or antigen-binding molecules) or a library thereof that comprises the 30 steps of: (a) contacting a library of antigen-binding domains (or antigen-binding molecules) with an antigen in the presence of a low concentration of a target tissue-specific compound; (b) selecting antigen-binding domains (or antigen-binding molecules) that do not bind to the antigen in said step (a); 35 (c) allowing the antigen-binding domains (or antigen-binding molecules) selected in said step (b) to bind to the antigen in the presence of a high concentration the compound; and 2023229507  12 Sep 2023 (d) isolating an antigen-binding domain (or an antigen-binding molecule) that binds to the antigen in said step (c). Furthermore, in an embodiment provided by the present invention, an antigen-binding domain (or an antigen-binding molecule containing the domain) with lower antigen-binding 5 activity in the absence of a target tissue-specific compound than in the presence of the compound, may be obtained by a screening method comprising the steps of: (a) contacting a library of antigen-binding domains (or antigen-binding molecules) with an antigen-immobilized column in the presence of a target tissue-specific compound; (b) eluting an antigen-binding domain (or antigen-binding molecule) that binds to the column in 0 said step (a) from the column in the absence of the compound; and (c) isolating the antigen-binding domain (or antigen-binding molecule) eluted in said step (b). Furthermore, in an embodiment provided by the present invention, an antigen-binding domain (or an antigen-binding molecule containing the domain) with lower antigen-binding activity in the presence of a low concentration of a target tissue-specific compound than in the 5 presence of a high concentration of the compound, may be obtained by a screening method comprising the steps of: (a) contacting a library of antigen-binding domains (or antigen-binding molecules) with an antigen-immobilized column in the presence of a high concentration of a target tissue-specific compound; 0 (b) eluting an antigen-binding domain (or antigen-binding molecule) that binds to the column in said step (a) from the column in the presence of a low concentration of the compound; and (c) isolating the antigen-binding domain (or antigen-binding molecule) eluted in said step (b). Furthermore, in an embodiment provided by the present invention, an antigen-binding domain (or an antigen-binding molecule containing the domain) with lower antigen-binding 25 activity in the absence of a target tissue-specific compound than in the presence of the compound, may be obtained by a screening method comprising the steps of: (a) allowing a library of antigen-binding domains (or antigen-binding molecules) to pass through an antigen-immobilized column in the absence of a target tissue-specific compound; (b) collecting an antigen-binding domain (or antigen-binding molecule) eluted without binding 30 to the column in said step (a); (c) allowing the antigen-binding domain (or antigen-binding molecule) collected in said step (b) to bind to the antigen in the presence of the compound; and (d) isolating an antigen-binding domain (or antigen-binding molecule) that binds to the antigen in said step (c). 35 Furthermore, in an embodiment provided by the present invention, an antigen-binding domain (or an antigen-binding molecule containing the domain) with lower antigen-binding 2023229507  12 Sep 2023 activity in the presence of a low concentration of a target tissue-specific compound than in the presence of a high concentration of the compound, may be obtained by a screening method comprising the steps of: (a) allowing a library of antigen-binding domains (or antigen-binding molecules) to pass 5 through an antigen-immobilized column in the presence of a low concentration of a target tissue-specific compound; (b) collecting an antigen-binding domain (or antigen-binding molecule) eluted without binding to the column in said step (a); (c) allowing the antigen-binding domain (or antigen-binding molecule) collected in said step (b) 0 to bind to the antigen in the presence of a high concentration of the compound; and (d) isolating an antigen-binding domain (or antigen-binding molecule) that binds to the antigen in said step (c). Furthermore, in an embodiment provided by the present invention, an antigen-binding domain (or an antigen-binding molecule containing the domain) with lower antigen-binding 5 activity in the absence of a target tissue-specific compound than in the presence of the compound, may be obtained by a screening method comprising the steps of: (a) contacting an antigen with a library of antigen-binding domains (or antigen-binding molecules) in the presence of a target tissue-specific compound; (b) obtaining an antigen-binding domain (or antigen-binding molecule) that binds to the antigen 0 in said step (a); (c) placing the antigen-binding domain (or antigen-binding molecule) obtained in said step (b) in the absence of the compound; and (d) isolating an antigen-binding domain (or antigen-binding molecule) whose antigen-binding activity in said step (c) is weaker than that of the reference selected in said step (b). 25 Furthermore, in an embodiment provided by the present invention, an antigen-binding domain (or an antigen-binding molecule containing the domain) with lower antigen-binding activity in the presence of a low concentration of a target tissue-specific compound than in the presence of a high concentration of the compound, may be obtained by a screening method comprising the steps of: 30 (a) contacting an antigen with a library of antigen-binding domains (or antigen-binding molecules) in the presence of a high concentration of a target tissue-specific compound; (b) obtaining an antigen-binding domain (or antigen-binding molecule) that binds to the antigen in said step (a); (c) placing the antigen-binding domain (or antigen-binding molecule) obtained in said step (b) in 35 the presence of a low concentration of the compound; and (d) isolating an antigen-binding domain (or antigen-binding molecule) whose antigen-binding 2023229507  12 Sep 2023 activity in said step (c) is weaker than that of the reference selected in said step (b). The above-mentioned steps may be repeated two or more times. Thus, the present invention provides an antigen-binding domain (or an antigen-binding molecule containing the domain) with lower antigen-binding activity in the absence of a target tissue-specific compound 5 than in the presence of the compound, or an antigen-binding domain (or an antigen-binding molecule containing the domain) with lower antigen-binding activity in the presence of a low concentration of a target tissue-specific compound than in the presence of a high concentration of the compound, obtained by screening methods that further comprise the step of repeating steps (a) to (c) or (a) to (d) two or more times in the above-mentioned screening methods. The 0 number of repeats of steps (a) to (c) or (a) to (d) is not particularly limited, and it is generally ten or less. In the screening methods of the present invention, a target tissue-specific compound may be a compound defined by quantitative target tissue specificity such as presence in the target tissue at a concentration (for example, high concentration or low concentration) different from 5 the concentration in non-target tissues. For example, a target tissue-specific compound is differentially present at any concentrations. However, generally, a target tissue-specific compound can be present at a concentration increased by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at 0 least 90%, at least 95%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 2-fold, at least 5-fold, at least 10-fold, at least 50-fold, at least 100-fold, at least 103-fold, at least 104-fold, at least 105-fold, at least 106-fold, or more, or up to infinity (when the compound is absent in non-target tissues). The threshold differentiating low and high concentrations can be set appropriately 25 according to the compound. For example, in a non-limiting embodiment of the threshold of ATP or adenosine, the threshold for a low-concentration condition may be selected appropriately from the values of 10 nM, 1 nM, 100 pM, 10 pM, 1 pM, and 0 M. Depending on the predetermined threshold, the high-concentration condition may be set appropriately at a value selected from at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 30 twice, at least five-fold, at least 10-fold, at least 50-fold, at least 100-fold, at least 103-fold, at least 104-fold, at least 105-fold, and at least 106-fold the value of each threshold. Furthermore, in a non-limiting embodiment of PGE2, the threshold for a low-concentration condition may be selected appropriately from the values of 10 pM, 1 pM, 100 fM, 10 fM, 1 fM, and 0 M. Depending on the predetermined threshold, the high-concentration condition may be set 35 appropriately at a value selected from at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least twofold, at least five-fold, at least 10-fold, at least 50-fold, at least 2023229507  12 Sep 2023 100-fold, at least 103-fold, at least 104-fold, at least 105-fold, and at least 106-fold the value of each threshold. Furthermore, in a non-limiting embodiment of Kynurenine, the threshold for a low-concentration condition may be selected appropriately from the values of10 pM, 1 pM, 100 nM, 10 nM, and 1 nM, and 0 M. Depending on the predetermined threshold, the 5 high-concentration condition may be set appropriately at a value selected from at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least twofold, at least five-fold, at least 10-fold, at least 50-fold, at least 100-fold, at least 103-fold, at least 104-fold, at least 105-fold, and at least 106-fold the value of each threshold. The antigen-binding activity of an antigen-binding domain (or an antigen-binding 0 molecule) may be measured by a method known to those skilled in the art, and conditions other than the concentration of a target tissue-specific compound can be set appropriately by one skilled in the art. The antigen-binding activity of an antigen-binding domain (or an antigen-binding molecule) can be assessed as dissociation constant (KD), apparent dissociation constant (apparent KD), dissociation rate constant (kd), apparent dissociation rate constant 5 (apparent kd), etc. They can be determined by methods known to those skilled in the art, for example, using Biacore (GE Healthcare), the Scatchard plot, FACS, or such. In the present invention, the step of selecting an antibody or an antigen-binding domain with higher antigen-binding activity in the presence of a target tissue-specific compound than in the absence of the compound has the same meaning as the step of selecting an antibody or an 0 antigen-binding domain with lower antigen-binding activity in the absence of a target tissue-specific compound than in the presence of the compound. In the present invention, the step of selecting an antibody or an antigen-binding domain with higher antigen-binding activity in the presence of a high concentration of a target tissue-specific compound than in the presence of a low concentration of the compound has the 25 same meaning as the step of selecting an antibody or an antigen-binding domain with lower antigen-binding activity in the absence of a target tissue-specific compound than in the presence of the compound. As long as antigen-binding activity in the absence of a target tissue-specific compound is lower than the antigen-binding activity in the presence of the compound, the difference 30 between antigen-binding activity in the presence of the compound and antigen-binding activity in the absence of the compound is not particularly limited, but preferably, the antigen-binding activity in the presence of the compound relative to the antigen-binding activity in the absence of the compound is twofold or more, more preferably 10-fold or more, and even more preferably 40-fold or more. The upper limit of the difference between the antigen-binding activities is not 35 particularly limited, and as long as it can be produced by the techniques of those skilled in the art, any value such as 400-fold, 1000-fold, or 10000-fold is possible. In the absence of a target 2023229507  12 Sep 2023 tissue-specific compound, when antigen-binding activity is not observed, this upper limit becomes infinity. The antigen-binding domains (or antigen-binding molecules containing the domains) of the present invention which are to be screened by the aforementioned screening methods may be 5 any antigen-binding domains (or antigen-binding molecules); and for example, the above-mentioned antigen-binding domains (or antigen-binding molecules) can be screened. For example, antigen-binding domains (or antigen-binding molecules) having naturally-occurring sequences can be screened, and antigen-binding domains (or antigen-binding molecules) with substituted amino acid sequences may be screened. 0 Library According to a certain embodiment, the antigen-binding domain (or an antigen-binding molecule containing this domain) of the present invention can be obtained from a library mainly comprising a plurality of antigen-binding molecules having different sequences from one another, 5 in which at least one amino acid residue that changes the binding activity of the antigen-binding molecule toward an antigen dependent on a target tissue-specific compound is contained in the antigen-binding domain. Examples of the compound include (1) primary metabolites of the Krebs cycle or the glycolytic pathway such as lactose, succinic acid, or citric acid, (2) amino acids such as alanine, glutamic acid, or asparagine, (3) kynurenine and amino acid metabolites 0 thereof such as anthranilic acid, 3-hudroxykynurenine, and kynurenic acid, (4) arachidonic acid metabolites such as prostaglandin E2, and (5) nucleosides carrying a purine ring structure such as adenosine, adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine monophosphate (AMP). Below are examples of such a library mainly comprising a plurality of antigen-binding molecules having different sequences from one another, in which at least one 25 amino acid residue that changes the binding activity of the antigen-binding molecule toward adenosine- and / or ATP-dependent antigens which are target tissue-specific compounds is contained in the antigen-binding domain. Herein, a “library” refers to a plurality of antigen-binding molecules or a plurality of fusion polypeptides containing antigen-binding molecules, or nucleic acids or polynucleotides 30 encoding their sequences. The sequences of a plurality of antigen-binding molecules or a plurality of fusion polypeptides containing antigen-binding molecules in a library are not identical, but are different from one another. Herein, the phrase “sequences are different from one another” in the expression “a plurality of antigen-binding molecules whose sequences are different from one another” means 35   that the sequences of antigen-binding molecules in a library are different from one another. Specifically, in a library, the number of sequences different from one another reflects the number 2023229507  12 Sep 2023 of independent clones with different sequences, and may also be referred to as “library size”. The library size of a conventional phage display library ranges from 106 to 1012. The library size can be increased up to 1014 by the use of known techniques such as ribosome display. However, the actual number of phage particles used in panning selection of a phage library is in 5 general 10 to 10,000 times greater than the library size. This excess multiplicity is also referred to as “the number of library equivalents”, and means that there are 10 to 10,000 individual clones that have the same amino acid sequence. Thus, in the present invention, the phrase “sequences are different from one another” means that the sequences of independent antigen-binding molecules in a library, excluding library equivalents, are different from one another. More 0 specifically, the above means that there are 106 to 1014 antigen-binding molecules whose sequences are different from one another, preferably 107 to 1012 molecules, more preferably 108 to 1011 molecules, and particularly preferably 108 to 1010 molecules whose sequences are different from one another. Herein, the phrase “a plurality of” in the expression “a library mainly composed of a 5 plurality of antigen-binding molecules” generally refers to, in the case of, for example, antigen-binding molecules, fusion polypeptides, polynucleotide molecules, vectors, or viruses of the present invention, a group of two or more types of the substance. For example, when two or more substances are different from one another in a particular characteristic, this means that there are two or more types of the substance. Such examples may include, for example, mutant 0 amino acids observed at specific amino acid positions in an amino acid sequence. For example, when there are two or more antigen-binding molecules of the present invention whose sequences are substantially the same or preferably the same except for flexible residues or except for particular mutant amino acids at hypervariable positions exposed on the surface, there are a plurality of antigen-binding molecules of the present invention. In another example, when there 25 are two or more polynucleotide molecules whose sequences are substantially the same or preferably the same except for nucleotides encoding flexible residues or nucleotides encoding mutant amino acids of hypervariable positions exposed on the surface, there are a plurality of polynucleotide molecules of the present invention. In addition, herein, the phrase "mainly composed of" in the expression "a library mainly 30 composed of a plurality of antigen-binding molecules" reflects the number of antigen-binding molecules whose antigen-binding activity varies depending on the concentration of a target tissue-specific compound, among independent clones with different sequences in a library. Specifically, it is preferable that there are at least 104 antigen-binding molecules having such binding activity in a library. More preferably, antigen-binding domains of the present invention 35   can be obtained from a library containing at least 105 antigen-binding molecules having such binding activity. Still more preferably, antigen-binding domains of the present invention can be 2023229507  12 Sep 2023 obtained from a library containing at least 106 antigen-binding molecules having such binding activity. Particularly preferably, antigen-binding domains of the present invention can be obtained from a library containing at least 107 antigen-binding molecules having such binding activity. Yet more preferably, antigen-binding domains of the present invention can be obtained 5 from a library containing at least 108 antigen-binding molecules having such binding activity. Alternatively, this may also be preferably expressed as the ratio of the number of antigen-binding molecules in which antigen-binding activity of the antigen-binding domain varies depending on the presence or absence of adenosine and / or ATP with respect to the number of independent clones having different sequences in a library. Specifically, antigen-binding domains of the 0 present invention can be obtained from a library in which antigen-binding molecules having such binding activity account for 0.1% to 80%, preferably 0.5% to 60%, more preferably 1% to 40%, still more preferably 2% to 20%, and particularly preferably 4% to 10% of independent clones with different sequences in the library. In the case of fusion polypeptides, polynucleotide molecules, or vectors, similar expressions may be possible using the number of molecules or the 5 ratio to the total number of molecules. In the case of viruses, similar expressions may also be possible using the number of virions or the ratio to total number of virions. Amino acids that change the antigen-binding activity of the antigen-binding domain depending on the presence or absence of adenosine and / or ATP 0 Antigen-binding domains or antibodies of the present invention screened by the above-described screening methods may be prepared in any manner. It is possible to use preexisting antibodies, preexisting libraries (phage libraries, etc.), antibodies or libraries prepared from hybridomas obtained by immunizing animals or from B cells of immunized animals, and antibodies or libraries prepared from immune cells such as B cells of animals 25 immunized by a conjugate in which adenosine or ATP is suitably linked to an adjuvant agent such as a highly immunogenic T cell epitope peptide. A non-limiting example of the T cell epitope peptide suitably includes Tetanus toxin-derived p30 helper peptide (shown in SEQ ID NO: 4, and also referred to as Fragment C (FrC)). Examples of amino acids that change the antigen-binding activity of the antigen-binding 30 molecule depending on the presence or absence of adenosine and / or ATP as described above include amino acids that form an adenosine- and / or ATP-binding motif. The amino acid positions where the above-mentioned amino acids are contained in the antigen-binding domain are not limited to any specific position. As long as the antigen-binding activity of the antigen-binding domain is changed depending on the presence or absence of adenosine and / or 35 ATP, any position in the heavy chain variable region or light chain variable region forming the antigen-binding domain is possible. More specifically, the antigen-binding domains of the 2023229507  12 Sep 2023 present invention may be obtained from a library mainly comprising antigen-binding molecules having different sequences from one another, in which the amino acids that change the antigen-binding activity of the antigen-binding molecule depending on the presence or absence of adenosine and / or ATP are contained in the antigen-binding domain of the heavy chain. In a 5 non-limiting embodiment, antigen-binding domains of the present invention may be obtained from a library mainly comprising antigen-binding molecules having different sequences from one another, in which the amino acids that change the antigen-binding activity of the antigen-binding molecule depending on the presence or absence of adenosine and / or ATP are contained in CDR1, CDR2, and / or CDR3 of the heavy chain. In another non-limiting 0 embodiment, antigen-binding domains of the present invention may be obtained from a library mainly comprising antigen-binding molecules having different sequences from one another, in which the amino acids that change the antigen-binding activity of the antigen-binding molecule depending on the presence or absence of adenosine and / or ATP are contained in FR1, FR2, FR3 and / or FR4 of the heavy chain. 5 Furthermore, in an embodiment of the present invention, antigen-binding domains of the present invention may be obtained from a library mainly comprising antigen-binding molecules having different sequences from one another, in which the amino acids that change the antigen-binding activity of the antigen-binding molecule depending on the presence or absence of adenosine and / or ATP are contained in the antigen-binding domain of the heavy chain and / or 0 light chain. In a non-limiting embodiment, antigen-binding domains of the present invention may be obtained from a library mainly comprising antigen-binding molecules having different sequences from one another, in which the amino acids that change the antigen-binding activity of the antigen-binding molecule depending on the presence or absence of adenosine and / or ATP are contained in CDR1, CDR2, and / or CDR3 of the heavy chain and / or light chain. In another 25 non-limiting embodiment, antigen-binding domains of the present invention may be obtained from a library mainly comprising antigen-binding molecules having different sequences from one another, in which the amino acids that change the antigen-binding activity of the antigen-binding molecule depending on the presence or absence of adenosine and / or ATP are contained in FR1, FR2, FR3 and / or FR4 of the heavy chain and / or light chain. 30 In a non-limiting embodiment, examples of such amino acids include any one or more amino acids selected from amino acids at positions 52, 52a, 53, 96, 100a, and 100c contained in the heavy chain variable region. Also, in a non-limiting embodiment, examples of such amino acids include one or more amino acids selected from amino acids including Ser at position 52, Ser at position 52a, Arg at position 53, Gly at position 96, Leu at position 100a, and Trp at 35 position 100c contained in the heavy chain variable region. Any framework sequence can be used as the framework sequence of the light-chain 2023229507  12 Sep 2023 and / or heavy-chain variable regions of an antigen-binding molecule as long as the amino acids that change the antigen-binding activity of the antigen-binding molecule depending on the presence or absence of adenosine and / or ATP are contained in the antigen-binding domain of the heavy chain and / or light chain. The origin of the framework sequences is not limited, and they 5 may be obtained from human or any nonhuman organisms. Such organisms preferably include mice, rats, guinea pigs, hamsters, gerbils, cats, rabbits, dogs, goats, sheep, bovines, horses, camels and organisms selected from nonhuman primates. In a particularly preferred embodiment, the framework sequences of the light chain and / or heavy chain variable region of an antigen-binding molecule preferably have human germ-line framework sequences. Thus, in 0 an embodiment of the present invention, if the entire framework sequences are human sequences, it is thought that an antigen-binding molecule of the present invention induces little or no immunogenic response when it is administered to humans (for example, to treat diseases). In the above sense, the phrase "containing a germ line sequence" in the present invention means that a part of the framework sequences of the present invention is identical to a part of any 5 human germ line framework sequences. For example, when the heavy chain FR2 sequence of an antigen-binding molecule of the present invention is a combination of heavy chain FR2 sequences of different human germ line framework sequences, such a molecule is also an antigen-binding molecule "containing a germ line sequence" in the present invention. Even when the framework sequences of antigen-binding molecules of the present invention are 0 sequences with substitutions, they are antigen-binding molecules "containing a germ line sequence" of the present invention. Examples of such sequences with substitutions include, in particular, sequences in which amino acids of part of human germ line framework sequences have been substituted with amino acids that change the antigen-binding activity of the antigen-binding molecule depending on the presence or absence of adenosine and / or ATP. 25 Preferred examples of the frameworks include, for example, fully human framework region sequences currently known, which are included in the website of V-Base (http: / / vbase.mrc-cpe.cam.ac.uk / ) or others. Those framework region sequences can be appropriately used as a germ line sequence contained in an antigen-binding molecule of the present invention. The germ line sequences may be categorized according to their similarity 30   (Tomlinson et al. (J. Mol. Biol. (1992) 227, 776-798); Williams and Winter (Eur. J. Immunol. (1993) 23, 1456-1461); Cox et al. (Nat. Genetics (1994) 7, 162-168)). Appropriate germ line sequences can be selected from Vk, which is grouped into seven subgroups; VX, which is grouped into ten subgroups; and VH, which is grouped into seven subgroups. Fully human VH sequences preferably include, but are not limited to, for example, VH 35 sequences of: subgroup VH1 (for example, VH1-2, VH1-3, VH1-8, VH1-18, VH1-24, VH1-45, VH1-46, 2023229507  12 Sep 2023 VH1-58, and VH1-69); subgroup VH2 (for example, VH2-5, VH2-26, and VH2-70); subgroup VH3 (VH3-7, VH3-9, VH3-11, VH3-13, VH3-15, VH3-16, VH3-20, VH3-21, VH3-23, VH3-30, VH3-33, VH3-35, VH3-38, VH3-43, VH3-48, VH3-49, VH3-53, VH3-64, VH3-66, 5 VH3-72, VH3-73, and VH3-74); subgroup VH4 (VH4-4, VH4-28, VH4-31, VH4-34, VH4-39, VH4-59, and VH4-61); subgroup VH5 (VH5-51); subgroup VH6 (VH6-1); and subgroup VH7 (VH7-4 and VH7-81). 0 These are also described in known documents (Matsuda et al. (J. Exp. Med. (1998) 188, 1973-1975)) and such, and thus persons skilled in the art can appropriately design antigen-binding molecules of the present invention based on the information of these sequences. It is also preferable to use other fully human frameworks or framework sub-regions. Fully human Vk sequences preferably include, but are not limited to, for example: 5 A20, A30, L1, L4, L5, L8, L9, L11, L12, L14, L15, L18, L19, L22, L23, L24, O2, O4, O8, O12, O14, and O18 grouped into subgroup Vk1; A1, A2, A3, A5, A7, A17, A18, A19, A23, O1, and O11, grouped into subgroup Vk2; A11, A27, L2, L6, L10, L16, L20, and L25, grouped into subgroup Vk3; B3, grouped into subgroup Vk4; 0 B2 (herein also referred to as Vk5-2), grouped into subgroup Vk5; and A10, A14, and A26, grouped into subgroup Vk6 (Kawasaki et al. (Eur. J. Immunol. (2001) 31, 1017-1028); Schable and Zachau (Biol. Chem. Hoppe Seyler (1993) 374, 1001-1022); Brensing-Kuppers et al. (Gene (1997) 191, 173-181)). Fully human V! sequences preferably include, but are not limited to, for example: 25 V1-2, V1-3, V1-4, V1-5, V1-7, V1-9, V1-11, V1-13, V1-16, V1-17, V1-18, V1-19, V1-20, and V1-22, grouped into subgroup VL1; V2-1, V2-6, V2-7, V2-8, V2-11, V2-13, V2-14, V2-15, V2-17, and V2-19, grouped into subgroup VL1; V3-2, V3-3, and V3-4, grouped into subgroup VL3; 30 V4-1, V4-2, V4-3, V4-4, and V4-6, grouped into subgroup VL4; and V5-1, V5-2, V5-4, and V5-6, grouped into subgroup VL5 (Kawasaki et al. (Genome Res. (1997) 7, 250-261)). Normally, these framework sequences are different from one another at one or more amino acid residues. These framework sequences can be used in combination with “at least one 35 amino acid residue that alters the antigen-binding activity of an antigen-binding domain depending on the presence or absence of adenosine and / or ATP” of the present invention. Other 2023229507  12 Sep 2023 examples of the fully human frameworks used in combination with “at least one amino acid residue that alters the antigen-binding activity of an antigen-binding domain depending on the presence or absence of adenosine and / or ATP” of the present invention include, but are not limited to, for example, KOL, NEWM, REI, EU, TUR, TEI, LAY, and POM (for example, Kabat 5   et al. (1991) supra; Wu et al. (J. Exp. Med. (1970) 132, 211-250)). Without being bound by a particular theory, one reason for the expectation that the use of germ line sequences precludes adverse immune responses in most individuals is believed to be as follows. As a result of the process of affinity maturation during normal immune responses, somatic mutation occurs frequently in the variable regions of immunoglobulin. Such mutations 0 mostly occur around CDRs whose sequences are hypervariable, but also affect residues of framework regions. Such framework mutations do not exist on the germ line genes, and also they are less likely to be immunogenic in patients. On the other hand, the normal human population is exposed to most of the framework sequences expressed from the germ line genes. As a result of immunotolerance, these germ line frameworks are expected to have low or no 5 immunogenicity in patients. To maximize the possibility of immunotolerance, variable region-encoding genes may be selected from a group of commonly occurring functional germ line genes. Known methods such as site-directed mutagenesis (Kunkel et al. (Proc. Natl. Acad. Sci. USA (1985) 82, 488-492)) and overlap extension PCR can be appropriately employed to produce 0 the antigen-binding molecules of the present invention in which the above-described variable region sequences, heavy or light chain variable region sequences, CDR sequences, or framework sequences contain amino acids that alter the antigen-binding activity of the antigen-binding domain depending on the presence or absence of adenosine and / or ATP. For example, a library which contains a plurality of antigen-binding molecules of the 25 present invention whose sequences are different from one another can be constructed by combining heavy chain variable regions prepared as a randomized variable region sequence library with a light chain variable region selected as a CDR sequence and / or framework sequence originally containing at least one amino acid residue that alters the antigen-binding activity of the antigen-binding domain depending on the presence or absence of adenosine and / or 30 ATP. Alternatively, a heavy chain and / or light chain variable region sequence selected as a CDR sequence and / or a framework sequence originally containing at least one amino acid residue that changes the antigen-binding activity of an antigen-binding domain depending on the presence or absence of adenosine and / or ATP as mentioned above, can be designed to contain 35 various amino acid residues other than the above amino acid residue(s). Herein, such residues are referred to as “flexible residues”. The number and position of flexible residues are not 2023229507  12 Sep 2023 particularly limited as long as the antigen-binding activity of the antigen-binding molecule of the present invention varies depending on the concentration of a tissue-specific compound. Specifically, the CDR sequences and / or FR sequences of the heavy chain and / or light chain may contain one or more flexible residues. One can identify the flexible residues and those residues 5 that can be substituted into other amino acids for library production by introducing mutations or by crystal structure analysis of complexes formed between an antibody and adenosine and / or ATP. For example, from crystal structure analysis of complexes formed between an antibody and adenosine and / or ATP, one can identify residues in the antibody that are not involved in binding to adenosine and / or ATP. One can select amino acids that can maintain binding to the 0 compounds at an appropriate level even when the residues that have been identified as not being involved in binding to adenosine and / or ATP are substituted into other amino acids. Accordingly, it is possible to design a library that has the selected amino acids for the selected residues. In this case, one can design a library mainly comprising multiple antigen-binding molecules to be an assembly of antigen-binding molecules in which residues identified as not 5 being involved in binding to adenosine and / or ATP have been substituted with amino acids that are different from one another. That is, the combination of individual flexible residues substituted with amino acids that are different from one another can provide sequence diversity in antigen-binding molecules containing the flexible residues. Antigen-binding molecules can be designed to include residues wherein at least one of 0 the residues identified to be involved in binding to adenosine and / or ATP binding becomes any residue selected from the residue and other residues that are different from the residue. In a non-limiting embodiment, examples of amino acids identified as being involved in binding to adenosine and / or ATP may include one or more amino acids selected from amino acids at positions 52, 52a, 53, 96, 100a, and 100c in the heavy chain variable region. In a non-limiting 25 embodiment, examples of such amino acids include one or more amino acids selected from amino acids including Ser at position 52, Ser at position 52a, Arg at position 53, Gly at position 96, Leu at position 100a, and Trp at position 100c contained in the heavy chain variable region. For example, when Leu at position 100a mentioned above is identified to be involved in binding to adenosine and / or ATP, the amino acid residue at position 100a in the antigen-binding 30 molecules included in the library may be any amino acid residue selected from the flexible residues of His, Met, Leu, Arg, Trp, or Tyr, in addition to Leu. In a non-limiting embodiment, examples of the flexible residues may include amino acids at positions 31, 32, 33, 35, 50, 55, 56, 57, 58, 59, 95, 96, 97, 98, 99, 100, 100a, and 100b contained in the heavy chain variable region. In another non-limiting embodiment, examples of 35 such amino acids may include amino acids at positions 26, 27, 27a, 27b, 27c, 28, 29, 31, 32, 50, 51, 52, 53, 54, 55, 89, 90, 91, 92, 93, 94, 95a, 96, and 97 contained in the light chain variable 2023229507  12 Sep 2023 region. In a non-limiting embodiment, examples of the aforementioned flexible residues may include the following amino acids contained in the heavy chain variable region: Asp, Gly, Asn, Ser, Arg, or Thr for the amino acid at position 31; 5 Ala, Phe, His, Asn, Ser, or Tyr for the amino acid at position 32; Ala, Glu, Asp, Gly, Phe, Ile, His, Lys, Met, Leu, Asn, Gln, Pro, Ser, Arg, Trp, Val, Tyr, or Thr for the amino acid at position 33; His, Ser, Thr, Tyr, or Asn for the amino acid at position 35; Ala, Glu, Asp, Gly, Phe, Ile, His, Lys, Met, Leu, Asn, Gln, Pro, Arg, Thr, Trp, Val, Tyr, or Ser for 0 the amino acid at position 50; Ala, Glu, Asp, Gly, Leu, Thr, Ser, Arg, or Asn for the amino acid at position 55; Ala, Glu, Asp, Gly, Phe, Ile, His, Lys, Met, Leu, Gln, Pro, Ser, Thr, Trp, Val, or Tyr for the amino acid at position 56; Ala, Lys, Arg, Thr, or Ile for the amino acid at position 57; 5 Asp, Gly, Phe, His, Ser, Thr, Tyr, or Asn for the amino acid at position 58; Leu, or Tyr for the amino acid at position 59; Ala, Ile, Lys, Met, Leu, Arg, Trp, Val, Tyr, or Phe for the amino acid at position 95; Ala, Asp, Asn, or Ser for the amino acid at position 96; Ala, Asp, Gly, Ile, His, Lys, Met, Leu, Asn, Ser, Val, Tyr, or Arg for the amino acid at position 0 97; Ala, Glu, Asp, Gly, Phe, Ile, His, Met, Leu, Asn, Gln, Pro, Ser, Arg, Thr, Trp, Val, Tyr, or Lys for the amino acid at position 98; Ala, Glu, Asp, Phe, His, Lys, Asn, Gln, Ser, Arg, Trp, Val, Tyr, or Gly for the amino acid at position 99; 25 Ala, Glu, Gly, Phe, Ile, His, Lys, Met, Leu, Asn, Gln, Pro, Ser, Arg, Thr, Trp, Val, Tyr, or Asp for the amino acid at position 100; Ala, Phe, Ile, His, Lys, Met, Arg, Trp, Val, or Tyr for the amino acid at position 100a; or Ala, Glu, Asp, Gly, Phe, Ile, His, Lys, Met, Leu, Gln, Pro, Ser, Arg, Thr, Trp, Val, Tyr, or Asn for the amino acid at position 100b. 30 In a non-limiting embodiment, examples of the aforementioned flexible residues may include the following amino acids contained in the light chain variable region: Ala, Ser, or Thr for the amino acid at position 26; Thr or Ser for the amino acid at position 27; Gly, Asn, Thr, or Ser for the amino acid at position 27a; 35   Asn or Asp for the amino acid at position 27b; Ile or Val for the amino acid at position 27c; 2023229507  12 Sep 2023 Asp or Gly for the amino acid at position 28; Ala, Asp, Phe, Ser, Arg, Thr, Tyr, or Gly for the amino acid at position 29; Glu, Asp, Lys, or Asn for the amino acid at position 31; Ala, Asp, Ser, Thr, or Tyr for the amino acid at position 32; 5 Asp, Gly, Lys, Asn, Gln, Ser, Arg, Tyr, or Glu for the amino acid at position 50; Asp, Gly, Lys, Asn, Thr, or Val for the amino acid at position 51; Ala, Asp, Asn, Thr, or Ser for the amino acid at position 52; Glu, Asp, His, Asn, Gln, Ser, Tyr, or Lys for the amino acid at position 53; Lys or Arg for the amino acid at position 54; 0 Leu or Pro for the amino acid at position 55; Ala, Gly, Phe, Leu, Asn, Gln, Thr, Val, Tyr, or Ser for the amino acid at position 89; Ala, Leu, Thr, Val, or Ser for the amino acid at position 90; Ala, Asp, Phe, His, Lys, Asn, Ser, Arg, Thr, Trp, Val, or Tyr for the amino acid at position 91; Glu, Asp, Ser, Arg, Thr, Val, Tyr, or Ala for the amino acid at position 92; 5 Ala, Asp, Ile, Asn, Ser, Arg, Thr, Val, Tyr, or Gly for the amino acid at position 93; Ala, Asp, Gly, Ile, Asn, Arg, Thr, or Ser for the amino acid at position 94; Ala, Glu, Asp, Gly, Phe, Ile, His, Lys, Met, Leu, Gln, Pro, Ser, Arg, Thr, Trp, Val, Tyr, or Asn for the amino acid at position 95; Ala, Glu, Asp, Gly, Ile, His, Lys, Leu, Gln, Pro, Ser, Arg, Thr, Tyr, or Asn for the amino acid at 0 position 95a; Ala, Asp, Gly, Phe, His, Lys, Leu, Asn, Gln, Pro, Ser, Thr, Trp, Tyr, or Val for the amino acid at position 96; or Ala, Gly, Ile, Met, Leu, Ser, or Val for the amino acid at position 97. Herein, “flexible residue” refers to amino acid residue variations present at 25 hypervariable amino acid positions of light-chain and heavy-chain variable regions at which several different amino acids exist, when the amino acid sequences of known and / or native antibodies or antigen-binding domains are compared. The hypervariable positions are generally located in the CDR regions. In an embodiment, the data provided by Kabat, Sequences of Proteins of Immunological Interest (National Institute of Health Bethesda Md., 1987 and 1991) 30   is useful for determining the hypervariable positions in known and / or native antibodies. Furthermore, databases on the Internet (http: / / vbase.mrc-cpe.cam.ac.uk / , and http: / / www.bioinf.org.uk / abs / index.html) provide many collected sequences of human light chains and heavy chains, and their locations. The information on the sequences and locations is useful for determining the hypervariable positions in the present invention. According to the 35 present invention, when a certain amino acid position has preferably about 2 to about 20, preferably about 3 to about 19, preferably about 4 to about 18, preferably 5 to 17, preferably 6 to 2023229507  12 Sep 2023 16, preferably 7 to 15, preferably 8 to 14, preferably 9 to 13, and preferably 10 to 12 possible amino acid residue variations, the position can be said to be hypervariable. In some embodiments, a certain amino acid position may have preferably at least about 2, preferably at least about 4, preferably at least about 6, preferably at least about 8, preferably about 10, and 5 preferably about 12 possible amino acid residue variations. A library of the present invention that contains a plurality of antigen-binding molecules having different sequences from one another can be constructed by combining heavy chain variable regions produced as a randomized variable region sequence library with the aforementioned light chain variable regions introduced with at least one amino acid residue that 0 changes the antigen-binding activity of the antigen-binding domains depending on the presence or absence of adenosine and / or ATP. Similarly, a library of the present invention that contains a plurality of antigen-binding molecules having different sequences from one another can also be produced by combining the heavy-chain variable regions introduced with at least one amino acid residue that changes the antigen-binding activity of the antigen-binding domains depending on 5 the presence or absence of adenosine and / or ATP, and having the other amino acid residues designed as flexible residues. When heavy chain variable regions produced as a randomized variable region sequence library and light chain variable regions into which at least one amino acid residue that alters the antigen-binding activity of an antigen-binding molecule depending on the concentration of the 0 target tissue-specific compound has been introduced are combined as described above, the sequences of the light chain variable regions can be designed to contain flexible residues in the same manner as described above. The number and position of such flexible residues are not particularly limited to particular embodiments as long as the antigen-binding activity of antigen-binding molecules of the present invention varies depending on the presence or absence 25 of adenosine and / or ATP. Specifically, the CDR sequences and / or FR sequences of heavy chain and / or light chain can contain one or more flexible residues. The preferred heavy chain variable regions to be combined include, for example, randomized variable region libraries. Known methods are combined as appropriate to produce a randomized variable region library. In a non-limiting embodiment of the present invention, 30 an immune library constructed based on antibody genes derived from lymphocytes of animals immunized with a specific antigen, patients with infections, persons with an elevated antibody titer in blood as a result of vaccination, cancer patients, or auto immune disease patients, may be preferably used as a randomized variable region library. In another non-limiting embodiment of the present invention, a synthetic library 35 produced by replacing the CDR sequences of V genes in genomic DNA or functional reshaped V genes with a set of synthetic oligonucleotides containing sequences encoding codon sets of an 2023229507  12 Sep 2023 appropriate length can also be preferably used as a randomized variable region library. In this case, since sequence diversity is observed in the heavy chain CDR3 sequence, it is also possible to replace the CDR3 sequence only. A criterion of giving rise to diversity in amino acids in the variable region of an antigen-binding molecule is that diversity is given to amino acid residues at 5 surface-exposed positions in the antigen-binding molecule. The surface-exposed position refers to a position that is considered to be able to be exposed on the surface and / or contacted with an antigen, based on structure, ensemble of structures, and / or modeled structure of an antigen-binding molecule. In general, such positions are CDRs. Preferably, surface-exposed positions are determined using coordinates from a three-dimensional model of an 0 antigen-binding molecule using a computer program such as the InsightII program (Accelrys). Surface-exposed positions can be determined using algorithms known in the art (for example, Lee and Richards (J. Mol. Biol. (1971) 55, 379-400); Connolly (J. Appl. Cryst. (1983) 16, 548-558)). Determination of surface-exposed positions can be performed using software suitable for protein modeling and three-dimensional structural information obtained from an 5 antibody. Software that can be used for these purposes preferably includes SYBYL Biopolymer Module software (Tripos Associates). Generally or preferably, when an algorithm requires a user input size parameter, the "size" of a probe which is used in the calculation is set at about 1.4 Angstrom or smaller in radius. Furthermore, methods for determining surface-exposed regions and areas using software for personal computers are described by Pacios (Comput. Chem. (1994) 0 18 (4), 377-386; J. Mol. Model. (1995) 1, 46-53). Furthermore, in a non-limiting embodiment of the present invention, amino acids of the variable region including the CDR region and / or the framework region may be altered appropriately to improve antibody stability. In a non-limiting embodiment, examples of such amino acids may include the amino acids of positions 1, 5, 10, 30, 48, and 58. More 25 specifically, examples may include Gln at position 1, Gln at position 5, Asp at position 10, Asn at position 30, Leu at position 48, and Asn at position 58. For the improvement of antibody stability, these amino acids can be substituted for corresponding amino acids contained in a germ-line sequence. In a non-limiting embodiment, an example of such a germ line sequence may be the VH3-21 sequence. In this case, Gln of position 1 may be substituted with Glu, Gln 30 of position 5 may be substituted with Val, Asp of position 10 may be substituted with Gly, Asn of position 30 may be substituted with Ser, Leu of position 48 may be substituted with Val, and Asn of position 58 may be substituted with Tyr. In another non-limiting embodiment of the present invention, a naive library which is constructed from antibody genes derived from lymphocytes of healthy individuals and consists 35 of naive sequences which are antibody sequences that do not have bias in their repertoire, can also be particularly preferably used as a randomized variable region library (Gejima et al. 2023229507  12 Sep 2023 (Human Antibodies (2002) 11, 121-129); Cardoso et al. (Scand. J. Immunol. (2000) 51, 337-344)). Herein, “an amino acid sequence comprising a naive sequence” refers to an amino acid sequence obtained from such a naive library. 5 Fc region An Fc region contains an amino acid sequence derived from the heavy chain constant region of an antibody. An Fc region is a portion of the antibody heavy chain constant region that includes the N terminal end of the hinge region, which is the papain cleavage site, at an amino acid around position 216 (indicated by EU numbering), and the hinge, CH2, and CH3 0 domains. Fc regions can be obtained from human IgG1; however, they are not limited to any specific IgG subclass. Preferred examples of the Fc regions include Fc regions having FcRn-binding activity in an acidic pH range as described below. Preferred examples of the Fc regions include Fc regions having Fcy receptor-binding activity as described below. In a non-limiting embodiment, examples of such Fc regions include the Fc regions of human IgG1 5 (SEQ ID NO: 5), IgG2 (SEQ ID NO: 6), IgG3 (SEQ ID NO: 7), or IgG4 (SEQ ID NO: 8). Fcy receptor (FcyR) “FcY receptor” (also called “FcyR”) refers to a receptor capable of binding to the Fc region of monoclonal IgG1, IgG2, IgG3, or IgG4 antibodies; and means all members belonging 0 to the family of proteins substantially encoded by Fcy receptor genes. In humans, the family includes FcyRI (CD64) including isoforms FcyRIa, FcyRIb, and FcyRIc; FcyRII (CD32) including isoforms FcyRIIa (including allotype H131 and R131, i.e., FcyRIIa(H) and FcyRIIa(R)), FcyRIIb (including FcyRIIb-1 and FcyRIIb-2), and FcyRIIc; and FcyRIII (CD16) including isoform FcyRIIIa (including allotype V158 and F158, i.e., FcyRIIIa(V) and 25   FcyRIIIa(F)) and FcyRIIIb (including allotype FcyRIIIb-NA1 and FcyRIIIb-NA2); as well as all unidentified human FcyRs, FcyR isoforms, and allotypes thereof; but the family is not limited to these examples. Without being limited thereto, FcyRs include those derived from humans, mice, rats, rabbits, and monkeys. FcyRs may be derived from any organism. Mouse FcyRs include FcyRI (CD64), FcyRII (CD32), FcyRIII (CD16), and FcyRIII-2 (FcyRIV, CD16-2), as well as all 30   unidentified mouse FcyRs, FcyR isoforms, and allotypes thereof, but they are not limited to these examples. Preferred examples of such Fcy receptors include, human FcyRI (CD64), FcyRIIa (CD32), FcyRIIb (CD32), FcyRIIIa (CD16), and / or FcyRIIIb (CD16). The polynucleotide sequence and amino acid sequence of human FcyRI are shown in SEQ ID NOs: 9 (NM_000566.3) and 10 (NP_000557.1), respectively; the polynucleotide sequence and amino 35 acid sequence of human FcyRIIa (allotype H131) are shown in SEQ ID NOs: 11 (BC020823.1) and 12 (AAH20823.1), respectively (allotype R131 is a sequence in which the amino acid at 2023229507  12 Sep 2023 position 166 of SEQ ID NO: 12 is substituted with Arg); the polynucleotide sequence and amino acid sequence of FcYlIb are shown in SEQ ID NOs: 13 (BC146678.1) and 14 (AAI46679.1), respectively; the polynucleotide sequence and amino acid sequence of FcyRIIIa are shown in SEQ ID NOs: 15 (BC033678.1) and 16 (AAH33678.1), respectively; and the polynucleotide 5 sequence and amino acid sequence of FcYRIIIb are shown in SEQ ID NOs: 17 (BC128562.1) and 18 (AAI28563.1), respectively (RefSeq accession number or such is shown in parentheses). Whether an Fcy receptor has binding activity to the Fc region of a monoclonal IgG1, IgG2, IgG3, or IgG4 antibody can be assessed by ALPHA (Amplified Luminescent Proximity Homogeneous Assay) screen, surface plasmon resonance (SPR)-based BIACORE methods, and others (Proc. 0 Natl. Acad. Sci. USA (2006) 103(11), 4005-4010), in addition to the above-described FACS and ELISA formats. In FcyRI (CD64) including FcyRIa, FcYRIb, and FcyRIc, and FcyRIII (CD16) including isoforms FcYRIIIa (including allotypes V158 and F158) and FcYRIIIb (including allotypes FcYRIIIb-NA1 and FcYRIIIb-NA2), a chain that binds to the Fc region of IgG is associated with 5   common Y chain having ITAM responsible for transduction of intracellular activation signal. Meanwhile, the cytoplasmic domain of FcYRII (CD32) including isoforms FcYRIIa (including allotypes H131 and R131) and FcYRIIc contains ITAM. These receptors are expressed on many immune cells such as macrophages, mast cells, and antigen-presenting cells. The activation signal transduced upon binding of these receptors to the Fc region of IgG results in enhancement 0   of the phagocytic activity of macrophages, inflammatory cytokine production, mast cell degranulation, and the enhanced function of antigen-presenting cells. FcY receptors having the ability to transduce the activation signal as described above are herein referred to as activating FcY receptors. Meanwhile, the intracytoplasmic domain of FcYRIIb (including FcYRIIb-1 and 25   FcYRIIb-2) contains ITIM responsible for transduction of inhibitory signals. The crosslinking between FcYRIIb and B cell receptor (BCR) on B cells suppresses the activation signal from BCR, which results in suppression of antibody production via BCR. The crosslinking of FcYRIII and FcYRIIb on macrophages suppresses the phagocytic activity and inflammatory cytokine production. FcY receptors having the ability to transduce the inhibitory signal as 30 described above are herein referred to as inhibitory FcY receptor. FcYR-binding activity of Fc region As mentioned above, Fc regions having an FcY receptor-binding activity are examples of Fc regions comprised in the antigen-binding molecules of the present invention. A non-limiting 35 embodiment of such an Fc region includes the Fc region of human IgG1 (SEQ ID NO: 5), IgG2 (SEQ ID NO: 6), IgG3 (SEQ ID NO: 7), or IgG4 (SEQ ID NO: 8). Whether an FcY receptor 2023229507  12 Sep 2023 has binding activity to the Fc region of a monoclonal IgG1, IgG2, IgG3, or IgG4 antibody can be assessed by ALPHA screen (Amplified Luminescent Proximity Homogeneous Assay), surface plasmon resonance (SPR)-based BIACORE method, and others (Proc. Natl. Acad. Sci. U.S.A. (2006) 103(11), 4005-4010), in addition to the above-described FACS and ELISA formats. 5 ALPHA screen is performed by the ALPHA technology based on the principle described below using two types of beads: donor and acceptor beads. A luminescent signal is detected only when molecules linked to the donor beads interact biologically with molecules linked to the acceptor beads and when the two beads are located in close proximity. Excited by laser beam, the photosensitizer in a donor bead converts oxygen around the bead into excited singlet oxygen. 0 When the singlet oxygen diffuses around the donor beads and reaches the acceptor beads located in close proximity, a chemiluminescent reaction within the acceptor beads is induced. This reaction ultimately results in light emission. If molecules linked to the donor beads do not interact with molecules linked to the acceptor beads, the singlet oxygen produced by donor beads do not reach the acceptor beads and chemiluminescent reaction does not occur. 5 For example, a biotin-labeled antigen-binding molecule comprising Fc region is immobilized to the donor beads and glutathione S-transferase (GST)-tagged Fcy receptor is immobilized to the acceptor beads. In the absence of an antigen-binding molecule comprising a competitive Fc region variant, Fcy receptor interacts with an antigen-binding molecule comprising a native Fc region, inducing a signal of 520 to 620 nm as a result. The 0   antigen-binding molecule having a non-tagged Fc region variant competes with the antigen-binding molecule comprising a native Fc region for the interaction with Fcy receptor. The relative binding affinity can be determined by quantifying the reduction of fluorescence as a result of competition. Methods for biotinylating the antigen-binding molecules such as antibodies using Sulfo-NHS-biotin or the like are known. Appropriate methods for adding the 25 GST tag to an Fcy receptor include methods that involve fusing polypeptides encoding Fcy and GST in-frame, expressing the fused gene using cells introduced with a vector to which the gene is operablye linked, and then purifying using a glutathione column. The induced signal can be preferably analyzed, for example, by fitting to a one-site competition model based on nonlinear regression analysis using software such as GRAPHPAD PRISM (GraphPad; San Diego). 30 One of the substances for observing their interaction is immobilized as a ligand onto the gold thin layer of a sensor chip. When light is shed on the rear surface of the sensor chip so that total reflection occurs at the interface between the gold thin layer and glass, the intensity of reflected light is partially reduced at a certain site (SPR signal). The other substance for observing their interaction is injected as an analyte onto the surface of the sensor chip. The 35 mass of immobilized ligand molecule increases when the analyte binds to the ligand. This alters the refraction index of solvent on the surface of the sensor chip. The change in refraction 2023229507  12 Sep 2023 index causes a positional shift of SPR signal (conversely, the dissociation shifts the signal back to the original position). In the Biacore system, the amount of shift described above (i.e., the change of mass on the sensor chip surface) is plotted on the vertical axis, and thus the change of mass over time is shown as measured data (sensorgram). Kinetic parameters (association rate 5   constant (ka) and dissociation rate constant (kd)) are determined from the curve of sensorgram, and affinity (KD) is determined from the ratio between these constants. Inhibition assay is preferably used in the BIACORE methods. Examples of such inhibition assay are described in Proc. Natl. Acad. Sci. U.S.A. (2006) 103(11), 4005-4010. 0 Fc regions with altered Fcy receptor (FcyR) binding In addition to the Fc region of human IgG1 (SEQ ID NO: 5), IgG2 (SEQ ID NO: 6), IgG3 (SEQ ID NO: 7), or IgG4 (SEQ ID NO: 8), an Fc region with altered FcyR binding, which has a higher Fcy receptor-binding activity than an Fc region of a native human IgG may be appropriately used as an Fc region included in the present invention. Herein, “Fc region of a 5 native human IgG” refers to an Fc region in which the sugar chain bonded to position 297 (EU numbering) of the Fc region of human IgG1, IgG2, IgG3, or IgG4 shown in SEQ ID NOs: 5, 6, 7, or 8 is a fucose-containing sugar chain. Such Fc regions with altered FcyR binding may be produced by altering amino acids of the Fc region of a native human IgG. Whether the FcyR-binding activity of an Fc region with altered FcyR binding is higher than that of an Fc 0 region of a native human IgG can be determined appropriately using methods described in the abovementioned section on binding activity. In the present invention, "alteration of amino acids" or "amino acid alteration" of an Fc region includes alteration into an amino acid sequence which is different from that of the starting Fc region. The starting Fc region may be any Fc region, as long as a variant modified from the 25 starting Fc region can bind to human Fcy receptor in a neutral pH range. Furthermore, an Fc region altered from a starting Fc region which had been already altered can also be used preferably as an Fc region of the present invention. The “starting Fc region” can refer to the polypeptide itself, a composition comprising the starting Fc region, or an amino acid sequence encoding the starting Fc region. Starting Fc regions can comprise known Fc regions produced 30 via recombination described briefly in the section “Antibodies”. The origin of starting Fc regions is not limited, and they may be obtained from human or any nonhuman organisms. Such organisms preferably include mice, rats, guinea pigs, hamsters, gerbils, cats, rabbits, dogs, goats, sheep, bovines, horses, camels and organisms selected from nonhuman primates. In another embodiment, starting Fc regions can also be obtained from cynomolgus monkeys, 35 marmosets, rhesus monkeys, chimpanzees, or humans. Starting Fc regions can be obtained preferably from human IgG1; however, they are not limited to any particular IgG class. This 2023229507  12 Sep 2023 means that an Fc region of human IgG1, IgG2, IgG3, or IgG4 can be used appropriately as a starting Fc region, and herein also means that an Fc region of an arbitrary IgG class or subclass derived from any organisms described above can be preferably used as a starting Fc region. Examples of native IgG variants or altered forms are described in published documents (Curr. 5 Opin. Biotechnol. (2009) 20 (6): 685-91; Curr. Opin. Immunol. (2008) 20 (4), 460-470; Protein Eng. Des. Sel. (2010) 23 (4): 195-202; International Publication Nos. WO 2009 / 086320, WO 2008 / 092117, WO 2007 / 041635, and WO 2006 / 105338); however, they are not limited to the examples. Examples of alterations include those with one or more mutations, for example, 0 mutations by substitution of different amino acid residues for amino acids of starting Fc regions, by insertion of one or more amino acid residues into starting Fc regions, or by deletion of one or more amino acids from starting Fc region. Preferably, the amino acid sequences of altered Fc regions comprise at least a part of the amino acid sequence of a non-native Fc region. Such variants necessarily have sequence identity or similarity less than 100% to their starting Fc 5 region. In a preferred embodiment, the variants have amino acid sequence identity or similarity about 75% to less than 100%, more preferably about 80% to less than 100%, even more preferably about 85% to less than 100%, still more preferably about 90% to less than 100%, and yet more preferably about 95% to less than 100% to the amino acid sequence of their starting Fc region. In a non-limiting embodiment of the present invention, at least one amino acid is 0 different between an FcYR-binding altered Fc region of the present invention and its starting Fc region. Amino acid difference between an FcYR-binding altered Fc region of the present invention and its starting Fc region can also be preferably specified based on the specific amino acid differences at the above-described specific amino acid positions by EU numbering. Examples of methods of preparing such variants are shown in the section “Alteration of amino 25 acids”. Included in the antigen-binding molecules of the present invention, an Fc region with altered FcYR binding, which has a higher FcY receptor-binding activity than that of an Fc region of a native human IgG, (an FcYR binding-altered Fc region) may be obtained by any method. Specifically, the Fc region with altered FcYR binding may be obtained by altering amino acids of 30 an IgG-type human immunoglobulin used as a starting Fc region. Preferred Fc regions of the IgG-type immunoglobulins for alteration include, for example, those of human IgGs shown in SEQ ID NOs: 5, 6, 7, or 8 (IgG1, IgG2, IgG3, or IgG4, respectively, and variants thereof). Amino acids of any positions may be altered into other amino acids, as long as the binding activity toward the FcY receptor is higher than that of the Fc region of a native human 35 IgG. When the antigen-binding molecule contains a human IgG1 Fc region as the human Fc region, it preferably contains an alteration that yields the effect of a higher FcY receptor-binding 2023229507  12 Sep 2023 activity than that of the Fc region of a native human IgG, in which the sugar chain bound at position 297 (EU numbering) is a fucose-containing sugar chain. Such amino acid alterations have been reported, for example, in international publications such as WO2007 / 024249, WO2007 / 021841, WO2006 / 031370, WO2000 / 042072, WO2004 / 029207, WO2004 / 099249, 5 WO2006 / 105338, WO2007 / 041635, WO2008 / 092117, WO2005 / 070963, WO2006 / 020114, WO2006 / 116260, and WO2006 / 023403. Examples of such amino acids that may be altered include at least one or more amino acids selected from the group consisting of positions 221, 222, 223, 224, 225, 227, 228, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 246, 247, 249, 250, 251, 254, 0 255, 256, 258, 260, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 278, 279, 280, 281, 282, 283, 284, 285, 286, 288, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 311, 313, 315, 317, 318, 320, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 339, 376, 377, 378, 379, 380, 382, 385, 392, 396, 421, 427, 428, 429, 434, 436, and 440 (EU numbering). An Fc region (Fc region with 5   altered FcyR binding) having a higher Fcy receptor-binding activity than that of an Fc region of a native human IgG can be obtained by altering these amino acids. Examples of particularly preferable alterations for use in the present invention include at least one or more amino acid alterations selected from the group consisting of: Lys or Tyr for the amino acid of position 221; 0   Phe, Trp, Glu, or Tyr for the amino acid of position 222; Phe, Trp, Glu, or Lys for the amino acid of position 223; Phe, Trp, Glu, or Tyr for the amino acid of position 224; Glu, Lys, or Trp for the amino acid of position 225; Glu, Gly, Lys, or Tyr for the amino acid of position 227; 25   Glu, Gly, Lys, or Tyr for the amino acid of position 228; Ala, Glu, Gly, or Tyr for the amino acid of position 230; Glu, Gly, Lys, Pro, or Tyr for the amino acid of position 231; Glu, Gly, Lys, or Tyr for the amino acid of position 232; Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the 30 amino acid of position 233; Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 234; Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 235; 35 Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 236; 2023229507  12 Sep 2023 Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 237; Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 238; 5 Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr for the amino acid of position 239; Ala, Ile, Met, or Thr for the amino acid of position 240; Asp, Glu, Leu, Arg, Trp, or Tyr for the amino acid of position 241; Leu, Glu, Leu, Gln, Arg, Trp, or Tyr for the amino acid of position 243; 0 His for the amino acid of position 244; Ala for the amino acid of position 245; Asp, Glu, His, or Tyr for the amino acid of position 246; Ala, Phe, Gly, His, Ile, Leu, Met, Thr, Val, or Tyr for the amino acid of position 247; Glu, His, Gln, or Tyr for the amino acid of position 249; 5 Glu or Gln for the amino acid of position 250; Phe for the amino acid of position 251; Phe, Met, or Tyr for the amino acid of position 254; Glu, Leu, or Tyr for the amino acid of position 255; Ala, Met, or Pro for the amino acid of position 256; 0 Asp, Glu, His, Ser, or Tyr for the amino acid of position 258; Asp, Glu, His, or Tyr for the amino acid of position 260; Ala, Glu, Phe, Ile, or Thr for the amino acid of position 262; Ala, Ile, Met, or Thr for the amino acid of position 263; Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Trp, or Tyr for the 25 amino acid of position 264; Ala, Leu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 265; Ala, Ile, Met, or Thr for the amino acid of position 266; Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Thr, Val, Trp, or Tyr for the amino 30 acid of position 267; Asp, Glu, Phe, Gly, Ile, Lys, Leu, Met, Pro, Gln, Arg, Thr, Val, or Trp for the amino acid of position 268; Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 269; 35 Glu, Phe, Gly, His, Ile, Leu, Met, Pro, Gln, Arg, Ser, Thr, Trp, or Tyr for the amino acid of position 270; 2023229507  12 Sep 2023 Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 271; Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 272; 5 Phe or Ile for the amino acid of position 273; Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 274; Leu or Trp for the amino acid of position 275; Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of 0 position 276; Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, or Trp for the amino acid of position 278; Ala for the amino acid of position 279; Ala, Gly, His, Lys, Leu, Pro, Gln, Trp, or Tyr for the amino acid of position 280; 5 Asp, Lys, Pro, or Tyr for the amino acid of position 281; Glu, Gly, Lys, Pro, or Tyr for the amino acid of position 282; Ala, Gly, His, Ile, Lys, Leu, Met, Pro, Arg, or Tyr for the amino acid of position 283; Asp, Glu, Leu, Asn, Thr, or Tyr for the amino acid of position 284; Asp, Glu, Lys, Gln, Trp, or Tyr for the amino acid of position 285; 0 Glu, Gly, Pro, or Tyr for the amino acid of position 286; Asn, Asp, Glu, or Tyr for the amino acid of position 288; Asp, Gly, His, Leu, Asn, Ser, Thr, Trp, or Tyr for the amino acid of position 290; Asp, Glu, Gly, His, Ile, Gln, or Thr for the amino acid of position 291; Ala, Asp, Glu, Pro, Thr, or Tyr for the amino acid of position 292; 25 Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 293; Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 294; Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid 30 of position 295; Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, or Val for the amino acid of position 296; Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 297; 35 Ala, Asp, Glu, Phe, His, Ile, Lys, Met, Asn, Gln, Arg, Thr, Val, Trp, or Tyr for the amino acid of position 298; 2023229507  12 Sep 2023 Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Val, Trp, or Tyr for the amino acid of position 299; Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, or Trp for the amino acid of position 300; 5 Asp, Glu, His, or Tyr for the amino acid of position 301; Ile for the amino acid of position 302; Asp, Gly, or Tyr for the amino acid of position 303; Asp, His, Leu, Asn, or Thr for the amino acid of position 304; Glu, Ile, Thr, or Tyr for the amino acid of position 305; 0 Ala, Asp, Asn, Thr, Val, or Tyr for the amino acid of position 311; Phe for the amino acid of position 313; Leu for the amino acid of position 315; Glu or Gln for the amino acid of position 317; His, Leu, Asn, Pro, Gln, Arg, Thr, Val, or Tyr for the amino acid of position 318; 5 Asp, Phe, Gly, His, Ile, Leu, Asn, Pro, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 320; Ala, Asp, Phe, Gly, His, Ile, Pro, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 322; Ile for the amino acid of position 323; Asp, Phe, Gly, His, Ile, Leu, Met, Pro, Arg, Thr, Val, Trp, or Tyr for the amino acid of position 0 324; Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 325; Ala, Asp, Glu, Gly, Ile, Leu, Met, Asn, Pro, Gln, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 326; 25 Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg, Thr, Val, Trp, or Tyr for the amino acid of position 327; Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 328; Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the 30 amino acid of position 329; Cys, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg, Ser, Thr, Val, Trp, or Tyr for the amino acid of position 330; Asp, Phe, His, Ile, Leu, Met, Gln, Arg, Thr, Val, Trp, or Tyr for the amino acid of position 331; Ala, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp, or Tyr for the 35 amino acid of position 332; Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Pro, Ser, Thr, Val, or Tyr for the amino acid of 2023229507  12 Sep 2023 position 333; Ala, Glu, Phe, Ile, Leu, Pro, or Thr for the amino acid of position 334; Asp, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Arg, Ser, Val, Trp, or Tyr for the amino acid of position 335; 5 Glu, Lys, or Tyr for the amino acid of position 336; Glu, His, or Asn for the amino acid of position 337; Asp, Phe, Gly, Ile, Lys, Met, Asn, Gln, Arg, Ser, or Thr for the amino acid of position 339; Ala or Val for the amino acid of position 376; Gly or Lys for the amino acid of position 377; 0 Asp for the amino acid of position 378; Asn for the amino acid of position 379; Ala, Asn, or Ser for the amino acid of position 380; Ala or Ile for the amino acid of position 382; Glu for the amino acid of position 385; 5 Thr for the amino acid of position 392; Leu for the amino acid of position 396; Lys for the amino acid of position 421; Asn for the amino acid of position 427; Phe or Leu for the amino acid of position 428; 0 Met for the amino acid of position 429; Trp for the amino acid of position 434; Ile for the amino acid of position 436; and Gly, His, Ile, Leu, or Tyr for the amino acid of position 440; as indicated by EU numbering in the Fc region. The number of amino acids to be altered is not 25 particularly limited; and amino acid may be altered at only one site or amino acids may be altered at two or more sites. Examples of combinations for amino acid alterations at two or more sites include those described in Table 1 (Tables 1-1 to 1-3). 2023229507  12 Sep 2023 Table 1-1 Combination of amino acids Combination of amino acids K370E / P396L / D270E S239Q / I332Q Q419H / P396L / D270E S267D / I332E V240A / P396L / D270E S267E / I332E R255L / P396L / D270E S267L / A327S R255L / P396L / D270E R255L / P396L / D270E / R292G R255L / P396L / D270E R255L / P396L / D270E / Y300L F243L / D270E / K392N / P396L F243L / R255L / D270E / P396L F243L / R292P / Y300L / V305I / P396L F243L / R292P / Y300L / P396L F243L / R292P / Y300L F243L / R292P / P396L F243L / R292P / V305I F243L / R292P _____ ___ S298A / E333A / K334A E380A / T307A K326M / E333S K326A / E333A S317A / K353A A327D / I332E A330L / I332E A330Y / I332E E258H / I332E E272H / I332E E272I / N276D S267Q / A327S S298A / I332E S304T / 1332E S324G / I332D S324G / I332E S324I / I332D S324I / I332E T260H / I332E T335D / I332E V240I / V266I V264I / I332E D265F / N297E / I332E D265Y / N297D / I332E F243L / V262I / V264W N297D / A330Y / I332E N297D / T299E / I332E N297D / T299F / I332E -----   -     — -------------- ------------------—    ----------- ------- ---- -     — —   N297D / T299H / 1332E N297D / T299I / 1332E N297D / T299L / 1332E N297D / T299V / I332E P230A / E233D / 1332E P244H / P245A / P247V E272R / 1332E S239D / A330L / 1332E E283H / I332E S239D / A330Y / 1332E E293R / 1332E S239D / H268E / A330Y F241L / V2621 S239D / 1332E / A327A F241W / F243W S239D / 1332E / A3301 Table 1-2 is a continuation of Table 1-1. 2023229507  12 Sep 2023 Table 1-2 F243L / V264I S239D / N297D / I332E H268D / A330Y S239D / S298A / I332E H268E / A330Y S239D / V264I / I332E K246H / I332E S239E / N297D / 1332E L234D / I332E L234E / 1332E L234G / I332E L234I / I332E L234I / L235D L234Y / I332E L235D / I332E L235E / 1332E L235I / 1332E L235S / I332E L328A / I332D L328D / I332D L328D / 1332E L328E / I332D L328E / I332E L328F / 1332D L328F / 1332E L328H / 1332E L328I / I332D L328I / I332E L328M / I332D L328M / I332E L328N / I332D L328N / I332E L328Q / I332D L328Q / 1332E L328T / 1332D S239E / V264I / I332E S239N / A330L / I332E S239N / A330Y / I332E S239N / S298A / I332E S239Q / V264I / I332E V264E / N297D / I332E V2641 / A330L / 1332E V2641 / A330Y / I332E V264I / S298A / I332E Y296D / N297D / I332E Y296E / N297D / 1332E Y296H / N297D / I332E Y296N / N297D / I332E Y296Q / N297D / 1332E Y296T / N297D / I332E D265Y / N297D / T299L / I332E F241E / F243Q / V262T / V264E F241E / F243R / V262E / V264R F241E / F243Y / V262T / V264R F241 L / F243L / V262I / V264I   __________ F241R / F243Q / V262T / V264R F241S / F243H / V262T / V264T F241U1 / F243^ / V262A / V264A F241Y / F243Y / V262T / V264T 1332E / A330Y / H268E / A327A N297D / I332E / S239D / A330L N297D / S298A / A330Y / 1332E L328T / 1332E S239D / A330Y / 1332E / K326E L328V / I332D S239D / A330Y / 1332E / K326T L328V / I332E S239D / A330Y / 1332E / L234I L328Y / 1332D S239D / A330Y / I332E / L235D 2023229507  12 Sep 2023 Table 1-3 is a continuation of Table 1-2. Table 1-3 L328Y / 1332E S239D / A330Y / I332E / V240I N297D / I332E S239D / A330Y / 1332E / V264T N297E / I332E S239D / A330Y / I332E / V266I N297S / I332E S239D / D265F / N297D / I332E P227G / I332E S239D / D265H / N297D / I332E P230A / E233D S239D / D265I / N297D / I332E Q295E / I332E S239D / D265L / N297D / I332E R255Y / I332E S239D / D265T / N297D / 1332E S239D / I332D S239D / D265V / N297D / I332E S239D / I332E S239D / D265Y / N297D / I332E S239D / I332N S239D / I332E / A330Y / A327A S239D / I332Q S239D / I332E / H268E / A327A g39E / l)2f>SC, S239D / I332E / H268E / A330Y S239E / D265N S239D / N297D / I332E / A330Y S239E / D265Q S239D / N297D / I332E / K326E S239E / I332D S239D / N297D / I332E / L235D S239E / 1332E S239D / V264I / A330L / I332E S239E / I332N S239D / V264I / S298A / I332E S239E / I332Q S239E / V264I / A330Y / 1332E S239N / 1332D F241E / F243Q / V262T / V264E / I332E S239N / I332E F241E / F243R / V262E / V264R / I332E S239N / 1332N F241E / F243Y / V262T / V264R / I332E S239N / I332Q F241 R / F243Q / V262T / V264R / I332E S239Q / I332D S239D / I332E / H268E / A330Y / A327A S239Q / I332E S239E / V264I / S298A / A330Y / I332E S239Q / I332N F24] Y / F243Y / V262T / V264T / N297D / I332E S267E / L328F G236D / S267E S239D / S267E 5 For the pH conditions to measure the binding activity of the Fcy receptor binding domain and the Fcy receptor contained in the antigen-binding molecule of the present invention, conditions in an acidic pH range or in a neutral pH range may be suitably used. The acidic pH range or neutral pH range, as a condition to measure the binding activity of the Fcy receptor 2023229507  12 Sep 2023 binding domain and the Fcy receptor contained in the antigen-binding molecule of the present invention, generally indicates pH 5.8 to pH 8.0. Preferably, it is a range indicated with arbitrary pH values between pH 6.0 and pH 7.4; and preferably, it is selected from pH 6.0, pH 6.1, pH 6.2, pH 6.3, pH 6.4, pH 6.5, pH 6.6, pH 6.7, pH 6.8, pH 6.9, pH 7.0, pH 7.1, pH 7.2, pH 7.3, and pH 5 7.4; and particularly preferably, it is pH 6.15 to 7.4, which is close to the pH of cancer tissues (Vaupel et al., Cancer Res. (1989) 49, 6449-6665). With regard to the temperature used as a measurement condition, the binding affinity between an Fcy receptor binding domain and a human Fcy receptor can be evaluated at any temperature between 10°C and 50°C. Preferably, a temperature between 15°C and 40°C is used to determine the binding affinity between a human 0   Fcy receptor binding domain and Fcy receptor. More preferably, any temperature between 20°C and 35°C, such as any single temperature from 20°C, 21°C, 22°C, 23°C, 24°C, 25°C, 26°C, 27°C, 28°C, 29°C, 30°C, 31°C, 32°C, 33°C, 34°C, and 35°C, can be similarly used to determine the binding affinity between an Fcy receptor binding domain and an Fcy receptor. A temperature of 25°C is a non-limiting example in an embodiment of the present invention. 5 Herein, “Fc region with altered FcyR binding has a higher Fcy receptor-binding activity than the native Fc region” means that the human Fcy receptor-binding activity of the Fc region with altered FcyR binding toward any of the human Fcy receptors of FcyRI, FcyRIIa, FcyRIIb, FcyRIIIa, and / or FcyRIIIb is higher than the binding activity of the native Fc region toward these human Fcy receptors. For example, it means that based on an above-described analytical 0 method, in comparison to the binding activity of an antigen-binding molecule containing a native human IgG Fc region as a control, the binding activity of the antigen-binding molecule comprising an Fc region with altered FcyR binding is 105% or more, preferably 110% or more, 115% or more, 120% or more, 125% or more, particularly preferably 130% or more, 135% or more, 140% or more, 145% or more, 150% or more, 155% or more, 160% or more, 165% or 25 more, 170% or more, 175% or more, 180% or more, 185% or more, 190% or more, 195% or more, 2-fold or more, 2.5-fold or more, 3-fold or more, 3.5-fold or more, 4-fold or more, 4.5-fold or more, 5-fold or more, 7.5-fold or more, 10-fold or more, 20-fold or more, 30-fold or more, 40-fold or more, 50-fold or more, 60-fold or more, 70-fold or more, 80-fold or more, 90-fold or more, or 100-fold or more. The starting Fc region may be used as a native Fc region, and 30 native Fc regions of antibodies of the same subclass may also be used. In the present invention, an Fc region of a native human IgG in which the sugar chain bonded to the amino acid at position 297 (EU numbering) is a fucose-containing sugar chain, is suitably used as a native Fc region of human IgG to be used as a control. Whether or not the sugar chain bonded to the amino acid at position 297 (EU numbering) is a fucose-containing 35 sugar chain can be determined using the technique described in Non-Patent Document 6. For example, it is possible to determine whether or not the sugar chain bonded to the native human 2023229507  12 Sep 2023 IgG Fc region is a fucose-containing sugar chain by a method such as the one below. Sugar chain is dissociated from a native human IgG to be tested, by reacting the test native human IgG with N-Glycosidase F (Roche diagnostics) (Weitzhandler et al. (J. Pharma. Sciences (1994) 83, 12, 1670-1675)). Next, a dried concentrate of a reaction solution from which protein has been 5 removed by reaction with ethanol (Schenk et al. (J. Clin. Investigation (2001) 108 (11) 1687-1695)) is fluorescently labeled with 2-aminopyridine (Bigge et al. (Anal. Biochem. (1995) 230 (2) 229-238)). Reagents are removed by solid extraction using a cellulose cartridge, and the fluorescently labeled 2-AB-modified sugar chain is analyzed by normal-phase chromatography. It is possible to determine whether or not the sugar chain bonded to the native 0 Fc region of a human IgG is a fucose-containing sugar chain by observing the detected chromatogram peaks. As an antigen-binding molecule containing a native Fc region of an antibody of the same subclass, which is to be used as a control, an antigen-binding molecule having an Fc region of a monoclonal IgG antibody may be suitably used. The structures of the Fc regions are 5 described in SEQ ID NO: 5 (A is added to the N terminus of Database Accession No. AAC82527.1), SEQ ID NO: 6 (A is added to the N terminus of Database Accession No. AAB59393.1), SEQ ID NO: 7 (Database Accession No. CAA27268.1), and SEQ ID NO: 8 (A is added to the N terminus of Database Accession No. AAB59394.1). Further, when an antigen-binding molecule containing an Fc region of a particular antibody isotype is used as the 0 test substance, the effect of the antigen-binding molecule containing the test Fc region on Fcy receptor-binding activity is tested by using as a control an antigen-binding molecule having an Fc region of a monoclonal IgG antibody of that particular isotype. In this way, antigen-binding molecules containing an Fc region of which Fcy receptor-binding activity is demonstrated to be high are suitably selected. 25 Fc regions having a selective binding activity toward an Fcy receptor Examples of Fcy receptor binding domains suitable for use in the present invention include Fcy receptor binding domains having a higher binding activity to a particular Fcy receptor than to other Fcy receptors (Fcy receptor binding domains having a selective binding 30 activity to an Fcy receptor). When an antibody is used as the antigen-binding molecule (when an Fc region is used as the Fcy receptor binding domain), a single antibody molecule can only bind to a single Fcy receptor molecule. Therefore, a single antigen-binding molecule cannot bind to other activating FcyRs in an inhibitory Fcy receptor-bound state, and cannot bind to other activating Fcy receptors or inhibitory Fcy receptors in an activating Fcy receptor-bound state. 35 Fc regions with a higher binding activity toward an activating Fcy receptor than the binding 2023229507  12 Sep 2023 activity toward an inhibitory Fcy receptor As described above, preferable activating Fcy receptors include FcyRI (CD64) including FcYRIa, FcyRIb, and FcyRIc; FcYRIIa; and FcyRIII (CD16) including FcYRIIIa (including allotypes V158 and F158) and FcyRIIIb (including allotypes FcyRIIIb-NA1 and FcyRIIIb-NA2). 5   Meanwhile, preferred examples of inhibitory Fcy receptors include FcyRIIb (including FcyRIIb-1 and FcyRIIb-2). Herein, an example of a case where the binding activity toward a certain Fcy receptor is higher than the binding activity toward another Fcy receptor is the case where the binding activity toward an activating Fcy receptor is higher than the binding activity toward an inhibitory 0   Fcy receptor. In this case, the binding activity of the Fc region toward any of the human Fcy receptors of FcyRIa, FcyRIIa, FcyRIIIa, and / or FcyRIIIb is said to be higher than the binding activity toward FcyRIIb. For example, this means that, based on an above-described analytical method, the binding activity of an antigen-binding molecule containing the Fc region toward any of the human Fcy receptors, FcyRIa, FcyRIIa, FcyRIIIa, and / or FcyRIIIb, is 105% or more, 5 preferably 110% or more, 120% or more, 130% or more, 140% or more, particularly preferably 150% or more, 160% or more, 170% or more, 180% or more, 190% or more, 200% or more, 250% or more, 300% or more, 350% or more, 400% or more, 450% or more, 500% or more, 750% or more, 10-fold or more, 20-fold or more, 30-fold or more, 40-fold or more, 50-fold or more, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold or more as compared with the binding 0   activity toward FcyRIIb. The Fc region with a higher binding activity toward activating Fcy receptors than to inhibitory Fcy receptors may be favorably included in antigen-binding molecules of the present invention whose antigen-binding domain binds to a membrane-type molecule. IgG1 antibodies containing such Fc regions are known to enhance the ADCC activity mentioned below. Therefore, antigen-binding molecules containing the Fc-region are 25 also useful as antigen-binding molecules to be included in the pharmaceutical compositions of the present invention. In a non-limiting embodiment of the present invention, examples of the Fc region with a higher binding activity toward activating Fcy receptors than to inhibitory Fcy receptors (or having a selective binding activity toward inhibitory Fcy receptors) preferably include Fc regions 30 in which at least one or more amino acids selected from the group consisting of amino acids at positions 221, 222, 223, 224, 225, 227, 228, 230, 231, 232, 233, 234, 235, 236, 237, 238, 239, 240, 241, 243, 244, 245, 246, 247, 249, 250, 251, 254, 255, 256, 258, 260, 262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274, 275, 276, 278, 279, 280, 281, 282, 283, 284, 285, 286, 288, 290, 291, 292, 293, 294, 295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 311, 35 313, 315, 317, 318, 320, 322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 333, 334, 335, 336, 337, 339, 376, 377, 378, 379, 380, 382, 385, 392, 396, 421, 427, 428, 429, 434, 436, and 2023229507  12 Sep 2023 440 indicated by EU numbering mentioned above, have been altered to amino acids different from those of the native Fc region. In a non-limiting embodiment of the present invention, examples of the Fc region with a higher binding activity toward activating Fcy receptors than to inhibitory Fcy receptors (or 5 having a selective binding activity toward inhibitory Fcy receptors) preferably include Fc regions in which multiple amino acids indicated in Tables 1-1 to 1-3 have been altered to amino acids different from those of the native Fc region. Fc regions whose binding activity toward an inhibitory Fcy receptor is higher than the binding 0 activity toward an activating Fcy receptor Herein, an example of a case where the binding activity toward a certain Fcy receptor is higher than the binding activity toward another Fcy receptor is the case where the binding activity toward an inhibitory Fcy receptor is higher than the binding activity toward an activating Fcy receptor. In this case, the binding activity of the Fc region toward FcyRIIb is said to be 5 higher than the binding activity toward any of the human Fcy receptors of FcyRIa, FcyRIIa, FcyRIIIa, and / or FcyRIIIb. For example, this means that, based on an above-described analytical method, the binding activity of an antigen-binding molecule containing the Fc region toward FcyRIIb is 105% or more, preferably 110% or more, 120% or more, 130% or more, 140% or more, particularly preferably 150% or more, 160% or more, 170% or more, 180% or 0 more, 190% or more, 200% or more, 250% or more, 300% or more, 350% or more, 400% or more, 450% or more, 500% or more, 750% or more, 10-fold or more, 20-fold or more, 30-fold or more, 40-fold or more, 50-fold, 60-fold, 70-fold, 80-fold, 90-fold, or 100-fold or more as compared with the binding activity toward any of the human Fcy receptors of FcyRIa, FcyRIIa, FcyRIIIa, and / or FcyRIIIb. The Fc region with a higher binding activity toward inhibitory Fcy 25 receptors than to activating Fcy receptors may be favorably included in antigen-binding molecules of the present invention whose antigen-binding domain binds to a soluble molecule. In a non-limiting embodiment of the present invention, examples of the Fc region with a higher binding activity toward inhibitory Fcy receptors than to activating Fcy receptors (or having a selective binding activity toward inhibitory Fcy receptors) preferably include Fc regions 30 in which, of the amino acids of the above Fc region, the amino acids at 238 and 328 indicated by EU numbering are altered to amino acids different from those of the native Fc region. In a non-limiting embodiment of the present invention, examples of the Fc region with a higher binding activity toward inhibitory Fcy receptors than to activating Fcy receptors (or having a selective binding activity toward inhibitory Fcy receptors) preferably include Fc regions 35 altered at any one or more of the amino acids in the above Fc region as indicated by EU numbering: the amino acid at position 238 (indicated by EU numbering) is altered into Asp; and 2023229507  12 Sep 2023 the amino acid at position 328 (indicated by EU numbering) is altered into Glu. Furthermore, as the Fc regions having a selective binding activity toward inhibitory Fcy receptors, the Fc regions or alterations described in US 2009 / 0136485 can be suitably selected. In another non-limiting embodiment of the present invention, preferred examples 5 include Fc regions altered at any one or more of the amino acids in the above Fc region as indicated by EU numbering: the amino acid at position 238 (indicated by EU numbering) to Asp; and the amino acid at position 328 (indicated by EU numbering) to Glu. In still another non-limiting embodiment of the present invention, preferred examples include Fc regions that have one or more of the alterations exemplified in PCT / JP2012 / 054624: 0 substitution of Pro at position 238 (indicated by EU numbering) with Asp; alteration of the amino acid at position 237 (indicated by EU numbering) to Trp; alteration of the amino acid at position 237 (indicated by EU numbering) to Phe; alteration of the amino acid at position 267 (indicated by EU numbering) to Val; alteration of the amino acid at position 267 (indicated by EU numbering) to Gln; alteration of the amino acid at position 268 (indicated by EU numbering) 5 to Asn; alteration of the amino acid at position 271 (indicated by EU numbering) to Gly; alteration of the amino acid at position 326 (indicated by EU numbering) to Leu; alteration of the amino acid at position 326 (indicated by EU numbering) to Gln; alteration of the amino acid at position 326 (indicated by EU numbering) to Glu; alteration of the amino acid at position 326 (indicated by EU numbering) to Met; alteration of the amino acid at position 239 (indicated by 0   EU numbering) to Asp; alteration of the amino acid at position 267 (indicated by EU numbering) to Ala; alteration of the amino acid at position 234 (indicated by EU numbering) to Trp; alteration of the amino acid at position 234 (indicated by EU numbering) to Tyr; alteration of the amino acid at position 237 (indicated by EU numbering) to Ala; alteration of the amino acid at position 237 (indicated by EU numbering) to Asp; alteration of the amino acid at position 237 25 (indicated by EU numbering) to Glu; alteration of the amino acid at position 237 (indicated by EU numbering) to Leu; alteration of the amino acid at position 237 (indicated by EU numbering) to Met; alteration of the amino acid at position 237 (indicated by EU numbering) to Tyr; alteration of the amino acid at position 330 (indicated by EU numbering) to Lys; alteration of the amino acid at position 330 (indicated by EU numbering) to Arg, alteration of the amino acid at 30 position 233 (indicated by EU numbering) to Asp, alteration of the amino acid at position 268 (indicated by EU numbering) to Asp, alteration of the amino acid at position 268 (indicated by EU numbering) to Glu, alteration of the amino acid at position 326 (indicated by EU numbering) to Asp, alteration of the amino acid at position 326 (indicated by EU numbering) to Ser, alteration of the amino acid at position 326 (indicated by EU numbering) to Thr, alteration of the 35 amino acid at position 323 (indicated by EU numbering) to Ile, alteration of the amino acid at position 323 (indicated by EU numbering) to Leu, alteration of the amino acid at position 323 2023229507  12 Sep 2023 (indicated by EU numbering) to Met, alteration of the amino acid at position 296 (indicated by EU numbering) to Asp, alteration of the amino acid at position 326 (indicated by EU numbering) to Ala, alteration of the amino acid at position 326 (indicated by EU numbering) to Asn, and alteration of the amino acid at position 330 (indicated by EU numbering) to Met. 5 Fc regions with modified sugar chains Fc regions contained in the antigen-binding molecules provided by the present invention may include Fc regions that have been modified so that the composition of the sugar-chain-attached Fc regions has a high percentage of fucose-deficient sugar-chain-attached 0 Fc regions, or a high percentage of bisecting N-acetylglucosamine-added Fc regions. Removal of fucose residue from N-acetylglucosamine at the reducing end of N-glycoside linkage complex sugar chains bonded to the antibody Fc region is known to enhance the affinity to FcYRIIIa (Non-Patent Document 6). It is known that for IgG1 antibodies containing such Fc regions, the ADCC activity mentioned below is enhanced; therefore, antigen-binding molecules containing 5 such Fc regions are also useful as antigen-binding molecules to be contained in pharmaceutical compositions of the present invention. Examples of antibodies with fucose residue removed from N-acetylglucosamine at the reducing end of N-glycoside linkage complex sugar chains bonded to the antibody Fc regions are antibodies such as: antibodies modified by glycosylation (for example, WO 1999 / 054342); and 0 antibodies deficient in fucose attached to sugar chains (for example, WO 2000 / 061739, WO 2002 / 031140, and WO 2006 / 067913). More specifically, to produce antibodies deficient in fucose attached to sugar chains (for example, WO 2000 / 061739, WO 2002 / 031140, and WO 2006 / 067913) as another non-limiting embodiment of antibodies with fucose residue removed from N-acetylglucosamine at the 25 reducing end of N-glycoside linkage complex sugar chains bonded to the antibody Fc regions, host cells having a low ability to add fucose to sugar chains are produced by altering the activity of forming the sugar chain structure of the polypeptide to be glycosylated. Antibodies that lack fucose in their sugar chains can be collected from culture of the host cells by expressing a desired antibody gene in the host cells. Non-limiting suitable examples of the activity to form 30 the sugar chain structure of a polypeptide include the activity of a transporter or an enzyme selected from the group consisting of fucosyltransferase (EC 2.4.1.152), fucose transporter (SLC35C1), GMD (GDP-mannose-4,6-dehydratase) (EC 4.2.1.47), Fx (GDP-keto-6-deoxymannose-3,5-epimerase, 4-reductase) (EC 1.1.1.271), and GFPP (GDP-P-L-fucose pyrophosphorylase (EC 2.7.7.30). As long as these enzymes or transporters 35 can exhibit their activities, their structures are not necessarily specified. Herein, proteins that can exhibit these activities are referred to as “functional proteins”. In a non-limiting 2023229507  12 Sep 2023 embodiment, methods for altering these activities include deletion of these activities. To produce host cells deficient in these activities, known methods such as a method for destroying the genes of these functional proteins to make them unable to function may be appropriately employed (for example, WO2000 / 061739, WO2002 / 031140, and WO2006 / 067913). Host cells 5 deficient in such activities can be produced, for example, by a method that destroys the genes of these functional proteins endogenous to CHO cells, BHK cells, NS0 cells, SP2 / 0 cells, YO myeloma cells, P3X63 mouse myeloma cells, PER cells, PER.C6 cells, HEK293 cells, hybridoma cells, or such, so that the genes are unable to function. Antibodies that have a sugar chain containing bisecting GlcNAc (WO2002 / 079255, 0 etc.) are known. In a non-limiting embodiment, host cells for expressing a gene that encodes a functional protein having GnTIII (P-1,4-mannosyl-glycoprotein 4-P-N-acetylglucosaminyltransferase) (EC 2.4.1.144) activity or GalT (p-1,4-galactosyltransferase) (EC 2.4.1.38) activity are produced to prepare antibodies that have bisecting GlcNAc-containing sugar chains. In another suitable non-limiting embodiment, host 5 cells that co-express, in addition to the aforementioned functional proteins, a gene encoding a functional protein having human ManII (manosidase II) (3.2.1.114) activity, a gene encoding a functional protein having GnTI (p-1,2-acetylglucosaminyltransferase I) (EC 2.4.1.94) activity, a gene encoding a functional protein having GnTII (p-1,2-acetylglucosaminyltransferase II) (EC 2.4.1.143) activity, a gene encoding a functional protein having ManI (mannosidase) (EC 0 3.2.1.113) activity, and a-1,6-fucosyl transferase (EC 2.4.1.68), are produced (WO2004 / 065540). Antibodies with fucose residue removed from N-acetylglucosamine at the reducing end of N-glycoside linkage complex sugar chains bonded to the antibody Fc regions and antibodies having sugar chains containing bisecting GlcNAc can be produced, respectively, by transfecting 25 an expression vector containing the antibody gene into host cells with a low ability to add fucose to sugar chains, and into host cells having the activity to form bisecting GlcNAc structure-containing sugar chains. Methods for pro...

Claims

1. A method of production for an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a compound, the method comprising:(a) assaying binding of an antigen-binding domain to the antigen in the presence of a first concentration of a compound, wherein the compound is kynurenine, or a precursor or metabolite thereof;(b) assaying binding of the antigen-binding domain to the antigen in the presence of a second concentration of the compound that is different from the first concentration;(c) determining that the antigen-binding domain’s binding affinity for the antigen in the presence of the first concentration of the compound is different from the antigen-binding domain’s binding affinity for the antigen in the presence of the second concentration of the compound; and(d) selecting the antigen-binding domain based on the determination of (c), wherein either the first concentration or the second concentration, but not both, can be zero.

2. A method of production for an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a compound, the method comprising:(a) contacting (i) an antigen with (ii) a plurality of different antigen-binding domains, in the presence of a first concentration of a compound, thereby forming one or more complexes, each comprising the antigen and an antigen-binding domain, wherein the compound is kynurenine, or a precursor or metabolite thereof;(b) exposing the one or more complexes of (a) to a second concentration of the compound different from the first concentration, thereby causing at least one of the complexes to dissociate into its constituent antigen and antigen-binding domain; and(c) selecting an antigen-binding domain that dissociated from the antigen in (b), wherein either the first concentration or the second concentration, but not both, can be zero.2023229507  12 Sep 20233. A method of production for an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a compound, the method comprising:(a) contacting (i) an antigen with (ii) a plurality of different antigen-binding domains, in the presence of a first concentration of a compound, wherein the compound is kynurenine, or a precursor or metabolite thereof;(b) isolating one or more antigen-binding domains that do not bind to the antigen in the presence of the first concentration of the compound;(c) assaying binding of the one or more isolated antigen-binding domains to the antigen in the presence of a second concentration of the compound, wherein the second concentration is different than the first concentration, and wherein either the first concentration or the second concentration, but not both, can be zero; and(d) selecting an antigen-binding domain that binds to the antigen in the presence of the second concentration of the compound.

4. A method of screening for an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a compound, the method comprising:(a) assaying binding of an antigen-binding domain to the antigen in the presence of a first concentration of a compound, wherein the compound is kynurenine, or a precursor or metabolite thereof;(b) assaying binding of the antigen-binding domain to the antigen in the presence of a second concentration of the compound that is different from the first concentration;(c) determining that the antigen-binding domain’s binding affinity for the antigen in the presence of the first concentration of the compound is different from the antigen-binding domain’s binding affinity for the antigen in the presence of the second concentration of the compound; and(d) selecting the antigen-binding domain based on the determination of (c),2023229507  12 Sep 2023wherein either the first concentration or the second concentration, but not both, can be zero.

5. The method of claim 1 or 4, wherein the first concentration is higher than the second concentration, and the antigen-binding domain’s binding affinity for the antigen is higher in the first concentration than in the second concentration.

6. The method of claim 5, wherein the second concentration is zero.

7. A method of screening for an antigen-binding domain whose antigen-binding activity varies depending on the concentration of a compound, the method comprising:(a) contacting (i) an antigen with (ii) a plurality of different antigen-binding domains, in the presence of a first concentration of a compound, thereby forming one or more complexes, each comprising the antigen and an antigen-binding domain, wherein the compound is kynurenine, or a precursor or metabolite thereof;(b) exposing the one or more complexes of (a) to a second concentration of the compound different from the first concentration, thereby causing at least one of the complexes to dissociate into its constituent antigen and antigen-binding domain; and(c) selecting an antigen-binding domain that dissociated from the antigen in (b), wherein either the first concentration or the second concentration, but not both, can be zero.

8. The method of claim 2 or 7, wherein the first concentration is higher than the second concentration.

9. The method of claim 8, wherein the second concentration is zero.2023229507  12 Sep 2023activity varies depending on the concentration of a compound, the method comprising:(a) contacting (i) an antigen with (ii) a plurality of different antigen-binding domains, in the presence of a first concentration of a compound, wherein the compound is kynurenine, or a precursor or metabolite thereof;(b) isolating one or more antigen-binding domains that do not bind to the antigen in the presence of the first concentration of the compound;(c) assaying binding of the one or more isolated antigen-binding domains to the antigen in the presence of a second concentration of the compound, wherein the second concentration is different than the first concentration, and wherein either the first concentration or the second concentration, but not both, can be zero; and(d) selecting an antigen-binding domain that binds to the antigen in the presence of the second concentration of the compound.

11. The method of claim 3 or 10, wherein the first concentration is lower than the second concentration.

12. The method of claim 11, wherein the first concentration is zero.

13. The method of any one of claims 1 to 12, wherein the antigen-binding activity of the antigen-binding domain in the higher concentration of the compound compared to the antigen-binding activity of the antigen-binding domain in the lower concentration of the compound is twofold or more.

14. The method of any one of claims 1 to 12, wherein the ratio of the dissociation constant (KD) against the antigen in the lower concentration of the compound to the KD against the antigen in the higher concentration of the compound is 2 or greater.2023229507  12 Sep 202315. The method of any one of claims 1 to 14, wherein the antigen is a membrane-type molecule.

16. The method of any one of claims 1 to 15, where the antigen-binding domain has neutralizing activity.

17. The method of any one of claims 1 to 16, wherein the antigen-binding domain has cytotoxic activity.

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