Antibody drug conjugates targeting receptor tyrosine kinase-like orphan receptor 1

Abstract

The application belongs to the field of biological medicine and provides an antibody drug conjugate targeting receptor tyrosine kinase-like orphan receptor 1, wherein the antibody drug conjugate comprises an antibody, a linker and a cytotoxic drug connected in sequence. The application also provides a pharmaceutical composition comprising the antibody drug conjugate and uses thereof. The provided antibody drug conjugate achieves excellent antitumor activity or / and better safety. The provided antibody drug conjugate can be used for the treatment of tumors.

Description

[0001] Citation of relevant applications This disclosure claims priority to Chinese Patent Application No. 202510061746.1, filed on January 14, 2025, the entire contents of which are incorporated herein by reference and used for all purposes. Technical Field

[0002] This disclosure pertains to the biomedical field and relates to antibody-drug conjugates targeting the receptor tyrosine kinase-like orphan receptor 1 (ROR1), comprising an associated antibody or antigen-binding fragment thereof, a linker, and a cytotoxic drug. This disclosure also relates to the use and methods of said antibody-drug conjugates in treating diseases expressing ROR1, such as tumors. Background Technology

[0003] Receptor tyrosine kinase-like orphan receptor 1 (ROR1) belongs to the ROR receptor family, a family of cell tight junction proteins. This family consists of two closely related type I transmembrane proteins, ROR1 and ROR2, which participate in intercellular signaling, intracellular signal transduction, and regulate cell proliferation, differentiation, and metastasis. The structure of ROR1 includes an extracellular region, a transmembrane region, and an intracellular region. The extracellular region consists of an Ig-like domain, a cysteine-rich coiled domain, and a kringle domain, while the intracellular region consists of a tyrosine kinase region, a serine / threonine-rich region, and a proline-rich region.

[0004] ROR1 is selectively overexpressed in many solid and hematologic malignancies, such as leukemia, breast cancer, ovarian cancer, and lung adenocarcinoma, but is not significantly expressed in normal adult tissues. Multiple studies have shown that ROR1 can promote tumor growth in vitro and in vivo, playing a promoting role in the malignant progression of tumors, making ROR1 a potential target for targeted cancer therapy.

[0005] Antibody-drug conjugates (ADCs) are a class of drugs that combine the high specificity and targeting of therapeutic antibodies with the high cytotoxic activity of cytotoxic drugs, with the therapeutic antibody portion and the cytotoxic drug portion linked by an intermediate linker. While there are existing reports on the development of antibody-drug conjugates for anti-tumor research targeting the ROR1 target, these are limited, and more available options are still needed. Summary of the Invention

[0006] Antibody-drug conjugates (ADCs) This disclosure provides an antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, comprising an anti-ROR1 antibody or its antigen-binding fragment and a cytotoxic drug, wherein the anti-ROR1 antibody or its antigen-binding fragment is conjugated to the cytotoxic drug.

[0007] This disclosure provides an antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, comprising the anti-ROR1 antibody of this disclosure or its antigen-binding fragment.

[0008] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment is a murine antibody, a chimeric antibody, a humanized antibody, or a human antibody. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment is a monoclonal antibody, a monospecific antibody, a multispecific antibody, a Fab fragment, an F(ab')2 fragment, an Fd fragment, an Fv fragment, a dAb, isolated CDR, scFv, scFab, a nanobody, or a combination thereof.

[0009] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain CDR (HCDR)1 comprising the amino acid sequence shown in SEQ ID NO: 1, 9, 17 or 25; an HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 2, 10, 18, 26, 49 or 50; an HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 3, 11, 19 or 27; a light chain CDR (LCDR)1 comprising the amino acid sequence shown in SEQ ID NO: 4, 12, 20 or 28; an LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 5, 13, 21 or 29; and an LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 6, 14, 22 or 30.

[0010] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 1, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 2, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 3, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 4, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 5, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 6.

[0011] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 50, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14; X1 in SEQ ID NO: 50 is selected from S or A, X2 is selected from A or Q, and X3 is selected from K or Q. In some specific embodiments, X1 in SEQ ID NO: 50 is S, X2 is A, and X3 is K. In some specific embodiments, X1 in SEQ ID NO: 50 is A, X2 is Q, and X3 is Q.

[0012] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1 containing the amino acid sequence shown in SEQ ID NO: 9, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 10, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 11, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 12, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 14.

[0013] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1 containing the amino acid sequence shown in SEQ ID NO: 9, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 49, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 11, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 12, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 14.

[0014] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1 containing the amino acid sequence shown in SEQ ID NO: 17, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 18, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 19, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 20, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 21, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 22.

[0015] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1 containing the amino acid sequence shown in SEQ ID NO: 25, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 26, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 27, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 28, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 30.

[0016] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 7, 15, 23, 31, 51, 52, 56, 57, 58, 60, or 61, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 8, 16, 24, 32, 53, 54, 55, 59, 62, 63, or 64. The CDRs are defined, for example, according to Kabat, IMGT, Chothia, Contact, AbM, and / or CCG.

[0017] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 7, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 8.

[0018] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 15, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 16.

[0019] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 23, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 24.

[0020] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2 and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 31, and LCDR1, LCDR2 and LCDR3 in the light chain variable region as shown in SEQ ID NO: 32.

[0021] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2 and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 51, and LCDR1, LCDR2 and LCDR3 in the light chain variable region as shown in SEQ ID NO: 53.

[0022] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 51, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 54.

[0023] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 51, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 55.

[0024] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 52, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 53.

[0025] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 52, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 54.

[0026] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 52, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 55.

[0027] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 56, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 59.

[0028] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 57, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 59.

[0029] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 58, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 59.

[0030] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 60, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 62.

[0031] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 60, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 63.

[0032] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 60, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 64.

[0033] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 61, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 62.

[0034] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 61, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 63.

[0035] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 61, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 64.

[0036] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequences shown in SEQ ID NO: 7, 15, 23, 31, 51, 52, 95%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequences shown in SEQ ID NO: 8, 16, 24, 32, 53, 54, 55, 59, 62, 63, or 64. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 7, 15, 23, 31, 51, 52, 95%, 96%, 97%, 98%, 99%, or 100%; and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 8, 16, 24, 32, 53, 54, 55, 59, 62, 63, or 64.

[0037] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 7. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 8. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 7, and a light chain variable region having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 8. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 containing the amino acid sequence shown in SEQ ID NO: 1, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 2, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 3, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 4, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 5, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 6.

[0038] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 15. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 16. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 15, and a light chain variable region having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 16. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 10, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0039] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 23. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 24. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 23, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 24. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 17, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 18, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 19, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 20, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 21, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 22.

[0040] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 32. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 32. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 31, and a light chain variable region having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 32. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0041] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 51. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 53. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 53. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 49, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0042] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 51. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 54. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 54. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 49, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0043] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 51. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 55. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 55. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 49, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0044] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 52. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 53. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 52, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 53. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 10, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0045] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 52. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 54. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 52, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 54. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 10, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0046] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 52. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 55. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 52, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 55. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 10, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0047] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 56. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 59. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 56, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 59. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 17, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 18, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 19, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 20, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 21, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 22.

[0048] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 57. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 59. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 57, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 59. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 17, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 18, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 19, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 20, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 21, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 22.

[0049] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 58. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 59. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 58, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 59. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 17, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 18, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 19, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 20, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 21, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 22.

[0050] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 60. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 62. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 60, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 62. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0051] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 60. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 63. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 60, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 63. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0052] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 60. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 64. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 60, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 64. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0053] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 62. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 62. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 62. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0054] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 63. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 63. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 63. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0055] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 61. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 64. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 64. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0056] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 7, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 8.

[0057] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 15, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 16.

[0058] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 23, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 24.

[0059] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 31, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 32.

[0060] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 53.

[0061] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 54.

[0062] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 55.

[0063] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 52, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 53.

[0064] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 52, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 54.

[0065] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 52, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 55.

[0066] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 56, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 59.

[0067] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 57, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 59.

[0068] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 58, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 59.

[0069] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 60, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 62.

[0070] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 60, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 63.

[0071] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 60, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 64.

[0072] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 62.

[0073] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 63.

[0074] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 64.

[0075] In this disclosure, the antigen-binding fragment can be a Fab fragment, an F(ab')2 fragment, an Fd fragment, an Fv fragment, dAb, scFv, a single-chain Fab (scFab), or a separated CDR region. In this disclosure, the anti-ROR1 antibody or its antigen-binding fragment can be murine, chimeric, or humanized. Humanization can reduce immunogenicity. Therefore, in some embodiments, the anti-ROR1 antibody or its antigen-binding fragment is humanized.

[0076] Tables S1 and S2 provide some exemplary anti-ROR1 antibodies or their antigen-binding fragments (e.g., chimeric antibodies ch3G9B1, ch29H5D8, ch135A2B10, ch137G11D10 and humanized antibodies hz29H5D8-1.1, hz29H5D8-1.2, hz29H5D8-2.1, hz29H5D8-2.2, hz29H5D8-3.1, hz29H5D8-3). 2. The amino acid sequence information of the CDR and variable regions of hz135A2B10-1.1, hz135A2B10-1.2, hz135A2B10-1.3, hz137G11D10-1.1, hz137G11D10-1.2, hz137G11D10-2.1, hz137G11D10-2.2, hz137G11D10-3.1, and hz137G11D10-3.2.

[0077] Table S1. Amino acid sequences of CDR and variable regions (SEQ ID NO:)

[0078] For the variable region amino acid sequence of an antibody or its antigen-binding fragment, those skilled in the art can determine which residues constitute a specific CDR using conventional methods. As is known to those skilled in the art, the CDR of an antibody can be defined in various ways, such as using numbering systems / methods such as Kabat, Chothia, IMGT, AbM, CCG, or Contact; or using a combination of two or more of these numbering systems / methods; or using a combined numbering system that incorporates both Kabat and Chothia, which combines the ranges defined by Kabat and Chothia to form a larger range. The precise numbering and location of the CDR differ in different numbering systems. Those skilled in the art should understand that, unless otherwise specified, the terms "CDR" and "complementarity-determining region" for a given antibody or its region (e.g., variable region) should be understood to encompass the complementarity-determining region defined by any known scheme. Although the CDRs claimed in this disclosure are based on the sequences shown in Table S1 (one definition), the amino acid sequences corresponding to other CDR definition rules should also fall within the scope of protection of this disclosure.

[0079] The VH and / or VL sequences (or CDR sequences) of other antibodies that bind to ROR1 can be "mixed and paired" with the VH and / or VL sequences (or CDR sequences) of the anti-ROR1 antibody or its antigen-binding fragment. Preferably, when the VH and VL chains (or their CDRs) are mixed and paired, the VH sequence in a particular VH / VL pair can be replaced by a structurally similar VH sequence. Similarly, it is preferable to replace the VL sequence in a particular VH / VL pair with a structurally similar VL sequence.

[0080] Therefore, in one embodiment, the anti-ROR1 antibody or its antigen-binding fragment comprises: (a) The heavy chain variable region, which contains the VH amino acid sequence listed in Table S1; and (b) A light chain variable region comprising the VL amino acid sequence listed in Table S1, or the VL of another anti-ROR1 antibody or its antigen-binding fragment thereof, wherein the antibody or its antigen-binding fragment specifically binds to ROR1.

[0081] In another embodiment, the anti-ROR1 antibody or its antigen-binding fragment comprises: (a) HCDR1, HCDR2 and HCDR3 listed in Table S1; and (b) LCDR1, LCDR2 and LCDR3 listed in Table S1, or another anti-ROR1 antibody or its antigen-binding fragment thereof, wherein the antibody or its antigen-binding fragment specifically binds to ROR1.

[0082] In another embodiment, the anti-ROR1 antibody or its antigen-binding fragment includes HCDR2 listed in Table S1 and other CDR(s) of anti-ROR1 antibodies or their antigen-binding fragments, such as HCDR1 and / or HCDR3 of other anti-ROR1 antibodies or their antigen-binding fragments, and / or LCDR1, LCDR2 and / or LCDR3 of other anti-ROR1 antibodies or their antigen-binding fragments.

[0083] In another embodiment, the anti-ROR1 antibody or its antigen-binding fragment comprises HCDR2 listed in Table S1, and HCDR3 and / or LCDR3 listed in Table S1, as well as CDR(s) of other anti-ROR1 antibodies or their antigen-binding fragments, wherein the antibody or its antigen-binding fragment specifically binds to ROR1. In another embodiment, the anti-ROR1 antibody or its antigen-binding fragment further comprises LCDR2 listed in Table S1, as well as CDR(s) of other anti-ROR1 antibodies, wherein the antibody or its antigen-binding fragment specifically binds to ROR1. In another embodiment, the anti-ROR1 antibody or its antigen-binding fragment further comprises HCDR1 or LCDR1 listed in Table S1, as well as CDR(s) of other anti-ROR1 antibodies, wherein the antibody or its antigen-binding fragment specifically binds to ROR1. These antibodies are preferably (a) competitively binding to ROR1 with the exemplary anti-ROR1 antibodies provided in Tables S1 and S2; (b) retaining the same functional characteristics; (c) binding to the same epitopes; and / or (d) having similar binding affinity.

[0084] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment may further comprise a constant region of an immunoglobulin, or a fragment, analog, variant, or derivative of said constant region. In some embodiments, the constant region comprises a heavy chain constant region and a light chain constant region. In some embodiments, the heavy chain constant region is derived from the heavy chain of human immunoglobulins, such as IgG1, IgG2, IgG3, IgG4, or other classes of immunoglobulins. In some embodiments, the light chain constant region is derived from the light chain of human immunoglobulins, such as the κ or λ light chain of human immunoglobulins. In some embodiments, the anti-ROR1 antibody is an IgG1, IgG2, IgG3, or IgG4 isotype. In some embodiments, the constant region may comprise any modification disclosed herein or known in the art, such as the insertion, deletion, substitution, or chemical modification of amino acids. The C-terminal lysine of the heavy chain constant region may be present or absent. In some embodiments, the constant region comprises a mutation that alters effector function. In some embodiments, any amino acid residue of the constant region may be substituted with any allotype amino acid residue.

[0085] In some specific embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 33, 37, 41, 47, 69, 71, 73, 81, 87, 89, or 91. In some specific embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 35, 39, 43, 47, 69, 71, 73, 81, 87, 89, or 91. In some specific embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 33, 37, 41, 47, 69, 71, 73, 81, 87, 89, or 91, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 35, 39, 43, 47, 69, 71, 73, 81, 87, 89, or 91.

[0086] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 33. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 35. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 33, and a light chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 35. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 1, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 2, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 3, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 4, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 5, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 6.

[0087] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 37. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 39. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 37, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 39. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 10, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0088] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 41. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 43. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 41, and a light chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 43. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 17, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 18, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 19, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 20, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 21, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 22.

[0089] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 45. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 47. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 45, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 47. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0090] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 65. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 69. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 65, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 69. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 49, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0091] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 65. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 71. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 65, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 71. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 49, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0092] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 65. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 73. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 65, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 73. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 49, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0093] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 67. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 69. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 67, and a light chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 69. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 10, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0094] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 67. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 71. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 67, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 71. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 10, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0095] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 67. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 73. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 67, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 73. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 10, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14.

[0096] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 75. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 81. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 75, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 81. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 17, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 18, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 19, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 20, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 21, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 22.

[0097] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 77. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 81. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 77, and a light chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 81. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 17, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 18, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 19, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 20, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 21, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 22.

[0098] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 79. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 81. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 79, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 81. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 17, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 18, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 19, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 20, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 21, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 22.

[0099] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 83. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 87. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 83, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 87. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0100] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 83. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 89. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 83, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 89. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0101] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 83. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 91. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 83, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 91. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0102] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 85. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 87. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 85, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 87. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0103] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 85. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 89. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 85, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 89. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0104] In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 85. In some embodiments, the anti-ROR1 antibody comprises a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 91. In some embodiments, the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 85, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 91. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0105] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 33, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 35.

[0106] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 37, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 39.

[0107] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 41, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 43.

[0108] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 45, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 47.

[0109] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 65, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 69.

[0110] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 65, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 71.

[0111] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 65, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 73.

[0112] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 67, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 69.

[0113] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 67, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 71.

[0114] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 67, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 73.

[0115] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 75, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 81.

[0116] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 77, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 81.

[0117] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 79, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 81.

[0118] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 83, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 87.

[0119] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 83, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 89.

[0120] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 83, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 91.

[0121] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 85, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 87.

[0122] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 85, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 89.

[0123] In some embodiments, the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 85, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 91.

[0124] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 33 and a light chain of the amino acid sequence shown in SEQ ID NO: 35.

[0125] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 37 and a light chain of the amino acid sequence shown in SEQ ID NO: 39.

[0126] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 41 and a light chain of the amino acid sequence shown in SEQ ID NO: 43.

[0127] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 45 and a light chain of the amino acid sequence shown in SEQ ID NO: 47.

[0128] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 65 and a light chain of the amino acid sequence shown in SEQ ID NO: 69.

[0129] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 65 and a light chain of the amino acid sequence shown in SEQ ID NO: 71.

[0130] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 65 and a light chain of the amino acid sequence shown in SEQ ID NO: 73.

[0131] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 67 and a light chain of the amino acid sequence shown in SEQ ID NO: 69.

[0132] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 67 and a light chain of the amino acid sequence shown in SEQ ID NO: 71.

[0133] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 67 and a light chain of the amino acid sequence shown in SEQ ID NO: 73.

[0134] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 75 and a light chain of the amino acid sequence shown in SEQ ID NO: 81.

[0135] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 77 and a light chain of the amino acid sequence shown in SEQ ID NO: 81.

[0136] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 79 and a light chain of the amino acid sequence shown in SEQ ID NO: 81.

[0137] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 83 and a light chain of the amino acid sequence shown in SEQ ID NO: 87.

[0138] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 83 and a light chain of the amino acid sequence shown in SEQ ID NO: 89.

[0139] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 83 and a light chain of the amino acid sequence shown in SEQ ID NO: 91.

[0140] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 85 and a light chain of the amino acid sequence shown in SEQ ID NO: 87.

[0141] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 85 and a light chain of the amino acid sequence shown in SEQ ID NO: 89.

[0142] In some embodiments, the anti-ROR1 antibody comprises a heavy chain of the amino acid sequence shown in SEQ ID NO: 85 and a light chain of the amino acid sequence shown in SEQ ID NO: 91.

[0143] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 33, and the amino acid sequence of the light chain is shown in SEQ ID NO: 35.

[0144] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 37, and the amino acid sequence of the light chain is shown in SEQ ID NO: 39.

[0145] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 41, and the amino acid sequence of the light chain is shown in SEQ ID NO: 43.

[0146] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 45, and the amino acid sequence of the light chain is shown in SEQ ID NO: 47.

[0147] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 65, and the amino acid sequence of the light chain is shown in SEQ ID NO: 69.

[0148] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 65, and the amino acid sequence of the light chain is shown in SEQ ID NO: 71.

[0149] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 65, and the amino acid sequence of the light chain is shown in SEQ ID NO: 73.

[0150] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 67, and the amino acid sequence of the light chain is shown in SEQ ID NO: 69.

[0151] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 67, and the amino acid sequence of the light chain is shown in SEQ ID NO: 71.

[0152] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 67, and the amino acid sequence of the light chain is shown in SEQ ID NO: 73.

[0153] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 75, and the amino acid sequence of the light chain is shown in SEQ ID NO: 81.

[0154] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 77, and the amino acid sequence of the light chain is shown in SEQ ID NO: 81.

[0155] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 79, and the amino acid sequence of the light chain is shown in SEQ ID NO: 81.

[0156] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 83, and the amino acid sequence of the light chain is shown in SEQ ID NO: 87.

[0157] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 83, and the amino acid sequence of the light chain is shown in SEQ ID NO: 89.

[0158] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 83, and the amino acid sequence of the light chain is shown in SEQ ID NO: 91.

[0159] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 85, and the amino acid sequence of the light chain is shown in SEQ ID NO: 87.

[0160] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 85, and the amino acid sequence of the light chain is shown in SEQ ID NO: 89.

[0161] In some embodiments, the amino acid sequence of the heavy chain of the anti-ROR1 antibody is shown in SEQ ID NO: 85, and the amino acid sequence of the light chain is shown in SEQ ID NO: 91.

[0162] Tables S1 and S2 provide CDR, variable region, and full-length amino acid sequence information for some exemplary anti-ROR1 antibodies or their antigen-binding fragments. Some anti-ROR1 antibodies or their antigen-binding fragments have the same CDR, some have the same VH or VL, or have the same heavy or light chain. These exemplary anti-ROR1 antibodies or their antigen-binding fragments can be used to construct the antibody-drug conjugates disclosed herein.

[0163] In some specific embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure, wherein the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1 of the amino acid sequence shown in SEQ ID NO: 25, HCDR2 of the amino acid sequence shown in SEQ ID NO: 26, HCDR3 of the amino acid sequence shown in SEQ ID NO: 27, LCDR1 of the amino acid sequence shown in SEQ ID NO: 28, LCDR2 of the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 of the amino acid sequence shown in SEQ ID NO: 30.

[0164] In some specific embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts provided in this disclosure, wherein the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 60, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 63.

[0165] In some specific embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts provided in this disclosure, wherein the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 60, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 63. Furthermore, in some such embodiments, the anti-ROR1 antibody or its antigen-binding fragment further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0166] In some specific embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure, wherein the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 60, and a light chain variable region of the amino acid sequence shown in SEQ ID NO: 63.

[0167] In some specific embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts provided in this disclosure, wherein the anti-ROR1 antibody comprises: a heavy chain having an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 83, and a light chain having an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 89. Furthermore, in some such embodiments, the anti-ROR1 antibody further comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 25, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 26, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 27, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 28, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 30.

[0168] In some specific embodiments, the antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof provided in this disclosure, wherein the anti-ROR1 antibody comprises: a heavy chain of the amino acid sequence shown in SEQ ID NO: 83, and a light chain of the amino acid sequence shown in SEQ ID NO: 89.

[0169] In some embodiments, the anti-ROR1 antibody comprises two heavy chains and two light chains, or is composed of two heavy chains and two light chains, the heavy chains and light chains being interconnected by disulfide bonds, wherein each heavy chain comprises the aforementioned heavy chain variable region (VH) and heavy chain constant region, wherein the heavy chain variable region (VH) comprises a backbone region (FR) and the aforementioned heavy chain complementarity-determining region (HCDR), and each light chain comprises the aforementioned light chain variable region (VL) and light chain constant region, wherein the light chain variable region (VL) comprises a FR and the aforementioned light chain complementarity-determining region (LCDR), wherein the C-terminus of the heavy chain variable region is connected to the N-terminus of the heavy chain constant region, and the C-terminus of the light chain variable region is connected to the N-terminus of the light chain constant region.

[0170] In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment can be modified, for example, by including the insertion, deletion, or substitution of one or more amino acids. In this document, the modified anti-ROR1 antibody or its antigen-binding fragment retains its activity of specifically binding to ROR1 (e.g., human ROR1, monkey ROR1).

[0171] In the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts provided in this disclosure, a cytotoxic drug is conjugated to an anti-ROR1 antibody or its antigen-binding fragment via a linker. In some specific embodiments, each cytotoxic drug in the antibody-drug conjugates, their stereoisomers, or their pharmaceutically acceptable salts provided in this disclosure is conjugated to an anti-ROR1 antibody or its antigen-binding fragment via a linker. The linkers of this disclosure can be linked to the anti-ROR1 antibody or its antigen-binding fragment by any method known in the art. In some preferred embodiments, the linker is linked to the anti-ROR1 antibody or its antigen-binding fragment via a thiol group and / or an amino group. In some more preferred embodiments, the linker is linked to the anti-ROR1 antibody or its antigen-binding fragment via a thiol group.

[0172] In some embodiments, the linker may be a cleavable linker or a non-cleavable linker. In some embodiments, the linker is a cleavable linker, such as one that is degraded at low pH (including hydrazone bonds, carbonate bonds, etc.), proteolytic (including peptide bonds), or degraded at high glutathione concentrations (including disulfide bonds). Cleavable linkers can cleave within target cells, thereby releasing cytotoxic drugs. In other embodiments, the linker is a non-cleavable linker, such as one containing maleiminohexanoyl. Non-cleavable linkers can release drugs through antibody degradation.

[0173] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts provided in this disclosure, wherein an anti-ROR1 antibody or its antigen-binding fragment is conjugated to one or more cytotoxic drugs, such as alkaloids, antimetabolites, antitumor antibiotics, alkylating agents, and platinum-based drugs. In some embodiments, the cytotoxic drug is a tubulin inhibitor. In some embodiments, the tubulin inhibitor includes, but is not limited to, maytansine, auristatin, dolatatin, tubulolysins, cryptomycins, and eribulin or derivatives thereof. In some specific embodiments, the cytotoxic drug is eribulin or a derivative thereof.

[0174] In some implementations, the cytotoxic drug is linked to the adapter via a functional group, and the antibody-drug conjugate is hydrolyzed within tumor cells to form a free cytotoxic drug, thereby exerting an anti-tumor effect.

[0175] In some implementations, the cytotoxic drug is linked to the adapter via functional groups, and the antibody-drug conjugate degrades within tumor cells to form a free cytotoxic drug, thereby exerting an anti-tumor effect.

[0176] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure are formed by linking a drug-linker of the structure shown in Formula Ia to an anti-ROR1 antibody or its antigen-binding fragment, wherein the drug-linker is attached to the anti-ROR1 antibody or its antigen-binding fragment at the position indicated by * in the structure shown in Formula Ia. Ia; Among them, R 1 Selected from hydrogen atoms, deuterium atoms, and optionally substituted C atoms 1-6 Alkyl groups, optionally substituted C 3-7 cycloalkyl, optionally substituted 3- to 7-membered heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted 5 to 12 heteroaryl groups, R 2 Selected from hydrogen atoms, deuterium atoms, and optionally substituted C atoms 1-6 Alkyl groups, optionally substituted C 3-7 cycloalkyl, optionally substituted 3- to 7-membered heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted 5 to 12 heteroaryl groups; or, R 1 and R 2 The atoms connected thereto form optionally substituted 5- to 8-membered heterocyclic groups. In some embodiments, the drug-connector is attached at the position indicated by * in the structure shown in Formula Ia to the disulfide bond site of the anti-ROR1 antibody or its antigen-binding fragment, preferably via a thioether bond.

[0177] In some implementations, the R 1 and R 2 Each is independently selected from hydrogen atoms or C atoms. 1-5 Alkyl (preferably C) 1-4 Alkyl, such as C 1-3 Alkyl group). In some embodiments, the R... 1 and R 2 Each is independently selected from hydrogen atoms, methyl, ethyl, propyl, or isopropyl. In some embodiments, the R... 1 and R 2 All are hydrogen atoms.

[0178] In some embodiments, the antibody-drug conjugate is formed by linking a drug-connector of the structure shown in Formula Ia-1 to an anti-ROR1 antibody or its antigen-binding fragment, wherein the drug-connector is attached to the anti-ROR1 antibody or its antigen-binding fragment at the position indicated by * in the structure shown in Formula Ia-1. Ia-1.

[0179] In some embodiments, the drug-connector is attached to the disulfide bond site of the anti-ROR1 antibody or its antigen-binding fragment at the position indicated by * in the structure shown in Formula Ia, preferably via a thioether bond.

[0180] The number of cytotoxic drugs linked to the anti-ROR1 antibody or its antigen-binding fragment in the antibody-drug conjugate, its stereoisomer, or its pharmaceutically acceptable salt provided in this disclosure can vary, such that the antibody-drug conjugate (ADC) can be heterogeneous. That is, the antibody-drug conjugate includes an anti-ROR1 antibody or its antigen-binding fragment linked with different numbers of cytotoxic drugs. For example, one molecule of anti-ROR1 antibody or its antigen-binding fragment may be linked with 0 (i.e., without cytotoxic drugs), 1, 2, 3, 4, 5, 6, 7, 8, or more molecules of cytotoxic drugs.

[0181] By controlling the proportions of antibodies or their antigen-binding fragments linked to different numbers of cytotoxic drugs, antibody-drug conjugates, their stereoisomers, or their pharmaceutically acceptable salts with different drug-antibody ratios (DARs) can be generated. It should be understood that the DAR is the average number of cytotoxic drugs linked to each antibody or its antigen-binding fragment in the antibody-drug conjugate, its stereoisomers, or their pharmaceutically acceptable salts. For example, "DAR of 4" means that such an antibody-drug conjugate, its stereoisomers, or their pharmaceutically acceptable salts comprises a heterogeneous mixture in which each antibody or its antigen-binding fragment is linked to the same or different numbers of cytotoxic drugs (e.g., each antibody or its antigen-binding fragment is linked to 0, 1, 2, 3, 4, 5, 6, 7, and / or 8 cytotoxic drugs), but the average number of cytotoxic drugs linked to each antibody or its antigen-binding fragment is 4. Similarly, "DAR of 8" means that the average number of cytotoxic drugs linked to each antibody or its antigen-binding fragment in the antibody-drug conjugate, its stereoisomers, or their pharmaceutically acceptable salts is 8. Since DAR represents the average number of connections, DAR can be an integer or a decimal.

[0182] In some of the above embodiments, the DAR of the antibody-drug conjugate is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8. In some of the above embodiments, the DAR of the antibody-drug conjugate is 3.5-5. In some of the above embodiments, the DAR of the antibody-drug conjugate is 3.5-4.5. In some of the above embodiments, the DAR of the antibody-drug conjugate is 7-8. In some of the above embodiments, the DAR of the antibody-drug conjugate is 7.5-8. In some of the above embodiments, the DAR of the antibody-drug conjugate is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0183] In some embodiments, the antibody-drug conjugates provided in this disclosure, their stereoisomers, or pharmaceutically acceptable salts thereof, may also be represented by the structure shown in Formula II: II, Wherein, the anti-ROR1 antibody or its antigen-binding fragment is as described in this disclosure; R 1 Selected from hydrogen atoms, deuterium atoms, and optionally substituted C atoms 1-6 Alkyl groups, optionally substituted C 3-7 cycloalkyl, optionally substituted 3- to 7-membered heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted 5 to 12 heteroaryl groups, R 2 Selected from hydrogen atoms, deuterium atoms, and optionally substituted C atoms 1-6 Alkyl groups, optionally substituted C 3-7 cycloalkyl, optionally substituted 3- to 7-membered heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted 5 to 12 heteroaryl groups; or, R 1 and R 2 Together with the atoms connected to it, they form optional 5- to 8-membered heterocyclic groups that can be substituted.

[0184] In some embodiments, the 3-position of -(succinimide-3-yl-N)- in Formula II is linked to the anti-ROR1 antibody or its antigen-binding fragment via a thioether bond. n has the same meaning as DAR, representing the average number of cytotoxic agents linked to each anti-ROR1 antibody or its antigen-binding fragment. In some embodiments, n is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8. In some embodiments, n is 3.5-5. In some of the above embodiments, n is 3.5-4.5. In some embodiments, n is 7-8. In some embodiments, n is 7.5-8. In some implementations, n is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0185] In some embodiments, the antibody-drug conjugate can be represented by the structure shown in Formula II-1: II-1, Wherein, the anti-ROR1 antibody or its antigen-binding fragment is as described in this disclosure, and n is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8. In some embodiments, n is 3.5-5. In some embodiments, n is 3.5-4.5. In some embodiments, n is 7-8. In some embodiments, n is 7.5-8. In some embodiments, n is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0186] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure are formed by linking a drug-connector of the structure shown in Formula Ia-1 to an anti-ROR1 antibody or its antigen-binding fragment, wherein the drug-connector is attached to the anti-ROR1 antibody or its antigen-binding fragment at the position indicated by * in the structure shown in Formula Ia-1, and the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1 containing the amino acid sequence shown in SEQ ID NO: 25, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 26, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 27, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 28, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 30. In some such embodiments, the DAR of the antibody-drug conjugate is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8; preferably, the DAR of the antibody-drug conjugate is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0187] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure are formed by linking a drug-linker of Formula Ia-1 to an anti-ROR1 antibody or its antigen-binding fragment, wherein the drug-linker is attached to the anti-ROR1 antibody or its antigen-binding fragment at the position indicated by * in the structure of Formula Ia-1, and the anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 60, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 63. The CDRs are defined, for example, according to Kabat, IMGT, Chothia, Contact, AbM, and / or CCG. In some such embodiments, the DAR of the antibody-drug conjugate is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8; preferably, the DAR of the antibody-drug conjugate is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0188] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure are formed by linking a drug-linker of the structure shown in Formula Ia-1 to an anti-ROR1 antibody or its antigen-binding fragment, wherein the drug-linker is attached to the anti-ROR1 antibody or its antigen-binding fragment at the position indicated by * in the structure shown in Formula Ia-1, and the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 60, and a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 63. In some such embodiments, the DAR of the antibody-drug conjugate is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8; preferably, the DAR of the antibody-drug conjugate is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0189] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure are formed by linking a drug-connector of the structure shown in Formula Ia-1 to an anti-ROR1 antibody or its antigen-binding fragment, wherein the drug-connector is attached to the anti-ROR1 antibody or its antigen-binding fragment at the position indicated by * in the structure shown in Formula Ia-1, and the anti-ROR1 antibody or its antigen-binding fragment comprises: a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 60, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 63. In some such embodiments, the DAR of the antibody-drug conjugate is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8; preferably, the DAR of the antibody-drug conjugate is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0190] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure are wherein the antibody-drug conjugate is formed by linking a drug-linker of the structure shown in Formula Ia-1 to an anti-ROR1 antibody, the drug-linker being attached to the anti-ROR1 antibody at the position indicated by * in the structure shown in Formula Ia-1, the anti-ROR1 antibody comprising a heavy chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 83, and a light chain having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with respect to the amino acid sequence shown in SEQ ID NO: 89. In some such embodiments, the DAR of the antibody-drug conjugate is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8; preferably, the DAR of the antibody-drug conjugate is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0191] In some embodiments, the antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof provided in this disclosure is formed by linking a drug-linker of the structure shown in Formula Ia-1 to an anti-ROR1 antibody, wherein the drug-linker is attached to the anti-ROR1 antibody at the position indicated by * in the structure shown in Formula Ia-1, and the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 83, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 89. In some such embodiments, the DAR of the antibody-drug conjugate is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8; preferably, the DAR of the antibody-drug conjugate is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0192] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure are represented by the structure shown in Formula II-1, where n is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8. The anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1 containing the amino acid sequence shown in SEQ ID NO: 25, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 26, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 27, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 28, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 30. In some implementations, n is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0193] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure are represented by the structure shown in Formula II-1, where n is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8. The anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 60, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 63. The CDRs are defined, for example, according to Kabat, IMGT, Chothia, Contact, AbM, and / or CCG. In some implementations, n is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0194] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure are represented by the structure shown in Formula II-1, where n is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8. The anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 60, and an amino acid sequence having ... The amino acid sequence shown in 63 has a light chain variable region with at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity. In some embodiments, n is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0195] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure are represented by the structure shown in Formula II-1, where n is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8. The anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 60, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 63. In some implementations, n is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0196] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure are represented by the structure shown in Formula II-1, where n is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8. The anti-ROR1 antibody comprises a heavy chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 83, and a light chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 89. In some implementations, n is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0197] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure are represented by the structure shown in Formula II-1, where n is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8, and the anti-ROR1 antibody comprises: a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 83, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 89. In some implementations, n is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0198] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure have a DAR of 3.5-4.5 or 4, and the antibody-drug conjugates having a mass fraction of not less than 40%, not less than 45%, not less than 50%, not less than 65%, not less than 70%, not less than 75%, not less than 80%, not less than 85%, not less than 90%, not less than 95%, or not less than 99% for each antibody or its antigen-binding fragment linked to four cytotoxic drugs.

[0199] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure, wherein the DAR of the antibody-drug conjugate is 7-8, 7.5-8, or 8, and the mass fraction of the antibody-drug conjugates to which each antibody or its antigen-binding fragment is linked to 8 cytotoxic drugs is not less than 40%, not less than 45%, not less than 50%, not less than 65%, not less than 70%, not less than 75%, not less than 80%, not less than 85%, not less than 90%, not less than 95%, or not less than 99%.

[0200] In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof provided in this disclosure exhibit one or more combinations of the following properties: (1) Combined with human ROR1, preferably with 2E-09 M or smaller K. D Value combined with ROR1; (2) Combined with monkey ROR1; (3) Internalization occurs in cells expressing ROR1; (4) It has killing activity against tumor cells expressing ROR1; and (5) It has the bystander effect.

[0201] In some embodiments, the antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof has the properties (1) to (5).

[0202] The antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts provided in this disclosure achieve excellent antitumor efficacy and / or safety. In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts exhibit good killing activity against a variety of ROR1-expressing tumor cells. In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts have good in vivo antitumor activity. In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts have excellent safety. In some embodiments, the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts are not prone to aggregation.

[0203] Pharmaceutical Composition In one aspect, this disclosure provides a pharmaceutical composition comprising the antibody-drug conjugate of this disclosure, its stereoisomer, or a pharmaceutically acceptable salt thereof. In some embodiments, the pharmaceutical composition further comprises a pharmaceutically acceptable excipient. Pharmaceutically acceptable excipients include, for example, excipients, diluents, encapsulating materials, fillers, buffers, or other reagents.

[0204] In some embodiments, the pharmaceutical composition is an injectable formulation. In some embodiments, the pharmaceutical composition is an aqueous injectable formulation, including but not limited to aqueous formulations that are not lyophilized or aqueous formulations reconstituted from lyophilized powder. In some embodiments, the pharmaceutical composition is intended for intravenous administration, such as for intravenous injection or intravenous infusion.

[0205] use This disclosure provides for the use of the antibody-drug conjugates of this disclosure, their stereoisomers, or pharmaceutically acceptable salts thereof. This disclosure also provides for the use of the pharmaceutical compositions of this disclosure.

[0206] In one aspect, this disclosure provides the use of the antibody-drug conjugates of this disclosure, their stereoisomers, or pharmaceutically acceptable salts thereof in the preparation of medicaments for treating diseases expressing ROR1.

[0207] In one aspect, this disclosure provides the use of the pharmaceutical compositions of this disclosure in the preparation of a medicament for treating diseases expressing ROR1.

[0208] In one aspect, this disclosure provides the use of the antibody-drug conjugates of this disclosure, their stereoisomers or pharmaceutically acceptable salts thereof, and one or more additional therapeutic agents in the preparation of a medicament for treating diseases expressing ROR1.

[0209] In one aspect, this disclosure provides the use of the pharmaceutical compositions of this disclosure and one or more additional therapeutic agents in the preparation of a medicament for treating diseases expressing ROR1.

[0210] In one aspect, this disclosure provides a method of treating a disease expressing ROR1, the method comprising administering to a subject an antibody-drug conjugate, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof of the present disclosure. In another aspect, this disclosure provides a method of treating a disease expressing ROR1, the method comprising administering to a subject a pharmaceutical composition thereof of the present disclosure. In some embodiments, the antibody-drug conjugate, a stereoisomer thereof, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof of the present disclosure is administered to a subject in a therapeutically effective amount.

[0211] In one aspect, this disclosure provides a method of treating a disease expressing ROR1, the method comprising administering to a subject an antibody-drug conjugate of the present disclosure, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and one or more additional therapeutic agents. In some embodiments, the antibody-drug conjugate of the present disclosure, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, and one or more additional therapeutic agents are administered to the subject in a therapeutically effective amount.

[0212] In one aspect, this disclosure provides a method of treating a disease expressing ROR1, the method comprising administering to a subject a pharmaceutical composition of the present disclosure and one or more additional therapeutic agents. In some embodiments, the pharmaceutical composition of the present disclosure and one or more additional therapeutic agents are administered to the subject in a therapeutically effective amount.

[0213] In some embodiments, the disease expressing ROR1 is a tumor. In some embodiments, the additional therapeutic agent may be a tumor therapeutic agent, such as those known in the art.

[0214] In some embodiments, the method includes contacting tumor cells with the antibody-drug conjugate, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition, thereby killing tumor cells or inhibiting tumor cell growth.

[0215] In some embodiments, the method includes contacting tumor cells with the antibody-drug conjugate, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition, and simultaneously or sequentially contacting the tumor cells with one or more additional therapeutic agents, thereby killing the tumor cells or inhibiting tumor cell growth.

[0216] In some embodiments, administration of the disclosed antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, or the disclosed pharmaceutical composition to a subject can kill tumor cells or inhibit tumor cell growth. In some embodiments, administration of a therapeutically effective amount of the disclosed antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, or the disclosed pharmaceutical composition to a subject can kill tumor cells or inhibit tumor cell growth.

[0217] In some embodiments, administering the antibody-drug conjugate of this disclosure, its stereoisomers, or pharmaceutically acceptable salts thereof, along with one or more other therapeutic agents, to a subject can kill tumor cells or inhibit tumor cell growth. In some embodiments, administering a therapeutically effective amount of the antibody-drug conjugate of this disclosure, its stereoisomers, or pharmaceutically acceptable salts thereof, along with one or more other therapeutic agents, to a subject can kill tumor cells or inhibit tumor cell growth. In some embodiments, administering the pharmaceutical composition of this disclosure, along with one or more other therapeutic agents, to a subject can kill tumor cells or inhibit tumor cell growth. In some embodiments, administering a therapeutically effective amount of the pharmaceutical composition of this disclosure, along with one or more other therapeutic agents, to a subject can kill tumor cells or inhibit tumor cell growth.

[0218] In some embodiments, the tumor is a ROR1-positive tumor. In some embodiments, the tumor is a non-solid tumor. In some embodiments, the tumor is a hematologic malignancy. In some embodiments, the tumor is a solid tumor. In some embodiments, the tumor is leukemia, lymphoma, multiple myeloma, breast cancer, ovarian cancer, ovarian teratoma, colorectal cancer, colorectal cancer, rectal cancer, colon cancer, gastric cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), skin cancer, pancreatic cancer, testicular cancer, bladder cancer, uterine cancer, prostate cancer, or adrenal cancer. In some embodiments, the lymphoma is mantle cell lymphoma (MCL).

[0219] Reagent test kit This disclosure provides a kit comprising the antibody-drug conjugate of this disclosure, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of this disclosure.

[0220] This disclosure describes kits comprising the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions described herein. The kits can be used to implement the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions described herein, or for other uses. In some embodiments, the kit may include the antibody-drug conjugates, their stereoisomers, or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions described herein; optionally, the kit may also include instructions for use. The kit may also include other materials required from a commercial and user perspective, such as other buffers, diluents, needles, syringes, etc.

[0221] Connector - Payload In some respects, this disclosure provides a linker-payload with the structure shown in Formula I: I, Among them, R 1 Selected from hydrogen atoms, deuterium atoms, and optionally substituted C atoms 1-6 Alkyl groups, optionally substituted C 3-7 cycloalkyl, optionally substituted 3- to 7-membered heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted 5 to 12 heteroaryl groups, R 2 Selected from hydrogen atoms, deuterium atoms, and optionally substituted C atoms 1-6 Alkyl groups, optionally substituted C 3-7 cycloalkyl, optionally substituted 3- to 7-membered heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted 5 to 12 heteroaryl groups; or, R 1 and R2 Together with the atoms connected to it, they form optional 5- to 8-membered heterocyclic groups that can be substituted.

[0222] In some implementations, the R 1 and R 2 Each is independently selected from hydrogen atoms or C atoms. 1-5 Alkyl (preferably C) 1-4 Alkyl, such as C 1-3 Alkyl group). In some embodiments, the R... 1 and R 2 Each is independently selected from hydrogen atoms, methyl, ethyl, propyl, or isopropyl. In some embodiments, the R... 1 and R 2 All are hydrogen atoms.

[0223] In one specific embodiment, this disclosure provides a joint-load capacity with the structure shown in Formula I-1: I-1.

[0224] Preparation method This disclosure provides a method for preparing the antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, comprising treating an anti-ROR1 antibody or its antigen-binding fragment under reducing conditions, followed by reacting the anti-ROR1 antibody or its antigen-binding fragment with a linker-loador having the structure shown in Formula I, wherein R 1 Selected from hydrogen atoms, deuterium atoms, and optionally substituted C atoms 1-6 Alkyl groups, optionally substituted C 3-7 cycloalkyl, optionally substituted 3- to 7-membered heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted 5 to 12 heteroaryl groups, R 2 Selected from hydrogen atoms, deuterium atoms, and optionally substituted C atoms 1-6 Alkyl groups, optionally substituted C 3-7 cycloalkyl, optionally substituted 3- to 7-membered heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted 5 to 12 heteroaryl groups; or, R 1 and R 2 Together with the atoms connected to it, they form optional 5- to 8-membered heterocyclic groups that can be substituted.

[0225] In some implementations, the R 1 and R 2 Each is independently selected from hydrogen atoms or C atoms. 1-5 Alkyl (preferably C) 1-4 Alkyl, such as C 1-3 Alkyl group). In some embodiments, the R... 1 and R2 Each is independently selected from hydrogen atoms, methyl, ethyl, propyl, or isopropyl. In some embodiments, the R... 1 and R 2 It is a hydrogen atom.

[0226] In some embodiments, the method includes treating an anti-ROR1 antibody or its antigen-binding fragment under reducing conditions, followed by reacting the anti-ROR1 antibody or its antigen-binding fragment with a linker-loador selected from the structure shown in Formula I-1.

[0227] In some implementations, the reduction condition is in the presence of TCEP.

[0228] Explanation and definition Unless otherwise stated, the following terms as used in this disclosure have the following meanings. A particular term should not be considered uncertain or unclear unless specifically defined, but should be understood in accordance with its ordinary meaning in the art. When a trade name appears herein, it is intended to refer to the corresponding product or its active ingredient.

[0229] The term “optional” or “optionally” means that the event or situation described below may or may not occur, including both the occurrence and non-occurrence of the event or situation.

[0230] The term "substituted" refers to the substitution of one or more hydrogen atoms on a specific atom by a substituent, provided that the valence state of the specific atom is normal and the resulting compound is stable. When the substituent is oxo (i.e., =O), it means that two hydrogen atoms are substituted; oxoation does not occur on the aromatic group. "Optionally substituted" means that it may or may not be substituted, unless otherwise specified; the type and number of substituents can be arbitrary on a chemically feasible basis.

[0231] When any variable (such as R) appears more than once in the composition or structure of a compound, its definition is independent in each case. Therefore, for example, if a group is replaced by two Rs, each R has an independent option.

[0232] The term "thiol" refers to the -SH group.

[0233] Unless otherwise specified, use wedge-shaped solid line keys ( ) and wedge-shaped dashed key ( ) represents the absolute configuration of a solid center.

[0234] Unless otherwise specified, when a group has a connectable site, the connection position of that site with other groups can be indicated by an asterisk (*) or a hash (#). For example, the structure of "-(succinimide-3-yl-N)-" as used herein is shown in the following formula: , where * and # represent the connection of the carbon atom at position 3 and the nitrogen atom at position 1 of the structure to other groups, respectively.

[0235] As used in this article, compounds formed by replacing atoms or groups of atoms in a parent compound molecule with other atoms or groups of atoms are called "derivatives" of the parent compound.

[0236] The compounds disclosed herein may exist in specific geometric or stereoisomer forms. This disclosure envisions all such compounds, including cis and trans isomers, levorotatory and dextrorotatory isomers, (R)- and (S)-enantiomers, diastereomers, (D)- isomers, (L)- isomers, and racemic mixtures thereof, as well as other mixtures, such as mixtures enriched with enantiomers or diastereomers, all of which are within the scope of this disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All such isomers and mixtures thereof are included within the scope of this disclosure.

[0237] Unless otherwise stated, the terms "cis-trans isomers" or "geometric isomers" arise because the single bonds of double bonds or cyclic carbon atoms cannot rotate freely.

[0238] Unless otherwise stated, the term "enantiomer" refers to stereoisomers that are mirror images of each other.

[0239] Unless otherwise stated, the term "diastereomer" refers to a stereoisomer of a molecule having two or more chiral centers and being in a non-mirror relationship with each other.

[0240] The compounds and intermediates disclosed herein may also exist in different tautomer forms, and all such forms are included within the scope of this disclosure. The terms "tautomer" or "tautomer form" refer to structural isomers of different energies that can interconvert via low energy barriers. For example, proton tautomers (also known as proton transfer tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine isomerization. A specific example of a proton tautomer is the imidazole moiety, in which a proton can migrate between two ring nitrogens. Valence tautomers include interconversions via the recombination of some bonding electrons.

[0241] The term "bystander effect," as used herein, refers to an effect in which a cytotoxic drug coupled to an antibody or its antigen-binding fragment via a cleavable or uncleavable linker is able to diffuse across the cell membrane after release from the antibody or its antigen-binding fragment, thereby causing the killing of adjacent cells. The ability to diffuse across the cell membrane is related to the hydrophobicity of the cytotoxic drug or the combination of the cytotoxic drug and the linker. Such cytotoxic drugs can be, for example, eribulin or derivatives thereof. The bystander effect may be desirable, particularly in tumors with heterogeneous target expression and in solid tumors where antibody penetration may be limited.

[0242] The term "treatment" means administering the compounds or pharmaceutical compositions described in this disclosure to improve or eliminate a disease or one or more symptoms associated with said disease, and includes, but is not limited to: (i) Suppress the disease or disease state, that is, curb its development; (ii) To alleviate a disease or disease state, even if the disease or disease state subsides; (iii) To reduce any direct or indirect pathological consequences of disease or disease state.

[0243] The term "therapeutic effective amount" means (i) the amount of the disclosed compound used to treat a particular disease, condition, or disorder, or (ii) to reduce, improve, or eliminate one or more symptoms of a particular disease, condition, or disorder. The amount of the disclosed compound or pharmaceutical composition constituting a "therapeutic effective amount" can vary depending on factors such as the compound or pharmaceutical composition and its ability to elicit a desired response in an individual, the disease state and its severity, the route of administration, and the age, sex, and weight of the mammal to be treated. Therapeutic effective amounts can also routinely be determined by those skilled in the art based on their own knowledge and the content of this disclosure.

[0244] The term "pharmaceutical acceptable" refers to compounds, materials, compositions, and / or dosage forms that, within the bounds of reliable medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0245] The term "pharmaceutically acceptable salt" refers to a salt of a compound (such as the antibody-drug conjugate of this disclosure) that is safe and effective when used in mammals and has the intended biological activity. For example, it may be a metal salt, an ammonium salt, a salt formed with an organic base, a salt formed with an inorganic acid, a salt formed with an organic acid, or a salt formed with a basic or acidic amino acid.

[0246] The term "excipient" refers to any component other than the active ingredient (e.g., the antibody-drug conjugate of this disclosure). The selection of excipients will largely depend on factors such as the specific route of administration, the efficacy of the excipient in terms of solubility and stability, and the nature of the dosage form.

[0247] The term "antibody" is used in the broadest sense, including but not limited to various antibody structures such as monoclonal antibodies, polyclonal antibodies, and multispecific antibodies (e.g., bispecific antibodies, trispecific antibodies), as long as they exhibit the desired antigen-binding activity.

[0248] The term “antigen-binding fragment” refers to one or more fragments of an antibody that retain the function of specifically binding to an antigen. Examples covered by the term “antigen-binding fragment” include: (i) Fab fragments: monovalent fragments consisting of VL, VH, CL, and CH1 domains; (ii) F(ab')2 fragments, bivalent fragments containing two Fab fragments connected by disulfide bonds in the hinge region; (iii) Fd fragments consisting of VH and CH1 domains; (iv) Fv fragments consisting of VL and VH domains of an antibody single arm; (v) dAb fragments consisting of VH domains (see Ward et al., Nature. 341:544-546 (1989)); (vi) separated complementarity-determining regions (CDRs); (vii) nanobodies; (viii) single-chain Fv (scFv); (viiii) scFab.

[0249] The anti-ROR1 antibody or its antigen-binding fragment disclosed herein may be an IgG1, IgG2, IgG3, or IgG4 isotype. The term "isotype" refers to the type of antibody encoded by the heavy chain constant region gene. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment disclosed herein is a human IgG1, human IgG2, human IgG3, or human IgG4 isotype. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment disclosed herein is an IgG1 isotype. In some embodiments, the anti-ROR1 antibody or its antigen-binding fragment disclosed herein is a human IgG1 isotype. The anti-ROR1 antibody or its antigen-binding fragment disclosed herein may be derived from any species, including but not limited to mice, rats, rabbits, non-human primates (such as chimpanzees, cynomolgus monkeys, spider monkeys, rhesus monkeys), llamas, and humans. The anti-ROR1 antibody or its antigen-binding fragment disclosed herein may be a murine antibody, a chimeric antibody, a humanized antibody, or a human antibody.

[0250] The term "mouse antibody" or "mouse-derived antibody" refers to an antibody in which both the backbone region and the CDR region in the variable region are derived from mouse germline immunoglobulin sequences. Furthermore, if the antibody contains a constant region, that constant region is also derived from mouse germline immunoglobulin sequences. The murine antibodies of this disclosure may include amino acid residues not encoded by mouse germline immunoglobulin sequences (e.g., mutations introduced through in vitro random or point mutations or through in vivo somatic mutations), but "mouse antibody" or "mouse-derived antibody" does not include antibodies in which a CDR sequence derived from other mammalian germlines is inserted into the mouse backbone sequence.

[0251] Chimeric antibodies are antibodies formed by fusing the variable region of a murine antibody with the constant region of a human antibody. They can reduce the immune response induced by murine antibodies. To create chimeric antibodies, firstly, a hybridoma that secretes murine-specific monoclonal antibodies is established. Then, the variable region gene is cloned from the hybridoma cells. Next, the constant region gene of the human antibody is cloned as needed. The murine variable region gene and the human constant region gene are then linked to form a chimeric gene, which is inserted into an expression vector. Finally, the chimeric antibody is expressed in a eukaryotic or prokaryotic system.

[0252] "Humanized antibody" is an antibody that contains a complementarity-determining region (CDR) derived from a non-human antibody, a backbone region derived from a human antibody, and a constant region.

[0253] The term "CDR" (complementarity-determining region) is also known as the "hypervariant region." Naturally occurring four-chain antibodies typically contain six CDRs: three in the heavy chain variable region and three in the light chain variable region.

[0254] The term "variable region" refers to the main antigen recognition domain of about 100 to 110 or more amino acids defined by the N-terminal domain of the light or heavy chain of an antibody. The terms light chain variable region (VL) and heavy chain variable region (VH) refer to these light chain domains and heavy chain domains, respectively.

[0255] As used in this article, the term "EC" 50 "Colony" refers to the half-maximal effective concentration, which is the antibody concentration at which the induced response is 50% of the maximum response, i.e., half the distance between the maximum response and the baseline. 50 It can be measured by ELISA or FACS analysis or any other method known in the art.

[0256] “K D "Refers to the equilibrium dissociation constant, which is the self-dissociation rate constant (k d ) relative to the binding rate constant (k a (i.e., k) d / k a The ratio of K to α is expressed as molar concentration (M). The K of the antibody DThe value can be determined using methods well-established in the art. The K value used for antibody determination... D The preferred method is to use surface plasmon resonance (SPR) technology, preferably using a biosensor system such as the Biacore surface plasmon resonance system for analysis.

[0257] The term "identity," also known as consistency, refers to the percentage of amino acid residues in the sequence to be aligned that are identical to those in the specific amino acid sequence shown herein, after aligning the sequence to be aligned with it and, if necessary, introducing vacancies to achieve the maximum percentage of sequence identity, and without considering any conserved substitutions as part of the sequence identity. Amino acid sequence alignment for identity can be performed using various methods within the art, such as BLAST, BLAST-2, ALIGN, or Megalign (DNASTAR) software. Those skilled in the art can determine the appropriate parameters for the aligned sequences, including any algorithm required to achieve maximum alignment across the full length of the compared sequences.

[0258] The term "subject" includes any human or non-human animal. The term "non-human animal" includes all vertebrates, such as mammals and non-mammals, including non-human primates, sheep, dogs, cats, horses, cattle, chickens, amphibians, reptiles, etc. Preferably, the subject according to this disclosure is a human. Unless otherwise stated, the terms "patient" or "subject" may be used interchangeably.

[0259] As used herein, “about” means within the acceptable range of error for a particular value as determined by one of ordinary skill in the art, which depends in part on how the value is measured or determined, i.e., the limitations of the measurement system. For example, “about” may, in accordance with art practice, mean within one or more standard deviations. Alternatively, “about” may mean a range of up to ±5%, such as fluctuations within ±2%, ±1%, or ±0.5% of a given specific numerical range. When a particular value is given in the scope of this disclosure, unless otherwise stated, “about” shall be understood to mean within the acceptable range of error for that particular value. In this document, unless otherwise stated, the values ​​of step parameters or conditions are implicitly modified by “about”.

[0260] The terms “comprise,” “containing,” and “comprising” and their equivalents (e.g., contain, contains, containing, include, includes, and including) shall be understood as “including but not limited to,” meaning that in addition to the listed elements, components, and steps, other unspecified elements, components, and steps may also be covered.

[0261] In this document, unless the context clearly indicates otherwise, singular terms encompass plural referents, and vice versa. Attached Figure Description

[0262] Figure 1A-1C The binding curves of the anti-ROR1 chimeric antibody to ROR1-expressing cells were detected by FACS, in which... Figure 1A For A549 cells, Figure 1B Jeko-1 cells, Figure 1C SW620 cells; Figure 2A-2I The binding curves of anti-ROR1 humanized antibody and ROR1-expressing cells were detected by FACS, in which... Figure 2A , 2D And 2G are PA-1 cells, Figure 2B , 2E 2H represents Jeko-1 cells. Figure 2C , 2F 2I were SW620 cells; Figures 3A-3C To detect the internalization effect of anti-ROR1 humanized antibody in ROR1-expressing cells using FACS, among which... Figure 3A The cells were NCI-H1993-ROR1. Figure 3B MDA-MB-468-ROR1 cells, Figure 3C NCI-H1975-ROR1 cells; Figure 4 The inhibition rate of the non-canonical Wnt signaling pathway stimulated by the anti-ROR1 humanized antibody on Wnt5a in 293T-ROR1-NF-κB cells; Figure 5 The ADCC effect of anti-ROR1 humanized antibody on PA-1 cells; Figures 6A-6F Binding curves of anti-ROR1 antibody-drug conjugates hzAb58-2.1-Eribulin-4(2), hzAb58-2.1-Eribulin-4(1), hzAb58-2.1-Eribulin-8(1) and reference MK-2140 with cells expressing different levels of ROR1 were detected by FACS. Figure 6A For NCI-N87 cells, Figure 6B PA-1 cells, Figure 6C For NCI-H1975 cells, Figure 6D For A549 cells, Figure 6E Jeko-1 cells, Figure 6F SW620 cells; Figures 7A-7FThe internalization effects of anti-ROR1 antibody-drug conjugates hzAb58-2.1-Eribulin-4(2), hzAb58-2.1-Eribulin-8(1), and reference MK-2140 in ROR1-expressing cells were detected by FACS. Figure 7A MDA-MB-468-ROR1 cells, Figure 7B For NCI-N87 cells, Figure 7C PA-1 cells, Figure 7D For NCI-H1975 cells, Figure 7E For A549 cells, Figure 7F Jeko-1 cells; Figures 8A-8E The killing rate curves of hzAb58-2.1-Eribulin-4(1), hzAb58-2.1-Eribulin-4(2), hzAb58-2.1-Eribulin-8(1) and the reference product MK-2140 against ROR1-positive tumor cells are shown. Figure 8A The cells were NCI-H1975-ROR1. Figure 8B MDA-MB-468-ROR1 cells, Figure 8C For NCI-N87 cells, Figure 8D Jeko-1 cells, Figure 8E PA-1 cells; Figure 9 The bystander effect of hzAb58-2.1-Eribulin-4(2), hzAb58-2.1-Eribulin-8(1) and reference MK-2140 was detected by FACS method, with NCI-H1975-ROR1 as positive cells, Jurkat as negative cells, and no ADC added in the negative control group; Figure 10 The relative tumor volume curves of the xenografts after administration of hzAb58-2.1-Eribulin-4(1), hzAb58-2.1-Eribulin-8(1), reference product MK-2140, and solvent control were obtained in a human non-small cell lung cancer NCI-H1975 nude mouse subcutaneous xenograft model. Figure 11A-11B The killing rate curves of hzAb19-3.2-Eribulin-DAR4(1) and hzAb56-1-1.1-Eribulin-DAR4(1) against ROR1-positive tumor cells are shown. Figure 11A PA-1 cells, Figure 11B Jeko-1 cells. Detailed Implementation

[0263] This disclosure also provides the following specific implementation schemes, but the scope of protection of this disclosure is not limited thereto: Implementation Scheme 1. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, wherein the antibody-drug conjugate comprises an anti-ROR1 antibody or an antigen-binding fragment thereof, wherein the anti-ROR1 antibody or the antigen-binding fragment thereof comprises: HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 1, 9, 17, or 25; HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 2, 10, 18, 26, 49, or 50; HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 3, 11, 19, or 27; LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 4, 12, 20, or 28; LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 5, 13, 21, or 29; and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 6, 14, 22, or 30.

[0264] Implementation Scheme 2. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to Implementation Scheme 1, wherein the anti-ROR1 antibody or its antigen-binding fragment comprises: (1) HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 1, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 2, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 3, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 4, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 5, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 6; (2) HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 50, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14; SEQ ID NO: In 50, X1 is selected from S or A, X2 is selected from A or Q, and X3 is selected from K or Q; (3) HCDR1 containing the amino acid sequence shown in SEQ ID NO: 9, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 10, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 11, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 12, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 14; (4) HCDR1 containing the amino acid sequence shown in SEQ ID NO: 9, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 49, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 11, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 12, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 14; (5) HCDR1 containing the amino acid sequence shown in SEQ ID NO: 17, containing the amino acid sequence shown in SEQ ID NO: 18, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 19, and LCDR2 containing the amino acid sequence shown in SEQ ID NO: 19, and LCDR3 ...2 containing the amino acid sequence shown in SEQ ID NO: 19 HCDR2 containing the amino acid sequence shown in SEQ ID NO:19, HCDR3 containing the amino acid sequence shown in SEQ ID NO:20, LCDR1 containing the amino acid sequence shown in SEQ ID NO:21, and LCDR3 containing the amino acid sequence shown in SEQ ID NO:22.Or (6) HCDR1 containing the amino acid sequence shown in SEQ ID NO: 25, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 26, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 27, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 28, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 30.

[0265] Implementation Scheme 3. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, wherein the antibody-drug conjugate comprises an anti-ROR1 antibody or an antigen-binding fragment thereof, wherein the anti-ROR1 antibody or the antigen-binding fragment thereof comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 7, 15, 23, 31, 51, 52, 56, 57, 58, 60, or 61, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 8, 16, 24, 32, 53, 54, 55, 59, 62, 63, or 64.

[0266] Implementation Scheme 4. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to Implementation Scheme 3, wherein the anti-ROR1 antibody or its antigen-binding fragment comprises: (1) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 7, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 8; (2) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 15, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 16; (3) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 23, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 24; (4) HCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 25. (5) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region shown in SEQ ID NO: 51, and LCDR1, LCDR2, and LCDR3 in the light chain variable region shown in SEQ ID NO: 53; (6) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region shown in SEQ ID NO: 51, and LCDR1, LCDR2, and LCDR3 in the light chain variable region shown in SEQ ID NO: 54; (7) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region shown in SEQ ID NO: 51, and LCDR1, LCDR2, and LCDR3 in the light chain variable region shown in SEQ ID NO: 55; (8) HCDR1, HCDR2, and HCDR3 in the light chain variable region shown in SEQ ID NO: 55. (9) HCDR1, HCDR2 and HCDR3 in the heavy chain variable region shown in SEQ ID NO: 52, and LCDR1, LCDR2 and LCDR3 in the light chain variable region shown in SEQ ID NO: 53;(10) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 52, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 55; (11) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 56, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 59; (12) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 57, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 59; (13) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 58, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 59. (14) The amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 60, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 62; (15) The amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 60, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 63; (16) The amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 60, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 64; (17) The amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 61, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 64. (18) The amino acid sequences of HCDR1, HCDR2, and HCDR3 in the light chain variable region shown in SEQ ID NO: 61, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 in the light chain variable region shown in SEQ ID NO: 63; or (19) The amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region shown in SEQ ID NO: 61, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 in the light chain variable region shown in SEQ ID NO: 64.

[0267] Implementation Scheme 5. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of Implementation Schemes 1-4, wherein the anti-ROR1 antibody or its antigen-binding fragment is a monoclonal antibody, a monospecific antibody, a multispecific antibody, a Fab fragment, an F(ab')2 fragment, an Fd fragment, an Fv fragment, a dAb, an isolated CDR, scFv, scFab, or a nanobody.

[0268] Implementation Scheme 6. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of Implementation Schemes 1-5, wherein the anti-ROR1 antibody or antigen-binding fragment is murine, chimeric, or humanized.

[0269] Implementation Scheme 7. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of Implementation Schemes 1-6, wherein the anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 7, 15, 23, 31, 51, 52, 56, 57, 58, 60, or 61.

[0270] Implementation Scheme 8. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of Implementation Schemes 1-7, wherein the anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 8, 16, 24, 32, 53, 54, 55, 59, 62, 63, or 64.

[0271] Implementation Scheme 9. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of Implementation Schemes 1-6, wherein the anti-ROR1 antibody or its antigen-binding fragment comprises: (1) a heavy chain variable region having an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 7, and a light chain variable region having an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 8; (2) a heavy chain variable region having an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 8; The amino acid sequence shown in SEQ ID NO: 15 has a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 16; (3) The amino acid sequence has a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 23; and the amino acid sequence has a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 23. The amino acid sequence shown in SEQ ID NO: 31 has a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical; (4) a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 32;(5) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 53; (6) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 53; The amino acid sequence shown in SEQ ID NO: 54 has a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 51; (7) a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 55; and a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 55; (8) an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 55; The amino acid sequence shown in SEQ ID NO: 52 has a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 53;(9) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 52, and a light chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 54; (10) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 52, and a light ... The amino acid sequence shown in SEQ ID NO: 55 has a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 56; (11) a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 59; and a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 59; (12) an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 59. The amino acid sequence shown in SEQ ID NO: 57 has a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 59;(13) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 58, and a light chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 59; (14) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 60, and a light ... The amino acid sequence shown in SEQ ID NO: 62 has a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 60; (15) a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 63; and a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 63; (16) an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 63; The amino acid sequence shown in SEQ ID NO: 60 has a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 64;(17) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 62; (18) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 62; The amino acid sequence shown in SEQ ID NO: 63 has a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 61; or (19) a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 64;

[0272] Implementation Scheme 10. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of Implementation Schemes 1-6, wherein the anti-ROR1 antibody or its antigen-binding fragment comprises: (1) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 7, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 8; (2) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 15, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 16; (3) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 23, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 24; (4) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 31, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 32; (5) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 53; (6) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 53; (7) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region containing the amino acid sequence shown in SEQ ID NO: 55; (8) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 52, and a light chain variable region containing the amino acid sequence shown in SEQ ID NO: 53; (9) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 52, and a light chain variable region containing the amino acid sequence shown in SEQ ID NO: 54; (10) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 52, and a light chain variable region containing the amino acid sequence shown in SEQ ID NO: 55; (11) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 56, and a light chain variable region containing the amino acid sequence shown in SEQ ID NO: 59; (12) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 57, and a light chain variable region containing the amino acid sequence shown in SEQ ID NO: 54. (13) A light chain variable region containing the amino acid sequence shown in SEQ ID NO: 59; (14) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 60; (15) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 60; and a light chain variable region containing the amino acid sequence shown in SEQ ID NO: 63.(16) A heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 60, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 64; (17) A heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 62; (18) A heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 63; or (19) A heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 64.

[0273] Implementation Scheme 11. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of Implementation Schemes 1-10, wherein the anti-ROR1 antibody is an IgG1, IgG2, IgG3, or IgG4 isotype; preferably, the anti-ROR1 antibody is a human IgG1, human IgG2, human IgG3, or human IgG4 isotype; more preferably, the anti-ROR1 antibody is a human IgG1 isotype.

[0274] Implementation Scheme 12. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of Implementation Schemes 1-11, wherein the anti-ROR1 antibody comprises a heavy chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 33, 37, 41, 45, 65, 67, 75, 77, 79, 83, or 85.

[0275] Implementation Scheme 13. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of Implementation Schemes 1-12, wherein the anti-ROR1 antibody comprises a light chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 35, 39, 43, 47, 69, 71, 73, 81, 87, 89, or 91.

[0276] Implementation Scheme 14. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of Implementation Schemes 1-11, wherein the anti-ROR1 antibody comprises: (1) a heavy chain having an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 33, and a light chain having an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 35; (2) an amino acid sequence having an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 35; (3) A heavy chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity with the amino acid sequence shown in SEQ ID NO: 39, and a light chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity with the amino acid sequence shown in SEQ ID NO: 39; (3) A heavy chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity with the amino acid sequence shown in SEQ ID NO: 41, and a light .... (3) A light chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity with the amino acid sequence shown in SEQ ID NO: 45; (4) A heavy chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity with the amino acid sequence shown in SEQ ID NO: 47;(5) A heavy chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 65, and a light chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 69; (6) A heavy chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 65, and a light ... (7) A light chain whose amino acid sequence is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 65; and a heavy chain whose amino acid sequence is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 73; and a light chain whose amino acid sequence is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 73; The heavy chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 69, and the light chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 69;(9) A heavy chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 67, and a light chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 71; (10) A heavy chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 67, and a light ... The amino acid sequence shown in SEQ ID NO: 73 has at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity with the amino acid sequence shown in SEQ ID NO: 75; (11) a heavy chain with an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity with the amino acid sequence shown in SEQ ID NO: 81; and a light chain with an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity with the amino acid sequence shown in SEQ ID NO: 81; (12) an amino acid sequence having an amino acid sequence having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity with the amino acid sequence shown in SEQ ID NO: 75. The heavy chain shown in SEQ ID NO: 87 has an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 81; and the light chain has an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 81.(13) A heavy chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 79, and a light chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 81; (14) A heavy chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 83, and a light ... (15) A light chain whose amino acid sequence is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence shown in SEQ ID NO: 83; and a heavy chain whose amino acid sequence is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence shown in SEQ ID NO: 89; and a light chain whose amino acid sequence is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence shown in SEQ ID NO: 89; The heavy chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 91, and the light chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 91;(17) A heavy chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 85, and a light chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 87; (18) A heavy chain having an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 85, and a light ... The light chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 85; or (19) the heavy chain having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 91;

[0277] Implementation Scheme 15. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, wherein the antibody-drug conjugate comprises an anti-ROR1 antibody, the anti-ROR1 antibody comprising: (1) a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 33, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 35; (2) a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 37, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 39; (3) a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 41, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 43; (4) a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 45, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 47; (5) a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 65, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 69; (6) a heavy chain comprising the amino acid sequence shown in SEQ ID NO: 65, and a light chain comprising the amino acid sequence shown in SEQ ID NO: 69. (7) A light chain containing the amino acid sequence shown in SEQ ID NO: 65 and a light chain containing the amino acid sequence shown in SEQ ID NO: 73; (8) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 67 and a light chain containing the amino acid sequence shown in SEQ ID NO: 69; (9) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 67 and a light chain containing the amino acid sequence shown in SEQ ID NO: 71; (10) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 67 and a light chain containing the amino acid sequence shown in SEQ ID NO: 73; (11) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 75 and a light chain containing the amino acid sequence shown in SEQ ID NO: 81; (12) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 77 and a light chain containing the amino acid sequence shown in SEQ ID NO: 81; (13) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 79 and a light chain containing the amino acid sequence shown in SEQ ID NO: 71. (14) A light chain containing the amino acid sequence shown in SEQ ID NO: 83 and a light chain containing the amino acid sequence shown in SEQ ID NO: 87; (15) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 83 and a light chain containing the amino acid sequence shown in SEQ ID NO: 89; (16) A heavy chain containing the amino acid sequence shown in SEQ ID NO: 83 and a light chain containing the amino acid sequence shown in SEQ ID NO: 91.(17) A heavy chain comprising the amino acid sequence shown in SEQ ID NO: 85 and a light chain comprising the amino acid sequence shown in SEQ ID NO: 87; (18) A heavy chain comprising the amino acid sequence shown in SEQ ID NO: 85 and a light chain comprising the amino acid sequence shown in SEQ ID NO: 89; or (19) A heavy chain comprising the amino acid sequence shown in SEQ ID NO: 85 and a light chain comprising the amino acid sequence shown in SEQ ID NO: 91.

[0278] Implementation Scheme 16. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of Implementation Schemes 1-15, wherein the antibody-drug conjugate is formed by linking a drug-connector of the structure shown in Formula Ia to an anti-ROR1 antibody or its antigen-binding fragment, wherein the drug-connector is attached to the anti-ROR1 antibody or its antigen-binding fragment at the position indicated by * in the structure shown in Formula Ia. Ia; where R 1 Selected from hydrogen atoms, deuterium atoms, and optionally substituted C atoms 1-6 Alkyl groups, optionally substituted C 3-7 cycloalkyl, optionally substituted 3- to 7-membered heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted 5 to 12 heteroaryl groups, R 2 Selected from hydrogen atoms, deuterium atoms, and optionally substituted C atoms 1-6 Alkyl groups, optionally substituted C 3-7 cycloalkyl, optionally substituted 3- to 7-membered heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted 5 to 12 heteroaryl groups; or, R 1 and R 2 Together with the atoms connected to it, they form optional 5- to 8-membered heterocyclic groups that can be substituted.

[0279] Implementation Scheme 17. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to Implementation Scheme 16, wherein the R 1 and R 2 Each is independently selected from hydrogen atoms or C atoms. 1-5 Alkyl, wherein C 1-5 Alkyl group preferred C 1-4 Alkyl, more preferably C 1-3 alkyl.

[0280] Implementation Scheme 18. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, according to Implementation Scheme 16 or 17, wherein the R 1 and R 2 Each is independently selected from hydrogen atom, methyl, ethyl, propyl or isopropyl.

[0281] Implementation Scheme 19. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, according to any one of Implementation Schemes 16-18, wherein the R 1 and R 2 All are hydrogen atoms.

[0282] Implementation Scheme 20. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, according to any one of Implementation Schemes 1-19, wherein the DAR of the antibody-drug conjugate is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8.

[0283] Implementation Scheme 21. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, according to any one of Implementation Schemes 1-19, wherein the DAR of the antibody-drug conjugate is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0284] Implementation Scheme 22. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of Implementation Schemes 1-19, wherein the structure of the antibody-drug conjugate is shown in Formula II-1 below: II-1, where n is 1-10, 2-8, 3.5-8, 4-8, 3.5-4.5, 3.5-5, 4-5, 6-8, 7-8, or 7.5-8.

[0285] Implementation Scheme 23. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof as described in Implementation Scheme 22, wherein n is 2, 3, 3.5, 3.6, 3.7, 3.8, 3.9, 4, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5, 6, 7, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.

[0286] Implementation Scheme 24. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of Implementation Schemes 1-23, wherein the antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof exhibits one or more of the following properties in combination: binding to human ROR1, preferably binding to human ROR1 with a KD value of 2E-09 M or less; binding to monkey ROR1; internalization in cells expressing ROR1; cytotoxic activity against tumor cells expressing ROR1; and having a bystander effect.

[0287] Implementation Scheme 25. A pharmaceutical composition comprising the antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, as described in any one of Implementation Schemes 1-24; optionally, the pharmaceutical composition further comprising a pharmaceutically acceptable excipient.

[0288] Implementation Scheme 26. A method for preparing the antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof as described in any one of Implementation Schemes 16-24, comprising treating an anti-ROR1 antibody or its antigen-binding fragment under reducing conditions, followed by reacting the anti-ROR1 antibody or its antigen-binding fragment with a linker-loador having the structure shown in Formula I, wherein the structure shown in Formula I is as follows: I,R 1 Selected from hydrogen atoms, deuterium atoms, and optionally substituted C atoms 1-6 Alkyl groups, optionally substituted C 3-7 cycloalkyl, optionally substituted 3- to 7-membered heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted 5 to 12 heteroaryl groups, R 2 Selected from hydrogen atoms, deuterium atoms, and optionally substituted C atoms 1-6 Alkyl groups, optionally substituted C 3-7 cycloalkyl, optionally substituted 3- to 7-membered heterocyclic groups, optionally substituted C 6-10 aryl, optionally substituted 5 to 12 heteroaryl groups; or, R 1 and R 2 Together with the atoms connected to it, they form optional 5- to 8-membered heterocyclic groups that can be substituted.

[0289] Implementation Scheme 27. The method according to Implementation Scheme 26, wherein the R 1 and R 2 Each is independently selected from hydrogen atoms or C atoms. 1-5 Alkyl, wherein C 1-5 Alkyl group preferred C 1-4 Alkyl, more preferably C 1-3 alkyl.

[0290] Implementation Scheme 28. The method according to Implementation Scheme 26 or 27, wherein the R 1and R 2 Each is independently selected from hydrogen atom, methyl, ethyl, propyl or isopropyl.

[0291] Implementation Scheme 29. The method according to any one of Implementation Schemes 26-28, wherein the R 1 and R 2 All are hydrogen atoms.

[0292] Implementation Scheme 30. Use of the antibody-drug conjugate, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of any one of Implementation Schemes 1-24 in the preparation of a medicament for treating diseases expressing ROR1.

[0293] Implementation Scheme 31. Use of the antibody-drug conjugate, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition of Implementation Scheme 25, and one or more additional therapeutic agents in the preparation of a medicament for treating diseases expressing ROR1.

[0294] Implementation Scheme 32. The use according to Implementation Scheme 30 or 31, wherein the disease expressing ROR1 is a tumor.

[0295] Implementation Scheme 33. The use according to Implementation Scheme 31 or 32, wherein the additional therapeutic agent is a tumor therapeutic agent.

[0296] Implementation Scheme 34. The use according to Implementation Scheme 32 or 33, wherein the tumor is a ROR1 positive tumor.

[0297] Implementation Scheme 35. The use according to any one of Implementation Schemes 32-34, wherein the tumor is leukemia, lymphoma (e.g., mantle cell lymphoma (MCL)), multiple myeloma, breast cancer, ovarian cancer, ovarian teratoma, intestinal cancer, colorectal cancer, rectal cancer, colon cancer, gastric cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), skin cancer, pancreatic cancer, testicular cancer, bladder cancer, uterine cancer, prostate cancer, or adrenal cancer.

[0298] Implementation Scheme 36. A method for treating a disease expressing ROR1, comprising administering to a subject the antibody-drug conjugate, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in Implementation Scheme 25.

[0299] Implementation Scheme 37. The method according to Implementation Scheme 36, wherein the disease expressing ROR1 is a tumor.

[0300] Implementation Scheme 38. The method according to Implementation Scheme 37, the method comprising contacting tumor cells with any one of Implementation Schemes 1-24, an antibody-drug conjugate thereof, a stereoisomer thereof or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to Implementation Scheme 25, thereby killing tumor cells or inhibiting tumor cell growth.

[0301] Implementation Scheme 39. The method according to any one of Implementation Schemes 36-38, wherein the method further comprises administering one or more additional therapeutic agents to the subject.

[0302] Implementation Scheme 40. The method according to Implementation Scheme 39, wherein the additional therapeutic agent is a tumor therapeutic agent.

[0303] Implementation Scheme 41. The method according to any one of Implementation Schemes 37-40, wherein the tumor is a ROR1 positive tumor.

[0304] Implementation Scheme 42. The method according to any one of Implementation Schemes 37-41, wherein the tumor is leukemia, lymphoma (e.g., mantle cell lymphoma (MCL)), multiple myeloma, breast cancer, ovarian cancer, ovarian teratoma, intestinal cancer, colorectal cancer, rectal cancer, colon cancer, gastric cancer, lung cancer (including small cell lung cancer and non-small cell lung cancer), skin cancer, pancreatic cancer, testicular cancer, bladder cancer, uterine cancer, prostate cancer, or adrenal cancer.

[0305] Implementation Scheme 43. A kit comprising the antibody-drug conjugate, its stereoisomer or a pharmaceutically acceptable salt thereof, or the pharmaceutical composition described in Implementation Scheme 25; optionally, the kit further comprising instructions for use.

[0306] Example For clarity, this disclosure is further illustrated by embodiments, but these embodiments are not intended to limit the scope of this disclosure. Those skilled in the art will readily identify various non-critical parameters that can be changed or modified to produce substantially similar results.

[0307] The UC-961 used in these examples is Zilovertamab from Oncternal Therapeutics. The isotype IgG used in Examples 8 and 10 is a human IgG1 antibody (Sino Biological, catalog number: HG1K). Cell information related to the examples in this disclosure is shown in the table below:

[0308] Example 1: Preparation of ROR1 antigen Nucleic acid molecules encoding the human ROR1 extracellular domain-human Fc fusion protein (human ROR1 ECD-hFc) or the human / cynomolgus monkey ROR1 ECD-His tagged recombinant protein (human / cynomolgus monkey ROR1 ECD-His) are inserted into the expression vector pcDNA3.1(+) (Invitrogen) at appropriate restriction enzyme sites. ® In Expi293 cells (THERMO FISHER SCIENTIFIC) ® The human ROR1 ECD-hFc and human / cyno ROR1 ECD-His were expressed in (catalog number: 100044202). The amino acid sequence of human ROR1 ECD-hFc is shown in SEQ ID NO: 93; the amino acid sequence of human / cyno ROR1 ECD-His is shown in SEQ ID NO: 94.

[0309] Example 2: Generation of anti-ROR1 monoclonal antibody Human ROR1 ECD-hFc was emulsified with Freund's complete adjuvant or Freund's incomplete adjuvant at a volume ratio of 1:1. 25 μg / mouse was subcutaneously injected into A / J mice, BALB / c mice, or SJL mice every 2-3 weeks for 7-9 weeks. Booster immunizations were initiated from the second immunization onwards. Serum was collected one week after each booster immunization, and antibody titers were determined by ELISA. Mice with serum antibody titers higher than 1:10 after immunization were selected. 5 Cell fusion was performed on mice. Three days prior to cell fusion, mice were immunized via intraperitoneal injection of adjuvant-free human ROR1 ECD-hFc. Electrofusion was performed using a BTX electrofusion apparatus. ® Model: ECM ® (-2001) Fused spleen cells from immunized mice with SP2 / 0 myeloma cells. After fusion, the cells were treated with HAT (Gibco)-containing... ® Catalog number: 21060-017) and serum (Gibco) ® SFM medium (Gibco) for (catalog number: 10099-141C) ® (Catalogue No.: 11995-040) The cells were resuspended and added to a 96-well plate at 100 μL / well. After incubation at 37°C and 5% CO2 for 7 days, the hybridoma cell culture supernatant was collected for screening anti-ROR1 monoclonal antibodies.

[0310] Example 3: Screening of anti-ROR1 monoclonal antibodies Human / cyno ROR1 ECD-His hybridoma cell culture supernatant was used for screening using ELISA to obtain positive clones. Subcloning of the positive clones yielded positive monoclonal clones. The Jeko-1 cell line, which naturally expresses ROR1, was used to evaluate the obtained positive monoclonal clones using flow cytometry (FACS), and the following positive monoclonal clones were selected: m3G9B1, m29H5D8, m135A2B10, and m137G11D10.

[0311] Example 4: Preparation of anti-ROR1 chimeric antibody (1) Acquisition of cDNA Hybridoma cells m3G9B1, m29H5D8, m135A2B10, and m137G11D10 were selected and total RNA was extracted using a Takara total RNA extraction kit. ® Total RNA was isolated from cells as a template (catalog number: 9767) and processed according to the kit instructions (THERMOFISHER SCIENTIFIC). ® (Catalogue No.: 18080051) First-strand cDNA was synthesized using superscript III reverse transcriptase. Using the first-strand cDNA as a template, the variable region sequence of a mouse antibody was amplified by PCR using mouse IgG primers and Kappa primers. The PCR mixture was separated by electrophoresis on a 1% agarose / Tris-borate gel containing 0.5 μg / mL ethidium bromide. DNA fragments of the desired size were excised from the gel and purified. The purified PCR product was cloned into the pMD-19T vector (Takara). ® (Catalogue No.: 6013) and transformed into DH5α competent Escherichia coli cells (Takara). ® The culture was carried out in LB solid culture plates (catalog number: 9057). Single colonies were picked from the plates and DNA sequencing was performed to obtain the heavy chain variable region and light chain variable region sequences of the anti-ROR1 mouse antibody, as shown in Table 1.

[0312] (2) Construction and expression of chimeric antibodies A chimeric light chain expression plasmid was constructed by ligating a mouse VL region gene fragment to a human κ chain constant region gene fragment, and a chimeric heavy chain expression plasmid was constructed by ligating a mouse VH region gene fragment to a human IgG1 constant region gene fragment. When the ExpiCHO-S cell density reached 6 × 10⁻⁶... 6 At a concentration of 100 cells / mL, the chimeric heavy chain expression plasmid and chimeric light chain expression plasmid corresponding to each antibody were co-transfected into ExpiCHO-S cells (THERMO FISHER SCIENTIFIC) at a 1:1 ratio. ®Protein expression was performed using ExpiCHO Expression Medium (THERMO FISHERSCIENTIFIC). Transfected cells were then used to express proteins in the medium. ® (Catalogue No.: A29100-01) was cultured at 37℃ with 8% CO2. After 7-10 days of culture, it was transduced using a Protein A column (GE Healthcare). ® (Catalog No.: 17-5474-02) Purify chimeric antibodies from cell culture supernatant.

[0313] Table 1. Sequence information of anti-ROR1 mouse antibodies and chimeric antibodies (SEQ ID NO.)

[0314] Note: The VH and VL amino acid sequences of the mouse antibody and the chimeric antibody are the same, while the amino acid and nucleotide sequences of the heavy and light chains are the same as those of the heavy and light chains of the anti-ROR1 chimeric antibody.

[0315] Example 5: Detection of in vitro binding activity of anti-ROR1 chimeric antibody (1) Surface plasmon resonance (SPR) technique for detecting the binding of anti-ROR1 chimeric antibody to ROR1 A certain amount of anti-ROR1 antibody was captured using an Anti-hIgG chip, and then human / cynomolgus monkey ROR1 protein (Acro Biosystems) was passed through the chip surface. ® (Catalog number: RO1-H522y), using Biacore ® The reaction signal was monitored in real time to obtain binding and dissociation curves. The buffer solution used in the experiment was Biacore. ® Universal buffer (137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4·12H2O, 1.8 mM KH2PO4, 0.05% surfactant P-20, pH 7.4). Anti-hIgG was conjugated to the CM5 chip surface to capture anti-ROR1 chimeric antibody and benchmark UC-961 (MCE). ® (Catalogue No.: HY-P99201), and then the signal values ​​of the interaction between different concentrations of ROR1 protein (100 nM, 50 nM, 25 nM, 12.5 nM, 6.25 nM, 3.125 nM) and anti-ROR1 antibody were detected. The flow rate of the flow cell was 50 µL / min, the binding time was 240 s, the dissociation time was 1400 s, and regeneration with 3 M MgCl2 (GE) was performed for 60 s, after which the baseline stabilized. According to Biacore ®The affinity results are obtained by combining the affinity and kinetics 1:1 model in evaluationsoftware. The affinity results are shown in Table 2.

[0316] Table 2. Binding affinity of anti-ROR1 chimeric antibody to ROR1

[0317] (2) FACS detection of the binding of anti-ROR1 chimeric antibody to cells Cell lines A549, SW620, and Jeko-1, which naturally express ROR1, were used at a concentration of 1 × 10⁻⁶ cells / year. 5 Cells / well were added to 96-well plates, and serially diluted anti-ROR1 antibody (starting at 6.67 nM, 5-fold dilution, 7 concentrations) was added. The plates were incubated at 4°C for 0.5 h. The 96-well plates were then rinsed with FACS buffer (Miltenyi). ® After washing three times, the second antibody PE anti-human IgG Fc (Jackson) was added. (Catalog No.: 130-091-221) ® (Catalogue No.: 109-116-170) was diluted 1:200 and added to 100 μL / well of a 96-well plate, and incubated at 4°C for 20 min. Cells were washed three times with FACS buffer and analyzed by flow cytometry (Sartorius). ® (Model: iQue3) Detection and analysis. Analyze the data and calculate EC. 50 The results are shown in Table 3 and Figures 1A-1C As shown.

[0318] Table 3. Binding of anti-ROR1 chimeric antibody to ROR1-expressing cells (ECG) 50 and maximum combination value

[0319] Example 6: Humanization of anti-ROR1 antibody Three antibodies with good binding activity, m29H5D8, m135A2B10, and m137G11D10, were selected for humanization. The m29H5D8, m135A2B10, and m137G11D10 antibodies were humanized using the CDR transplantation method. Human germline antibody sequences with high homology to mouse m29H5D8, m135A2B10, and m137G11D10 antibodies were screened. The complementarity-determining regions (CDRs) in the VH and VL regions of the mouse m29H5D8, m135A2B10, and m137G11D10 antibodies were grafted onto the template human antibody. Several amino acid residue sites in the frame region (FR) of the template human antibody VH and VL were reverted to the corresponding amino acid residues in the mouse m29H5D8, m135A2B10, and m137G11D10 antibodies to obtain humanized light chain variable regions and humanized heavy chain variable regions.

[0320] The humanized light chain variable region was ligated to the human κ chain constant region to construct the humanized light chain, and the humanized heavy chain variable region was ligated to the human IgG1 constant region to construct the humanized heavy chain. Humanized antibodies were constructed and expressed according to the method in Example 4. The sequence information of the anti-ROR1 humanized antibodies is shown in Table 4. The humanized antibodies for m29H5D8 are hz29H5D8-1.1, hz29H5D8-1.2, hz29H5D8-2.1, hz29H5D8-2.2, hz29H5D8-3.1, and hz29H5D8-3.2, and the humanized antibodies for m135A2B10 are hz135A2B10-1.1 and hz135A2B10. -1.2 and hz135A2B10-1.3, the humanized antibodies for m137G11D10 are hz137G11D10-1.1, hz137G11D10-1.2, hz137G11D10-2.1, hz137G11D10-2.2, hz137G11D10-3.1 and hz137G11D10-3.2.

[0321] Table 4. Sequence information of anti-ROR1 humanized antibody (SEQ ID NO.)

[0322] Example 7: Detection of binding activity of anti-ROR1 humanized antibody The binding of the anti-ROR1 humanized antibody to ROR1 was detected using SPR, and the binding of the anti-ROR1 humanized antibody to cells naturally expressing ROR1 was detected using FACS. The experimental methods were as described in Example 5. The cells used for detecting the binding of the anti-ROR1 humanized antibody to cells naturally expressing ROR1 were PA-1, Jeko-1, and SW620. The binding affinity results of the anti-ROR1 humanized antibody to ROR1 are shown in Table 5, and the binding results of the anti-ROR1 humanized antibody to cells expressing ROR1 are shown in Tables 6-1 to 6-3. Figures 2A-2I As shown.

[0323] Table 5. Binding affinity of anti-ROR1 humanized antibody to ROR1

[0324] Table 6-1. Binding of anti-ROR1 humanized antibody to cells expressing ROR1 (EC) 50 and maximum combination value

[0325] Table 6-2. Binding of anti-ROR1 humanized antibody to cells expressing ROR1 (EC) 50 and maximum combination value

[0326] Table 6-3. Binding of anti-ROR1 humanized antibody to cells expressing ROR1 (EC) 50 and maximum combination value

[0327] Example 8: Internalization of anti-ROR1 humanized antibody The target gene encoding human ROR1 (SEQ ID NO: 95) was transfected into NCI-1975, NCI-H1993 and MDA-MB-468 cells, respectively, to obtain stable cell lines NCI-1975-ROR1, NCI-H1993-ROR1 and MDA-MB-468-ROR1.

[0328] Anti-ROR1 antibody internalization analysis was performed based on the binding of ROR1 to stable cell lines NCI-1975-ROR1, NCI-H1993-ROR1, and MDA-MB-468-ROR1, with isotype IgG used as a negative control. The cell density of ROR1-expressing cells was adjusted to 2 × 10⁻⁶ cells / year. 6 The assay kit (Sartorius) was prepared by adding 20 μL of the sample per well to a 96-well plate. ®The internalization assay reagent (catalog number: 90564) was mixed with the anti-ROR1 humanized antibody at a 1:1 volume ratio. For NCI-H1975-ROR1 cells, the final concentration of the anti-ROR1 humanized antibody was 26.7 nM; for NCI-H1993-ROR1 and MDA-MB-468-ROR1 cells, the final concentration was 5.3 nM. After incubation at 37°C in the dark for 15 min, six concentration gradients were created by serial dilution (5-fold). Each gradient was then added at 20 μL / well to a 96-well plate and incubated at 37°C for 2-3 h. The data were analyzed using a flow cytometer (Sartorius, iQue3). EC50 was calculated. 50 The result is as follows Figures 3A-3C As shown in Tables 7-1 and 7-2, these data indicate that hz29H5D8-3.2, hz135A2B10-1.1, and hz137G11D10-2.1 can effectively exert internalization effects in cells expressing ROR1.

[0329] Table 7-1. Internalization of anti-ROR1 humanized antibodies in ROR1-expressing cells.

[0330] Table 7-2. Internalization of anti-ROR1 humanized antibodies in ROR1-expressing cells.

[0331] Example 9: Inhibitory effect of anti-ROR1 humanized antibody on Wnt5a-stimulated non-classical Wnt signaling pathway MEC-1 cells (Nanjing Kebai Biotechnology, catalog number: CBP60514) naturally express the ROR1 ligand Wnt5a. Based on this cell, we analyzed the inhibition of the non-classical Wnt signaling pathway induced by Wnt5a stimulation by anti-ROR1 humanized antibody.

[0332] The target gene encoding human ROR1 (SEQ ID NO: 95) was transfected into 293T-NF-κB cells to obtain a stable cell line 293T-ROR1-NF-κB.

[0333] 293T-ROR1-NF-κB cells stably expressing ROR1 were adjusted to a cell density of 2 × 10⁻⁶ cells using DMEM medium containing 10% FBS. 5 Cells / mL, 100 μL / well seeded in 96-well blank plates (Costar). ®(Catalogue No.: 3917), incubated overnight. Take the 96-well blank plate, discard the supernatant, add 100 μL / well of anti-ROR1 humanized antibody diluted with MEC-1 cell culture supernatant (final in-plate concentrations of 20 nM, 10 nM, and 2.5 nM), and incubate for 24 h in a 5% CO2, 37℃ incubator; a blank control group was set up without antibody. Take the Bio-Lite Luciferase Assay System detection reagent (Vazyme... ® (Catalog No.: DD1201-03) After equilibration to room temperature, add the above-mentioned detection reagent (100 μL / well) to a 96-well white plate. After reacting for 3 min, detect the chemiluminescence signal using a multi-functional microplate reader (PE, model Envision 2105). Analyze the data, calculate the inhibition rate, and the results are presented in [the table / document name is missing]. Figure 4 The results show that the humanized antibodies against ROR1, hz29H5D8-3.2, hz135A2B10-1.1, and hz137G11D10-2.1, all inhibited the non-classical Wnt signaling pathway stimulated by Wnt5a, and were all superior to UC-961.

[0334] Inhibition rate (%) = [(RLU value of blank control group - RLU value of antibody group) / RLU value of blank control group] × 100% Example 10: ADCC effect of anti-ROR1 humanized antibody Based on the reporter gene assay, using PA-1 cells as the target cell line, the ADCC activity of recombinant Jurkat cells (BPS Bioscience™, catalog number: 60541) was detected using humanized anti-ROR1 antibodies, with isotype IgG used as a negative control. PA-1 cells were adjusted to a cell density of 2 × 10⁶ cells / cells using MEM medium containing 10% FBS. 5 Recombinant Jurkat cells were seeded at 100 μL / well in 96-well blank plates and incubated overnight. The supernatant was discarded from the 96-well blank plates, and 50 μL / well of serially diluted anti-ROR1 humanized antibody (starting at a final in-plate concentration of 13.3 nM, serially diluted 5-fold) was added to the experimental medium (RPMI-1640 medium containing 2% FBS). The recombinant Jurkat cells were then adjusted to a cell density of 4 × 10⁶ cells / well using experimental medium. 6 The assay result was 50 μL / well inoculated into the 96-well white plate and incubated for 6 h in a 5% CO2, 37°C incubator. The Bio-Lite Luciferase AssaySystem reagent (Vazyme) was then used. ®(Catalogue No.: DD1201-03) After equilibration to room temperature, add the above-mentioned detection reagent (100 μL / well) to a 96-well white plate. After reacting for 3 min, detect the chemiluminescence signal using a multi-functional microplate reader. Analyze the data and calculate EC50. 50 ADCC activity of anti-ROR1 humanized antibodies, such as Figure 5 As shown, the calculated EC 50 As shown in Table 8.

[0335] Table 8. Effects of anti-ROR1 humanized antibody on ADCC in PA-1 cells

[0336] Example 11: Preparation of the joint-load capacity MC-GGFG-Eribulin

[0337] Compound A (100 mg, 0.12 mmol) and compound B (75 mg, 0.145 mmol) were weighed and added to a 20 mL reaction tube (CAS No. 441045-17-6 for compound A and CAS No. 2413428-36-9 for compound B). 2 mL of N,N-dimethylformamide was added. The mixture was cooled to 0 °C. 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (70 mg, 0.18 mmol) and N,N-diisopropylethylamine (49 mg, 0.36 mmol) were added, and the mixture was reacted at 0 °C for 1 h. Approximately 70 mg of compound MC-GGFG-Eribulin was obtained by liquid chromatography.

[0338] ESI-MS analysis showed that MC-GGFG-Eribulin had an m / z value of 1241.72 [M+H]. + The proton spectrum is as follows: 1H NMR (500 MHz, DMSO) δ 8.22 (t, J = 5.3 Hz, 1H), 8.12 (d, J = 8.0Hz, 1H), 8.06 (t, J = 5.4 Hz, 1H), 7.99 (t, J = 5.3 Hz, 1H), 7.65 (d, J = 5.2Hz, 1H), 7.30-7.21 (m, 4H), 7.18 (d, J = 6.4 Hz, 1H), 6.99 (s, 2H), 5.02 (d,J = 26.0 Hz, 2H), 4.79 (d, J = 38.9 Hz, 2H), 4.65-4.60 (m, 2H), 4.56 (d, J =3.7 Hz, 1H), 4.50-4.42 (m, 1H), 4.26 (d, J = 10.1 Hz, 1H), 4.21-4.14 (m, 1H), 4.10 (s, 3H), 4.05-3.98 (m, 1H), 3.86-3.64 (m, 8H), 3.60-3.45 (m, 4H), 3.37-3.30 (m, 2H), 3.26 (s, 4H), 3.17-3.09 (m, 1H), 3.08-2.99 (m, 2H), 2.84 (d, J= 10.4 Hz, 1H), 2.86-2.66 (m, 3H), 2.56-2.50 (m, 1H), 2.37-2.18 (m, 5H),2.15-2.05 (m, 3H), 2.05-1.96 (m, 2H), 1.95-1.84 (m, 4H), 1.75-1.57 (m, 6H),1.55-1.38 (m, 6H), 1.35-1.25 (m, 3H), 1.23-1.12 (m, 3H), 1.03 (d, J = 6.0 Hz, 3H), 1.00-0.92 (m, 1H).

[0339] Example 12 Preparation of anti-ROR1 antibody drug conjugate Example 12-1 Preparation of anti-ROR1 antibody-drug conjugate hzAb58-2.1-Eribulin-4(1) Reagents: Antibody: Anti-ROR1 antibody hzAb58-2.1 (i.e., the above-mentioned hz137G11D10-2.1); Connector - Payload: MC-GGFG-Eribulin.

[0340] Experimental procedure: 1. Antibody reduction: The antibody was displaced into histidine buffer (pH 6.0, L-histidine 1.43 mg / mL, L-histidine hydrochloride monohydrate 2.27 mg / mL), and the antibody concentration was adjusted to 9.92 mg / mL with histidine buffer (pH 6.0). 2 mL of the antibody solution was transferred into a light-protected EP tube, and the pH was adjusted to approximately 7.0 with 0.3 M Na2HPO4. 35 μL of 10 mM TCEP·HCl (tris(2-carboxyethyl)phosphine hydrochloride) solution was added to make the antibody to TCEP·HCl molar ratio 1:2.58. The solution was incubated at 4°C in the dark for 5 hours with gentle stirring to reduce the disulfide bonds between the antibody chains, yielding reaction solution 1.

[0341] 2. Coupling of antibody and adapter-loador: Add 85 μL of adapter-loador solution with a concentration of 10 mg / mL dissolved in 50% acetone / water solution to reaction solution 1, so that the molar ratio of antibody to adapter-loador is 1:5.1; incubate at 4℃ in the dark for 40 min, stirring gently to link the antibody and adapter-loador, to obtain reaction solution 2.

[0342] 3. Conjugation termination and purification: reaction solution 2 was ultrafiltered with pH 6.0 histidine buffer to obtain the anti-ROR1 antibody-drug conjugate hzAb58-2.1-Eribulin-4(1).

[0343] Referring to the method of Example 13, the DAR value of hzAb58-2.1-Eribulin-4(1) was measured to be 4.6.

[0344] The structure of hzAb58-2.1-Eribulin-4(1) is as follows: .

[0345] Example 12-2 Preparation of anti-ROR1 antibody-drug conjugate hzAb58-2.1-Eribulin-4(2) Reagents: Antibody: Anti-ROR1 antibody hzAb58-2.1; Connector - Payload: MC-GGFG-Eribulin.

[0346] Experimental procedure: 1. Antibody Reduction: The antibody was displaced into histidine buffer (pH 6.0, L-histidine 1.43 mg / mL, L-histidine hydrochloride monohydrate 2.27 mg / mL), and the antibody concentration was adjusted to 10.049 mg / mL using histidine buffer (pH 6.0). 30 mL of the antibody solution was placed in a light-proof glass bottle, and the pH was adjusted to approximately 7.0 using 0.3 M Na2HPO4. 526 μL of 10 mM TCEP·HCl (tris(2-carboxyethyl)phosphine hydrochloride) solution was added to make the antibody to TCEP·HCl molar ratio 1:2.58. The solution was incubated at 4°C in the dark for 5 hours with gentle stirring to reduce the disulfide bonds between the antibody chains, yielding reaction solution 1.

[0347] 2. Coupling of antibody and adapter-loador: Add 1289 μL of adapter-loador solution with a concentration of 10 mg / mL dissolved in 50% acetone / water solution to reaction solution 1, so that the molar ratio of antibody to adapter-loador is 1:5.1; incubate at 4℃ in the dark for 40 min, stirring gently to link the antibody and adapter-loador, to obtain reaction solution 2.

[0348] 3. Conjugation termination and purification: reaction solution 2 was ultrafiltered with pH 6.0 histidine buffer to obtain the anti-ROR1 antibody-drug conjugate hzAb58-2.1-Eribulin-4(2).

[0349] Referring to the method of Example 13, the DAR value of hzAb58-2.1-Eribulin-4(2) was measured to be 4.1.

[0350] The structure of hzAb58-2.1-Eribulin-4(2) is as follows: .

[0351] Example 12-3 Preparation of anti-ROR1 antibody-drug conjugate hzAb19-3.2-Eribulin-4(1) Reagents: The antibody was the anti-ROR1 antibody hzAb19-3.2 (i.e., the above-mentioned hz29H5D8-3.2); the linker-loaded agent was MC-GGFG-Eribulin. The anti-ROR1 antibody-drug conjugate hzAb19-3.2-Eribulin-4(1) was prepared according to the method of Examples 12-2.

[0352] Referring to the method of Example 13, the DAR value of hzAb19-3.2-Eribulin-4(1) was measured to be 3.8.

[0353] The structure of hzAb19-3.2-Eribulin-4(1) is as follows: .

[0354] Example 12-4 Preparation of anti-ROR1 antibody-drug conjugate hzAb56-1-1.1-Eribulin-4(1) The antibody was an anti-ROR1 antibody hzAb56-1-1.1 (i.e., the above-mentioned hz135A2B10-1.1); the linker-loaded agent was MC-GGFG-Eribulin. The anti-ROR1 antibody-drug conjugate hzAb56-1-1.1-Eribulin-4(1) was prepared according to the method of Examples 12-2.

[0355] Referring to the method of Example 13, the DAR value of hzAb56-1-1.1-Eribulin-4(1) was measured to be 3.8.

[0356] The structure of hzAb56-1-1.1-Eribulin-4(1) is as follows: .

[0357] Example 12-5 Preparation of anti-ROR1 antibody-drug conjugate hzAb58-2.1-Eribulin-8(1) Reagents: Antibody: Anti-ROR1 antibody hzAb58-2.1; Connector - Payload: MC-GGFG-Eribulin.

[0358] Experimental procedure: 1. Antibody reduction: The antibody was displaced into histidine buffer (pH 6.0, L-histidine 1.43 mg / mL, L-histidine hydrochloride monohydrate 2.27 mg / mL), and the antibody concentration was adjusted to 9.92 mg / mL with histidine buffer (pH 6.0). 5 mL of this solution was transferred into a light-protected EP tube, and the pH was adjusted to approximately 7.0 with 0.3 M Na2HPO4. 285 μL of 10 mM TCEP·HCl (tris(2-carboxyethyl)phosphine hydrochloride) solution was added to make the antibody to TCEP·HCl molar ratio 1:8.5. The solution was incubated at 37°C in the dark for 1 hour with gentle stirring to reduce the disulfide bonds between the antibody chains, yielding reaction solution 1.

[0359] 2. Coupling of antibody and adapter-loador: Add 458 μL of adapter-loador solution with a concentration of 10 mg / mL dissolved in DMSO solution to reaction solution 1, so that the molar ratio of antibody to adapter-loador is 1:11; incubate at 37°C in the dark for 1 hour, stirring gently to link the antibody and adapter-loador, to obtain reaction solution 2.

[0360] 3. Conjugation termination and purification: reaction solution 2 was ultrafiltered with pH 6.0 histidine buffer to obtain the anti-ROR1 antibody-drug conjugate hzAb58-2.1-Eribulin-8(1).

[0361] Referring to the method of Example 13, the DAR value of hzAb58-2.1-Eribulin-8(1) was measured to be 7.7.

[0362] The structure of hzAb58-2.1-Eribulin-8(1) is as follows: .

[0363] Examples 12-6 Preparation of control anti-ROR1 antibody-drug conjugate MK-2140 Reagents: Antibody: UC-961; Connector-payload: MC-VC-PABC-MMAE (MCE, catalog number: HY-15575). Anti-ROR1 antibody-drug conjugate MK-2140 was prepared according to the method described in Examples 12-2.

[0364] The structure of MC-VC-PABC-MMAE is as follows:

[0365] Referring to the method in Example 13, the DAR value of MK-2140 was measured to be 4.1.

[0366] The structure of the MK-2140 is as follows: .

[0367] Example 13 Determination of DAR value of antibody-drug conjugate The DAR value was determined by hydrophobic interaction chromatography (HIC). The neutral high-salt mobile phase was used to enhance the hydrophobicity of protein molecules, thereby enabling them to bind with hydrophobic bonds in the chromatographic column. Then, the elution was carried out by gradually decreasing the salt concentration and gradually increasing the proportion of isopropanol. The less hydrophobic components were eluted first, and the more hydrophobic components were eluted later.

[0368] The components of antibody-drug conjugates (ADCs) were separated using nonporous resin (NPR) packed with bonded butyl groups. The chromatographic column specifications were TSKgel Butyl-NPR, 2.5 µm, 4.6 × 100 mm, with a column temperature of 25℃. Mobile phase A was 25 mmol / L phosphate buffer-1.5 mol / L ammonium sulfate, pH 7.0. Mobile phase B was 25 mmol / L phosphate buffer-25% isopropanol, pH 7.0. The sample was diluted 1-fold with the initial proportioning mobile phase to prepare the test solution. The injection volume was adjusted according to the sample concentration, and 50 μg of protein was injected for detection at 280 nm. The flow rate was 0.5 mL / min, and gradient elution was performed for 30 min. The gradient elution parameters are shown in Table 9. Data processing was performed, and the results were quantitatively analyzed using the area normalization method. The peak area percentage of ADCs containing 0, 1, 2, 3, 4, 5, 6, 7, and 8 cytotoxic drugs was calculated, and the DAR value was also calculated. The calculation formula is: DAR value = (percentage of ADC peak area containing 0 cytotoxic drugs × 0 + percentage of ADC peak area containing 1 cytotoxic drug × 1 + percentage of ADC peak area containing 2 cytotoxic drugs × 2 + percentage of ADC peak area containing 3 cytotoxic drugs × 3 + percentage of ADC peak area containing 4 cytotoxic drugs × 4 + percentage of ADC peak area containing 5 cytotoxic drugs × 5 + percentage of ADC peak area containing 6 cytotoxic drugs × 6 + percentage of ADC peak area containing 7 cytotoxic drugs × 7 + percentage of ADC peak area containing 8 cytotoxic drugs × 8) / 100%.

[0369] Table 9. Gradient elution parameters

[0370] Example 14: Validation of Antibody-Drug Conjugate Aggregates Gel chromatography was used to separate the components of antibody-drug conjugate samples. A neutral pH buffer with 10% isopropanol was used as the mobile phase for elution, with components eluted sequentially in descending molecular weight. The column used was an ACQUITY UPLC Protein BEH SEC Column (200 Å, 1.7 μm, 4.6 × 300 mm) at 25 °C. The mobile phase was 50 mmol / L phosphate buffer-200 mmol / L sodium chloride-10% isopropanol, pH 7.0. A precise 20 μg sample was injected into the HPLC system and detected at 280 nm. The flow rate was 0.3 mL / min, and isocratic elution was performed for 15 min. Data processing was performed, and the results were quantitatively analyzed using the area normalization method. The peak area percentages of aggregates, immunoglobulin monomers, and low molecular weight impurities were calculated, with the peak preceding the main peak representing aggregates, the main peak representing immunoglobulin monomers, and the peak following the main peak representing low molecular weight impurities.

[0371] The test results are shown in Table 10.

[0372] Table 10. SEC analysis of antibody-drug conjugates

[0373] Example 15 Affinity of Antibody-Drug Conjugates The affinity of anti-ROR1 antibody, anti-ROR1 antibody-drug conjugate, and ROR1 antigen was detected using a biomolecular interaction analysis system (GE, Biacore 8K). Amino-conjugated anti-hIgG (Fc) antibody (GE, catalog No. BR-1008-39) was fed into the CM5 sensor chip. The antibody and antibody-drug conjugate were diluted to 2 μg / mL using running buffer (437 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4·12H2O, 1.8 mM KH2PO4, 0.05% surfactant P-20 (w / v), pH 7.4) and captured through the experimental channel at a flow rate of 30 μL / min. ROR1 antigen was diluted with running buffer to 50 nM, 25 nM, 12.5 nM, 6.25 nM, 3.125 nM, and 1.5625 nM. Binding was performed at a flow rate of 50 μL / min for 200 s, followed by dissociation at a time of 300 s. Data was acquired in real-time using Biacore 8K Control Software 3.0 and analyzed using Biacore Insight Evaluation Software 3.0. The binding rate constant k was calculated using a Langmuir 1:1 model. a (1 / Ms), dissociation rate constant k d (1 / s), equilibrium dissociation constant K D (M) value.

[0374] The test results are shown in Table 11.

[0375] Table 11. Affinity of Antibody-Drug Conjugates

[0376] Example 16 Cell-binding activity of anti-ROR1 antibody-drug conjugate Flow cytometry was used to detect the cell-binding activity of the anti-ROR1 antibody-drug conjugates prepared in the examples. Cell lines with different ROR1 expression levels (NCI-N87, PA-1, NCI-H1975, A549, Jeko-1, and SW620) were adjusted to a density of 2 × 10⁻⁶ cells / year. 6Add 50 μL / well of anti-ROR1 antibody-drug conjugate to each well of a 96-well plate, along with serially diluted FACS buffer (Miltenyi Biotec, catalog number 130-091-221). For Jeko-1 cells, start at 6.67 nM (ADC) with 5-fold serial dilutions (9 concentrations total); for SW620 cells, start at 1.67 nM with 4-fold serial dilutions (6 concentrations total); for NCI-N87, PA-1, NCI-H1975, and A549 cells, start at 10 nM with 3-fold serial dilutions (10 concentrations total). Incubate at 4°C for 30 minutes, centrifuge at 1000 rpm for 5 minutes, discard the supernatant, wash three times with pre-chilled FACS buffer, and add goat anti-human IgG Fcγ-PE secondary antibody (Jackson) diluted 1:200 (v / v). Immunoresearch (catalog number 109-116-170), 100 μL / well, incubated at 4°C for 20 min. Wash three times with pre-chilled FACS buffer, resuspend in 50 μL FACS buffer, and then analyze the fluorescence signal using flow cytometry (Sartorius, iQUE3). The binding activity of the anti-ROR1 antibody-drug conjugate to ROR1 on the cell surface was represented by the mean fluorescence intensity (MFI). Data analysis was performed, and the results are as follows. Figures 6A-6F As shown, the calculated EC 50 The results are shown in Tables 12-1 and 12-2 below. The results showed that hzAb58-2.1-Eribulin-4(1), hzAb58-2.1-Eribulin-4(2) and hzAb58-2.1-Eribulin-8(1) could effectively and specifically bind to cells with different levels of ROR1 expression.

[0377] Table 12-1. Cell-binding activity of anti-ROR1 antibody-drug conjugates

[0378] Table 12-2. Cell-binding activity of anti-ROR1 antibody-drug conjugates

[0379] Example 17 Internalization of anti-ROR1 antibody-drug conjugate Flow cytometry was used to detect the internalization of the anti-ROR1 antibody-drug conjugates prepared in the examples in cells with different expression levels. The cell densities of MDA-MB-468-ROR1, NCI-N87, PA-1, NCI-H1975, A549, and Jeko-1 cells with different ROR1 expression levels were adjusted to 2 × 10⁻⁶ cells / year. 6Cells / mL, 50 μL / well for 96-well plates. Sample preparation: For MDA-MB-468-ROR1 cells, the antibody-drug conjugate (ADC) sample was pre-diluted to a concentration of 20 nM (based on ADC), labeled S1, and then serially diluted 3-fold to obtain samples S1-S9; for PA-1 and NCI-H1975 cells, the ADC sample was pre-diluted to a concentration of 20 nM, labeled S1, and then serially diluted 3-fold to obtain samples S1-S10; for Jeko-1 cells, the ADC sample was pre-diluted to a concentration of 2.22 nM, labeled S1, and then serially diluted 3-fold to obtain samples S1-S7; for NCI-N87 cells, the ADC sample was pre-diluted to a concentration of 2.22 nM, labeled S1, and then serially diluted 3-fold to obtain samples S1-S8; for A549 cells, the ADC sample was pre-diluted to a concentration of 6.67... nM, labeled S1, was serially diluted 3-fold to obtain samples S1-S9. The serially diluted sample solutions were added to cell culture plates, 50 μL per well, and incubated at 4°C for 30 min. After incubation, the 96-well cell culture plate was removed, centrifuged at 400 g for 4 min at 4°C, and the supernatant was discarded. pHrodo-labeled goat anti-human IgG antibody was diluted according to the plate distribution, mixed, and 100 μL was added to each well, then incubated at 4°C. After 30 min, the plate was washed, and 40 μL of cell culture medium was added to each well. The plate was then incubated at 37°C for 2 h for internalization. The cells were then placed in a flow cytometer (Sartorius, iQUE3) to measure the fluorescence reading of the RL1 channel. Data were analyzed.

[0380] Test results are shown Figures 7A-7F See Tables 13-1 and 13-2.

[0381] Table 13-1 Internalization of Anti-ROR1 Antibody-Drug Conjugates

[0382] Table 13-2. Internalization of anti-ROR1 antibody-drug conjugates

[0383] Example 18: Killing of tumor cells by anti-ROR1 antibody-drug conjugate To detect the killing effect of the anti-ROR1 antibody-drug conjugate prepared in the examples on ROR1-positive tumor cells, the killing activity was tested using constructed ROR1-overexpressing cells NCI-H1975-ROR1, MDA-MB-468-ROR1 and natural cells NCI-N87, PA-1 and Jeko-1.

[0384] Cells in the logarithmic growth phase were harvested, and the cell densities of NCI-H1975-ROR1, MDA-MB-468-ROR1, and NCI-N87 cells were adjusted to 5 × 10⁻⁶. 3 cells / mL, 5 × 10 4 cells / mL, 2 × 10 4 Adjust the PA-1 cell density to 1 × 10⁶ cells / mL. 4 Jeko-1 cell density was adjusted to 2 × 10⁶ cells / mL. 4The sample was added at a rate of 100 μL / well to a 96-well plate and incubated at 37°C with 5% CO2 for 4-6 h. Anti-ROR1 antibody-drug conjugates were prepared in complete medium containing 10% FBS. For NCI-H1975-ROR1 cells, hzAb58-2.1-Eribulin-4(1) and MK-2140 were started at a final concentration of 8.54 nM (based on ADC), and hzAb58-2.1-Eribulin-8(1) was started at a final concentration of 5.47 nM, with 5-fold serial dilutions for a total of 9 concentrations. For MDA-MB-468-ROR1 cells, the starting concentration was 40 nM, with 4-fold serial dilutions for a total of 9 concentrations. For PA-1 cells, hzAb58-2.1-Eribulin-4(2) was started at a final concentration of 10 nM, hzAb58-2.1-Eribulin-8(1) was started at a final concentration of 1 nM, and MK-2140 was started at a final concentration of 160 nM. For NCI-N87 cells, hzAb58-2.1-Eribulin-4(2), hzAb58-2.1-Eribulin-8(1), and MK-2140 were started at a final concentration of 13.3 nM and were serially diluted 4 times to a total of 9 concentrations. For Jeko-1 cells, hzAb58-2.1-Eribulin-4(2) and hzAb58-2.1-Eribulin-8(1) were started at a final concentration of 13.3 nM and were serially diluted 3 times to a total of 11 concentrations. MK-2140 was started at a final concentration of 120 nM and was serially diluted 3 times to a total of 10 concentrations. Cells that had adhered to the plate overnight were removed. 50 μL of diluted anti-ROR1 antibody-drug conjugate was added to each well in the experimental group, while 50 μL of complete culture medium containing 10% FBS was added to each well in the control group. Cells were cultured for 120 h, 144 h, or 168 h, and then detected using the CellTiter-GloLuminescent Cell Kit (Promega, catalog number: G7572). Specifically, 75 μL of CTG detection solution (Promega, catalog number: G7572) was added to each well of a 96-well plate, vortexed, and incubated at room temperature in the dark for 10 min. Then, 180 μL of the solution was transferred from each well to an opaque white plate, air bubbles were removed, the chemiluminescence value was read, and the cell killing rate was calculated.

[0385]

[0386] Data analysis was conducted, and the results are as follows: Figures 8A-8E As shown, the calculated EC 50The results are shown in Tables 14-1 and 14-2 below. The results show that hzAb58-2.1-Eribulin-4(1), hzAb58-2.1-Eribulin-4(2), and hzAb58-2.1-Eribulin-8(1) can effectively kill tumor cells with different ROR1 expression levels, and the killing effect is better than MK-2140.

[0387] Table 14-1. Killing effect of anti-ROR1 antibody-drug conjugates on tumor cells

[0388] Table 14-2. Killing effect of anti-ROR1 antibody-drug conjugates on tumor cells

[0389] Example 19 Bystander effect of anti-ROR1 antibody-drug conjugate on tumor cells To detect the bystander effect of ADCs, NCI-H1975-ROR1 cells were used as positive cells and Jurkat cells as negative cells, and the detection was performed using a FACS-based method. NCI-H1975-ROR1 and Jurkat cells in logarithmic growth phase were harvested and their viable cell density was adjusted to 6 × 10⁶. 4 Cells were added at a concentration of 1 mL / well to each well of a 6-well plate and incubated at 37°C with 5% CO2 for 3 h to allow adhesion. Then, the ADC was pre-diluted to 0.2 nM (final concentration) and added to each well in the order of the wells, 2 mL per well. After 4 days of incubation, cells from the 6-well plates were harvested, digested with trypsin, and counted. Cells from different wells were collected, and the cell density was adjusted to 1 × 10⁻⁶ cells / well using FACS buffer. 6 Cells were added at a rate of 100 μL / well to a 96-well V plate. Cells were incubated with a live / dead dye (BD, catalog number: 565388) at room temperature for 15 min, washed twice, and then incubated with ROR1 direct-labeled antibody (Biolegend, catalog number: 357804) at 4°C for 40 min. After incubation, cells were washed twice, resuspended in FACS buffer, and the signal was detected using a flow cytometer (Invitrogen, model: AttuneNxT).

[0390] Analyze the data; see details below. Figure 9 The results showed that hzAb58-2.1-Eribulin-4(2), hzAb58-2.1-Eribulin-8(1) and MK-2140 all exhibited the bystander effect.

[0391] Example 20: Anti-ROR1 antibody-drug conjugate in nude mouse xenograft tumor model of NCI-H1975 human lung cancer cells pharmacodynamic evaluation of type SPF-grade male nude mice (NDG, source: Changzhou Cavens Laboratory Animal Co., Ltd.) were subcutaneously injected with NCI-H1975 human non-small cell lung cancer tumor masses in the right axilla. The tumors were cultured until the average volume reached 150 mm. 3 At approximately 10:00 AM, the animals were divided into 8 groups of 5 animals each. The specific grouping and dosage are shown in Table 15. Table 15. Mouse grouping and dosage

[0392] “NA” indicates that it is not applicable.

[0393] Day 0 was designated as the day of grouping. The drug was administered once via tail vein on day 0, and then once every 4 days thereafter. Tumor volume was measured 2-3 times per week, and mouse weight was recorded. General mouse behavior was observed and recorded daily. At the end of the experiment, the tumors were removed, weighed, and photographed.

[0394] The detection indicators and calculation formulas are as follows: Tumor volume, TV (mm) 3 ) = 1 / 2 × (a × b 2 ); where a is the long diameter of the tumor and b is the short diameter of the tumor.

[0395] Relative to tumor volume, RTV = TV t / TV0; where TV0 is the tumor volume on day 0, TV t This represents the tumor volume at each measurement.

[0396] Relative tumor proliferation rate, T / C (%) = T RTV / C RTV × 100%; where T RTV For the treatment group, RTV; C RTV The solvent control group is RTV.

[0397] Tumor growth inhibition rate, TGI (%) = (1 - TW / TW0) × 100%; where TW is the tumor weight in the treatment group and TW0 is the tumor weight in the solvent control group.

[0398] Weight change rate, WCR (%) = (Wt) t -Wt0) / Wt0× 100%; where Wt0 is the mouse's body weight on day 0, Wt t The mouse's weight at each measurement.

[0399] The results of various test indicators and the changes in tumor volume over time on day 30 are shown in Table 16 and below. Figure 10As shown, in the NCI-H1975 human non-small cell lung cancer model, at a low dose of the same toxin (0.02 mpk), hzAb58-2.1-Eribulin-8(1) and hzAb58-2.1-Eribulin-4(1) were significantly more effective than MK-2140. No significant toxicity was observed in any of the treatment groups.

[0400] Table 16. Effects of anti-ROR1 antibody-drug conjugates on various parameters in the NCI-H1975 human lung cancer cell subcutaneous metastasis model in nude mice.

[0401] Example 21: Killing of tumor cells by anti-ROR1 antibody-drug conjugate To detect the killing effect of the anti-ROR1 antibody-drug conjugate prepared in the examples on ROR1-positive tumor cells, the killing activity was tested using natural PA-1 and Jeko-1 cells.

[0402] Cells in the logarithmic growth phase were harvested, and the cell density of PA-1 cells was adjusted to 1 × 10⁻⁶. 4 Jeko-1 cells were increased to a density of 2 × 10⁶ cells / mL. 4 The concentration of anti-ROR1 antibody-drug conjugates was increased to 100 μL / well in a 96-well plate and incubated at 37°C with 5% CO2 for 4 h. The anti-ROR1 antibody-drug conjugates were prepared in complete medium containing 10% FBS. For PA-1 cells, the sample was started at a final concentration of 10 nM (based on ADC) and serially diluted 4-fold to obtain 9 concentrations. For Jeko-1 cells, the sample was started at a final concentration of 13.3 nM and serially diluted 3-fold to obtain 9 concentrations. Remove the cell culture plate. Add 50 μL of diluted anti-ROR1 antibody-drug conjugate per well to the experimental group and 50 μL of complete culture medium containing 10% FBS per well to the control group. Continue culturing for 120 h and 168 h, then add CCK-8 solution (Dojin Chemicals, Japan, catalog number: CK04). Specifically, remove the 96-well plate, add 15 μL of CCK-8 stock solution to each well, incubate at 37°C for 2 h, and read the OD values ​​at 450 nm and the reference wavelength of 620 nm, and calculate the cell killing rate.

[0403]

[0404] Data analysis was conducted, and the results are as follows: Figures 11A-11B As shown, the calculated EC 50 The results are shown in Table 17. The results show that hzAb19-3.2-Eribulin-DAR4(1) and hzAb56-1-1.1-Eribulin-DAR4(1) can effectively kill tumor cells with different ROR1 expression levels.

[0405] Table 17. Killing effect of anti-ROR1 antibody-drug conjugates on tumor cells

[0406] The sequence information disclosed herein is summarized in Table S2 below.

[0407] Table S2. Sequence Information

[0408] For purposes of description and disclosure, all patents, patent applications, and other identified publications are expressly incorporated herein by reference. These publications are provided solely because their publications predate the filing date of this disclosure. All statements regarding the dates of these documents or representations of their contents are based on information available to the applicant and do not constitute any acknowledgment of the accuracy of the dates or contents of these documents. Furthermore, in any country, any reference to these publications herein does not constitute an endorsement that such publications are part of the general knowledge in the art.

[0409] Although this disclosure has been described in detail above with general descriptions and specific embodiments, modifications or improvements can be made to it, which will be obvious to those skilled in the art. Therefore, such modifications or improvements made without departing from the spirit of this disclosure are all within the scope of protection claimed by this disclosure.

Claims

1. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, wherein the antibody-drug conjugate comprises an anti-ROR1 antibody or an antigen-binding fragment thereof, wherein, The anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1 containing the amino acid sequence shown in SEQ ID NO: 1, 9, 17 or 25; HCDR2 containing the amino acid sequence shown in SEQ ID NO: 2, 10, 18, 26, 49 or 50; HCDR3 containing the amino acid sequence shown in SEQ ID NO: 3, 11, 19 or 27; LCDR1 containing the amino acid sequence shown in SEQ ID NO: 4, 12, 20 or 28; LCDR2 containing the amino acid sequence shown in SEQ ID NO: 5, 13, 21 or 29; and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 6, 14, 22 or 30.

2. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to claim 1, wherein, The anti-ROR1 antibody or its antigen-binding fragment comprises: (1) HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 1, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 2, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 3, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 4, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 5, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 6; (2) HCDR1 comprising the amino acid sequence shown in SEQ ID NO: 9, HCDR2 comprising the amino acid sequence shown in SEQ ID NO: 50, HCDR3 comprising the amino acid sequence shown in SEQ ID NO: 11, LCDR1 comprising the amino acid sequence shown in SEQ ID NO: 12, LCDR2 comprising the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 comprising the amino acid sequence shown in SEQ ID NO: 14; X1 in SEQ ID NO: 50 is selected from S or A, X2 is selected from A or Q, and X3 is selected from K or Q; (3) comprising SEQ ID NO: HCDR1 containing the amino acid sequence shown in SEQ ID NO: 10, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 11, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 12, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 14; (4) HCDR1 containing the amino acid sequence shown in SEQ ID NO: 9, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 49, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 11, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 12, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 14; (5) HCDR1 containing the amino acid sequence shown in SEQ ID NO: 17, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 18, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 19, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 10, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 10, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 11, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 12, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 13, and LCDR3 containing the amino acid sequence shown in SEQ ID NO: 14; LCDR1, which contains the amino acid sequence shown in SEQ ID NO: 21, and LCDR3, which contains the amino acid sequence shown in SEQ ID NO: 22;Or (6) HCDR1 containing the amino acid sequence shown in SEQ ID NO: 25, HCDR2 containing the amino acid sequence shown in SEQ ID NO: 26, HCDR3 containing the amino acid sequence shown in SEQ ID NO: 27, LCDR1 containing the amino acid sequence shown in SEQ ID NO: 28, LCDR2 containing the amino acid sequence shown in SEQ ID NO: 29, and LCDR3 containing the amino acid sequence shown in SEQ ID NO:

30.

3. An antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof, wherein the antibody-drug conjugate comprises an anti-ROR1 antibody or an antigen-binding fragment thereof, wherein, The anti-ROR1 antibody or its antigen-binding fragment comprises: HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 7, 15, 23, 31, 51, 52, 56, 57, 58, 60, or 61, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 8, 16, 24, 32, 53, 54, 55, 59, 62, 63, or 64.

4. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to claim 3, wherein, The anti-ROR1 antibody or its antigen-binding fragment comprises: (1) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 7, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 8; (2) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 15, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 16; (3) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 23, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 24; (4) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 31, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO:

7. (5) The amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 51, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 53; (6) The amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 51, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 54; (7) The amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 51, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 55; (8) The amino acid sequences of HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 52, and the amino acid sequences of LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO:

55. (9) The amino acid sequences of HCDR1, HCDR2 and HCDR3 in the light chain variable region shown in SEQ ID NO: 52, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 in the light chain variable region shown in SEQ ID NO: 54; (10) The amino acid sequences of HCDR1, HCDR2 and HCDR3 in the heavy chain variable region shown in SEQ ID NO: 52, and the amino acid sequences of LCDR1, LCDR2 and LCDR3 in the light chain variable region shown in SEQ ID NO: 55;(11) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 56, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 59; (12) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 57, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 59; (13) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 58, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO: 59; (14) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 60, and LCDR1, LCDR2, and LCDR3 in the light chain variable region as shown in SEQ ID NO:

60. (15) HCDR1, HCDR2, and HCDR3 in the light chain variable region as shown in SEQ ID NO: 60, and HCDR1, HCDR2, and HCDR3 in the light chain variable region as shown in SEQ ID NO: 63; (16) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 60, and HCDR1, HCDR2, and HCDR3 in the light chain variable region as shown in SEQ ID NO: 64; (17) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 61, and HCDR1, HCDR2, and HCDR3 in the light chain variable region as shown in SEQ ID NO: 62; (18) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region as shown in SEQ ID NO: 61, and HCDR1, HCDR2, and HCDR3 in the light chain variable region as shown in SEQ ID NO:

62. LCDR1, LCDR2, and LCDR3 in the light chain variable region shown in SEQ ID NO: 63; or (19) HCDR1, HCDR2, and HCDR3 in the heavy chain variable region shown in SEQ ID NO: 61, and LCDR1, LCDR2, and LCDR3 in the light chain variable region shown in SEQ ID NO:

64.

5. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of claims 1-4, wherein, The anti-ROR1 antibody or its antigen-binding fragment is a monoclonal antibody, a monospecific antibody, a multispecific antibody, a Fab fragment, an F(ab')2 fragment, an Fd fragment, an Fv fragment, a dAb, an isolated CDR, scFv, scFab, or a nanobody.

6. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of claims 1-5, wherein, The anti-ROR1 antibody or antigen-binding fragment is murine, chimeric, or humanized.

7. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, wherein, The anti-ROR1 antibody or its antigen-binding fragment comprises a heavy chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 7, 15, 23, 31, 51, 52, 56, 57, 58, 60, or 61.

8. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of claims 1-7, wherein, The anti-ROR1 antibody or its antigen-binding fragment comprises a light chain variable region having an amino acid sequence identity of at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% with the amino acid sequence shown in SEQ ID NO: 8, 16, 24, 32, 53, 54, 55, 59, 62, 63, or 64.

9. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, wherein, The anti-ROR1 antibody or its antigen-binding fragment comprises: (1) a heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 7, and a light chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 8; (2) a heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 15, and a light ... The amino acid sequence shown in SEQ ID NO: 16 has a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 23; (3) a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 24; and a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 24; (4) an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 24; The amino acid sequence shown in SEQ ID NO: 31 has a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 32;(5) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 53; (6) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 53; The amino acid sequence shown in SEQ ID NO: 54 has a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 51; (7) a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 55; and a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 55; (8) an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 55; The amino acid sequence shown in SEQ ID NO: 52 has a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 53;(9) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 52, and a light chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 54; (10) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 52, and a light ... The amino acid sequence shown in SEQ ID NO: 55 has a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 56; (11) a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 59; and a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 59; (12) an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO:

59. The amino acid sequence shown in SEQ ID NO: 57 has a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 59;(13) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 58, and a light chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 59; (14) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 60, and a light ... The amino acid sequence shown in SEQ ID NO: 62 has a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 60; (15) a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 63; and a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 63; (16) an amino acid sequence that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 63; The amino acid sequence shown in SEQ ID NO: 60 has a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 64;(17) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 62; (18) A heavy chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region having at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity with the amino acid sequence shown in SEQ ID NO: 62; The amino acid sequence shown in SEQ ID NO: 63 has a light chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 61; or (19) a heavy chain variable region that is at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 64; 10. The antibody-drug conjugate, its stereoisomer, or a pharmaceutically acceptable salt thereof according to any one of claims 1-6, wherein, The anti-ROR1 antibody or its antigen-binding fragment comprises: (1) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 7, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 8; (2) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 15, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 16; (3) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 23, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 24; (4) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 31, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 32; (5) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 53; (6) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 54; (7) a heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 53, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 54; (8) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 51, and a light chain variable region containing the amino acid sequence shown in SEQ ID NO: 55; (9) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 52, and a light chain variable region containing the amino acid sequence shown in SEQ ID NO: 53; (10) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 52, and a light chain variable region containing the amino acid sequence shown in SEQ ID NO: 54; (11) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 56, and a light chain variable region containing the amino acid sequence shown in SEQ ID NO: 59; (12) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 57, and a light chain variable region containing the amino acid sequence shown in SEQ ID NO: 59; (13) A heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 54, and a light chain variable region containing the amino acid sequence shown in SEQ ID NO: 55; (58) The heavy chain variable region of the amino acid sequence shown in SEQ ID NO: 59, and the light chain variable region containing the amino acid sequence shown in SEQ ID NO: 60; (14) The heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 60, and the light chain variable region containing the amino acid sequence shown in SEQ ID NO: 62; (15) The heavy chain variable region containing the amino acid sequence shown in SEQ ID NO: 60, and the light chain variable region containing the amino acid sequence shown in SEQ ID NO: 63.(16) A heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 60, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 64; (17) A heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 62; (18) A heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 63; or (19) A heavy chain variable region comprising the amino acid sequence shown in SEQ ID NO: 61, and a light chain variable region comprising the amino acid sequence shown in SEQ ID NO: 64.

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