Anti-TrkA antibodies and uses thereof

Abstract

Provided are antibodies or antigen-binding fragments thereof capable of specifically recognizing TrkA, and uses thereof. The antibodies comprise a CDR sequence selected from at least one of the following: heavy chain variable region CDR sequences: SEQ ID NO: 1 to SEQ ID NO: 27; light chain variable region CDR sequences: SEQ ID NO: 28 to SEQ ID NO: 54, or an amino acid sequence having at least 95% identity thereto. The antibodies according to embodiments specifically target and bind to the TrkA receptor, and can block the binding of NGF to TrkA.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to and the benefit of Chinese Patent Application No. 201910464016.0, filed with the State Intellectual Property Office of China on May 30, 2019, which is incorporated herein by reference in its entirety.

[0002] The present invention relates to the field of biotechnology. Specifically, the present invention relates to anti-TrkA antibodies and uses thereof. More specifically, the present invention relates to antibodies or antigen-binding fragments thereof, nucleic acid molecules, expression vectors, recombinant cells, pharmaceutical compositions, pharmaceutical uses, kits for detecting TrkA, and mouse B cells, capable of specifically recognizing TrkA. [Background technology]

[0003] At present, non-opioid analgesics such as non-steroidal anti-inflammatory drugs (NSAIDs) are mainly used clinically for mild to moderate pain, and opioid analgesics are mainly used for moderate to severe pain. However, NSAIDs have a "capped effect", and opioids can only effectively relieve less than 30% of non-tumor chronic pain, and 20% of patients with cancer pain have opioid tolerance. In addition, NSAIDs have potential safety risks for the gastrointestinal and circulatory systems, especially during long-term medication. With regard to opioid analgesics, years of drug improvement experiments have failed to effectively reduce their addictive properties and many other side effects, and patients are looking forward to new, safer and more effective drugs.

[0004] Nerve growth factor (NGF) is involved in the pathophysiological process of pain. Nerve growth factor (NGF) mainly activates the NGF / TrKA signaling pathway by binding to the high affinity tyrosine-nase (TrkA) receptor, affecting the release of inflammatory mediators, opening of ion channels, and promoting the growth of nerve fibers, thereby participating in the generation, conduction, and sensitization of pain. Studies have shown that blocking the NGF-TrkA signaling pathway can effectively relieve pain and hyperalgesia, and that the NGF-TrkA signaling pathway is a valid target for the development of new analgesics. With regard to NGF-TrkA targeted analgesics, the drugs currently under development are mainly divided into two categories: large molecule anti-NGF monoclonal antibodies and small molecule TrkA kinase inhibitors. Not only is the TrKA receptor the only receptor for NGF, but NGF also has a low affinity receptor P75, and the combination of NGF and P75 is important for the function of the nervous system and injury repair. As a new type of analgesic drug, large molecule anti-NGF monoclonal antibodies have great safety risks. From current clinical data, the most common toxic effects and side effects include bone tissue necrosis (possible), paresthesia, etc. However, more problems are seen when using small molecule TrkA kinase inhibitors. First, small molecule drugs generally have short half-life, frequent administration, and poor patient compliance. Second, small molecule TrkA kinase inhibitors are receptor tyrosine kinase inhibitors, and there are many receptor tyrosine kinases in the body. Therefore, it is difficult to guarantee the specificity and selectivity of drugs to specific targets, and many adverse reactions and large toxic effects and side effects are seen.

[0005] Therefore, analgesics targeting the NGF-TrkA target still need to be continuously developed and improved by scientific researchers. Summary of the Invention [Problem to be solved by the invention]

[0006] This application is based on the inventors' discovery of the following issues and facts: NGF-TrkA signaling pathway is an effective target for the development of new analgesics. If TrkA monoclonal antibody selectively targets TrkA receptor, it can not only block the activation of TrkA signaling pathway by NGF to effectively inhibit the transmission of pain signals, but also does not cause unpredictable toxic effects and side effects, such as many neurotoxicities caused by the inhibition of NGF receptor, and the use of anti-NGF antibody to over-neutralize bone and joint necrosis caused by NGF. However, since TrkA molecule is a receptor membrane protein, it is difficult to screen blocking anti-TrkA monoclonal antibody. Moreover, designing blocking TrkA receptor antibody has safety risks due to antibody-mediated immune response. Therefore, it is difficult to design and develop monoclonal antibody against TrkA. [Means for solving the problem]

[0007] However, the inventors of the present application have succeeded in screening a new type of anti-TrkA monoclonal antibody with long-acting analgesic effect. At the same time, the inventors have effectively avoided the risk of immunotoxicity by selecting specific Fc subtypes and modifying them. Specifically, in this study, a series of high-affinity anti-TrkA monoclonal antibodies that block the binding of NGF and TrkA were screened by hybridoma technology. The constant region of mouse anti-TrkA monoclonal antibody was humanized, and the variable region of mouse anti-TrkA monoclonal antibody was retained. A series of anti-TrkA chimeric monoclonal antibodies were obtained. The inventors have found that these chimeric antibody candidates obtained in the present application can not only specifically target and bind to TrkA receptor, block the binding of NGF and TrkA, and effectively inhibit pain, but also have the characteristics of low immunotoxicity and almost no antibody-dependent cell-mediated cytotoxicity (ADCC).

[0008] In a first aspect, the present invention provides an antibody or an antigen-binding fragment thereof capable of specifically recognizing TrkA. According to an embodiment of the present invention, the antibody comprises a CDR sequence selected from at least one of the following: heavy chain variable region CDR sequences: SEQ ID NO: 1 to SEQ ID NO: 27, and light chain variable region CDR sequences: SEQ ID NO: 28 to SEQ ID NO: 54, or an amino acid sequence having at least 95% identity thereto. VYSFTAYT (SEQ ID NO: 1) INPHNGGT (SEQ ID NO:2) AISRYGSSSFYFDV (SEQ ID NO: 3) GYAFTNYW (SEQ ID NO:4) FYPRTGNT (SEQ ID NO:5) ARAGTGFDY (SEQ ID NO:6) GYRFSSYW (SEQ ID NO: 7) ILPGRGII (SEQ ID NO:8) ARTDPPYFGV (SEQ ID NO: 9) GYTFSTYW (SEQ ID NO: 10) NLPGRHIT (SEQ ID NO: 11) ARGRGTYYFDY (SEQ ID NO: 12) GYSFTGYT (SEQ ID NO: 13) INPYNGGT (SEQ ID NO: 14) AFSYYGSRGFYFDY (SEQ ID NO: 15) GYSFTGYT (SEQ ID NO: 16) INPYNGGT (SEQ ID NO: 17) ASSSYRNDGNWYFDV (SEQ ID NO: 18) GYSITGYT (SEQ ID NO: 19) VNPYNGGT (SEQ ID NO: 20) AISRYGSESWYFDV (SEQ ID NO: 21) GYTFNIYW (SEQ ID NO: 22) ILPGSGNT (SEQ ID NO:23) ARTDGRGYFDY (SEQ ID NO:24) GYTFSSYW (SEQ ID NO: 25) FLPRSGKT (SEQ ID NO: 26) ARTDPPYFGV (SEQ ID NO:27) SSISY (SEQ ID NO: 28) ATS (SEQ ID NO:29) QQWSSNPPT (SEQ ID NO: 30) ENVGGY (SEQ ID NO:31) GAS (SEQ ID NO: 32) GQNYIYPFT (SEQ ID NO:33) TGAVTISNY (SEQ ID NO: 34) GTN (SEQ ID NO: 35) VLWYSNHWV (SEQ ID NO:36) QSLLHSNGITY (SEQ ID NO:37) QMS (SEQ ID NO: 38) AQNLEFPFT (SEQ ID NO:39) SSVSY (SEQ ID NO: 40) LTS (SEQ ID NO: 41) QQWSSNPPT (SEQ ID NO: 42) SSVSY (SEQ ID NO:43) DTS (SEQ ID NO: 44) QQWSSNPPT (SEQ ID NO: 45) SSVSY (SEQ ID NO:46) ATS (SEQ ID NO: 47) QQWSSNPPT (SEQ ID NO: 48) KSVSSSAYSY (SEQ ID NO: 49) LAS (SEQ ID NO:50) QHSRELPFT (SEQ ID NO:51) SNISY (SEQ ID NO:52) DTS (SEQ ID NO:53) QQWSSVPLT (SEQ ID NO:54)

[0009] The antibodies according to embodiments of the present invention can specifically target and bind to the TrkA receptor, blocking the binding of NGF and TrkA.

[0010] According to an embodiment of the invention, the antibody or antigen-binding fragment thereof may further comprise at least one of the following additional technical features: According to an embodiment of the invention, the antibody has Heavy chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, or amino acid sequences having at least 95% identity to SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, respectively; or Heavy chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6, or amino acid sequences having at least 95% identity to SEQ ID NO:4, SEQ ID NO:5 and SEQ ID NO:6, respectively; or Heavy chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9, or amino acid sequences having at least 95% identity to SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9, respectively; or A heavy chain variable region CDR1, CDR2, CDR3 sequence as shown in SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, or an amino acid sequence having at least 95% identity to SEQ ID NO: 10, SEQ ID NO: 11 and SEQ ID NO: 12, respectively; or Heavy chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 15, or amino acid sequences having at least 95% identity to SEQ ID NO: 13, SEQ ID NO: 14 and SEQ ID NO: 15, respectively; or A heavy chain variable region CDR1, CDR2, CDR3 sequence as shown in SEQ ID NO: 16, SEQ ID NO: 17 and SEQ ID NO: 18, or an amino acid sequence having at least 95% identity to SEQ ID NO: 16, SEQ ID NO: 17 and SEQ ID NO: 18, respectively; or Heavy chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO: 19, SEQ ID NO: 20 and SEQ ID NO: 21, or amino acid sequences having at least 95% identity to SEQ ID NO: 19, SEQ ID NO: 20 and SEQ ID NO: 21, respectively; or Heavy chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO:22, SEQ ID NO:23 and SEQ ID NO:24, or amino acid sequences having at least 95% identity to SEQ ID NO:22, SEQ ID NO:23 and SEQ ID NO:24, respectively; or Heavy chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO:25, SEQ ID NO:26 and SEQ ID NO:27, or amino acid sequences having at least 95% identity to SEQ ID NO:25, SEQ ID NO:26 and SEQ ID NO:27, respectively. Includes:

[0011] According to an embodiment of the invention, the antibody comprises: Light chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO:28, SEQ ID NO:29 and SEQ ID NO:30, or amino acid sequences having at least 95% identity to SEQ ID NO:28, SEQ ID NO:29 and SEQ ID NO:30, respectively; or Light chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO: 31, SEQ ID NO: 32 and SEQ ID NO: 33, or amino acid sequences having at least 95% identity to SEQ ID NO: 31, SEQ ID NO: 32 and SEQ ID NO: 33, respectively; or Light chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO: 34, SEQ ID NO: 35 and SEQ ID NO: 36, or amino acid sequences having at least 95% identity to SEQ ID NO: 34, SEQ ID NO: 35 and SEQ ID NO: 36, respectively; or Light chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO: 37, SEQ ID NO: 38 and SEQ ID NO: 39, or amino acid sequences having at least 95% identity to SEQ ID NO: 37, SEQ ID NO: 38 and SEQ ID NO: 39, respectively; or Light chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, or amino acid sequences having at least 95% identity to SEQ ID NO: 40, SEQ ID NO: 41 and SEQ ID NO: 42, respectively; or Light chain variable region CDR1, CDR2, CDR3 sequences as set forth in SEQ ID NO: 43, SEQ ID NO: 44 and SEQ ID NO: 45, or amino acid sequences having at least 95% identity to SEQ ID NO: 43, SEQ ID NO: 44 and SEQ ID NO: 45, respectively; or Light chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO: 46, SEQ ID NO: 47 and SEQ ID NO: 48, or amino acid sequences having at least 95% identity to SEQ ID NO: 46, SEQ ID NO: 47 and SEQ ID NO: 48, respectively; or Light chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO: 49, SEQ ID NO: 50 and SEQ ID NO: 51, or amino acid sequences having at least 95% identity to SEQ ID NO: 49, SEQ ID NO: 50 and SEQ ID NO: 51, respectively; or Light chain variable region CDR1, CDR2, CDR3 sequences as shown in SEQ ID NO:52, SEQ ID NO:53 and SEQ ID NO:54, or amino acid sequences having at least 95% identity to SEQ ID NO:52, SEQ ID NO:53 and SEQ ID NO:54, respectively. Includes:

[0012] According to an embodiment of the invention, the antibody or antigen-binding fragment thereof specifically recognizes the extracellular domain of TrkA.

[0013] According to embodiments of the invention, the antibody comprises at least one of a heavy chain framework region sequence and a light chain framework region sequence, and at least a portion of at least one of the heavy chain framework region sequence and the light chain framework region sequence is derived from at least one of a murine antibody, a human antibody, a primate antibody, or mutants thereof.

[0014] According to an embodiment of the present invention, the antibody has a heavy chain variable region having an amino acid sequence shown in any one of SEQ ID NO:55 to SEQ ID NO:63. EVLLQQSGPELVKPGASMKISCKASVYSFTAYTMNWVKQSHGKNLEWIGLINPHNGGTRYNQKFKGKATLTLDKSSSTAYMDLLSLTSEDSAVYYCAISRYGSSSFYFDVWGAGTTVAVSS (SEQ ID NO: 55) QVQLQQSGAELVRPGTSVKISCKASGYAFTNYWLGWMKQRPGHGLEWIGDFYPRTGNTFYNENFKGKVTLTADKSSNTAYMQLSSLTSEDSAVYLCARAGTGFDYWGQGTTLTVSS (SEQ ID NO: 56) QVQLQQSGAELMKPGASVKISCKTTGYRFSSYWIEWVKQRPGHGLEWLGEILPGRGIINYNENFRGKATFTADTSSNTAYVQLSSLTSEDSAVYFCARTDPPYFGVWGAGTTVTVSS (SEQ ID NO: 57) QVQLQQSGAELMKPGASMKISCKATGYTFSTYWIEWVKQRPGHGLEWIGENLPGRHITNYNEKFKGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARGRGTYYFDYWGQGTPLTVSS (SEQ ID NO: 58) EVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTNYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCAFSYYGSRGFYFDYWGQGTTLTVSS (SEQ ID NO: 59) EVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTRYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCASSSYRNDGNWYFDVWGAGTTVTVSS (SEQ ID NO: 60) EVQLQQSGPELVKPGASMKISCKASGYSITGYTMNWVKQSHGKNLEWIGLVNPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLKSEDSAVYYCAISRYGSESWYFDVWGAGTTVTVSS (SEQ ID NO: 61) QVHLQQSGAELMKPGASVKISCKATGYTFNIYWIDWVKQRPGHGLEWIGEILPGSGNTHYNENFKGKATMTADTSSNTAYMQLTSLTSEDSAVYYCARTDGRGYFDYWGQGTTLTVSS (SEQ ID NO: 62) QVQLQQSGAELMKPGASVKISCKATGYTFSSYWIEWVKQRPGHGLEWLGEFLPRSGKTNYNEEFRGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARTDPPYFGVWGAGTMVAVSS (SEQ ID NO: 63)

[0015] According to an embodiment of the present invention, the antibody has a light chain variable region having an amino acid sequence shown in any one of SEQ ID NO:64 to SEQ ID NO:72. QIVLSQSPAILSASPGEKVTMTCRASSSISYMHWYQQKPGSSPKPWISATSNLASGVPARFSGSGSGTSYSLTISGVEAEDAATYYCQQWSSNPPTFGGGTNLEIK (SEQ ID NO: 64) SIVMTQSPKSMSMSVGERVTLSCKASENVGGYVSWYQQKPDQSPKLLIYGASSRHTGVPDRFTGSGSETDFTLTISSVQAEDLAAYHCGQNYIYPFTFGGGTKLEIK (SEQ ID NO: 65) QAVVTQESALTTSPGETVTLTCRSSTGAVTISNYANWVQEKPDHLFTGLIGGTNNRPPGVPARFSGSLIGDKAALTITGAQTEDEAIYFCVLWYSNHWVFGGGTKLTVL (SEQ ID NO: 66) DIVMTQAAFSTPVTLGTSASISCRSSQSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLEFPFTFGSGTKLEIK (SEQ ID NO: 67) QIVLTQSPALMSASPGEKVTMTCSATSSVSYIYWYQQKPRSSPKPWIYLTSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGGGTKLEIK (SEQ ID NO: 68) DIVMTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMETEDAATYYCQQWSSNPPTFGGGTKLELK (SEQ ID NO: 69) QIVLSQSPAILSASPGEKVTMTCRATSSVSYMYWYQQKPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSSNPPTFGGGTKLEKK (SEQ ID NO: 70) DIVLTQSPASLVVSLGQRATISCRTSKSVSSSAYSYMHWYQQKPGQPPKVLIYLASNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHSRELPFTFGSGTKLEIK (SEQ ID NO: 71) QIVLTQSPPIMSASPGEKVTMTCSASSNISYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSVPLTFGAGTKLEIK (SEQ ID NO: 72)

[0016] According to embodiments of the invention, the antibody comprises at least one of a heavy chain constant region and a light chain constant region, and at least a portion of at least one of the heavy chain constant region and the light chain constant region is derived from at least one of a murine antibody, a human antibody, a primate antibody, or a mutant thereof.

[0017] According to an embodiment of the present invention, both the light chain constant region and the heavy chain constant region of the antibody are derived from a human IgG antibody or a mutant thereof. Thus, the immunogenicity of the antibody can be effectively reduced.

[0018] According to an embodiment of the invention, both the light and heavy chain constant regions of the antibody are derived from human IgG4.

[0019] According to an embodiment of the present invention, the Fc region of the antibody has S10P, F16A, L17A, R191K mutations and 229K deletion mutations compared with human IgG4 wild-type Fc, where the positioning of the above amino acid positions is based on the amino acid sequence of the human IgG4 wild-type Fc sequence shown in SEQ ID NO: 73. For example, S10P refers to the mutation of S at position 10 of the amino acid sequence shown in SEQ ID NO: 75 to P, etc. The inventors have found that after the Fc region of the antibody has the above mutations and deletions, the safety and stability of the antibody can be significantly improved, and the half-life of the antibody in the body is also significantly extended. ESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 73)

[0020] According to an embodiment of the invention, the full-length sequence of the constant region of the antibody is shown in SEQ ID NO:74 or SEQ ID NO:75. ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 74) RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 75)

[0021] Here, the full-length sequence of the constant region of the antibody shown in SEQ ID NO: 74 includes an IgG4 heavy chain constant region and an Fc region, where the sequence of the IgG4 heavy chain constant region is ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRV, and the sequence of the Fc region is ESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG. The full-length sequence of the antibody constant region shown in SEQ ID NO:75 above is an IgG4 light chain constant region.

[0022] According to an embodiment of the present invention, the antibody has a heavy chain having an amino acid sequence shown in any one of SEQ ID NOs: 76 to 84 and a light chain having an amino acid sequence shown in any one of SEQ ID NOs: 85 to 93. EVLLQQSGPELVKPGASMKISCKASVYSFTAYTMNWVKQSHGKNLEWIGLINPHNGGTRYNQKFKGKATLTLDKSSSTAYMDLLSLTSEDSAVYYCAISRYGSSSFYFDVWGA GTTVAVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 76) QIVLSQSPAILSASPGEKVTMTCRASSSISYMHWYQQKPGSSPKPWISATSSNLASGVPARFSGSGSGTSYSLTISGVEAEDAATYYCQQWSSNPPTFGGGTNLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 85) QVQLQQSGAELVRPGTSVKISCKASGYAFTNYWLGWMKQRPGHGLEWIGDFYPRTGNTFYNENFKGKVTLTADKSSNTAYMQLSSLTSEDSAVYLCARAGTGFDYWGQGTTL TVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPC PAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 77) SIVMTQSPKSMSMSVGERVTLSCKASENVGGYVSWYQQKPDQSPKLLIYGASSRHTGVPDRFTGSGSETDFTLTISSVQAEDLAAYHCGQNYIYPFTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 86) QVQLQQSGAELMKPGASVKISCKTTGYRFSSYWIEWVKQRPGHGLEWLGEILPGRGIINYNENFRGKATFTADTSSNTAYVQLSSLTSEDSAVYFCARTDPPYFGVWGAGTT VTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 78) QAVVTQESALTTSPGETVTLTCRSSTGAVTISNYANWVQEKPDHLFTGLIGGTNNRPPGVPARFSGSLIGDKAALTITGAQTEDEAIYFCVLWYSNHWVFGGGTKLTVLRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 87) QVQLQQSGAELMKPGASMKISKATGYTFSTYWIEWVKQRPGHGLEWIGENLPGRHITNYNEKFKGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARGRGTYYFDYWGQGTP LTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 79) DIVMTQAAFSTPVTLGTSASISCRSSQSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLEFPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 88) EVQLQQSGPELVKPGASMKISKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTNYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCAFSYYGSRGFYFDYWGQ GTTLTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 80) QIVLTQSPALMSASPGEKVTMTCSATSSVSYIYWYQQKPRSSPKPWIYLTSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 89) EVQLQQSGPELVKPGASMKISKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTRYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCASSSYRNDGNWYFDVWGA GTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 81) DIVMTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMETEDAATYYCQQWSSNPPTFGGGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 90) EVQLQQSGPELVKPGASMKISKASGYSITGYTMNWVKQSHGKNLEWIGLVNPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLKSEDSAVYYCAISRYGSESWYFDVWGA GTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 82) QIVLSQSPAILSASPGEKVTMTCRATSSVSYMYWYQQKPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSSNPPTFGGGTKLEKKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 91) QVHLQQSGAELMKPGASVKISCKATGYTFNIYWIDWVKQRPGHGLEWIGEILPGSGNTHYNENFKGKATMTADTSSNTAYMQLTSLTSEDSAVYYCARTDGRGYFDYWGQGTT LTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 83) DIVLTQSPASLVVSLGQRATISCRTSKSVSSSAYSYMHWYQQKPGQPPKVLIYLASNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHSRELPFTFGSGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 92) QVQLQQSGAELMKPGASVKISCKATGYTFSSYWIEWVKQRPGHGLEWLGEFLPRSGKTNYNEEFRGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARTDPPYFGVWGAGTM VAVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPP CPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 84) QIVLTQSPPIMSASPGEKVTMTCSASSNISYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSVPLTFGAGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 93)

[0023] Here, in the present application, the antibody consisting of the above SEQ ID NO: 76 and SEQ ID NO: 85 is referred to as a 20C8 chimeric antibody, the antibody consisting of the above SEQ ID NO: 77 and SEQ ID NO: 86 is referred to as a 23E12 chimeric antibody, the antibody consisting of the above SEQ ID NO: 78 and SEQ ID NO: 87 is referred to as a 27H3 chimeric antibody, the antibody consisting of the above SEQ ID NO: 79 and SEQ ID NO: 88 is referred to as a 21E5 chimeric antibody, the antibody consisting of the above SEQ ID NO: 80 and SEQ ID NO: 89 is referred to as a 2A5 chimeric antibody, the antibody consisting of the above SEQ ID NO: 81 and SEQ ID NO: 90 is referred to as a 4H4 chimeric antibody, the antibody consisting of the above SEQ ID NO: 82 and SEQ ID NO: 91 is referred to as a 22D12 chimeric antibody, the antibody consisting of the above SEQ ID NO: 83 and SEQ ID NO: 92 is referred to as a 1B9 chimeric antibody, and the antibody consisting of the above SEQ ID NO: 84 and SEQ ID NO: 93 is referred to as a 15D4 chimeric antibody.

[0024] According to embodiments of the invention, the antibody is a single chain antibody fragment, a multimeric antibody, a CDR-grafted antibody, or a small molecule antibody.

[0025] According to an embodiment of the present invention, the antibody is a single chain antibody fragment, and the antibody has the amino acid sequence shown in SEQ ID NO:94 to SEQ ID NO:111. QIVLSQSPAILSASPGEKVTMTCRASSSISYMHWYQQKPGSSPKPWISATSNLASGVPARFSGSGSGTSYSLTISGVEAEDAATYYCQQWSSNPPTFGGGTNLEIKGGGGSGGGGSGGGGSEVLLQQSGPELVKPGASMKISCKASVYSFTAYTMNWVKQSHGKNLEWIGLINPHNGGTRYNQKFKGKATLTLDKSSSTAYMDLLSLTSEDSAVYYCAISRYGSSSFYFDVWGAGTTVAVSS (SEQ ID NO: 94) EVLLQQSGPELVKPGASMKISCKASVYSFTAYTMNWVKQSHGKNLEWIGLINPHNGGTRYNQKFKGKATLTLDKSSSTAYMDLLSLTSEDSAVYYCAISRYGSSSFYFDVWGAGTTVAVSSGGGGSGGGGSGGGGSQIVLSQSPAILSASPGEKVTMTCRASSSISYMHWYQQKPGSSPKPWISATSNLASGVPARFSGSGSGTSYSLTISGVEAEDAATYYCQQWSSNPPTFGGGTNLEIK (SEQ ID NO: 95) SIVMTQSPKSMSMSVGERVTLSCKASENVGGYVSWYQQKPDQSPKLLIYGASSRHTGVPDRFTGSGSETDFTLTISSVQAEDLAAYHCGQNYIYPFTFGGGTKLEIKGGGGSGGGGSGGGGSQVQLQQSGAELVRPGTSVKISCKASGYAFTNYWLGWMKQRPGHGLEWIGDFYPRTGNTFYNENFKGKVTLTADKSSNTAYMQLSSLTSEDSAVYLCARAGTGFDYWGQGTTLTVSS (SEQ ID NO: 96) QVQLQQSGAELVRPGTSVKISCKASGYAFTNYWLGWMKQRPGHGLEWIGDFYPRTGNTFYNENFKGKVTLTADKSSNTAYMQLSSLTSEDSAVYLCARAGTGFDYWGQGTTLTVSSGGGGSGGGGSGGGGSSIVMTQSPKSMSMSVGERVTLSCKASENVGGYVSWYQQKPDQSPKLLIYGASSRHTGVPDRFTGSGSETDFTLTISSVQAEDLAAYHCGQNYIYPFTFGGGTKLEIK (SEQ ID NO: 97) QAVVTQESALTTSPGETVTLTCRSSTGAVTISNYANWVQEKPDHLFTGLIGGTNNRPPGVPARFSGSLIGDKAALTITGAQTEDEAIYFCVLWYSNHWVFGGGTKLTVLGGGGSGGGGSGGGGSQVQLQQSGAELMKPGASVKISCKTTGYRFSSYWIEWVKQRPGHGLEWLGEILPGRGIINYNENFRGKATFTADTSSNTAYVQLSSLTSEDSAVYFCARTDPPYFGVWGAGTTVTVSS (SEQ ID NO: 98) QVQLQQSGAELMKPGASVKISCKTTGYRFSSYWIEWVKQRPGHGLEWLGEILPGRGIINYNENFRGKATFTADTSSNTAYVQLSSLTSEDSAVYFCARTDPPYFGVWGAGTTVTVSSGGGGSGGGGSGGGGSQAVVTQESALTTSPGETVTLTCRSSTGAVTISNYANWVQEKPDHLFTGLIGGTNNRPPGVPARFSGSLIGDKAALTITGAQTEDEAIYFCVLWYSNHWVFGGGTKLTVL (SEQ ID NO: 99) DIVMTQAAFSTPVTLGTSASISCRSSQSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLEFPFTFGSGTKLEIKGGGGSGGGGSGGGGSQVQLQQSGAELMKPGASMKISCKATGYTFSTYWIEWVKQRPGHGLEWIGENLPGRHITNYNEKFKGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARGRGTYYFDYWGQGTPLTVSS (sequence number 100) QVQLQQSGAELMKPGASMKISCKATGYTFSTYWIEWVKQRPGHGLEWIGENLPGRHITNYNEKFKGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARGRGTYYFDYWGQGTPLTVSSGGGGSGGGGSGGGGSDIVMTQAAFSTPVTLGTSASISCRSSQSLLHSNGITYLYWYLQKPGQSPQLLIYQMSNLASGVPDRFSSSGSGTDFTLRISRVEAEDVGVYYCAQNLEFPFTFGSGTKLEIK (SEQ ID NO: 101) QIVLTQSPALMSASPGEKVTMTCSATSSVSYIYWYQQKPRSSPKPWIYLTSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGGGTKLEIKGGGGSGGGGSGGGGSEVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTNYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCAFSYYGSRGFYFDYWGQGTTLTVSS (SEQ ID NO: 102) EVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTNYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCAFSYYGSRGFYFDYWGQGTTLTVSSGGGGSGGGGSGGGGSQIVLTQSPALMSASPGEKVTMTCSATSSVSYIYWYQQKPRSSPKPWIYLTSNLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPPTFGGGTKLEIK (SEQ ID NO: 103) DIVMTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMETEDAATYYCQQWSSNPPTFGGGTKLELKGGGGSGGGGSGGGGSEVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTRYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCASSSYRNDGNWYFDVWGAGTTVTVSS (SEQ ID NO: 104) EVQLQQSGPELVKPGASMKISCKASGYSFTGYTMNWVKQSHGKNLEWIGLINPYNGGTRYNQKFKGKATLTVDKSSSTAYMELLSLTSEDSAVYYCASSSYRNDGNWYFDVWGAGTTVTVSSGGGGSGGGGSGGGGSDIVMTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMETEDAATYYCQQWSSNPPTFGGGTKLELK (SEQ ID NO: 105) QIVLSQSPAILSASPGEKVTMTCRATSSVSYMYWYQQKPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSSNPPTFGGGTKLEKKGGGGSGGGGSGGGGSEVQLQQSGPELVKPGASMKISCKASGYSITGYTMNWVKQSHGKNLEWIGLVNPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLKSEDSAVYYCAISRYGSESWYFDVWGAGTTVTVSS (SEQ ID NO: 106) EVQLQQSGPELVKPGASMKISCKASGYSITGYTMNWVKQSHGKNLEWIGLVNPYNGGTSYNQKFKGKATLTVDKSSSTAYMELLSLKSEDSAVYYCAISRYGSESWYFDVWGAGTTVTVSSGGGGSGGGGSGGGGSQIVLSQSPAILSASPGEKVTMTCRATSSVSYMYWYQQKPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSSNPPTFGGGTKLEKK (SEQ ID NO: 107) DIVLTQSPASLVVSLGQRATISCRTSKSVSSSAYSYMHWYQQKPGQPPKVLIYLASNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHSRELPFTFGSGTKLEIKGGGGSGGGGSGGGGSQVHLQQSGAELMKPGASVKISCKATGYTFNIYWIDWVKQRPGHGLEWIGEILPGSGNTHYNENFKGKATMTADTSSNTAYMQLTSLTSEDSAVYYCARTDGRGYFDYWGQGTTLTVSS (SEQ ID NO: 108) QVHLQQSGAELMKPGASVKISCKATGYTFNIYWIDWVKQRPGHGLEWIGEILPGSGNTHYNENFKGKATMTADTSSNTAYMQLTSLTSEDSAVYYCARTDGRGYFDYWGQGTTLTVSSGGGGSGGGGSGGGGSDIVLTQSPASLVVSLGQRATISCRTSKSVSSSAYSYMHWYQQKPGQPPKVLIYLASNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHSRELPFTFGSGTKLEIK (SEQ ID NO: 109) QIVLTQSPPIMSASPGEKVTMTCSASSNISYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSVPLTFGAGTKLEIKGGGGSGGGGSGGGGSQVQLQQSGAELMKPGASVKISCKATGYTFSSYWIEWVKQRPGHGLEWLGEFLPRSGKTNYNEEFRGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARTDPPYFGVWGAGTMVAVSS (SEQ ID NO: 110) QVQLQQSGAELMKPGASVKISCKATGYTFSSYWIEWVKQRPGHGLEWLGEFLPRSGKTNYNEEFRGKATFTADTSSNTAYMQLSSLTSEDSAVYYCARTDPPYFGVWGAGTMVAVSSGGGGSGGGGSGGGGSQIVLTQSPPIMSASPGEKVTMTCSASSNISYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSVPLTFGAGTKLEIK (SEQ ID NO: 111)

[0026] In the present application, the antibody having the amino acid sequence shown in SEQ ID NO: 94 or SEQ ID NO: 95 is referred to as a 20C8 single chain antibody fragment, the antibody having the amino acid sequence shown in SEQ ID NO: 96 or SEQ ID NO: 97 is referred to as a 23E12 single chain antibody fragment, the antibody having the amino acid sequence shown in SEQ ID NO: 98 or SEQ ID NO: 99 is referred to as a 27H3 single chain antibody fragment, the antibody having the amino acid sequence shown in SEQ ID NO: 100 or SEQ ID NO: 101 is referred to as a 21E5 single chain antibody fragment, and the antibody having the amino acid sequence shown in SEQ ID NO: 102 or SEQ ID NO: 103 is referred to as a 21E5 single chain antibody fragment. An antibody having the amino acid sequence shown in SEQ ID NO:104 or SEQ ID NO:105 is referred to as a 4H4 single-chain antibody fragment, an antibody having the amino acid sequence shown in SEQ ID NO:106 or SEQ ID NO:107 is referred to as a 22D12 single-chain antibody fragment, an antibody having the amino acid sequence shown in SEQ ID NO:108 or SEQ ID NO:109 is referred to as a 1B9 single-chain antibody fragment, and an antibody having the amino acid sequence shown in SEQ ID NO:110 or SEQ ID NO:111 is referred to as a 15D4 single-chain antibody fragment. Among them, antibodies having amino acid sequences shown in SEQ ID NO: 94, SEQ ID NO: 96, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 102, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 108, and SEQ ID NO: 110 can be represented as VL-Link-VH (VL represents the light chain variable region, VH represents the heavy chain variable region, and Link represents the connecting chain that connects VL and VH) from the N-terminus to the C-terminus, and antibodies having amino acid sequences shown in SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 99, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105, SEQ ID NO: 107, SEQ ID NO: 109, and SEQ ID NO: 111 can be represented as VH-Link-VL (VL represents the light chain variable region, VH represents the heavy chain variable region, and Link represents the connecting chain that connects VL and VH) from the N-terminus to the C-terminus.

[0027] According to embodiments of the present invention, the small molecule antibody includes at least one of a Fab antibody, an Fv antibody, a single domain antibody, and a minimal recognition unit.

[0028] In a second aspect of the present invention, the present invention provides a nucleic acid molecule. According to an embodiment of the present invention, the nucleic acid molecule encodes the above-mentioned antibody or antigen-binding fragment thereof. The antibody or antigen-binding fragment encoded by the nucleic acid molecule according to an embodiment of the present invention can specifically target and bind to the TrkA receptor and block the binding of NGF and TrkA, and has the advantages of low immunogenicity and low ADCC.

[0029] According to an embodiment of the present invention, the nucleic acid molecule may further comprise at least one of the following additional technical features: According to an embodiment of the present invention, the nucleic acid molecule is DNA.

[0030] According to an embodiment of the present invention, the nucleic acid molecule has a nucleotide sequence as set forth in any one of SEQ ID NOs: 112 to 120, or a nucleotide sequence as set forth in any one of SEQ ID NOs: 121 to 129, or a nucleotide sequence as set forth in any one of SEQ ID NOs: 130 to 147. CAGATCGTGCTGAGCCAGAGCCCAGCTATTCTGTCCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTAGGGCCAGCTCCAGCATCAGCTACATGCACTGGTACCAGCAGAAGCCCGGCTCCTCCCCAAAGCCTTGGATCAGCGCCACTAGCAATCTGGCCAGCGGCGTGCCAGCCAGATTCAGCGGAAGCGGCAGCGGCACTAGCTACTCTCTGACTATCTCCGGCGTGGAAGCTGAGGATGCCGCCACTTACTACTGCCAGCAGTGGTCCAGCAACCCTCCTACTTTCGGCGGCGGCACAAATCTGGAGATCAAGCGGACCGTGGCTGCCCCCTCCGTGTTCATCTTCCCCCCTTCCGACGAGCAGCTGAAGTCCGGCACCGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC (SEQ ID NO: 121) AGCATCGTCATGACACAAAGCCCTAAGAGCATGAGCATGAGCGTGGGCGAGAGAGTGACTCTGAGCTGTAAGGCCAGCGAGAATGTGGGCGGCTACGTGAGCTGGTATCAGCAGAAGCCAGATCAGAGCCCAAAGCTGCTGATCTACGGCGCCAGCAGCAGACACACTGGCGTGCCAGATAGGTTCACTGGCAGCGGCTCCGAGACAGACTTCACTCTGACTATCAGCAGCGTCCAAGCCGAAGATCTGGCCGCCTATCACTGCGGCCAGAACTACATCTACCCATTCACATTCGGCGGCGGCACAAAGCTGGAGATCAAGCGGACCGTGGCTGCCCCCTCCGTGTTCATCTTCCCCCCTTCCGACGAGCAGCTGAAGTCCGGCACCGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO: 122) (query number 123) GATATCGTCATGACTCAAGCCGCCTTCAGCACTCCAGTCACTCTCGGCACAAGCGCCAGCATCAGCTGTAGGTCCAGCCAGTCTCTGCTGCACAGCAACGGCATCACTTATCTGTACTGGTATCTGCAGAAGCCCGGCCAAAGCCCACAGCTGCTGATCTACCAGATGAGCAATCTGGCCAGCGGCGTGCCAGATAGATTCAGCAGCAGCGGAAGCGGAACAGACTTCACACTGAGGATCTCTAGGGTGGAAGCCGAAGATGTGGGCGTCTATTACTGCGCCCAGAACCTCGAGTTCCCTTTCACATTCGGCAGCGGCACTAAGCTGGAGATCAAGCGGACCGTGGCTGCCCCCTCCGTGTTCATCTTCCCCCCTTCCGACGAGCAGCTGAAGTCCGGCACCGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO: 124) CAGATCGTGCTGACACAGAGCCCAGCTCTGATGAGCGCCAGCCCCGGCGAGAAGGTCACAATGACTTGCAGCGCCACATCCAGCGTGAGCTACATCTACTGGTACCAGCAGAAGCCTAGGAGCAGCCCTAAGCCTTGGATCTACCTCACAAGCAATCTGGCCAGCGGAGTGCCAGCTAGGTTCAGCGGAAGCGGCAGCGGCACAAGCTACTCTCTGACAATCTCCAGCATGGAAGCCGAAGATGCCGCCACTTACTACTGCCAGCAGTGGAGCAGCAATCCACCTACATTCGGAGGCGGCACTAAGCTGGAGATCAAGCGGACCGTGGCTGCCCCCTCCGTGTTCATCTTCCCCCCTTCCGACGAGCAGCTGAAGTCCGGCACCGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO: 125) GATATCGTGATGACTCAGTCCCCAGCCATCATGTCCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGCTCCGCCAGCAGCAGCGTGAGCTACATGCACTGGTACCAGCAGAAGAGCGGCACATCCCCAAAGAGGTGGATCTACGACACAAGCAAGCTGGCCAGCGGCGTGCCAGCCAGATTCAGCGGCTCCGGCAGCGGAACAAGCTACTCTCTGACTATCAGCAGCATGGAGACAGAGGACGCTGCCACTTACTACTGCCAGCAGTGGAGCAGCAATCCTCCAACTTTCGGCGGAGGCACTAAGCTGGAGCTGAAGCGGACCGTGGCTGCCCCCTCCGTGTTCATCTTCCCCCCTTCCGACGAGCAGCTGAAGTCCGGCACCGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC (SEQ ID NO: 126) CAGATTGTGCTGTCCCAGTCCCCAGCCATTCTGAGCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTAGGGCCACAAGCAGCGTGAGCTACATGTACTGGTACCAGCAGAAGCCCGGCAGCAGCCCTAAGCCTTGGATCTACGCCACAAGCAATCTGGCCAGCGGCGTCCCAGCTAGATTTAGCGGCAGCGGATCCGGCACTAGCTATTCTCTGACTATCTCTAGGGTCGAGGCCGAAGATGCCGCCACATACTACTGCCAGCAGTGGTCCTCCAATCCTCCAACATTCGGCGGCGGAACTAAGCTGGAGAAGAAGAGGACAGTGGCTGCCCCTTCCGTGTTCATCTTCCCTCCAAGCGACGAGCAGCTGAAGTCCGGCACTGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO: 127) GATATCGTGCTGACTCAGAGCCCAGCCTCTCTGGTGGTGTCTCTGGGACAGAGGGCCACAATCAGCTGTAGGACTTCCAAGAGCGTGAGCAGCTCCGCCTACAGCTACATGCACTGGTACCAGCAGAAGCCCGGCCAGCCTCCTAAGGTGCTGATCTATCTGGCCAGCAATCTGGAGAGCGGCGTCCCAGCTAGATTCAGCGGCTCCGGAAGCGGCACTGACTTCACTCTGAACATCCACCCAGTGGAAGAGGAGGATGCCGCCACATACTACTGCCAGCACTCTAGGGAGCTGCCTTTCACATTTGGCAGCGGAACTAAGCTGGAGATCAAGAGGACTGTCGCCGCCCCTAGCGTGTTCATCTTCCCTCCAAGCGATGAGCAGCTGAAGAGCGGCACTGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO: 128) CAAATTGTTCTCACACAGAGCCCTCCTATCATGAGCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTTCCGCCAGCAGCAACATCAGCTACATGCACTGGTACCAGCAGAAGTCCGGCACAAGCCCAAAGAGGTGGATCTACGACACAAGCAAGCTGGCCAGCGGCGTGCCAGCCAGATTTAGCGGCTCCGGAAGCGGCACTAGCTACTCTCTGACAATCAGCAGCATGGAAGCCGAGGACGCCGCTACATACTACTGCCAGCAGTGGAGCTCCGTCCCACTGACTTTCGGCGCTGGCACTAAGCTGGAGATCAAGAGGACTGTGGCCGCCCCTTCCGTGTTCATCTTCCCTCCTAGCGACGAACAGCTCAAGAGCGGCACTGCTAGCGTGGTGTGTTTACTGAACAACTTCTACCCTCGTGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCTTTACAGTCCGGCAACTCCCAAGAATCCGTGACCGAGCAAGATTCCAAGGACTCCACCTACTCTTTATCCTCCACTTTAACTTTATCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCTTGTGAGGTGACCCATCAAGGTTTATCCTCCCCCGTGACCAAGTCCTTCAATCGTGGCGAGTGCTGATGAGAATTC(SEQ ID NO: 129) CAGATCGTGCTGAGCCAGAGCCCAGCTATTCTGTCCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTAGGGCCAGCTCCAGCATCAGCTACATGCACTGGTACCAGCAGAAGCCCGGCTCCTCCCCAAAGCCTTGGATCAGCGCCACTAGCAATCTGGCCAGCGGCGTGCCAGCCAGATTCAGCGGAAGCGGCAGCGGCACTAGCTACTCTCTGACTATCTCCGGCGTGGAAGCTGAGGATGCCGCCACTTACTACTGCCAGCAGTGGTCCAGCAACCCTCCTACTTTCGGCGGCGGCACAAATCTGGAGATCAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGAAGTGCTGCTGCAGCAGAGCGGACCAGAGCTGGTCAAGCCCGGCGCCTCCATGAAGATCAGCTGTAAGGCCTCCGTGTACAGCTTCACAGCCTACACAATGAACTGGGTGAAGCAGAGCCACGGCAAGAATCTGGAGTGGATCGGACTGATCAACCCACACAATGGCGGCACTAGGTACAACCAGAAGTTCAAGGGCAAGGCCACACTGACTCTGGATAAGTCCAGCAGCACTGCCTACATGGATCTGCTGTCTCTGACAAGCGAGGACAGCGCCGTCTACTATTGCGCCATCTCTAGGTACGGCAGCAGCAGCTTCTACTTCGATGTGTGGGGCGCCGGCACAACAGTGGCTGTGAGCAGC (SEQ ID NO: 130) GAAGTGCTGCTGCAGCAGAGCGGACCAGAGCTGGTCAAGCCCGGCGCCTCCATGAAGATCAGCTGTAAGGCCTCCGTGTACAGCTTCACAGCCTACACAATGAACTGGGTGAAGCAGAGCCACGGCAAGAATCTGGAGTGGATCGGACTGATCAACCCACACAATGGCGGCACTAGGTACAACCAGAAGTTCAAGGGCAAGGCCACACTGACTCTGGATAAGTCCAGCAGCACTGCCTACATGGATCTGCTGTCTCTGACAAGCGAGGACAGCGCCGTCTACTATTGCGCCATCTCTAGGTACGGCAGCAGCAGCTTCTACTTCGATGTGTGGGGCGCCGGCACAACAGTGGCTGTGAGCAGCGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAGATCGTGCTGAGCCAGAGCCCAGCTATTCTGTCCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTAGGGCCAGCTCCAGCATCAGCTACATGCACTGGTACCAGCAGAAGCCCGGCTCCTCCCCAAAGCCTTGGATCAGCGCCACTAGCAATCTGGCCAGCGGCGTGCCAGCCAGATTCAGCGGAAGCGGCAGCGGCACTAGCTACTCTCTGACTATCTCCGGCGTGGAAGCTGAGGATGCCGCCACTTACTACTGCCAGCAGTGGTCCAGCAACCCTCCTACTTTCGGCGGCGGCACAAATCTGGAGATCAAG (SEQ ID NO: 131) AGCATCGTCATGACACAAAGCCCTAAGAGCATGAGCATGAGCGTGGGCGAGAGAGTGACTCTGAGCTGTAAGGCCAGCGAGAATGTGGGCGGCTACGTGAGCTGGTATCAGCAGAAGCCAGATCAGAGCCCAAAGCTGCTGATCTACGGCGCCAGCAGCAGACACACTGGCGTGCCAGATAGGTTCACTGGCAGCGGCTCCGAGACAGACTTCACTCTGACTATCAGCAGCGTCCAAGCCGAAGATCTGGCCGCCTATCACTGCGGCCAGAACTACATCTACCCATTCACATTCGGCGGCGGCACAAAGCTGGAGATCAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAAGTGCAGCTCCAGCAGAGCGGAGCTGAGCTGGTGAGACCCGGCACTAGCGTGAAGATCAGCTGTAAGGCCAGCGGCTACGCCTTCACTAATTACTGGCTGGGCTGGATGAAGCAGAGACCCGGCCATGGACTGGAGTGGATCGGCGACTTCTACCCAAGGACTGGCAACACTTTCTACAACGAGAACTTTAAGGGCAAGGTGACTCTGACTGCCGATAAGTCCAGCAACACTGCCTACATGCAGCTGTCCTCTCTGACTAGCGAAGATAGCGCCGTGTATCTGTGTGCTAGGGCTGGCACTGGCTTCGATTACTGGGGCCAAGGCACAACACTGACAGTGAGCAGC (SEQ ID NO: 132) (query number 133) (query number 134) (query number 135) (query number 136) CAAGTGCAGCTGCAGCAGAGCGGAGCCGAGCTGATGAAGCCCGGCGCCTCCATGAAGATCAGCTGCAAGGCCACTGGCTACACATTCAGCACATACTGGATCGAGTGGGTCAAGCAGAGACCCGGCCACGGACTGGAGTGGATCGGAGAGAATCTGCCCGGAAGGCACATCACTAACTACAACGAGAAGTTCAAGGGCAAGGCCACTTTCACAGCCGACACTAGCAGCAACACTGCCTACATGCAGCTCAGCTCTCTGACAAGCGAAGATAGCGCCGTGTACTACTGTGCTAGGGGAAGGGGCACTTACTACTTCGATTACTGGGGCCAAGGCACTCCACTGACTGTGTCCAGCGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGATATCGTCATGACTCAAGCCGCCTTCAGCACTCCAGTCACTCTCGGCACAAGCGCCAGCATCAGCTGTAGGTCCAGCCAGTCTCTGCTGCACAGCAACGGCATCACTTATCTGTACTGGTATCTGCAGAAGCCCGGCCAAAGCCCACAGCTGCTGATCTACCAGATGAGCAATCTGGCCAGCGGCGTGCCAGATAGATTCAGCAGCAGCGGAAGCGGAACAGACTTCACACTGAGGATCTCTAGGGTGGAAGCCGAAGATGTGGGCGTCTATTACTGCGCCCAGAACCTCGAGTTCCCTTTCACATTCGGCAGCGGCACTAAGCTGGAGATCAAG(SEQ ID NO: 137) CAGATCGTGCTGACACAGAGCCCAGCTCTGATGAGCGCCAGCCCCGGCGAGAAGGTCACAATGACTTGCAGCGCCACATCCAGCGTGAGCTACATCTACTGGTACCAGCAGAAGCCTAGGAGCAGCCCTAAGCCTTGGATCTACCTCACAAGCAATCTGGCCAGCGGAGTGCCAGCTAGGTTCAGCGGAAGCGGCAGCGGCACAAGCTACTCTCTGACAATCTCCAGCATGGAAGCCGAAGATGCCGCCACTTACTACTGCCAGCAGTGGAGCAGCAATCCACCTACATTCGGAGGCGGCACTAAGCTGGAGATCAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGAGGTGCAGCTCCAACAGAGCGGACCAGAGCTGGTGAAACCCGGCGCCAGCATGAAGATCAGCTGTAAGGCCTCCGGCTACAGCTTCACTGGCTACACTATGAACTGGGTGAAGCAGTCCCACGGCAAGAATCTGGAGTGGATCGGACTGATCAACCCATACAACGGCGGCACAAACTACAACCAGAAGTTCAAGGGCAAGGCCACTCTGACAGTCGATAAGAGCTCCAGCACTGCCTACATGGAGCTGCTGAGCCTCACTAGCGAGGACAGCGCTGTGTACTACTGTGCCTTCTCCTACTACGGCTCTAGGGGCTTCTACTTCGATTACTGGGGCCAAGGCACAACACTGACAGTGTCCAGC(SEQ ID NO: 138) GAGGTGCAGCTCCAACAGAGCGGACCAGAGCTGGTGAAACCCGGCGCCAGCATGAAGATCAGCTGTAAGGCCTCCGGCTACAGCTTCACTGGCTACACTATGAACTGGGTGAAGCAGTCCCACGGCAAGAATCTGGAGTGGATCGGACTGATCAACCCATACAACGGCGGCACAAACTACAACCAGAAGTTCAAGGGCAAGGCCACTCTGACAGTCGATAAGAGCTCCAGCACTGCCTACATGGAGCTGCTGAGCCTCACTAGCGAGGACAGCGCTGTGTACTACTGTGCCTTCTCCTACTACGGCTCTAGGGGCTTCTACTTCGATTACTGGGGCCAAGGCACAACACTGACAGTGTCCAGCGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAGATCGTGCTGACACAGAGCCCAGCTCTGATGAGCGCCAGCCCCGGCGAGAAGGTCACAATGACTTGCAGCGCCACATCCAGCGTGAGCTACATCTACTGGTACCAGCAGAAGCCTAGGAGCAGCCCTAAGCCTTGGATCTACCTCACAAGCAATCTGGCCAGCGGAGTGCCAGCTAGGTTCAGCGGAAGCGGCAGCGGCACAAGCTACTCTCTGACAATCTCCAGCATGGAAGCCGAAGATGCCGCCACTTACTACTGCCAGCAGTGGAGCAGCAATCCACCTACATTCGGAGGCGGCACTAAGCTGGAGATCAAG(SEQ ID NO: 139) GATATCGTGATGACTCAGTCCCCAGCCATCATGTCCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGCTCCGCCAGCAGCAGCGTGAGCTACATGCACTGGTACCAGCAGAAGAGCGGCACATCCCCAAAGAGGTGGATCTACGACACAAGCAAGCTGGCCAGCGGCGTGCCAGCCAGATTCAGCGGCTCCGGCAGCGGAACAAGCTACTCTCTGACTATCAGCAGCATGGAGACAGAGGACGCTGCCACTTACTACTGCCAGCAGTGGAGCAGCAATCCTCCAACTTTCGGCGGAGGCACTAAGCTGGAGCTGAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGAAGTGCAGCTGCAACAGAGCGGCCCAGAGCTCGTGAAGCCCGGCGCCAGCATGAAGATCAGCTGCAAGGCCAGCGGCTACAGCTTCACTGGCTACACAATGAACTGGGTCAAGCAGAGCCACGGAAAAAATCTGGAGTGGATCGGACTGATCAACCCTTACAACGGCGGCACAAGGTACAATCAGAAGTTCAAGGGCAAGGCCACTCTCACTGTGGATAAGAGCAGCAGCACTGCCTACATGGAGCTGCTGTCTCTGACAAGCGAGGATAGCGCCGTGTACTACTGCGCCAGCTCCTCCTATAGGAACGACGGCAACTGGTACTTCGATGTGTGGGGCGCCGGCACTACTGTGACAGTGAGCTCC(SEQ ID NO: 140) (query number 141) CAGATTGTGCTGTCCCAGTCCCCAGCCATTCTGAGCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTAGGGCCACAAGCAGCGTGAGCTACATGTACTGGTACCAGCAGAAGCCCGGCAGCAGCCCTAAGCCTTGGATCTACGCCACAAGCAATCTGGCCAGCGGCGTCCCAGCTAGATTTAGCGGCAGCGGATCCGGCACTAGCTATTCTCTGACTATCTCTAGGGTCGAGGCCGAAGATGCCGCCACATACTACTGCCAGCAGTGGTCCTCCAATCCTCCAACATTCGGCGGCGGAACTAAGCTGGAGAAGAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTGAAGTGCAGCTGCAGCAGAGCGGACCAGAGCTGGTCAAGCCCGGCGCCAGCATGAAGATCAGCTGTAAGGCCAGCGGCTACAGCATCACTGGCTACACTATGAACTGGGTGAAGCAGAGCCACGGCAAGAACCTCGAGTGGATTGGCCTCGTGAACCCATACAACGGCGGCACTTCCTACAACCAGAAGTTCAAAGGCAAGGCCACACTCACAGTCGATAAGTCCAGCTCCACAGCCTACATGGAGCTGCTGTCTCTGAAGAGCGAGGATAGCGCTGTCTACTACTGTGCCATCAGCAGATACGGCAGCGAGAGCTGGTACTTCGACGTGTGGGGCGCCGGCACAACAGTGACAGTGAGCAGC (SEQ ID NO: 142) (query number 143) GATATCGTGCTGACTCAGAGCCCAGCCTCTCTGGTGGTGTCTCTGGGACAGAGGGCCACAATCAGCTGTAGGACTTCCAAGAGCGTGAGCAGCTCCGCCTACAGCTACATGCACTGGTACCAGCAGAAGCCCGGCCAGCCTCCTAAGGTGCTGATCTATCTGGCCAGCAATCTGGAGAGCGGCGTCCCAGCTAGATTCAGCGGCTCCGGAAGCGGCACTGACTTCACTCTGAACATCCACCCAGTGGAAGAGGAGGATGCCGCCACATACTACTGCCAGCACTCTAGGGAGCTGCCTTTCACATTTGGCAGCGGAACTAAGCTGGAGATCAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAAGTGCATCTGCAGCAGAGCGGCGCTGAGCTGATGAAGCCCGGCGCCAGCGTGAAGATTAGCTGCAAGGCCACTGGCTACACATTCAACATCTACTGGATCGACTGGGTGAAGCAGAGGCCCGGCCACGGACTGGAATGGATCGGCGAAATTCTGCCCGGCAGCGGCAACACTCACTACAACGAGAACTTCAAGGGCAAGGCCACAATGACAGCCGACACAAGCTCCAACACTGCTTACATGCAGCTGACTTCTCTGACTAGCGAGGACAGCGCCGTGTACTATTGCGCTAGGACAGACGGAAGGGGCTACTTCGATTACTGGGGCCAAGGCACTACACTCACAGTGAGCAGC(SEQ ID NO: 144) (query number 145) CAAATTGTTCTCACACAGAGCCCTCCTATCATGAGCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTTCCGCCAGCAGCAACATCAGCTACATGCACTGGTACCAGCAGAAGTCCGGCACAAGCCCAAAGAGGTGGATCTACGACACAAGCAAGCTGGCCAGCGGCGTGCCAGCCAGATTTAGCGGCTCCGGAAGCGGCACTAGCTACTCTCTGACAATCAGCAGCATGGAAGCCGAGGACGCCGCTACATACTACTGCCAGCAGTGGAGCTCCGTCCCACTGACTTTCGGCGCTGGCACTAAGCTGGAGATCAAGGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAGGTTCAGCTGCAGCAGTCCGGCGCTGAGCTCATGAAGCCCGGCGCCAGCGTGAAGATCAGCTGCAAGGCCACTGGCTACACATTCAGCAGCTACTGGATCGAGTGGGTGAAGCAGAGACCCGGCCACGGACTGGAATGGCTGGGAGAGTTTCTGCCTAGAAGCGGCAAGACAAACTACAACGAGGAGTTTAGGGGCAAGGCTACATTCACTGCCGACACATCCAGCAACACTGCCTACATGCAGCTGAGCAGCCTCACAAGCGAGGATTCCGCCGTCTACTACTGTGCTAGGACTGATCCTCCTTACTTCGGAGTGTGGGGCGCTGGCACAATGGTGGCTGTGAGCAGC (SEQ ID NO: 146) CAGGTTCAGCTGCAGCAGTCCGGCGCTGAGCTCATGAAGCCCGGCGCCAGCGTGAAGATCAGCTGCAAGGCCACTGGCTACACATTCAGCAGCTACTGGATCGAGTGGGTGAAGCAGAGACCCGGCCACGGACTGGAATGGCTGGGAGAGTTTCTGCCTAGAAGCGGCAAGACAAACTACAACGAGGAGTTTAGGGGCAAGGCTACATTCACTGCCGACACATCCAGCAACACTGCCTACATGCAGCTGAGCAGCCTCACAAGCGAGGATTCCGCCGTCTACTACTGTGCTAGGACTGATCCTCCTTACTTCGGAGTGTGGGGCGCTGGCACAATGGTGGCTGTGAGCAGCGGTGGCGGTGGCTCGGGCGGTGGTGGGTCGGGTGGCGGCGGATCTCAAATTGTTCTCACACAGAGCCCTCCTATCATGAGCGCCAGCCCCGGCGAGAAGGTGACTATGACTTGTTCCGCCAGCAGCAACATCAGCTACATGCACTGGTACCAGCAGAAGTCCGGCACAAGCCCAAAGAGGTGGATCTACGACACAAGCAAGCTGGCCAGCGGCGTGCCAGCCAGATTTAGCGGCTCCGGAAGCGGCACTAGCTACTCTCTGACAATCAGCAGCATGGAAGCCGAGGACGCCGCTACATACTACTGCCAGCAGTGGAGCTCCGTCCCACTGACTTTCGGCGCTGGCACTAAGCTGGAGATCAAG (SEQ ID NO: 147)

[0031] Here, in the present application, the nucleotide sequences shown in SEQ ID NO: 112 and SEQ ID NO: 121 respectively encode the heavy and light chains of the 20C8 chimeric antibody, the nucleotide sequences shown in SEQ ID NO: 113 and SEQ ID NO: 122 respectively encode the heavy and light chains of the 23E12 chimeric antibody, the nucleotide sequences shown in SEQ ID NO: 114 and SEQ ID NO: 123 respectively encode the heavy and light chains of the 27H3 chimeric antibody, the nucleotide sequences shown in SEQ ID NO: 115 and SEQ ID NO: 124 respectively encode the heavy and light chains of the 21E5 chimeric antibody, and the nucleotide sequences shown in SEQ ID NO: 116 and SEQ ID NO: 125 respectively encode the heavy and light chains of the 21E5 chimeric antibody. The nucleotide sequences encode the heavy and light chains of the 2A5 chimeric antibody, the nucleotide sequences shown in SEQ ID NO:117 and SEQ ID NO:126 encode the heavy and light chains of the 4H4 chimeric antibody, the nucleotide sequences shown in SEQ ID NO:118 and SEQ ID NO:127 encode the heavy and light chains of the 22D12 chimeric antibody, the nucleotide sequences shown in SEQ ID NO:119 and SEQ ID NO:128 encode the heavy and light chains of the 1B9 chimeric antibody, and the nucleotide sequences shown in SEQ ID NO:120 and SEQ ID NO:129 encode the heavy and light chains of the 15D4 chimeric antibody, respectively. The nucleotide sequences shown in SEQ ID NO:130, SEQ ID NO:132, SEQ ID NO:134, SEQ ID NO:136, SEQ ID NO:138, SEQ ID NO:140, SEQ ID NO:142, SEQ ID NO:144, and SEQ ID NO:146 encode single chain antibody fragments of the nucleotide sequences shown in SEQ ID NO:94, SEQ ID NO:96, SEQ ID NO:98, SEQ ID NO:100, SEQ ID NO:102, SEQ ID NO:104, SEQ ID NO:106, SEQ ID NO:108, and SEQ ID NO:110, respectively, and the nucleotide sequences shown in SEQ ID NO:131, SEQ ID NO:133, SEQ ID NO:135, SEQ ID NO:137, SEQ ID NO:139, SEQ ID NO:141, SEQ ID NO:143, SEQ ID NO:145, and SEQ ID NO:147 encode single chain antibody fragments of the nucleotide sequences shown in SEQ ID NO:95, SEQ ID NO:97, SEQ ID NO:99, SEQ ID NO:101, SEQ ID NO:103, SEQ ID NO:105, SEQ ID NO:107, SEQ ID NO:109, and SEQ ID NO:111, respectively.

[0032] According to a third aspect of the present invention, the present invention provides an expression vector. According to an embodiment of the present invention, the expression vector carries the above-mentioned nucleic acid molecule. After the expression vector according to an embodiment of the present invention is introduced into a suitable recipient cell, the expression vector according to an embodiment of the present invention can effectively realize the expression of the above-mentioned antibody or antigen-binding fragment that specifically recognizes TrkA under the mediation of a regulatory system, thereby realizing the antibody or antigen-binding fragment to be obtained in large quantities in vitro.

[0033] According to an embodiment of the present invention, the expression vector may further comprise at least one of the following additional technical features: According to an embodiment of the present invention, the expression vector is a eukaryotic expression vector, thus achieving the expression of an antibody or an antigen-binding fragment thereof that specifically recognizes TrkA as described above in eukaryotic cells, such as CHO cells.

[0034] In a fourth aspect of the present invention, the present invention provides a recombinant cell. According to an embodiment of the present invention, the recombinant cell harbors the above-mentioned nucleic acid molecule or expresses the above-mentioned antibody or antigen-binding fragment thereof. The recombinant cell according to an embodiment of the present invention can be used for in vitro expression and large-scale production of the antibody or antigen-binding fragment thereof that specifically recognizes TrkA as described above.

[0035] According to an embodiment of the present invention, the above-mentioned recombinant cell may further include at least one of the following additional technical features: According to an embodiment of the present invention, the recombinant cell is obtained by introducing the above-mentioned expression vector into a host cell.

[0036] According to an embodiment of the present invention, the expression vector is introduced into the host cell by electrotransduction.

[0037] According to an embodiment of the present invention, the recombinant cell is a eukaryotic cell.

[0038] According to an embodiment of the present invention, the recombinant cell is a mammalian cell.

[0039] In the fifth aspect of the present invention, the present invention provides a pharmaceutical composition. According to an embodiment of the present invention, the pharmaceutical composition comprises the above-mentioned antibody, the above-mentioned nucleic acid molecule, the above-mentioned expression vector or the above-mentioned recombinant cell. The antibody or the expressed antibody contained in the pharmaceutical composition according to the embodiment of the present invention can specifically target and bind to TrkA receptor, block the binding of NGF and TrkA, and effectively inhibit pain, and also has the characteristics of low immunogenicity and almost no antibody-dependent cell-mediated cytotoxicity (ADCC).

[0040] In a sixth aspect of the invention, the invention provides the use of an antibody as defined above, a nucleic acid molecule as defined above, an expression vector as defined above or a recombinant cell as defined above, and a pharmaceutical composition as defined above in the preparation of a medicament for the treatment or prevention of pain, cancer, inflammation or inflammatory diseases, neurodegenerative diseases, Sjogren's syndrome, endometriosis, diabetic peripheral neuropathy, prostatitis, pelvic pain syndrome, diseases associated with imbalance in the regulation of bone remodelling and diseases caused by abnormal signalling of connective tissue growth factor.

[0041] According to an embodiment of the present invention, the above-mentioned use may further include at least one of the following additional technical features: According to an embodiment of the present invention, the medicament is used to treat or prevent neuropathic pain, inflammatory pain, cancer-related pain, fracture-related pain, surgery-related pain, inflammatory lung disease, interstitial cystitis, painful bladder syndrome, inflammatory bowel disease, inflammatory skin disease, Raynaud's syndrome, idiopathic pulmonary fibrosis, scarring (hypertrophic, keloid-type and other forms), sclerosis, endomyocardial fibrosis, atrial fibrosis, myelofibrosis, progressive massive fibrosis (lung), nephrogenic systemic fibrosis, scleroderma, systemic sclerosis, arthrofibrosis, ocular fibrosis, non-small cell lung cancer, papillary thyroid carcinoma, glioblastoma multiforme, colorectal cancer, melanoma, cholangiocarcinoma or sarcoma, acute myeloid leukemia, large cell neuroendocrine carcinoma, neuroblastoma, prostate cancer, pancreatic cancer, melanoma, squamous cell carcinoma of the head and neck or gastric cancer.

[0042] In a sixth aspect of the invention, the invention provides an antibody as defined above, a nucleic acid molecule as defined above, an expression vector as defined above or a recombinant cell as defined above, and a pharmaceutical composition as defined above for use in the preparation of a medicament for the treatment or prevention of pain, cancer, inflammation or an inflammatory disease, a neurodegenerative disease, Sjogren's syndrome, endometriosis, diabetic peripheral neuropathy, prostatitis, pelvic pain syndrome, diseases associated with imbalance in the regulation of bone remodelling and diseases caused by abnormal signalling of connective tissue growth factor.

[0043] According to an embodiment of the present invention, the above-mentioned use may further include at least one of the following additional technical features: According to an embodiment of the present invention, the medicament is used to treat or prevent neuropathic pain, inflammatory pain, cancer-related pain, fracture-related pain, surgery-related pain, inflammatory lung disease, interstitial cystitis, painful bladder syndrome, inflammatory bowel disease, inflammatory skin disease, Raynaud's syndrome, idiopathic pulmonary fibrosis, scarring (hypertrophic, keloid-type and other forms), sclerosis, endomyocardial fibrosis, atrial fibrosis, myelofibrosis, progressive massive fibrosis (lung), nephrogenic systemic fibrosis, scleroderma, systemic sclerosis, arthrofibrosis, ocular fibrosis, non-small cell lung cancer, papillary thyroid carcinoma, glioblastoma multiforme, colorectal cancer, melanoma, cholangiocarcinoma or sarcoma, acute myeloid leukemia, large cell neuroendocrine carcinoma, neuroblastoma, prostate cancer, pancreatic cancer, melanoma, squamous cell carcinoma of the head and neck or gastric cancer.

[0044] In a sixth aspect of the present invention, the present invention provides a method for treating or preventing pain, cancer, inflammation or inflammatory diseases, neurodegenerative diseases, Sjogren's syndrome, endometriosis, diabetic peripheral neuropathy, prostatitis, pelvic pain syndrome, diseases associated with imbalance in the regulation of bone remodeling and diseases caused by abnormal signaling of connective tissue growth factor in a subject, the method comprising the step of administering to a subject in need of such treatment or prevention a therapeutically effective amount of the above-mentioned antibody, the above-mentioned nucleic acid molecule, the above-mentioned expression vector or the above-mentioned recombinant cell, and the above-mentioned pharmaceutical composition.

[0045] According to an embodiment of the present invention, the above-mentioned method may further include at least one of the following additional technical features: According to an embodiment of the present invention, the medicament is used to treat or prevent neuropathic pain, inflammatory pain, cancer-related pain, fracture-related pain, surgery-related pain, inflammatory lung disease, interstitial cystitis, painful bladder syndrome, inflammatory bowel disease, inflammatory skin disease, Raynaud's syndrome, idiopathic pulmonary fibrosis, scarring (hypertrophic, keloid-type and other forms), sclerosis, endomyocardial fibrosis, atrial fibrosis, myelofibrosis, progressive massive fibrosis (lung), nephrogenic systemic fibrosis, scleroderma, systemic sclerosis, arthrofibrosis, ocular fibrosis, non-small cell lung cancer, papillary thyroid carcinoma, glioblastoma multiforme, colorectal cancer, melanoma, cholangiocarcinoma or sarcoma, acute myeloid leukemia, large cell neuroendocrine carcinoma, neuroblastoma, prostate cancer, pancreatic cancer, melanoma, head and neck squamous cell carcinoma or gastric cancer.

[0046] In a seventh aspect of the present invention, the present invention provides a kit for detecting TrkA. According to an embodiment of the present invention, the kit includes any one of the antibodies described above. The TrkA antibody described above can specifically target and bind to TrkA. The kit according to an embodiment of the present invention can realize specific detection of TrkA. For example, when the antibody is combined with a fluorescent group, a fluorescent detection device can be used to realize localization or real-time detection of TrkA.

[0047] In an eighth aspect of the invention, the present invention provides the use of an antibody as described above, a nucleic acid molecule as described above, an expression vector as described above, or a recombinant cell as described above in the preparation of a kit for detecting TrkA or diagnosing a TrkA-related disease.

[0048] In an eighth aspect of the present invention, the present invention provides an antibody as described above, a nucleic acid molecule as described above, an expression vector as described above, or a recombinant cell as described above for use in the preparation of a kit for detecting TrkA or diagnosing a TrkA-related disease.

[0049] In an eighth aspect of the present invention, the present invention provides a method for detecting TrkA or diagnosing a TrkA-related disease in a subject, the method comprising the step of administering to a subject in need of detection or diagnosis a therapeutically effective amount of the above-mentioned antibody, the above-mentioned nucleic acid molecule, the above-mentioned expression vector, or the above-mentioned recombinant cell.

[0050] In a ninth aspect of the invention, the invention provides a murine B cell. According to an embodiment of the invention, the genome of the B cell carries a sequence encoding a constant region, the constant region sequence having S108P, F114A, L115A, R289K mutations and a 327K deletion mutation compared to the constant region of human IgG4. The constant region of human IgG4 has the amino acid sequence shown in SEQ ID NO: 148. ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 148)

[0051] Mouse B cells according to embodiments of the invention can be used to secrete mouse antibodies whose immunogenicity in humans is further reduced, whose stability is significantly improved, and whose half-life in vivo is significantly increased.

[0052] In a tenth aspect of the invention, the invention provides the use of a murine B cell as described above in the preparation of a monoclonal antibody.

[0053] In a tenth aspect of the invention, the invention provides a murine B cell as described above for use in the preparation of a monoclonal antibody.

[0054] In a tenth aspect of the invention, the invention provides a method for detecting TrkA or diagnosing a TrkA-related disease, comprising culturing mouse B cells as described above to produce monoclonal antibodies. [Brief description of the drawings]

[0055] [Figure 1] FIG. 1 shows experimental results of nine monoclonal hybridoma cell lines capable of producing anti-TrkA monoclonal antibodies that block the binding of NGF and TrkA at the molecular level, obtained by screening with an ELISA method according to an embodiment of the present invention. [Diagram 2] FIG. 1 is a diagram of experimental results (% parent) of nine monoclonal hybridoma cell lines obtained by flow screening according to an embodiment of the present invention, capable of producing anti-TrkA monoclonal antibodies that block the binding of NGF and TrkA at the cellular level. [Diagram 3] FIG. 1 shows the experimental results of detecting the binding of monoclonal antibodies produced by each positive clone supernatant to mouse TrKA receptor by ELISA method according to an embodiment of the present invention. [Figure 4] FIG. 1 shows experimental results of evaluation of blocking activity of test antibodies using a HEK-293T-TrkA cell model (% parent), according to an embodiment of the present invention. [Diagram 5] FIG. 1 shows experimental results of tanezumab used to verify whether the NIH-3T3-TrkA cell model can be used to evaluate the activity of drugs (test drugs) in vitro that inhibit the NGF-TrkA pathway, according to an embodiment of the present invention. [Figure 6]FIG. 1 shows experimental results of tyrosine phosphorylation levels of TrkA protein under the action of test antibodies and positive antibody MNAC13 by AlphaLISA method according to an embodiment of the present invention. [Figure 7] FIG. 1 shows the results of detecting the affinity EC50 of test antibodies by flow cytometry according to an embodiment of the present invention. [Figure 8AB] FIG. 1 shows the results of evaluating the specificity of binding of test antibodies to the target TrKA by ELISA according to an embodiment of the present invention. [Figure 8C-D] FIG. 1 shows the results of evaluating the specificity of binding of test antibodies to the target TrKA by ELISA according to an embodiment of the present invention. [Fig. 8EF] FIG. 1 shows the results of evaluating the specificity of binding of test antibodies to the target TrKA by ELISA according to an embodiment of the present invention. [Figure 8G] FIG. 1 shows the results of evaluating the specificity of binding of test antibodies to the target TrKA by ELISA according to an embodiment of the present invention. [Figure 9] FIG. 1 shows the results of detecting the binding ability of a test antibody to mouse-TrKA protein by providing an ELISA method according to an embodiment of the present invention. [Figure 10] FIG. 1 shows the results of detecting the binding ability of a test antibody to mouse-TrKA protein by flow cytometry according to an embodiment of the present invention. [Figure 11] FIG. 1 shows the results of detecting the inhibitory effect of test antibodies on the binding of mouse-NGF and mouse-TrKA by applying an ELISA method according to an embodiment of the present invention. [Figure 12] FIG. 1 shows the results of detecting the inhibitory effect of test antibodies on the binding of mouse-NGF and mouse-TrKA by flow cytometry according to an embodiment of the present invention. [Figure 13] FIG. 1 shows the results of detecting the inhibitory effect of test antibodies on the binding of mouse-NGF and mouse-TrKA by applying an ELISA method according to an embodiment of the present invention. [Figure 14]FIG. 1 shows results of evaluating the analgesic activity of test antibodies in vivo using a formalin pain model, according to an embodiment of the present invention. [Figure 15] FIG. 1 shows results of evaluating the analgesic activity of test antibodies in vivo using a Freund's adjuvant-induced inflammation and pain model, according to an embodiment of the invention. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0056] DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS The embodiments of the present invention are described in detail below. Examples of the embodiments are illustrated in the accompanying drawings, in which the same or similar reference numerals represent the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the drawings are exemplary and are intended to explain the present invention, but should not be construed as limiting the present invention.

[0057] In the course of describing the present invention, the terms used herein are explained. These explanations are merely for the convenience of understanding the concept, and should not be considered as limiting the protection concept of the present invention.

[0058] antibody As used herein, the term "antibody" refers to an immunoglobulin molecule capable of binding to a specific antigen. An antibody is composed of two lighter molecular weight light chains and two heavier molecular weight heavy chains. The heavy (H) chain and the light (L) chain are linked by disulfide bonds to form a tetrapeptide chain molecule. Among them, the amino acid sequence at the amino terminal (N-terminal) of the peptide chain is highly variable, which is called the variable region (V region). The carboxyl terminal (C-terminal) is relatively stable with little change, which is called the constant region (C region). The V regions of the L chain and the H chain are called VL and VH, respectively.

[0059] Some regions in the variable region show a higher degree of variation in amino acid composition and sequence. They are called hypervariable regions (HVRs). Hypervariable regions are where antigens and antibodies bind, and therefore they are also called complementarity determining regions (CDRs). There are three CDRs on both the heavy and light chain variable regions.

[0060] The present invention utilizes the extracellular segment of TrkA to obtain highly specific, high affinity anti-TrkA Fab (fragment antigen binding) antibody fragments by immunization, which can specifically bind to the TrkA antigen and thus can be used for targeted treatment of diseases such as pain or tumors.

[0061] In some embodiments, the present invention provides an antibody or antigen-binding fragment capable of specifically recognizing TrkA, the antibody comprising a CDR sequence selected from at least one of the following: heavy chain variable region CDR sequences: SEQ ID NO: 1 to SEQ ID NO: 27, light chain variable region CDR sequences: SEQ ID NO: 28 to SEQ ID NO: 54, or an amino acid sequence having at least 95% identity thereto. In other embodiments, the antibody or antigen-binding fragment provided by the present invention has conservative amino acid substitutions compared to the heavy and light chains. An "antigen-binding fragment" refers to an antibody fragment that retains the ability to specifically bind to an antigen (ROR2). A "conservative amino acid substitution" refers to the replacement of an amino acid with a residue that is biologically, chemically, or structurally similar to another amino acid. Biological similarity means that the substitution does not destroy the biological activity of the TrkA antibody or TrkA antigen. Structural similarity refers to side chains with amino acids of similar length, such as alanine, glycine, or serine, or side chains of similar size. Chemical similarity means that the amino acids have the same charge or are both hydrophilic or hydrophobic. For example, the hydrophobic residues isoleucine, valine, leucine, or methionine are substituted for each other. Alternatively, a polar amino acid is substituted for another polar amino acid, e.g., lysine is substituted for arginine, aspartic acid is substituted for glutamic acid, asparagine is substituted for glutamine, threonine is substituted for serine, etc.

[0062] In some embodiments, the present invention provides an antibody or antigen-binding fragment having a heavy chain variable region of an amino acid sequence shown in any one of SEQ ID NOs: 55 to 63 and a light chain variable region of an amino acid sequence shown in any one of SEQ ID NOs: 64 to 72. The inventors can obtain the CDR regions of the above-mentioned anti-heavy chain variable region sequences (as shown in SEQ ID NOs: 1 to 27) and the CDR regions of the light chain variable region sequences (such as SEQ ID NOs: 28 to 54) from antibody sequence alignment databases (NCBI, IMGT). In other embodiments, the heavy chain variable region sequence of the antibody or antigen-binding fragment has conservative amino acid substitutions compared to the amino acid sequence shown in SEQ ID NOs: 55 to 63. In some embodiments, the light chain variable region sequence of the antibody or antigen-binding fragment has conservative amino acid substitutions compared to the amino acid sequence shown in any one of SEQ ID NOs: 64 to 72. Naturally, these conservative amino acid substitutions do not change the biological function of the antibody or antigen-binding fragment. In some particular modes, these conservative amino acid substitutions may occur at amino acids outside the CDR regions in the heavy and light chain variable regions.

[0063] In some preferred embodiments, the present invention provides anti-TrkA antibodies having a heavy chain with an amino acid sequence shown in any one of SEQ ID NOs: 76 to 84, and a light chain with an amino acid sequence shown in any one of SEQ ID NOs: 85 to 93.

[0064] In some preferred embodiments, the invention provides anti-TrkA single chain fragment antibodies, wherein the antibodies have the amino acid sequences set forth in SEQ ID NO:94 to SEQ ID NO:111.

[0065] Nucleic acid molecules, expression vectors, and recombinant cells In the process of preparing or obtaining these antibodies, the nucleic acid molecules expressing these antibodies can be ligated into various vectors and then expressed in various cells to obtain the corresponding antibodies.

[0066] To this end, the present invention also provides isolated nucleic acid molecules encoding the above-described antibodies or antigen-binding fragments.

[0067] In some embodiments, the isolated nucleic acid molecule has a nucleotide sequence set forth in any one of SEQ ID NOs: 112 to 120, or has a nucleotide sequence set forth in any one of SEQ ID NOs: 121 to 129, or has a nucleotide sequence set forth in any one of SEQ ID NOs: 130 to 147.

[0068] In some embodiments, the isolated nucleic acid molecule has at least 90% homology, preferably at least 95% homology, more preferably at least 98%, 99% homology, with the nucleotide sequence shown in SEQ ID NO: 112 to SEQ ID NO: 120. In at least some embodiments, the isolated polynucleotide has at least 90% homology, preferably at least 95% homology, more preferably at least 98%, 99% homology, with the nucleotide sequence shown in SEQ ID NO: 121 to SEQ ID NO: 129. In at least some embodiments, the isolated polynucleotide has at least 90% homology, preferably at least 95% homology, more preferably at least 98%, 99% homology, with the nucleotide sequence shown in SEQ ID NO: 130 to SEQ ID NO: 147. These sequences homologous to the nucleotide sequences shown in SEQ ID NO:112 to SEQ ID NO:120 or SEQ ID NO:121 to SEQ ID NO:129 or SEQ ID NO:130 to SEQ ID NO:147 can express amino acids similar to SEQ ID NO:76 to SEQ ID NO:84 or SEQ ID NO:85 to SEQ ID NO:93 or SEQ ID NO:94 to SEQ ID NO:111, so that they can specifically bind to the TrkA antigen and achieve the targeting function of the antibody.

[0069] In some preferred embodiments, the isolated nucleic acid molecule has a heavy chain nucleotide sequence set forth in SEQ ID NO: 112 to SEQ ID NO: 120 and a light chain nucleotide sequence set forth in SEQ ID NO: 121 to SEQ ID NO: 129. These nucleotide sequences are optimized for the species and are more readily expressed in mammalian cells.

[0070] The present invention also provides an expression vector comprising the isolated nucleic acid molecule described above. When the isolated polypeptide described above is ligated to a vector, the polynucleotide can be directly or indirectly linked to control elements on the vector, so long as these control elements can control the translation and expression of the polynucleotide. Of course, these control elements can be directly derived from the vector itself, or these control elements can be exogenous, i.e., not derived from the vector itself. Of course, the polynucleotide can be operably linked to the control elements. "Operably linked" in this specification refers to the binding of an exogenous gene to a vector, so that the control elements in the vector, such as transcriptional and translational control sequences, can perform their expected function of regulating the transcription and translation of the exogenous gene. Of course, the polynucleotides used to code for the heavy and light chains of the antibody can be inserted independently into different vectors, and they are usually inserted into the same vector. Commonly used vectors can be, for example, plasmids, phages, etc. For example, Plasmid-X plasmid.

[0071] The present invention also provides recombinant cells containing the expression vector. The expression vector can be introduced into mammalian cells, recombinant cells can be constructed and obtained, and these recombinant cells can then be used to express the antibodies or antigen-binding fragments provided by the present invention. By culturing the recombinant cells, the corresponding antibodies can be obtained. These usable mammalian cells can be, for example, CHO cells, etc.

[0072] Pharmaceutical compositions, kits and pharmaceutical uses and uses in the preparation of kits The present invention also provides a pharmaceutical composition comprising the antibody or antigen-binding fragment described above and a pharma- ceutically acceptable carrier.

[0073] The anti-TrkA antibodies provided herein may be incorporated into pharmaceutical compositions suitable for administration to a subject. In general, these pharmaceutical compositions include the anti-TrkA antibodies provided herein and a pharma- ceutically acceptable carrier. A "pharmaceutically acceptable carrier" may include any and all physiologically compatible solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption-retarding agents, and the like. Specific examples may be one or more of water, saline, phosphate buffered saline, glucose, glycerol, ethanol, and the like, and combinations thereof. In many cases, pharmaceutical compositions include isotonic agents, such as sugars, polyalcohols (such as mannitol, sorbitol, or sodium chloride. Of course, a pharma- ceutically acceptable carrier may also include minor amounts of auxiliary substances, such as wetting or emulsifying agents, preservatives, or buffers, to extend the shelf life or effectiveness of the antibody.

[0074] For example, the antibodies of the present invention can be incorporated into pharmaceutical compositions suitable for parenteral administration (e.g., intravenous, subcutaneous, intraperitoneal, intramuscular). These pharmaceutical compositions can be prepared in various forms. Examples are liquid, semi-solid and solid dosage forms, including, but not limited to, liquid solutions (e.g., injection and infusion solutions), dispersions or suspensions, tablets, pills, powders, liposomes, and suppositories. Typical pharmaceutical compositions are in the form of injection or infusion solutions. The antibodies can be administered by intravenous infusion or injection or intramuscular or subcutaneous injection.

[0075] Of course, the anti-TrkA antibodies herein can also be part of a kit or other diagnostic reagent, if desired. According to an embodiment of the present invention, the present invention also provides a kit comprising the above-mentioned TrkA antibodies. The kits provided by the present invention can be used for detection, for example, by immunoblotting, immunoprecipitation, etc., including using the specific binding properties of the TrkA antigen and the antibody. These kits can include any one or more of the following: antagonist, anti-TrkA antibody or drug reference material, protein purification columns, immunoglobulin affinity purification buffer, cell assay diluent, instructions or literature, etc. The anti-TrkA antibodies can be used in various types of diagnostic tests, such as detecting various diseases or the presence of drugs, toxins or other proteins, in vitro or in vivo. For example, the anti-TrkA antibodies can be used to test for related diseases by detecting the serum or blood of a subject. Such associated diseases may include pain, cancer, inflammation or inflammatory diseases, neurodegenerative diseases, Sjogren's syndrome, endometriosis, diabetic peripheral neuropathy, prostatitis, pelvic pain syndrome, diseases associated with imbalance in the regulation of bone remodeling, and TrkA-associated diseases such as diseases caused by abnormal signaling of connective tissue growth factor. Of course, the antibodies provided herein may also be used for radioimmunodetection and radioimmunotherapy of the above diseases.

[0076] Specifically, the above-mentioned pain, inflammation or inflammatory disease, neurodegenerative disease, Sjogren's syndrome, endometriosis, diabetic peripheral neuropathy, prostatitis, pelvic pain syndrome, diseases associated with imbalance in the regulation of bone remodeling, and diseases caused by abnormal signaling of connective tissue growth factor include neuropathic pain, inflammatory pain, cancer-related pain, fracture-related pain, surgery-related pain, inflammatory lung disease, interstitial cystitis, painful bladder syndrome, inflammatory bowel disease, inflammatory skin disease, Raynaud's syndrome, idiopathic pulmonary fibrosis, scarring (hypertrophy, keloid type and other forms), sclerosis, endomyocardial fibrosis, atrial fibrosis, myelofibrosis, progressive massive fibrosis (lung), nephrogenic systemic fibrosis, scleroderma, systemic sclerosis, arthrofibrosis, and ocular fibrosis.

[0077] These cancers or tumors can be any uncontrolled cell growth, such as non-small cell lung cancer, papillary thyroid carcinoma, glioblastoma multiforme, colorectal cancer, melanoma, cholangiocarcinoma or sarcoma, acute myeloid leukemia, large cell neuroendocrine carcinoma, neuroblastoma, prostate cancer, pancreatic cancer, melanoma, squamous cell carcinoma of the head and neck, or gastric cancer.

[0078] When the anti-TrkA antibody provided by the present invention is used to treat the above-mentioned diseases, the anti-TrkA antibody provided by the present invention can be provided to a subject. To this end, the present invention provides a method for treating the above-mentioned diseases, comprising administering the antibody or antigen-binding fragment thereof provided by the present invention to a subject in need thereof.

[0079] The solution of the present invention is described below with reference to examples. Those skilled in the art will understand that the following examples are merely used to illustrate the present invention and should not be considered as limiting the scope of the present invention. If a specific technique or condition is not shown in the examples, the technique or condition described in the literature or product description in the art will be carried out. If the reagents or equipment used are not specified by the manufacturer, they are all conventional products available on the market. EXAMPLES

[0080] Screening of anti-TrkA monoclonal antibodies by hybridoma technology. Hybridoma technology and ELISA method were used to screen hybridoma cell lines that could produce anti-TrkA monoclonal antibodies that block the binding of NGF and TrkA at the molecular level. Using the NCBI database, a gene encoding the extracellular domain of TrkA protein was designed, which was constructed in a mammalian eukaryotic expression system. The extracellular domain of TrkA protein was expressed and used for screening anti-TrkA monoclonal antibodies. The extracellular domain of immunogen TrkA protein was obtained from Beijing Yiqiao Shenzhou Biotechnology Co., Ltd. Balb / C mice were immunized three times by intraperitoneal injection, and the serum antibody titers of the immunized mice were determined by ELISA method and were 10 5After boosting the immunity by splenic injection, it was fused with myeloma cells. HAT and HT selection media were used as selection media and to screen the fused hybridoma cell lines, ELISA method was used to screen the positive hybridoma cell lines capable of producing anti-TrkA antibody, and after performing clonal culture of the positive hybridoma cells by limiting dilution method, stable hybridoma cell lines capable of producing anti-TrkA monoclonal antibody were selected. After three cloning cultures and four ELISA tests, a total of 147 positive monoclonal hybridoma cell lines were screened. NGF was biotinylated. NGF could bind to the extracellular domain of TrkA protein, and anti-TrkA monoclonal antibody could also bind to the extracellular domain of TrkA protein. A competitive experiment was designed to detect the binding of NGF to the extracellular domain of TrkA protein under the action of anti-TrkA monoclonal antibody by ELISA, and to screen for anti-TrkA monoclonal antibody that blocks the binding of NGF and TrkA at the molecular level. Figure 1 shows the results of the blocking experiment of 9 hybridoma cell lines that can produce anti-TrkA monoclonal antibody that blocks the binding of NGF and TrkA at the molecular level among 147 positive monoclonal hybridoma cell lines, that is, a total of 9 monoclonal hybridoma cell lines that can produce anti-TrkA monoclonal antibody that blocks the binding of NGF and TrkA at the molecular level are obtained. The experimental results are shown in Figure 1. In Figure 1, OD 450reflects the NGF signal binding to the extracellular domain of TrkA protein. The higher the reading, the stronger the NGF signal binding to the extracellular domain of TrkA protein, and the antibody is less effective in blocking NGF binding to TrkA. As shown in FIG. 1, compared with the negative clone group, the ELISA reading of the 9 positive clone group is significantly reduced, and the NGF signal binding to the extracellular domain of TrkA protein is significantly reduced, indicating that these 9 positive clones can reduce the NGF signal binding to the extracellular domain of TrkA protein by generating anti-TrkA monoclonal antibody that blocks NGF binding to the extracellular domain of TrkA protein. EXAMPLES

[0081] Screening of anti-TrkA monoclonal antibodies using HEK-293T-TrkA cell model Using lentivirus technology, we constructed a HEK-293T-TrkA cell model and screened the hybridoma cell line capable of producing anti-TrkA monoclonal antibody that blocks the binding of NGF and TrkA at the cellular level by flow cytometry. NGF was biotinylated, so that NGF could bind to the extracellular domain of TrkA protein on HEK-293T-TrkA cells, and anti-TrkA monoclonal antibody could also bind to the extracellular domain of TrkA protein on HEK-293T-TrkA cells. A competition experiment was designed to detect the binding of NGF to the extracellular domain of TrkA protein on HEK-293T-TrkA cells under the action of anti-TrkA monoclonal antibody by flow cytometry, and to screen the anti-TrkA monoclonal antibody that blocks the binding of NGF and TrkA at the cellular level. A total of nine monoclonal hybridoma cell lines capable of producing anti-TrkA monoclonal antibodies that block the binding of NGF and TrkA at the cellular level were obtained by screening, which are consistent with the results of the molecular level blocking experiment. The experimental results are shown in Figure 2. In Figure 2, the parental % value reflects the NGF signal binding to the extracellular domain of TrkA protein on HEK-293T-TrKA cells. The higher the reading, the stronger the NGF signal binding to the extracellular domain of TrkA protein on HEK-293T-TrKA cells, and the worse the effect of the antibody in blocking NGF binding to TrkA. As shown in FIG. 2, compared with the negative clone group, the parent % value of the 9 positive clone group was significantly reduced, and the NGF signal binding to the extracellular domain of TrkA protein was significantly reduced. It can be seen that by generating anti-TrkA monoclonal antibody that blocks NGF binding to the extracellular domain of TrkA protein on HEK-293T-TrkA cells, these 9 positive clones can reduce the NGF signal binding to the extracellular domain of TrkA protein on HEK-293T-TrkA cells. EXAMPLES

[0082] Detection of human-mouse cross-reactivity of positive clones producing monoclonal antibodies by ELISA method The binding of the monoclonal antibodies produced by the supernatant of each positive clone to the mouse TrKA receptor was detected by ELISA, and the human-mouse cross-reactivity of the positive clones producing the monoclonal antibodies was detected. The results are shown in Figure 3. In Figure 3, the OD 450 The value reflects the strength of the antibody's binding to the protein. The higher the reading, the stronger the antibody's binding to the protein. As shown in FIG. 3, the OD of the 23E12, 21E5, 15D4, and 27H3 groups 450 The OD values ​​were approximately 3.5, which was significantly different from the negative control group and the OD values ​​of the 20C8, 1B9, 4H4, 3A5, and 22D12 groups. 450 The values ​​were almost 0, and no significant difference was observed compared to the negative clone group. Clones 23E12, 21E5, 15D4, and 27H3 were able to produce monoclonal antibodies that bound to both human-TrKA and mouse-TrKA, indicating the presence of human-mouse cross-reactivity. EXAMPLES

[0083] Vector construction Using molecular cloning techniques, a series of (20C8, 23E12, 27H3, 21E5, 2A5, 4H4, 1B9, 22D12, 15D4) chimeric antibody expression vectors were constructed. The chimeric antibodies were recombinantly expressed in a CHO expression system. The nucleotide sequences encoding the light and heavy chains of a series of (20C8, 23E12, 27H3, 21E5, 2A5, 4H4, 1B9, 22D12, 15D4) chimeric monoclonal antibodies were obtained by chemical synthesis by Jinweizhi Biotechnology Co., Ltd. The obtained sequences were double digested and inserted into the same digestion site of a eukaryotic expression vector to construct a series of (20C8, 23E12, 27H3, 21E5, 2A5, 4H4, 1B9, 22D12, 15D4) chimeric antibody expression vectors. A series of verified correct expression vectors were then extracted with an Invitrogen plasmid extraction kit, linearized with restriction enzymes, purified, harvested, and stored at -20°C. EXAMPLES

[0084] Transfection of vectors encoding a series of chimeric antibodies and expression in cells After the CHO host cells were resuscitated in CD CHO medium, the cell density reached approximately 8 * 10 5 Once cells / mL were obtained, the cells were harvested for transfection. Transfected cells were approximately 1 * 10 7 The total number of cells was approximately 100, and the vector was approximately 40 μg. The cells were transfected by electric shock (Bio-Rad, Gene pulser Xcell). After electric shock, the cells were cultured in 20 mL of CD CHO medium. The day after culture, the cells were harvested by centrifugation and resuspended in 20 mL of CD CHO medium containing MSX to a final concentration of 50 μM. The cell density was approximately 0.6 * 10 6 When the cell density reached 100 cells / mL, the obtained mixed clones were subcultured using CD CHO medium to a subculture cell density of approximately 0.2 * 10 6 The cell culture solution was harvested when the cell viability was approximately 90%. EXAMPLES

[0085] Purification of chimeric antibodies by harvesting cell fermentation medium A series of chimeric antibodies were examined at the translation level. A Protein A chromatography column was used to purify the collected cell culture solution, and the absorption peak was collected for mass spectrometry detection. Mass spectrometry detected a series of chimeric antibodies with molecular weights of about 150 KD, which was consistent with the theoretical molecular weight. At the same time, the collected samples were detected by 10% SDS-PAGE electrophoresis after reduction and non-reduction. The reduced SDS-PAGE electrophoresis spectrum showed two bands at about 25 KD and 50 KD, respectively. The non-reduced SDS-PAGE electrophoresis spectrum showed a single band at about 150 KD. The band sizes of the electrophoresis spectrum were consistent with the theory. The purified samples were dialyzed overnight at 4°C against 0.02 M PBS buffer at pH 7.4.

[0086] In Examples 7 to 16 below, the inventors evaluated the constructed and purified chimeric antibodies 20C8, 23E12, 27H3, 21E5, 2A5, 4H4, 1B9, 22D12, and 15D4 for their affinity to TrkA and their activity in binding to TrkA and blocking of NGF to the extracellular domain of the TrkA protein. EXAMPLES

[0087] Evaluation of the blocking activity of test antibodies using the HEK-293T-TrkA cell model NGF was biotinylated, and NGF could bind to the extracellular domain of TrkA protein on HEK-293T-TrkA cells, and anti-TrkA monoclonal antibody could also bind to the extracellular domain of TrkA protein on HEK-293T-TrkA cells. A competition experiment was designed to detect the binding of NGF to the extracellular domain of TrkA protein on HEK-293T-TrkA cells under various concentrations (20 μg / mL, 10 μg / mL, 5 μg / mL, 2.5 μg / mL, 1.25 μg / mL, 0.625 μg / mL, 0.313 μg / mL, 0.156 μg / mL, 0.078 μg / mL, 0.039 μg / mL, 0.019 μg / mL) of the test antibody by flow cytometry, and to study the inhibitory effect of the test antibody on the binding of NGF and TrKA. The experimental results are shown in Figure 4. In FIG. 4, the parental % value reflects the NGF signal binding to the extracellular domain of TrKA protein on HEK-293T-TrkA cells. The lower the reading, the weaker the NGF signal binding to the extracellular domain of TrkA protein on HEK-293T-TrkA cells, and the greater the effect of the antibody on inhibiting the binding of NGF to TrKA; as shown in FIG. 4, with increasing test antibody concentration, the parental % value gradually decreases until it approaches zero, i.e., the NGF signal binding to the extracellular domain of TrkA protein gradually decreases until there is no NGF binding to the extracellular domain of TrkA protein and all of the binding of NGF to TrkA is inhibited. It can be seen that within a certain concentration range, each of the test antibodies can dose-dependently inhibit the binding of NGF to TrkA at the cellular level. EXAMPLES

[0088] Evaluating the activity of test antibodies in vitro using the NIH-3T3-TrkA cell model Under NGF stimulation, the tyrosine phosphorylation level of TrkA protein on the membrane of NIH-3T3-TrkA cells was up-regulated, and the TrkA downstream signaling pathway was activated. The test antibody could bind to the TrkA protein on the surface of the NIH-3T3-TrkA cell membrane, inhibit NGF stimulation, and down-regulate the tyrosine phosphorylation level of TrkA protein. In the test, IgG4 was used as a negative control (the antibody does not bind to NGF and TrkA), and tanezumab (anti-NGF monoclonal antibody, the amino acid sequence of the light chain is shown in SEQ ID NO: 149, the amino acid sequence of the heavy chain is shown in SEQ ID NO: 150, a drug that inhibits the NGF-TrkA pathway) and MNAC13 (anti-TrKA monoclonal antibody, the amino acid sequence of the light chain is shown in SEQ ID NO: 151, the amino acid sequence of the heavy chain is shown in SEQ ID NO: 152, a drug that inhibits the NGF-TrkA pathway) were used as positive controls. Among them, Tanezumab was used to verify whether the NIH-3T3-TrkA cell model can be used to evaluate the in vitro activity of drugs (test drugs) that inhibit the NGF-TrkA pathway. The experimental data are shown in Figure 5. The AlphaLISA method was used to detect the downregulation of tyrosine phosphorylation of TrkA protein under the action of various concentrations of the test antibody and the positive antibody MNAC13 to measure the activity of the test antibody in vitro. The test results of p-TrkA are shown in Figure 6. The experimental results show that both the test antibody and the positive antibody MNAC13 can inhibit the NGF-TrKA pathway and downregulate the tyrosine phosphorylation level of TrkA protein in a dose-dependent manner. Among them, the half inhibitory concentrations IC of the test antibodies 23E12, 20C8, 27H3, 21E5, 1B9, and 4H4 were 50 is the half inhibitory concentration IC of the positive antibody MNAC13 50 It can be seen that the tested antibodies 23E12, 20C8, 27H3, 21E5, 1B9, and 4H4 have better activity in vitro than the positive antibody MNAC13. DIQMTQSPSSLSASVGDRVTITCRASQSISNNLNWYQQKPGKAPKLLIYYTSRFHSGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQEHTLPYTFGQGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 149) QVQLQESGPGLVKPSETLSLTCTVSGFSLIGYDLNWIRQPPGKGLEWIGIIWGDGTTDYNSAVKSRVTISKDTSKNQFSLKLSSVTAADTAVYYCARGGYWYATSYYFDYWGQG TLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECP PCPAPPVAGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNGKEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO: 150) DIVLTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKPGQAPKLLIYTTSNLASGVPSRFSGSGSGTDYTLTISSLQPEDVATYYCHQWSSYPWTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 151) EVQLLESGGGLVQPGGSLRLSCAASGFTFSTYTMSWARQAPGKGLEWVAYISKGGGSTYYPDTVKGRFTISRDNSKNTLYLQMNSLRAEDSAVYYCARGAMFGNDFFFPMDRWG QGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPC PPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG (SEQ ID NO: 152) EXAMPLES

[0089] Evaluation of the affinity of test antibodies to TrkA using the Fortebio method Test antibody (23E12, 20C8, 21E5, 27H3, 1B9, 4H4, 2A5) samples were diluted to 4 concentration gradients (80nM, 26.67nM, 8.89nM, 2.93nM) in buffer solution (100ml PBS buffer, add 0.1g BSA, stir until completely dissolved, then add 20μL Tween 20, mix thoroughly). They were added to each well of the sample plate. The affinity detection system (OCTET RED 96 SYSTEM) automatically detected the binding of TRKA protein with each test antibody after TRKA (50μg / ml, 75kDa, provided by HEC LabS) protein with histidine tag bound to Ni-NTA sensor (manufacturer: PALL, product code: 18-5101). The results are shown in Table 1. D The value is the equilibrium dissociation constant between an antibody and its antigen, K D The value is inversely proportional to the affinity.D The K value is related to the antibody concentration (the amount of antibody needed for a particular experiment). D The lower the value (lower concentration), the higher the affinity of the antibody. High affinity antibodies are generally in the low nanomolar range (10 -9 ) are thought to be present. The results in Table 1 show that the K D Values ​​are in the low nanomolar range (10 -9 ), indicating that the test antibody has high affinity.

[0090] [Table 1] EXAMPLES

[0091] Assessment of test antibody affinity by flow cytometry Test antibody (23E12, 20C8, 21E5, 27H3, 1B9, 4H4, 2A5) samples were diluted in PBS buffer solution to 11 concentration gradients (20 μg / mL, 10 μg / mL, 5 μg / mL, 2.5 μg / mL, 1.25 μg / mL, 0.625 μg / mL, 0.313 μg / mL, 0.156 μg / mL, 0.078 μg / mL, 0.039 μg / mL, 0.019 μg / mL). Flow cytometry was used to detect the binding of various concentration gradients of the test antibodies to the TrKA receptor on the surface of HEK-293-TrKA cells to evaluate the affinity of the test antibodies at the cellular level. The results are shown in Figure 7. In Figure 7, the EC 50 The EC (half binding concentration) value reflects the affinity of the antibody. 50 The smaller the value, the higher the affinity of the antibody. In general, the EC of a high affinity antibody 50 The EC values ​​are considered to be less than 1.5 μg / mL. The results in FIG. 7 show that the EC values ​​for the tested antibodies 23E12, 20C8, 21E5, 27H3, 1B9, 4H4, and 2A5 are 50 All values ​​were shown to be below 1.5 μg / mL, indicating that the tested antibodies have high affinity. EXAMPLES

[0092] Assessment of the specificity of the binding of the test antibody to the target TrKA by ELISA The TrkA receptor family belongs to the receptor tyrosine kinase (RTK) including TrkA, TrkB, and TrkC, which have high homology. TrkA is a receptor tyrosine kinase of nerve growth factor (NGF) that selectively binds to NGF and is the functional receptor for NGF. Besides the high affinity receptor TrkA, NGF can also bind to its low affinity receptor p75. In the study, the binding of various concentrations (20μg / mL, 10μg / mL, 5μg / mL, 2.5μg / mL, 1.25μg / mL, 0.625μg / mL, 0.313μg / mL, 0.156μg / mL, 0.078μg / mL, 0.039μg / mL, 0.019μg / mL) of the test antibodies to TrKA, TrKB, TrKC, and p75, respectively, was detected by ELISA to evaluate the specificity of the test antibodies binding to the target TrKA. The results are shown in Figure 8. In Figure 8, at a particular antibody concentration, the OD 450 The value reflects the binding strength of the antibody to the protein. The larger the reading, the stronger the binding of the antibody to the protein. Experimental results show that the tested antibodies 23E12, 20C8, 27H3, 21E5, 1B9, 4H4, and 2A5 all have good binding to the TrKA receptor (the concentration of the tested antibodies was increased from 0 μg / mL to 20 μg / mL, and the OD 450 The OD value gradually increased until it stabilized, which was close to 3), but did not bind to TrKB, KrKC, or P75 (the concentration of each test antibody was increased from 0 μg / mL to 20 μg / mL, and the OD 450 It is shown that the value remains almost unchanged, which is close to 0. It can be seen that the specificity of the binding of the test antibody to the target TrKA is very good. EXAMPLES

[0093] Detection of the binding ability of the test antibody to mouse TrKA protein by ELISA The test antibody (23E12, 21E5) samples were diluted with PBS buffer solution to 11 concentration gradients (20 μg / mL, 10 μg / mL, 5 μg / mL, 2.5 μg / mL, 1.25 μg / mL, 0.625 μg / mL, 0.313 μg / mL, 0.156 μg / mL, 0.078 μg / mL, 0.039 μg / mL, 0.019 μg / mL). The binding of the test antibody to the mouse-TrKA receptor of each concentration gradient was detected by ELISA, thus detecting the binding ability of the test antibody to the mouse-TrKA protein. The results are shown in Figure 9. In Figure 9, the OD 450 The value reflects the binding strength of the antibody to the protein. The larger the reading, the stronger the binding of the antibody to the protein. According to the experimental results, the concentration of the test antibody was increased from 0 μg / mL to 20 μg / mL, and the OD 450 The value is shown to increase gradually until it approaches a plateau, which is close to about 3.5. It can be seen that the tested antibodies 23E12, 21E5, and the mouse-TrKA protein all have good binding capacity. EXAMPLES

[0094] Detection of the binding ability of the test antibody to mouse-TrKA protein by flow cytometry The test antibody 23E12 sample was diluted with PBS buffer solution to 11 concentration gradients (20 μg / mL, 10 μg / mL, 5 μg / mL, 2.5 μg / mL, 1.25 μg / mL, 0.625 μg / mL, 0.313 μg / mL, 0.156 μg / mL, 0.078 μg / mL, 0.039 μg / mL, 0.019 μg / mL), and the binding of the test antibody to the mouse-TrKA receptor on the surface of HEK293T-mouse-TrKA cells at each concentration gradient was detected by flow cytometry. The antibody was tested for its ability to bind to the mouse-TrKA protein. The results are shown in FIG. 10. In FIG. 10, the EC 50 The EC (half binding concentration) value reflects the binding capacity of the antibody. 50 The smaller the value, the stronger the binding ability of the antibody. According to the experimental results, as the concentration of the test antibody increases from 0 μg / mL to 20 μg / mL, the parent% value gradually increases until it approaches stability, EC50 It is shown that the value = 0.08012 μg / mL. It can be seen that the test antibody 23E12 has good binding ability with the mouse-TrKA receptor on the surface of HEK293T-mouse-TrKA cells. EXAMPLES

[0095] Detection of the inhibitory effect of test antibodies on the binding of mouse NGF and mouse TrKA by ELISA In the test, IgG4 was used as a negative control (antibody that does not bind to mouse-NGF and mouse-TrkA), and mouse-NGF was biotinylated. Mouse-NGF could bind to mouse-TrkA protein, and anti-mouse-TrkA monoclonal antibody could also bind to mouse-TrkA protein. A competitive experiment was designed to detect the binding of mouse-NGF and mouse-TrkA protein under different concentrations (2.5 μg / mL, 0.25 μg / mL) of test antibodies (23E12, 21E5) by ELISA method. Thus, the inhibitory effect of test antibodies on the binding of mouse-NGF and mouse-TrKA was studied. The experimental results are shown in Figure 11. In Figure 11, the OD 450 The value reflects the mouse-NGF signal binding to mouse-TrkA. The lower the reading, the weaker the mouse-NGF signal binding to mouse-TrkA, and the greater the effect of the antibody on inhibiting the binding of mouse-NGF and mouse-TrKA, as shown in Figure 11, compared with the negative control group, the mouse-NGF signal binding to mouse-TrkA is significantly reduced under the effect of different concentrations (2.5μg / mL, 0.25μg / mL) of the test antibody (23E12, 21E5). It can be seen that the test antibody 23E12 and 21E5 can inhibit the binding of mouse-NGF and mouse-TrKA. EXAMPLES

[0096] Detection of the inhibitory effect of test antibodies on the binding of mouse-NGF and mouse-TrKA by flow cytometry Mouse-NGF was biotinylated. Mouse-NGF could bind to the extracellular domain of mouse-TrkA protein on HEK293T-mouse-TrkA cells, and anti-mouse-TrkA monoclonal antibody could also bind to the extracellular domain of mouse-TrkA protein on HEK293T-mouse-TrkA cells. A competition experiment was designed to detect the binding of mouse-NGF to the extracellular domain of mouse-TrkA protein on HEK293T-mouse-TrkA cells under various concentrations (20μg / mL, 10μg / mL, 5μg / mL, 2.5μg / mL, 1.25μg / mL, 0.625μg / mL, 0.313μg / mL, 0.156μg / mL, 0.078μg / mL, 0.039μg / mL, 0.019μg / mL) of the test antibody by flow cytometry, and thus the inhibitory effect of the test antibody on the binding of mouse-NGF and mouse-TrKA was studied. The experimental results are shown in Figure 12. In Figure 12, the parent % value reflects mouse-NGF binding to the extracellular domain of mouse-TrKA protein on HEK293T-mouse-TrkA cells. The lower the reading, the weaker the mouse-NGF signal binding to the extracellular domain of mouse-TrkA protein on HEK293T-mouse-TrkA cells, and the greater the effect of the antibody on inhibiting the binding of mouse-NGF and mouse-TrKA; as shown in FIG. 12, with increasing test antibody concentration, the parental % value gradually decreases until it approaches zero, i.e., the signal of mouse-NGF binding to the extracellular domain of mouse-TrkA protein gradually decreases until mouse-NGF no longer binds to the extracellular domain of mouse-TrkA protein and all binding of mouse-NGF and mouse-TrkA is inhibited. IC 50 = 0.05147 μg / mL. It can be seen that within a certain concentration range, the test antibody 23E12 can dose-dependently inhibit the binding of mouse-NGF and mouse-TrkA at the cellular level. EXAMPLES

[0097] Detection of the inhibitory effect of test antibodies on the binding of human NGF and human TrKA by ELISA Mouse-NGF was biotinylated. Mouse-NGF could bind to mouse-TrkA protein, and anti-mouse-TrkA monoclonal antibody could also bind to mouse-TrkA protein. A competitive experiment was designed to detect the binding of mouse-NGF and mouse-TrkA protein under various concentrations (20μg / mL, 10μg / mL, 5μg / mL, 2.5μg / mL, 1.25μg / mL, 0.625μg / mL, 0.313μg / mL, 0.156μg / mL, 0.078μg / mL, 0.039μg / mL, 0.019μg / mL) of the test antibody by ELISA, thus studying the inhibitory effect of the test antibody on the binding of mouse-NGF and mouse-TrkA. The experimental results are shown in Figure 13. In Figure 13, the OD 450 The values ​​reflect the mouse-NGF signal binding to mouse-TrkA. The lower the reading, the weaker the mouse-NGF signal binding to mouse-TrkA and the greater the effect of the antibody on inhibiting the binding of mouse-NGF and mouse-TrKA, as the concentration of the test antibody increases, as shown in FIG. 450 The value gradually decreases until it approaches zero, i.e., the signal of mouse-NGF binding to the extracellular domain of mouse-TrkA protein gradually decreases until mouse-NGF hardly binds to the extracellular domain of mouse-TrkA protein, and the binding of mouse-NGF and mouse-TrkA is almost completely inhibited. It can be seen that within a certain concentration range, the test antibodies 23E12, 20C8, 21E5, 27H3, 4H4, 2A5 can dose-dependently inhibit the binding of NGF and TrkA at the molecular level. EXAMPLES

[0098] Evaluation of in vivo analgesic activity of test antibodies using a formalin-induced pain model The formalin inflammatory pain model is a validated pain model that produces continuous pain stimuli and responses, rather than transient pain stimuli and responses, by injecting formalin. This model produces two stages of pain: stage I chemically stimulated pain and stage II inflammatory pain. The pain responses evoked by this model are reproducible and measurable. This model is one of the best models for preclinical pain research and is widely used to evaluate the analgesic effects of various drugs. For the test, 6-8 week old male ICR mice were selected and randomly divided into six groups according to their body weight before modeling: model group (subcutaneous injection of saline), tanezumab 60μg / mouse dose group, 20C8 60μg / mouse dose group, 21E5 60μg / mouse dose group, 23E12 15μg / mouse dose group, and 23E12 60μg / mouse dose group. 10 mice in each group. Drugs were administered subcutaneously. After 18 hours, 15 μL of 2.5% formalin solution was subcutaneously injected into the dorsal side of the right hind leg of the mice. The number of times the mice lifted their paws within 45 minutes was observed. 1-15 minutes is stage I pain, and 16-45 minutes is stage II pain. Generally speaking, stage I pain reflects acute pain, and stage II pain reflects chronic pain. The results are shown in Figure 14 (data in Figure 14 are mean ± SEM, n = 10 / group, compared with the vehicle model group). * p<0.05, using single factor analysis of variance+LSD multiple comparison test). The number of paw lifts in Figure 14 reflects the pain intensity of mice after modeling. The lower the number of paw lifts, the weaker the pain. The results show that compared with the model group, the 23E12 60μg / mouse dose group can significantly reduce the number of paw lifts in mice in stage II pain, and other dose groups cannot reduce the number of paw lifts in mice after modeling. Conclusion: In the formalin-induced pain model, subcutaneous injection of the test antibody 23E12 at 60μg / mouse is effective for chronic pain, while the positive drug tanezumab does not show analgesic effect and may be insensitive to the formalin-induced pain model. EXAMPLES

[0099] Evaluation of the in vivo analgesic activity of test antibodies using a complete Freund's adjuvant-induced inflammatory pain model The Freund's complete adjuvant-induced inflammatory pain model is a pain model that produces chronic inflammatory pain stimuli similar to osteoarthritis and response by injecting Freund's complete adjuvant into the palm of the mouse. Pain is measured by mechanical pain testing. The greater the intensity of mechanical stimulation, the more tolerant the animal is to pain. For the test, 8-week-old male ICR mice were selected and randomly divided into 6 groups (15 mice / group) according to pain sensitivity before modeling: control group (injection of 25 μl saline + subcutaneous injection of saline in the plantar of the left hind paw of the mouse), model group (injection of 25 μl CFA-induced inflammatory pain + subcutaneous injection of saline in the left hind paw of the mouse), tanezumab 60 μg / mouse dose group, 23E12 60 μg / mouse dose group. Four days after modeling, the drug was subcutaneously injected, and 36 hours after administration, mechanical pain testing was performed. The results are shown in Figure 15 (data in Figure 15 are mean ± SEM, n = 15 / group, compared to the blank control group). # p □ 0.05, ## p □ 0.01, compared with the model group; * p □ 0.05, ** p □ 0.01, using non-parametric statistical analysis + independent sample t-test). The vertical axis represents the intensity of mechanical stimulation. The higher the pressure threshold of mouse paw withdrawal, the better the analgesic effect. The results show that compared with the model group, subcutaneous injection of 60 μg / mouse 23E12 and 60 μg / mouse tanezumab can significantly increase the pressure threshold of mouse paw withdrawal (P<0.01, P<0.05), exhibiting analgesic effect. Both analgesic effects are comparable. Conclusion: Subcutaneous injection of 60 μg / mouse test antibody 23E12 exhibits analgesic effect in complete Freund's adjuvant-induced inflammatory pain model. EXAMPLES

[0100] Toxicity testing of test antibodies In this example, the toxicity tests to be performed by the inventors are shown in Table 2 below.

[0101] [Table 2A]

[0102] [Table 2B]

[0103] Among them, the above experimental process and experimental conclusions are as follows: Toxicity Testing: Single-dose toxicity SD rats and cynomolgus monkeys were observed for target organ toxicity. Relevant symptoms such as effects on body weight, food intake, ophthalmoscopy, electrocardiogram, hematology, clinical biochemistry, urine and organ weights were observed. Maximum tolerated dose (MTD) and sufficient safety margins were provided. Results showed that no animal deaths were observed and diffuse inflammatory infiltrates were observed in organs and tissues, but target organ toxicity was not indicated by clinical pathology or histological analysis. No significant effects were observed on body weight, food intake, ophthalmoscopy, electrocardiogram, hematology, clinical biochemistry, urine and organ weights. No significant abnormalities were observed in gross anatomy.

[0104] Repeated dose toxicity: SD rats and cynomolgus monkeys were injected intravenously with the drug once a week for 4 consecutive weeks. Clinical pathology and histology were observed to determine whether a target organ toxicity NOAEL was observed. Effects on body weight, food intake, ophthalmoscopy, electrocardiogram, hematology, clinical biochemistry, urine and organ weights were observed. Safety windows were calculated by immunogenicity (ADA (anti-drug antibody) assay), immunotoxicity studies (assessments included hematological differential counts of leukocytes (including macrophages), clinical chemistry (globulins and albumin:globulin ratios)), organ masses (thymus, spleen, lymph nodes) and histopathology (lymphoid organs and tissues). Results showed that no animals died and diffuse inflammatory infiltrates were observed in organs and tissues, but no target organ toxicity was indicated regardless of clinical pathology or histological analysis. No nivolumab-related symptoms were observed and there were no significant effects on body weight, food intake, ophthalmoscopy, electrocardiogram, hematology, clinical biochemistry, urinalysis, or organ weights. Gross anatomy revealed no significant abnormalities.

[0105] General safety pharmacology: SD rats were intravenously injected with a single dose of the drug. The effects of the drug on the central nervous system, cardiovascular system and respiratory system were observed. The results showed that the drug had no effect on the central nervous system, cardiovascular system and respiratory system.

[0106] Local administration safety test (vascular stimulation test and in vitro hemolysis test): The drug was intravenously injected into New Zealand white rabbits to test whether the drug had any effect on blood vessels and blood. The results showed that the drug had no effect on blood vessels and blood.

[0107] High sensitization test (systemic active allergy test in guinea pigs): The drug was injected intravenously into guinea pigs in a single dose to observe whether the drug has sensitization. The results showed that the drug has no sensitization.

[0108] Throughout this specification, references to "an embodiment," "some embodiments," "one embodiment," "another embodiment," "an example," "particular example," or "some examples" mean that a particular feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the present disclosure. Thus, the appearance of phrases such as "in some embodiments," "in one embodiment," "in an embodiment," "in another example," "in an example," "in a specific example," or "in some examples" in various places throughout this specification do not necessarily refer to the same embodiment or example of the present disclosure. Furthermore, particular features, structures, materials, or characteristics may be combined in any suitable manner in one or more embodiments or examples. Furthermore, without any inconsistency, one of ordinary skill in the art may combine the features of the various embodiments or examples described herein with the various embodiments or examples.

[0109] While illustrative embodiments have been shown and described, it will be understood by those skilled in the art that the above embodiments cannot be construed as limiting the disclosure, and that changes, substitutions, and modifications can be made in the embodiments without departing from the spirit, principles, and scope of the disclosure.

Claims

1. An antibody or an antigen-binding fragment thereof capable of specifically recognizing TrkA, the antibody comprising: Heavy chain variable region CDR1, CDR2, CDR3 sequences shown in SEQ ID NO: 4, SEQ ID NO: 5, and SEQ ID NO: 6, and Light chain variable region CDR1, CDR2, and CDR3 sequences shown in SEQ ID NO: 31, SEQ ID NO: 32, and SEQ ID NO: 33 An antibody or antigen-binding fragment thereof comprising the CDR sequences of:

2. The antibody or antigen-binding fragment thereof according to claim 1, wherein the antibody has a heavy chain variable region of the amino acid sequence shown in SEQ ID NO:

56.

3. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody has a light chain variable region of the amino acid sequence shown in SEQ ID NO:

65.

4. the heavy chain constant region of the antibody is derived from human IgG4; The antibody or antigen-binding fragment thereof of claim 1, wherein the Fc region of the antibody has S10P, F16A, L17A, R191K mutations and 229K deletion mutations compared to the wild type.

5. The antibody or antigen-binding fragment thereof of claim 4, wherein the full-length sequence of the constant region of the antibody is set forth in SEQ ID NO:

74.

6. The antibody or antigen-binding fragment thereof described in claim 1, wherein the light chain of the antibody has a constant region, and the full-length sequence of the constant region is set forth in SEQ ID NO:

75.

7. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody has a heavy chain having the amino acid sequence set forth in SEQ ID NO: 77 and a light chain having the amino acid sequence set forth in SEQ ID NO:

86.

8. The antibody or antigen-binding fragment thereof of claim 1, wherein the antibody is a single chain antibody fragment, a multimeric antibody, a CDR-grafted antibody, or a small molecule antibody.

9. 9. The antibody or antigen-binding fragment thereof of claim 8, wherein the antibody is a single-chain antibody fragment and the antibody has an amino acid sequence as set forth in SEQ ID NO: 96 or 97.

10. Encoding the antibody or antigen-binding fragment thereof of any one of claims 1 to 9, DNA, A nucleic acid molecule having the nucleotide sequence set forth in SEQ ID NO:113, or the nucleotide sequence set forth in SEQ ID NO:122, or the nucleotide sequence set forth in SEQ ID NO:132 or 133.

11. A pharmaceutical composition comprising an antibody according to any one of claims 1 to 9 or a nucleic acid molecule according to claim 10.

12. A kit for detecting TrkA comprising an antibody according to any one of claims 1 to 9.

13. An antibody according to any one of claims 1 to 9, a nucleic acid molecule according to claim 10 or a pharmaceutical composition according to claim 11 for use in the preparation of a medicament for the treatment or prevention of pain, cancer, inflammation or inflammatory diseases, neurodegenerative diseases, Sjogren's syndrome, endometriosis, diabetic peripheral neuropathy, prostatitis, pelvic pain syndrome, diseases associated with imbalance in the regulation of bone remodelling and diseases caused by abnormal signalling of connective tissue growth factor.

14. 14. The antibody, nucleic acid molecule, or pharmaceutical composition of claim 13, wherein the medicament is used to treat or prevent neuropathic pain, inflammatory pain, cancer-related pain, fracture-related pain, surgery-related pain, inflammatory lung disease, interstitial cystitis, painful bladder syndrome, inflammatory bowel disease, inflammatory skin disease, Raynaud's syndrome, idiopathic pulmonary fibrosis, scarring (hypertrophic, keloid type and other forms), sclerosis, endomyocardial fibrosis, atrial fibrosis, myelofibrosis, progressive massive fibrosis (lung), nephrogenic systemic fibrosis, scleroderma, systemic sclerosis, arthrofibrosis, ocular fibrosis, non-small cell lung cancer, papillary thyroid carcinoma, glioblastoma multiforme, colorectal cancer, melanoma, cholangiocarcinoma or sarcoma, acute myeloid leukemia, large cell neuroendocrine carcinoma, neuroblastoma, prostate cancer, pancreatic cancer, melanoma, head and neck squamous cell carcinoma, or gastric cancer.

15. 11. An antibody according to any one of claims 1 to 9 or a nucleic acid molecule according to claim 10 for use in the preparation of a kit for detecting TrkA or for diagnosing a TrkA-related disease.

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